Characterization of Leishmania species from Central-West Region of Brazil.

Leishmaniasis represents a complex of diseases with a broad clinical spectrum and epidemiological diversity. Involving a variety of agents, reservoirs, and vectors, cutaneous leishmaniasis (CL) presents different transmission patterns, which makes its control especially difficult. Epidemiological characterization with identification of existing Leishmania species in a particular region is essential for understanding this parasitosis epidemiology. Because similar results may be observed in places with overlapping transmission of visceral and cutaneous leishmaniasis, contributing to knowledge about the epidemiological cycle of cutaneous leishmaniasis is of global interest. This study proposes to identify the different species of Leishmania present in human CL cases in the state of Mato Grosso do Sul, Brazil. Slides of imprint tissue were selected from patients diagnosed with CL from 2009 to 2013 at the Laboratório de Parasitologia Clínica, Universidade Federal de Mato Grosso do Sul. The slides were scraped and extracted genetic material was amplified with PCR addressing the Leishmania spp. kDNA region. The reaction product was subjected to RFLP for the specific identification of Leishmania. Forty-three samples (82.7%) amplified with PCR were considered positive. Of these, it was possible to determine the species in 40 samples (76.9%) through analysis of electrophoresis profiles. Leishmania (Viannia) braziliensis, Leishmania (Leishmania) amazonensis, and Leishmania (Leishmania) infantum were identified as the etiologic agents of CL in Mato Grosso do Sul. The results of this study permit a more detailed understanding of CL epidemiology and the geographical distribution of its agents.

Anti-Leishmania antibodies in blood donors from the Midwest region of Brazil.

The aim of the study was to detect asymptomatic infection by Leishmania sp. in blood donors. Serum samples (430) were tested by Immunofluorescent Antibody Test, and an interview with the blood donors was carried out. Antibodies were detected in 15.6% of samples. The variables associated with the infection were: origin of the donor, presence of builds, parks or squares, sick dog in the neighborhood, and neighboring with leishmaniasis. It was observed association between origin of donors and the presence of sick dog. It is important a careful screening of donors, due to the risk of infection through blood transfusion.

Ecological aspects of phlebotomines (Diptera: Psychodidae) in endemic area of visceral leishmaniasis, Campo Grande, State of Mato Grosso do Sul, Brazil.

Aspects of phlebotomine behavior were investigated in the city of Campo Grande, Mato Grosso do Sul state. The insects were captured weekly during December 2003 to November 2005, with Centers for Disease Control light traps at seven different sites including forests and residential areas. In total, 11,024 specimens (7,805 males and 3,219 females) were collected, from which 9,963 (90.38%) were identified as Lutzomyia longipalpis, the proven vector of American visceral leishmaniasis agent. The remaining 9.62% comprised 21 species. L. longipalpis was the most frequent species in all sampled sites, and the first in the ranking of standardized species abundance index. In residential areas this species clearly predominated in the peridomicile (90.96%), in contrast to the intradomicile (9.04%); in animal shelters, it was more numerous in hen houses and prevailed at ground level, inside, and at forest edge around the residences; this aspect is worrying because this insect may remain sheltered in forested environments during the use of insecticides in homes. In the forest environment, other probable or proven vector of cutaneous leishmaniasis agents were also captured such as Lutzomyia whitmani (=Nyssomyia whitmani, sensu Galati), Lutzomyia antunesi (=Nyssomyia antunesi, sensu Galati), and Lutzomyia flaviscutellata (=Bichromomyia flaviscutellata, sensu Galati).

Polymerase chain reaction of peripheral blood as a tool for the diagnosis of visceral leishmaniasis in children.

