New Records of Batrachochytrium Dendrobatidis in the Atlantic Forest of Northeastern Brazil

NORTH-WESTERN JOURNAL OF ZOOLOGY 9 (1): 210-213 ©NwjZ, Oradea, Romania, 2013 Article No.: 132505 http://biozoojournals.3x.ro/nwjz/index.html

New records of Batrachochytrium dendrobatidis in the Atlantic forest of Northeastern Brazil

Barnagleison Silva LISBOA1,2,*, Jessika Maria de Moura NEVES3, Filipe Augusto Cavalcanti do NASCIMENTO², Leonora TAVARES-BASTOS3 and Tamí MOTT4

1. Programa de Pós-Graduação em Biologia Animal, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Av. Professor Nelson Chaves, s/n; Cidade Universitária, 50670-420, Recife, PE, Brazil. 2. Setor de Zoologia, Museu de História Natural, Universidade Federal de Alagoas, Av. Aristeu de Andrade, Farol, 57051-090, Maceió, AL, Brazil. 3. Laboratório de Histologia e Embriologia, Instituto de Ciências Biológicas e da Saúde, Universidade Federal de Alagoas, Av. Lourival Melo Mota, s/n; Tabuleiro, 57072-970, Maceió, AL, Brazil. 4. Setor de Biodiversidade e Ecologia, Instituto de Ciências Biológicas e da Saúde, Universidade Federal de Alagoas, Av. Lourival Melo Mota, s/n; Tabuleiro, 57072-970, Maceió, AL, Brazil. *Corresponding author, B.S. Lisboa, E-mail: [email protected]

Received: 30. August 2012 / Accepted: 05. February 2013 / Available online: 16. February 2013 / Printed: June 2013

Abstract. The pathogenic chytrid fungus Batrachochytrium dendrobatidis threatens the amphibian diversity at global scale. Three different strains of B. dendrobatidis have been already found in Brazil, the country with the greatest diversity of anurans worldwide, and it is widely spread in the Atlantic rainforest. Here, we report the occurrence of B. dendrobatidis for the state of Alagoas, the second record from northeastern Brazil. We describe for the first time B. dendrobatidis infection in Aplastodiscus sibilatus, a Brazilian tree frog species. The impact of B. dendrobatidis on populations of Brazilian anurans is unknown and information on spatial and taxonomic distribution of this fungus in Brazil is urgently needed for the development and implementation of amphibian conservation action plans.

Key words: Chytrid fungal pathogen, Aplastodiscus sibilatus, Atlantic rainforest, state of Alagoas.

The worldwide decline in amphibian populations al. 1998, Daszak et al. 2003, Skerratt et al. 2007, is one of the most striking features of the global Lips et al. 2008, Wake & Vredenburg 2008). Bd biodiversity crisis (Houlahan et al. 2000) although thrives in moist habitats such as tropical rainfor- untangling the multiple causes for this phenome- ests (Lips et al. 2008, Woodhams et al. 2008), bi- non has proved challenging (Collins & Storfer omes that are also noted for their high diversity of 2003). As with many other taxa, habitat loss has amphibians (Duellman 1999). driven many amphibian populations to extinction In Brazil, three Bd strains have been already (Stuart et al. 2004, Wake & Vredenburg 2008). genotyped (Schloegel et al. 2012, Vieira et al. 2012) These effects have been compounded by a range and 41 anuran species have been found infected of other factors such as pollution, climate change, by Bd (Carnaval et al. 2006, Toledo et al. 2006a, ultraviolet radiation, animal trade, invasive spe- Toledo et al. 2006b, Sluys et al. 2007, Gründler et cies and infectious diseases (Stuart et al. 2004, al. 2012, Vieira et al. 2012). So far, records of Bd in Young et al. 2004, Blackburn et al. 2010). Brazil have been almost exclusively restricted to The latest contemporary threats to amphibian south and southeastern portions of the Atlantic populations to be identified is the emergence of rainforest - there is a single record of Bd from the Chytridiomycosis, a fungal disease caused by in- northeast of the country (municipality of Jaqueira, fection of Batrachochytrium dendrobatidis (Bd) in state of Pernambuco; Carnaval et al. 2006) and an keratinized tissues of both adult amphibians and apparent distributional gap of ca. 1,700 km in the larvae (Berger et al. 1998, Longcore et al. 1999). Brazilian distribution of Bd. Here, we describe the This disease causes hyperkeratosis, hyperplasia of second record of B. dendrobatidis in the northeast of the skin and disturbs the skin osmoregulatory Brazil (and the first for Alagoas state) and report function and may lead to the host death (Rosen- for the first time susceptibility of Aplastodiscus sibi- blum et al. 2008, Voyles et al. 2009). Due to its latus to Bd infection. enormous geographical distribution, Bd is recog- On May 2012, tadpoles of Hypsiboas freicanecae (n=7) and nized as global threat to amphibian diversity and Aplastodiscus sibilatus (n=17) were collected with a dip net is thought to be the main cause of enigmatic de- at the bottom of a permanent stream (approximately 20 clines and extinctions of populations of amphibi- cm depth) at Mata da Bananeira, Estação Ecológica de ans in pristine and well-protected areas (Berger et Murici (ESEC Murici) (Murici, Alagoas state, 9°12'00” S Batrachochytrium dendrobatidis in Alagoas 211

