ALAN DEIVID PEREIRA PRECURSORES DA DEFAUNAÇÃO NO ANTROPOCENO: FLORESTAS VAZIAS E FUNCIONALMENTE INSTÁVEIS DA MATA ATLÂNTICA SUL DO BRASIL Londrina 2020 ALAN DEIVID PEREIRA PRECURSORES DA DEFAUNAÇÃO NO ANTROPOCENO: FLORESTAS VAZIAS E FUNCIONALMENTE INSTÁVEIS DA MATA ATLÂNTICA SUL DO BRASIL Tese apresentada ao Programa de Pós-graduação em Ciências Biológicas da Universidade Estadual de Londrina - UEL, como requisito parcial para a obtenção do título de Doutor. Orientador: Prof. Dr. Mário Luís Orsi Coorientador: Dr. Juliano André Bogoni Londrina 2020 Ficha de identificação da obra elaborada pelo autor, através do Programa de Geração Automática do Sistema de Bibliotecas da UEL Pereira, Alan Deivid. Precursores da defaunação no antropoceno: Florestas vazias e funcionalmente instáveis da Mata Atlântica sul do Brasil / Alan Deivid Pereira. - Londrina, 2020. 146 f. : il. Orientador: Mário Luís Orsi. Coorientador: Juliano André Bogoni. Tese (Doutorado em Ciências Biológicas) - Universidade Estadual de Londrina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Ciências Biológicas, 2020. Inclui bibliografia. 1. Ecologia - Tese. 2. Mamíferos - Tese. 3. Biologia da Conservação - Tese. 4. Espécies Invasoras - Tese. I. Orsi, Mário Luís . II. Bogoni, Juliano André. III. Universidade Estadual de Londrina. Centro de Ciências Biológicas. Programa de Pós-Graduação em Ciências Biológicas. IV. Título. CDU 574 Powered by TCPDF (www.tcpdf.org) http://www.tcpdf.org ALAN DEIVID PEREIRA PRECURSORES DA DEFAUNAÇÃO NO ANTROPOCENO: FLORESTAS VAZIAS E FUNCIONALMENTE INSTÁVEIS DA MATA ATLÂNTICA SUL DO BRASIL Tese apresentada ao Programa de Pós-graduação em Ciências Biológicas da Universidade Estadual de Londrina - UEL, como requisito parcial para a obtenção do título de Doutor. BANCA EXAMINADORA __________________________________________ Orientador: Prof. Dr. Mário Luís Orsi Universidade Estadual de Londrina – UEL __________________________________________ Prof. Dr. Luciano Martins Verdade Universidade de São Paulo – USP __________________________________________ Prof. Dr. Vlamir José Rocha Universidade Estadual de Londrina – UEL __________________________________________ Profa. Dra. Margareth Lumy Sekiama Universidade Estadual de Londrina – UEL __________________________________________ Prof. Dr. Lucas Ribeiro Jarduli Universidade Estadual de Londrina – UEL Londrina, 09 de abril de 2020 3 Dedico esta tese a minha mãe Ana Cioczek Pereira 4 AGRADECIMENTOS Aqui se encerra um ciclo de nove anos interruptos de estudos e amor a biologia, destes três dedicados ao doutorado. Nestes anos de formação, tive o privilégio de nunca estar sozinho em minha jornada e fui agraciado pela companhia de pessoas maravilhosas em diversos aspectos. Neste sentido, penso que o ato de agradecer é ter ciência de que não conquistei nada sozinho e sou feliz por isso, assim, parafraseando o poeta americano Henry Thoreau “A felicidade só é real quando é compartilhada”, e sou grato por compartilhar tantos momentos com todos vocês. Minha lista de agradecimento é longa e peço desculpas antecipadamente aos esquecidos aqui, saibam que todos vocês tiveram um papel especial em minha jornada para me tornar um pesquisador e acima de tudo uma pessoa melhor. Agradeço primeiramente a minha família. Minha querida mãe Ana Ciozeck Pereira, por assumir o papel de mãe e pai devido ao falecimento deste. Neste papel sempre será minha primeira orientadora e maior incentivadora de meus estudos, certamente este doutorado é uma homenagem a toda sua dedicação e apoio. Agradeço aos meus irmãos Alex e Patrik por me fornecerem toda a segurança e aporte emocional que mesmo distante de nossa família, vocês se encarregaram de manter nossos laços afetivos e contribuem para o bem-estar de todos. Agradeço também a minha namorada Jessica Hainosz, que sempre esteve ao meu lado nos momentos mais difíceis me fornecendo apoio em todas as minhas decisões, certamente você foi e é meu porto seguro nesta jornada chamada vida. Agradeço imensamente ao meu orientador e amigo professor Dr. Mário Luís Orsi, por ter me aceitado como seu aluno, mesmo sendo um mastozoólogo “invasor” em seu Laboratório de Ecologia de Peixes e Invasões Biológicas (LEPIB). Certamente você é uma das maiores inspirações de profissional que eu tive o prazer de conhecer, espero seguir minha carreira com a mesma paixão, convicção e garra que o senhor demonstra todos os dias. Também sou grato ao meu segundo orientador, Dr. Juliano André Bogoni, por aceitar me orientar e sanar horas de dúvidas, discussões sobre estatísticas e algoritmos inteligentes, seus trabalhos me fazem sentir orgulho e me incentivam a continuar na pesquisa tendo mamíferos como foco de estudo, e sua humildade me fazem crer que posso chegar ainda mais longe. Agradeço a professora Dr. Ana Paula Vidotto- Magnoni por me apoiar, incentivar e inspirar a seguir com pesquisas com mamíferos na região norte do Paraná e pela parceria em diversas publicações científicas. Não menos importante agradeço ao professor Dr. Sérgio 5 Bazilio, por ser meu principal amigo e parceiro de longa data nas coletas e informações sobre a mastofauna paranaense, meu muito obrigado. Aos amigos do Laboratório de Ecologia de Peixes e Invasões Biológicas (LEPIB), Harry, Armando, Lucas, Marcelo, João, Carol (mãe), Iago, Tuco, Felipinho (Victor), Matheus, Sarah, Carol (freakinha) e Gabi. Aos amigos de outros laboratórios Carol Blefari, Ricardo, Gabriel, Tati, Guilherme (sapos), Guilherme (aves), Larissa e Gisele. Por fim sou imensamente agradecido aos técnicos da UEL Aparecido de Souza, Edson Santana da Silva e Jurandir por sempre me auxiliarem em etapas de campo. Agradeço a todos meus amigos de fora da academia em especial aos amigos Marcos e Crislaine por estarem ao meu lado a anos, sempre me incentivando a nunca parar com minhas pesquisas. Ao Programa de Pós Graduação em Ciências Biológicas da Universidade Estadual de Londrina, por todo o suporte oferecido de modo a possibilitar minha formação profissional. Ao corpo docente de professores da UEL e outras instituições por compartilharem todo seu conhecimento ao longo destes cinco anos de mestrado e doutorado, possibilitando o meu crescimento intelectual bem como a desenvolvimento do presente estudo. Ao Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio) pela autorização de pesquisa em unidadees de conservação e ao Instituto Ambiental do Paraná (IAP) pela autorização de pesquisas nos demais fragmentos. O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 1689817. 6 Não devemos deixar que uma floresta cheia de árvores nos engane, acreditando que tudo está bem. Muitas dessas florestas estão "mortas-vivas" e, embora os satélites que passam por cima delas possam registrá-las tranquilamente como floresta, elas estão vazias de grande parte da riqueza da fauna valorizada pelos seres humanos. Uma floresta vazia é uma floresta condenada. Kent H. Redford (1992) 7 Pereira, Alan Deivid. Precursores da defaunação no antropoceno: florestas vazias e funcionalmente instáveis da Mata Atlântica Sul do Brasil. 2020. 144 f. Tese (Doutorado em Ciências Biológicas) – Universidade Estadual de Londrina, Londrina, 2020. RESUMO As lacunas de informações básicas sobre a distribuição de espécies bem como a ausência de dados de abundância em séries históricas dificultam a correta avaliação do estado de defaunação em comunidades de mamíferos, nas diferentes regiões do Brasil, como é o caso da porção Sul da Mata Atlântica brasileira. Em escalas locais, acredita-se que os principais precursores associados a defaunação são a caça, introdução de espécies não nativas e conversão do habitat. Considerando os dois últimos fatores citados o presente estudo teve como objetivos investigar os mecanismos relacionados ao declínio de mamíferos de médio e grande porte em remanescentes de Mata Atlântica do Sul do Brasil. Primeiramente realizamos o primeiro inventario da mastofauna de médio e grande porte do Parque Nacional dos Campos Gerais afim de entender como as espécies ocupam esta importante unidade de conservação. Com um esforço amostral de 780 horas de busca ativa e 157.516 horas em armadilhas fotográficas, foram registradas 31 espécies de 17 famílias e 8 ordens, sendo que destas espécies, 42% são ameaçadas de extinção em alguma esfera (estadual, nacional ou internacional). No segundo capítulo investigamos o impacto de uma rodovia de pequeno porte sobre a fauna de mamíferos da região norte do estado do Paraná. Monitoramos a PR-455 durante um ano, totalizando 7.296 km percorridos em 96 campanhas. Registramos 60 indivíduos atropelados pertencentes a 17 espécies, representando uma taxa de 0,151 indivíduo / km / dia. Nossos resultados indicaram que as regiões com maior incidência de atropelamentos no PR-445 são aquelas próximas a trechos sobre rios e com remanescentes de vegetação nativa. No terceiro capítulo, usamos modelos de nicho ambiental para entender a distribuição geográfica de Myocastor coypus com base em preditores bioclimáticos, identificando áreas com maior aptidão para a invasão dessa espécie. Observamos que as áreas de maior adequabilidade climática sendo que os maiores scores da avaliação de risco de invasão estão restritas às regiões sudeste e sul do Brasil, e ainda as modificações antropogênicas da paisagem são variáveis que melhor explicam a adequabilidade e ocorrência dessa espécie em novos ambientes. No último capítulo avaliamos efeitos da conversão do habitat no declínio de mamíferos de médio e grande porte em remanescentes de Mata Atlântica, através do índice de defaunação e modelos lineares generalizados. Encontramos um alto grau de defaunação em todos fragmentos analisados com mais de 50% de todos os fragmentos analisados apresentando valores médios de defaunação superiores à média histórica esperada dos últimos 500 anos. Áreas com maior porcentagem de cobertura de solo destinadas a silvicultura e agricultura apresentam maiores valores de defaunação se comparadas com áreas com maior cobertura de florestas naturais. Concluímos que para a porção Sul da Mata Atlântica não há apenas um fator determinante de defaunação e sim um conjunto de fatores que exercem pressões cotidianas e constantes sobre as assembleias de mamíferos locais, de maneira que se medidas preventivas não forem adotadas e implementadas em curto prazo os fragmentos restantes passaram por severos processos de homogeneização de fauna. Palavras-chave: Fragmentação; Floresta Tropical; Mamíferos; Perda da biodiversidade; Uso da terra 8 Pereira, Alan Deivid. Precursors of defaunation in the Anthropocene: empty and functionally unstable forests in the South Atlantic Rainforest of Brazil. 2020. 144 pp. Thesis (Doctorate in Biological Sciences) – Universidade Estadual de Londrina, Londrina, 2020. ABSTRACT The gaps of basic information on the distribution of species as well as the absence of abundance data in historical series make it difficult to estimate the state of defaunation in many regions of Brazil, such as the southern portion of the Brazilian Atlantic Rainforest. At local scales, the main precursors associated with defaunation are believed to be hunting, the introduction of non- native species, and habitat conversion. Considering the last two factors mentioned, the present study aimed to investigate the mechanisms related to the decline of medium and large mammals in remnants of the southern Brazilian Atlantic Rainforest. Firstly, we carried out the first inventory of medium-sized and large mammals for Parque Nacional dos Campos Gerais, Paraná, Brazil to understand how species occupy this important conservation unit. With a 780- hour sampling effort in active research and 157,516 hours in camera traps, 31 species of 17 families and 8 orders were recorded. Of these species, 42% are threatened with extinction at state, national, or international level. In the second chapter, we investigate the impact of a small highway on the fauna of mammals in the northern region of the state of Paraná. We monitored the PR-455 for one year, totaling 7,296 km traveled in 96 trips. We recorded 60 roadkill mammals belonging to 17 species, representing a rate of 0.151 individual/km/day. Our results indicate that the regions with the highest incidence of roadkills on PR-445 are those close to stretches over rivers and with remnants of native vegetation. In the third chapter, we use environmental niche models to understand the geographic distribution of M. coypus based on bioclimatic predictors and to identify areas with greater suitability for the invasion of this species. We observed that the areas of greatest climatic suitability and with the highest scores for the risk of invasion assessment are restricted to the southeastern and southern regions of Brazil, and yet the anthropogenic changes in the landscape are variables that best explain the suitability and occurrence of this species in new environments. In the last chapters, we evaluated the effects of habitat conversion to the decline of medium-sized mammals in remnants of the southern Brazilian Atlantic Rainforest, through the defaunation index and generalized linear models. We found a high degree of defaunation in all analyzed fragments, with more than 50% of all analyzed fragments showing rates defaunation values higher than the expected historical average of the last 500 years. Areas with a higher percentage of soil cover for silviculture and agriculture have greater defaunation rates when compared to areas with natural forest cover. We conclude that for the South portion of the Brazilian Atlantic Rainforest there is not only one factor that determines defaunation, but a set of factors that exert daily and constant pressure on the assemblages of local mammals, so that if preventive measures are not adopted and implemented in In the short term, the remaining fragments went through severe fauna homogenization processes. Key words: Fragmentation; Land use; Loss of biodiversity; Mammals; Tropical forest 9 LISTA DE FIGURAS CAPÍTULO 1 Figura 1 – Map of the Parque Nacional dos Campos Gerais, state of Paraná, Brazil, with monitoring sites by camera traps during the 2013–2014 and 2015–2017 sampling periods............................................................................................... ... 25 Figura 2 – Figures 2–7. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 2. Didelphis albiventris. 3. Didelphis aurita. 4. Myrmecophaga tridactyla. 5. Tamandua tetradactyla. 6. Dasypus novemcinctus. 7. Euphrsctus sexcintus. ........................................................................................................... .27 Figura 3 – Figures 8–13. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 8. Mazama gouazoubira. 9. Mazama americana. 10. Mazama nana. 11. Pecari tajacu. 12. Sus scrofa .13. Sapajus nigritus. .......................................... .28 Figura 4 – Figures 14–19. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 14. Alouatta guariba clamitans.15. Cerdocyon thous. 16. Canis lupus familiaris. 17. Leopardus pardalis. 18. Leopardus guttulus. ........ .29 Figura 5 – Figures 20–25. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 20. Puma concolor. 21. Puma yagouaroundi. 22. Eira barbara. 23. Galictis cuja. 24. Nasua nasua. 25. Procyon cancrivorus.........................................................30 Figura 6 – Figures 26–30. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 26. Lepus europaeus. 27. Cuniculus paca. 28. Dasyprocta azarae. 29. Coendou spinosus. 30. Guerlinguetus brasiliensis. ............................................31 Figura 7 – The rarefaction curve (observed and estimates by jackknife 1) of mammal species registered by identifcation of footprints and trap cameras. Vertical bars represent standard deviation. CT: Camera trap; F: footprints. ....................33 CAPÍTULO 2 Figura 1 – Location of Highway PR-445 in the state of Paraná. Brazil indicating the sector measured in this study from 2018 to 2019. The hotspots highlighted in red indicate 10 places with high rates of mammal roadkill...........................................................46 Figura 2 – Figure 2. Microstructural pattern of species of the family Felidae: Leopardus wiedii. (A) cuticular pattern and (B) medullar pattern; Leopardus guttulus; (C) cuticular pattern and (D) medullar pattern; and Puma yagouaroundi (E) cuticular pattern and (F) medullar pattern..........................................................................50 Figura 3 – Boxplot illustrating the difference in roadkill rate for medium- to large- bodied mammals on Highway PR-445 between the municipalities of Londrina and Mauá da Serra, PR in relation to: (A) activity pattern; and (B) feeding habit. Acronyms are described in Table 1. A: a significant difference [p= 0.05] is shown between carnivores (CA) and frugivores/omnivores (Fr/On). B: a significant difference [p = 0.04] is shown between carnivores (CA) and insectivores/omnivores (In/On). ..........................................................51 CAPÍTULO 3 Figura 1 – Myocastor coypus records in Brazilian biomes and the suitability values for each record. .........................................................................................................67 Figura 2 – Suitability map for Myocastor coypus occurrences in Brazil (a). South and Southeast Brazil, with confirmed occurrences in the published scientific literature (b). Low suitability values are represented by green and high suitability values are represented by red.Highlighted, aphotographic record ofM. coypus through opportunistic observationsinnorthern ofthe stateof Santa Catarina and the record of a run-over specimen on the PR-538 highway, near the Cafezal River, in the municipality of Londrina, PR. .............68 Figura 3 – Surveillance map, combined binary map for the occurrence of Myocastor coypus in Brazil. (a) The southeastern and southern regions of Brazil are highlighted. (b) Record percentages in relation to risk assessment for Brazil areas .....................................................................................................................69 Support Material Figura S1 – Occurrence points before and after spatial filter. Blue bar = Raw GBIF coordinate data points before spatial filter, Purple = Thinned GBIF coordinate data points after spatial filter, Red = Raw Brazil coordinate data points before spatial filter, Green = Thinned Brazil coordinate data points before spatial filter ..............................................................................................76 Figura S2 – Ordination plot of climate variables. All results were obtained only with 11 native data according with IUCN. .......................................................................80 Figura S3 – Ordination plot of climate variables. Loading values results with native and Brazil occurrences. ..............................................................................................81 Figura S4 – Suitability models calibrated with GBIF native occurrences and projected within Brazilian range. Color gradient shows suitability values from blue (i.e., low suitability) to yellow (i.e., Highest values). .........................................83 Figura S5 – Suitability models calibrated with Brazilian occurrences (i.e., data set two) and projected within Brazilian range. Color gradient shows suitability values from blue (i.e., low suitability) to yellow (i.e., Highest values). .............84 Figura S6 – Binary classification maps for the four different niche models used to model M. coypus. Each classification maps were obtained from GBIF occurrences in native range and projected within Brazilian range. Binary models use threshold values (i.e., here sensitivity = specificity) to classify grid cells in presence and absence class. Grey color indicate cell predicted as absence and red indicate cell predicted as presence. ................................................... .....85 Figura S7 – Binary classification maps for the four different niche models used to model M.coypus. Each classification maps were obtained from Brazilian occurrences and projected within Brazilian range. Binary models use threshold values (i.e., here sensitivity = specificity) to classify grid cells in presence and absence class. Grey color indicate cell predicted as absence and red indicate cell predicted as presence .................................................... .....86 Figura S8 – Pairwise correlation values of suitability values obtained by the ensemble approach and the environmental variables used in the modeling procedures for Myocastor coypus in Brazil. ..................................................................... .....87 CAPÍTULO 4 Figura 1 – Map of the areas analyzed of the Atlantic Rainforest in southern Brazil, with special reference to EPA of the Devonian Escarpment. PCG: Parque Nacional dos Campos Gerais; RBA: Reserva Biológica das Araucárias; RPC: FNI: Floresta Nacional de Irati; RPPN Corredor do Iguaçu; FSN: Fragmento São Nicolau; FPL: Fragmento Portão lajeado; FMS: Fragmento Monte Seleto; FCA: Fragmento Caxambu; FMI: Fragmento Mirante; FBM: Fragmento Barra Mansa; FGR: Fragmento da Gruta ..........................................95 Figura 2 – Defaunation index for medium- and large-bodied mammals on study sites 12 in the southern portion of the Brazilian Atlantic Forest biome: (a) all medium- and large-bodied mammal species; (b) high trophic level; (c) intermediate tropic level; (d) low trophic level; (e) frugivores; (f) large grazers; (g) mesocarnivores; (h) apex-predators; (i) small-bodied; (j) large- bodied; (k) boxplot illustrating the difference in defaunation index between non-protected and protected areas .......................................................................98 Support Material Figure S3 – Land use cover found in forest remnants of the South Atlantic Forest in Brazil...108 13 LISTA DE TABELAS CAPÍTULO 1 Tabela 1 – List of land medium-sized and large mammal species at the Parque Nacional dos Campos Gerais, Brazil. Recordmethod; Footprint (F), Visualization (VI), Camera traps (CT), Vocalization (VO). Conservation status by (IUCN), Brazilian List of Threatened Species (BR), Paraná state List of Threatened Species (PR). Data Deficient (DD), Endangered (EN), Least Concern (LC), Not evaluated (NE), Near Threatened (NT), Vulnerable (VU) and Critically Endangered (CR). Species added in 2016 to 2017 (+). Exotic species (*). Small species (**). ................................................................................................................................................ .32 CAPÍTULO 2 Tabela 1 – List of roadkill medium- to large-bodied mammalian species on Highway PR-445. Acronyms are as follows. N: Total number of occurrences. F: Frequency of occurrence by species. Diet: carnivore (Ca); frugivore and folivore (Fr/Fo); frugivore and omnivore (Fr/On); herbivore or grazer (Hb); insectivore and omnivore (In/On) and myrmecophage (Myr). Activity pattern: crepuscular/nocturnal (CrN); diurnal (Dn); nocturnal (Nt). Threat category: data deficient (DD); endangered (EN); least concern (LC); not evaluated (NE); near threatened (NT) and vulnerable (VU). Exotic species (*). Smallbodied species (**). ..................................... .49 CAPÍTULO 3 Tabela 1 – Partial area under the curve (pAUC)and threshold values (sensitivity = specificity) for the four algorithms used to model the potential distribution of Myocastor coypus (models were combined if p AUC >1). The relative importance of each variable used to model the distribution of Myocastor coypus are displayed below the table. ................................................68 SUPPORT MATERIAL Tabela S1 – Myocastor coypus coordinates occurrence records in Brazil. Locality: Municipality in which each species coordinate was observed, State: State in which each species coordinate was observed: Rio Grande do Sul (RS); Santa Catarina (SC); Paraná (PR); São Paulo (SP); Rio de Janeiro (RJ), Biome: landscape classification in which each species coordinate was observed, Reference: information source in which each species coordinate was gathered, Map ID: point label for each occurrence in map reference (see Fig. 2 in manuscript), NR indicate ad libitum observations by S. Bazilio or M. L. Orsi and DR indicate dead species founded in the road .Suitability: suitability values for each occurrence points in Brazil. ........77 Tabela S2 – PCA loading values of GBIF thinned data set inside native range. In bold, selected bioclimatic 14 variables names and values of retained variables. PC1 = first component of PCA, PC2 = second componente of PCA. ...........................................................................................................................78 Tabela S3 – PCA loading values from Brazilian thinned data set. In bold, loading bioclimatic variables names and values of retained variables. PC1 = first component of PCA, PC2 = second component of PCA. ................................................................................................................ .79 CAPÍTULO 4 Tabela 1 – Description of sampling effort employed on all study areas in the Atlantic Rainforest, Brazil......................................................................................................................................................96 Tabela 2 – Results of GLM with a quasi-Poisson distribution of the best model for defaunation according to the covered-land use variables......................................................................................99 SUPPORT MATERIAL Tabela S1 – Landscape characterization of the analyzed areas. Areas: PCG: Parque Nacional dos Campos Gerais; RBA: Reversa Biológica das Araucárias; RPC: FNI: Floresta Nacional de Irati; RPPN Corredor do Iguaçu; FSN: Fragmento São Nicolau; FPL: Fragmento Portão lajeado; FMS: Fragmento Monte Seleto; FCA: Fragmento Caxambu; FMI: Fragmento Mirante; FBM: Fragmento Barra Mansa; FGR: Fragmento da Gruta; HB: Historical baseline................107 Tabela S2 – List of medium-sized and large mammal species in the 11 areas analyzed along with the IUCN historical baseline. Trophic Guild: Ca: Carnivore; Fr: Frugivore; Fo: Folivore; Gr: Gumivore; Hb: Herbivore grazer; In: Insectivore; Myr: Myrmecophage; On: Omnivore; Os: Piscivore. Areas: PCG: Parque Nacional dos Campos Gerais; RBA: Reversa Biológica das Araucárias; RPC: FNI: Floresta Nacional de Irati; RPPN Corredor do Iguaçu; FSN: Fragmento São Nicolau; FPL: Fragmento Portão lajeado; FMS: Fragmento Monte Seleto; FCA: Fragmento Caxambu; FMI: Fragmento Mirante; FBM: Fragmento Barra Mansa; FGR: Fragmento da Gruta; HB: Historical baseline..........................................................................................................112 Tabela S3 – Defaunation values found in forest remnants of the South Atlantic Forest in Brazil....................................................................................................................................................115 15 SUMÁRIO INTRODUÇÃO GERAL ..................................................................................................... .17 REFERÊNCIAS ................................................................................................................... .20 CAPÍTULO 1. Checklist of medium-sized to large mammals of Campos Gerais National Park, Paraná, Brazil................................................................................................................23 Abstract....................................................................................................................................24 Introduction.............................................................................................................................24 Methods....................................................................................................................................25 Study area..................................................................................................................................25 Sampling................................................................................................................................... 26 Data analysis..............................................................................................................................26 Results...................................................................................................................................... 26 Annotated list............................................................................................................................ 27 Discussion.................................................................................................................................37 Acknowledgements..................................................................................................................38 References................................................................................................................................38 CAPÍTULO 2. Narrow, short and deadly: Mammals roadkills on highway sections of PR- 445, South of Brazil…………..................................................................................................40 Abstract...………………………………………………………………………………….....42 Introduction……….................................................................................................................43 Material and methods…………………………………………………………………….....44 Study area..…………………………………………………………………………………....44 Sampling.……………………………………………………………………………………..45 Data analysis.............................................................................................................................47 Results………………………………………………………………………………………..48 Discussion………………………………………………………………………………….....51 Acknowledgments…………………………………………………………………………...55 References................................................................................................................................55 CAPÍTULO 3. Modeling the geographic distribution of Myocastor coypus (Mammalia, Rodentia) in Brazil: establishing priority areas for monitoring and an alert about the risk of invasion................................................................................................................................62 Abstract....................................................................................................................................64 Introduction………………………………………………………………………………...64 16 Material and methods...……………………………………………………………………...65 Native occurrence data………………………………………………………………………….……..65 Brazilian occurrence data……………………………………………………………………….….....65 Environmental data and variable selection……………………………………………………..…...66 Environmental niche models (ENM)………………………………………………………………...66 Model calibration and model evaluation……….…………………………………………………...66 Surveillance map…………………………….……………………………………………………........66 Results………………………………………………………………………………………..67 Discussion.................................................................................................................................69 Acknowledgments…………………………………………………………………………..71 Disclosure statement.……………………………………………………………………......71 Funding…………………………………………………………….………………………...71 Orcid.........................................................................................................................................71 References................................................................................................................................71 Support material………………………………………………………………………….....74 CAPÍTULO 4. Mammalian defaunation in Devonian kniferidges and meridional plateaus of the Brazilian Atlantic Rainforest........................................................................................91 Abstract....................................................................................................................................92 Introduction………………………………………………………………………………...93 Material and methods...………………………………………………………………………94 Study area..................................................................................................................................94 Mammal sampling and functional groups.................................................................................95 Landscape characterization.......................................................................................................97 Data analysis……………………………………………………………………………….....97 Results......................................................................................................................................98 Discussion……………….........................................................................................................99 Acknowledgments…….........................................................................................................101 References…………………………………………………………………………………..101 Support material....................................................................................................................106 CONCLUSÃO GERAL........................................................................................................116 ANEXO A: Normas técnicas - revista Papéis Avulsos de Zoologia ....................................118 ANEXO B. Normas técnicas - revista Biological Conservation.