Detecção sorológica de anticorpos anti-Leishmania infantum chagasi em cães atendidos em hospital veterinário escola

Leishmaniasis is a zoonosis that currently ranks among the six endemics considered priority in the world. In Brazil, the geographic distribution of this disease increases every year, in dogs as well as in the human population. The State of Santa Catarina was considered area without previous cases for visceral leishmaniasis until 2010 when the first autochthonous cases were described in canines in the capital of the state. Based on the recent case in the State, the objectives of this study were to determine the percentage of serum sera in anti-Leishmania infantum antibodies among those seen at the University Veterinary Hospital in Canoinhas, Santa Catarina State, Brazil and to verify the owners’ knowledge about aspects of transmission, diagnosis and prophylaxis of canine visceral leishmaniasis. The animals were submitted to a physical examination and then a blood sample was collected without anticoagulant, the serum being obtained by centrifugation. Serum antibody detection was done by the Indirect Enzyme-linked immunosorbent assay (SNAP Leishmania® Idexx) and Dual Plate Platform (DPP® Biomanguinhos/Fiocruz) tests. Simultaneously, the owners answered a questionnaire to determine the socioeconomic profile and knowledge about visceral leishmaniasis. Although the veterinary hospital receives animals from several cities in the northern region of Santa Catarina and even southern Paraná, no animal among the 72 examined between June and December 2016 was reagent to the rapid tests. Most of the owners were unaware of the existence of the disease and had no information about the means of transmission and its consequences. It is concluded that although the tested animals are not serum reagents, it is necessary a broader communication of the disease, since at the moment it is common the transit of animals between regions, either by the trade or change of domicile of the owners. Leishmaniasis is an important zoonosis that is spreading every day, mainly because of the difficulties inherent in your vector control. It is the responsibility of veterinarians, according to the concept of unique health, search for cases and to publish information on how to prevent this zoonotic disease. Keywords: Visceral Leishmaniasis. Dogs. Immunochromatographic test. Enzyme immunoassay.A leishmaniose é uma zoonose que atualmente encontra-se entre as seis endemias consideradas prioritárias no mundo. No Brasil, a distribuição geográfica desta doença vem ampliando, tanto na população de cães, quanto na humana. O estado de Santa Catarina era considerado indene para leishmaniose visceral até 2010 quando foram descritos os primeiros casos autóctones em caninos na capital. Com base na recente detecção de casos no Estado, os objetivos deste trabalho foram determinar o percentual de cães soro reagentes na pesquisa de anticorpos anti-Leishmania infantum entre os atendidos no hospital veterinário da Universidade do Contestado, em Canoinhas, Santa Catarina e verificar o conhecimento dos tutores destes animais com relação a aspectos de transmissão, de diagnóstico e de profilaxia da leishmaniose visceral canina. Para tal, os animais foram submetidos a um exame físico e coleta de amostra de sangue sem anticoagulante, sendo o soro obtido por centrifugação. A detecção de anticorpos no soro foi pelos testes de ELISA indireto (SNAP Leishmania® Idexx) e imunocromatográfico (Dual Plate Platform - DPP® Biomanguinhos/Fiocruz). Simultaneamente os tutores respondiam a um questionário para determinar o perfil socioeconômico e o conhecimento sobre leishmaniose visceral. Dentre os 72 cães examinados entre junho a dezembro de 2016 nenhum foi reagente nos testes. Os animais atendidos residiam em diversas cidades da região norte de Santa Catarina, principalmente em Canoinhas e Três Barras e, até no sul do Paraná. Relativo aos tutores, a maioria relatou desconhecer sobre a existência da doença, sobre os meios pelos quais é transmitida e as suas consequências. Conclui-se que embora os animais testados não tenham sido reagentes, é necessária uma maior divulgação desta doença, pois atualmente é comum o trânsito de animais entre regiões, seja pelo comércio ou mudança de seus tutores. A leishmaniose é uma importante zoonose que vem se disseminando a cada dia, principalmente devido às dificuldades inerentes ao controle de seu vetor. Cabe aos médicos veterinários, de acordo com o conceito de saúde única, a busca ativa e precoce de casos e a divulgação de medidas profiláticas e de controle desta zoonose. Palavras-Chave: Leishmaniose Visceral. Cães. Teste imunocromatográfico. Ensaio imunoenzimático