Figure 1. Batrachochytrium dendrobatidis diagnosis by macroscopical (lack of keratin on the jaw sheaths) and histological assessments of tadpole mouthparts of Aplastodiscus sibilatus (A, C) and Hypsiboas freicanecae (B, D) collected in Murici (Alagoas, Brazil). Arrow indicate B. dendrobatidis sporangia (C, D).

35°52'00” W, DATUM WGS84; 551 m elev.). The tadpoles at Coleção Herpetológica do Museu de História were preserved in 10% neutral buffered formalin and Natural da Universidade Federal de Alagoas, Bra- analyzed under a stereomicroscope (Leica MZ6). All the zil (MUFAL 10779 for Hypsiboas freicanecae, and tadpoles were found to lack keratin (in different degrees) MUFAL 10780 for Aplastodiscus sibilatus). This is in their mouthparts. To confirm the presence of Bd, a tadpole of each species was subjected to histological assess- the first report of infection of the tree frog Aplasto- ment following the protocol of Berger et al. (1998). discus sibilatus by Bd. This report represents the first published re- Detailed histological analysis confirmed that the cord of Bd for the state of Alagoas and it extends collected tadpoles of H. freicanecae and A. sibilatus Bd distribution 65 km southeast from the north- were infected by Bd. In addition to the lack of ernmost record of Bd in Pernambuco state (Hypsi- keratin in oral structures observed during macro- boas freicanecae at Jaqueira municipality; Carnaval scopical examinations, we also observed a lack of et al. 2006) and 1,650 km north from the closest structural organization in the epithelium and the southern record in Rio de Janeiro state (Bokerman- presence of zoosparangia of Bd during histological nohyla circumdata at Petrópolis municipaliy; assessment (Fig. 1). All specimens were deposited Toledo et al. 2006a, Fig. 2).

212 B.S. Lisboa et al.

Figure 2. Distribution of Batrachochytrium dendrobatidis in Brazilian Atlantic rainforest. Dark circles indicate infected populations reported by Carnaval et al. 2006, Toledo et al. 2006a, Sluys et al. 2007, Gründler et al. 2012, and Vieira et al. 2012. Black square represents the new record from the present study. 1. São Francisco de Paula-RS (Toledo et al. 2006b), 2. Lauro Muller-SC (Toledo et al. 2006b), 3. São Bento do Sul-SC (Toledo et al. 2006b), 4. Morretes-PR (Vieira et al. 2012), 5. Apiaí-SP (Carnaval et al. 2006), 6. Rio Claro-SP (Toledo et al. 2006b), 7. Iporanga-SP (Schloegel et al. 2012), 8. Peruíbe-SP (Toledo et al. 2006b), 9. Jundiaí-SP (Vieira et al. 2012), 10. Santo André-SP (Toledo et al. 2006b), 11. Camanducaia-MG (Toledo et al. 2006a), 12. Bertioga-SP (Schloegel et al. 2012), 13. Caçapava-SP (Toledo et al. 2006b), 14. Ilhabela-SP (Gründler et al. 2012), 15. São Luiz do Paraitinga-SP (Toledo et al. 2006b), 16. Ubatuba-SP (Toledo et al. 2006b), 17. Itamonte- MG (Carnaval et al. 2006), 18. Itatiaia-RJ (Sluys et al. 2007), 19. Nova Iguaçu-RJ (Carnaval et al. 2006), 20. Petrópolis-RJ (Toledo et al. 2006b), 21. Jaqueira-PE (Carnaval et al. 2006).