The diagnosis of visceral leishmaniasis (VL) generally requires the use of invasive tests for the collection of infected tissue (aspirates of bone marrow, spleen, liver or lymph nodes). This difficulty has led to the search for safer and less painful techniques to confirm the occurrence of the disease in children. Polymerase chain reaction (PCR) is a method that is advantageous in that it allows the use of peripheral blood samples for diagnosis. This paper reports the utilisation of PCR on peripheral blood samples to diagnose VL in 45 children in Mato Grosso do Sul, Brazil. This technique is compared with methods carried out using tissue collected by invasive procedures, including direct microscopy, culture and detection of Leishmania DNA by PCR in bone marrow aspirates. The results show that PCR of peripheral blood provides great sensitivity (95.6%) that is similar to that from the PCR of bone marrow aspirates (91.1%) and higher than that achieved with microscopy (80%) or culture (26.7%) methods. PCR of peripheral blood proved to be a suitable tool for the diagnosis of VL in children because it is highly sensitive and safe, with tissue collection being less invasive than in traditional tests.

HIV/AIDS-associated visceral leishmaniasis in patients from an endemic area in Central-west Brazil.

An increase in morbidity associated with visceral leishmaniasis (VL) in human immunodeficiency virus (HIV)/AIDS patients has been described in Africa and the Mediterranean. Despite the high endemicity of VL and HIV-1/AIDS in Brazil, this association has not been thoroughly investigated. Our aim was to evaluate the epidemiologic and clinical characteristics of VL-HIV-1/AIDS cases from Central-west [Mato Grosso do Sul (MS)] Brazil. Medical records of 23 VL-HIV-1/AIDS patients were reviewed. Patients were predominantly adult males (87%) and 34.8% of the patients were intravenous drug users (IVDU). Leishmaniasis was the first opportunistic infection in 60% of the HIV-1 patients. Fever occurred in all patients, although splenomegaly and hepatomegaly were absent in 21.7% of the cases. CD4+ T-cell counts were below 200 cells/mm(3) in 80% of the cases and the counts did not increase after clinical remission despite antiretroviral therapy. The first drug chosen to treat the cases was antimonial, but the therapeutic regimen was altered to amphotericin B in 12 of 17 cases due to side effects. Relapses were reported in 56.5% of the patients. IVDU may constitute an important risk factor for the transmission of both diseases in MS. VL-HIV-1/AIDS patients in MS share similar clinical characteristics as those from other endemic regions worldwide. Thus, these findings are critical for improving the surveillance of VL-HIV/AIDS patients.

Presence of anti-Leishmania antibodies in candidates for kidney transplantation.

OBJECTIVES: Visceral leishmaniasis (VL) is a progressive disease that, left untreated, is typically fatal. The purpose of this investigation was to detect Leishmania sp. infection in hemodialysis patients who had received multiple blood transfusions at a private clinic in Campo Grande, Mato Grosso do Sul state, Midwest Brazil. METHODS: Fifty randomly selected volunteers were interviewed for collection of demographic, socioeconomic, and epidemiological data. Indirect immunofluorescence (titers positive when ≥1:40) and rK39 immunochromatographic tests were employed for serological investigation. RESULTS: Males predominated (60%). Age ranged from 20 to 77 years. Most subjects reported being on hemodialysis for at least one year (94%) and 84% were candidates for kidney transplantation, 67% of whom were on the waiting list. Leishmania sp. infection was detected in 32%. Contact with infected dogs was the only variable associated with infection. CONCLUSIONS: Under immunocompromised conditions, VL is opportunistic and potentially fatal. Despite existing risks, screening for VL is not performed in asymptomatic donors and recipients. The detection of anti-Leishmania antibodies in these patients reinforces the need for infection screening before immunosuppressive treatment is initiated to reduce not only the risks of VL development and severity, but also mortality rates in cases of reactivation of latent infection.

Severe adverse reactions to meglumine antimoniate in the treatment of visceral leishmaniasis: a report of 13 cases in the southwestern region of Brazil.

