Spatial analysis of canine leishmaniasis in an area of transmission of the semi-arid region of the State of Paraíba, Brazil / Análise espacial da leishmaniose canina em uma área de transmissão do semiárido paraíbano, Brasil

Abstract Visceral leishmaniasis is a widely distributed zoonosis and canine infection is an important indicator of risk for the occurrence of the disease in humans. The goal of this analysis was to study the spatial clustering of canine leishmaniasis (CL) in the municipality of Santa Luzia, state of Paraíba. For this, 749 samples of canine plasma were tested using three serological tests. The dog was considered positive if it reacted in two serological tests. The location of the residences was performed with a Global Positioning System receiver (GPS Garmin® eTrex 30), and used to perform georeferencing and spatial analysis. The prevalence of CL was 15.49% and it was observed that most cases of the urban area were concentrated in the Frei Damião neighborhood, on the outskirts of the city, where a high-risk cluster for the occurrence of the disease was formed (p = 0.02; RR = 2.48). No statistically significant cluster was observed in rural areas. CL is widely distributed in the municipality of Santa Luzia in a heterogeneous manner and with a tendency to urbanization. The areas identified with high prevalence and highest risk should be prioritized to maximize the efficiency of the Visceral Leishmaniasis Surveillance and Control Program and minimize the chance of new canine and human cases.

Resumo A leishmaniose visceral é uma zoonose amplamente distribuída, e a infecção canina é um importante indicador de risco para a ocorrência da doença em humanos. O objetivo foi estudar o comportamento espacial da leishmaniose canina (LC) no município de Santa Luzia, estado da Paraíba. Para tal, 749 amostras de plasma canino foram testadas por meio de três técnicas sorológicas. O cão seria considerado positivo se apresentasse duas sorologias reagentes. A localização das residências foi realizado pelo receptor de Sistema de Posicionamento Global (GPS Garmin® eTrex 30) e usado para a realização do georreferenciamento e análise espacial. A prevalência da LC foi de 15,49% e observou-se que a maioria dos casos de LC da zona urbana estava concentrada no bairro Frei Damião, localizado na periferia da cidade, onde se formou um cluster de alto risco para ocorrência da doença (p = 0,02; RR = 2,48). Nenhum cluster estatisticamente significativo foi observado na zona rural. A LC está amplamente distribuída no município de Santa Luzia, porém de forma heterogênea e com tendência à urbanização. As áreas identificadas com intensa prevalência e maior risco devem ser priorizadas para maximizar a eficiência do Programa de Vigilância e Controle da Leishmaniose Visceral e minimizar a chance de novos casos caninos e humanos.

Demographic and spatial study of visceral leishmaniasis in the state of Alagoas, Brazil, during 2007-2018

Abstract INTRODUCTION: Visceral leishmaniasis has a broad worldwide distribution and constitutes a public health problem in the Northeast of Brazil. Located in this region is the state of Alagoas, where the disease is endemic in humans and where there has been a significant increase in the number of positive dogs. The objective of this study was to describe the temporal and spatial distribution of the cases of human VL in the state of Alagoas with the aim of identifying transmission risk areas in the period from 2007 to 2018. METHODS: The data available in the National Disease Notification System (SINAN-NET) were used. The Bayesian incidence rate and the Moran's global index were calculated using the Terra View 4.2.2 program, and the maps were created using QGIS2.18.0. RESULTS: From the 102 municipalities, 68.6% (n= 70) had at least one notified case of VL in the years of study. A total of 489 cases were registered, with an average of 40.7 cases per year and an incidence rate of 1.25/100,000 inhabitants. The highest number of confirmed cases (105) occurred in 2018. Male individuals and children between 1-4 years old were the most affected, and 64% of the cases were in rural areas. Spatial dependence was detected in all the intervals except for the first triennium, and clusters were formed in the west of the state. CONCLUSIONS: Alagoas presented an accentuated geographical expansion of VL, and it is necessary to prioritize areas and increase surveillance actions and epidemiological control.

Natural Infection by Leishmania infantum in domestic cats (Felis catus) in a municipality of moderate transmission in the Brazilian semi-arid region.

