SciELO - Scientific Electronic Library Online

 
vol.57 issue2IDENTIFICATION OF Leishmania infantum IN PUERTO IGUAZÚ, MISIONES, ARGENTINANEW WILDLIFE HOSTS OF Leptospira interrogans IN CAMPECHE, MEXICO author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand

Journal

Article

  • text new page (beta)
  • English (pdf)
  • Article in xml format
  • How to cite this article
  • SciELO Analytics
  • Curriculum ScienTI
  • Automatic translation

Indicators

Related links

Share


Revista do Instituto de Medicina Tropical de São Paulo

Print version ISSN 0036-4665

Rev. Inst. Med. trop. S. Paulo vol.57 no.2 São Paulo Mar./Apr. 2015

https://doi.org/10.1590/S0036-46652015000200014 

BRIEF COMMUNICATIONS

DETECTION OF Leptospira spp. AND Brucella abortus ANTIBODIES IN FREE-LIVING JAGUARS (Panthera onca) IN TWO PROTECTED AREAS OF NORTHERN PANTANAL, BRAZIL

Detecção de anticorpos para Leptospira spp. e Brucella abortus em onças-pintadas (Panthera onca) de vida livre em duas áreas protegidas no Pantanal Norte, Brasil

Selma Samiko Miyazaki ONUMA(1)  (2) 

Daniel Luis Zanella KANTEK(1) 

Peter Gransden CRAWSHAW JÚNIOR(3) 

Ronaldo Gonçalves MORATO(3) 

Joares Adenilson MAY-JÚNIOR(4) 

Zenaide Maria de MORAIS(5) 

José Soares FERREIRA NETO(5) 

Daniel Moura de AGUIAR(1) 

(1)Estação Ecológica de Taiamã, Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio), Av. Getúlio Vargas, s/n, 78200-000 Cáceres, Mato Grosso, Brazil. E-mails: selma.onuma@icmbio.gov.br, daniel.kantek@icmbio.gov.br, danmoura@ufmt.br

(2)Laboratório de Virologia e Rickettsioses, Hospital Veterinário, Universidade Federal de Mato Grosso, Av. Fernando Correa da Costa 2367, 78060-900 Cuiabá, Mato Grosso. Brazil.

(3)Centro Nacional de Pesquisa e Conservação de Mamíferos Carnívoros, Instituto Chico Mendes de Conservação da Biodiversidade (CENAP/ICMBio), 12952-011 Atibaia, São Paulo, Brazil. E-mail: ronaldo.morato@icmbio.gov.br

(4)Instituto Pró-Carnívoros, 12945-010 Atibaia, São Paulo, Brazil. Universidade do Sul de Santa Catarina (UNISUL), 12952-011 Santa Catarina, SC, Brazil. E-mail: joares.may@ig.com.br

(5)Departamento de Medicina Veterinária Preventiva e Saúde Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, 05508-270 São Paulo, SP, Brazil. E-mails: zenaide@usp.br, jsoares@vps.fmvz.usp.br


ABSTRACT

 This study aimed to assess the exposure of free-living jaguars (Panthera onca) to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT). The Rose Bengal test was applied for B. abortus antibodies. Two (18.2%) jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01). All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection.

Key words: Zoonotic diseases; Leptospira spp.; B. abortus ; Serology; Panthera onca

RESUMO

Este estudo teve como objetivo avaliar a exposição de onças-pintadas de vida livre (Panthera onca) para Leptospira spp. e Brucella abortus em duas unidades de conservação no Pantanal de Mato Grosso, Brasil. A presença de anticorpos em amostras de sangue de onze onças foi investigada utilizando antígenos autóctones isoladas no Brasil adicionais a coleção de antígenos de referência aplicada usualmente ao diagnóstico da leptospirose pelo teste de soroaglutinação microscópica (MAT). Para os anticorpos de B. abortus, foi utilizado o teste de Rosa Bengala. Duas onças-pintadas (18,2%) foram reagentes para Leptospira spp. e o sorotipo considerado como o mais provável pela infecção em ambos os animais foi um isolado brasileiro do sorovar Canicola (L01). Todas as onças-pintadas foram soronegativas para B. abortus. Estes dados indicam que a inclusão de antígenos autóctones em estudos sorológicos pode aumentar significativamente o número de animais reativos, assim como modificar a caracterização do sorotipo mais prevalente.