Antimony-based medications continue to be the chosen drug for visceral leishmaniasis treatment in most countries. Pentavalent antimony compounds are highly effective but frequently have adverse reactions. Although toxic effects are almost always reversible, some of them can be severe. Clinical and laboratory data of 13 patients who developed severe adverse reactions to meglumine antimoniate in a teaching hospital in southwestern Brazil in 2004-2005 were analysed. Most patients were adults (10/13), mainly at the age of 50 or older (4/13). The main severe adverse reactions were renal failure (eight episodes), pancreatitis (six episodes) and hepatic failure/hepatitis (five episodes). Six patients died in the period; all presented acute renal failure and four presented hepatic failure. Meglumine antimoniate can cause severe reactions, which can lead to death if not promptly identified. Further studies are warrented on the effect of less toxic drugs.

Disney Trap Attractiveness for the Cortelezzii Series (Diptera: Psychodidae, Phlebotominae) in a Fishing Tourism Area, Bonito Municipality, Mato Grosso do Sul State, Brazil.

Sand fly fauna is frequently sampled using automatic light or Shannon traps, yet few studies have been devoted to investigating the effectiveness of Disney traps baited with live animals as an attractant. This study sought to identify the phlebotomine fauna attracted to Disney traps having hamsters (Mesocricetus auratus) as bait. A hamster-baited Disney trap was installed in a gallery forest located in Águas do Miranda District, Bonito Municipality, Mato Grosso do Sul state, Brazil, from November 2011 to October 2012. A total of 717 phlebotomines were collected, in which male (251) and female (463) specimens of the Cortelezzii series (Diptera: Psychodidae) predominated (99%). Males were attracted from January to July, whereas the females were predominantly attracted from January to May. No significant correlation was observed between the monthly climatic averages of temperature, rains and humidity, and the numbers of insects collected. Although these findings showed the attractiveness of both sexes of the species of the Cortelezzii series to golden hamsters, further studies are needed to investigate the blood meal preferences of these females to other rodents. As rodents have been reported as reservoirs of Leishmania spp. (Kinetoplastida: Trypanosomatidae), research using live-baited traps can reveal feeding preferences of sand flies and the importance of various rodent species in the zoonotic cycle.

Leishmania (Viannia) braziliensis type 2 as probable etiological agent of canine cutaneous leishmaniasis in Brazilian Amazon.

Canine cutaneous leishmaniasis (CCL) is a zoonosis of public health interest, and in the Americas, Leishmania (Viannia) braziliensis has been identified as the main etiological agent. The present study sought to investigate Leishmania spp. infection in domestic dogs from a rural area of the Xapuri municipality, Acre state, Brazilian Amazonia. For this purpose, visits were carried out to domiciles where the human cases of American cutaneous leishmaniasis (ACL) occurred, followed by the clinical evaluation of the animals in search of clinical signs suggestive of CCL. Blood samples were collected from 40 dogs, 13 of which had lesions suggestive of CCL, and biopsies of these lesions were performed. The methods used were Neal, Novy, and Nicolle's (NNN) medium cultures and direct parasitological examination. Further, to detect and characterize Leishmania DNA some molecular techniques were performed such as conventional polymerase chain reaction (PCR) and sequencing targeting SSU rDNA and ITS1, restriction fragment length polymorphism (RFLP) and high resolution melting (HRM) analysis targeting hsp70. The investigation revealed that the results obtained from the parasitological methods were negative. In PCR by ITS1 and network topology sequences, six strains from dogs, isolated from the Peruvian Andes, appeared identical to Leishmania (Viannia) braziliensis type 2 (99-100%). By other molecular methods these samples turned out to be positive to Leishmania (Viannia) sp.. The diagnosis of Leishmania in domestic dogs from Acre state showed a high proportion of infected animals, and the occurrence of L. braziliensis type 2 in Brazil for the first time. This new report suggests that L. braziliensis type 2 is both trans- and cis-Andean. However, more studies are needed regarding the clinical and diagnostic aspects of this species of Leishmania.

Seasonal variation of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) in endemic area of visceral leishmaniasis, Campo Grande, state of Mato Grosso do Sul, Brazil.