…………...........................126 17 INTRODUÇÃO GERAL Na biologia da conservação, o termo defaunação é utilizado como sinônimo da perda de espécies e populações de animais silvestres, bem como o declínio local da abundância de indivíduos (DIRZO et al., 2014). Apesar do termo já constar a mais de 25 anos no âmbito da conservação (ver. DIRZO e MIRANDA, 1990), a defaunação continua sendo um fenômeno amplamente pouco explorado (DIRZO et al., 2014) e entender a escala e as consequências da defaunação é uma prioridade crescente para ecologistas, gestores da vida selvagem e biólogos da conservação (YOUNG et al., 2016). Os mecanismos que desencadeiam eventos de defaunação atuam em escala global (e.g., mudanças climáticas e poluição de solo e água) e escalas locais, que incluem a caça comercial, introdução de espécies não nativas e conversão do habitat (DIRZO et al., 2014; YOUNG et al., 2016). Contudo, ainda não somos capazes de estabelecer um conjunto consistente de preditores universais de defaunação, particularmente em escalas locais (YOUNG et al., 2016). De maneira geral a defaunação pode afetar desproporcionalmente espécies de maior porte e baixa taxa reprodutiva, por exemplo, os grandes mamíferos (CARDILLO et al., 2005). Mamíferos de médio e grande porte desempenham papeis importantes para a manutenção e equilíbrio dos ecossistemas florestais, mediante vários serviços ecológicos realizados por este grupo (STONER et al., 2007; MARKL et al., 2012). Isso inclui controle populacional de presas, polinização de plantas e dispersão de sementes, contribuindo para a regeneração das florestas (TERBORGH et al. 1999; GALETTI et al., 2015; DERHÉ et al., 2018). Sendo assim as consequências ecológicas de sua defaunação pode interferir em diferentes escalas ecológicas promovendo efeitos em cascata na abundância, composição e ecologia de outras espécies da fauna e flora (KURTEN, 2013; DIRZO et al., 2014). Nas florestas tropicais as populações de mamíferos apresentam as maiores taxas de declínio em relação a outros lugares do mundo (DIRZO et al., 2014; BOGONI et al., 2018), sendo possível observar eventos crônicos e repetidos de defaunação (i.e., defaunações locais). Como mencionado anteriormente, em escalas locais a defaunação é principalmente relacionada a perda e conversão do 18 habitat decorrentes da urbanização, mudanças no uso da terra para fins agrícolas, desenvolvimento de estradas, exploração de madeira, mineração e caça furtiva (DIRZO et al., 2014; YOUNG et al., 2016). No Brasil, a Mata Atlântica é considerada um dos biomas mais afetados pelos processos de fragmentação e conversão de uso de terra (RIBEIRO et al., 2009). Devido à fragmentação e drástica redução da cobertura florestal que ocorreram neste bioma, as assembleias de mamíferos residem atualmente em remanescentes de florestas nativas tipicamente menores que 100 hectares e imersos em matrizes antropogênicas (RIBEIRO et al., 2009). Estima-se que 96% de todo o bioma da Mata Atlântica esteja sujeito a alguns efeitos de cascatas tróficas devido à defaunação de mamíferos (JORGE et al., 2013).Os principais impulsionadores da defaunação em toda a Mata Atlântica incluem uma longa e repetida história de pressão de caça, conversão e fragmentação de habitats, ou a combinação sinérgica de ambos (BOGONI et al., 2018). O aumento da malha viária brasileira do último século é outro fator que deve ser considerado como causa direta para o declínio de populações de mamíferos terrestres em diversos biomas. As estradas são responsáveis por diversas mudanças ambientais, resultando em um grande impacto nas paisagens naturais (LAURANCE et al., 2014). O atropelamento é o principal impacto negativo das estradas na vida selvagem, com efeitos diretos nas populações locais, influenciando a abundância e distribuição das espécies (EIGENBROD et al., 2008). Os declínios populacionais causados pelos impactos das estradas têm efeitos sobre a variabilidade e viabilidade genética das espécies (JACKSON e FAHRIG, 2011). Estima-se que 5 milhões de animais de médio e grande porte são mortos anualmente nas estradas e rodovias do Brasil (CBEE, 2019), provando este ser um fator direto de mortalidade entre os vertebrados terrestres tão significativos quanto a caça (SEILER e HELDIN, 2006). O impacto decorrente a introdução de espécies não nativas é outro fator citado com frequência como precursor do declínio de populações de fauna silvestre. Espécies invasoras representam uma das ameaças mais significativas à biodiversidade global e à função dos ecossistemas (CLOUT e RUSSELL 2007; YOUNG et al., 2016). De todos os animais extintos para 19 os quais a causa da extinção foi determinada, 54% incluíram efeitos de espécies invasoras (CLAVERO e GARCÍA-BERTHOU, 2005). As espécies exóticas invasoras comprometem as interações ecológicas nos ecossistemas, afetando negativamente as espécies nativas através da competição direta por recursos (GALETTI et al., 2015), indireta ou aparente (mediada por parasitas e patógenos) (LONG, 2003) predação (DA ROSA et al., 2017), modificação no habitat e alterações dos ciclos de nutrientes nas águas (LONG, 2003; CLOUT e RUSSELL 2007). Na Mata Atlântica brasileira, estudos em escala regional demostraram elevados níveis de defaunação em assembleias de mamíferos (GALETTI et al., 2006; CANALLE et al., 2012; BOGONI et al., 2016; GALETTI et al., 2017). A maioria dos estudos que avaliaram a defaunação na Mata Atlântica é concentrada nas regiões Centro-Oeste, Sudeste e Nordeste do Brasil (GALETTI et al., 2006, 2017; CANALE et al., 2012). Portanto, para a porção sul deste bioma ainda são escassas informações sobre os efeitos da conversão de habitats no declínio de mamíferos de médio e grande porte. O presente estudo buscou investigar mecanismos relacionados ao declínio de mamíferos em remanescentes de Mata Atlântica do Sul do Brasil. Consideramos os precursores relacionados a defaunação em escala local (e.g., introdução de espécies não nativas e conversão do habitat). Neste sentido, este estudo está divido em quatro capítulos. Capítulo 1: inicialmente apresentamos o inventário da mastofauna de médio e grande porte do Parque Nacional dos Campos Gerais, PR. Consideramos que conhecimento sobre a diversidade e a distribuição das espécies é essencial para os estudos de ecologia e conservação (OLIVEIRA et al., 2017), sendo esta a primeira ferramenta necessária para a mensuração da defaunação em escala local. Capítulo 2: nesta sessão avaliamos o impacto de uma rodovia de pequeno porte (PR-445) sobre a mastofauna de uma região de Mata Atlântica considerada densamente fragmentada. Considerando os declínios populacionais causados por atropelamentos em estradas têm efeitos tão significativos quanto a caça (SEILER e HELLDIN, 2006). Capítulo 3: Sabendo que um dos fatores relacionados a eventos de defaunação é a introdução/estabelecimento de espécies não nativas em novas áreas, neste capítulo o objetivo foi entender a distribuição geográfica de Myocastor coypus (ratão-do-banhado), e indicar áreas de maior 20 risco de estabelecimento, baseadas em preditores bioclimáticos e um mapa de vigilância. Capítulo 4: Por fim o último capítulo avaliamos efeitos da conversão do habitat em relação ao declínio de mamíferos de médio e grande porte da Mata Atlântica Sul, através do índice de defaunação. REFERÊNCIAS BOGONI, J. A. et al. 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Annual Review of Ecology, Evolution, and Systematics, v. 47, n. 1, p. 333–358, 2016. 23 CAPÍTULO 1 Checklist of medium-sized to large mammals of Campos Gerais National Park, Paraná, Brazil Alan Deivid Pereira, Sergio Bazilio, Mário Luís Orsi Capítulo redigido e publicado segundo as normas do periódico Check List: the journal of biodiversity data, disponível em: https://doi.org/10.15560/14.5.785 https://doi.org/10.15560/14.5.785 ANNOTATED LIST OF SPECIES Check List 14 (5): 785–799 https://doi.org/10.15560/14.5.785 Checklist of medium-sized to large mammals of Campos Gerais National Park, Paraná, Brazil Alan Deivid Pereira1, Sergio Bazilio2, Mário Luís Orsi3 1 Programa de Pós-Graduação em Ciências Biológicas, AC: Biodiversidade e Conservação de Habitats Fragmentados. Universidade Estadual de Londrina, Centro de Ciências Biológicas, Rodovia Celso Garcia Cid, PR 445, Km 380, CEP 86.057-970 – Londrina, Paraná, Brazil. 2 Universidade Estadual do Paraná – UNESPAR, Campus de União da Vitória. Caixa Postal 241, CEP 84600-970, União da Vitória, PR, Brazil. 3 Laboratório de Ecologia de Peixes e Invasões Biológicas. Universidade Estadual de Londrina, Centro de Ciências Biológicas, Departamento de Biologia Animal e Vegetal, Rodovia Celso Garcia Cid, PR 445, Km 380, CEP 86.057-970 – Londrina, Paraná, Brazil Corresponding author: Alan Deivid Pereira, alandeivid_bio@live.com Abstract Campos Gerais National Park lies within the Brazilian Atlantic Rainforest, a biodiversity hotspot and a priority for conservation. Current analysis, featuring a list of large and medium-sized mammal species in the park, was conducted between July 2013 and July 2014 and between May 2016 and April 2017. With a 780-hour sampling effort in active research and 157,516 hours in camera traps, 31 species of 17 families and 8 orders were recorded. Furthermore, 42% of recorded mammals in the park proved to be endangered species at state, national, or international levels. Two exotic and 1 domestic species were reported in the park. Results contribute towards an in-depth knowledge of the fauna in south Brazil and may help in further research work and management, complying with conservation proposals in the biodiversity of the Campos Gerais region in the state of Paraná, Brazil. Key words Atlantic Rainforest; conservation; inventories; Mixed Ombrophilous Forest; species richness; tropical forest. Academic editor: Átilla Colombo Ferreguetti | Received 19 May 2018 | Accepted 7 August 2018 | Published 28 September 2018 Citation: Pereira AD, Bazilio S, Orsi LM (2018) Checklist of medium-sized to large mammals of Campos Gerais National Park, Paraná, Brazil. Check List 14 (5): 785–799. https://doi.org/10.15560/14.5.785 Introduction The Brazilian Atlantic Rainforest Biome, which has only 12.4% of its original cover area, represented mostly by fragments less than 100 ha in area (SOS Mata Atlântica and INPA 2018), is a biodiversity hotspot and priority area for conservation (Myers et al. 2000). It is estimated that more than 298 mammal species are extant in the Atlantic Rainforest, with approximately 100 animals with an adult body mass of at least 1 kg, or rather, medium-sized and large animals (Paglia et al. 2012, Reis et al. 2014). Mammals have an important role in the maintenance and equilibrium of forest ecosystems (Miller et al. 2001, Magioli et al. 2015), with several ecological services. These comprise prey population control, plant pollination and seed dispersal, contributing towards the regeneration of forests (Terborgh et al. 1999, Galetti et al. 2015, Derhé et al. 2017). However, owing to anthropogenic forest fragmentation, modification of habitats, introduction of exotic species and other factors, several species of this group are endangered in many Brazilian states (MMA 2014, IUCN 2018). The above factors plus hunting Copyright Pereira et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. mailto:alandeivid_bio@live.com 786 Check List 14 (5) Figure 1. Map of the Parque Nacional dos Campos Gerais, state of Paraná, Brazil, with monitoring sites by camera traps during the 2013–2014 and 2015–2017 sampling periods. activities (Oliveira and Cassaro 2006) lead to loss of biodiversity, which cause the near extinction of several species (Mazzolli 2005). The establishment of conservation units comprises a type of action taken by the Brazilian government for the preservation of biodiversity (SNUC 2000). Conser- vation units have several aims, such as the protection of terrestrial and marine ecosystems, the safeguarding of endangered species at the regional and national levels and defending the region’s flora and fauna. In fact, conserva- tion should exist parallel to the aims of forest production (SNUC 2000). National Parks (PARNA in Portuguese) feature among the models of conservation units. In fact, the aim of PARNAs is to protect the nature, subsidiz- ing the natural sciences, allowing for tourist visits, and preserving forest and biodiversity for future generations (SNUC 2000). The state of Paraná, Brazil, is currently responsible for 5 National Parks, namely Parque Nacio- nal do Iguaçu, Parque Nacional do Superagui, Parque Nacional de Ilha Grande, Parque Nacional Saint-Hilaire/ Lange, and Parque Nacional dos Campos Gerais. The lat- ter was the last to be established (Oliveira 2012). Although the Parque Nacional dos Campos Gerais was established in 2006 to preserve remnants of the Mixed Ombrophilous Forest and native woods in the state of Parana (Oliveira 2012), no inventory of medium- sized and large mammals has been produced after 12 years. There is a knowledge gap for the Paraná Devonian Escarpment, as the region is scientifically called. Increasing changes in land use and occupation, and low representativeness of protected areas, rank the ecosystems of the Campos Gerais among the most endan- gered in the Brazil (Almeida and Moro 2007). Because crucial requirements for the development of conservation actions depend on basic knowledge of species and their distribution (Kasper et al. 2007), and the Campos Gerais study area lacks deep scientific information on biodiver- sity, especially of medium-sized and large mammals, the current analysis provides a list of medium-sized and large mammals for Parque Nacional dos Campos Gerais, Paraná, Brazil. Methods Study area. Parque Nacional dos Campos Gerais (here- after PNCG), in the southern state of Paraná, Brazil, has an area of 21,298.91 hectares, partially covering areas in the municipalities of Ponta Grossa, Castro, and Caram- beí. It lies on the Devonian Scarp region, with 2 sections on the first and second plateau of the Paraná (25°03.754 ́ S, 049°57.693ʹ W and 25°09.897ʹ S, 049°56.281ʹ W) (Oliveira 2012) (Fig. 1). Yearly rainfall ranges between 100 and 300 mm, with the mean temperature at 17.8 °C (Maack 2017). Soils are constituted of acric red-yellow latosol and dystrophic haplic cambisol (Almeida and Moro 2007). Remnants Pereira et al. | Checklist of mammals in Campos Gerais, Brazil 787 787 are frequently represented by almost circular patches of forests (regionally called “capões” in Portuguese), gal- lery forests or mixed woodland, particularly on slopes or diabase dikes (Moro 2001). The association between the Araucaria pine-tree forest and fields form the region’s typical landscape, combining significant forest areas and the last remnants of the Atlantic Rainforest (Maack 2017). Sampling. Sampling occurred at 2 different stages, namely, between July 2013 and July 2014, (permit 37691-1 ICMBio for scientific activities) and between May 2016 and April 2017 (permit 53800-1 ICMBio), totaling more than 780 hours of search in approximately 815 km of trails and roads. The following methods were applied: (1) direct search for evidences using the active search method (Voss and Emmons 1996), which consists of walking at an average speed of 1 km/h, on trails and dirt roads, searching for direct (e.g., sightings, vocaliza- tions) and indirect (e.g. footprints) evidences, and (2) camera traps. The sand and clay soil in the park did not require sand plots to record footprints. Temporal independence of the samples or counting the number of recorded footprints by a single researcher at intervals of at least 300 m between the first footprint up to the next one, was avoided. Continuous sequences on the same road were avoided, following Pardini et al. (2004). Surveys were monthly, starting at 8:00 h and lasting for 4–6 hours, depending on the number of records. Footprints were identified in the field and photographed. They were based on mea- surements and print format, confirmed later by specific literature on the theme (e.g. Becker and Dalponte 2013, Reis et al. 2014). Camera traps obtained information on nocturnal spe- cies and on those with difficult visualization, such as most medium-sized and large mammals (Srbek-Araujo and Chiarello 2013). Nineteen camera traps (Bushnell model) were distributed throughout the forest fragments, near water sprouts, on the treks and pathways of the park, taking into account the commonest trails used by spe- cies. Cameras were kept at a distance of at least 1 km and placed 50 cm above the ground on trees. The researchers also talked to local residents to complement information on their search on the species of the park. Camera traps were active during the 24 months of sampling, with a monthly change of memory cards and batteries during the study period. Sampling effort was equal to the number of camera traps multiplied by the number of sampling days (with 24 hours). An event was independent when there were (a) consecutive photographs by the same camera with an interval of at least 60 minutes and (b) non-con- secutive photographs by the same camera (Srbek-Araujo and Chiarello 2013). Observations of species were oppor- tunistically recorded in the study area to improve the species inventories obtained by camera-traps. Species were identified by camera trap records and opportunistic photos based on specialized literature (e.g. Oliveira and Cassaro 2006, Reis et al. 2011, 2014). Felines were identified by body size, pelage, nose, eyes and tail characteristics (for small felines) and by size and shape of footprint (for Puma concolor Linnaeus, 1771) (Oliveira and Cassaro 2006). All the species with an adult body mass equal of 1 kg or more were considered medium-sized and large mammals (Reis et al. 2011) and included in the list. Conservation status was obtained for each species within state, country and international context (BRASIL 2010, MMA 2014, IUCN 2018). Trophic guild classification followed Reis et al. (2014). The taxonomic nomenclature followed Paglia et al. (2012). Furthermore, the recent distinction between Leopardus guttulus (Hensel, 1872) and Leopardus tigrinus (Schreber, 1775) in southern and southeastern Brazil was acknowledged (Trigo et al. 2013). Patton et al. (2015) was consulted for xenarthrans and rodents. Data analysis. Records of footprints, camera traps and visual searches together made up the data for the list. Footprint records and photographs were separated to estimate expected richness according to different method- ologies. By separating footprint and camera-trap records, 2 rarefaction curves of mammal species were drawn, with 1,000 randomizations and first-order jackknife (Jackknife 1) estimator by EstimateS 8.2 (Colwell 2009). Results We recorded 31 medium-sized and large mammal species, distributed in 17 families and 8 orders (Figs 2–30; Table 1). Twenty-four species were recorded in 2013–2014 sampling, to which 7 additional species were added from the 2016-2017 survyes (Table 1). Twelve spe- cies belonged to the order Carnivora and 5 species from orders Rodentia and Cetartiodactyla, followed by the orders Cingulata, Didelphimorphia, Pilosa, Primates and Lagomorpha, which had 2 species each (Table 1). Species were distributed in 5 trophic guilds, or rather, 45.16% of the species were omnivorous, followed by carnivores, herbivores and frugivores (16.13% each) and insectivores (6.45%) (Table 1). Among the species recorded in current study, 11 (35.4%) were threatened at the state level; 8 (25.8%) at the national level; 6 species (19.3%) at the international level (Table 1). There were also 2 exotic species (Lepus europaeus (Pallas, 1778) and Sus scrofa Linnaeus, 1758) and a domestic one (Canis lupus familiaris) present in PNCG. Following the methodology in the current study, 54 footprint registrations were observed, with 10 species identified. Estimated richness was 12.7 ± 1.37 species (Fig. 31). Furthermore, 16 species were recorded by opportunistic observations and 2 species of primates were identified by vocalization (Table 1). Camera-trap sampling efforts from the 2013/2014 and 2016/2017 campaigns totaled 80,016 and 77,500 camera hours, respectively. Total sampling effort amounted to 788 Check List 14 (5) Figures 2–7. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 2. Didelphis albiventris. 3. Didelphis aurita. 4. Myrmecophaga tridactyla. 5. Tamandua tetradactyla. 6. Dasypus novemcinctus.7. Euphractus sexcinctus 157,516 camera trap hours and 26 recorded species with an estimated richness of 35 ± 1.56 (Fig. 31). Annotated list Didelphimorphia Didelphidae Didelphis albiventris Lund, 1840 Figure 2 Records. First record was in July 09, 2013, camera trap 6 (25°07.27ʹ S, 049°56.49ʹ W) and subsequently in the monitoring sites (camera traps 1, 2, 3, 4, 11, 12, 13, 14, 15, 16, 17, 19 and 19, see Fig. 1 for coordinates). Pereira et al. | Checklist of mammals in Campos Gerais, Brazil 789 789 Figures 8–13. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 8. Mazama gouazoubira. 9. Mazama americana. 10. Mazama nana. 11. Pecari tajacu. 12. Sus scrofa .13. Sapajus nigritus. Identification. Didelphis albiventris has totally white or black-tipped ears with whitish tips and no hair; face with yellowish-white coat with black spots around the eyes and a conspicuous stain on the central region of the head. Didelphis aurita (Wied-Neuwied, 1826) Figure 3 Records. First record was in August 06, 2013, camera trap 3 (25°03.59ʹ S, 049°57.26ʹ W) and subsequently in the monitoring sites (camera traps 1, 2, 9 and 11, see Fig. 1 for coordinates). Identification. Didelphis aurita has black hairless ears; variable head color, ranging from black to yellow. Black spots on the eyes and dark coloration on the dorsal region. 790 Check List 14 (5) Figures 14–19. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 14. Alouatta guariba clamitans.15. Cerdocyon thous. 16. Canis lupus familiaris. 17. Leopardus pardalis. 18. Leopardus guttulus. 19. Leopardus wiedii. Pilosa Mymercophagidae Myrmecophaga tridactyla Linnaeus, 1758 Figure 4 Records. First record was in August 08, 2016, camera trap 18 (25°03.37ʹ S, 049°59.44ʹ W) and subsequently in the same monitoring site. Identification. Two species of anteaters are known in the region of PNCG, with each having different body size and color pattern of pelage. Myrmecophaga tridactyla is a large species and weighs up to 45 kg. It has small ears and small eyes, long snout and an extremely long tongue; the thick pelage varies from dark gray and black, but with the paws white paws and some black bands at the top. The tail is robust and covered by hair. Pereira et al. | Checklist of mammals in Campos Gerais, Brazil 791 791 Figures 20–25. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 20. Puma concolor. 21. Puma yagouaroundi. 22. Eira barbara. 23. Galictis cuja. 24. Nasua nasua. 25. Procyon cancrivorus. Tamandua tetradactyla (Linnaeus, 1758) Figure 5 Records. First record was in August 08, 2016, camera trap 11 (25°03.04ʹ S, 049°59.33ʹ W) and subsequently in the same monitoring site. Identification. This is a smaller species, with an elon- gated snout and tongue (Fig. 5) and short, dense and pale-yellow pelage. There are 2 black stripes on the scap- ular region towards the posterior region of the animal. Cingulata Dasypodidae Dasypus novemcinctus Linnaeus, 1758 Figure 6 792 Check List 14 (5) Figures 26–30. Photographs of the medium-sized and large mammal species recorded in the Parque Nacional dos Campos Gerais, state of Paraná, Brazil. 26. Lepus europaeus. 27. Cuniculus paca. 28. Dasyprocta azarae. 29. Coendou spinosus. 30. Guer- linguetus brasiliensis. Records. First record was in July 10, 2013, camera trap 1 (25°02.25ʹ S, 050°01.39ʹ W) and subsequently in the monitoring sites (camera traps 2, 3, 4, 7 and 11, see Fig. 1 for coordinates). Identification. The species differentiation was based on characteristics of carapace that covers the body of the ani- mals. Species-based variations exist in the moving bands located in the median region of the body. Dasypus novem- cinctus has long, pointed ears, about 50% of the length of the head, which has a pinkish-yellow plaque (Fig. 6); dark carapace, with yellowish dermal shields. Although 9 mobile bands are extant, the number may range from 8 to 11. Euphractus sexcinctus Linnaeus, 1758 Figure 7 Pereira et al. | Checklist of mammals in Campos Gerais, Brazil 793 793 Table 1. List of land medium-sized and large mammal species at the Parque Nacional dos Campos Gerais, Brazil. Record method; Footprint (F), Visualization (VI), Camera traps (CT), Vocalization (VO). Conservation status by (IUCN), Brazilian List of Threatened Species (BR), Paraná state List of Threatened Species (PR). Data Deficient (DD), Endangered (EN), Least Concern (LC), Not evaluated (NE), Near Threatened (NT), Vulnerable (VU) and Critically Endangered (CR). Species added in 2016 to 2017 (+). Exotic species (*). Small species (**). Taxon Record Trophic guild Conservation status IUCN BR PR Didelphimorphia Didelphidae Didelphis albiventris Lund, 1840 VI-CT Omnivore LC NE LC Didelphis aurita (Wied-Neuwied, 1826) CT Omnivore LC NE LC Pilosa Mymercophagidae Myrmecophaga tridactyla Linnaeus, 1758+ CT Insectivore VU VU CR Tamandua tetradactyla (Linnaeus, 1758)+ CT Insectivore LC NE LC Cingulata Dasypodidae Dasypus novemcinctus Linnaeus, 1758 CT Omnivore LC NE LC Euphractus sexcinctus Linnaeus, 1758 CT Omnivore LC NE LC Cetartiodactyla Cervidae Mazama americana (Erxleben, 1777) VI-CT Herbivore DD NE VU Mazama gouazoubira (G. Fischer, 1814) F-VI-CT Herbivore VU VU VU Mazama nana (Hensel,1872)+ CT Herbivore LC NE LC Tayassuidae Pecari tajacu (Linnaeus, 1758) CT Omnivore LC LC VU Suidae Sus scrofa Linnaeus, 1758 *+ VI-CT Omnivore LC NE NE Primates Cebidae Sapajus nigritus Goldfuss, 1809 VI-VO Omnivore NT NE DD Atelidae Alouatta guariba clamitans (Humboldt, 1812) VI-VO Omnivore LC VU NT Carnivora Canidae Canis lupus familiaris Linnaeus 1758* F-VI-CT Omnivore NE NE NE Cerdocyon thous (Linnaeus, 1766) F-VI-CT Omnivore LC NE LC Chrysocyon brachyurus (Illiger, 1815)+ F-VI Omnivore NT VU VU Felidae Leopardus pardalis (Linnaeus, 1758) F-CT Carnivore LC NE VU Leopardus guttulus Hensel, 1872 CT Carnivore VU VU VU Leopardus wiedii (Schinz, 1821) CT Carnivore NT VU VU Puma concolor (Linnaeus, 1771) F-VI-CT Carnivore LC VU VU Puma yagouaroundi (É. Geoffroy Saint-Hilaire, 1803) VI-CT Carnivore LC VU DD Mustelidae Eira barbara (Linnaeus, 1758) CT Omnivore LC NE LC Galictis cuja (Molina, 1782)+ CT Omnivore LC NE NE Procyonidae Nasua nasua (Linnaeus, 1766) VI-CT Omnivore LC NE LC Procyon cancrivorus (G. [Baron] Cuvier,1798) F-CT Omnivore LC NE LC Lagomorpha Leporidae Lepus europaeus Pallas, 1778* F-VI Herbivore LC NE NE Rodentia Caviidae Hydrochoerus hydrochaeris (Linnaeus, 1766) F-VI Herbivore LC NE LC Cuniculidae Cuniculus paca (Linnaeus, 1766) F-CT Frugivore LC NE EN Dasyproctidae Dasyprocta azarae Lichtenstein, 1823 CT Frugivore DD NE LC Erethizontidae Coendou spinosus (F. Cuvier, 1823)+ VI-CT Frugivore LC NE LC Guerlinguetus brasiliensis (Gmelin,1788)** VI-CT Frugivore LC NE LC 794 Check List 14 (5) Figure 31. The rarefaction curve (observed and estimates by jackknife 1) of mammal species registered by identification of footprints and trap cameras. Vertical bars represent standard deviation. CT: Camera trap; F: footprints. Records. Record was in May 01, 2014 camera trap 1 (25°02.25ʹ S, 050°01.39ʹ W). Identification. Euphractus sexcinctus has 6 to 8 moving bands on the back of the carapace; there are 2 to 4 orifices in the region of the pelvic girdle near the base of the tail. Cetartiodactyla Cervidae Mazama americana (Erxleben, 1777) Figure 8 Records. First record was in July 26, 2013, camera trap 6 (25°07.51ʹ S, 049°57.08ʹ W) and subsequently in the monitoring sites (camera traps 5 and 11, see Fig. 1 for coordinates). Identification. The identification of species of the genus Cervidae was based on differences in body size, coat color and distribution area. Mazama americana is the largest species of the genus in Brazil, medium to mod- erately large, a factor that helps in the differentiation of the species. It has a reddish coloration, with white spots below the tail, inner face of limbs and ears. Mazama gouazoubira (G. Fischer, 1814) Figure 9 Records. First record was in July 30, 2013, camera trap 1 (25°02.25ʹ S, 050°01.39ʹ W) and subsequently in the monitoring sites (camera traps 2, 3, 5, 6, 7, 14 and 15, see Fig. 1 for coordinates). Identification. Mazama gouazoubira is a smaller species when compared to M. americana, weighing between 17 and 23 kg. The coloration is quite varied, with brown, gray and reddish variations. Mazama nana (Hensel, 1872) Figure 10 Records. Record was in May 17, 2016 camera trap 15 (25°03.36ʹ S, 049°59.44ʹ W). Identification. Mazama nana is the smallest species, with weight not exceeding 15 kg. The skin is character- ized by an intense bright reddish hue. Cetartiodactyla Tayassuidae Pecari tajacu (Linnaeus, 1758) Figure 11 Records. First record was in July 31, 2013, camera trap 6 (25°07.51ʹ S, 049°57.08ʹ W) and subsequently in the monitoring sites (camera traps 1, 2, 4, 7, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 and 19, see Fig. 1 for coordinates). Identification. The identification of the Brazilian native pig Pecari tajacu (Linnaeus, 1758) was based on pelage characteristics. A collar formed by slightly clear hair may be observed around its neck, differentiating the species from other native pigs. Suidae Sus scrofa Linnaeus, 1758 Figure 12 Records. First record was on May 5, 2013, camera trap 16 (25°03.37ʹ S, 049°58.43ʹ W) and subsequently in the monitoring sites (camera traps 1, 2, 4, 7, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 and 19, see Fig. 1 for coordinates). Identification. Sus scrofa is a swine native to Europe, Africa and Asia. Featuring great individual variations in body size and pelage color, it was introduced in the area under analysis. Although S. scrofa may be confused with P. tajacu, the species is usually larger. Primates Cebidae Sapajus nigritus (Goldfuss, 1809) Figure 13 Pereira et al. | Checklist of mammals in Campos Gerais, Brazil 795 795 Records. First record was in May 17, 2014, monitoring site 3 (25°03.59ʹ S, 049°57.26ʹ W) 12 individuals were recorded by opportunistic observations and subsequently in the monitoring sites 1, 2 and 11 (see Fig. 1 for coordinates). Identification. Sapajus nigritus has a generally black coloration, even though hue may vary by region. The face is pigmented, usually light brown, and the coat of the head is rather blacker. Atelidae Alouatta guariba clamitans Cabrera, 1940 Figure 14 Records. Record was in March 16, 2014, monitoring site 9 (25°12.12ʹ S, 049°56.39ʹ W) 7 individuals were recorded by opportunistic observations. Identification. Alouatta guariba clamitans is larger than S. nigritus. Males have a uniform reddish color, while the color of females ranges from blackish to dark-brown. The face is pigmented in darker shades of gray. Carnivora Canidae Cerdocyon thous (Linnaeus 1766) Figure 15 Records. First record was in October 12, 2013, camera trap 3 (25°03.59ʹ S, 049°57.26ʹ W) and subsequently in the monitoring sites (camera traps 1, 2, see Fig. 1 for coordinates). Identification. Cerdocyon thous was identified by its small size, and short, dense coat with a gray to brown hue. Chrysocyon brachyurus (Illiger, 1815) Records. Record was in May 12, 2016, monitoring site 11 (25°03.04ʹ S, 049°59.33ʹ W) 1 individual was recorded by opportunistic observations. Identification. Chrysocyon brachyurus is larger, with a small head in relation to body, small ears and tapering muzzle, with a general brown-orange coloration at the tip of the muzzle and end of the black limbs. The latter was only identified through footprints and personal reports of residents of farms near the PNCG. Canis lupus familiaris Linnaeus 1758 Figure 16 Records. First record was in July 13, 2013, camera trap trap 1 (25°02.25ʹ S, 050°01.39ʹ W) and subsequently in the monitoring sites (camera traps 3, 5, 7, 10, 12, 13, 14, 15, 16 and 19, see Fig. 1 for coordinates). Felidae Leopardus pardalis (Linnaeus, 1758) Figure 17 Records. First record was in October 14, 2013, camera trap 3 (25°03.59ʹ S, 049°57.26ʹ W) and subsequently in the monitoring sites (camera traps 5, 12, 13, 15 and 19, see Fig. 1 for