The presence of Bd in ESEC Murici, a State and in 18 of the 27 federal states (Carnaval et al. Park of ca. 60 km2, should be of particular concern 2005, Carnaval et al. 2006, Toledo et al. 2006a, because this area represents the largest rainforest Toledo et al. 2006b, Sluys et al. 2007, Gründler et fragment of Alagoas state (Assis 2000), and con- al. 2012, Vieira et al. 2012). This is very worrying, tains several poorly known anuran species such as as in a recent work, Schloegel et al. (2012) identi- Crossodactylus dantei, Phyllodytes gyrinaethes, Phy- fied a novel lineage of Bd (Bd-Brazil) in the Atlan- salaemus caete, Scinax muriciensis and Agalychnis tic rainforest of Brazil that is associated with the granulosa. The latter species is considered threat- global trade in bullfrogs, is capable of sexual re- ened according to the Red List of Brazilian fauna production, and is also detected on bullfrogs in- (Haddad 2008). troduced into Japan. Given the small sample size Batrachochytrium dendrobatidis has been found of this report, much more extensive sampling will in more than 200 species worldwide (Skerratt et al. be required to provide sufficiently detailed taxo- 2007). In Brazil, Bd has been there since at least nomic, spatial and genetic information about Bd in 1981 (Toledo et al. 2006a); it appears widespread northeastern Brazil to allow strategic conservation in the highly fragmented Atlantic rainforest bi- planning. ome, it has already been found in over 40 species Batrachochytrium dendrobatidis in Alagoas 213

Acknowledgments. We are grateful to L. Araujo, J. Neto, Lips, K.R., Diffendorfer, J., Mendelson, J.R., Sears, M.W. (2008): I. Tiburcio and N. Melo for assistance in fieldwork; R. J. Riding the wave: reconciling the roles of disease and climate Ladle made valuable suggestions on a previous version of change in amphibian declines. Plos Biology 6: 0441-0454. Longcore, J.E., Pessier, A.P., Nichols, D.K. (1999): Batrachochytrium the manuscript; L. F. Toledo confirmed histological dendrobatidis gen. et sp. nov., a chytrid pathogenic to diagnosis of Bd and provided the geographic coordinates amphibians. Mycologia 91: 219-227. of all localities where Bd has been found in Brazil; N. Rosenblum, E.B., Stajich, J., Maddox, N., Eisen, M.B. (2008): Global Fabré and M. Souza in permitting the use of their Leica gene expression profiles of life stages of the deadly amphibian stereomicroscope for multi-focus photography; S. pathogen Batrachochytrium dendrobatidis. Proceedings of the National Academy of Sciences of the United States of America Smaniotto in permitting the use of their Nikon 105: 17034-17039. microscope; ICMBio/SISBIO collection permits (#33507- Schloegel, L.M., Ferreira, C.M., James, T.Y., Hipolito, M., Longcore, 1). BSL thanks CNPq for scholar grant (Process J.E., Hyatt, A.D., Yabsley, M., Martins, A.M.C.R.P.F., Mazzoni, 132436/2012-4). R., Davies, A.J., Daszak, P. (2010). The North American bullfrog as a reservoir for the spread of Batrachochytrium dendrobatidis in Brazil. Animal Conservation 13: 53-61. Schloegel, L.M., Toledo, L.F., Longcore, J.E., Greenspan, S.E., Vieira,

C.A., Lee, M., Zhao, S., Wangen, C., Ferreira, C.M., Hipolito, M., References Davies, A.J., Cuomo, C.A., Daszak, P., James, T.Y. (2012): Novel,