The main clinical, anatomopathological, and molecular aspects of the infection by Leishmania infantum are described in two cats with multicentric cutaneous, nodular, and ulcerated lesions. The animals were submitted to a clinical examination, followed by serological, molecular and parasitological exams, with culture and isolation of the parasite, and subsequent isoenzymatic characterization. The animals were euthanized and necropsied. Case 1 was an adult, female, mixed-bred stray cat. Case 2 was an adult, male, mixed-bred and domiciled cat. Both were positive for the presence of anti-L. infantum antibodies. In the cytology of the cutaneous nodules and lymph nodes, amastigote forms of Leishmania spp. could be visualized, free and in the interior of the macrophages. In the histopathology, the lesions were characterized by nodular granulomatous and/or ulcerative dermatitis, associated to amastigote forms of Leishmania spp. By means of the polymerase chain reaction, the sequence of the L. infantum kDNA minicircle was amplified. It is concluded that the infection by L. infantum occurs in cats in the State of Paraíba, Northeast region of Brazil and the need to understand the immunological profile of the visceral leishmaniasis in the feline population is highlighted with aimed at the control measures in public health.

Clinical aspects and diagnosis of leishmaniasis in equids: a systematic review and meta-analysis / Aspectos clínicos e diagnóstico da leishmaniose em equídeos: uma revisão sistemática e meta-análise

Abstract Leishmaniases are a group of diseases of zoonotic importance caused by over 20 species of protozoa of the genus Leishmania, in which domestic dogs are considered to be the main reservoir for the disease. However, the involvement of other vertebrates as reservoirs for these parasites has also been investigated. Therefore, the objective of the present study was to carry out a systematic review with meta-analysis on occurrences of leishmaniasis in equids. The case reports described animals with cutaneous symptoms of leishmaniasis (papules, nodules, ulcers or crusts) that regressed spontaneously, located mainly on the head and limbs, from which three species of protozoa were identified in the lesions: Leishmania braziliensis, Leishmania infantum and Leishmania siamensis. In turn, the meta-analysis showed a combined prevalence of 25%, although with high heterogeneity among the studies, which was attributed to the use of different methods for diagnosing the disease. Leishmaniasis in equids is a benign disease but it should be included in the differential diagnosis of cutaneous diseases among these species. Seroepidemiological studies are important in investigating and monitoring suspected exposure of these hosts to the parasite, especially in endemic areas. However, there is also a need to standardize diagnostic methods.

Resumo As leishmanioses são um grupo de doenças de importância zoonótica causadas por mais de 20 espécies de protozoários do gênero Leishmania, sendo o cão doméstico considerado o principal reservatório da doença. No entanto, diversas pesquisas têm investigado o envolvimento de outros vertebrados como reservatórios do parasita. Portanto, o objetivo do presente estudo foi realizar uma revisão sistemática com meta-análise da ocorrência de leishmaniose em equídeos. Os relatos de caso descreviam animais com sintomas cutâneos de leishmaniose (pápulas, nódulos, úlceras, crostas) que regrediam espontaneamente, localizadas principalmente na cabeça e membros, sendo identificadas três espécies do protozoário nas lesões: Leishmania braziliensis, Leishmania infantum e Leishmania siamensis. Por sua vez, a meta-análise evidenciou uma prevalência combinada de 25%, porém com alta heterogeneidade entre os estudos, atribuída às diferenças nos métodos utilizados no diagnóstico da doença. A leishmaniose em equídeos é uma doença benigna, porém deve ser incluída no diagnóstico diferencial de doenças cutâneas nessas espécies. Os estudos soroepidemiológicos são importantes para investigar e monitorar a suspeita de exposição desses hospedeiros ao parasita, principalmente em áreas endêmicas, porém há necessidade de padronização dos métodos de diagnóstico.

