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Revista Brasileira de Parasitologia Veterinária

On-line version ISSN 1984-2961

Rev. Bras. Parasitol. Vet. (Online) vol.19 no.4 Jaboticabal Oct./Dec. 2010

https://doi.org/10.1590/S1984-29612010000400013 

RESEARCH NOTE

 

Occurrence of Leishmania (Leishmania) chagasi in a domestic cat (Felis catus) in Andradina, São Paulo, Brazil: case report

 

Ocorrência de Leishmania (Leishmania) chagasi em gato doméstico (Felis catus) em Andradina, São Paulo, Brasil: relato de caso

 

 

Willian Marinho Dourado CoelhoI; Valéria Marçal Felix de LimaII; Alessandro Francisco Talamini do AmaranteIII; Helio LangoniIV; Virgínia Bodelão Richini PereiraIV; Aziz AbdelnourV; Katia Denise Saraiva BrescianiVI

1Departamento de Medicina Veterinária Preventiva, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista – UNESP
IIDepartamento de Clínica, Cirurgia e Reprodução Animal, Faculdade de Odontologia, Universidade Estadual Paulista – UNESP
IIIDepartamento de Parasitologia, Instituto de Biociências, Universidade Estadual Paulista – UNESP
IVDepartamento de Higiene Veterinária e Saúde Pública, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista – UNESP
VFaculdade de Ciências Agrárias e Veterinárias, Fundação Educacional de Andradina – FEA
VIDepartamento de Apoio, Produção e Saúde Animal, Universidade Estadual Paulista – UNESP

Corresponding author

 

 


ABSTRACT

This work describes natural infection by Leishmania in a domestic cat where amastigote forms of the parasite were observed in the popliteal lymph node imprint. Positive and negative serological reactions were observed by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFA), respectively. Polymerase chain reaction (PCR) revealed that the nucleotide sequence of the sample was identical to Leishmania (L.) chagasi. This is the first report of the disease in felines of the city of Andradina, SP, an area considered endemic for canine and human visceral leishmaniasis.

Keywords: Cat, Leishmania chagasi, PCR, serology.


RESUMO

Neste trabalho, é relatada a infecção natural por Leishmania em um gato doméstico no qual, formas amastigotas do parasito foram observadas em imprint de linfonodo poplíteo. Reações sorológicas positivas e negativas foram observadas pelo teste de imunoadsorção enzimática (ELISA) e reação de imunofluorescência indireta (RIFI), respectivamente. A reação em cadeia da polimerase (PCR) revelou que a sequência de nucleotídeos foi idêntica à Leishmania (L.) chagasi. Este é o primeiro relato da doença em felino da cidade de Andradina, Estado de São Paulo, Brasil, área considerada endêmica para leishmaniose visceral canina e humana.

Palavras-chave: Gato, Leishmania chagasi, PCR, sorologia.


 

 

Introduction

The occurrence of visceral leishmaniasis in cats has been described in several countries (MANCIANTI, 2004; MARTÍN-SÁNCHEZ et al., 2007; MAIA et al., 2008; NASEREDDIN et al., 2008), including Brazil (SAVANI et al., 2004; SILVA et al., 2008). Skin lesions have also been reported in cats with leishmaniasis (PENNISI et al., 2004).

Here we describe a case of natural infection of a domestic cat by Leishmania chagasi in the city of Andradina, located in an area endemic for human and canine visceral leishmaniasis. Andradina is in the region of Araçatuba / São Paulo (20.8961º, 51.37944º) at 405 m of altitude.

This study was approved by the Ethics Committee on Animal Experimentation of the Faculty of Dentistry of Araçatuba – UNESP – Protocol No. 2007-003276.

The feline, a male adult mongrel, was given for euthanasia by his owners to the local Center for Animal Control. This animal was from the urban area and had extensive dermatitis with crusts and small ulcers on its face as well as on the palmar and plantar areas of its legs.