PREVALÊNCIA DA LEISHMANIOSE VISCERAL EM CANINOS DA ÁREA URBANA DE MARACANAÚ, CEARÁ, BRASIL

A leishmaniose visceral (LV) é uma antropozoonose que se apresenta como um problema de saúde pública mundial. É causada pelo protozoário do gênero Leishmania e envolve o homem como hospedeiro definitivo e caninos como reservatórios no ambiente urbano. Caninos, quando infectados, são responsáveis pela transmissão que ocorre principalmente através da picada de flebotomíneos do gênero Lutzomyia. A LV é uma doença de caráter endêmico no Brasil, na década de 2000, aproximadamente 48% dos casos ocorreram na região Nordeste. Do total dos 184 municípios cearenses, 16 municípios apresentam transmissão intensa da doença, dentre eles, Maracanaú, localizado na Região Metropolitana de Fortaleza e o quarto município do estado do Ceará em ocorrência da leishmaniose visceral humana (LVH). Por se tratar de uma área de transmissão intensa para LVH, e no intuito de aprimorar o entendimento da leishmaniose visceral canina (LVC) no município, o objetivo deste estudo foi avaliar a soroprevalência em caninos domiciliados na zona urbana de Maracanaú, Ceará. Estudo seccional por meio de inquérito censitário canino de base populacional para identificação da prevalência de anticorpos para infecção por L. infantum, realizado pelo Centro de Controle de Zoonoses (CCZ), durante o período de 2013 a 2015. O inquérito envolveu 8.382 caninos em 4.603 domicílios no município de Maracanaú nos bairros: Grande Pajuçara, Jereissati I, Jereissati II, e Timbó. Todos os caninos existentes no domicílio foram testados pelo teste rápido de triagem imunocromatográfico (DPP), sendo que na positividade deste teste seria realizado o teste confirmatório de ensaio imunoenzimático (ELISA). Foram recolhidos e eliminados os caninos sororreagentes ao teste de ELISA, conforme prevê o programa nacional de controle da LV. Dos caninos incluídos neste estudo, 324 (4%) foram positivos no teste DPP e 46 caninos reagentes ao teste ELISA (0,5%). Nos estudos no estado do Ceará com as mesmas técnicas de diagnóstico do presente relato a prevalência foi de 5,7% no ano de 2016. Em Brasília (DF) e Patos (PB) as prevalências foram de 9,19% e 11,33%, respectivamente. A LV é tradicionalmente uma doença de caráter rural, entretanto, tem sido observada nas últimas décadas uma tendência de alteração no seu padrão pelo processo de expansão nas áreas urbanas. Nos anos 1970, Maracanaú sofreu grande transformação quando foi escolhido para sediar o Distrito Industrial de Fortaleza modificando o ambiente através da ocupação das áreas no entorno deste Distrito. Tais modificações ambientais favoreceram a transição epidemiológica da LV do meio rural para o urbano. A prevalência da LV em caninos foi aquém daquela observada em outros estudos similares, podendo ser atribuído, pelo menos parcialmente, as ações governamentais de controle vetorial e diagnóstico e controle da infecção entre caninos

Dengue Incidence and Climatic Factors

Introduction: This study was carried out to analyze the temporal evolution of the incidence of dengue in Araraquara-SP, Brazil, from 2012 to 2016, correlating the incidence with temperature and pluviometry. Methods: It was a cross-sectional design. The monthly number of cases from 2012 to 2016 was collected in the datafile of the Special Health Service of Araraquara-SESA of USP. Climatic variables related to temperature and rainfall were obtained from the websites of Agritempo and Department of Water and Electric Power, respectively. Statistical planning included incidence rates, and the tendency from 2012 to 2016, and it was studied the correlation between the number of dengue cases and each one of the climatic variables, considering the time-lag concept. Results: The incidence rates were 52.68, 376.52, 737.39, 3,660 and 809.48 per 100,000 inhabitants. Significant correlations were observed between the number of dengue cases and climatic variables after 2-4 months for high temperature and after 1-4 months for rainfall. Conclusions: The city faced an epidemic of dengue in 2015. It was identified the time lag in which hot weather and rain favored the occurrence of new cases: 2 to 4 months later for high temperature, and 1 to 4 months later for rainfall