The seasonal distribution of Lutzomyia longipalpis was studied in two forested and five domiciliary areas of the urban area of Campo Grande; MS, from December 2003 to November 2005. Weekly captures were carried out with CDC light traps positioned on ground and in the canopy inside a residual forest and on the edge (ground) of a woodland and in at least one of the following ecotopes in peridomiciles-a cultivated area, a chicken coop, a pigsty, a kennel, a goat and sheep shelter and an intradomicile. A total of 9519 sand flies were collected, 2666 during the first year and 6853 during the second. L. longipalpis was found throughout the 2-year period, presenting smaller peaks at intervals of 2-3 months and two greater peaks, the first in February and the second in April 2005, soon after periods of heavy rain. Only In one of the woodlands was a significant negative correlation (p<0.05) between the number of insects and temperature during the first year and the climatic factors (temperature, RHA and rain) was observed. In the domiciliary areas in four domiciles some positive correlations (p< or =0.05) occurred in relation to one or more climatic factors; however, the species shows a clear tendency to greater frequency (72%) in the rainy season than in the dry (28%). Thus, we recommend an intensification of the VL control measures applied in Campo Grande, MS, during the rainy season with a view to reducing the risk of the transmission of the disease.

Enteroparasites in Riverside Settlements in the Pantanal Wetlands Ecosystem.

BACKGROUND: Intestinal parasites are a major source of health problems in developing countries, where socioeconomic, cultural, and environmental conditions contribute in maintaining the biological cycles of various parasites and facilitating their spread. The objective of this study, conducted in Corumbá, Mato Grosso do Sul state, Brazil, was to investigate the occurrence of intestinal parasites in riverside communities in the South Pantanal wetlands and conduct educational interventions focused on health and environmental preservation. METHOD: In total, 196 stool samples were tested for parasites using the merthiolate-iodine-formaldehyde concentration (MIFC) technique and spontaneous sedimentation and educational activities were carried out. RESULTS: Enteroparasite prevalence was 72% (65.6-78.2%; 95% CI). Of the 141 positive cases, monoparasitism was found in 34.7%, biparasitism in 23%, and polyparasitism in 14.3%. Entamoeba coli was the most frequent protozoan (70.2%). Among helminths, hookworms were the most prevalent. Enteroparasitosis prevalence did not differ for sex or place of abode but proved higher in individuals older than 10 years. CONCLUSION: The high positivity rate for enteroparasites found for the communities stems from lack of sanitation and poor personal and environmental hygiene habits, indicating that effective health policies and educational interventions are needed to reduce the current risk levels.

Prevalence of intestinal parasites among inmates in Midwest Brazil.

BACKGROUND: Intestinal parasitic infections constitute a public health issue in developing countries, with prevalence rates as high as 90%, a figure set to escalate as the socioeconomic status of affected populations deteriorates. Investigating the occurrence of these infections among inmates is critical, since this group is more vulnerable to the spread of a number of infectious illnesses. METHODS: This cross-sectional, analytical, quantitative study was conducted in July 2015 at prison facilities located in Midwest Brazil to estimate the prevalence of parasitic infection among inmates. For detection of parasites, 510 stool samples were examined by ether centrifugation and spontaneous sedimentation. RESULTS: Eight parasitic species were detected, with an overall prevalence of 20.2% (103/510). Giardia lamblia and Entamoeba histolytica/dispar were the most frequent pathogenic parasites. Endolimax nana was the predominant non-pathogenic species. Nearly half of the subjects (53/103; 51.4%) were positive for mixed infection. Logistic regression revealed that inmates held in closed conditions were more likely to contract parasitic infections than those held in a semi-open regime (OR = 1.97; 95% CI = 1.19-3.25; p = 0.0085). A higher prevalence of parasitic infections was observed among individuals who had received no prophylactic antiparasitic treatment in previous years (OR = 10.2; 95% CI = 5.86-17.66; p < 0.001). The other factors investigated had no direct association with the presence of intestinal parasites. CONCLUSION: Infections caused by directly transmissible parasites were detected. Without adequate treatment and prophylactic guidance, inmates tend to remain indefinitely infected with intestinal parasites, whether while serving time in prison or after release.