Leishmania spp. amastigotes were seen in the feline's lymph node imprints stained with Quick Panoptic kit (Hematocor Biolog®), but not in imprints from bone marrow, spleen, liver, and skin lesions. Analysis of serum sample was negative by IFA (cut-off 1:40) and positive by ELISA performed as described by Lima et al. (2005) with protein A as an antigen, in triplicate. Serology for feline immunodeficiency virus (FIV) and feline leukemia (FeLV) performed with the kit Snap® Combo Plus Test, was negative.

DNA was extracted from lymph nodes and spleen samples using Genomic PrepTM Cells and Tissue DNA GE Healthcare kit. PCR reactions were performed at final volumes of 25 µL containing reaction buffer (10 mM Tris HCl, 50 mM KCl), 1.5 mM MgCl2, 0.2 mM dNTP, 0.2 units Platinum Taq DNA polymerase (Invitrogen), 10ng of extracted DNA, and 10mmol of each of the RV1 and RV2 primers (LACHAUD et al., 2002). RV1 primer sequence (5'-CTTTTCTGGTCCCGCGGGTAG-3') and RV2 primer sequence (5'-CCACCTGGCCTATTTTACACCA -3') produce a 145 bp-conserved region present in all Leishmania species. PCR mixtures were incubated in a thermocycler Mastercycler Gradient (Eppendorf) using the following cycling profile: initial denaturation at 94 ºC for 4 minutes, 40 cycles of 94 ºC for 30 seconds, 59 ºC for 30 seconds, 72 ºC for 30 seconds, and 70 ºC for 10 minutes. The amplified products were analyzed by electrophoresis on 1.5% agarose gel stained with SYBR® safe (Invitrogen®).

The amplified fragments were visualized with an image analyzer (GelDoc-ITTM Imaging System), using the software Vision Works® LS. Next, amplicons were purified with the enzyme ExoSAP-IT (USB) and sequenced on MegaBace 1000 DNA sequencer (GE Healthcare) using the termination kit DYEnanic ET Dye (GE Healthcare). The sequences were compared with those deposited in the NCBI database using blastn (Basic Local Tool for Nucleotide Alignment). Positive fragments of L. chagasi were detected in tissue samples of spleen and bilateral popliteal lymph nodes (Figure 1). Database alignments of the amplified fragments showed 97% identity with the DNA sequences of L. chagasi in the GenBank: Z35276.

 

 

The occurrence of Leishmania in cats has been described by parasitological, molecular (HATAM et al., 2010; TABAR et al., 2008), and serological tests (DIAKOU et al., 2009; SOLANO-GALLEGO et al., 2007) as well as by IFA (MAROLI et al., 2007; AYLLON et al., 2008).

Using parasitological and serological assays, previous studies reported an occurrence of 6.5% (13/200) of animals infected with Leishmania spp., in the municipality of Araçatuba, located at approximately 120 km from Andradina (ROSSI, 2007). In other countries, epidemiological surveys using IFA and PCR revealed a ratio of 30.4% (7/23) in Portugal (MAIA et al., 2008), and 6.7% (7/104) in Israel using ELISA (NASEREDDIN et al., 2008).

Taken together, these studies indicate that cats may play an important role in the epidemiology of leishmaniasis, acting as a possible source of infection (DA SILVA et al., 2010). More detailed studies will be necessary to assess feline's contribution in the maintenance and spread of leishmaniasis in different environmental conditions

According to the data revealed herein it is plausible to think that in an area endemic for canine visceral leishmaniasis, with confirmed reports of human infection, cats may also be affected by the parasite.

 

References

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Corresponding author:
Helio Langoni
Departamento de Higiene Veterinária e Saúde Pública
Faculdade de Medicina Veterinária e Zootecnia
Universidade Estadual Paulista – UNESP
Distrito Rubião Jr., s/n
CEP 18618-970, Botucatu - SP, Brazil
e-mail: hlangoni@fmvz.unesp.br

Received March 16, 2010
Accepted October 6, 2010

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