Avaliação da composição e da contagem de células somáticas do leite de cabra cru do Cariri paraibano / Evaluation of composition and counting of somatic cells of cair raw of Cariri paraibano

A CCS é um método aceito internacionalmente para avaliar a sanidade da glândula mamária. Objetivou-se neste trabalho determinar a composição e a contagem de células somáticas no leite de cabra cru do Cariri paraibano proveniente quatro mini usinas denominadas A, B, C e D. Os resultados médios obtidos para os parâmetros físico-químicos foram 3,5% de gordura; 3,0% de proteína; 4,2% de lactose; 11,70% de sólidos totais; 8,2% de sólidos não gordurosos e 1,4 x 106 para CCS. Os resultados obtidos demonstram que dos componentes do leite apenas a lactose estava fora dos padrões e que a alta CCS reforça a necessidade de implementação dos programas de controle de qualidade para melhoria da sanidade da glândula mamária.

Seroprevalence and risk factors associated with canine visceral leishmaniasis in the State of Paraíba, Brazil

Abstract INTRODUCTION: The aim of this study was to determine the seroprevalence and risk factors associated with canine visceral leishmaniasis in Paraíba State. METHODS: Blood samples were collected from 411 dogs in four municipalities of Paraíba State. The seroprevalence was assessed by using ELISA. RESULTS: The seroprevalence ranged from 7.2% to 20%. The risk factors that were associated with the disease were the presence of ticks and contact with other animals. CONCLUSIONS: The seroprevalence of canine visceral leishmaniasis is high in Paraíba, indicating the need for health authorities to resume visceral leishmaniasis control activities. The ruralization of suburban areas in the region tends to homogenize the risk factors between suburban and rural areas.

Demographic and clinical characterization of human visceral leishmaniasis in the State of Pernambuco, Brazil between 2006 and 2015

Abstract INTRODUCTION: Human visceral leishmaniasis (HVL) primarily occurs in regions that present socioeconomic, health, and environmental vulnerability. In Pernambuco, Brazil, this neglected zoonosis has expanded in magnitude as well as geographically, and efforts to manage HVL have been insufficient to contain its spread. The objective of this study was to describe the epidemiological profile of human illness due to HVL during 2006 to 2015 in Pernambuco State. METHODS: This study was conducted using secondary data collected from the Health Information System. RESULTS: During the study period, Pernambuco accounted for 2.4% of HVL cases in Brazil, with 49.6% of these concentrated in the macroregions of São Francisco Valley and Araripe. The percentage of municipalities that reported cases at the beginning of the study period increased from 21.1% (n = 39) to 43.8% (n = 81) by the end of the period. Cases were found predominantly among males, brown-skinned individuals, children aged 1-4 years, and individuals with incomplete 1st to 4th grade education. Coinfection with HIV was present in 5.6% of cases. Incidence was 9.7 cases per 100,000 inhabitants, and lethality was 12.3%. CONCLUSIONS: HVL has shown worrying expansion and evolution, in addition to high lethality, in Pernambuco. The only study of its kind in the past decade, it was evident from this study that despite efforts to contain this disease, HVL in Pernambuco exhibits patterns similar to those described in previous studies. Based on our results, we suggest reassessing the current prevention and control measures in the state.

Perfil imunogênico de vacinas inativadas contra agalaxia contagiosa em ovinos / Immunogenic profile of inactivated vaccines against contagious agalactia in sheep

Este trabalho teve como objetivo avaliar o perfil imunogênico em ovinos de três vacinas produzidas com linhagens brasileiras de Mycoplasma agalactiae. O perfil proteico do antígeno vacinal foi avaliado por SDS-PAGE e a imunogenicidade da vacina pela técnica de Western blot. A vacina foi inativada com formol, adsorvida em hidróxido de alumínio (Vacina 1), Montanide IMS-2215 (Vacina 2), Montanide Gel-01 (Vacina 3) e administradas em três doses. Entre a primeira e a segunda dose houve um intervalo de 21 dias, e entre a segunda e a terceira de 180 dias. O pool de soros de dez ovinos coletados nos períodos 0, 21, 35, 90, 150, 210, 270 e 360 dias pós-vacinação foram testados pela técnica de Western blot. A vacina 2 foi mais antigênica, com detecção de anticorpos 21 dias após a primeira dose. Para as vacinas 1 e 3, os anticorpos são verificados após 35 dias, com queda acentuada aos 90 dias; apenas anticorpos contra a proteína de 48 kDa apareceram após a terceira dose de forma discreta. Contra a vacina 2, ainda persistiram anticorpos contra as proteínas de 48, 55 e 80 kDa nos períodos 90, 150 e 210, que aumentaram após a terceira dose. Conclui-se que a vacina 2 induziu a resposta humoral de forma estável contra proteínas de M. agalactiae.