Leishmania (V.) braziliensis infecting bats from Pantanal wetland, Brazil: First records for Platyrrhinus lineatus and Artibeus planirostris.

In the New World genus Leishmania parasites are etiological agents of neglected zoonoses known as leishmaniasis. Its epidemiology is very complex due to the participation of several species of sand fly vectors and mammalian hosts, and man is an accidental host. Control is very difficult because of the different epidemiological patterns of transmission observed. Studies about Leishmania spp. infection in bats are so scarce, which represents a large gap in knowledge about the role of these animals in the transmission cycle of these pathogens, especially when considering that Chiroptera is one of the most abundant and diverse orders among mammals. Leishmaniasis in Mato Grosso do Sul, Brazil are remarkably frequent, probably due to the abundance of its regional mastofauna. The recent record of L. braziliensis in bats from this state indicates the need to clarify the role of these mammals in the transmission cycle. In this study we evaluated the presence of Leishmania parasites in the skin of different species of bats, using PCR directed to Leishmania spp. kDNA for screening followed by PCR/RFLP analysis of the hsp70 gene for the identification of parasite species. Leishmania species identification was confirmed by PCR directed to the G6PD gene of L. braziliensis, followed by sequencing of the PCR product. Samples from 47 bats were processed, of which in three specimens (6.38%) was detected the presence of Leishmania sp. kDNA. PCR/RFLP and sequencing identified the species involved in the infection as L. braziliensis in all of them. This is the first report of Leishmania braziliensis in bats from Pantanal ecosystem and the first record of this species in Platyrrhinus lineatus and Artibeus planirostris, bats with a wide distribution in South America. These results reinforce the need to deepen the knowledge about the possibility of bats act as reservoirs of Leishmania spp. especially considering their ability of dispersion and occupation of anthropic environments.

[Emergent outbreak of visceral leishmaniasis in Mato Grosso do Sul State]. / Foco emergente de leishmaniose visceral em Mato Grosso do Sul.

This study, realized from October 2000 to January 2003 describes the clinical epidemiological characteristics of visceral leishmaniasis in Três Lagoas, Mato Grosso do Sul State, Brazil. A total of 149 cases were confirmed, with a predominance of the male gender (71.1%). The principal age group was aged 0 to 4 years old (42%). The clinical picture included fever (97.3%), esplenomegaly (85.9%) and anemia (75.8%). Associated infections were seen in 32 patients (21.5%), pneumonia being most common. Changes registered in the laboratory included a median hemoglobin level of 8mg/dl and 3,100 leucocytes/mm(3). Bone marrow smears were positive in 90.6% of patients. Of the 97.9% patients treated, 78.2% used pentavalent antimony. Mortalities occurred in 8% of cases, half of these with associated infection. Taken together, these data suggest changes in the physiographical occurrence of visceral leishmaniasis in this locality, with expansion and urbanization of the disease, requiring greater attention for early diagnosis and treatment.

Sandflies in an urban area of transmission of visceral leishmaniasis in midwest Brazil.

The phlebotomine fauna of Campo Grande city, capital of Mato Grosso do Sul state in Brazil, an endemic area for visceral leishmaniasis, has been thoroughly investigated, but all the insect collections were undertaken with automatic light traps. The present study sought to investigate the fauna in this city using Shannon and Disney traps, having human beings and hamsters, respectively, as bait. Both types of traps were installed in forest fragment and peridomiciliary areas in the period from 2007 to 2009. The phlebotomine females were analyzed by PCR for Leishmania identification. Lutzomyia longipalpis was the only species collected in the peridomiciles and rendered a total of 574 specimens with a 5.2:1 male:female ratio. A total of eight species were attracted to the two traps (one of each type) installed in the forest fragment, including: Bichromomyia flaviscutellata, Evandromyia bourrouli, Evandromyia lenti, Lutzomyia longipalpis, Nyssomyia whitmani, Pintomyia christenseni, Psathyromyia bigeniculata, and Sciopemyia sordellii. A total of 143 specimens were collected, Bi. flaviscutellata accounting for 81% and Lu. longipalpis for 1.4% of them. In one female of Lu. longipalpis collected in a Disney trap installed in a peridomicile, Leishmania (Leishmania) infantum DNA was found, thus strengthening the hypothesis that the transmission of leishmaniasis is in fact occurring in the anthropic environment.

