Canine visceral leishmaniasis in area with recent Leishmania transmission: prevalence, diagnosis, and molecular identification of the infecting species

Abstract INTRODUCTION: Canine visceral leishmaniasis (CVL) is an endemic disease in Brazil, and integrated control actions have been adopted by the Brazilian Ministry of Health to control its spread. However, the transmission profile is unknown in areas with recent CVL cases, including Itaúna, located in the Brazilian state of Minas Gerais, where the present study was carried out. METHODS: A total of 2,302 dogs from 12 neighborhoods were serologically tested for canine VL using the current diagnostic protocol adopted by the Brazilian Ministry of Health. Test positivity rate (TPR) and CVL prevalence were determined for each neighborhood. The presence of Leishmania was assessed in 60 seropositive dogs which had been recommended for euthanasia. Twenty-two of them (37%) were asymptomatic, and 38 (63%) were symptomatic for CVL. Parasitological (myeloculture and smear/imprint) and molecular (PCR) methods were employed for Leishmania detection in bone marrow, spleen, mesenteric lymph nodes, and ear skin. The infecting Leishmania species was identified by DNA sequencing. RESULTS: CVL prevalence (per 1,000 dogs) varied from 0.0-166.67, depending on the neighborhood, with a mean of 68.96 (SD 51.38). Leishmania DNA was detected in at least one tissue from all seropositive dogs, with comparable TPR among tissues. Leishmania parasites were identified in most (54/60) seropositive dogs, and the infecting parasite was identified as Leishmania infantum in all of these. CONCLUSIONS: Prevalence of CVL is a contributor to the spread of visceral leishmaniasis in Itaúna.

Evaluation of parasitological examination, kDNA polymerase chain reaction and rK39-based immunochromatography for the diagnosis of visceral leishmaniasis in seropositive dogs from the screening-culling program in Brazil

Introduction Dogs play a primary role in the zoonotic cycle of visceral leishmaniasis (VL). Therefore, the accurate diagnosis of infected dogs, primarily asymptomatic dogs, is crucial to the efficiency of VL control programs. Methods We investigated the agreement of four diagnostic tests for canine visceral leishmaniasis (CVL): parasite detection, either after myeloculture or by direct microscopic examination of tissue imprints; kinetoplast-deoxyribonucleic acid-polymerase chain reaction (kDNA-PCR); and an immunochromatographic test (ICT). An enzyme-linked immunosorbent assay (ELISA) and an indirect immunofluorescence test (IFAT), both of which were adopted as part of the screening-culling program in Brazil, were used as reference tests. Our sample set consisted of 44 seropositive dogs, 25 of which were clinically asymptomatic and 19 were symptomatic for CVL according to ELISA-IFAT. Results The highest and lowest test co-positivities were observed for ICT (77.3%) and myeloculture (58.1%), respectively. When analyzed together, the overall percentage of co-positive tests was significantly higher for the symptomatic group compared to the asymptomatic group. However, only ICT was significantly different based on the results of a separate analysis per test for each group of dogs. The majority (93.8%) of animals exhibited at least one positive test result, with an average of 2.66 positive tests per dog. Half of the symptomatic dogs tested positive for all four tests administered. Conclusions The variability between test results reinforces the need for more efficient and reliable methods to accurately diagnose canine VL, particularly in asymptomatic animals. .

Assessment of sand fly (Diptera, Psychodidae) control using cypermethrin in an endemic area for visceral leishmaniasis, Montes Claros, Minas Gerais State, Brazil / Avaliação do controle de flebotomíneos (Diptera, Psychodidae) usando cipermetrina em área endêmica para leishmaniose visceral, Montes Claros, Minas Gerais, Brasil

Montes Claros in Minas Gerais State, Brazil, was considered an intense transmission area for visceral leishmaniasis. This study evaluated sand fly fauna after insecticide application. Captures were performed in 10 districts from September 2005 to August 2006 with CDC light traps inside and outside each residence. Cypermethrin was sprayed in two cycles during November/2005 and May/2006. The 636 specimens collected, belonging to 10 species, were predominantly Lutzomyia longipalpis (79 percent), and most frequently males (70 percent). The highest percentage of specimens were captured in areas surrounding domiciles (85.8 percent). The main species were observed to be sensitive to treatment with the insecticide. The results showed a reduction in the number of sand flies collected after use of cypermethrin in homes and annexes, and with residual effect lasting from two to four months.