coordinates). Identification. Five feline species were identified, based on patterns of body size and pelage. Leopardus pardalis was identified by its large body size, elongated rosettes pattern on the side of the body and a relatively smaller tail, when compared with other Leopardus species. Leopardus guttulus (Hensel, 1872) Figure 18 Records. First record was in January 28, 2014, camera trap 6 (25°07.51ʹ S, 049°57.08ʹ W) and subsequently in the monitoring sites (camera traps 5, 13, 14, 15, 17 and 18, see Fig. 1 for coordinates). Identification. Leopardus guttulus was identified by its relatively smaller tail with narrow rings, pink nose and pattern of circular shapes and eyespots on the sides of the body that differentiated the species from Leopardus wiedii. Leopardus wiedii (Schinz, 1821) Figure 19 Records. First record was in July 29, 2013, camera trap 1 (25°02.25ʹ S, 050°01.39ʹ W) and subsequently in the monitoring sites (camera traps 3, 4, 5, 9 and 12, see Fig. 1 for coordinates). Identification. Leopardus wiedii is characterized by a long tail, large eyes, protruding muzzle and large paws. Puma concolor (Linnaeus, 1771) Figure 20 Records. First record was in December 09, 2013, camera trap 2 (25°08.02ʹ S, 049°56.17ʹ W) and subsequently in the monitoring sites (camera traps 3, 11, 13, 14, 15, and 16, see Fig. 1 for coordinates). Identification. Puma concolor is distinguishable by its short and uniform coat of brown color, ranging from light to reddish tones. Puma yagouaroundi (É. Geoffroy Saint-Hilaire, 1803) Figure 21 Records. First record was in October 11, 2013, camera trap 5 (25°07.27ʹ S, 049°56.49ʹ W) and subsequently in the monitoring sites (camera trap 7, see Fig. 1 for coor- dinates). Identification. Puma yagouaroundi was identified by its monochromatic long body and tail. Mustelidae Eira barbara (Linnaeus, 1758) 796 Check List 14 (5) Figure 22 Records. First record was in July 17, 2013, camera trap 3 (25°03.59ʹ S, 049°57.26ʹ W) and subsequently in the monitoring sites (camera traps 1, 4, 5, 12, 13, 15, 16, 18 and 19, see Fig. 1 for coordinates). Identification. Two mustelids were registered in the PNCG. The identification of these animals was based on the pattern of the 2 species, namely, long body, short limbs and long tail. In the case of Eira barbara the color of the coat is dark brown throughout the entire body, with a lighter brown for head and neck, with regional variations. Galictis cuja (Molina, 1782) Figure 23 Records. First record was in May 03, 2017, camera trap 18 (25°03.37ʹ S, 049°59.44ʹ W) and subsequently in the monitoring site 11 (see Fig. 1 for coordinates). Identification. Galictis cuja is smaller in size than E. barbara; paws, belly, throat and face are black, with a yellowish back. Procyonidae Nasua nasua (Linnaeus, 1766) Figure 24 Records. First record was in July 13, 2013, camera trap 1 (25°02.25ʹ S, 050°01.39ʹ W) and subsequently in the monitoring sites (camera traps 2, 3, 4, 5, 6, 12, 13, and 15, see Fig. 1 for coordinates). Identification. Two species of the family Procyonidae were registered. Nasua nasua, an exclusive species of South America, has an enlarged head tapering into a narrow and prolonged pointed snout. Coloration varies according to group, although the general pattern ranges between light and dark brown, with a generally striped tail. Procyon cancrivorus (G. Cuvier, 1798) Figure 25 Records. First record was in August 23, 2013, camera trap 3 (25°03.59ʹ S, 049°57.26ʹ W) and subsequently in the monitoring sites (camera traps 6 and 18, see Fig. 1 for coordinates). Identification. Procyon cancrivorus is easily identified by its black mask that descends from the eyes to the base of the mandible; its tail is characterized by several dark rings. Lagomorpha Leporidae Lepus europaeus (Pallas, 1778) Figure 26 Records. First record was in October 07, 2013, monitor- ing site 3 (25°03.59ʹ S, 049°57.26ʹ W) individuals were recorded by opportunistic observations and subsequently in the monitoring sites 7, 8 and 18 (see Fig. 1 for coor- dinates). Identification. Lepus europaeus is a species of European origin introduced in South America. It is larger in size than the native Sylvilagus brasiliensis (Linnaeus, 1758), with long ears and legs; general brownish gray color on the upper parts and of a somewhat lighter hue on the lower part. The above characteristics distinguish the hair of S. brasiliensis. Rodentia Caviidae Hydrochoerus hydrochaeris (Linnaeus, 1766) Records. Record was in November 29, 2013, monitoring site 7 (25°09.42ʹ S, 049°59.44ʹ W). Identification. Hydrochoerus hydrochaeris is the largest rodent in the world, easily identifiable due to the shape of the head with short ears and hue ranging between reddish to grayish brown. The species was recorded based on the analy- sis of footprints and opportunistic observations (Table 1) Cuniculidae Cuniculus paca (Linnaeus, 1766) Figure 27 Records. First record was in July 12, 2013, camera trap 5 (25°07.27ʹ S, 049°56.49ʹ W) and subsequently in the monitoring sites (camera traps 1, 2, 3, 6, 7, 8, 9, 11, 12, 13, 17, 18 and 19, see Fig. 1 for coordinates). Identification. Cuniculus paca is a medium-sized rodent. Identification of the species was based on the pattern of the coat, ranging between brownish-red and dark brown, with a pattern of whitish rounded spots in longitudinal lines. Dasyproctidae Dasyprocta azarae Lichtenstein, 1823 Figure 28 Records. First record was in July 12, 2013, camera trap 5 (25°07.27ʹ S, 049°56.49ʹ W) and subsequently in the all monitoring sites (see Fig. 1 for coordinates). Identification. Dasyprocta azarae is a large orange- brown rodent with a rounded back and long skinny legs. Although 9 species of the genus Dasyprocta have been described for Brazil, D. azarae is the only species with a record of occurrences for the state of Paraná. Erethizontidae Coendou spinosus (F. Cuvier, 1823) Figure 29 Pereira et al. | Checklist of mammals in Campos Gerais, Brazil 797 797 Records. Record was in May 22, 2016 camera trap 12 (25°02.55ʹ S, 049°57.41ʹ W) and by opportunistic obser- vations in the same monitoring site. Identification. Coendou spinosus was identified by coat and body shape. The coat is formed by a mixture of rigid, aculeiform (cylindrical ‘spines’) hairs and finer hairs. The former are longer than the latter ones. Coloration varies from yellowish to dark brown at the back. Sciuridae Guerlinguetus brasiliensis (Gmelin, 1788) Figure 30 Records. First record was in February 02, 2014, camera trap 5 (25°07.27ʹ S, 049°56.49ʹ W) and subsequently 2 individuals were recorded by opportunistic observations in the monitoring sites 1 and 7 (see Fig. 1 for coordinates). Identification. The squirrel Guerlinguetus brasiliensis may be included among the native species. Although small in size, it is easily identifiable through photographs. The species is an average sized squirrel with occurrences recorded in the Atlantic Rainforest. It is distinguished from other squirrels due to its intermediate body size and such characteristics as voluminous tail, longer than or equal to the body. Discussion Because Parque Nacional dos Campos Gerais has the largest phytophysionomy area of the Campos Gerais in Paraná (Maack 2017), it is highly important for the conservation of biodiversity. In fact, it harbors endemic and threatened species within its borders (Oliveira 2012). Species richness recorded by us corresponds to 56% of all medium-sized and large mammals with current occurrence for the state of Paraná (Reis et al. 2009) and approximately 31% of medium-sized and large mammals for the Atlantic Rainforest (Paglia et al. 2012, Reis et al. 2014). Although there is a stabilization of the spe- cies accumulation curve in our study (Fig. 34), other species may also occur in the park. In fact, sampling in 2016– 2017 also included M. tridactyla, T. tetradactyla, M. nana, S. scrofa, C. brachyurus, G.brasiliensis, and C. spinosus to the final list. According to Srbek-Araujo and Chiarello (2007), periods longer than 250 days are suf- ficient for a faunal survey. The species richness recorded in the PNCG is similar to that observed in other studies carried out in conser- vation units nearby. Borges (1989) reported 40 species of medium- sized to large mammals in Parque Estadual de Vila Velha, which is located 20 km from PNCG. In Reserva Biológica das Araucarias some 56 km from PNCG, the richness amounted to 28 species (D’Bastiani et al. 2018). In the Floresta Naciona de Irati, some 70 km from PNCG, 24 species of medium-sized to large mam- mals were recorded (Pereira and Bazilio 2014). Species such as M. nana, C. brachyurus, and M. tridactyla were not recorded at the Reserva Biológica das Araucarias and Floresta Naciona de Irati. Although these species have been reported by Borges (1989) for Parque Estadual de Vila Velha, he also included in his records sampling areas that are now part of PNGC. The order Carnivora, with 12 species, is the order most recorded in ourstudy. According to Chiarello (2000), the group occurs with frequency in fragmented forest remnants. Species of Carnivora have great mobility and capacity in exploring man-disturbed environments close to native vegetation (Lyra-Jorge et al. 2010). Chrys- ocyon brachyurus was recorded by footprints and from accounts by farm workers near the park, is the largest South American canid, with adults weighing between 20 and 30 kg (Rodden et al. 2004). Although C. brachyurus is found mainly in the Campos Gerais region, it is there has been a reduction in its population numbers over the last decades. In fact, it has become scarce (Cherem and Perez 1996, Bazilio et al. 2011). Five of the 7 species of felines in Paraná (Reis et al. 2009) occurred in the study area, specifically L. guttulus, L. pardalis, L. wiedii, P. yagouaroundi, and P. concolor. Our results corroborate those by Bastiani et al. (2015), who recorded practically the same species in the Floresta Nacional de Piraí (a 150 ha fragment some 83km dis- tance from PNCG). The low detection of several species of felines, such as L. guttulus and L. wiedii, may be due to competition for resources with large felines (Oliveira et al. 2010). The presence of medium-sized and large mammals carnivore and herbivore species in the area studied indicates that the characteristics of the forest fragment enhance ecological processes for its maintenance. Her- bivore animals disperse and control plant populations, whilst carnivores maintain herbivore populations (Santos et al. 2004, Pardini et al. 2004). Of the threatened species in the PNCG, the presence of M. americana, M. nana, P. tajacu and C. paca is espe- cially important because of their high conservation status in the state of Parana, which is linked to illegal hunting and trade in pelts (Cullen et al. 2000). PNCG is offers protection for these and other species. According to Mazzolli (2006), P. tajacu is indicative of environmental quality because it has a low tolerance for disturbed habitats, and as a corollary, its absence is suggestive of highly disturbed habitat. Desbiez et al. (2012) demonstrated that the overlap in food resources and habitat use in Brazil between feral pigs and Tayassu pecari (Link, 1795) were lower than expected. In the Brazilian Pantanal, Galetti et al. (2015) found a great overlap in diets of T. pecari and feral pigs, but less over- lap between P. tajacu and feral pigs. Nevertheless, feral pigs may impact the natural community in several other ways, such as eating eggs, destroying by rooting, and serving as vectors for disease (Desbiez et al. 2012, Galetti et al. 2015). 798 Check List 14 (5) The wild boar is in the list of 100 invading and heav- ily impacting species worldwide (Lowe et al. 2000). Loss of biodiversity and the extinction of native species caused by the introduction of wild pigs have already been documented (Wolf and Conover 2003). Competitive interaction with the wild boar mainly excludes peccaries (Gabor and Hellgren 2000, Galetti et al. 2015). At least 4 java pigs have been reported in the Alagadas Reservoir in northwestern PNCG. Piglets with spots characteristic of the java pig are common among other piglets with spots or common features of domestic pigs. The domestic dog (C. lupus familiaris) has been recorded on trails and roads, as well as within forests in PNCG. The presence of domestic animals may have serious ecological consequences for native fauna in conservation units (Rangel and Neiva 2013, Doherty et al. 2017). The predation of wild fauna by almost savage domestic dogs is compounded by direct competition for resources with native carnivores (Galetti and Sazima 2006). Exotic species are nowadays acknowledged to be the secondmost important threat to biodiversity, but also cause economic losses and pose serious risks to human health (Dorcas et al. 2012). In the wake of processes that alter habitat, knowledge of biodiversity in conservation units is a basic require- ment for management plans, proposals for conservation, and studies on ecological patterns and species distribu- tion, (Silveira et al. 2010, Oliveira et al. 2017). Our study demonstrates the importance of PNCG as a haven for Paraná’s medium-sized and large mammals. However, the occurrence of exotic and domestic species and the presence of hunters in the conservation unit underscore the need for more surveillance and monitoring, the repression of hunting, the control of exotic species, and the development of environmental education within the local community . Acknowledgements We thank the ICMBio team of the Parque Nacional dos Campos Gerais for its support in our research. Thanks are due to Juliane Coimbra Bczuska, Denise Bener, and Elvira D’Bastiani for their assistance in data collection. Thanks are also due to the Coordination for the Upgrad- ing of Higher Education Personnel (Capes), the Araucária Foundation, and the Fundation Boticário for the Protec- tion of Nature (in our second sampling year) for funding. References Almeida CG, Moro RS (2007) Análise da cobertura florestal no Parque Nacional dos Campos Gerais, Paraná, como subsídio ao seu plano de manejo. Terr@ Plural 1 (1): 115–122. Bastiani E, Bazilio S, Barros KF, Nabrzecki G (2015) Felinos da Floresta Nacional de Piraí do Sul, Paraná–Brasil. Acta zoológica mexicana 31 (1): 23–26. Bazilio S, Schemczssen Z, Marques AC (2011) Registro visual do lobo- -guará, Chrysocyon brachyurus (Illiger, 1815) (Mammalia: Carni- vora: Canidae) na Floresta Nacional de Três Barras, SC. 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Capítulo redigido segundo as normas do periódico Papéis Avulsos de Zoologia data, disponível em: http://www.revistas.usp.br/paz/about/submissions#authorGuidelines http://www.revistas.usp.br/paz/about/submissions#authorGuidelines 41 Narrow, short and deadly: Mammals roadkills on highway sections of PR-445, South of Brazil Running Title: Mammals roadkills on highway sections of PR-445 Alan Deivid Pereira¹* https://orcid.org/0000-0002-3182-2344 Marcelo Hideki Shigaki Yabu² https://orcid.org/0000-0002-9931-9008 Iago Vinicios Geller¹ https://orcid.org/0000-0003-2838-8724 Carolina Prado² https://orcid.org/0000-0002-6477-6962 Carlos Rodrigo Lehn¹ https://orcid.org/0000-0003-2865-1019 Ana Paula Vidotto-Magnoni² https://orcid.org/0000-0003-1819-7019 Juliano André Bogoni3https://orcid.org/0000-0002-8541-0556 Mário Luís Orsi²https://orcid.org/0000-0001-9545-4985 1 Universidade Estadual de Londrina, Programa de Pós-Graduação em Ciências Biológicas, Rodovia Celso Garcia Cid, PR 445, Km 380, Campus Universitário, CP 10.011, CEP 86057-970, Londrina, PR, Brazil. 2 Universidade Estadual de Londrina, Laboratório de Ecologia de Peixes e Invasões Biológicas, Rodovia Celso Garcia Cid, PR 445, Km 380, Campus Universitário, CP 10.011, CEP 86057-970, Londrina, PR, Brazil. 3 Universidade de São Paulo, Escola Superior de Agricultura “Luiz de Queiroz”, Laboratório de Ecologia, Manejo e Conservação de Fauna Silvestre (LEMaC), Piracicaba, São Paulo, Brazil. https://orcid.org/0000-0002-3182-2344 https://orcid.org/0000-0002-9931-9008 https://orcid.org/0000-0003-2838-8724 https://orcid.org/0000-0002-6477-6962 https://orcid.org/0000-0003-2865-1019 https://orcid.org/0000-0003-1819-7019 https://orcid.org/0000-0002-8541-0556 https://orcid.org/0000-0001-9545-4985 42 ABSTRACT Animal-vehicle collisions are the main negative impact of roads on wildlife, where they cause population declines, changes in the structuring of communities, and potential changes in species behavior. Here, we determined mammal roadkill rates and the hotspots with higher rates for medium- and large-bodied mammals on Highway PR-445 in the state of Paraná, Brazil. We also evaluated possible differences in the frequency of roadkills concerning the activity pattern and feeding habit of species. In doing so, we monitored PR-455 weekly on average twice a week from March 2018 to March 2019, totaling 7296 km traveled during 96 trips over 12 months. We recorded 60 roadkill mammals belonging to 17 species, representing a rate of 0.151 individual/km/day. The orders Carnivora, Cingulata, and Didelphimorphia showed the most common roadkills. Omnivores were more prone to getting hit by vehicles than herbivorous and carnivorous species. The type of mammal activity pattern was not a determinant in explaining the differences in roadkill rates. Highways in Paraná are among the roads that register the most vertebrate-vehicle collisions in the country. This issue, together with extensive habitat conversion and fragmentation, increases the threats to the relictual fauna. Our results indicate that the regions with the highest incidence of roadkills on PR-445 are those close to stretches over rivers and with remnants of native vegetation. Thus, we emphasize that more comprehensive measures (e.g., wildlife passages and speed reducers) are essential to mitigate the impact of roads on wildlife. Keywords: Animal-vehicle collisions; mammals; activity period; species diet; diversity loss 43 INTRODUCTION In the 21st century, roads account for some of the most important environmental changes resulting in a high impact on natural landscapes (Laurance et al., 2014). These impacts include an increase in noise and light pollution, erosion, and loss of natural environment and the character of the land, aggravating the edge effect of forest fragments (Forman & Alexander, 1998; Orlandin et al., 2015). The consequences of roads on wildlife reach different ecological scales, including changes in the structure of communities (Laurance et al., 2009) and behavioral dynamics (Ascensão et al., 2014; Poessel et al., 2014), influencing the presence and persistence of species in surrounding forest fragments (Borda-de- Água et al., 2011) and also causing considerable economic losses (Abra et al., 2019). Besides the abovementioned road-related threats to animals, roadkill is the main negative impact of roads on wildlife, with direct effects on local populations, influencing species abundance and distribution (Eigenbrod et al., 2008). Population declines caused by road impacts have effects on the variability and genetic viability of species (Jackson & Fahrig, 2011). Roadkill is the primary cause of death for some species, including threatened species (Forman et al., 2003), proving to be a direct factor of mortality among terrestrial vertebrates as significant as hunting (Seiler & Heldin, 2006). Brazil has one of the most extensive road networks in the world, covering more than 200,000 km of surfaced roads (ANTT, 2019). According to the estimates provided by the Brazilian Center for Road Ecology Research (CBEE), more than 15 animals die on Brazilian roads every second. It is estimated that 5 million medium- and large-bodied animals such as giant anteaters, peccaries, deer, jaguars, monkeys, and maned wolves are killed annually on the roads and highways of Brazil (CBEE, 2019). Therefore, road monitoring and implementation of mitigation strategies are essential to minimize the harm of road to native biota (Costa et al., 2015; Abra et al., 2019). This action is even more crucial in areas of great value for biodiversity conservation, such as the Atlantic Forest biome. In the northern part of the state of Paraná, the predominant vegetation is seasonal semideciduous forest (SSF), which is one of the most rarefied ecosystems in the Atlantic Forest occurring in refuge areas of South and Southeast Brazil (SOS Mata Atlântica & INPE, 2018). Recent estimates have shown 44 that only 7% of the area covered initially by SSF remains, being represented by small fragments (typically less than 50 ha), immersed in matrices of areas of intensive farming and urban areas crisscrossed by extensive road networks with intense traffic (SOS Mata Atlântica & INPE, 2018). Within Paraná, studies involving mammal roadkills have been conducted in the surroundings of Iguaçu National Park (Brocardo & Cândido-Junior, 2012), in the coastal region (i.e., Serra do Mar) (Leite et al., 2012; Belão et al., 2014) and in the region of Campos Gerais (i.e., central plateaus) (Zeliski et al., 2009; Weiss & Vianna, 2012). However, no studies have extensively explored the impact of roads on mammalian fauna in the northern region. This meta-region represents an important interest in biodiversity conservation owing to its location in a transition zone between the tropical and subtropical portion of the Atlantic Forest. Evaluating the importance of animal-vehicle collisions is a complex challenge in conservation ecology, involving ecological, economic, social and technical perspectives, and considering both wide-ranging and small spatial scales (Seiler & Heldin, 2006). The information collected from road data surveys is often used to choose the best strategies for reducing collisions with animals and to evaluate their effectiveness (Cureton & Deaton, 2012; Lesbarreres & Fahrig, 2012; Costa et al., 2015). Thus, this study determined roadkill rates and pointed out the hotspots with a higher incidence of roadkills of medium- and large-bodied mammals along Highway PR-445, located in the subtropical portion of the Atlantic Forest in South Brazil. We also evaluated the possible variation in the frequency of roadkills according to the activity pattern and feeding habit of mammalian species. MATERIAL AND METHODS Study area Highway PR-445 (also called Celso Garcia Cid) is 95 km long, located between the municipalities of Mauá da Serra and Warta in northern Paraná (23.30°S, 51.17°W). This highway is marked by being located in the transition zone between the tropical and subtropical Atlantic Rainforest (Tropic of Capricorn, 23.26ºS). It is a state-administered 2-lane highway with a short 4-lane section in the city of 45 Londrina. The maximum speed is 80 km/h, being controlled by electronic radar and mobile surveillance. The metropolitan region of Londrina is the fourth largest metropolis in southern Brazil, with an estimated population of more than one million, and it has heavy traffic. Traffic with heavy vehicles (e.g., ≥ 3500 kg) is constant mainly due to the transport of crops during harvest time. The predominant vegetation in the region is seasonal semideciduous forest. The climate of the region is classified as humid subtropical mesothermal (Cfa), with an annual average temperature around 21°C, and an annual rainfall of 1450 mm (Peel et al., 2007). Highway PR-445 is surrounded mainly by an agricultural matrix (soybean, wheat and corn crops), eucalyptus monocultures, pastures and urban areas, along with small native remnants of SSF (SOS Mata Atlântica & INPE, 2018). Sampling The data on mammal roadkills were obtained between kilometers 1 and 76 of PR-445 (80% of the whole road), which includes the counties of Londrina, Mauá da Serra and Tamarana (Fig. 1). We adopted a monitoring protocol according to Costa et al. (2015), using mixed protocols (monthly and weekly samplings) to achieve an inventory of the species with no marked peak mortality (i.e., species with higher activity in warmer periods [southern hemisphere summer]). 46 Figure 1. Location of Highway PR-445 in the state of Paraná. Brazil indicating the sector measured in this study from 2018 to 2019. The hotspots highlighted in red indicate places with high rates of mammal roadkill. We monitored PR-445 on average twice a week, from March 2018 to March 2019, totaling 7296 km on 96 trips over 12 months. The search for medium- to large-bodied mammal carcasses was done by car in the morning period (from 7:00 a.m. to 12:00 p.m.) with an average speed of 80 km/h. The roadkill mammals found were photographed and identified in situ whenever possible. Accordingly, we used specific field guides for the taxonomic groups (Reis et al., 2009, 2014). For the carcasses with advanced decomposition or showing damage that hindered accurate species identification, we collected samples of hairs for species identification on slides based on microstructural characteristics, namely cuticular and medullary patterns. The hairs were collected manually from each carcass. We extracted small tufts of guard hairs from the back at the intersection of the median line and scapular waistline, following the identification protocol described by Quadros & Monteiro-Filho (2006). Species identification was confirmed using the identification keys of Quadros & Monteiro-Filho (2010) and Miranda et al. (2014). 47 The alpha taxonomic nomenclature followed Paglia et al. (2012) and considering recent taxonomic changes (e.g., Leopardus guttulus (Hensel, 1872) based on Trigo et al. (2013) and in the genus Sapajus according to Lych-Alfaro et al. (2012). For the orders Pilosa and Rodentia we also consulted Patton et al. (2015) for accurate identification. All species with an adult body mass ≥1 kg was considered medium-to large-bodied mammals (Paglia et al., 2012). Mammalian activity pattern was defined as follows: (1) crepuscular/nocturnal (CrN), (2) diurnal (D) and (3) nocturnal (N) (Reis et al., 2011). Feeding habits were according to Paglia et al. (2012), as follows: (i) carnivore (Ca), (ii) frugivore and folivore (Fr/Fo), (iii) frugivore and omnivore (Fr/On), (iv) herbivore or grazer (Hb), (v) insectivore and omnivore (In/On) and (vi) myrmecophage (Myr). Moreover, for each species, we obtained the conservation status according to the Brazilian Wildlife Red Book (MMA, 2018), the List of Threatened Fauna in the State of Paraná (Paraná, 2010) and IUCN Red List of Threatened Species (IUCN, 2019). Data analysis The location of each animal was recorded using a Global Positioning System (GPS). To analyze the regions of the highway with the highest concentration of roadkills (i.e., hotspots), we created a density map of records. Accordingly, we used the kernel density estimator. The kernel approach allows estimating the number of events per unit area in each cell of a regular grid covering the study area, in addition to filtering the variability of a dataset (Wand & Jones, 1995). The analysis was performed using software R based on the "MASS version 2.23-15" package and "KernSmooth" function (Wand, 2015). The frequency (F) of roadkills for each recorded species was calculated by the formula: 𝐹𝑖 = ( 𝑁𝑖 𝑁 ) 100 where: Fi = frequency of occurrence for the ith species; Ni = number of records of the ith species; N = total number of records. 48 We used the non-parametric test of Kruskal-Wallis, the non-parametric equivalent of ANOVA, due to non-normal data determined a priori by frequency histogram and Shapiro-Wilk test (Shapiro & Wilk 1965) to evaluate possible differences in the number of records in about activity patterns and feeding habits. Since we identified a statistically significant difference, we performed pairwise comparisons using the Tukey and Kramer (Nemenyi) test with Tukey-Dist approximation for independent samples (Nordstokke & Stelnicki, 2014). All analyses were performed using software R based on the "Vegan version 2.4-1" package (Oksanen et al., 2019). RESULTS We recorded 60 specimens of mammal roadkill on PR-445, resulting in 17 species distributed in 12 families and 7 orders (Table 1), corresponding to a roadkill rate of 0.151 individual/km/day-. The kernel density estimator indicated three points along the highway with the highest concentration of records of roadkill animals, i.e., hotspots. One hotspot was between Mauá da Serra and Tamarana (km 4; 23º 83 S, 51º 16 W), a stretch near the bridge over the Taquaruna River (Km 40; 23º 56 S, 51º11 W), and another eight kilometers perpendicular to the conservation unit Parque Estadual Mata dos Godoy, towards the urban area of Londrina (km 58; 23º 43 S, 51º13 W) (Fig. 1). The higher numbers of roadkill events were found for the orders Carnivora (48.3% of roadkill records), Cingulata (16.7%) and Didelphimorphia (15%). The other roadkill records included the orders Lagomorpha (6.7%), Primates and Rodentia (5% each) and Myrmecophagidae (3.3%). Of the 17 species recorded, five are included in a category with some degree of threat, from the state to international level according to the criteria established by the IUCN (Table 1). Table 1. List of roadkill medium- to large-bodied mammalian species on Highway PR-445. Acronyms are as follows. N: Total number of occurrences. F: Frequency of occurrence by species. Diet: carnivore (Ca); frugivore and folivore (Fr/Fo); frugivore and omnivore (Fr/On); herbivore or grazer (Hb); insectivore and omnivore (In/On) and myrmecophage (Myr). Activity pattern: crepuscular/nocturnal (CrN); diurnal (Dn); nocturnal (Nt). Threat category: data deficient (DD); endangered (EN); least 49 concern (LC); not evaluated (NE); near threatened (NT) and vulnerable (VU). Exotic species (*). Small- bodied species (**). 50 Taxon Number of records Diet Activity pattern Threat category N F IUCN BR PR Didelphimorphia Didelphidae Didelphis albiventris Lund, 1840 7 11.7 Fr/On CrN LC NE LC Didelphis aurita Wied-Neuwied, 1826 2 3.3 Fr/On Nt LC NE LC Pilosa Myrmecophagidae Tamandua tetradactyla (Linnaeus, 1758) 2 3.3 Myr Nt LC NE NE Cingulata Dasypodidae Dasypus novemcinctus Linnaeus, 1758 10 16.7 In/On CrN LC NE LC Primates Atelidae Sapajus nigritus (Goldfuss, 1809) 3 5 Fr/On Dn NT NE DD Carnivora Canidae Cerdocyon thous (Linnaeus, 1766) 13 21.7 In/On CrN LC NE LC Felidae Leopardus guttulus (Hensel, 1872) 3 5 Ca Nt VU VU VU Leopardus wiedii (Schinz, 1821) 1 1.7 Ca Nt NT VU VU Puma yagouaroundi (E. Geoffroy, 1803) 1 1.7 Ca Dn LC VU DD Mustelidae Eira barbara (Linnaeus, 1758) 1 1.7 Fr/On Dn LC NE LC Galictis cuja (Molina, 1782) 1 1.7 Ca CrN LC NE LC Procyonidae Nasua nasua (Linnaeus, 1766) 9 15 Fr/On Dn LC NE LC Lagomorpha Leporidae Lepus europaeus Pallas, 1778* 2 3.3 Hb CrN LC NE NE Sylvilagus brasiliensis (Linnaeus, 1758) 2 3.3 Hb CrN LC NE VU Rodentia Cuniculidae Cavia aperea Erxleben, 1777** 1 1.7 Hb Nt LC NE LC Erethizontidae Coendou spinosus (F. Cuvier, 1823) 1 1.7 Fr/Fo Nt DD NE LC Myocastoridae Myocastor coypus (Molina, 1782)* 1 1.7 Fr/On Nt LC NE LC 51 Based on the analysis of the cuticular and medullary patterns of mammal hair microstructure, we were able to perform an accurate identification of feline species (Fig. 2). We found three cuticular patterns, namely wide rhombus (Puma yagouaroundi), narrow rhombus (Leopardus guttulus) and narrow leaf shape (Leopardus wiedii), and also two medullary patterns, namely trabecular fimbriate with vacuoles in P. yagouaroundi and L. guttulus, and trabecular fimbriate without vacuoles for L. wiedii (Fig. 2). Figure 2. Microstructural pattern of species of the family Felidae: Leopardus wiedii. (A) cuticular pattern and (B) medullar pattern; Leopardus guttulus; (C) cuticular pattern and (D) medullar pattern; and Puma yagouaroundi (E) cuticular pattern and (F) medullar pattern. We did not find significant differences in the frequency of roadkills with regard to activity pattern of mammals [H = 3.2919; DF = 2; p > 0.05] (Fig. 3A). However, we found significant differences in the frequency of roadkills with regard to feeding habit [H = 6.869; DF = 5; p < 0.05] (Fig. 3B). 52 Figure 3. Boxplot illustrating the difference in roadkill rate for medium- to large-bodied mammals on Highway PR-445 between the municipalities of Londrina and Mauá da Serra, PR in relation to: (A) activity pattern; and (B) feeding habit. Acronyms are described in Table 1. A: a significant difference [p= 0.05] is shown between carnivores (CA) and frugivores/omnivores (Fr/On). B: a significant difference [p = 0.04] is shown between carnivores (CA) and insectivores/omnivores (In/On). DISCUSSION Direct mortality is the most widespread and significant effect of roads on wildlife (Bissonette, 2002), which can be evidenced throughout our study. However, the number of roadkills presented here may be underestimated, since larger animals may not die immediately after the collision, moving to areas away from the road, making it difficult to include them in the count (Bruinderink & Hazebroek, 1996). 