panzootic and hybrid genotypes of amphibian chytridiomycosis Assis, J.S. (2000): Biogeografia e Conservação da Biodiversidade - associated with the bullfrog trade. Molecular Ecology 21(21): Projeções para Alagoas. Catavento, Maceió-São Paulo. 5162-5177. Berger, L., Speare, R., Daszak, P., Green, D.E., Cunningham, A.A., Skerratt, L.F., Berger, L., Speare, R., Cashins, S., McDonald, K.R., Goggin, C.L., Slocombe, R., Ragan, M.A., Hyatt, A.D., Phillott, A.D., Hines, H.B., Kenyon, N. (2007): Spread of McDonald, K.R., Hines, H.B., Lips, K.R., Marantelli, G., Parkes, chytridiomycosis has caused rapid global decline and extinction H. (1998): Chytridiomycosis causes amphibian mortality of frogs. EcoHealth 4: 125-134. associated with population declines in the rain forests of Sluys, M.V., Ferreria, T.K., Lamarão, F.R.M., Moraes, M.O. (2007): Australia and Central America. Proceedings of the National Batrachochytrium dendrobatidis infects Melanoprhyniscus moreirae Academy of Sciences of the United States of America 95: 9031- (Anura Bufonidae) from an Atlantic Rainforest area (Itatiaia), 9036. southeastern Brazil. Froglog 84: 6-7. Blackburn, D.C., Evans, B.J., Pessier, A.P., Vredenburg, V.T. (2010): Stuart, S.N., Chanson, J.S., Cox, N.A., Young, B.E., Rodrigues, An enigmatic mortality event in the only population of the A.S.L., Fischman, D.L., Waller, R.W. (2004): Status and trends of Critically Endangered Cameroonian frog Xenopus longipes. amphibian declines and extinctions worldwide. Science 306: African Journal of Herpetology 59: 111-122. 1783-1786. Carnaval, A.C.O.Q., Toledo, L.F., Haddad, C.F.B., Britto, F.B. (2005): Toledo, L.F., Britto, F.B., Araújo, O.G.S., Giasson, L.M.O., Haddad, Chytrid fungus infects high-altitude stream-dwelling Hylodes C.F.B. (2006a): The occurrence of Batrachochytrium dendrobatidis magalhaesi (Leptodactylidae) in the Brazilian Atlantic Rainforest. in Brazil and inclusion of 17 new cases. South American Journal Froglog 70: Art. #04, pp. 3-4. of Herpetology 1: 185-191. Carnaval, A.C.O.Q., Puschendorf, R., Peixoto, O.L., Verdade, V.K., Toledo, L.F., Haddad, C.F.B., Carnaval, A.O.C.Q., Britto, F.B. Rodrigues, M.T. (2006): Amphibian chytrid fungus broadly (2006b): A Brazilian anuran (Hylodes magalhaesi: distributed in the Brazilian Atlantic Rain Forest. EcoHealth 3: Leptodactylidae) infected by Batrachochytrium dendrobatidis: a 41-48. conservation concern. Amphibian and Reptile Conservation 4: Collins, J.P., Storfer, A. (2003): Global amphibian declines: sorting Art. #02, pp. 17-21. the hypotheses. Diversity and Distributions 9: 89-98. Vieira, C. A., Almeida, C. H. L. N., Lambertini, C., Leite, D.S., Daszak, P., Cunningham, A.A., Hyatt, A.D. (2003): Infectious Toledo, L.F. (2012): First Record of Batrachochytrium dendrobatidis disease and amphibian population declines. Diversity and in Paraná, Brazil 43: 93-94. Distributions 9: 141-150. Vieira, C.A., Toledo, L.F., Longcore, J.E., Longcore, J.E. (2013): Body Duellman, W.E. (1999): Patterns of distribution of amphibians: A length of Hylodes ornatus and Lithobates catesbeianus tadpoles, global perspective. The John Hopkins University Press, depigmentation of mouthparts, and presence of Baltimore & London. Batrachochytrium dendrobatidis are related. Brazilian Journal of Fisher, M.C., Garner, T.W.J., Walker, S.F. (2009): The global Biology 73: in press. emergence of Batrachochytrium dendrobatidis in space, time, and Voyles, J., Young, S., Berger, L., Campbell, C., Voyles, W.F., host. Annual Review of Microbiology 63: 291-310. Dinudom, A., Cook, D., Webb, R., Alford, R.A., Skerratt, L.F., Garner, T.W.J., Perkins, M.W., Govindarajulu, P., Seglie, D., Speare, R. (2009): Pathogenesis of chytridiomycosis, a cause of Walker, S., Cunningham, A.A., Fisher, M.C. (2006). The catastrophic amphibian declines. Science 326: 582-585. emerging amphibian pathogen Batrachochytrium dendrobatidis Wake, D.B., Vredenburg, V.T. (2008): Are we in the midst of the globally infects introduced populations of the North American sixth mass extinction? A view from the world of amphibians. bullfrog, Rana catesbeiana. Biology letters 2: 455-459. Proceedings of the National Academy of Sciences of the United Gründler, C.M., Toledo, L.F., Parra-Olea, G., Haddad, C.F.B., States of America 105: 11466-11473. Giasson, L.O.M., Sawaya, R.J., Prado, C.P.A., Araujo, O.G.S., Woodhams, D.C., Kilburn, V.L., Reinert, L.K., Voyles, J., Medina, Zara, F.J., Centeno, F.C., Zamudio, K. R. (2012): Interaction D., Ibáñez, R., Hyatt, A.D., Boyle, D.G., Pask, J.D., Green, D.M., between breeding habitat and elevation affects prevalence but Rollins-Smith, L.A. (2008): Chytridiomycosis and amphibian not infection intensity of Batrachochytrium dendrobatidis in population declines continue to spread eastward in Panama. Brazilian anuran assemblages. Diseases of Aquatic Organisms EcoHealth 5: 268-274. 97: 173-184. Young, B.E., Stuart, S.N., Chanson, J.S., Cox, N.A., Boucher, T.M. Haddad, C.F.B. (2008): Anfíbios. pp. 291-302. In: Machado, A.B.M., (2004): Disappearingjewels: The status of New World Drummond, G.M., Paglia, A.P. (eds.), Lista da Fauna Brasileira amphibians. NatureServe, Arlington, EUA. Ameaçada de Extinção. Biodiversitas, Belo Horizonte.

Houlahan, J.E., Findlay, C.S., Schmidt, B.R., Meyer, A.H., Kuzmin,

S.L. (2000): Quantitative evidence for global amphibian population declines. Nature 404: 752-755.