INTRODUCTION

The jaguar (Panthera onca) is the largest felid in the Americas, where retaliation from ranchers due to livestock predation, illegal hunting tourism activity, and habitat loss associated with agricultural expansion are threats to the species in the Pantanal biome, an important area for jaguar conservation in the long-term4.

Leptospirosis is a zoonosis of worldwide distribution and global importance1. The incidence of human infection is higher in the tropics where conditions for its transmission are favorable, but the disease occurs in both industrialized and developing countries2. The genus Leptospira is divided into 20 species based on DNA hybridization studies2, and these 20 species are classified into more than 280 serovars, according to their antigenic relatedness5, which affect various vertebrate hosts and remain in the environment by a dynamic process through a variety of domestic and wild animals. Leptospires are shed in the urine of carrier animals and the transmission is strongly affected by environmental conditions2. In Brazil, serological surveys have shown exposure to Leptospira spp. in various captivity and free-living wild species, of which the serovars Castellonis, Hardjo10, and Copenhageni15 were the most likely to cause infection in captive jaguars. Pomona was the most prevalent serovar found in free-living sampled jaguars7 , 21. In addition to the death of a female puma in Rio de Janeiro's zoo which showed clinical signs of leptospirosis and titers ≥ 400 to serovar Pomona by MAT15, the high titers found in free-living neotropical felids in the same biome studied12 suggest that Lepstospira spp. exposure may affect the conservation of wild felids. Due to the fact that transmission occurs mainly in wet environments and that wild animals are relevant in leptospirosis epidemiology, studies are necessary in the Pantanal region to clarify the potential impact of Leptospira spp. exposure on wild populations.

The occurrence of brucellosis in humans is highly dependent on the occurrence of the disease in animals' reservoirs, including wildlife8. The main clinical signs of Brucella abortus in wild mammals are abortion, orchitis, epididymitis and infertility24. In Brazil, antibodies against B. abortus have been detected in free-living and captive white-lipped and collared peccaries11 , 13 , 18 , 21, in free-living and captive maned wolves (Chrysocyon brachyurus)16, in a free-living jaguar in the Atlantic Forest21 and in another in the Emas National Park12. The present study aimed to detect antibodies to Leptospira spp. and B. abortus in jaguars from two conservation units in the Pantanal region, Brazil.

MATERIAL AND METHODS

The studied areas comprised two federal conservation units (Taiamã Ecological Station - 16° 50' 34.31" S, 57° 35' 03.70" W, and Pantanal Matogrossense National Park - 17° 50'47. 33" S, 57° 24'12.67" W) in the Pantanal of Mato Grosso, considered one of the largest floodplains in the world. Annual rainfall ranges from 1,200 to 1,300 mm across the region, defining dry and rainy seasons with seasonal fluctuations in water level, which have great influence on ecological processes17. The average annual temperature is 25 °C3.

Eleven jaguars were captured between July 2010 and November 2012, under license granted by the Authorization System and Biodiversity Information - SISBIO, numbers: 30896-1 and 18699-1, immobilized with a combination of tiletamine and zolazepam (Zoletil 100®, Virbac SA, Carros-Cedex, France) and fitted with radio-collars. After clinical examination and collection of biological samples, all animals were released at the same site at which they were captured. Sera blood samples were frozen and stored at -20 °C until testing and analysis at the Biological Samples' Bank of National Research Center for the Conservation of Carnivorous Mammals of Brazil (CENAP/ ICMBio).