53 We found a rate of 0.151 dead animals per day for each kilometer traveled along PR-445. Considering this crude rate, >5200 mammals succumb during a year on this highway alone, and at this same rate applied to the entire network of paved roads in Brazil, the number of roadkill mammals is as high as 11.7 M individuals. However, the rate determined in our study is higher than the value of 0.017 found for the PR-406 and PR-508 highways in the coastal region of Paraná (Leite et al., 2012). Even studies of short stretches of highway have been shown to have a similar significant impact on wildlife as those of longer stretches (Magioli et al., 2019). The high roadkill rate for many species is appalling and indicates that these accidents are commonplace and widespread (Seiler & Heldin, 2006). Yet, the Brazilian Atlantic Forest has high rates of defaunation of medium- to large-bodied mammals (Bogoni et al., 2018), especially in highly fragmented areas of the present study, so that the impact of roads on wildlife is a crucial factor to be considered in conservation strategies. A series of empirical studies showed that roads have substantial impacts on species abundance and species composition in ecosystems (Polak et al., 2014). Among the roadkilled species recorded in our study, 43% belonged to the order Carnivora. Presumably, the high number of Carnivora roadkills may be related to their ecological and behavioral aspects. Species of the order Carnivora depend on a large area to prosper, since having large home ranges (a powerful descriptor of spatial requirements of species or populations). Moreover, Carnivora species show the ability to explore human-disturbed environments neighboring native vegetation (Lyra-Jorge et al., 2010). Studies indicate that roads affect the movement and wide distribution of carnivores, including felids, particularly those living in habitats near urbanized areas (Poessel et al., 2014). The population declines of carnivores due to roadkills compromises the stability of forests and ecosystem functioning. Failure to thrive in landscapes highly modified by humans leads to non-random changes that propagate throughout all communities and ecosystems (i.e., cascade effects) because Carnivora animals play an important role in providing ecosystem services (e.g., disease control), being considered apex predators or mesopredators in food chains, contributing to the regulation of other populations (Terborgh et al., 1999). 54 The most frequent roadkilled mammals were particularly Cerdocyon thous, Dasypus novemcinctus, Nasua nasua and Didelphis albiventris. In the Atlantic Forest, these species are easily found in altered environments and a have preference for open areas (Reis et al., 2014). Although C. thous does not appear in the Brazil Red Book of Threatened Species of Fauna (MMA, 2018), the high roadkill rates for this species is cause for concern regarding its conservation, since in many studies C. thous leads statistics for species most affected by roadkills throughout Brazil (Zeliski et al., 2009; Cunha et al., 2010; Belão et al., 2014; Brum et al., 2018; Zanzini et al., 2018). The other species did not have high frequencies of vehicle collisions compared to C. thous, D. novemcinctus, N. nasua and D.albiventris. In theory, the number of animal-vehicle collisions should depend on the density and activity of animals and number of vehicles (Seiler & Heldin, 2006). For some species, the low number of roadkill events is mainly related to its low abundance in nature (e.g., the large felines that in general have their population rarefied in an area) (Jaeger et al., 2005). In addition, records of roadkills may reinforce the vulnerability of some species to landscape modifications. Among the 17 species roadkilled on PR-445, four of them (Sapajus nigritus, Leopardus guttulus, Leopardus wiedii, Herpailurus yagouaroundi) are listed in the "Vulnerable" or "Nearly threatened" categories, both on national and international scales (MMA, 2018; IUCN, 2019) and Sylvilagus brasiliensis is listed in the "Vulnerable" category on a regional scale (Paraná, 2010). Although it is expected that species with nocturnal habit are more susceptible to being run over on highways than those of diurnal habit (Bruinderink & Hazebroek 1996), this was not evident in our study. We did not find significant differences in roadkills in relation to species activity patterns. Moreover, peaks in activity pattern are strongly related to climatic conditions and are most evident in groups of amphibians, reptiles and migratory birds (Costa et al., 2015). Our initial hypothesis was that herbivorous and carnivorous mammals were more run over because they tend to walk and cover larger areas than omnivorous and insectivorous animals for food (Tucker et al. 2014). However, our results indicate that omnivorous species are more prone to being run over than herbivorous or carnivorous (figure 2 B). The answer to this pattern may be related to the fact that 55 for the Brazilian Atlantic Forest most mammal species are omnivorous and have a generalist and opportunistic behavior (Reis et al. 2009, Paglia et al. 2012), and even in a region densely fragmented and with small forest remnants surrounded by an agricultural matrix as the region of our study, mammals tend to look for resources in other landscapes outside the forests such as the edges of roads in search of resources such as the remains of food discarded by humans, fruits of edge trees and open area insects. Our results also indicate that the analysis of the cuticular and medullary pattern of mammal hairs is a useful tool for identifying roadkill mammals. This technique allows the confirmation of species identification considering non-intact individuals — particularly small wild felines — because the loss of the main characters utilized in species identification can be destroyed by vehicle collisions. While there are few reports using this tool for roadkill studies, this technique is limited to studies exclusively related to mammals (Quadros & Monteiro-Filho, 2010). In Brazil, there are initiatives such as the monitoring of vehicle collisions with wildlife carried out by the Brazilian Center for Road Ecology Research (CBEE) at the Federal University of Lavras (UFLA), including a monitoring system through citizen science (i.e., Urubu System App) providing information for mitigation measures for animal accidents on Brazilian roads. Other important actions have been developed by the environmental enterprise ViaFauna in São Paulo, called Passa-Bicho (“Animal Pass”). This initiative enables the detection of animals based on positioning a set of sensors on stretches of roadway with high animal traffic (Vasconcelos, 2017; Abra et al., 2019). Still, the responsibility of managing the roads rests on concessionaires, either private or stat, to develop conservation strategies and public policies to reduce road impacts on biodiversity. The roads in Paraná are among those that have the highest roadkill records in Brazil (CBEE, 2019). Our results indicate the sections of higher incidence of animals run over on PR-445 (See Fig. 1). Thus, density analysis based on kernel estimation proves to be an important tool for the planning of actions, where the most likely areas of collisions can be accurately indicated. We emphasize that all along this highway, there are no basic systems to avoid collisions with wild animals. The installation of 56 underpasses and external fences could be suitable mitigation measures for mammals (Rytwinski et al., 2016), especially in regions of the road near bridges crossing over rivers and streams, as well as in regions with higher concentrations of native vegetation. We also suggest placing speed reducers (warning signs, speed cameras and speed bumps) and road fencing, which should be installed mainly in the places where this study indicated the highest roadkill density. We emphasize that the installation of these fences must be in strategic locations along the highway that lead to underground passages. Thus, forcing animals to use them to safely access the other side of the road and reduce the risk of collision for cars. Finally, it is necessary to increase incisive advertising campaigns aimed at drivers, so that is a is way between the road transport authority and the public, it is possible to reduce collisions between vehicles and animals. ACKNOWLEDGMENTS We thank the Postgraduate Program in Biological Sciences of the Universidade Estadual de Londrina and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES- Funding Code 1689817), for logistical and fnancial support. JAB is supported by a postdoctoral fellowship grant 2018- 05970-1, São Paulo Research Foundation (FAPESP). Dr. A. Leyva (USA) provided English editing of the manuscript. REFERENCES Abra, F.D.; Granziera, B.M.; Huijser, M.P.; De Barros Ferraz, K.M.P.M.; Haddad, C.M. & Paolino, R.M. 2019. Pay or prevent? Human safety, costs to society and legal perspectives on animal-vehicle collisions in São Paulo state, Brazil. PLoS ONE, 14: 1-22. Agência Nacional de Transportes Terrestres (ANTT) 2019. Rodovias. Available at: http://www.antt.gov.br/rodovias/index.html http://www.antt.gov.br/rodovias/index.html 57 Araújo, D.R.; Ribeiro, P. & Teles, L.T. 2019. 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Publicatio UEPG: Ciências Biologicas e Da Saúde, 18: 121-133. 63 CAPÍTULO 3 Modeling the geographic distribution of Myocastor coypus (Mammalia, Rodentia) in Brazil: establishing priority areas for monitoring and an alert about the risk of invasion Alan Deivid Pereira, Ricardo, José Ricardo Pires Adelino, Diego Azevedo Zoccal Garcia, Armando Cesar Rodrigues Casimiro, Ana Carolina Vizintim Marques, Ana Paula Vidotto-Magnoni, Sergio Bazilio & Mário Luís Orsi Capítulo redigido e publicado segundo as normas do periódico Studies on Neotropical Fauna and Environment, disponível em: https://doi.org/10.1080/01650521.2019.1707419 https://doi.org/10.1080/01650521.2019.1707419 64 Studies on Neotropical Fauna and Environment ISSN: 0165-0521 (Print) 1744-5140 (Online) Journal homepage: https://www.tandfonline.com/loi/nnfe20 Modeling the geographic distribution of Myocastor coypus (Mammalia, Rodentia) in Brazil: establishing priority areas for monitoring and an alert about the risk of invasion Alan Deivid Pereira, José Ricardo Pires Adelino, Diego Azevedo Zoccal Garcia, Armando Cesar Rodrigues Casimiro, Ana Carolina Vizintim Marques, Ana Paula Vidotto-Magnoni, Sergio Bazilio & Mário Luís Orsi To cite this article: Alan Deivid Pereira, José Ricardo Pires Adelino, Diego Azevedo Zoccal Garcia, Armando Cesar Rodrigues Casimiro, Ana Carolina Vizintim Marques, Ana Paula Vidotto- Magnoni, Sergio Bazilio & Mário Luís Orsi (2020): Modeling the geographic distribution of Myocastor coypus (Mammalia, Rodentia) in Brazil: establishing priority areas for monitoring and an alert about the risk of invasion, Studies on Neotropical Fauna and Environment, DOI: 10.1080/01650521.2019.1707419 To link to this article: https://doi.org/10.1080/01650521.2019.1707419 View supplementary material Published online: 01 Jan 2020. Submit your article to this journal View related articles View Crossmark data Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=nnfe20 https://www.tandfonline.com/loi/nnfe20 https://doi.org/10.1080/01650521.2019.1707419 https://www.tandfonline.com/doi/suppl/10.1080/01650521.2019.1707419 https://www.tandfonline.com/action/authorSubmission?journalCode=nnfe20&show=instructions https://www.tandfonline.com/doi/mlt/10.1080/01650521.2019.1707419 https://www.tandfonline.com/action/journalInformation?journalCode=nnfe20 65 STUDIES ON NEOTROPICAL FAUNA AND ENVIRONMENT https://doi.org/10.1080/01650521.2019.1707419 ORIGINAL ARTICLE Modeling the geographic distribution of Myocastor coypus (Mammalia, Rodentia) in Brazil: establishing priority areas for monitoring and an alert about the risk of invasion Alan Deivid Pereira a, José Ricardo Pires Adelino a, Diego Azevedo Zoccal Garcia b, Armando Cesar Rodrigues Casimiro a, Ana Carolina Vizintim Marques a, Ana Paula Vidotto-Magnoni b, Sergio Bazilio c and Mário Luís Orsi b aPrograma de Pós-Graduação em Ciências Biológicas, Centro de Ciências Biológicas, Departamento de Biologia Animal e Vegetal, Universidade Estadual de Londrina, Londrina, Brazil; bCentro de Ciências Biológicas, Departamento de Biologia Animal e Vegetal, Laboratório de Ecologia de Peixes e Invasões Biológicas, Universidade Estadual de Londrina, Londrina, Brazil; cCampus de União da Vitória, Universidade Estadual do Paraná – UNESPAR, União da Vitória, Brazil ARTICLE HISTORY Received 16 January 2019 Accepted 15 December 2019 KEYWORDS Biological distribution; ecosystem engineering; invasion; risk assessment; nutria Introduction Popularly known as coypu or nutria, Myocastor coypus (Molina 1782) is a large semi-aquatic rodent and the only species comprising the Myocastoridae family (Mammalia, Rodentia) (Woods et al. 1992). Native to South America, typically near the Maipo River in the Santiago province (Chile), the species was originally distributed from Patagonia (a sub-region of northern Argentina) to Bolivia, Paraguay, Uruguay, and Chile, as well as in far southern Brazil (Woods et al. 1992). Myocastor coypus was also introduced to Europe, Asia, Africa, and North America for fur farming and meat production. It is considered a highly invasive species in many countries today (Carter & Leonard 2002). This species can alter natural habitats by feeding on aquatic vegetation, destroying nests, and preying on the eggs of several aquatic birds (Scaravelli 2002). Additionally, it can feed on a variety of crops and weaken riverbanks through its burrowing activity (Carter & Leonard 2002; Guichón & Cassini 2005). Due to the negative impact of this species to the econ- omy, freshwater environments, and agricultural activ- ities (Bertolino 2009), it is ranked as one of the top 100 worst invasive species in the world (Lowe et al. 2000). Several countries carry out permanent population con- trol campaigns (Carter & Leonard 2002; Pepper et al. 2017), with successful eradication undertaken in two small regions of the United States (Carter & Leonard 2002; Jarnevich et al. 2017; Pepper et al. 2017), as well as larger areas within the United Kingdom (Jarnevich et al. 2017). In this context, it is crucial to investigate the relationship between M. coypus and the non-native environments it inhabits in order to anticipate and avoid potential negative environmental and economic impacts. Few ecological studies on this species were per- formed in Brazil, and, to date, its geographical limits and facilitating factors concerning access to new areas © 2019 Informa UK Limited, trading as Taylor & Francis Group Published online 01 Jan 2020 ABSTRACT Myocastor coypus is a large semi-aquatic rodent ranked as one of the 100 most invasive species in the world. In Brazil, few ecological studies have been proposed to understand the relationship of this species with the environment. To date, drivers that facilitate its establishment in new areas remain unknown. However, it is generally accepted that the natural distribution of this species is limited to far southern Brazil. The present study aimed to understand the geographic distribution of M. coypus and indicate areas of greater risk of establishment based on bioclimatic predictors and a surveillance map. We observed that M. coypus suitability and risk assessment areas are restricted to the southeastern and southern regions of Brazil and that anthropogenic landscape modifications are an adequate variable to explain the occurrence of the species. Due to the environmental impacts caused by this species, monitoring in environments where it has been introduced is required. The model used herein presented efficient applicability and fit for Brazil. Preventive actions and the management of M. coypus in predicted regions prior to its establish- ment are recommended. mailto:alandeivid_bio@live.com https://doi.org/10.1080/01650521.2019.1707419 2 A. D. PEREIRA ET AL. 2 remain unknown. Among the few studies that reported the economic or environmental impacts caused by M. coypus in Brazil, Odebrecht et al. (2013) reported that, during the El Niño of 2002 and 2003, M. coypus reduced the biomass of smooth cordgrass (Spartina alterniflora Loisel) by 80% from the low marshes at Lagoa dos Patos in the state of Rio Grande do Sul. In the state of Santa Catarina, M. coypus has been pointed out as responsible for the disruption of fish farming ponds (S. Bazilio, pers. comm., 2018). Due to the limited information about the occurrence of this spe- cies in Brazil, it is difficult both to estimate its relation- ship with the environment and to develop the appropriate management and conservation responses. Therefore, it is necessary to employ statistical tools that can be prioritized for decision-making instead. Environmental niche models (ENMs) are a common tool in ecology, and a practical approach to under- standing potential species distribution based on the relationship between occurrences and environmental information (Franklin & Miller 2010). As ENMs allows for spatial and temporal transferability, a large number of studies use niche models in a biological invasion context (Jiménez-Valverde et al. 2011). The main pur- poses of these models are 1) to anticipate species intro- ductions (Farashi & Najafabadi 2015, 2017; Adelino et al. 2017; Hilts et al. 2019); 2) build risk assessment models (Jiménez-Valverde et al. 2011; West et al. 2018; Hilts et al. 2019), and 3) generate conservation and decision-making information (Venette et al. 2010; Hilts et al. 2019). Thus, ENMs are useful tools to detect areas with higher quality environmental conditions for M. coypus. Therefore, with the knowledge that M. coypus currently occurs in non-native areas (based on IUCN data), we aimed to 1) understand the geographic distribution of M. coypus based on bioclimatic predictors, and 2) create a surveillance map to indicate the areas in which this species is more likely to establish itself once it has access to them. Material and methods Native occurrence data Native species occurrence data (N = 9,563) was obtained from the Global Biodiversity Facility Database (www.gbif.com). Species locality data attached in digital databases are often biased (Maldonado et al. 2015) as sampling is undertaken in easily accessible areas, or within close proximity to roads (Kadmon et al. 2004). Therefore, sampling bias can affect the accuracy of the prediction of environ- ment niche models (Peterson et al. 2008). In order to control the negative effects of spatial sampling biases in environment niche models, we applied a spatial filter analysis (Boria et al. 2014). The spatial filter uses randomization algorithms to optimize the geographical distance between each occurrence record within a required geographical dis- tance. This approach allows for controlling the spatial correlation in the occurrence data and is crucial to reduce the effects of uneven or biased occurrence points for a given species (Aiello- Lammens et al. 2015). Here, we run the spatial filter algorithm imple- mented in the R package (spThin) as proposed by Aiello- Lammens et al. (2015) using a minimum dis- tance parameter of 10 km. Although the minimum distance from each occur- rence point can be controlled according to the species’ movement ability, for our focal species, we understand that a 10 km distance accounts for this (Woods et al. 1992). Therefore, it is useful to ensure the spatial inde- pendence of observed occurrences. In order to max- imize the number of independent spatial occurrences, we repeated the procedure 100 times and used the repetition that presented the greatest number of occur- rences (N = 104). This occurrence data (Native data hereafter) was used to model the environmental niche relating to the M. coypus native occurrence range (Supplementary Material 01). Brazilian occurrence data Species occurrences in Brazil were obtained through the following steps: 1) Searching published data avail- able in scientific journals (N = 17), and 2) Field obser- vation (N = 5) using an ad libitum technique (Altman 1974). The latter was obtained through field incursions in the northern and southern regions of the state of Paraná (PR), and in the northern region of the state of Santa Catarina (SC) between 2012 and 2017. In order to formally record the presence of the species in a new area, one specimen sampled in Londrina (PR) was deposited in the Museum of Zoology of the Universidade Estadual de Londrina, Londrina, PR (Voucher Number MZUEL 376). The searches for available scientific articles were performed using the advanced research tool in Web of Science, Scopus, Google Scholar, and Scielo databases. For each one, we used the following keyword combinations: i) ‘Myocastor coypus’ and ‘Brazil,’ ‘Ratão do banhado’ and ‘Brazil,’ ‘Myocastor coypus’ and ‘Brazilian Territory,’ and ‘Myocastor coypus’ and ‘occurrence.’ For all useful articles, we collected the geographical coordinates, locality (municipality), state, and biome (Supplementary Material 02). These occurrences http://www.gbif.com/ 3 (Brazil data hereafter) were used to model the environ- mental niche of M. coypus occurrence range in Brazil. Environmental data and variable selection Climate data were obtained from the WorldClim database version 2.0 (Fick & Hijmans 2017). We use Principal Component Analysis (PCA) to select, reduce and producing uncorrelated variables (Legendre & Legendre 1998; Peterson et al. 2011; Adelino et al. 2017). This was done by keeping the variable with the highest loading when variables were correlated (Supplementary Material 03), considering variables that are relevant to the ecology of the M. coypus. The procedure was performed inde- pendently by each dataset (i.e. Native and Brazil data set). Additionally, because of the presence of M. coypus in human-associated landscapes, we chose to input the human footprint environmental layer as additional environmental information (Wildlife Conservation Society, Center for International Earth Science Information Network, 2005; available at http://sedac. ciesin.columbia) for measuring the Human Influence Index (HII) in the landscapes. Environmental niche models (ENM) Because of the differences in the approaches to model- ing the species environmental niche (Franklin & Miller 2010), a combination of different algorithms in the form of an ensemble model should be employed (Araújo & New 2007). Ensemble models are used as a means of combining different algorithms in a weighted approach. Furthermore, because niche models can be grouped by the level of complexity (Rangel & Loyola 2012) and type of data used for the model, we used three different classes of algorithms (i.e. confidence interval, distance measure, and machine learning) to model the potential distribution of M. coypus. We used the following environmental niche mod- els: Bioclim (Nix 1986; Booth et al. 2014), Domain (Carpenter et al. 1993), Support Vector Machine (Guo et al. 2005; Drake et al. 2006), and Maxent (Phillips et al. 2004). We chose these distribution models due to their capacity for model species- environment relationships using presence-only data (Franklin & Miller 2010) In order to generate the information modeled by each dataset, we combined the native-only and the Brazilian- only models into one final model (i.e. ensemble). The ensemble approach helps to reduce the dissimilarity between the models and enables an improvement in the predictive capacity of the final model (Araújo & New 2007). For each dataset, we ran 100 repetitions by each algorithm and used the mean suitability values overall models to generate the final suitability model (see Supplementary Material 04 for more details). Model calibration and model evaluation For each model, we used 75% of the occurrence data (Nnative = 78, NBrazil = 16) to calibrate the models, and 25% of the occurrence data (Nnative = 26, NBrazil = 6) to validate the models. Because we had presence-only information, we used the pseudo absence technique to simulate the absence data in the geographical space. Pseudo absence randomly samples the cells within the geographical region to create background information, which is then used to test the accuracy of the model (Franklin & Miller 2010). The most traditional measure of accuracy in niche models is the Area Under the Receiver Operating Characteristic Curve (ROC), which measures the capacity of the model to discriminate between true positive values and false positives (Fielding & Bell 1997). However, this measure can be overestimated (Peterson et al. 2008), and different approaches can be used to measure the accuracy of the model (Allouche et al. 2006; Peterson et al. 2008). In order to reduce the evaluation biases, we evalu- ated the model fit using the partial area under the ROC curve (Peterson et al. 2008). This was performed in the ‘Partial ROC’ software (Barve 2008). We performed the Partial ROC approach using 1,000 iterations. For each one, 50% of the test data (i.e. bootstrap) was re- sampled, and a 95% error was accepted. Models that had partial pAUC ratio >1 were considered as being better than a random performance and were combined in one final model (Peterson et al. 2008). Surveillance map To estimate the potential distribution of the species in Brazil, we used the sensitivity = specificity threshold. This allowed us to identify all the grid cells that can be estimated as presence (cell value = 1) or absence (cell value = 0) for the species (a.k.a. Binary models) in each algorithm and dataset with pAUC ratio >1. We chose the sensitivity = specificity threshold because it is an acceptable measure that minimizes the mean positive and negative error rates (Liu et al. 2005). This procedure produced eight binary maps (4 algo- rithms x 2 regions). All binary models were then over- lapped, and the values of each grid cell were summed http://sedac.ciesin.columbia/ http://sedac.ciesin.columbia/ 4 A. D. PEREIRA ET AL. 4 to create a surveillance map. The surveillance map represents the agreement of the models to predict each grid cell as a potential region for the occurrence of the species. This approach was previously suggested to increase model accuracy by reducing the variability of projected models (Araújo & New 2007) and has been previously applied in different ecological situations (Carnaval & Moritz 2008; Porto et al. 2013). Considering the context of invasive species stu- dies, if all models predict the same grid cell as suitable for the presence of the species, then the grid cell represents an important geographical region for surveillance. Conversely, if all models predict the same grid cell as unsuitable for species presence (i.e. absence), then the grid cell represents a less impor- tant geographical region for surveillance. We then used the quantiles of the summed values of each grid cell (Nsum = 8) to assign one of four classi- fications for the Surveillance index: 1) Without Surveillance: less than two models predict the cell as an occurrence; 2) Lower Surveillance: two to four models predict the cells with presence; 3) Moderate Surveillance: four to six models predict the cells with presence, and 4) Great Surveillance: more than six models predict the cells with presence. All analyzes were conducted using the R software (R Core Team 2015) dismo package (Hijmans et al. 2016). Results The literature review resulted in 17 published studies between 2000 and 2016 (Table S1) and included specimens from five states covering the Southern and Southeast regions of Brazil and throughout two biomes – Pampa and Atlantic Rainforest (Figure 1). Myocastor coypus was recorded in five new locations from 2012 to 2017, four in the state of Paraná and one in the state of Santa Catarina. All algorithms were considered as presenting a good model fit (Table 1). Variables presenting the highest relative importance values were ‘Human Footprint,’ ‘Mean Temperature of the Coldest Quarter,’ and ‘Minimum Temperature of the Coldest Month’ (Table 1). Human Footprint was positively associated with M. coypus suitability (r = 0.6), while Mean Temperature of the Coldest Quarter (r = −0.48), Min Temperature of the Coldest Month (r = −0.35), and Annual Precipitation (r = −0.31) Figure 1. Mycastor coypus records in Brazilian biomes and the suitability values for each record. 