Serological Diagnosis of Feline Tegumentary Leishmaniasis by Indirect Immunofluorescence (IFI) and Enzyme-Linked Immunosorbent Assay (ELISA) in an Endemic Area in Brazil

Fabiano Borges Figueiredo, Lílian Dias Nascimento, Tassia Cristina Bello de Vasconcelos, Maria de Fátima Madeira, Eliame Mouta Confort, Tânia Maria Valente Pacheco Schubach


Background: American tegumentary leishmaniasis (ATL) is a serious public health problem, and the participation of domestic cats in its epidemiological process has not yet been fully elucidated. Therefore, the standardization of monitoring methodologies of cat populations becomes important for the generation of information on the disease. In Brazil, ATL presents a wide geographical distribution, being Leishmania (Viannia) braziliensis its etiologic agent of ATL in Rio de Janeiro. The main objective of the present study was investigate the presence of specific anti-Leishmania (Viannia) braziliensis Immunoglobulin G (IgG) in 34 cats from an ATL endemic area in this municipality.

Materials, Methods & Results: Sera from three cats from the study area naturally infected with Leishmania (Viannia) braziliensis were used as positive control. Analyses were performed with antigen preparations using indirect immunofluorescence (IFI) and enzyme-linked immunosorbent assay (ELISA). We found prevalence rates of 20.6% (7/34) in the IFI and 14.7% (5/34) in the ELISA. Specificity was 87.1% for the IFI and 93.5% for the ELISA, and both tests presented sensitivity of 100%. Concordance between the used tests was assessed as moderate.

Discussion: By conducting a feline serological survey in an endemic area for ATL we provide information on the involvement of this species in such epidemiological process. In this context, the participation of the cat in cycles of leishmaniases has been highlighted by several authors; infection of the phlebotomine by parasites originated from this vertebrate host can already be demonstrated by xenodiagnosis and several cases of feline infection have been reported in endemic regions of several countries such as Brazil, Spain, France, Portugal, Greece, Iran, Israel and Italy. Therefore, the standardization of diagnostic techniques that support the screening and monitoring of such populations becomes of utmost importance. In fact, the use of the IFI and ELISA serological techniques, showing sensitivity and NPV of 100%, confirm the use of such tools in epidemiological surveys for the detection of anti-L. (V.) braziliensis antibodies in domestic cats, and these methodologies are frequently employed in canine surveys showing good results. The serological titers found in this study for Leishmania (V.) braziliensis are similar to those of studies carried out with cats in endemic areas of visceral leishmaniasis in Spain. This similarity shows that the titers can be high even when L. (V.). braziliensis is the etiological agent. It is worth mentioning that these values are significant when compared with those of canine studies carried out in ATL endemic regions. Ultimately, the sample refers to an ATL endemic region located in Brazil, one of the countries with the highest endemicity levels of cutaneous leishmaniasis in the world and, therefore, an important area for the implementation and evaluation of diagnostic techniques for the generation of data on the disease and its possible hosts. Thus, the present study provides information on the occurrence of ATL by Leishmania (V.) braziliensis in domestic cats; demonstrates that the methodologies employed were favorable to the use of such techniques as tools in epidemiological surveys for the diagnosis of feline tegumentary leishmaniasis; and paves the way for further studies on the subject.


Leishmania (Viannia) braziliensis; domestic cat; serological diagnosis.

Full Text:



Ayllon T., Tesouro M.A., Amusategui I., Villaescusa A., Rodriguez-Franco F. & Sainz A. 2008. Serologic and molecular evaluation of Leishmania infantum in cats from Central Spain. Annals of the New York Academy of Sciences. 1149: 361-364.

Barbosa G.M., Marzochi M.C., Massard C.L., Lima G.P. & Confort E.M. 1999. Epidemiological aspects of canine American tegumentary leishmaniasis in the Municipality of Paraty, State of Rio de Janeiro, Brazil. Cadernos de Saúde Pública. 15: 641-646.

Barnes J.C., Stanley O. & Craig T.M. 1993. Diffuse cutaneous leishmaniasis in a cat. Journal of the American Veterinary Medical Association. 202: 416-418.