Montes Claros foi considerada área de transmissão intensa para leishmaniose visceral no Estado de Minas Gerais, Brasil. Este trabalho avaliou a fauna de flebotomíneos após a aplicação do inseticida. Entre setembro de 2005 e agosto de 2006, foram realizadas capturas com 20 armadilhas luminosas CDC em 10 bairros do município, no intra e no peridomicílio de cada residência. Dois ciclos de borrifação com cipermetrina foram realizados nos meses de novembro/2005 e maio/2006. Coletou-se 636 exemplares pertencentes a 10 espécies, com predominância de Lutzomyia longipalpis (79 por cento). Machos foram coletados com maior frequência (70 por cento). O peridomicílio apresentou a maior porcentagem dos espécimens capturados (85,8 por cento). Observou-se que as principais espécies foram sensíveis ao tratamento com o inseticida. Os resultados mostraram uma redução do número de flebotomíneos coletados devido ao uso de cipermetrina nos domicílios e seus anexos, mas com efeito residual atuante entre dois e quatro meses.

Infecção natural de Lutzomyia (Lutzomyia) longipalpis (Diptera: Psychodidae) por Leishmania infantum chagasi em flebotomíneos capturados no município de Janaúba, Estado de Minas Gerais, Brasil / Natural infection with Leishmania infantum chagasi in Lutzomyia (Lutzomyia) longipalpis (Diptera: Psychodidae) sandflies captured in the municipality of Janaúba, State of Minas Gerais, Brazil

INTRODUÇÃO: A leishmaniose visceral tem sido notificada em quase todos os estados do Brasil, e principalmente no norte de Minas Gerais, onde a doença é endêmica. Este estudo visou detectar a infecção natural de Lutzomyia longipalpis e identificar através da técnica de PCR/RFLP a espécie de Leishmania encontrada nos flebotomíneos do município de Janaúba. MÉTODOS: Utilizando-se armadilhas luminosas, foram capturadas 1.550 fêmeas de L. longipalpis, que agrupadas em pool de 10 exemplares foram submetidas à extração e amplificação de DNA, através das técnicas de PCR genérico e cacofonia. RESULTADOS: Dos 155 pools, seis apresentaram-se positivos para Leishmania sp., sendo a taxa de infecção do município de 3,9 por cento. Através da PCR/RFLP determinou-se que o padrão de digestão das amostras positivas foi semelhante ao da cepa referência Leishmania chagasi (MHOM/BR/74/PP75). CONCLUSÕES: A detecção de infecção natural associada a estudos sobre a epidemiologia da LV sugere que L. longipalpis esteja envolvida na transmissão de L. infantum chagasi em Janaúba, principalmente nas áreas de intensa transmissão de LV.

INTRODUCTION: Visceral leishmaniasis has been notified in nearly all states of Brazil, and particularly in the north of Minas Gerais, where the disease is endemic. The aim of this study was to detect natural infection of Lutzomyia longipalpis and, through the PCR/RFLP technique, identify Leishmania species found in sandflies in the municipality of Janaúba. METHODS: Using light traps, 1,550 females of L. longipalpis were caught and grouped into pools of 10 specimens to be subjected to DNA extraction and amplification, by means of generic PCR and cacophony. RESULTS: Out of the 155 pools, six were positive for Leishmania sp., and thus the infection rate in the municipality was 3.9 percent. Through PCR/RFLP, the digestion pattern among the positive samples was found to be similar to that of the reference strain of Leishmania chagasi (MHOM/BR/74/PP75). CONCLUSIONS: The detection of natural infection associated with studies on the epidemiology of visceral leishmaniasis suggests that L. longipalpis is involved in transmission of L. infantum chagasi in Janaúba, particularly in areas of intense transmission of visceral leishmaniasis.

Phlebotominae distribution in Janaúba, an area of transmission for visceral leishmaniasis in Brazil

In Brazil, visceral leishmaniasis (VL) is caused by Leishmania chagasi parasites that are transmitted to man through the bites of infected females of Lutzomyia longipalpis sand flies. In order to evaluate transmission risk and to clarify the epidemiology of this tropical disease, studies focused on the vector and favorable environmental conditions are of fundamental importance. In this work, we surveyed the phlebotomine sand fly fauna in Janaúba, a Brazilian municipality that is endemic for VL. During a two-year period, entomological captures were performed monthly in 15 districts with high, moderate and low profiles of VL transmission. A total of 14,591 phlebotomine sand flies were captured (92 percent L. longipalpis), with a predominance of males. Most specimens were captured in the peri-domicile setting, although the number of specimens captured in the intra-domicile setting emphasises the anthropophilic behaviour of this insect. The population density of L. longipalpis was modulated by climate variations, particularly with clear increases immediately after the rainy season. However, the pattern of distribution did not coincide with the occurrence of human or canine cases of VL. This suggests that the eco-epidemiology of VL is particular to each area of transmission and must be taken into account during the design of public health control actions.