Antissaliva Antibodies of Lutzomyia Longipalpis in area of Visceral Leishmaniasis.

The aim of the present study was to assess the presence of antissaliva antibodies of Lutzomyia longipalpis in human hosts living in area of visceral leishmaniasis, located in the Center-West region of Brazil. The presence of antissaliva antibodies of L. longipalpis exhibited a strong correlation with the protection and development of antibodies against Leishmania sp. Of the 492 children studied, elevated antissaliva antibodies of L. longipalpis were detected in 38.4% of the participants. There was a higher percentage of positivity (64.7%) among children who exhibited anti-Leishmania sp. antibodies and among those who were positive in the delayed hypersensitivity test (34.8%).

Late diagnosis: a factor associated with death from visceral leishmaniasis in elderly patients.

INTRODUCTION: Visceral leishmaniasis (VL) is among the seven global endemic diseases assigned a high priority by the World Health Organization. In Latin America, most cases occur in Brazil. Despite the availability of intensive treatment resources and protocols for specific treatment, lethality rates for VL have increased in several regions in the country over the past 10 years, particularly in patients under one and over 50 years of age. As the growth of the elderly population accelerates in Brazil, VL poses a greater challenge to public health. Given the scarcity of studies addressing the disease in this age group, the purpose of this study was to identify factors associated with VL lethality among the elderly. METHODS/KEY FINDINGS: This analytical, cross-sectional epidemiological study comprised 80 elderly patients who sought treatment at the teaching hospital of the Universidade Federal de Mato Grosso do Sul, Campo Grande, MS, Brazil, in the period 2000-2013.Clinical, laboratory and treatment variables were investigated from records of elderly patients with VL diagnosis confirmed by at least one laboratory test positive (culture for parasite or direct parasitological examination; reactive immunofluorescence; immunochromatographic test with recombinant antigens) or patients without laboratory confirmation who lived in endemic areas and responded favorably to therapeutic trial, as defined by the Brazilian Ministry of Health. Of the 80 patients included, 78 tested positive to at least one exam; in two cases, diagnosis was based on clinical and epidemiological criteria. The lethality rate was 20%. Multivariate analysis revealed an association between death and time elapsed from symptom onset.

First record of Leishmania braziliensis presence detected in bats, Mato Grosso do Sul, southwest Brazil.

Leishmaniasis, a zoonotic disease caused by parasites of the genus Leishmania, has expanded beyond its natural range and is becoming increasingly urban. Using PCR and PCR-RFLP, we detected Leishmania (Viannia) braziliensis in two bats (Chiroptera) in Mato Grosso do Sul, Brazil, an endemic area. This is the first record of L. (V.) braziliensis in bats. It is also the first record of any Leishmania sp. in bats in the state. The animals testing positive were found in both a rural site and an urban site. These results indicate the need for further research into the viability of Leishmania in bats and could potentially have implications for public health in Mato Grosso do Sul, given the large populations of urban bats, their mobility, and their ability to roost at close proximity to humans within residences and other buildings.

Canine visceral leishmaniosis in an area of fishing tourism, Bonito Municipality, Mato Grosso do Sul, Central-West Brazil / Leishmaniose visceral canina em área de turismo pesqueiro, município de Bonito, Mato Grosso do Sul, Centro-Oeste do Brasil