Comparative assessment of skin reactivity to thimerosal- or phenol-preserved Imunoleish® antigen in dogs with suspected American Tegumentary Leishmaniasis in an endemic area of the state of Rio de Janeiro, Brazil

The leishmanin skin test (LST), which is an in vivo test that assesses the cellular immune responses to Leishmania-derived antigens, is an important tool in the laboratory diagnosis of American tegumentary leishmaniasis (ATL). This study aimed to compare the results obtained in LST employing the Imunoleish® antigen preserved with thimerosal (AgT) or phenol (AgP) and serological techniques to detect a possible infection caused by Leishmania (Viannia) braziliensis in dogs. The study included 172 dogs from an area endemic for ATL in the municipality of Paracambi, state of Rio de Janeiro, Brazil. The results obtained with Imunoleish® antigen preserved with thimerosal (AgT) or phenol (AgP) and serological tests were compared. Each dog received, intradermally, 0.1 mL of each antigen on the inner side of the right (AgT) and left (AgP) thighs. Five (2.7%) dogs presented ATL lesions. Of these, two were reactive to both formulations and three were reactive only to AgT. Among the 172 dogs, 68 (39.5%) were reactive only to AgT, 16 (9.3%)  only to AgP, and 11 (6.4%) to both formulations. Twenty-one (12.2%) sera samples were reactive by immunofluorescent antibody test (IFAT) and 21 enzyme-linked immunosorbent assay (ELISA). However, in only two dogs out of the five which Leishmania was isolated from, serological tests were positive. The LST and serological tests could be a useful tool in the diagnosis of L. (V.) braziliensis infection in dogs. Standardization of the techniques and reagents used could allow comparative studies on sensitivity, specificity, and positive and negative predictive values in dogs from different regions.Keywords: American Tegumentary Leishmaniasis, Leishmanin skin test, Diagnosis, Dogs, Host

Assessment of Renal Functions and Lesions in Dogs with Serological Diagnosis of Canine Visceral Leishmaniasis

Background: Visceral leishmaniasis is a complex vector-borne disease caused by the protozoan Leishmania infantum. In urban centers of South America, where this zoonotic cycle occurs, dogs seem to be the main reservoirs and infection sources. Animals with canine visceral leishmaniasis (CVL) may have a wide clinical spectrum, and dogs are usually classified as asymptomatic, oligosymptomatic, and symptomatic. Several organs are affected in canine CVL, and renal involvement is often a determining factor in dog prognosis. Nevertheless, serum markers are slow to indicate loss of renal function. The aim of this studywas to evaluate kidney impairment in dogs diagnosed with CVL.Material, Methods & Results: Blood and urine samples were collected from 45 dogs from Barra Mansa-RJ, and used for urinalysis, urine protein/creatinine (UPC) ratio, and serum concentrations of urea and creatinine. The animals were classified as symptomatic (42.2%), oligosymptomatic (37.8%), and asymptomatic (20.0%). Some alterations were found in the urine samples; pale-yellow color in 17.8%, low specific gravity in 6.7%, turbidity in 51.1%, proteinuria in 80%, occult blood in 46.7%, bilirubin in 8.89%, and glucose in 6.7% of the samples. According to the UPC ratio, 60% of dogs were proteinuric, and UPC > 2.0 was high in symptomatic dogs. Azotemia was observed only in three dogs with CVL.Discussion: The majority of dogs presented one or more symptoms of CVL, as expected in an endemic area from Brazil. Pale-yellow urine was observed in some samples, and this change, when accompanied by the decreased urine specific gravity in dogs with CVL, suggests some degree of kidney disease. The presence of epithelial and red blood cells, leukocytes, bacteria, suspended mucus, and phosphate crystals that precipitate in alkaline urines could be associated, to some degree, with the urine turbidity found in the present study. The alkaline urine identified in some dogs could be related to the animals’ diet, but renal tubular acidosis (RTA) is another possible cause when referring to animals with CVL. The abnormal presence of bilirubin and glycosuria can be justified by liver damage and glomerular and tubular damage, respectively. Occult blood was found in the urine of almost half of the tested dogs, which occurred because of the presence of red blood cells in the urine sediment and hematuria in some animals, could be caused by tubular and glomerular lesions. The presence of granular and hyaline casts found in the samples reinforce the possibility of tubular injury. We found different levels of proteinuria; it was an important result, possibly caused by immune complex deposition in addition to tubular disease. Most tested dogs, including animals without clinical manifestation, were classified as proteinuric or borderline proteinuric, showing that the renal disease could be the only clinical manifestation of CVL and that it could progress from slight proteinuria to end-stage renal disease, resulting in chronic renal failure, which is the main cause of death. The UPC ratio > 2.0 was significantly the more frequent finding in this study, mainly in symptomatic dogs. This result indicates a glomerular disease in these animals, reinforcing that the progression of renal disease follows the clinical progression of CVL. A few serum samples showed increased urea and creatinine levels, proving that azotemia is an uncommon finding in CVL-infected dogs. In conclusion, urinalysis helped in the early identification of renal injury in CVL-infected dogs, highlighting elements that reinforce the presence of tubular or glomerular lesions, or both, even in non-azotemic dogs. The high frequency of symptomatic dogs with UPC ratio > 2.0 suggests a relationship between the progression of renal disease and the clinical progression of CVL