Cardoso L., Lopes A.P., Sherry K., Schallig H. & Solano-Gallego L. 2010. Low seroprevalence of Leishmania infantum infection in cats from northern Portugal based on DAT and ELISA. Veterinary Parasitology. 174: 37-42.

Cerqueira A.G.C. & Vasconcelos A. 1922. A leishmaniose nesta capital. Boletim Sanitário. 1: 35.

Costa -Durão J.F., Rebelo E., Peleteiro M.C., Correia J.J. & Simões G. 1994. Primeiro caso de leishmaniose em gato doméstico (Felis catus domesticus) detectado em Portugual (Conselho de Sesimbra). Revista Portuguesa de Ciências Veterinárias. 89: 140-144.

Cupolillo E., Grimaldi Jr. G. & Momen H. 1994. A general classification of new world Leishmania using numeral zymotaxomomy. The American Journal of Tropical Medicine and Hygiene. 50: 296-311.

Da Silva S.M., Rabelo P.F.B., Gontijo N.F., Ribeiro R.R., Melo M.N., Ribeiro V.M. & Michalick M.S.M. 2010. First report of infection of Lutzomyia longipalpis by Leishmania (Leishmania) infantum from a naturally infected cat of Brazil. Veterinary Parasitology. 174: 1-2.

De Souza A.I., Barros E.M.S., Ishikawa E., Ilha I.M.N., Marin G.R.B. & Nunes V.L.B. 2005. Feline leishmaniasis due to Leishmania (Leishmania) amazonensis in Mato Grosso do Sul State, Brazil. Veterinary Parasitology. 128: 41-45.

Diakou A., Papadopoulos E. & Lazarides K. 2009. Specific anti-Leishmania spp. antibodies in stray cats in Greece. The Journal of Feline Medicine and Surgery. 11: 728-730.

Dorbadam S.M., Akhlaghi L., Akhondi B., Hajjaran H., Zarei Z. & Hadighi H. 2014. Evaluation of Leishmania infantum in cat by PCR-RFLP in an endemic region of visceral leishmaniasis in meshkin-shahr, Iran. The Journal Genes, Microbes and Immunity. 2014: 1-7.

Dos Santos G.P., Sanavria A., Marzochi M.C., dos Santos E.G., Silva V.L., Pacheco Rda S., Mouta-Confort E., Espindola C.B., de Souza M.B., Ponte C.S., da Conceicao N.F. & de Andrade M.V. 2005. Prevalence of canine infection from endemic areas of American cutaneous leishmaniasis in Paracambi District, Rio de Janeiro State, between 1992 and 1993. Revista da Sociedade Brasileira de Medicina Tropical. 38: 161-166.

Ferreira W.A., Mayrink W., dos Mares-Guia M.L. & Tavares C.A. 2003. Detection and characterization of Leishmania antigens from an American cutaneous leishmaniasis vaccine for diagnosis of visceral leishmaniasis. Diagnostic Microbiology and Infectious Disease. 45: 35-43.

Figueiredo F.B., Pereira S.A., Gremião I.D.F., Nascimento L.D., Madeira M.F. & Schubach T.M.P. 2008. Leishmaniose tegumentar americana em felino doméstico no município do Rio de Janeiro, Brasil - Relato de caso. Clínica Veterinária. 74: 58-60.

Gontijo B. & de Carvalho M.L. 2003. American cutaneous leishmaniasis. Revista da Sociedade Brasileira de Medicina Tropical. 36: 71-80.

Gontijo C.M., da Silva E.S., de Fuccio M.B., de Sousa M.C., Pacheco R.S., Dias E.S., Andrade Filho J.D., Brazil R.P. & Melo M.N. 2002. Epidemiological studies of an outbreak of cutaneous leishmaniasis in the Rio Jequitinhonha Valley, Minas Gerais, Brazil. Acta Tropica. 81: 143-150.

Hervás J., Chacon M.D.L.F., Sanchez-Isarria M.A., Pellicer S., Carrasco L., Castillo J.A. & Gomez-Villamandos J.C. 1999. Two cases of feline visceral and cutaneous leishmaniosis in Spain. The Journal of Feline Medicine and Surgery. 1: 101-105.