O estudo teve como objetivo verificar a infecção por Leishmania em uma população de cães (n=92 animais) de uma área de turismo de pesca no município de Bonito e avaliar comparativamente os métodos sorológicos utilizados, ensaio imunoenzimático (ELISA), reação de imunofluorescência indireta (RIFI) e teste rápido de plataforma dupla (DPP®). Testes parasitológicos também foram realizados, como o isolamento do parasita em meio de cultura Neal, Novy, Nicolle, técnica de Woo, esfregaços em lâminas coradas com Giemsa e identificação específica pela reação em cadeia da polimerase (PCR). A RIFI revelou sorologia positiva de 56/92, ELISA 8/92 e DPP® 41/92. Quanto à concordância entre os testes sorológicos pelo índice Kappa, houve um ligeiro acordo entre ELISA/RIFI e ELISA/DPP®, e uma concordância razoável entre RIFI/DPP®. A cultura NNN foi positiva em três cães, e identificada por PCR como Leishmania infantum chagasi. Nenhuma amostra foi positiva pela técnica de Woo. Nossos resultados mostraram baixas concordâncias entre os testes sorológicos recomendados pelo Ministério da Saúde, sendo necessário associar a outras técnicas de diagnóstico, como testes parasitológicos e PCR, para aumentar a sensibilidade ao diagnóstico de leishmaniose visceral canina, principalmente, com relação aos cães assintomáticos em áreas endêmicas.

Canine visceral leishmaniosis in an area of fishing tourism, Bonito Municipality, Mato Grosso do Sul, Central-West Brazil / Leishmaniose visceral canina em área de turismo pesqueiro, município de Bonito, Mato Grosso do Sul, Centro-Oeste do Brasil

ABSTRACT: The study aimed to investigate Leishmania infection in a population of dogs (92 animals) in a fishing area of Bonito Municipality, and evaluate comparatively the serological methods used, immunoenzymatic assay (ELISA), indirect immunofluorescence antibody test (IFAT), and rapid Dual Path-Platform test (DPP®). Blood and aspirate of bone marrow samples were used and parasitological investigation was also performed, such as parasite isolation in Neal, Novy, Nicolle (NNN) medium culture, Woo technique, Giemsa stained smears and specific identification by polymerase chain reaction (PCR). IFAT revealed 56/92 seropositive, the ELISA 8/92 and the DPP®41/92. Regarding the agreement between the serological tests by the Kappa index, there was a slight agreement between ELISA/IFAT and ELISA/DPP®, and fair agreement in IFAT/DPP®. The NNN culture was positive in three out of 21 dogs, and identified by PCR as Leishmania infantum chagasi. No samples were positive by the Woo technique. Our results showed low agreements between the serological tests recommended by the Brazilian Ministry of Heath, and it is necessary to associate other diagnostic techniques, such as parasitological tests and PCR, to increase the sensitivity for canine visceral leishmaniosis diagnosis, mainly regarding asymptomatic dogs in endemic areas.

RESUMO: O estudo teve como objetivo verificar a infecção por Leishmania em uma população de cães (n=92 animais) de uma área de turismo de pesca no município de Bonito e avaliar comparativamente os métodos sorológicos utilizados, ensaio imunoenzimático (ELISA), reação de imunofluorescência indireta (RIFI) e teste rápido de plataforma dupla (DPP®). Testes parasitológicos também foram realizados, como o isolamento do parasita em meio de cultura Neal, Novy, Nicolle, técnica de Woo, esfregaços em lâminas coradas com Giemsa e identificação específica pela reação em cadeia da polimerase (PCR). A RIFI revelou sorologia positiva de 56/92, ELISA 8/92 e DPP®41/92. Quanto à concordância entre os testes sorológicos pelo índice Kappa, houve um ligeiro acordo entre ELISA/RIFI e ELISA/DPP®, e uma concordância razoável entre RIFI/DPP®. A cultura NNN foi positiva em três cães, e identificada por PCR como Leishmania infantum chagasi. Nenhuma amostra foi positiva pela técnica de Woo. Nossos resultados mostraram baixas concordâncias entre os testes sorológicos recomendados pelo Ministério da Saúde, sendo necessário associar a outras técnicas de diagnóstico, como testes parasitológicos e PCR, para aumentar a sensibilidade ao diagnóstico de leishmaniose visceral canina, principalmente, com relação aos cães assintomáticos em áreas endêmicas.