5 Table 1. Partial area under the curve (pAUC) and threshold values (sensitivity = specificity) for the four algorithms used to model the potential distribution of Myocastor coypus (models were combined if pAUC > 1). The relative importance of each variable used to model the distribution of Myocastor coypus are displayed below the table. Native data Brazil records Model metrics pAUC ratio Threshold pAUC ratio Threshold Bioclim 1.745 ± 0.211 0.09636443 1.966 ± 0.000 0.1558118 Domain 1.619 ± 0.478 0.64056412 1.997 ± 0.000 0.4945468 Maxent 1.767 ± 0.283 0.34659937 1.994 ± 0.000 0.4151913 SVM 1.731 ± 0.341 0.48329812 1.999 ± 0.000 0.5336144 Combined model Environmental layers Relative importance Human Footprint 0.333 Mean Temperature of Coldest Quarter 0.208 Min Temperature of Coldest Month 0.201 Temperature Annual Range 0.116 Precipitation of Wettest Quarter 0.091 Annual Precipitation 0.051 were negatively associated with the presence of this species (see Supplementary Material 05). Suitable regions were concentrated along the south- eastern and southern regions of Brazil (Figure 2a). Areas exhibiting greater suitability values for the occur- rence of the species are concentrated in the eastern and western mesoregions of the state of Rio Grande do Sul, the southern region of the state of Santa Catarina, and Figure 2. Suitability map for Myocastor coypus occurrences in Brazil (a). South and Southeast Brazil, with confirmed occurrences in the published scientific literature (b). Low suitability values are represented by green and high suitability values are represented by red. Highlighted, a photographic record of M. coypus through opportunistic observations in northern of the state of Santa Catarina and the record of a run-over specimen on the PR-538 highway, near the Cafezal River, in the municipality of Londrina, PR. 6 A. D. PEREIRA ET AL. 6 the south and southeastern regions of the state of Paraná (Figure 2b). The highest suitability values were observed for the Pampa biome at 0.63, followed by the Atlantic Rainforest Biome, at 0.59. The lowest suitabil- ity value was found for the ecotone regions between the Atlantic rainforest and the Cerrado, at 0.24 (Figure 1 and Table S1). The surveillance map (i.e. the combination of binary maps) (Figure 3a; for more detail see Supplementary Material 04) indicated that the pre- valence observed for the South and Southern regions were classified at lower and moderate grid cell values. High-risk classification cells were con- centrated in the states of Santa Catarina and the Rio Grande do Sul. Further, small high-risk patches were distributed along with the state of Paraná (Figure 3b). Of all 22 occurrences in Brazil, 50% (N = 11) fell into grid cells classified as moderate risk, 22.7% (N = 5) as lower risk, 13.6% (N = 3) as without risk, and 9.09% (N = 2) as high risk (Figure 3c). Discussion This study was the first to compile information about the current occurrence of M. coypus in the Brazilian territory, as well as modeling the occur- rence of this species with a more accurate and updated database based on confirmed occurrences. Here, we considered this species to be native to the extreme southern Brazil only (Pampa biome), diver- ging from the information provided by IUCN but in accordance with studies of the species’ natural his- tory, which proposed that southern part of the state Figure 3. Surveillance map, combined binary map for the occurrence of Myocastor coypus in Brazil. (a) The southeastern and southern regions of Brazil are highlighted. (b) Record percentages in relation to risk assessment for Brazil areas. B 7 of Rio Grande do Sul could be considered its native region (Woods et al. 1992). Our review of the occurrence points of M. coypus for Brazil indicated that its presence in southeastern Brazil is attributed to escapes from fur farms, meat production, and deliberate introductions in lagoons and rivers (Bueno 2013). Currently, this species is easily regis- tered in flooded pastures and floodplain areas in the states of São Paulo and Rio de Janeiro (non-native region) (Rocha et al. 2004; Bueno 2013). In the state of Paraná, this species is already on the list of invasive alien species, and its transportation, crea- tion, release or translocation, cultivation, propaga- tion (by any means of reproduction), commercialization, donation, or intentional acquisi- tion in any form is prohibited (IAP 2015). The suitability model developed in the present study suggested suitable areas in Brazil were mainly found in the Atlantic Forest biome in the states of Santa Catarina, Paraná, São Paulo, and Rio de Janeiro (i.e. the Southeastern region of Brazil) (Figure 2). Our models identified that some environmental variables, such as variations in temperature, as well as the annual mean precipitation, explain the current range of M. coypus. We found that Annual Precipitation and Precipitation of Driest Quarter are important climatic contributors in predicting suitable areas for the M. coypus. These climatic variables had a negative effect on suitability of coypu, which indicate that this species can be favored in dry regions provided that they are associated with water resources (e.g. rivers and streams) as shown in previous studies (Carter & Leonard 2002; Hong et al. 2014; Farashi & Najafabadi 2015, 2017; Hilts et al. 2019). Regarding temperature variables, according to the models developed by Gosling et al. (1983) and Jarnevich et al. (2017), M. coypus suffers population declines due to the adverse temperature effects in environments with temperature ranging between zero and five degrees (i.e. freezing days). However, even in its native range (i.e. Patagonia Region), this species is expected to survive low temperatures during severe winters, provided that ice sheets do not form over waterbodies for long periods of time (Ehrich 1962). Previous studies have reported that nutria densities declined 71% after river channels were frozen for 20 days (Doncaster & Micol 1989). Recently, Hilts et al. (2019) modeled the distribution of M. coypus based on climate change projections for 2050. As a result, the authors found that, on a broad scale, areas with ≤80 annual freezing days are more suitable for M. coypus establishment and prosperity. At local scales, habitat covariates, (e.g. areas with high proportions of freshwater forested-shrub wetlands close to other wet- lands) better reflect the ecological niche of this species (Hilts et al. 2019). In this context, for Neotropical regions where there are no sequential freezing days (such as Brazil), the temperature may not be a limiting factor for the occurrence of M. coypus. We believe that the establishment of this species in new areas is dependent only on access to new regions as well as the availability of resources in these areas. The Human Footprint variable (i.e. landscape modification by human activities) was the most important predictor related to the occurrence of M. coypus in Brazil. Other researchers indicated that human disturbance represented by human popu- lation density in urban areas, distance to roads, dis- tance to settlements and urban areas, and bare and rocky areas are important factors affecting the M. coypus distribution (Guichón & Cassini 2005; Bertolino & Ingegno 2009; Farashi & Najafabadi 2015). This positive correlation may be an indicator of the species’ resilience in human environments (i.e. synanthropic species), a hitherto underestimated attribute for this species. Due to widespread damage caused to ecosystems, with effects on crops and riverine vegetation by M. coypus (Vilà et al. 2010), several control and eradication programs have been carried out in countries where the species was introduced (Carter & Leonard 2002; Pepper et al. 2017). This species is capable of weakening dikes and irrigation structures through the grazing and undermining of riverbanks by burrowing (Sofia et al. 2017), and can, therefore, be considered an environmental engineer. According to Sofia et al. (2017), the most critical damage caused by M. coypus – in purely economic terms – is related to drainage structures, with direct costs for the structure management, and indirect costs connected to flooding. In recent years, Coypu- related problems have been increasing rapidly in several countries (Carter & Leonard 2002; Pepper et al. 2017). However, there is still no quantified information about the economic and environmental impact caused by this species in Brazil. The surveillance map indicated that the Southeastern and Southern regions of Brazil could be considered more vulnerable to the establishment of M. coypus. Larger patches of the Santa Catarina coast and of the Rio Grande do Sul mainland were pointed out as the areas with the highest invasion risk, provided this species has access to them (see Figure 3). This could be related to the latitudinal simi- larity; therefore, the environmental congruence between the north region of the species’ native range 8 A. D. PEREIRA ET AL. 8 and the Brazilian observed range. Conversely, the data presented herein suggest stronger environmental dis- similarity between the North region of the species’ native range and Southeast of the detected Brazilian range. This can be observed by the prevalence of lower invasion risk classes in the states of São Paulo, Rio de Janeiro, Espírito Santo, and Minas Gerais. Indeed, because climate matching is an important assumption for reproduction success between environments (Holt et al. 2005). The dispersion access of this species to new areas may easily occur in lower Brazilian latitudes, as human landscape modifications and river fragmen- tation by dams may facilitate its establishment. Such dams may also exhibit greater aquatic macrophyte abundances, which are an important primary food requirement for M. coypus (Guichón et al. 2003). We also found strong indicators that it was intro- duced in the northern and southern regions of Paraná in addition to documented evidence that it was introduced to the Southeast of Brazil. For this reason, further studies on the possible introduction in new areas and subsequent outcomes for M. coypus are essential for management mea- sures. The model used herein presented efficient applicability and fit and can also be extended to other non-native invasive species. Our results may be an adequate basis for further studies aimed at understanding the dispersion and distribution of M. coypus in Brazil, as well as for management actions, if necessary. Acknowledgments We thank the Postgraduate Program in Biological Sciences of the Universidade Estadual de Londrina and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES- Funding Code 1689817), for logistical and financial support. Disclosure statement No potential conflict of interest was reported by the authors. Funding This work was supported in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - [Proc.1689817];Coordenação de Aperfeiçoamento de Pessoal de Nível Superior [1689817]. ORCID Alan Deivid Pereira http://orcid.org/0000-0002-3182- 2344 José Ricardo Pires Adelino http://orcid.org/0000-0002- 8637-2838 Diego Azevedo Zoccal Garcia http://orcid.org/0000- 0001-5709-6347 Armando Cesar Rodrigues Casimiro http://orcid.org/ 0000-0001-8826-5609 Ana Carolina Vizintim Marques http://orcid.org/0000- 0003-4978-9260 Ana Paula Vidotto-Magnoni http://orcid.org/0000-0003- 1819-7019 Sergio Bazilio http://orcid.org/0000-0003-3577-8931 Mário Luís Orsi http://orcid.org/0000-0001-9545-4985 References Adelino JRP, Anjos L, Lima MR. 2017. Invasive potential of the pied crow (Corvus albus) in eastern Brazil: best to eradicate before it spreads. Perspect Ecol Conserv. 15 (3):227–233. 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Woods CA, Contreras L, Willner-Chapman G, Whidden HP. 1992. Myocastor coypus. Mamm Species. 398:1– 8. https://doi.org/10.1890/080083 74 A. D. PEREIRA ET AL. 74 SUPPORT MATERIAL 75 Support Material Modeling the geographic distribution of Myocastor coypus (Mammalia, Rodentia) in Brazil: Establishing priority areas for monitoring and an alert about the risk of invasion. Alan Deivid Pereiraa*, José Ricardo Pires Adelinoa, Diego Azevedo Zoccal Garciab, Armando Cesar Rodrigues Casimiroa, Ana Carolina Vizintim Marquesa, Ana Paula Vidotto-Magnonib, Sergio Bazilioc, Mário Luís Orsib 1. Spatial filter Figure. S1. Occurrence points before and after spatial filter. 2. Occurrence data Table S1. Myocastor coypus coordinates occurrence records in Brazil. 3. Selection of Bioclimatic variables Table S2: PCA loading values of GBIF thinned data set inside native range. Table S3: PCA loading values from Brazilian thinned data set. Figure. S2: Ordination plot of climate variables. All results were obtained only with native data according with IUCN. Figure. S3: Ordination plot of climate variables. Loading values results with native and Brazil occurrences 4. Ecological Niche Models and model evaluation Figure. S4: Suitability models calibrated with GBIF native occurrences and projected within Brazilian range. Figure. S5: Suitability models calibrated with Brazilian occurrences (i.e. data set two) and projected within Brazilian range. Figure. S6. Binary classification maps for the four different niche models used to model M. coypus. Each classification maps were obtained from GBIF occurrences in native range and projected within Brazilian range. Figure. S7. Binary classification maps for the four different niche models used to model M.coypus. 5. Supplementary Results Figure. S8. Correlation plot. 76 A. D. PEREIRA ET AL. 76 Fig. S1. Occurrence points before and after spatial filter. Blue bar = Raw GBIF coordinate data points before spatial filter, Purple = Thinned GBIF coordinate data points after spatial filter, Red = Raw Brazil coordinate data points before spatial filter, Green = Thinned Brazil coordinate data points before spatial filter. 1. Spatial filter 77 Table S1. Myocastor coypus coordinates occurrence records in Brazil. Locality: Municipality in which each species coordinate was observed, State: State in which each species coordinate was observed: Rio Grande do Sul (RS); Santa Catarina (SC); Paraná (PR); São Paulo (SP); Rio de Janeiro (RJ), Biome: landscape classification in which each species coordinate was observed, Reference: information source in which each species coordinate was gathered, Map ID: point label for each occurrence in map reference (see Fig. 2 in manuscript), NR indicate ad libitum observations by S. Bazilio or M. L. Orsi and DR indicate dead species founded in the road .Suitability: suitability values for each occurrence points in Brazil. Latitude Longitude Locality State Biome Reference Map ID Suitability 30°4’8.97”S 51°1’2.70”W Viamão RS Pampa Cademartori & Machado, 2002 1 0.4124746 30°11’29”S 52°21’25”W Pantano Grande RS Steil et al., 2016 2 0.5056855 29°42’59.11”S 53°42’42.25”W Santa Maria RS Santos et al., 2008 3 0.6287537 31°15’17.40”S 50°58’18.01”W Tavares RS Colares et al., 2000 4 0.2876192 29°49’30.00”S 54°49’30.00”W Cacequi RS Pinto & Duarte, 2013 5 0.4985294 22°52’48.85”S 47°0’54.12”W Campinas SP Cerrado/ Atlantic Rain Forest Angelo et al., 2016 6 0.2057709 21°56’01”S 42°36’31”W Rio de Janeiro RJ Atlantic Rain Forest Rocha et al., 2004 7 0.3532902 22°07’09”S 43°09’22”W Três Rios RJ Bueno, 2013 8 0.5174042 22°44’21”S 46°54’27”W Pedreira SP Lemos et al., 2004 9 0.4967758 23°31’22”S 46°11’18”W Mogi das Cruzes SP Martins et al., 2017 10 0.4229938 25°26’4.59”S 49°3’18.90”W Piraquara PR Cáceres, 2004 11 0.4773234 26°06’37”S 53°33’40”W Bom Jesus do Sul PR Wolfart et al., 2013 12 0.2595832 24°42’49”S 53°44’35”W Toledo PR Junior & Silva, 2015 13 0.3648623 25°26’59”S 52°54’29”W Quedas do Iguaçu PR Juraszek et al., 2014 14 0.570746 24°34’22”S 49°55’35”W Piraí do Sul PR Grazzini et al., 2015 15 0.3089292 27°16’00”S 50°26’26”W São Cristóvão do Sul SC Cherem et al., 2007 16 0.5239566 26°42'44.98"S 49°28’51.39”W Doutor Pedrinho SC Tortato et al., 2014 17 0.4816769 23°19’37.12”S 51°10’32.54”W Londrina PR Present study (DR) 18 0.4839047 24°57’21”S 53°27’19"W Cascavel PR Present study (NR) 19 0.2511306 26°13’48”S 51°05’11”W União da Vitória PR Present study (NR) 20 0.5668645 26°29’03”S 51°59’26”W Palmas PR Present study (NR) 21 0.4234352 26°06’23”S 50°19’20” W Três Barras SC Present study (NR) 22 0.5867548 2. Occurrence data 78 A. D. PEREIRA ET AL. 78 3. Selection of Bioclimatic variables Climate data were obtained from the WorldClim database version 2.0 (Fick and Hijmans, 2017). The WorldClim database has 19 bioclimatic variables. Because few number of studies focused in species environmental relationship we choose to no previously select the variables used in the model. However, because the negative effects of collinearity variables in environmental niche models we selected the variables using multivariate approach. To do this, we extracted the environmental values from each thinned data set (850 occurrences for the dataset one and 22 for the dataset two) and submit both datasets to a PCA analyses. We used highest loading values for the two principal components to identify correlated variables (i.e., had similar vector directions) (Table S2 and table S3). Table S2: PCA loading values of GBIF thinned data set inside native range. In bold, selected bioclimatic variables names and values of retained variables. PC1 = first component of PCA, PC2 = second component of PCA. Variable Bioclimatic Variable Names PC1 PC2 bio_01 Temperature Annual Range 0.346 0.145 bio_02 Mean Diurnal Range (mean of monthly (max temp – min temp) 0.246 -0.201 bio_03 Isothermality ((BIO2/BIO7)* 100) 0.215 0.128 bio_04 Temperature Seasonality (standard deviation *100) 0.038 -0.284 bio_05 Max Temperature of Warmest Month 0.361 0.021 bio_06 Min Temperature of Coldest Month 0.277 0.252 bio_07 Temperature Annual Range 0.117 -0.305 bio_08 Mean Temperature of Wettest Quarter 0.334 0.076 bio_09 Mean Temperature of Driest Quarter 0.092 0.187 bio_10 Mean Temperature of Warmest Quarter 0.357 0.075 bio_11 Mean Temperature of Coldest Quarter 0.315 0.202 bio_12 Annual Precipitation -0.114 0.36 bio_13 Precipitation of Wettest Month -0.175 0.296 bio_14 Precipitation of Driest Month -0.07 0.276 bio_15 Precipitation Seasonality (coefficient of variation) -0.03 -0.129 bio_16 Precipitation of Wettest Quarter -0.185 0.295 bio_17 Precipitation of Driest Quarter -0.071 0.291 bio_18 Precipitation of Warmest Quarter 0.161 0.287 bio_19 Precipitation of Coldest Quarter -0.302 0.181 79 Table S3: PCA loading values from Brazilian thinned data set. In bold, loading bioclimatic variables names and values of retained variables. PC1 = first component of PCA, PC2 = second component of PCA. The vector directions (Fig S2 and Fig S3) indicate the correlation of variable with the component axes and therefore allow us to choose the variable that more contribute with the variable explanation from each axis. Variable Bioclimatic Variable Names PC1 PC2 bio_01 Temperature Annual Range 0.211 -0.309 bio_02 Mean Diurnal Range (mean of monthly (max temp – min temp) 0.034 0.262 bio_03 Isothermality ((BIO2/BIO7)* 100) 0.215 0.283 bio_04 Temperature Seasonality (standard deviation *100) -0.272 -0.205 bio_05 Max Temperature of Warmest Month 0.145 -0.292 bio_06 Min Temperature of Coldest Month 0.253 -0.261 bio_07 Temperature Annual Range -0.181 0.044 bio_08 Mean Temperature of Wettest Quarter 0.220 0.014 bio_09 Mean Temperature of Driest Quarter 0.054 -0.414 bio_10 Mean Temperature of Warmest Quarter 0.098 -0.396 bio_11 Mean Temperature of Coldest Quarter 0.276 -0.206 bio_12 Annual Precipitation -0.164 0.187 bio_13 Precipitation of Wettest Month 0.259 0.208 bio_14 Precipitation of Driest Month -0.299 -0.041 bio_15 Precipitation Seasonality (coefficient of variation) 0.309 0.075 bio_16 Precipitation of Wettest Quarter 0.267 0.195 bio_17 Precipitation of Driest Quarter -0.298 0.017 bio_18 Precipitation of Warmest Quarter 0.225 0.250 bio_19 Precipitation of Coldest Quarter -0.299 -0.032 80 A. D. PEREIRA ET AL. 80 Fig. S2: Ordination plot of climate variables. All results were obtained only with native data according with IUCN. 81 Fig. S3: Ordination plot of climate variables. Loading values results with native and Brazil occurrences. 82 A. D. PEREIRA ET AL. 82 4. Ecological Niche Models and model evaluation Environmental niche models are computational approach to execute a sequence of mathematical functions that allows correlate species occurrence with environmental data (here called algorithm). This results in the suitability environmental maps which that can be projected among space and time (Peterson, 2003). However, different algorithms result in different ecological dissimilarities and therefore influencing the precision, accuracy and reality of the models (Rangel & Loyola, 2012). Further, a large number of algorithms can be used to model species environmental relationship and are classified in: 1) Distance measures model, 2) Regression methods models and 3) Machine learning methods (Franklin, 2010). Hence, because each environmental niche models are fitted with different mathematical parameters, usually different algorithms result in different predictive models. Although, due to higher variation in model performance (Diniz–Filho et al., 2009), combine multiple models (i.e. ensemble approach) is pointed out as a better alternative to reduce methodological dissimilarity between (Araújo and New, 2007). Therefore, the ensemble approach increases the predictivity capacity of the final model. In this study, we analyzed M.coypus occurrence with the following environmental niche models: Bioclim (Nix, 1986; Booth et al., 2014), Domain (Carpenter et al., 1993), Support Vector Machine (Guo et al., 2005; Drake et al., 2006), and Maxent (Phillips et al., 2004). We choose these distribution models due to the capacity to model species environment relation with presence only data (Franklin, 2010). Each environmental niche model was built using 100 iterations and the mean suitability values were used to generate a final distribution model for each of the four algorithms (Fig S4 and S5). All models presented mean pROC values larger than one. Hence, we combined all models to produce a single final model. To do this, we used the mean suitability per grid cell among all environmental niche models. We then, used the sensitivity = specificity threshold to create a binary map for each data set, resulting in eight predictive models (i.e. four calibrated from Native range projected to Brazil and four calibrated in Brazilian range and projected to Brazil) (Fig.S6 and Fig S7). As a final step, we overlapped all binary maps by summing the values in each grid cell, which resulted in four possible values for the predicted presence. This allows to classify the cells in four classes of risk assessment: (i) Without, when the maximum of two models predict the cell as occurrence; (ii) Lower, when more than two to four models predict the cells with presence; (iii) Moderate, when more than four to six models predict the cells with presence; (iv) Great, when more than six models predict the cells with presence. 83 Fig. S4: Suitability models calibrated with GBIF native occurrences and projected within Brazilian range. Color gradient shows suitability values from blue (i.e. low suitability) to yellow (i.e. Highest values). 84 A. D. PEREIRA ET AL. 84 Fig. S5: Suitability models calibrated with Brazilian occurrences (i.e. data set two) and projected within Brazilian range. Color gradient shows suitability values from blue (i.e. low suitability) to yellow (i.e. Highest values). 85 Fig S6. Binary classification maps for the four different niche models used to model M. coypus. Each classification maps were obtained from GBIF occurrences in native range and projected within Brazilian range. Binary models use threshold values (i.e. here sensitivity = specificity) to classify grid cells in presence and absence class. Grey color indicate cell predicted as absence and red indicate cell predicted as presence. 86 A. D. PEREIRA ET AL. 86 Fig S7. Binary classification maps for the four different niche models used to model M.coypus. Each classification maps were obtained from Brazilian occurrences and projected within Brazilian range. Binary models use threshold values (i.e. here sensitivity = specificity) to classify grid cells in presence and absence class. Grey color indicate cell predicted as absence and red indicate cell predicted as presence. 87 5. Supplementary results Fig. S8. Pairwise correlation values of suitability values obtained by the ensemble approach and the environmental variables used in the modeling procedures for Myocastor coypus in Brazil. 88 A. 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Biote 26, 111–119. https://doi.org/10.5007/2175-7925.2013v26n4p111 https://doi.org/10.4322/natcon.2012.030 http://dx.doi.org/10.17058/cp.v28i1.7855 http://dx.doi.org/10.17058/cp.v28i1.7855 https://doi.org/10.5007/2175-7925.2014v27n3p123 https://doi.org/10.5007/2175-7925.2013v26n4p111 91 CAPÍTULO 4 Mammalian defaunation in Devonian kniferidges and meridional plateaus of the Brazilian Atlantic Rainforest Alan Deivid Pereira, Juliano André Bogoni, Sergio Bazilio, Mário Luís Orsi Capítulo redigido segundo as normas do periódico Biological Conservation, disponível em: https://www.elsevier.com/journals/biological-conservation/0006-3207/guide-for-authors https://www.elsevier.com/journals/biological-conservation/0006-3207/guide-for-authors 92 Mammalian defaunation in Devonian kniferidges and meridional plateaus of the 1 Brazilian Atlantic Rainforest 2 3 Alan Deivid Pereiraª; Juliano André Bogonib,c; Sergio Baziliod; Mário Luís Orsie 4 5 a Programa de Pós-Graduação em Ciências Biológicas, Centro de Ciências Biológicas, 6 Departamento de Biologia Animal e Vegetal, Universidade Estadual de Londrina, 7 Londrina, PR, Brazil, Rodovia Celso Garcia Cid, PR 445, Km 380, CEP 86.057-970 – 8 Londrina, Paraná, Brazil. 9 b Universidade de São Paulo, Escola Superior de Agricultura “Luiz de Queiroz”, 10 Laboratório de Ecologia, Manejo e Conservação de Fauna Silvestre (LEMaC), 11 Piracicaba, São Paulo, Brazil. 12 c School of Environmental Sciences, University of East Anglia, Norwich NR4 7TJ, 13 Norwich, United Kingdom. 14 d Universidade Estadual do Paraná – UNESPAR, Campus de União da Vitória. Caixa 15 Postal 241, CEP 84600‑970, União da Vitória, PR, Brazil. 16 e Laboratório de Ecologia de Peixes e Invasões Biológicas. Universidade Estadual de 17 Londrina, Centro de Ciências Biológicas, Departamento de Biologia Animal e Vegetal, 18 Rodovia Celso Garcia Cid, PR 445, Km 380, CEP 86.057-970 – Londrina, Paraná, 19 Brazil. 20 21 Corresponding author: alandeivid_bio@live.com 22 23 24 25 ABSTRACT 26 Defaunation of mammals may be triggered by commercial hunting, habitat degradation, 27 or the synergic combination of both. Although the population decrease of the mammal 28 community in the Brazilian Atlantic Rainforest has been discussed in past studies, there 29 is scanty information on the southern section of the biome about the defaunation level of 30 remnant forest fragments. The current paper evaluates habitat conversion regarding the 31 decrease of medium and large mammals of the South Atlantic Rainforest through a 32 defaunation index. Results showed a high defaunation degree for the southern Brazilian 33 Atlantic Rainforest, with more than 50% of analyzed fragments revealing higher 34 medium defaunation rates than expected medium ones. Areas with a higher percentage 35 of soil cover for silviculture and agriculture have greater defaunation rates when 36 compared to areas with natural forest cover. Further, protected areas proved to be more 37 balanced functionally and less defaunized than non-protected ones. Big herbivores and 38 carnivores lie within the more defaunation-affected functional categories and loss may 39 cause unstable functional fragments with low resilience rates. 40 41 Keywords: Deforestation; forest fragmentation; land use; mammalian communities; 42 protected areas; tropical rainforest. 43 44 45 46 47 48 49 50 93 INTRODUCTION 51 52 Forest fragmentation, conversion of habitat and overhunting are the main causes 53 for global biodiversity decrease (Fahrig, 2003; Gardner et al., 2009; Wilson et al., 54 2016). Deforestation and habitat conversion are historically more pronounced in littoral 55 regions and the interior plateaus of the Atlantic Rainforest (Dean, 1996; Ribeiro et al., 56 2009), featuring one of the 25 hotspots in world biodiversity (Myers et al., 2000). The 57 Atlantic Rainforest biome has a relevant good percentage of mammal diversity within 58 South America, comprising 321 species, of which 89 (27.7%) are endemic (Paglia et al. 59 2012; Graipel et al., 2017). 60 Due to fragmentation and drastic decrease in the forest coverage in the Atlantic 61 Rainforest, mammal assemblages live in native forest remnants smaller than 100 hectares 62 and immersed within anthropogenic matrixes (Ribeiro et al., 2009). Mammal populations 63 in tropical forests have the highest decrease rates worldwide (Dirzo et al., 2014; Bogoni 64 et al. 2018), with chronic and repeated defaunation events (i.e., local extinctions) (Canale 65 et al., 2012). Defaunation may occur directly by subsistence and commercial hunting and 66 indirectly by fauna decrease due to human activities not specifically aimed at the animals, 67 such as habitat destruction, extractive activities, soil, and water pollution (Redford, 1992). 68 Defaunation may be defined as the decrease in abundance and occurrence of 69 animals in a given community (Terborgh 1988; Peres 1990; Redford 1992), and which 70 may affect in a disproportional way bigger animal with low reproduction rates, 71 especially large mammals (Cardillo et al., 2005). The negative effects of local mammal 72 defaunation may occur at different ecological scales and affect seed dispersal and 73 predation (Stoner et al., 2007; Markl et al., 2012), with the substitution of large 74 predators by medium-sized ones (Taylor et al., 2016) and cause the decrease of other 75 taxonomic groups (Galetti and Dirzo, 2013; Kurten, 2013; Bogoni et al., 2019). 76 It has been calculated that 96% of the Atlantic Rainforest biome is subjected to 77 the effects of trophic cascades due to defaunation of mammals (Jorge et al., 2013). The 78 main defaunation boosters in the Atlantic Rainforest include a long and persistent history 79 of overhunting, conversion, and fragmentation of habitats, or a synergic combination of 80 both (Bogoni et al., 2018). Therefore, the establishment of protected areas (PAs) is one 81 of the most relevant policies run by the government for the preservation of biodiversity 82 (Jenkins and Joppa, 2009). 83 The Devonian Escarpment´s Environmental Protection Area is one of the most 84 relevant regions for conservation in southern Brazil (Ribeiro et al 2009). With over 324 85 thousand hectares, it spreads over a great part of the Campos Gerais in Southern Brazil 86 and forms a specific landscape that alternates pine tree forests, savannas, and rocky 87 outcrops. It is a transition zone between Mixed Ombrophilous Forest, Savanna, and 88 localized Natural Fields (Maack, 2012), and some sites are highly valued for touristic 89 visitation, such as canyons, and archeological and pre-historical ruins (Takeda et al., 90 2001). However, the conversion of natural areas in monocultures, mainly Eucalyptus 91 spp. and Pinus spp. together with agriculture areas threaten the stability of the region´s 92 biodiversity (Almeida and Moro, 2007). 93 Studies on the Brazilian Atlantic Rainforest, at regional scales, have shown high 94 levels of defaunation in mammal assemblages (Galetti et al., 2006; Canalle et al., 2012; 95 Bogoni et al., 2016; Galetti et al., 2017). Most studies on defaunation in the Atlantic 96 Rainforest are concentrated in South-Eastern and Northeastern Brazil (e.g., Galetti et al., 97 2006, 2017; Canale et al., 2012). A biome-scale study, however, showed a larger 98 defaunation in the biome but remains several spatial gaps in terms of defaunation in the 99 high-modified plateaus of the Atlantic Rainforest only accessed via spatial interpolation 100 94 (Bogoni et al., 2018). 101 Consequently, there is very scanty information on the effects of conservation of 102 habitats in the decrease of large- and medium-sized mammals within the southern 103 section of the biome. Data are retrieved from 65 camera traps installed during five 104 years, with a total effort of 29.788 camera-trap-days, in four protected areas and seven 105 forest fragments in Devonian knife ridges and surroundings areas in Southern Brazil, 106 aiming at quantifying defaunation in remnant forests of the southern region of the 107 Atlantic Rainforest to analyze the effects of fragmentation and forest conversion on the 108 structure of assemblages of large- and medium-sized mammals between the current 109 structure of assemblages compared with a historical baseline. We hypothesized that: (1) 110 in areas with a high percentage of natural environment conversion, mainly those 111 employed for agriculture and silviculture, defaunation rates are higher when compared 112 to areas with a greater percentage of natural forests; (2) all areas reveal a historical 113 erosion of local richness, averaging 70% of species (based on Bogoni et al. 2018). 114 115 MATERIAL AND METHODS 116 Study area 117 118 The current study sampled mammal fauna across 91 distribution places in 11 119 sampling sites of the Atlantic Rainforest of Southern Brazil (Fig. 1). Four are Protected 120 Areas (PAs), namely, Parque Nacional dos Campos Gerais (PCG), Reserva Biológica 121 das Araucárias (RBA), Reserva Particular do Patrimônio Natural Corredor do Iguaçu 122 (RPC) and Floresta Nacional de Irati (FNI). The other seven sites are non-protected 123 forest remnants, namely, Fragmento São Nicolau (FSN), Fragmento Portão Lajeado 124 (FPL), Fragmento Mirante (FMI), Fragmento Monte Seleto (FMS), Fragmento 125 Caxambu (FCA), Fragmento Barra Mansa (FBM) and Fragmento da Gruta (FGR) (Fig. 126 1, Table 1). Although sites RBA, RPC and FNI are not included within Environmental 127 Protection Area (EPA) limits of the Devonian Escarpment, they are taken into account 128 in current analysis due to their similar characteristics with regard to their phyto-129 physionomy and the use of soil of the other sampling sites sharing the surroundings and 130 comprising the meta-region of the Devonian Escarpment. 131 Sampling sites have vegetation composed of phyto-physionomies Mixed 132 Ombrophilous Forest (FOM) for sites RBA, FNI, RPC and FPL, and Height/Natural 133 Fields for PCG and FCA, coupled to transition zones between Height Fields and FOM 134 for sampling units FSN, FMI, FMS, FBM and FGR (Figure 1). According to Köppen´s 135 classification, the region´s climate is Cfa (humid temperate climate with hot summers) 136 for the sampling site RPC and Cfb (humid temperate climate with moderately hot 137 summers) for the other sampling units. Both are oceanic climates without a defined dry 138 season (Peel et al., 2007). 139 140 141 142 143 144 145 146 147 95 148 Figure 1. Map of the areas analyzed of the Atlantic Rainforest in southern Brazil, with 149 special reference to EPA of the Devonian Escarpment. PCG: Parque Nacional dos 150 Campos Gerais; RBA: Reversa Biológica das Araucárias; RPC: FNI: Floresta Nacional 151 de Irati; RPPN Corredor do Iguaçu; FSN: Fragmento São Nicolau; FPL: Fragmento 152 Portão lajeado; FMS: Fragmento Monte Seleto; FCA: Fragmento Caxambu; FMI: 153 Fragmento Mirante; FBM: Fragmento Barra Mansa; FGR: Fragmento da Gruta. 154 155 Mammal sampling and functional groups 156 157 Sampling sites were monitored at different times between 2011 and 2017. Sixty-158 five camera traps (Bushnell HD) were employed, with at least two devices in each 159 sampling unit, according to the size of each sampling area (Table 1). Camera traps were 160 distributed in the fragments, near water sprouts, on the treks and pathways, taking into 161 account the commonest trails used by species. Cameras were kept at an average distance 162 of 1,801 (± 792.7) and placed 50 cm above the ground on tree trunks and without baits. 163 Camera traps were active during at least 365 days of sampling (~8,760 h) at each site, 164 with a monthly change of memory cards and batteries during the study period. Sampling 165 effort was equal to the number of camera traps multiplied by the number of sampling 166 days (with 24 hours), resulting in a total effort of 29,788 trap-days (Table 1). An event 167 was independent when there were (a) consecutive photographs by the same camera with 168 an interval of at least 60 minutes or (b) nonconsecutive photographs by the same camera 169 (Srbek-Araujo and Chiarello, 2013) 170 The species Sus scroffa Linnaeus, 1758, Myocastor coypus (Molina, 1782) (see. 171 Pereira et al. 2020) and Lepus europaeus (Pallas, 1778), which are exotic and invasive 172 species in the region, were excluded. The species Sapajus nigritus (Goldfuss, 1809) and 173 Alouatta guariba clamitans Cabrera, 1940 (tree-dwelling species), rarely detected with 174 camera traps, were also excluded. 