IDENTIFICATION OF CANINE VISCERAL LEISHMANIASIS IN A PREVIOUSLY UNAFFECTED AREA BY CONVENTIONAL DIAGNOSTIC TECHNIQUES AND CELL-BLOCK FIXATION

After the report of a second case of canine visceral leishmaniasis (CVL) in São Bento da Lagoa, Itaipuaçu, in the municipality of Maricá, Rio de Janeiro State, an epidemiological survey was carried out, through active search, totaling 145 dogs. Indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), and rapid chromatographic immunoassay based on dual-path platform (DPP(r)) were used to perform the serological examinations. The parasitological diagnosis of cutaneous fragments was performed by parasitological culture, histopathology, and immunohistochemistry. In the serological assessment, 21 dogs were seropositive by IFA, 17 by ELISA, and 11 by DPP(r), with sensitivity of 66.7%, 66.7% and 50%, and specificity of 87.2%, 90.2% and 94%, respectively for each technique. The immunohistochemistry of bone marrow using the cell-block technique presented the best results, with six positive dogs found, three of which tested negative by the other parasitological techniques. Leishmania sp. was isolated by parasitological culture in three dogs. The detection of autochthonous Leishmania infantum in Itaipuaçu, and the high prevalence of seropositive dogs confirm the circulation of this parasite in the study area and alert for the risk of expansion in the State of Rio de Janeiro

Cryptosporidium spp. in Aratinga jandaia, Diopsittaca nobilis and Pionus menstruus in Brazil