Serum samples were examined for different leptospiral antibodies by Microscopic Agglutination Test (MAT)6 with the cut off 1:100 dilution against the following pathogenic serovars: Australis, Bratislava, Autumnalis, Butembo, Castellonis, Bataviae, Canicola, Whiticombi, Cynopteri, Grippotyphosa, Hebdomadis, Copenhageni, Icterohaemorrhagiae, Mini, Javanica, Panama, Pomona, Pyrogenes, Hardjo, Wolffi, Shermani, Tarassovi, Andamana, Patoc and Sentot, which were cultivated in modified EMJH medium. In addition to the reference strains, eleven Brazilian isolates of Leptospira spp. were used in this study: Brasiliense serovar isolated from Didelphus marsupialis (Strain 4B), Pomona serovar isolated from domestic Sus scrofa (Strain M7/87), Guaricura serovar isolated from Bubalus bubalis (Strain M4/98), Copenhageni serovar isolated from Rattus norvegicus (Strain M9/99), Canicola serovar isolated from Canis familiars (Strain L01), Canicola serovar isolated from domestic Sus scrofa (Strain L04), Canicola serovar isolated from Bos Taurus (Strain L014), Bananal serovar isolated from Hydrochaeris hidrochaeris (Strain 2A CAP), Bananal serovar isolated from Hydrochaeris hidrochaeris (Strain 21A CAP), Pomona serovar isolated from domestic Sus Scrofa (Strain Gr6) and M110/06 isolated from Cerdocyon thous (probably a new species). The positive sera were titrated by testing serial twofold serum dilutions and the reciprocal of the highest serum dilution that showed 50% agglutinated leptospira was defined as the serum titer20.

For brucellosis, serum samples were examined by the Rose Bengal test (RBT) for screening and 2-mercaptoethanol test (2-ME) as a confirmatory test14. The antigen used was an inactive suspension of B. abortus 1119-3 produced by the Institute of Technology of Paraná, Brazil.

RESULTS

All animals were considered adults based on tooth wear and color, ranging from four to 10 years old. Only two (18.2%) jaguars tested were seroreactive for Leptospira spp. antigen by MAT, one from each conservation unit of this study. The serovar considered as most infective in both animals was a Brazilian isolated antigen, serovar Canicola (L01) with titers = 3200. One of the seropositive animals reacted only to serovars of Brazilian isolate antigens (Canicola L01, T = 3200. Canicola L04, T = 800; Canicola L014, T = 400), and the other animal showed a low titer for serovar Copenhageni (10A), the only reaction for antigen of the reference collection (Canicola L01, T = 3200; Canicola L04, T = 400; Canicola L014, T = 400; Copenhageni 10A, T = 200: Copenhageni M9/99, T = 200). Both animals were in good overall health at the time of capture and no clinical signs were correlated with the infections dealt with in the present paper. All eleven jaguar serum samples were negative for B. abortus antigen by RBT.

DISCUSSION

Despite the absence of clinical signs at the time of capture, high antibody titers to serovar Canicola (L01) isolated from Canis familiaris in the state of Paraná, southern Brazil, were detected in two jaguars highlighting the importance of using local antigens in serological surveys, usually carried out only with collection of reference antigens performed by MAT. This high titer (T = 3200) also suggests a recent or frequent contact with this or a closely related agent, which probably circulates in these two regions of the northern Pantanal, despite the low frequency of positive animals found in this study (18.2%). Although the specificity of MAT is good, there is significant serological cross-reactivity among serovars that may result in an equal or even higher antibody titer19. Therefore, serological tests may suggest, but not definitively identify, the infecting serovar, and isolation of the agent and molecular analysis should be required.

Serological surveys are commonly performed in studies of wildlife exposure to pathogens in South America to determine whether wild animals have been exposed to an antigen, due to the concern of disease transmission across the interface between wildlife and domestic animals. However, local Brazilian antigens have rarely been tested in studies involving free-living species in serological investigations for leptospiral infections. VIEIRA et al. (2013)23 did not detect seropositivity to serovar Canicola (L01) using local strains in a study of exposure to Leptospira spp. in wild mammals from the southern Pantanal of Mato Grosso do Sul, and the only other antigen isolated in Brazil of serovar Canicola (L014) was detected in Cerdocyon thous. In the present study, the results showed high antibody titers for serovar Canicola, of which the main natural reservoir is the dog, suggesting an occasional contact between jaguars and domestic dogs. This is a concern for the conservation of wild carnivores and requires further investigations in the Pantanal region. Furthermore, the inclusion of autochthonous antigens in serological inquiries should be considered because they can significantly increase the number of reactive animals, as well as modify the epidemiological profile of infections, likewise observed in a study conducted on cattle by SARMENTO et al. (2012)22.