175 The alpha taxonomic nomenclature followed Paglia et al. (2012), following 176 96 recent taxonomic changes (e.g., Leopardus guttulus (Hensel, 1872), based on Trigo et 177 al. (2013)) and Sapajus genera, based on Lynch-Alfaro -Alfaro et al. (2012). In the case 178 of the orders Pilosa and Rodentia, Patton et al. (2015) were consulted for precise 179 identification. All those species with an adult body mass ≥1 kg were considered 180 medium- to large-bodied mammals (Paglia et al., 2012). 181 The species were classified into nine functional groups according to their 182 morpho-ecological traits which were not necessarily exclusive. Classification followed 183 suggestions on groups by Bogoni et al. (2018), or rather, species were classified 184 according to the following functional groups: (1) frugivores; (2) large grazers or 185 browsers: e.g., genus Mazama, Ozotoceros and Tapirus; (3) mesocarnivores (body mass 186 <13kg); (4) apex predator (>13kg). Following the previous classification, the proportion 187 of each main trophic category of each species was taken into account (sourced from 188 Wilman et al., 2004) to establish its energetic level (e.g. if a Tapirus population 189 consumes 70% leaves and 30% fruits, its trophic level will be 1.3 (i.e. (0.7 × 1) + (0.3 × 190 2)), grouping them according to the following functional groups: (5) high trophic level; 191 (6) intermediate trophic level and (7) low trophic level. Finally, the species were 192 grouped according to total biomass, included in the functional groups (8) small-bodied 193 species (<10kg) and (9) large-bodied species (≥10kg). Body mass thresholds and broad 194 trophic classes are based on Wilman et al. (2004) and Paglia et al. (2012). 195 196 Table 1. Description of sampling effort employed on all study areas in the Atlantic 197 Rainforest, Brazil. PCG: Parque Nacional dos Campos Gerais; RBA: Reversa Biológica 198 das Araucárias; RPC: FNI: Floresta Nacional de Irati; RPPN Corredor do Iguaçu; FSN: 199 Fragmento São Nicolau; FPL: Fragmento Portão lajeado; FMS: Fragmento Monte 200 Seleto; FCA: Fragmento Caxambu; FMI: Fragmento Mirante; FBM: Fragmento Barra 201 Mansa; FGR: Fragmento da Gruta. 202 203 Landscape characterization 204 205 Areas Total Traps Limit size (ha) Traps/ha (political size) (n) Linear distance (±SD) between traps (meters) Monitoring days Total Traps-Days Years sampling PCG 10 21.299 0.469 1.104±551.9 730 7.300 2013-2014 2016-2017 RBA 13 14.930 0.870 892±1.216 707 9.191 2012-2014 FNI 4 3.495 1.149 1.209±473.7 240 960 2012-2013 RPC 10 5.115 1.955 2.068±1.309 324 3.240 2011-2012 FSN 6 18.426 0.434 2.649±891 350 2.100 2015-2016 FPL 6 325 0.018 1438.5±1350.3 352 2.112 2015-2016 FMS 2 958 0.002 1.180±0 352 704 2015-2016 FCA 4 7.729 0.517 1.404±648.5 350 1.400 2015-2016 FMI 4 992 0.004 1.865±196.6 347 1.388 2015-2016 FBM 2 4.542 0.440 3.668±0 350 700 2015-2016 FGR 2 50 0.040 2.343±0 347 694 2015-2016 Total 65 - - - - 29.788 - Average 12 (±6.5) 0.536 (±0.57) 0.1563 (±0.38) 1,801 (±792.7) 111 (±107) 4099 (±1.577) - 97 Data retrieved from Brazilian ground charts, provided by the Instituto Brasileiro 206 de Geografia e Estatísticas (IBGE, 2018), were employed. Data include the 207 characterization of the landscape from OLI/Landsat-8 satellite images (spatial resolution 208 30 m) for the 2011-2016 period, on a 450x450 km grid (IBGE, 2018). 209 Each landscape was drawn as an area of 20 km2, including the extent of each 210 area, combining surrounding lands and covering 22,000 ha. The prevailing land cover 211 class was verified for each pixel (Hoskins et al., 2016). Cover classes and ground use 212 were defined as follows: Artificial areas (areas with predominant non-agricultural 213 anthropic surfaces); Silviculture (areas characterized by forest plantations of exotic or 214 native species as a monoculture); Agricultural areas (area characterized by temporal, 215 semi-perennial and permanent fields, irrigated or non-irrigated, with land used for the 216 production of food crops, fibers, and agribusiness commodities); Managed Pasture 217 (areas for the pasture of cattle and other animals, with cultivated vegetation (brachiaria, 218 ryegrass, and others) or grassland (natural), with intense anthropic interference; Forest 219 Vegetation (area with forests, except for monoculture of exotic vegetation) and 220 continental water bodies (including backland waters, such as rivers, streams, canals, and 221 other surface water bodies) (Table A1). Sampling sites were also categorized according 222 to the Human Development Index (HDI) and Demographic Density in the case of 223 municipalities in which they occur (Bogoni et al., 2018). 224 225 Data analysis 226 227 The defaunation index (Giacomini and Galetti, 2013) was used to assess the 228 degree of total mammal defaunation and for each forest remnant analyzed. The 229 defaunation index is a weighted measure of dissimilarity between focal assemblies and a 230 reference assembly (e.g., historical baseline). The index ranges between 0 (completely 231 intact) to 1 (completely defaunated) and is based on the Bray-Curtis dissimilarity index 232 with an importance weight species. In this case, all species have the same importance 233 rate (ω = 1) (see Giacomini and Galetti, 2013). The defaunation index was calculated 234 for the entire mammal assemblage (total defaunation) and each functional group 235 (assemblage-level) for each sampling unit through a matrix of presence and absence, 236 compared to the historical baseline. Baseline assemblage of current sampling sites was 237 employed to calculate the index, assuming probable occurrences based on known 238 geographic range polygons obtained from the IUCN (2019) to determine the historical 239 presence of each species at each sampling site (Bogoni et al., 2018). 240 We assessed whether landscape features (i.e., land use) and social variables (e.g., 241 IDH) affected observed total defaunation with Poisson generalized linear modeling 242 (GLM) and analysis of deviance based on the χ2 distribution (or quasi-Poisson GLM 243 and F-tests to account for over-dispersion; Zuur et al., 2009). The likelihood ratio test 244 was applied to select the model that best explained defaunation in relation to viable soil 245 use. The likelihood value is used to define the measure of fit, while the number of 246 parameters measures complexity (Zuur et al., 2009). Variables were removed one by 247 one, using the top-down strategy recommended by Diggle et al. (2002). Non-significant 248 variables (P>0.05) were tested with a more straightforward form (that is, with fewer 249 degrees of freedom) and, if the effect was non-significant, they were excluded from the 250 model (Zuur et al., 2009). 251 252 RESULTS 253 254 Thirty-nine mammal species were reported, retrieved from 7,755 independent 255 98 registers, ranging between 14 and 31 species per area, including discarded species 256 (Table A2). Baseline historical dataset revealed average (Dbs = 0.42) in defaunation 257 index for all sampling areas, with greatest defaunation for FGR, with (Dbs = 0.69), 258 followed by FPL (Dbs = 0.56) and FMI (Dbs = 0.55). PNC had the lowest defaunation 259 rates (Dbs = 0.14) (Fig. 2A). Protected areas had the lowest defaunation rates when 260 compared to non-protected ones (W = 861.5; P < 0.05) (Fig 2K.) 261 262 263 Fig 2. Defaunation index for medium- and large-bodied mammals on study sites in the 264 southern portion of the Brazilian Atlantic Forest biome: (a) all medium- and large-265 bodied mammal species; (b) high trophic level; (c) intermediate tropic level; (d) low 266 trophic level; (e) frugivores; (f) large grazers; (g) mesocarnivores; (h) apex-predators; 267 (i) small-bodied; (j) large-bodied; (k) boxplot illustrating the difference in defaunation 268 index between non-protected and protected areas. 269 270 271 In the case of functional groups, FGR had the highest defaunation rates for high 272 trophic level (Dbs = 0.60), intermediate trophic level (Dbs = 0.76), low trophic level (Dbs 273 = 0.70), mesocarnivores (Dbs = 0.46), apex-predators (Dbs = 1) and small-bodied (Dbs = 274 0.39). In the case of large grazers, FMI and FGR had the highest dafaunation rates (Dbs 275 = 0.85) in both areas. FMS revealed the highest defaunation rate for Frugivores (Dbs = 276 1), whilst defaunation for large-bodied mammals was highest at FMS (Dbs = 0.87) (Fig. 277 2, Table A3) 278 Within the context of the six land-use types and two socioeconomic variables 279 tested in the GLM, only silviculture, agriculture and vegetation better explained the total 280 defaunation rates in the sample units investigated (Table 2). GLM analysis explained 281 99 16% of total variation in mammal defaunation when only landscape variables were 282 taken into account. 283 284 Table 2. Results of GLM with a quasi-Poisson distribution of the best model for 285 defaunation according to the covered-land use variables. 286 287 Estimate SE t value P (Intercept) -0466795 0.189144 -2.468 <0.002 Silviculture 0.009585 0.001914 5.008 <0.001 Agriculture 0.007302 0.001913 3.818 <0.001 Forest Vegetation 0.012127 0.003260 3.720 <0.001 Dispersion parameter for quasi-Poisson Family taken to be 0.12464 Null deviance: 17.748 on 109 degrees of freedom 288 289 DISCUSSION 290 291 Our results showed a high defaunation degree within the Southern section of the 292 Brazilian Atlantic Rainforest, with more than 50% in all fragments analyzed. They 293 reveal defaunation means higher than the mean historical rate of 0.42 for the southern 294 section of the biome. Previous results indicated that the estimated defaunation rate for 295 the entire Atlantic Rainforest is even higher at 0.71 (See fig. 2 of Bogoni et al., 2018). 296 One of the main consequences of defaunation of large mammals at the local, regional 297 and global level is the loss of ecosystem services (Galetti and Dirzo, 2013; Dirzo et al., 298 2014; Galetti et al., 2015). Current results show that all sampled fragments are 299 undergoing consequences caused by defaunation in the composition of medium- and 300 large-sized functional mammal groups. Such a loss may result in unstable functional 301 fragments, with low potential resilience (Galetti and Dirzo, 2013; Kurten et al., 2015). 302 Within the functional groups analyzed, the Apex-predator species had the 303 greatest defaunation-caused decline. The species have essential ecosystem functions and 304 regulate and control the population of big herbivores, such as capybaras, deer and wild 305 pigs, and mesopredators such as opossum, tayra, coati, jaguaroundi (Jorge et al., 2013). 306 Their absence will probably interrupt the predator-prey relationships with unforeseen 307 effects in the ecosystemic functions (Weckel et al., 2006; Jorge et al., 2013). The big 308 jaguar is the biggest predator (50-160 kg) at the top of the neotropical region and its diet 309 is not superior to that of the puma (22-70 kg), the second biggest predator (Di Bitetti et 310 al., 2010). Although big jaguars have been historically distributed throughout the entire 311 Atlantic Rainforest biome (Beisiegel et al., 2012), it has been estimated that less than 312 300 adult specimens inhabit the biome, distributed in eight isolated populations 313 (Beisiegel et al., 2012; Paviolo et al., 2016). Despite the great sampling effort (29.788 314 camera-trap-days), with different search methodologies, the jaguar has not been 315 reported in any fragment under analysis. This indicates the local extinction of the 316 species. The leading causes for the population decrease of jaguars in the Brazilian 317 Southern Atlantic Rainforest, including the current region, is associated with habitat 318 loss, hunting and killing by ranchers who kill jaguars and pumas to retaliate loss in 319 stock herds (Paviolo et al., 2016; Srbek-Araujo and Chiarello, 2017). 320 The second most affected functional group by defaunation comprises the large 321 herbivores. Large grazers have a significant impact on ecosystemic processes and are 322 100 known to change the composition of vegetation (Elschot et al., 2015). Decrease or local 323 extinction of large grazers may trigger cascade events that alter the dynamics and 324 structure of local communities. Several studies have shown that grazing not only 325 reduces ground biomass but that grazer species function as ecosystem engineers. Due to 326 trampling, they increase the density in soil layers (Elschot et al., 2015, Nolte et al. 327 2013), causing anoxic conditions (i.e., lack of oxygen) and changes in the nitrogen cycle 328 of the soil (Schrama et al., 2013). 329 The third most affected functional group by defaunation lies at the intermediate 330 trophic level (Fig. 2). The group comprises peccaries, deer and tapirs, which have 331 unique ecological roles within the environment, such as the efficient removal of fruits, 332 long-distance dispersion and plant dispersion with large-sized seeds (Beck, 2006; Gayot 333 et al., 2004; O’Farrill et al., 2013). The southern section of the Atlantic Rainforest 334 experienced a severe collapse in mammal biomass, especially primary consumers 335 (Bogoni et al., 2018). Species such as Tayassu pecari and Tapirus terrestris (large 336 dispersers) showed low abundance in current inventory and demonstrated the rareness 337 of these species within the Environmental Protection Area (EPA) of the Devonian 338 Escarpment, indicating functional extinction or reduced activities. Both species are 339 classified as Critically Endangered in the state of Paraná, where the fragments under 340 analysis occur (MMA, 2014). They were intensely hunted in the past while they require 341 large areas for the maintenance of the population (Altrichter et al., 2012; Jorge et al., 342 2013). The local extinction of big frugivores has serious impacts on forest composition 343 and dynamics since seed dispersal is a key-step within the cycle of plant life, with 344 profound implications for succession, regeneration, carbon stock and conservation of 345 biodiversity (Bello et al., 2016; Peres et al., 2016). 346 After the analysis of variables related to soil use, the areas with the highest 347 percentage of soil cover for silviculture and agriculture proved to have the highest 348 defaunation levels when compared with areas with more forests and protected areas. As 349 conservation units, fragments had functionally balanced assemblies, with lower mean 350 rates of defaunation when compared to non-protected areas (Fig. 2K). Within the 351 remnants analyzed, the fragment Gruta (FGR), with the smallest area (23 ha) had the 352 highest defaunation rates. Although small fragments (<50 ha) may be viable stepping-353 stones between bigger fragments, and change landscape connectivity (Magioli et al., 354 2016), these fragments are incapable of sustaining viable mammal populations due to 355 size, quality and hunting pressure restrictions in the areas (Galetti et al., 2009; Canale et 356 al., 2012). 357 The Atlantic Rainforest is one of the most disturbed and fragmented ecosystems 358 in the Americas (Ribeiro et al., 2009). Its pristine area amounted to 1.5 million km², but 359 at present has a mere 11.7% of the original forest coverage, most of which are remnants 360 with less than 100 hectares surrounded by different types of agrosystem and silviculture 361 matrixes (Ribeiro et al., 2009). Effects of habitat fragmentation and natural forest 362 replacement by reforestation with exotic species on the community of large mammals 363 have already been investigated in studies at the local level (e.g., Bogoni et al., 2013; 364 2016; Wilson et al., 2016). Results indicate that matrix serves as a functional filter. 365 Most species are not able to obtain resources in such environments and remain confined 366 to smaller fragments (Chiarello, 2000). 367 Previous studies indicated that local extinctions are more pronounced at the 368 eastern sections of the Atlantic Rainforest (Northeastern and Eastern Atlantic 369 provinces), ranging between the states of Pernambuco and northern Minas Gerais, with 370 great repercussions on large mammals and species, appreciated by hunters past and 371 present (i.e., wild pig, jaguar, spider monkey, armadillo and anteater) (Canale et al., 372 101 2012; Mendes-Pontes et al., 2016; Bogoni et al., 2018). Current results show a similar 373 pattern for the southern section of the biome and indicate strong trends that the structure 374 for mammal assemblies is already compromised, due to large herbivore and carnivores 375 profoundly affected by defaunation. Further, even if hunting pressure and running down 376 of mammals in the region analyzed have not been evaluated, it may be inferred that in 377 this section of the Atlantic Rainforest, important changes in the functional composition 378 of the species are associated with landscape characteristics. 379 The conservation of biodiversity through the delimitation of new protected areas 380 in current political conditions in Brazil does not integrate state or federal management 381 and planning (Thomé and Haddad, 2019). New resources are thus highly necessary, 382 coupled to existing ones, for the promotion of activities foreseen in management of 383 existing conservation units that have not been fulfilled. These activities foresee the 384 management and control of exotic species within reserve areas and the units´ biological 385 restoration programs. If such measures are not taken, as large-sized mammals are 386 locally brought to extinction, pressures caused by anthropic changes will also affect 387 medium-sized animals and consequently influence abundance rates, with forest 388 remnants of the Atlantic Rainforest changing into empty and homogenous forests. 389 390 ACKNOWLEDGMENTS 391 392 The authors would like to thank the Postgraduate Program in Biological Sciences of the 393 Universidade Estadual de Londrina and the Coordination for the Upgrading of Higher 394 Education Personnel (CAPES-Funding Code 1689817), for logistical and financial 395 support. 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Springer, New York. 611 https://doi.org/10.1016/j.biocon.2009.02.021 https://doi.org/10.1007/s00442-012-2484-8 https://doi.org/10.1590/s1676-06032013000200005 https://doi.org/10.1017/S0030605315001222 https://doi.org/10.1111/j.1365-2028.2006.00705.x https://doi.org/10.1890/15-0885.1 https://doi.org/10.1126/science.aax9478 https://doi.org/10.1016/j.cub.2013.10.046 https://doi.org/10.1007/s10980-015-0312-3 106 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626 627 628 629 SUPPORT MATERIAL 630 107 Supporting Information S1 Landscape characterization of the analyzed areas. Areas: PCG: Parque Nacional dos Campos Gerais; RBA: Reversa Biológica das Araucárias; RPC: FNI: Floresta Nacional de Irati; RPPN Corredor do Iguaçu; FSN: Fragmento São Nicolau; FPL: Fragmento Portão lajeado; FMS: Fragmento Monte Seleto; FCA: Fragmento Caxambu; FMI: Fragmento Mirante; FBM: Fragmento Barra Mansa; FGR: Fragmento da Gruta; HB: Historical baseline. Areas Coordinates Silviculture Agriculture Artificial area Pasture Forest Vegetation Size area (ha) Demographic density Human Development Index Longitude Latitude PCG -50.0275 -25.0403 6.22 23.65 0 36.02 34,11 21.299 171.81 0.763 RBA -50.6146 -25.3005 9.16 55.95 9.16 0 32.86 14.930 42.43 0.66 FNI -50.5848 -25.3807 16.61 50.6 0.02 0 32.77 3.495 13.99 0.645 RPC -52.7147 -25.5158 48.1 16.75 1 30 4.15 5.115 19.39 0.629 FSN -50.0023 -24.1342 63.27 26.67 0 0 10.06 18.426 18.74 0.723 FPL -49.5609 -26.0214 29.53 68.71 0 0 1.76 325 25.94 0.686 FMS -49.6927 -25.9306 15.07 66.63 0.34 0 17.96 958 25.94 0.686 FCA -49.9792 -24.2835 50.58 41.29 0 7.2 0.93 7.729 18.74 0.708 FMI -49.3853 -24.3345 82 13.86 0.43 0 3.71 992 13.43 0.663 FBM -49.8177 -24.09 16.94 77.75 3.42 0.55 1.34 4.542 20.5 0.723 FGR -49.2693 -24.4006 82.45 0.09 0 0 17.46 50 13.43 0.663 108 Supporting Information S3 Land use cover found in forest remnants of the South Atlantic Forest in Brazil. 109 110 111 Supporting Information S2 List of medium-sized and large mammal species in the 11 areas analyzed along with the IUCN historical baseline. Trophic Guild: Ca: Carnivore; Fr: Frugivore; Fo: Folivore; Gr: Gumivore; Hb: Herbivore grazer; In: Insectivore; Myr: Myrmecophage; On: Omnivore; Os: Piscivore. Areas: PCG: Parque Nacional dos Campos Gerais; RBA: Reversa Biológica das Araucárias; RPC: FNI: Floresta Nacional de Irati; RPPN Corredor do Iguaçu; FSN: Fragmento São Nicolau; FPL: Fragmento Portão lajeado; FMS: Fragmento Monte Seleto; FCA: Fragmento Caxambu; FMI: Fragmento Mirante; FBM: Fragmento Barra Mansa; FGR: Fragmento da Gruta; HB: Historical baseline. 112 Taxon Considered Trophic Guild Areas HB Status IUCN PCG RBA FNI RPC FSN FPL FMS FCA FMI FBM FGR Didelphimorphia Didelphidae Didelphis albiventris Lund, 1840 Yes Fr/On X X X X X X X X X X LC Didelphis aurita (Wied-Neuwied, 1826) Yes Fr/On X X X X X X X X X X LC Pilosa Mymercophagidae Myrmecophaga tridactyla Linnaeus, 1758 Yes Myr X X X X X X X VU Tamandua tetradactyla (Linnaeus, 1758) Yes Myr X X X X X X X X X LC Cingulata Dasypodidae Cabassous tatouay (Desmarest, 1804) Yes Myr X X X X X X LC Dasypus novemcinctus Linnaeus, 1758 Yes In/On X X X X X X X X X X X X LC Dasypus septemcinctus Linnaeus, 1758 Yes In/On X X X LC Euphractus sexcinctus (Linnaeus, 1758) Yes In/On X X X X X LC Perissodactyla Tapiridae Tapirus terrestris (Linnaeus, 1758) Yes Hb/Fr X X VU Cetartiodactyla Cervidae Mazama americana (Erxleben, 1777) Yes Fr/Hb X X X X X X X X X X DD Mazama gouazoubira (G. Fischer, 1814) Yes Fr/Hb X X X X X X X X X X X X LC Mazama nana (Hensel,1872) Yes Fr/Hb X X VU Ozotoceros bezoarticus (Linnaeus, 1758) Yes Fr/Hb X NT Tayassuidae Pecari tajacu (Linnaeus, 1758) Yes Fr/Hb X X X X X X X X LC Tayassu pecari (Link, 1795) Yes Fr/Hb X X X VU 113 Suidae Sus scrofa Linnaeus, 1758 * No On X X Primates Atelidae Alouatta guariba clamitans Cabrera, 1940 No Fo/Fr X X X X X X X X X Cebidae Sapajus nigritus (Goldfuss, 1809) No Fr/On X X X X X X X X X Carnivora Canidae Cerdocyon thous (Linnaeus, 1766) Yes In/On X X X X X X X X X X X X LC Chrysocyon brachyurus (Illiger, 1815) Yes Ca/On X X X X X X NT Speothos venaticus (Lund, 1842) Yes Ca X NT Felidae Leopardus guttulus (Hensel, 1872) Yes Ca X X X X X X X X X X X X VU Leopardus pardalis (Linnaeus, 1758) Yes Ca X X X X X X X X X X X X LC Leopardus wiedii (Schinz, 1821) Yes Ca X X X X X X X X X X NT Panthera onca (Linnaeus, 1758) Yes Ca X NT Puma concolor (Linnaeus, 1771) Yes Ca X X X X X X X X X X X LC Herpailurus yagouaroundi (É. Geoffroy Saint-Hilaire, 1803) Yes Ca X X X X X X X X X LC Mustelidae Eira barbara (Linnaeus, 1758) Yes Fr/On X X X X X X X X X X X LC Galictis cuja (Molina, 1782) Yes Ca X X X X X X X LC Lontra longicaudis (Olfers, 1818) No Ps X X X NT Procyonidae Nasua nasua (Linnaeus, 1766) Yes Fr/On X X X X X X X X X X X X VU Procyon cancrivorus (G. Cuvier,1798) Yes Fr/On X X X X X X X X X X X LC Lagomorpha Leporidae 114 Sylvilagus brasiliensis (Linnaeus, 1758) Yes Hb X X X X X EN Lepus europaeus Pallas, 1778* No Hb X X X X X X X X X Rodentia Caviidae Hydrochoerus hydrochaeris (Linnaeus, 1766) No Hb X X X X X X X X LC Cuniculidae Cuniculus paca (Linnaeus, 1766) Yes Fr/Hb X X X X X X X X LC Dasyproctidae Dasyprocta azarae Lichtenstein, 1823 Yes Fr/Gr X X X X X X X X X X X X DD Echimyidae Myocastor coypus (Molina, 1782) No X X Erethizontidae Fr/On Coendou spinosus (F. Cuvier, 1823) Yes Fr/Fo X X X X X X X LC 115 Supporting Information S3 Defaunation values found in forest remnants of the South Atlantic Forest in Brazil. Areas DEFAUNATION Historical High trophic level Intermediat e trophic level Low trophic level Frugivores Large grazers Mesocarnivores Apex predator Small- bodied species Large- bodied species PCG 0.14 0.30 0.05 0.17 0.07 0.06 0.21 0.53 0.00 0.22 RBA 0.46 0.48 0.43 0.55 0.07 0.65 0.21 0.53 0.11 0.53 FNI 0.43 0.44 0.49 0.25 0.59 0.48 0.21 0.53 0.18 0.53 RPC 0.18 0.44 0.11 0.02 0.00 0.10 0.21 0.53 0.14 0.20 FSN 0.30 0.32 0.42 0.02 0.00 0.48 0.21 0.53 0.07 0.00 FPL 0.56 0.55 0.53 0.66 0.59 0.65 0.21 0.53 0.21 0.87 FMS 0.56 0.46 0.56 0.34 1.00 0.48 0.35 0.53 0.21 1.00 FCA 0.40 0.34 0.52 0.19 0.07 0.48 0.30 0.53 0.33 0.22 FMI 0.55 0.35 0.71 0.55 0.07 0.85 0.21 0.53 0.29 0.42 FBM 0.38 0.38 0.45 0.19 0.00 0.48 0.30 0.53 0.23 0.22 FGR 0.69 0.60 0.76 0.70 0.59 0.85 0.46 1.00 0.39 0.42 116 CONCLUSÃO GERAL O presente estudo foi o primeiro a avaliar o grau de defaunação em remanescentes de Mata Atlântica Sul in loco, localizados no estado do Paraná. Mais especificamente o foco de nosso interesse foi a região da Área de Proteção Ambiental da Escarpa Devoniana. Esta importante área de conservação da biodiversidade brasileira, vem sofrendo intensas pressões nas esferas legislativas e do setor agropecuário para a redução de sua área de extensão geográfica. Alguns desses setores alegam que a atual demarcação já contempla áreas com atividades agrícolas entre outras e a sua redução não implicaria em danos à conservação das espécies dos remanescentes. Nós concluímos que os remanescentes de Mata Atlântica sul enfrentam diretamente questões indicadas como precursoras da defaunação em todo o globo. A primeira é o desconhecimento básico sobre distribuição geográfica das espécies em remanescentes florestais. Este problema é ainda mais grave ao consideramos que existem unidades de conservação que não contam sequer com inventários básicos da diversidade, dificultando assim qualquer plano de conservação, manejo ou estratégias de restauração. A segunda questão é a mortalidade de animais em pequenas rodovias que cortam remanescentes florestais. Nossos dados sugerem que este é um fator que deve ser considerado como causa direta para o declínio de algumas espécies, sendo necessário a implementação de medidas de mitigação básicas que constam na legislação do Brasil e do Estado do Paraná. A terceira questão é a introdução de espécies não nativas tendo como consequência eventos de invasões biológicas. Já está bem relatado os impactos ambientais e econômicos do javali (Sus scrofa) no território brasileiro. Porém pouco se sabe do impacto aos ecossistemas que o ratão-do-banhado (Mycastor coypus) pode causar nos diferentes sistemas biológicos do Brasil. Nossos dados indicam que a espécie pode aumentar seu limite de ocorrência vindo a se tornar uma “espécie problema” se medidas de controle não forem adotadas em curto espaço de tempo. Por fim nossos dados indicam que o longo histórico de conversão de florestas naturais em pastagens e monoculturas de espécies não nativas, reflete diretamente o grau de defaunação no remanentes restantes. Mesmo em áreas densamente fragmentadas os mamíferos de médio e grande porte persistem na região da APA da Escarpa Devoniana bem como em regiões do segundo planalto meridional paranaense. Contudo é possível observar em todos os remanescentes florestais analisados eventos de defaunação local. Podemos denotar que em áreas protegidas a defaunação é menor se comparadas a remanescentes sem qualquer status de 117 proteção. Ainda assim, salientamos que desenho experimental pode ser melhorado, através da inclusão de mais remanecentes de Mata Atlântica em diferentes estados de conservação e com matrizes diversifiucadas. O declínio e ausência de espécies chaves em determinadas regiões com interesse conservacionista, indicam que as florestas estão sob risco de instabilidade funcional, acarretando na perda direta de serviços necessários para a sua manutenção e resiliência. 118 ANEXO A Normas técnicas – Revista Papeis Avulsos de Zoologia 22/02/2020 Submissions | Papéis Avulsos de Zoologia www.revistas.usp.br/paz/about/submissions#authorGuidelines 1/8 PORTAL DE REVISTAS DA USP Search CURRENT ARCHIVES ANNOUNCEMENTS ABOUT Register Login HOME Submissions Login or Register to make a submission. 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All articles that contain nomenclatural acts http://www.scielo.br/pdf/paz/term%20of%20assent%20and%20cession%20of%20copyright%20(pa).docx https://creativecommons.org/licenses/by/4.0 https://www.councilscienceeditors.org/resource-library/editorial-policies/white-paper-on-publication-ethics/2-1-editor-roles-and-responsibilities/#214 https://www.abecbrasil.org.br/arquivos/whitepaper_CSE.pdf https://www.force11.org/group/fairgroup/fairprinciples http://www.ncbi.nlm.nih.gov/genbank https://www.embl.org/ 22/02/2020 Submissions | Papéis Avulsos de Zoologia www.revistas.usp.br/paz/about/submissions#authorGuidelines 6/8 are registered in Zoobank by the journal staff and the Life Science Identifier (LSID) of the article is included in the published version. Authors are encouraged to deposit their data in a repository that is best suited to their dataset. Some examples of repositories are the Global Biodiversity Information Facility (GBIF), Environmental Data Initiative (EDI), Dryad, Figshare, Zenodo, etc. The data repository used must guarantee the preservation of the data and provide a persistent identifier such as a DOI so it can be accessible and citable. Journal will make exceptions for sensitive information as the location of endangered species. Datasets used in the research, deposited in a scientific research data repository, should be cited in the “Materials and method section” and its reference (with the DOI number) should be included in the reference list. Datasets References Examples: Dataset deposited in a scientific research data repository: Botham, M.; Roy, D.; Brereton, T.; Middlebrook, I.; Randle, Z. 2013. United Kingdom Butterfly Monitoring Scheme: species trends 2012. NERC Environmental Information Data Centre. [dataset]. Available at: https://doi.org/10.5285/5afbbd36-2c63-4aa1-8177-695bed98d7a9 . Accessed: 13/04/2019. United States Department of Health and Human Services. Substance Abuse and Mental Health Services Administration. Office of Applied Studies. 2015. Treatment Episode Data Set - - Discharges (TEDS-D) -- Concatenated, 2006 to 2011. Version V5. Ann Arbor, MI: Inter- university Consortium for Political and Social Research [distributor], 23 Nov. 2015. Available at: https://doi.org/10.3886/ICPSR30122.v5 . Accessed: 30/09/2019.  