Flebotomíneos (Diptera: Psychodidae) de um foco de leishmaniose tegumentar no Estado de Minas Gerais / Sandflies (Diptera: Psychodidae) in an outbreak of cutaneous leishmaniasis in the State of Minas Gerais

Na localidade do Brejo do Mutambal, situado no município de Varzelândia (MG), área endêmica para leishmaniose tegumentar, foram realizadas capturas mensais sistemáticas utilizando-se armadilhas luminosas do tipo CDC durante o período de janeiro a dezembro de 2000. Foram capturadas 19 espécies de flebotomíneos, totalizando 6.756 exemplares. Lutzomyia intermedia (5,1 por cento), L. migonei (0,4 por cento) e L. whitmani (0,1 por cento), relacionadas com a transmissão de leishmaniose tegumentar, foram capturadas em número reduzido. Lutzomyia longipalpis foi a espécie predominante (34,8 por cento), sugerindo também um risco de transmissão da leishmaniose visceral. A proporção de insetos capturados no peridomícilio foi de 91,7 por cento enquanto no intradomicílio foi de 8,3 por cento. A interferência de fatores climáticos (temperatura, umidade relativa do ar e pluviosidade) sobre a dinâmica populacional de flebotomíneos foi avaliada.

Between January and December 2000, systematic monthly capture of phlebotomine sandflies was undertaken by means of light traps of CDC type in Brejo do Mutambal, an endemic area for American cutaneous leishmaniasis located in the municipality of Varzelândia, State of Minas Gerais. Nineteen phlebotomine species were captured, totaling 6,756 specimens. The species correlated with ACL transmission were captured in low numbers, as follows: Lutzomyia intermedia (5.1 percent), L. migonei (0.4 percent) and L. whitmani (0.1 percent). Lutzomyia longipalpis was the predominant species captured (34.8 percent), thus also suggesting a risk of visceral leishmaniasis transmission. The proportion of sandflies captured in areas surrounding homes was 91.7 percent while 8.3 percent were captured inside homes. The interference of climatic factors (temperature, relative air humidity and rainfall) on phlebotomine population dynamics was evaluated.

Aspectos da ecologia e do comportamento de flebotomíneos em área endêmica de leishmaniose visceral, Minas Gerais / Aspects of the ecology and behaviour of phlebotomines in endemic area for visceral leishmaniasis in State of Minas Gerais

O comportamento e hábitos alimentares de algumas espécies da flebotomíneos têm sido útil na compreensão da epidemiologia das leishmanioses. No município de Porteirinha (MG), foram realizadas capturas mensais sistematizadas utilizando-se 28 armadilhas luminosas tipo CDC, durante o período de janeiro a dezembro de 2002. Foram capturadas 14 espécies de flebotomíneos, totalizando 1.408 exemplares. De acordo com o ambiente, os resultados obtidos mostraram que o peridomicílio apresentou a maior (53,3 por cento) porcentagem dos espécimens encontrados na região, embora parte (46,7 por cento) da fauna também tenha sido encontrada no intradomicílio. O repasto sanguíneo de 38 fêmeas de Lutzomyia longipalpis, provenientes do campo, foi identificado através da reação de precipitina. Os resultados indicam que Lutzomyia longipalpis foi a espécie predominante (65,1 por cento), mostrando-se oportunista, podendo sugar uma ampla variedade de vertebrados.

HP: um modelo aprimorado de armadilha luminosa de sucção para a captura de pequenos insetos / HP: an improved model of sucction light trap for the capture of small insects

Descreve-se um novo tipo de armadilha, resultado do aprimoramento de um modelo muito utilizado, conhecido como CDC. A nova armadilha, denominada HP, apresenta como vantagens alta eficiência, facilidade no manuseio e transporte, além de ser confeccionada quase que exclusivamente com material nacional, o que reduz consideravelmente seu custo final.

Histopathology and immunocytochemical study of type 3 e and type 4 complement receptors in the liver and spleen of dogs naturally and experimentally infected with Leishmania (Leishmania) chagasi

Os objetivos deste trabalho visaram uma analise comparativa das alteracoes histopatologicas e da expressao de CR3 e CR4 no figado e baco de caes natural e experimentalmente infectados com L. chagasi. As lesoes histopatologicas fundamentais observadas principalmente nos caes naturalmente infectados foram: os granulomas epitelioides hepaticos, a hiperplasia e a hipertrofia das celulas de Kupffer, dos foliculos de Malpighi e das celulas monucleadas da polpa vermelha do baco. Os cortes de figado e baco corados pela tecnica de imunocitoquimica mostraram a presenca dos antigenos CD11b,cCD18 nos animais controles e infectados, sem diferencas qualitativas e quantitativas no figado...