Kirkpatrick C.E., Farrell J.P. & Goldschmidt M.H. 1984. Leishmania chagasi and L. donovani: experimental infections in domestic cats. Experimental Parasitology. 58: 125-131.

Machattie C., Mills E.A. & Chadwick C.R. 1931. Naturally occurring oriental sore of the domestic cat in Iraq. Transactions of the Royal Society of Tropical Medicine and Hygiene. XXV: 103-107.

Maia C. & Campino L. 2011. Can domestic cats be considered reservoir hosts of zoonotic leishmaniasis? Trends in Parasitology. 27 (8): 341-344.

Maia C., Gome J., Cristóvão J., Nunes M., Martins A., Rebêlo E. & Campino L. 2010. Feline Leishmania infection in a canine leishmaniasis endemic region, Portugal. Veterinary Parasitology. 174: 336-340.

Maia C., Nunes M. & Campino L. 2008. Importance of Cats in Zoonotic Leishmaniasis in Portugal. Vector Borne Zoonotic Disease. 8: 555-559.

Maroli M., Pennisi M.G., Di Muccio T., Khoury C., Gradoni L. & Gramiccia M. 2007. Infection of sandflies by a cat naturally infected with Leishmania infantum. Veterinary Parasitology. 145: 357-360.

Martin-Sanchez J., Acedo C., Munoz-Perez M., Pesson B., Marchal O. & Morillas-Marquez F. 2007. Infection by Leishmania infantum in cats: Epidemiological study in Spain. Veterinary Parasitology. 145: 267-273.

Marzochi M.C.A. & Marzochi K.B.F. 1994. Tegumentary and visceral leishmaniasis in Brazil: emerging anthropozoonosis and possibilities for their control. Cadernos de Saúde Pública. 10: 359-375.

Mello G.B. 1940. Verificação da infecção natural do gato (Felix domesticus) por um protozoário do genero Leishmania. Brasil Médico. 54: 180.

Michael S.A., Morsy T.A., El-Seoud S.F. & Saleh M.S. 1982. Leishmaniasis antibodies in stray cats in Ismailiya Governorate, Egypt. Journal of the Egyptian Society of Parasitology. 12: 283-286.

Ministério da Saúde. 2010. Nota técnica conjunta da Secretaria de Vigilância em Saúde do Ministério da Saúde e da Secretaria de Estado da Saúde do Rio Grande do Sul sobre a situação da Leishmaniose Visceral na fronteira do Estado do Rio Grande do Sul com a Argentina. Available in: [Accessed online April 2015].

Nascimento L.D., Passos S.R.L., Mouta-Confort E., Santiago M.A., Alves A.S., Madeira M.F., Schubach A.O. & Marzochi M.C.A. 2009. Comparative study of amplification systems in immunoenzyme assays for the diagnosis of American tegumentary leishmaniasis. Journal of Clinical Laboratory Analysis. 23: 152-156.

Nasereddin A., Salant H. & Abdeen Z. 2008. Feline leishmaniasis in Jerusalem: Serological investigation. Veterinary Parasitology. 158: 364-369.

Ozon C., Marty P., Pratlong F., Breton C., Blein M., Lelievre A. & Haas P. 1998. Disseminated feline leishmaniosis due to Leishmania infantum in Southern France. Veterinary Parasitology. 75: 273-277.

Passos V.M., Lasmar E.B., Gontijo C.M., Fernandes O. & Degrave W. 1996. Natural infection of a domestic cat (Felis domesticus) with Leishmania (Viannia) in the metropolitan region of Belo Horizonte, State of Minas Gerais, Brazil. Memórias do Instituto Oswaldo Cruz. 91: 19-20.

Poli A., Abramo F., Barsotti P., Leva S., Gramiccia M., Ludovisi A. & Mancianti F. 2002. Feline leishmaniosis due to Leishmania infantum in Italy. Veterinary Parasitology. 106: 181-191.