Article supplementary information deposited in a scientific research data repository: Yoon, J; Sofaer, H.R, Sillet, T. S, Morrison, S.A., Ghalambor, C.K. 2017. Data from: The relationship between female brooding and male nestling provisioning: does climate underlie geographic variation in sex roles?. Journal of Avian Biology, 48(2):220-228. Available at: https://doi.org/10.5285/5afbbd36-2c63-4aa1-8177-695bed98d7a9 . Accessed: 07/07/2019. Dataset published as data paper: Bovendorp, R. S., Villar, N., de Abreu-Junior, E. F., Bello, C., Regolin, A. L., Percequillo, A. R., & Galetti, M. 2017. Atlantic small-mammal: a dataset of communities of rodents and marsupials of the Atlantic forests of South America. Ecology, 98(8):2226–2226. [data paper]. Available at: http://dx.doi.org/10.1002/ecy.1893. Accessed: 07/10/2019. AUTHOR SELF-ARCHIVING Authors can share the accepted manuscript¹ or the published version² of the manuscript with their colleagues and post them on personal websites or institutional repositories for academic purposes while providing bibliographic details that credit, if applicable, its publication in this journal. ¹ Accepted manuscript - sometimes called post-print version, is the final draft author manuscript including the referees’ suggestions, before the copyediting process. ² Published version - is the article published in PDF format available at the Papéis Avulsos de Zoologia website. http://www.zoobank.org/ https://www.gbif.org/ https://portal.edirepository.org/nis/home.jsp https://datadryad.org/stash https://figshare.com/ https://zenodo.org/ https://doi.org/10.5285/5afbbd36-2c63-4aa1-8177-695bed98d7a9 https://doi.org/10.3886/ICPSR30122.v5 https://doi.org/10.5285/5afbbd36-2c63-4aa1-8177-695bed98d7a9 http://dx.doi.org/10.1002/ecy.1893 22/02/2020 Submissions | Papéis Avulsos de Zoologia www.revistas.usp.br/paz/about/submissions#authorGuidelines 7/8 PUBLICATION ETHICS Research or publication misconduct (plagiarism, self-plagiarism, fabrication or falsification of data and results, etc.) will be treated in accordance with the Commission on Publication Ethics (COPE) guidelines. Allegations, corrections, and doubts must be sent to the Editor (contact information). All allegations/cases will be evaluated and, when necessary, retractions, corrections or expressions of concern will be published. CONTENT For more details of the manuscript preparation format, see CBE Style Manual, available at the electronic address of the Council of Science Editors. Papéis Avulsos de Zoologia is a publication of the Museu de Zoologia da Universidade de São Paulo. Always consult the instructions to authors updated on the electronic pages: www.scielo.br/paz or www.revistas.usp.br/paz. Copyright Notice Responsibility: The scientific content and the opinions expressed in the manuscript are the sole responsibility of the author(s). Copyrights: Papéis Avulsos de Zoologia. The journal is licensed under CC-BY Creative Commons license. Privacy Statement The names and email addresses entered in this journal site will be used exclusively for the stated purposes of this journal and will not be made available for any other purpose or to any other party. MAKE A SUBMISSION KEYWORDS https://publicationethics.org/guidance/Guidelines https://www.revistas.usp.br/paz/about/contact http://www.councilscienceeditors.org/publications/scientific-style-and-format http://www.mz.usp.br/ http://www.scielo.br/paz http://www.revistas.usp.br/paz http://creativecommons.org/ http://www.revistas.usp.br/paz/about/submissions 126 ANEXO B Normas técnicas – Revista Biological Conservation AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 1 BIOLOGICAL CONSERVATION AUTHOR INFORMATION PACK TABLE OF CONTENTS . XXX . • Description • Audience • Impact Factor • Abstracting and Indexing • Editorial Board • Guide for Authors p.1 p.1 p.1 p.2 p.2 p.4 ISSN: 0006-3207 DESCRIPTION . Biological Conservation is a leading international journal in the discipline of conservation science. The journal publishes articles spanning a diverse range of fields that contribute to the biological, sociological, ethical and economic dimensions of conservation. The primary aim of Biological Conservation is the publication of high-quality papers that advance the science and practice of conservation, or which demonstrate the application of conservation principles and policy. Biological Conservation invites the submission of research articles, reviews (including systematic reviews and perspectives), short communications, policy perspectives, and letters to the editor dealing with all aspects of conservation science, including theoretical and empirical investigations into the consequences of human actions for the diversity, structure and function of terrestrial, aquatic or marine ecosystems. Such papers may include quantitative assessments of extinction risk, fragmentation effects, spread of invasive organisms, conservation genetics, conservation management, global change effects on biodiversity, landscape or reserve design and management, restoration ecology, or resource economics. We also welcome papers coming from social sciences including those reporting on advances in conservation politics, ethics, policy, human social structure and biodiversity, and political culture among other subjects. Biological Conservation covers interdisciplinary topics within conservation biology and also provides practical applications of conservation research for land/resource managers and policy makers. We publish articles and thematic special issues that have a global relevance in terms of the topics or issues addressed, and thus demonstrate applications of conservation science and management beyond the specific system or species studied.Biological Conservation is an affiliate publication of the Society for Conservation Biology (SCB). SCB members can obtain a personal subscription to this journal through the Society. AUDIENCE . Environmentalists, conservationists, botanists, marine scientists, ecologists, biologists, zoologists. IMPACT FACTOR . 2018: 4.451 © Clarivate Analytics Journal Citation Reports 2019 https://ees.elsevier.com/bioc/ https://www.journals.elsevier.com/biological-conservation/editorial-board/ http://www.conbio.org/publications/affiliate-publications AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 2 ABSTRACTING AND INDEXING . Environmental Periodicals Bibliography Current Advances in Ecological Sciences AGRICOLA Embase Engineering Village - GEOBASE Current Contents - Agriculture, Biology & Environmental Sciences Energy Information Abstracts Biological and Agricultural Index Science Citation Index Cambridge Scientific Abstracts Elsevier BIOBASE Scopus EDITORIAL BOARD . Editor-in-Chief Vincent Devictor, UMR CNRS-UM2 5554 cc065, Institut des Sciences de l'Evolution de Montpellier Editors Amanda Bates, Memorial University of Newfoundland, St John's, Newfoundland and Labrador, Canada Richard Corlett, Chinese Academy of Sciences, Menglun, Yunnan, China Graeme Cumming, ARC Centre of Excellence for Coral Reef Studies, Townsville, Queensland, Australia Varun R. Goswami, Conservation Initiatives, Guwahati, India Lian Pin Koh, Betty and Gordon Moore Center for Science Conservation International, Arlington, Virginia, United States Rafael Loyola, Federal University of Goias, Brazil Bea Maas, University of Vienna, Wien, Austria Anna Pidgeon, University of Wisconsin-Madison Department of Forest and Wildlife Ecology, Madison, Wisconsin, United States Richard B. Primack, Boston University, Boston, Massachusetts, United States Tracey Regan, Arthur Rylah Institute for Environmental Research, Heidelberg, Victoria, Australia Robin Roth, University of Guelph, Guelph, Ontario, Canada Book Review Editor David Johns, PO Box 725, McMinneville, Oregon, OR 97218 Editorial Board Barry Brook, The University of Adelaide, Adelaide, South Australia, Australia Zuzana Burivalova, Princeton University, Princeton, New Jersey, United States Regis Cereghino, Toulouse 1 University Capitole, Toulouse, France Jin Chen, Xishuangbanna Tropical Botanical Garden, Mengla, China Enrico Di Minin, University of Helsinki, Helsinki, Finland Johan Ekroos, Lund University, Lund, Sweden Markus Fischer, University of Bern, Bern, Switzerland Travis Gallo, George Mason University, Fairfax, Virginia, United States Kevin Gaston, University of Exeter Environment and Sustainability Institute, Penryn, United Kingdom Laurent Godet, Coast Environment Remote Sensing Geomatics Nantes, Nantes, France Radim Hédl, Institute of Botany Czech Academy of Sciences Department of Vegetation Ecology, Brno, Czech Republic Zhigang Jiang, Chinese Academy of Sciences, Beijing, China Janice Ser Huay Lee, Nanyang Technological University, Singapore, Singapore Pia Lentini, University of Melbourne School of BioSciences, University Of Melbourne, Australia David Lindenmayer, Australian National University, Canberra, Australia Hong Liu, Florida International University, Miami, Florida, United States Tessa Mazor, University of Queensland, Brisbane, Queensland, Australia Abraham Miller-Rushing, Acadia National Park, Bar Harbor, Maine, United States Peter Moyle, University of California Davis, Davis, California, United States Javier Nori, National University of Cordoba, Cordoba, Argentina Daniel Oro, Mediterranean Institute for Advanced Studies, Esporles, Spain Ludmila Rattis, Woods Hole Research Center, Falmouth, Massachusetts, United States Denis Allan Saunders, CSIRO Land and Water Floreat, Floreat, Australia Christian H. Schulze, University of Vienna Faculty of Life Sciences, Vienna, Austria AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 3 Anna Sher, University of Denver, Denver, Colorado, United States Assaf Shwartz, Technion Israel Institute of Technology, Haifa, Israel Steve Sinclair, Arthur Rylah Institute for Environmental Research, Heidelberg, Victoria, Australia Fernanda Thiesen Brum, Federal University of Parana - Polytechnic Centre Campus, Curitiba,Paraná, Brazil Anne Toomey, Pace University, New York, New York, United States Serge Wich, Liverpool John Moores University, Liverpool, United Kingdom AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 4 GUIDE FOR AUTHORS . Your Paper Your Way We now differentiate between the requirements for new and revised submissions. You may choose to submit your manuscript as a single Word or PDF file to be used in the refereeing process. Only when your paper is at the revision stage, will you be requested to put your paper in to a 'correct format' for acceptance and provide the items required for the publication of your article. To find out more, please visit the Preparation section below. INTRODUCTION Please read all information carefully and follow the instructions in detail when preparing your manuscript. Manuscripts that are not prepared according to our guidelines will be sent back to authors without review. Biological Conservation encourages the submission of high-quality manuscripts that advance the science and practice of conservation, or which demonstrate the application of conservation principles and policy. Conservation implications should be clearly emphasized and discussed. Given the broad international readership of the journal, published articles should have global relevance in terms of the topics or issues addressed, and thus demonstrate applications for conservation beyond the specific system or species studied. Types of paper Word counts include text, references, figures and tables. Each figure or table should be considered equal to 300 words. 1. Full length articles (Research papers) Research papers report the results of original research. The material must not have been previously published elsewhere. Full length articles are up to 8,000 words. 2. Review articles Reviews summarize the status of research in a field of current interest. They may be submitted or invited. Review articles are up to 12,000 words and must include a methods section explaining how the literature for review was selected. We also consider Systematic Reviews, which apply a methodology to synthesize and appraise the scientific evidence on a specific question or hypothesis. More about systematic reviews can be found here: http://www.environmentalevidence.org/information-for-authors. 3. Perspectives: These articles provide an opportunity for authors to present a novel, distinctive viewpoint on any subject within the journal's scope. The article should be well grounded in evidence and adequately supported by citations but may focus on a stimulating and thought-provoking line of argument that represents a significant advance in thinking about conservation problems and solutions. Perspectives articles should not exceed 8000 words. 4. Short communications Short communications highlight both novel research and replication studies that report preliminary findings that are particularly compelling and highly relevant to conservation science and practice. If submitting a replication study, please include in your cover letter the rationale for undertaking the study. Short communications should not exceed 4,000 words. 5. Policy Analysis These are short commentary pieces on contemporary, internationally relevant conservation or conservation-related policy issues that enable researchers, policy makers, and practitioners to make timely contributions to policy debates and actions. Contributions are based on research, expert analysis, literature review, or practitioner reflections regarding specific policy issues. Pure opinion pieces will not be considered for this paper type. Forum articles should be written in an accessible style and supported by real world examples and/or referenced scientific evidence and should not exceed 4,000 words. AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 5 6. Fast-Tracked Papers Fast-Tracked Papers may be any of the above paper types, but are to be submitted only upon invitation from the editors. These papers will be fast-tracked by having reviewers lined up by the handling editor in advance, and upon publication these papers will be showcased in an ongoing special issue. 7. Book Reviews Book reviews will be included in the journal on a range of relevant titles that are not more than two years old. These are usually less than 2,000 words. Please submit your requests/ideas to David Johns at johnsd@embarqmail.com. 8. Editorials Opinion pieces by experts on a topic, usually invited by the Editor. The topic is usually timely and offers important insights into the field. 9. Correspondence Letters to the Editor (Correspondence) papers are responses to recently published papers. Letters must be short (a maximum 800 words) and include only key references (5 maximum) and one figure if necessary. The content must be constructive, discuss significant issues, and respectful in tone. Provided the editors agree that publication of the letter is warranted, it will generally also receive a response from the authors of the original article, and both letter and response will be published in the same issue. Submission checklist You can use this list to carry out a final check of your submission before you send it to the journal for review. Please check the relevant section in this Guide for Authors for more details. Ensure that the following items are present: One author has been designated as the corresponding author with contact details: • E-mail address • Full postal address All necessary files have been uploaded: Manuscript: • Include keywords • All figures (include relevant captions) • All tables (including titles, description, footnotes) • Ensure all figure and table citations in the text match the files provided • Indicate clearly if color should be used for any figures in print Graphical Abstracts / Highlights files (where applicable) Supplemental files (where applicable) Further considerations • Manuscript has been 'spell checked' and 'grammar checked' • All references mentioned in the Reference List are cited in the text, and vice versa • Permission has been obtained for use of copyrighted material from other sources (including the Internet) • A competing interests statement is provided, even if the authors have no competing interests to declare • Journal policies detailed in this guide have been reviewed • Referee suggestions and contact details provided, based on journal requirements • We strongly encourage authors to check the existing evidences for their case at Conservation Evidence. The introduction of the paper should include a sentence highlighting whether any evidence is already available or not with corresponding references. For further information, visit our Support Center. BEFORE YOU BEGIN Ethics in publishing Please see our information pages on Ethics in publishing and Ethical guidelines for journal publication. https://www.conservationevidence.com/ https://www.conservationevidence.com/ http://service.elsevier.com/app/home/supporthub/publishing/ https://www.elsevier.com/about/policies/publishing-ethics https://www.elsevier.com/authors/journal-authors/policies-and-ethics AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 6 Policy and Ethics All appropriate ethics and other approvals were obtained for the research. Where appropriate, authors should state that their research protocols have been approved by an authorized animal care or ethics committee, and include a reference to the code of practice adopted for the reported experimentation or methodology. The Editor will take account of animal welfare issues and reserves the right not to publish, especially if the research involves protocols that are inconsistent with commonly accepted norms of animal research. Declaration of interest All authors must disclose any financial and personal relationships with other people or organizations that could inappropriately influence (bias) their work. Examples of potential conflicts of interest include employment, consultancies, stock ownership, honoraria, paid expert testimony, patent applications/ registrations, and grants or other funding. Authors should complete the declaration of interest statement using this template and upload to the submission system at the Attach/Upload Files step. If there are no interests to declare, please choose: 'Declarations of interest: none' in the template. This statement will be published within the article if accepted. More information. Submission declaration and verification Submission of an article implies that the work described has not been published previously (except in the form of an abstract, a published lecture or academic thesis, see 'Multiple, redundant or concurrent publication' for more information), that it is not under consideration for publication elsewhere, that its publication is approved by all authors and tacitly or explicitly by the responsible authorities where the work was carried out, and that, if accepted, it will not be published elsewhere in the same form, in English or in any other language, including electronically without the written consent of the copyright- holder. To verify originality, your article may be checked by the originality detection service Crossref Similarity Check. Use of inclusive language Inclusive language acknowledges diversity, conveys respect to all people, is sensitive to differences, and promotes equal opportunities. Articles should make no assumptions about the beliefs or commitments of any reader, should contain nothing which might imply that one individual is superior to another on the grounds of race, sex, culture or any other characteristic, and should use inclusive language throughout. Authors should ensure that writing is free from bias, for instance by using 'he or she', 'his/her' instead of 'he' or 'his', and by making use of job titles that are free of stereotyping (e.g. 'chairperson' instead of 'chairman' and 'flight attendant' instead of 'stewardess'). Author contributions For transparency, we encourage authors to submit an author statement file outlining their individual contributions to the paper using the relevant CRediT roles: Conceptualization; Data curation; Formal analysis; Funding acquisition; Investigation; Methodology; Project administration; Resources; Software; Supervision; Validation; Visualization; Roles/Writing - original draft; Writing - review & editing. Authorship statements should be formatted with the names of authors first and CRediT role(s) following. More details and an example Changes to authorship Authors are expected to consider carefully the list and order of authors before submitting their manuscript and provide the definitive list of authors at the time of the original submission. Any addition, deletion or rearrangement of author names in the authorship list should be made only before the manuscript has been accepted and only if approved by the journal Editor. To request such a change, the Editor must receive the following from the corresponding author: (a) the reason for the change in author list and (b) written confirmation (e-mail, letter) from all authors that they agree with the addition, removal or rearrangement. In the case of addition or removal of authors, this includes confirmation from the author being added or removed. Only in exceptional circumstances will the Editor consider the addition, deletion or rearrangement of authors after the manuscript has been accepted. While the Editor considers the request, publication of the manuscript will be suspended. If the manuscript has already been published in an online issue, any requests approved by the Editor will result in a corrigendum. https://www.elsevier.com/declaration-of-competing-interests http://service.elsevier.com/app/answers/detail/a_id/286/supporthub/publishing https://www.elsevier.com/authors/journal-authors/policies-and-ethics https://www.elsevier.com/authors/journal-authors/policies-and-ethics https://www.elsevier.com/editors/perk/plagiarism-complaints/plagiarism-detection https://www.elsevier.com/editors/perk/plagiarism-complaints/plagiarism-detection https://www.elsevier.com/authors/journal-authors/policies-and-ethics/credit-author-statement AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 7 Article transfer service This journal is part of our Article Transfer Service. This means that if the Editor feels your article is more suitable in one of our other participating journals, then you may be asked to consider transferring the article to one of those. If you agree, your article will be transferred automatically on your behalf with no need to reformat. Please note that your article will be reviewed again by the new journal. More information. Copyright Upon acceptance of an article, authors will be asked to complete a 'Journal Publishing Agreement' (see more information on this). An e-mail will be sent to the corresponding author confirming receipt of the manuscript together with a 'Journal Publishing Agreement' form or a link to the online version of this agreement. Subscribers may reproduce tables of contents or prepare lists of articles including abstracts for internal circulation within their institutions. Permission of the Publisher is required for resale or distribution outside the institution and for all other derivative works, including compilations and translations. If excerpts from other copyrighted works are included, the author(s) must obtain written permission from the copyright owners and credit the source(s) in the article. Elsevier has preprinted forms for use by authors in these cases. For gold open access articles: Upon acceptance of an article, authors will be asked to complete an 'Exclusive License Agreement' (more information). Permitted third party reuse of gold open access articles is determined by the author's choice of user license. Author rights As an author you (or your employer or institution) have certain rights to reuse your work. More information. Elsevier supports responsible sharing Find out how you can share your research published in Elsevier journals. Role of the funding source You are requested to identify who provided financial support for the conduct of the research and/or preparation of the article and to briefly describe the role of the sponsor(s), if any, in study design; in the collection, analysis and interpretation of data; in the writing of the report; and in the decision to submit the article for publication. If the funding source(s) had no such involvement then this should be stated. Open access Please visit our Open Access page from the Journal Homepage for more information. Elsevier Researcher Academy Researcher Academy is a free e-learning platform designed to support early and mid-career researchers throughout their research journey. The "Learn" environment at Researcher Academy offers several interactive modules, webinars, downloadable guides and resources to guide you through the process of writing for research and going through peer review. Feel free to use these free resources to improve your submission and navigate the publication process with ease. Language (usage and editing services) Please write your text in good English (American or British usage is accepted, but not a mixture of these). Authors who feel their English language manuscript may require editing to eliminate possible grammatical or spelling errors and to conform to correct scientific English may wish to use the English Language Editing service available from Elsevier's Author Services. Submission Our online submission system guides you stepwise through the process of entering your article details and uploading your files. The system converts your article files to a single PDF file used in the peer-review process. Editable files (e.g., Word, LaTeX) are required to typeset your article for final publication. All correspondence, including notification of the Editor's decision and requests for revision, is sent by e-mail. https://www.elsevier.com/authors/article-transfer-service https://www.elsevier.com/about/policies/copyright https://www.elsevier.com/about/policies/copyright/permissions https://www.elsevier.com/__data/assets/word_doc/0007/98656/Permission-Request-Form.docx https://www.elsevier.com/about/policies/copyright https://www.elsevier.com/about/policies/open-access-licenses https://www.elsevier.com/about/policies/copyright https://www.elsevier.com/about/policies/copyright https://www.elsevier.com/authors/journal-authors/submit-your-paper/sharing-and-promoting-your-article https://researcheracademy.elsevier.com/ http://webshop.elsevier.com/languageediting/ http://webshop.elsevier.com/languageediting/ AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 8 Referees Authors are at liberty to suggest the names of up to three potential reviewers (with full contact details). Potential reviewers should not include anyone with whom the authors have collaborated during the research being submitted. Additional Information Editorial Process Publishing space in the journal is limited, such that many manuscripts must be rejected. To expedite the processing of manuscripts, the journal has adopted a two-tier review process. During the first stage of review, the handling editor evaluates the manuscript for appropriateness and scientific content, taking advice where appropriate from members of the editorial board. Criteria for rejection at this stage include: Manuscript lacks a strong conservation focus or theme, or management implications not well-developed. Please note that research on a rare or endangered species or ecosystem is not sufficient justification to merit publication in Biological Conservation. Published research must also advance the science and practice of conservation biology, and thus have broader application for a wide international audience.Manuscript subject matter more appropriate for another journal. Natural history or biodiversity surveys, including site descriptions, are usually better suited for other outlets, such as a regional or taxon-specific journal. Similarly, manuscripts with a primarily behavioral, genetic or ecological focus are more appropriate for journals in those fields. For example, studies reporting on disturbance effects, species interactions (e.g., predator- prey, competitive, or pollinator-host plant interactions), species-habitat relationships, descriptive genetics (e.g., assays of genetic variation within or between populations), or behavioral responses to disturbance will be referred elsewhere if they lack a clear conservation message. Authors are advised to contact an Editor prior to submission if there are any questions regarding the appropriateness of a manuscript for the journal.Study primarily of local or regional interest. Biological Conservation is international in scope, and thus research published in the journal should have global relevance, in terms of the topics or issues addressed.Study poorly designed or executed. Research lacks spatial or temporal replication, has insufficient sample sizes, or inadequate data analysis. Such obvious indications of poor-quality science will be cause for immediate rejection.Manuscript poorly written. Poor writing interferes with the effective communication of science. Authors for whom English is not the first language are advised to consult with a technical language editor before submission.Conservation research ethics violated. Research was unnecessarily destructive, was conducted for the express purpose of causing harm/mortality (e.g., simulation of treatment or disturbance effects on survivorship), or violated ethics in the treatment and handling of animals. Where appropriate, authors must provide a statement and supporting documentation that research was approved by the authors' institutional animal care and use committee(s).Manuscripts that pass this first stage of editorial review are then subjected to a second stage of formal peer review. This involves evaluation of the manuscript by at least two specialists within the field of study, which may include one or more members of the editorial board. Beyond a critical assessment of the scientific content and overall presentation, referees are asked to evaluate the originality, likely impact and global relevance of the research. Referees make a recommendation to the handling editor, but note that it is ultimately the decision of the handling editor as to whether a manuscript is accepted for publication in Biological Conservation. Editor-in-Chief Dr. Vincent Devictor UMR CNRS-UM2 5554 cc065, Institut des Sciences de l'Evolution de Montpellier, Place Eugène Bataillon, 34090 Montpellier, France Email: vincent.devictor@univ-montp2.fr Editors Dr. Amanda Bates, University of Southampton, England, UK, Email: A.E.Bates@soton.ac.uk Dr. Richard Corlett, Richard Corlett Chinese Academy of Sciences (CAS), Menglun, Yunnan, China, Email: corlett@xtbg.org.cn AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 9 Dr. Graeme Cumming, James Cook University, Townsville, Queensland, Australia, Email: gscumming@gmail.com Dr. Liba Pejchar, Colorado State University, Colorado, USA, Email: Liba.Pejchar@colostate.edu Dr. Lian Pin Koh, University of Adelaide, Adelaide, South Australia, Australia, Email: lianpinkoh@gmail.com Dr. Rafael Loyola, Uniersidade Federal de Gois, Goinia, Gois, Brazil, Email: rdiasloyola@gmail.com Dr. Bea Maas, University of Vienna, Vienna, Austria, Email: beamaas@gmx.at Prof. Robin Pakeman, The James Hutton Institute, Aberdeen, Scotland, UK, Email: robin.pakeman@hutton.ac.uk Dr. Richard B. Primack, Boston University, Boston, Massachusetts, USA, Email: primack@bu.edu Dr. Tracey Regan, Arthur Rylah Institute for Environmental Research, Heidelberg, Victoria, Australia, Email: tregan@unimelb.edu.au Book Review Editor David Johns PO Box 725, McMinneville, OR 97218, Email: johnsd@pdx.edu PREPARATION NEW SUBMISSIONS Submission to this journal proceeds totally online and you will be guided stepwise through the creation and uploading of your files. The system automatically converts your files to a single PDF file, which is used in the peer-review process. As part of the Your Paper Your Way service, you may choose to submit your manuscript as a single file to be used in the refereeing process. This can be a PDF file or a Word document, in any format or lay- out that can be used by referees to evaluate your manuscript. It should contain high enough quality figures for refereeing. If you prefer to do so, you may still provide all or some of the source files at the initial submission. Please note that individual figure files larger than 10 MB must be uploaded separately. Please use correct, continuous line numbering and page numbering throughout the document. References There are no strict requirements on reference formatting at submission. References can be in any style or format as long as the style is consistent. Where applicable, author(s) name(s), journal title/ book title, chapter title/article title, year of publication, volume number/book chapter and the article number or pagination must be present. Use of DOI is highly encouraged. The reference style used by the journal will be applied to the accepted article by Elsevier at the proof stage. Note that missing data will be highlighted at proof stage for the author to correct. Formatting requirements There are no strict formatting requirements but all manuscripts must contain the essential elements needed to convey your manuscript, for example Abstract, Keywords, Introduction, Materials and Methods, Results, Conclusions, Artwork and Tables with Captions. If your article includes any Videos and/or other Supplementary material, this should be included in your initial submission for peer review purposes. Divide the article into clearly defined sections. Tables and Figures Please place legends above Tables and below Figures. They should follow the References at the end of the manuscript. AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 10 Peer review This journal operates a double blind review process. All contributions will be initially assessed by the editor for suitability for the journal. Papers deemed suitable are then typically sent to a minimum of two independent expert reviewers to assess the scientific quality of the paper. The Editor is responsible for the final decision regarding acceptance or rejection of articles. The Editor's decision is final. More information on types of peer review. Double-blind review This journal uses double-blind review, which means the identities of the authors are concealed from the reviewers, and vice versa. More information is available on our website. To facilitate this, please include the following separately: Title page (with author details): This should include the title, authors' names, affiliations, acknowledgements and any Declaration of Interest statement, and a complete address for the corresponding author including an e-mail address. Blinded manuscript (no author details): The main body of the paper (including the references, figures, tables and any acknowledgements) should not include any identifying information, such as the authors' names or affiliations. REVISED SUBMISSIONS Use of word processing software Please use correct, continuous line numbering and page numbering throughout the document. It is important that the file be saved in the native format of the word processor used. The text should be in single-column format. Keep the layout of the text as simple as possible. Most formatting codes will be removed and replaced on processing the article. In particular, do not use the word processor's options to justify text or to hyphenate words. However, do use bold face, italics, subscripts, superscripts etc. When preparing tables, if you are using a table grid, use only one grid for each individual table and not a grid for each row. If no grid is