The serological method used to detect B. abortus antibodies in the present study is recommended by the Brazilian Department of Livestock Health14 and is also performed in serological surveys for brucellosis in wildlife7 , 21. FURTADO (2010)7 reported the contact of a jaguar with Brucella sp. in the Pantanal biome, suggesting that predation of cattle infected with B. abortus may explain seroconversion. In the present work, although all samples were negative for antibodies against B. abortus, zoonotic diseases included in sanitary control programs need to be further investigated in wildlife to assist decision-makers in the development of effective action plans. The negativity of exposure to B. abortus in jaguars from these two conservation units suggests a low level of environmental anthropogenic alteration, commonly related to livestock areas, with consequent predation of cattle. However, the validation of diagnostic techniques, such as serology, requires careful analysis of the isolation of Brucella agent, especially when it comes to wildlife9, in order to determine the possible transmission chain of the disease and the role of certain species in the maintenance of the agent in the environment.

Despite the low number of reactive animals for Leptospira antigens and the absence of individuals positive for B. abortus antibodies in the present study, more extensive investigations are necessary to determine the likelihood of the impact of infection by these pathogens on the health and reproductive parameters of wild populations. These data are important for the development of management plans of protected areas, as well as for an evaluation of the role played by this species in the epidemiological cycle of this important zoonotic disease.

Preventive measures for Leptospira spp. infection, such as water treatment for consumption and chemoprophylaxis, can also be recommended for people who visit these areas for professional reasons or for recreational activities.

Acknowledgements

ACKNOWLEDGMENTS

The authors thank the researchers of the Virology and Rickettsiosis Laboratory (UFMT) and Bacterial Zoonosis (USP) for technical support. This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq - research grant and scholarship to DMA). The authors also thank the National Research Center for the Conservation of Carnivorous Mammals (CENAP/ ICMBio) for financial and technical support in the capture of animals.

REFERENCES

1.  Acha PN, Szifres B. Zoonosis y enfermidades transmisibles comuns al hombre y a los animales. Bacteriosis y micosis. 3rd ed. Washington: Organización Panamericana de La Salud; 2001. v. 1. (Publ. Cient. Tecn. nº 580). [ Links ]

2.  Bharti AR, Nally JE, Ricaldi JN, Mathias MA, Diaz MM, Lovett MA, et al. Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis. 2003;3:757-71. [ Links ]

Calheiros DF, Fonseca-Júnior WC. Perspectivas de estudos ecológicos sobre o Pantanal. Corumbá: EMBRAPA-CPAP. 1996. (EMBRAPA-CPAP Documento 18). [ Links ]

Cavalcanti SMC, Azevedo FCC, Tomás WM, Boulhosa RLP, Crawshaw PG Jr. The status of the jaguar in the Pantanal. CATnews. 2012(Special Issue 7):29-34. [ Links ]

5.  Cerqueira GM, Picardeau M. A century of Leptospira strain typing. Infect Genet Evol. 2009;9:760-8. [ Links ]

6.  Faine S, Adler B, Boein C, Perolat P. Leptospira and leptospirosis. 2nd ed. Melbourne: MedSci; 2000. [ Links ]

Furtado MM. Estudo epidemiológico de patógenos circulantes nas populações de onça-pintada e animais domésticos em áreas preservadas de três biomas brasileiros: Cerrado, Pantanal e Amazônia. [Tese]. São Paulo: Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia; 2010. [ Links ]

Godfroid J. Brucellosis in wildlife. Rev Sci Tech. 2002;21:277-86. [ Links ]

Godfroid J, Nielsen K, Saegerman C. Diagnosis of brucellosis in livestock and wildlife. Croat Med J. 2010;51:296-305. [ Links ]

10.  Guerra-Neto G, Girio RJS, Andrade TM, Koproski LP, Moraes W, Santos LC. Ocorrência de anticorpos contra Leptospira spp. em felídeos neotropicais pertencentes ao criadouro de animais silvestres da Itaipu binacional e ao zoológico municipal Bosque Guarani, Foz do Iguaçu Estado do Paraná. Ars Vet. 2004;20:75-80. [ Links ]

Ito FH, Vasconcellos AS, Bernardi F, Nascimento AA, Labruna MB, Arantes IG. Evidência sorológica de brucelose e leptospirose e parasitismo por ixodídeos em animais silvestres do pantanal sul-matogrossense. Ars Vet. 1998;14:301-10. [ Links ]