Ribeiro F.C., Schubach A.O., Mouta-Confort E., Schubach T.M., Madeira M.F. & Marzochi M.C.A. 2007. Use of ELISA employing Leishmania (Viannia) braziliensis and Leishmania (Leishmania) chagasi antigens for the detection of IgG and IgG1 and IgG2 subclasses in the diagnosis of American tegumentary leishmaniasis in dogs. Veterinary Parasitology. 148: 200-206.

Rougeron V., Catzeflis F., Hide M., De Meeus T. & Banuls A.L. 2011. First clinical case of cutaneous leishmaniasis due to Leishmania (Viannia) braziliensis in a domestic cat from French Guiana. Veterinary Parasitology. 181: 325-328.

Savani E.S., de Oliveira Camargo M.C., de Carvalho M.R., Zampieri R.A., dos Santos M.G., D’Auria S.R., Shaw J.J. & Floeter-Winter L.M. 2004. The first record in the Americas of an autochthonous case of Leishmania (Leishmania) infantum chagasi in a domestic cat (Felix catus) from Cotia County, São Paulo State, Brazil. Veterinary Parasitology. 120: 229-233.

Schubach T.M., Figueiredo F.B., Pereira S.A., Madeira M.F., Santos I.B., Andrade M.V., Cuzzi T., Marzochi M.C. & Schubach A. 2004. American cutaneous leishmaniasis in two cats from Rio de Janeiro, Brazil: first report of natural infection with Leishmania (Viannia) braziliensis. Royal Society of Tropical Medicine and Hygiene. 98: 165-167.

Sergent E., Sergent E., Lombaard J. & Quilichini M. 1912. La Leishmaniose à Alger. Infection simultanée d´un enfant, d´un chien et d´un chat dans la meme habitation. Bulletin de la Société de pathologie exotique. 5: 93-98.

Serra C.M., Leal C.A., Figueiredo F., Schubach T.M., Duarte R., Uchoa C.M., Silva R.M. & Madeira M.F. 2003. Canine tegumentary leishmaniasis in Morada das Aguias (Serra da Tiririca), Marica, Rio de Janeiro, Brazil. Cadernos de Saúde Pública. 19: 1877-1880.

Shrout P.E. 1998. Measurement reliability and agreement in psychiatry. Statistical Methods in Medical Research. 7: 301-317.

Silva R.C.N., Ramos R.A.N., Pimentel D.S., Oliveira G.M.A., Carvalho G.A., Santana M.A., Faustino M.A.G. & Alves L.C. 2014. Detection of antibodies against Leishmania infantum in cats (Felis catus) from the State of Pernambuco, Brazil. Revista da Sociedade Brasileira de Medicina Tropical. 47(1): 108-109.

Silveira T.G., Teodoro U., Lonardoni M.V., Toledo M.J., Bertolini D.A., Arraes S.M. & Vedovello Filho D. 1996. Serologic survey of dogs in an endemic area of tegumentary leishmaniasis in Parana State, southern Brazil. Cadernos de Saúde Pública. 12: 89-93.

Solano-Gallego L., Rodríguez-Cortés A., Iniesta L., Quintana J., Pastor J., Espada Y., Portús M. & Alberola J. 2007. Cross-sectional serosurvey of feline leishmaniasis in ecoregions around the Northwestern Mediterranean. The American Journal of Tropical Medicine and Hygiene. 76: 676-680.

Torres F.D., Mattos L.S., Brito F.L.C., Figueiredo L.A. & Faustino M.A.G. 2006. Leishmaniose felina: revisão de literatura. Clínica Veterinária (março/abril): 32-40.

World Health Organization (WHO). 2012. Status of endemicity of cutaneous leishmaniasis, worldwide, 2012. Available in: [Accessed online April 2015].

Vita S., Santori D., Aguzzi I., Petrotta E. & Luciani A. 2005. Feline leishmaniasis and ehrlichiosis: serological investigation in Abruzzo region. Veterinary Research Communications. 29: 319-321.


Copyright (c) 2018 Fabiano Borges Figueiredo, Lílian Dias Nascimento, Tassia Cristina Bello de Vasconcelos, Maria de Fátima Madeira, Eliame Mouta Confort, Tânia Maria Valente Pacheco Schubach

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.