used, use tabs, not spaces, to align columns. The electronic text should be prepared in a way very similar to that of conventional manuscripts (see also the Guide to Publishing with Elsevier: https://www.elsevier.com/guidepublication). Note that source files of figures, tables and text graphics will be required whether or not you embed your figures in the text. See also the section on Electronic artwork. To avoid unnecessary errors you are strongly advised to use the 'spell-check' and 'grammar-check' functions of your word processor. Please use single spacing throughout the document. Use continuous line numbering throughout the document. Avoid full justification, i.e., do not use a constant right-hand margin. Ensure that each new paragraph is clearly indicated. Number every page of the manuscript, including the title page, references tables, etc. Present tables and figure legends on separate pages at the end of the manuscript. Layout and conventions must conform with those given in this guide to authors. Journal style has changed over time so do not use old issues as a guide. Number all pages consecutively. Italics are not to be used for expressions of Latin origin, for example, in vivo, et al., per se. Use decimal points (not commas); use a space for thousands (10 000 and above). Use of word processing software Regardless of the file format of the original submission, at revision you must provide us with an editable file of the entire article. Keep the layout of the text as simple as possible. Most formatting codes will be removed and replaced on processing the article. The electronic text should be prepared in a way very similar to that of conventional manuscripts (see also the Guide to Publishing with Elsevier). See also the section on Electronic artwork. To avoid unnecessary errors you are strongly advised to use the 'spell-check' and 'grammar-check' functions of your word processor. Cover letter Submission of a manuscript must be accompanied by a cover letter that includes the following statements or acknowledgements: The work is all original research carried out by the authors.All authors agree with the contents of the manuscript and its submission to the journal.No part of the research has been published in any form elsewhere, unless it is fully acknowledged in the manuscript. Authors should disclose how the research featured in the manuscript relates to any other manuscript of a similar nature that they have published, in press, submitted or will soon https://www.elsevier.com/reviewers/what-is-peer-review https://www.elsevier.com/reviewers/what-is-peer-review https://www.elsevier.com/reviewers/what-is-peer-review https://www.elsevier.com/authors/journal-authors/submit-your-paper https://www.elsevier.com/authors/journal-authors/submit-your-paper AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 11 submit to Biological Conservation or elsewhere.The manuscript is not being considered for publication elsewhere while it is being considered for publication in this journal.Any research in the paper not carried out by the authors is fully acknowledged in the manuscript.All sources of funding are acknowledged in the manuscript, and authors have declared any direct financial benefits that could result from publication.All appropriate ethics and other approvals were obtained for the research. Where appropriate, authors should state that their research protocols have been approved by an authorized animal care or ethics committee, and include a reference to the code of practice adopted for the reported experimentation or methodology. The Editor will take account of animal welfare issues and reserves the right not to publish, especially if the research involves protocols that are inconsistent with commonly accepted norms of animal research.Please include a short paragraph that describes the main finding of your paper, and its significance to the field of conservation biology.The authors should state in the cover letter if the paper in any form has previously been submitted to Biological Conservation. In that case the authors should specify the original manuscript number. Article structure Subdivision - numbered sections Divide your article into clearly defined and numbered sections. Subsections should be numbered 1.1 (then 1.1.1, 1.1.2, ...), 1.2, etc. (the abstract is not included in section numbering). Use this numbering also for internal cross-referencing: do not just refer to 'the text'. Any subsection may be given a brief heading. Each heading should appear on its own separate line. Introduction State the objectives of the work and provide an adequate background, avoiding a detailed literature survey or a summary of the results. Material and methods Provide sufficient details to allow the work to be reproduced by an independent researcher. Methods that are already published should be summarized, and indicated by a reference. If quoting directly from a previously published method, use quotation marks and also cite the source. Any modifications to existing methods should also be described. Theory/calculation A Theory section should extend, not repeat, the background to the article already dealt with in the Introduction and lay the foundation for further work. In contrast, a Calculation section represents a practical development from a theoretical basis. Results Results should be clear and concise. Discussion This should explore the significance of the results of the work, not repeat them. A combined Results and Discussion section is often appropriate. Avoid extensive citations and discussion of published literature. Conclusions The main conclusions of the study may be presented in a short Conclusions section, which may stand alone or form a subsection of a Discussion or Results and Discussion section. Glossary Please supply, as a separate list, the definitions of field-specific terms used in your article. Appendices If there is more than one appendix, they should be identified as A, B, etc. Formulae and equations in appendices should be given separate numbering: Eq. (A.1), Eq. (A.2), etc.; in a subsequent appendix, Eq. (B.1) and so on. Similarly for tables and figures: Table A.1; Fig. A.1, etc. Essential title page information • Title. Concise and informative, yet not overly general. If appropriate, include the species or ecosystem that was the subject of the study, or the location where the study was done. Titles are often used in information-retrieval systems. Avoid abbreviations and formulae where possible • Author names and affiliations. Where the family name may be ambiguous (e.g., a double name), please indicate this clearly. Present the authors' affiliation addresses (where the actual work was done) below the names. Indicate all affiliations with a lower-case superscript letter immediately after the author's name and in front of the appropriate address. Provide the full postal address of each affiliation, including the country name and, if available, the e-mail address of each author. AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 12 • Corresponding author. Clearly indicate who will handle correspondence at all stages of refereeing and publication, also post-publication. Ensure that phone numbers (with country and area code) are provided in addition to the e-mail address and the complete postal address. Contact details must be kept up to date by the corresponding author. • Present/permanent address. If an author has moved since the work described in the article was done, or was visiting at the time, a 'Present address' (or 'Permanent address') may be indicated as a footnote to that author's name. The address at which the author actually did the work must be retained as the main, affiliation address. Superscript Arabic numerals are used for such footnotes. Highlights Highlights are mandatory for this journal as they help increase the discoverability of your article via search engines. They consist of a short collection of bullet points that capture the novel results of your research as well as new methods that were used during the study (if any). Please have a look at the examples here: example Highlights. Highlights should be submitted in a separate editable file in the online submission system. Please use 'Highlights' in the file name and include 3 to 5 bullet points (maximum 85 characters, including spaces, per bullet point). Abstract A concise and factual abstract is required (maximum length of 250 words). The abstract should state briefly the purpose of the research, the methods used, the principal results and major conclusions. Please try to keep each sentence as specific as possible, and avoid such general statements as "The management implications of the results are discussed". An abstract is often presented separately from the article, so it must be able to stand alone. For this reason, References should be avoided, but if essential, they must be cited in full, without reference to the reference list. Also, non-standard or uncommon abbreviations should be avoided, but if essential they must be defined at their first mention in the abstract itself. Graphical abstract Although a graphical abstract is optional, its use is encouraged as it draws more attention to the online article. The graphical abstract should summarize the contents of the article in a concise, pictorial form designed to capture the attention of a wide readership. Graphical abstracts should be submitted as a separate file in the online submission system. Image size: Please provide an image with a minimum of 531 × 1328 pixels (h × w) or proportionally more. The image should be readable at a size of 5 × 13 cm using a regular screen resolution of 96 dpi. Preferred file types: TIFF, EPS, PDF or MS Office files. You can view Example Graphical Abstracts on our information site. Authors can make use of Elsevier's Illustration Services to ensure the best presentation of their images and in accordance with all technical requirements. Stereochemistry abstract For each important chiral compound you are requested to supply a stereochemistry abstract detailing structure, name, formula and all available stereochemical information for eventual incorporation into a database. An abstract for only one enantiomer per compound is required. Keywords Immediately after the abstract, provide a maximum of 6 keywords, using American spelling and avoiding general and plural terms and multiple concepts (avoid, for example, 'and', 'of'). Be sparing with abbreviations: only abbreviations firmly established in the field may be eligible. These keywords will be used for indexing purposes. Abbreviations Define abbreviations that are not standard in this field in a footnote to be placed on the first page of the article. Such abbreviations that are unavoidable in the abstract must be defined at their first mention there, as well as in the footnote. Ensure consistency of abbreviations throughout the article. Acknowledgements Collate acknowledgements in a separate section at the end of the article before the references and do not, therefore, include them on the title page, as a footnote to the title or otherwise. List here those individuals who provided help during the research (e.g., providing language help, writing assistance or proof reading the article, etc.). Formatting of funding sources List funding sources in this standard way to facilitate compliance to funder's requirements: https://www.elsevier.com/authors/journal-authors/highlights https://www.elsevier.com/authors/journal-authors/graphical-abstract https://webshop.elsevier.com/illustration-services/ AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 13 Funding: This work was supported by the National Institutes of Health [grant numbers xxxx, yyyy]; the Bill & Melinda Gates Foundation, Seattle, WA [grant number zzzz]; and the United States Institutes of Peace [grant number aaaa]. It is not necessary to include detailed descriptions on the program or type of grants and awards. When funding is from a block grant or other resources available to a university, college, or other research institution, submit the name of the institute or organization that provided the funding. If no funding has been provided for the research, please include the following sentence: This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Nomenclature and Units Follow internationally accepted rules and conventions: use the international system of units (SI) for all scientific and laboratory data. If other quantities are mentioned, give their equivalent in SI. Common names must be in lower-case except proper nouns. All common names must be followed by a scientific name in parentheses in italics. For example, bottlenose dolphin (Tursiops aduncus). Where scientific names are used in preference to common names they should be in italics and the genus should be reduced to the first letter after the first mention. For example, the first mention is given as Tursiops aduncus and subsequent mentions are given as T. aduncus. Math formulae Please submit math equations as editable text and not as images. Present simple formulae in line with normal text where possible and use the solidus (/) instead of a horizontal line for small fractional terms, e.g., X/Y. In principle, variables are to be presented in italics. Powers of e are often more conveniently denoted by exp. Number consecutively any equations that have to be displayed separately from the text (if referred to explicitly in the text). Footnotes Footnotes should be used sparingly. Number them consecutively throughout the article. Many word processors build footnotes into the text, and this feature may be used. Should this not be the case, indicate the position of footnotes in the text and present the footnotes themselves separately at the end of the article. Artwork Electronic artwork General points • Make sure you use uniform lettering and sizing of your original artwork. • Preferred fonts: Arial (or Helvetica), Times New Roman (or Times), Symbol, Courier. • Number the illustrations according to their sequence in the text. • Use a logical naming convention for your artwork files. • Indicate per figure if it is a single, 1.5 or 2-column fitting image. • For Word submissions only, you may still provide figures and their captions, and tables within a single file at the revision stage. • Please note that individual figure files larger than 10 MB must be provided in separate source files. A detailed guide on electronic artwork is available. You are urged to visit this site; some excerpts from the detailed information are given here. Formats Regardless of the application used, when your electronic artwork is finalized, please 'save as' or convert the images to one of the following formats (note the resolution requirements for line drawings, halftones, and line/halftone combinations given below): EPS (or PDF): Vector drawings. Embed the font or save the text as 'graphics'. TIFF (or JPG): Color or grayscale photographs (halftones): always use a minimum of 300 dpi. TIFF (or JPG): Bitmapped line drawings: use a minimum of 1000 dpi. TIFF (or JPG): Combinations bitmapped line/half-tone (color or grayscale): a minimum of 500 dpi is required. Please do not: • Supply files that are optimized for screen use (e.g., GIF, BMP, PICT, WPG); the resolution is too low. https://www.elsevier.com/authors/author-schemas/artwork-and-media-instructions AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 14 • Supply files that are too low in resolution. • Submit graphics that are disproportionately large for the content. Color artwork Please make sure that artwork files are in an acceptable format (TIFF (or JPEG), EPS (or PDF), or MS Office files) and with the correct resolution. If, together with your accepted article, you submit usable color figures then Elsevier will ensure, at no additional charge, that these figures will appear in color online (e.g., ScienceDirect and other sites) regardless of whether or not these illustrations are reproduced in color in the printed version. For color reproduction in print, you will receive information regarding the costs from Elsevier after receipt of your accepted article. Please indicate your preference for color: in print or online only. Further information on the preparation of electronic artwork. Figure captions Ensure that each illustration has a caption. A caption should comprise a brief title (not on the figure itself) and a description of the illustration. Keep text in the illustrations themselves to a minimum but explain all symbols and abbreviations used. Tables Please submit tables as editable text and not as images. Tables can be placed either next to the relevant text in the article, or on separate page(s) at the end. Number tables consecutively in accordance with their appearance in the text and place any table notes below the table body. Be sparing in the use of tables and ensure that the data presented in them do not duplicate results described elsewhere in the article. Please avoid using vertical rules and shading in table cells. References Citation in text Please ensure that every reference cited in the text is also present in the reference list (and vice versa). Any references cited in the abstract must be given in full. Unpublished results and personal communications are not recommended in the reference list, but may be mentioned in the text. If these references are included in the reference list they should follow the standard reference style of the journal and should include a substitution of the publication date with either 'Unpublished results' or 'Personal communication'. Citation of a reference as 'in press' implies that the item has been accepted for publication. Reference links Increased discoverability of research and high quality peer review are ensured by online links to the sources cited. In order to allow us to create links to abstracting and indexing services, such as Scopus, CrossRef and PubMed, please ensure that data provided in the references are correct. Please note that incorrect surnames, journal/book titles, publication year and pagination may prevent link creation. When copying references, please be careful as they may already contain errors. Use of the DOI is highly encouraged. A DOI is guaranteed never to change, so you can use it as a permanent link to any electronic article. An example of a citation using DOI for an article not yet in an issue is: VanDecar J.C., Russo R.M., James D.E., Ambeh W.B., Franke M. (2003). Aseismic continuation of the Lesser Antilles slab beneath northeastern Venezuela. Journal of Geophysical Research, https://doi.org/10.1029/2001JB000884. Please note the format of such citations should be in the same style as all other references in the paper. Web references As a minimum, the full URL should be given and the date when the reference was last accessed. Any further information, if known (DOI, author names, dates, reference to a source publication, etc.), should also be given. Web references can be listed separately (e.g., after the reference list) under a different heading if desired, or can be included in the reference list. Data references This journal encourages you to cite underlying or relevant datasets in your manuscript by citing them in your text and including a data reference in your Reference List. Data references should include the following elements: author name(s), dataset title, data repository, version (where available), year, and global persistent identifier. Add [dataset] immediately before the reference so we can properly identify it as a data reference. The [dataset] identifier will not appear in your published article. https://www.elsevier.com/authors/author-schemas/artwork-and-media-instructions https://www.elsevier.com/authors/author-schemas/artwork-and-media-instructions AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 15 References in a special issue Please ensure that the words 'this issue' are added to any references in the list (and any citations in the text) to other articles in the same Special Issue. Reference management software Most Elsevier journals have their reference template available in many of the most popular reference management software products. These include all products that support Citation Style Language styles, such as Mendeley. Using citation plug-ins from these products, authors only need to select the appropriate journal template when preparing their article, after which citations and bibliographies will be automatically formatted in the journal's style. If no template is yet available for this journal, please follow the format of the sample references and citations as shown in this Guide. If you use reference management software, please ensure that you remove all field codes before submitting the electronic manuscript. More information on how to remove field codes from different reference management software. Users of Mendeley Desktop can easily install the reference style for this journal by clicking the following link: http://open.mendeley.com/use-citation-style/biological-conservation When preparing your manuscript, you will then be able to select this style using the Mendeley plug- ins for Microsoft Word or LibreOffice. Reference formatting There are no strict requirements on reference formatting at submission. References can be in any style or format as long as the style is consistent. Where applicable, author(s) name(s), journal title/ book title, chapter title/article title, year of publication, volume number/book chapter and the article number or pagination must be present. Use of DOI is highly encouraged. The reference style used by the journal will be applied to the accepted article by Elsevier at the proof stage. Note that missing data will be highlighted at proof stage for the author to correct. If you do wish to format the references yourself they should be arranged according to the following examples: Reference style Text: All citations in the text should refer to: 1. Single author: the author's name (without initials, unless there is ambiguity) and the year of publication; 2. Two authors: both authors' names and the year of publication; 3. Three or more authors: first author's name followed by 'et al.' and the year of publication. Citations may be made directly (or parenthetically). Groups of references can be listed either first alphabetically, then chronologically, or vice versa. Examples: 'as demonstrated (Allan, 2000a, 2000b, 1999; Allan and Jones, 1999)…. Or, as demonstrated (Jones, 1999; Allan, 2000)… Kramer et al. (2010) have recently shown …' List: References should be arranged first alphabetically and then further sorted chronologically if necessary. More than one reference from the same author(s) in the same year must be identified by the letters 'a', 'b', 'c', etc., placed after the year of publication. Examples: Reference to a journal publication: Van der Geer, J., Hanraads, J.A.J., Lupton, R.A., 2010. The art of writing a scientific article. J. Sci. Commun. 163, 51–59. https://doi.org/10.1016/j.Sc.2010.00372. Reference to a journal publication with an article number: Van der Geer, J., Hanraads, J.A.J., Lupton, R.A., 2018. The art of writing a scientific article. Heliyon. 19, e00205. https://doi.org/10.1016/j.heliyon.2018.e00205. Reference to a book: Strunk Jr., W., White, E.B., 2000. The Elements of Style, fourth ed. Longman, New York. Reference to a chapter in an edited book: Mettam, G.R., Adams, L.B., 2009. How to prepare an electronic version of your article, in: Jones, B.S., Smith , R.Z. (Eds.), Introduction to the Electronic Age. E-Publishing Inc., New York, pp. 281–304. Reference to a website: Cancer Research UK, 1975. Cancer statistics reports for the UK. http://www.cancerresearchuk.org/ aboutcancer/statistics/cancerstatsreport/ (accessed 13 March 2003). Reference to a dataset: [dataset] Oguro, M., Imahiro, S., Saito, S., Nakashizuka, T., 2015. Mortality data for Japanese oak wilt disease and surrounding forest compositions. Mendeley Data, v1. https://doi.org/10.17632/ xwj98nb39r.1. http://citationstyles.org http://citationstyles.org http://www.mendeley.com/features/reference-manager https://service.elsevier.com/app/answers/detail/a_id/26093 https://service.elsevier.com/app/answers/detail/a_id/26093 AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 16 Video Elsevier accepts video material and animation sequences to support and enhance your scientific research. Authors who have video or animation files that they wish to submit with their article are strongly encouraged to include links to these within the body of the article. This can be done in the same way as a figure or table by referring to the video or animation content and noting in the body text where it should be placed. All submitted files should be properly labeled so that they directly relate to the video file's content. In order to ensure that your video or animation material is directly usable, please provide the file in one of our recommended file formats with a preferred maximum size of 150 MB per file, 1 GB in total. Video and animation files supplied will be published online in the electronic version of your article in Elsevier Web products, including ScienceDirect. Please supply 'stills' with your files: you can choose any frame from the video or animation or make a separate image. These will be used instead of standard icons and will personalize the link to your video data. For more detailed instructions please visit our video instruction pages. Note: since video and animation cannot be embedded in the print version of the journal, please provide text for both the electronic and the print version for the portions of the article that refer to this content. Data visualization Include interactive data visualizations in your publication and let your readers interact and engage more closely with your research. Follow the instructions here to find out about available data visualization options and how to include them with your article. Supplementary material Supplementary material such as applications, images and sound clips, can be published with your article to enhance it. Submitted supplementary items are published exactly as they are received (Excel or PowerPoint files will appear as such online). Please submit your material together with the article and supply a concise, descriptive caption for each supplementary file. If you wish to make changes to supplementary material during any stage of the process, please make sure to provide an updated file. Do not annotate any corrections on a previous version. Please switch off the 'Track Changes' option in Microsoft Office files as these will appear in the published version. The supplementary material should be cited as an online Appendix to the paper, usually in the Methods. If it contains several tables, images and/or figures, these should be cited as Table A1, Figure A1 and so on. Authors are strongly encouraged to make the data supporting their paper available to readers through an open-access data repository and/or as an Appendix to the paper. For more details on journal data policy see the paragraphs on Data Depositing and Linking to and depositing data at PANGAEA. Data Depositing Ideally, data should be freely available online through a specialist data centre that provides a permanent archive (repository) for the dataset, and may integrate the data with other datasets using international standards. Examples include PANGAEA, and GBIF and its major contributors such as OBIS and VertNet. Some Ocean Data Centres may also provide this service. Where such a data centre does not exist, we ask that the data be made freely available online from a permanent archive (repository). Where possible, it should follow international data standards. This may be an institutional repository for its staff. The data should be accompanied by sufficient information (metadata) for the reader to understand its composition and origins, and determine if it is fit for their purpose. In particular, the data should allow the results of the publication to be reproduced. Data being downloadable from departmental or personal websites is not regarded as permanently archived. Research data This journal encourages and enables you to share data that supports your research publication where appropriate, and enables you to interlink the data with your published articles. Research data refers to the results of observations or experimentation that validate research findings. To facilitate reproducibility and data reuse, this journal also encourages you to share your software, code, models, algorithms, protocols, methods and other useful materials related to the project. https://www.sciencedirect.com https://www.elsevier.com/authors/author-schemas/artwork-and-media-instructions https://www.elsevier.com/authors/author-resources/data-visualization AUTHOR INFORMATION PACK 22 Feb 2020 www.elsevier.com/locate/biocon 17 Below are a number of ways in which you can associate data with your article or make a statement about the availability of your data when submitting your manuscript. If you are sharing data in one of these ways, you are encouraged to cite the data in your manuscript and reference list. Please refer to the "References" section for more information about data citation. For more information on depositing, sharing and using research data and other relevant research materials, visit the research data page. Data linking If you have made your research data available in a data repository, you can link your article directly to the dataset. Elsevier collaborates with a number of repositories to link articles on ScienceDirect with relevant repositories, giving readers access to underlying data that gives them a better understanding of the research described. There are different ways to link your datasets to your article. When available, you can directly link your dataset to your article by providing the relevant information in the submission system. For more information, visit the database linking page. For supported data repositories a repository banner will automatically appear next to your published article on ScienceDirect. In addition, you can link to relevant data or entities through identifiers within the text of your manuscript, using the following format: Database: xxxx (e.g., TAIR: AT1G01020; CCDC: 734053; PDB: 1XFN). Mendeley Data This journal supports Mendeley Data, enabling you to deposit any research data (including raw and processed data, video, code, software, algorithms, protocols, and methods) associated with your manuscript in a free-to-use, open access repository. During the submission process, after uploading your manuscript, you will have the opportunity to upload your relevant datasets directly to Mendeley Data. The datasets will be listed and directly accessible to readers next to your published article online. For more information, visit the Mendeley Data for journals page. Data in Brief You have the option of converting any or all parts of your supplementary or additional raw data into one or multiple data articles, a new kind of article that houses and describes your data. Data articles ensure that your data is actively reviewed, curated, formatted, indexed, given a DOI and publicly available to all upon publication. You are encouraged to submit your article for Data in Brief as an additional item directly alongside the revised version of your manuscript. If your research article is accepted, your data article will automatically be transferred over to Data in Brief where it will be editorially reviewed and published in the open access data journal, Data in Brief. Please note an open access fee of 600 USD is payable for publication in Data in Brief. Full details can be found on the Data in Brief website. Please use this template to write your Data in Brief. MethodsX You have the option of converting relevant protocols and methods into one or multiple MethodsX articles, a new kind of article that describes the details of customized research methods. Many researchers spend a significant amount of time on developing methods to fit their specific needs or setting, but often without getting credit for this part of their work. MethodsX, an open access journal, now publishes this information in order to make it searchable, peer reviewed, citable and reproducible. Authors are encouraged to submit their MethodsX article as an additional item directly alongside the revised version of their manuscript. If your research article is accepted, your methods article will automatically be transferred over to MethodsX where it will be editorially reviewed. Please note an open access fee is payable for publication in MethodsX. Full details can be found on the MethodsX website. Please use this template to prepare your MethodsX article. 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