12.  Jorge RPS, Ferreira F, Ferreira Neto JS, Vasconcellos SA, Lima E, Morais ZM, et al. Exposure of free-ranging wild carnivores, horses and domestic dogs to Leptospira spp in the northern Pantanal, Brazil. Mem Inst Oswaldo Cruz. 2011;106:441-4. [ Links ]

13.  Kashivakura CK, Furtado MM, Jacomo ATA, Marvulo MF, Silva JCR, Suero D, et al. Brucelose em queixadas (Tayassu pecari) de vida livre da região do Parque Nacional das Emas. In: 25º Congresso Brasileiro de Zoologia; 2004; Brasília. Anais. Brasília: Sociedade Brasileira de Zoologia; 2004. p. 217-8. [ Links ]

14.  Lage AP, Roxo E, Muller EE, Poester FP, Cavalléro JCM, Ferreira Neto JS, et al. Programa nacional de controle e erradicação da brucelose e da tuberculose animal. Brasília: MAPA/SDA/DSA; 2006. [ Links ]

15.  Lilenbaum W, Monteiro RV, Albuquerque CE, Ristow P, Fraguas S, Cardoso VS, et al. Leptospiral antibodies in wild felines from Rio de Janeiro Zoo, Brazil. Vet J. 2004;168:191-3. [ Links ]

16.  Maia OM, Veloso I, Viana FJ, Guimarães PHS, Leite RM, Lage AP. Avaliação sorológica para leptospirose e brucelose em lobos guarás (Chrysocyon brachyurus Illiger 1811) provenientes da natureza e de cativeiro. In: 24º Congresso Brasileiro da Sociedade de Zoológicos do Brasil, 5º Encontro Internacional de Zoológicos; Belo Horizonte. Anais. Belo Horizonte; 2000. p. 43. [ Links ]

Mamede SB, Alho CJR. Response of wild mammals to seasonal shrinking-and-expansion of habitats due to flooding regime of the Pantanal, Brazil. Braz J Biol. 2006;66:991-8. [ Links ]

18.  Mayor P, Le Pendu Y, Guimarães BDA, Silva JV, Tavares HL, Tello MA, et al. A health evaluation in a colony of captive collared peccaries (Tayassu tacaju) in the Eastern Amazon. Res Vet Sci. 2006;81:246-53. [ Links ]

Modrić Z, Herceg M, Župančić Ž, Bambir S, Hahn V, Ramadan P. Leptospiroza pasa u Zagrebu i okolici uzrokovana serološkim tipom icterohaemorrhagiae. Vet Arhiv. 1985;55:93-102. [ Links ]

Myers D. Leptospirosis: manual de métodos para el diagnostico de laboratório. Buenos Aires: Centro Panamericano de Zoonoses; 1985. (Nota técnica 30). [ Links ]

Nava AFD. Espécies sentinelas para a Mata Atlântica: as consequências epidemiológicas da fragmentação florestal no Pontal do Paranapanema, São Paulo. [Tese]. São Paulo: Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia; 2008. [ Links ]

22.  Sarmento AMC, Azevedo SS, Morais ZM, Souza GO, Oliveira FCS, Gonçalves AP, et al. Emprego de estirpes Leptospira spp. isoladas no Brasil na microtécnica de soroaglutinação microscópica aplicada ao diagnóstico da leptospirose em rebanhos bovinos de oito estados brasileiros. Pesq Vet Bras. 2012;32:601-6. [ Links ]

23.  Vieira AS, Rosinha GMS, Vasconcellos SA, Morais ZM, Viana RC, Oliveira CE, et al. Identificação de mamíferos silvestres do Pantanal Sul Mato-grossense portadores de Leptospira spp. Ciênc Anim Bras. 2013;14:373-80. [ Links ]

24.  Williams ES, Baker IK. Infectious diseases of wild mammals. 3rd ed. Ames: Iowa State University Press; 2001. p. 323-31. [ Links ]

Received: April 11, 2014; Accepted: July 10, 2014

Correspondence to: Selma Samiko Miyazaki Onuma. E-mail: selma.onuma@icmbio.gov.br

Creative Commons License This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.