Background: Cryptosporidium is an important protozoan in public health and veterinary medicine that often causes diarrhea in an array of hosts in developed/developing countries. Infection of the gastrointestinal system is the most common, but the respiratory system and other sites can also be affected, especially in birds and immunocompromised individuals. Transmission occurs through ingestion or inhalation of oocysts. The number of wild animals, including those in the class of birds, infected with this parasite has grown in recent years. This study aimed to report parasitism by Cryptosporidium spp. in captive-raised birds of family Psittacidae at the Rio City Zoo in Rio de Janeiro, Brazil.Materials, Methods & Results: Thirty-three pools of fecal samples of the species Amazona aestiva, Amazona amazonica, Anodorhynchus hyacinthinus, Ara auricollis, Ara canga, Ara glaucogularis, Ara macao, Ara manilapa, Ara maracana, Ara rubrogenys, Aratinga erythrogenys, Aratinga cactorum, Aratinga auerea, Aratinga mitrata, Aratinga auricapilla, Aratinga jandaia, Aratinga wagleri, Aratinga leucophthalmus, Brotogeris acuticaudata, Cynoliseus patagonus, Caracopsis vasa, Diopsittaca nobilis, Graydidascalus brachyurus, Muopsitta monachus, Nangayus nenday, Pionites melancephala, Pionites leucogaster, Pionus menstruus, Pionus chalcopteus, Pionus maxiliani, Pyrrhura perlata, Pyrrhura leucotis, and Triclharia malachitacea, kept in separate enclosures, were analyzed using Enzyme-linked Immunosorbent Assay (ELISA) for detection of parasitic antigens. Quantitative Polymerase Chain Reaction (qPCR) was conducted in order to identify the species Cryptosporidium in the positive samples targeting the small subunit ribosomal RNA gene (SSU rRNA), followed by sequencing and analysis of the DNA amplicons. Cryptosporidium spp. antigen was detected in three (9%) of the thirtythree pools assessed, corresponding to the following species of family Psittacidae: Jandaya parakeet (Aratinga jandaya), Red-shouldered macaw (Diopsittaca nobilis), and Blue-headed parrot (Pionus menstruus). Positivity of the three samples was confirmed by qPCR analysis, but it was not possible to identify the species of Cryptosporidium by this technique.Discussion: Zoonotic diseases, such as cryptosporidiosis, have been reported in wild and captive-bred animals worldwide. Several species of the class of birds are parasitized by Cryptosporidium spp. Infection in order Psittaciformes has been described in some species raised in captivity in Brazil; however, no reports of Cryptosporidium spp. infecting the following species of order Psittaciformes: Jandaya parakeet (Aratinga jandaia), Red-breasted macaw (Diopsittaca nobilis), and Blue-headed parrot (Pionus menstrus), were found in the specific scientific literature. The present study detected infection by Cryptosporidium spp. in three species of order Psittaciformes using ELISA and confirmed the positivity of the samples by qPCR, but the species of Cryptosporidium could not be identified. These infected birds should be carefully investigated, with identification of the species and analysis of their zoonotic potential, because they can be sources of environmental contamination and infection for the caregivers, visitors, and other animals of the Zoo. This is the first report of parasitism by Cryptosporidium spp. in these species of family Psittacidae in Brazil

Sensitivity and specificity of parallel or serial serological testing for detection of canine Leishmania infection

In Brazil, human and canine visceral leishmaniasis (CVL) caused byLeishmania infantum has undergone urbanisation since 1980, constituting a public health problem, and serological tests are tools of choice for identifying infected dogs. Until recently, the Brazilian zoonoses control program recommended enzyme-linked immunosorbent assays (ELISA) and indirect immunofluorescence assays (IFA) as the screening and confirmatory methods, respectively, for the detection of canine infection. The purpose of this study was to estimate the accuracy of ELISA and IFA in parallel or serial combinations. The reference standard comprised the results of direct visualisation of parasites in histological sections, immunohistochemical test, or isolation of the parasite in culture. Samples from 98 cases and 1,327 noncases were included. Individually, both tests presented sensitivity of 91.8% and 90.8%, and specificity of 83.4 and 53.4%, for the ELISA and IFA, respectively. When tests were used in parallel combination, sensitivity attained 99.2%, while specificity dropped to 44.8%. When used in serial combination (ELISA followed by IFA), decreased sensitivity (83.3%) and increased specificity (92.5%) were observed. Serial testing approach improved specificity with moderate loss in sensitivity. This strategy could partially fulfill the needs of public health and dog owners for a more accurate diagnosis of CVL

Tratamento cirúrgico associado à terapia antifúngica convencional na esporotricose felina

Esporotricose é causada pelo fungo Sporothrix schenckii, que infecta os humanos e animais. Os gatos domésticos apresentam elevado potencial zoonótico pela riqueza parasitária encontrada nas lesões cutâneas e a transmissão zoonótica pode correr principalmente através de contato com exsudatos de lesões, mordeduras ou arranhaduras de gatos. É relatado o caso de um gato com esporotricose que, após 19 meses de tratamento regular com itraconazol, apresentou uma nova lesão localizada na bolsa escrotal. Após 5 meses de tratamento, não havendo regressão da lesão, foi realizada a exérese da bolsa escrotal e orquiectomia. A terapia antifúngica foi mantida durante 2 meses após o procedimento cirúrgico. Três meses após a suspensão do itraconazol, o animal ainda permanece sem lesões. O objetivo deste estudo foi relatar a associação do tratamento cirúrgico à terapêutica convencional da esporotricose felina, após falência da mesma utilizada isoladamente. No caso relatado, a excisão cirúrgica da bolsa escrotal e orquiectomia representaram uma opção terapêutica em associação com agentes antifúngicos