This study aimed to evaluate the immunogenic profile in sheep of three vaccines produced with a field Brazilian strains of Mycoplasma agalactiae. The vaccine protein profile was evaluated by SDS-PAGE and vaccine immunogenicity by Western blot. The vaccine was inactivated with formaldehyde and adsorbed onto three different adjuvants: with aluminum hydroxide (Vaccine 1), Montanide IMS 2215 (Vaccine 2), and Gel Montanide-01 (Vaccine 3). The vaccine was administered in three doses. Between the first and second dose there was an interval of 21 days, and between the second and the third one of 180 days. A pool of ten sera collected in 0, 21, 35, 90, 150, 210, 270 and 360 days after first vaccination were tested by Western blot techniques. The second vaccine was more antigenic with antibody detection 21 days after first dose. For both vaccines 1 and 3, antibodies were present 35 days after first dose, with a significant drop at 90 days; only antibodies against 48 kDa protein discreetly appeared after the third dose. Stimulation induced by vaccine 2 produced antibodies against 48, 55 and 80 kDa proteins that persisted until 90, 150 and 210 days after first dose, which rose again after third dose. It was concluded that the vaccine 2 induced stable humoral immunity against M. agalactiae proteins.

Ehrlichia canis and Rickettsia spp. in dogs from urban areas in Paraiba state, northeastern Brazil / Ehrlichia canis e Rickettsia spp. em cães de áreas urbanas da Paraíba, nordeste do Brasil

Abstract The aims of our study was to identify Ehrlichia canis and antibodies against Rickettsia spp. belonging to the spotted fever group (SFG) in dogs sampled from Paraiba state, northeastern Brazil. Blood and serum samples collected by convenience from dogs in urban areas of five municipalities were analyzed by real-time PCR for the detection of E. canis DNA and by immunofluorescence assay test (IFAT) for the identification of antibodies against Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii and R. rhipicephali antigens. E. canis DNA was detected in 8.9% (64/719) of the blood samples, whereas 5.63% (43/763) of the serum samples were positive for at least one of the Rickettsia antigens tested by IFAT. This study showed for the first time the occurrence of E. canis and suggested the circulation of SFG Rickettsia in dogs in the study region of Paraiba state, northeastern Brazil.

Resumo Os objetivos do nosso estudo foram identificar Ehrlichia canis e anticorpos contra Rickettsia spp. pertencentes ao Grupo da Febre Maculosa (GFM) em cães amostrados no estado da Paraíba, nordeste do Brasil. As amostras de sangue e soro, coletados por conveniência, de cães em áreas urbanas de cinco municípios foram analisadas por PCR em tempo real para a detecção de DNA de E. canis e pela Reação de Imunofluorescência Indireta (RIFI) para identificação de anticorpos contra Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii e R. rhipicephali. O DNA de E. canis foi detectado em 8,9% (64/719) das amostras de sangue, enquanto que 5,63% (43/763) das amostras de soro foram positivas para pelo menos um dos antígenos de Rickettsia testados por RIFI. Este estudo mostrou pela primeira vez a ocorrência de E. canis e sugere a circulação de Rickettsia do GFM em cães na região em estudo do estado da Paraíba, Nordeste do Brasil.

Canine visceral leishmaniasis in the Northeast Region of Brazil

Visceral leishmaniasis (VL) is a zoonosis that affects dogs and other mammals, including humans. Contact with dogs is a major risk factor for humans. This disease is endemic in several regions of Brazil. The aim of this study was to determine the prevalence of Leishmania spp. infection in dogs and to correlate it with possible risk factors. Methods Blood samples were collected from 391 dogs of different ages, breeds, and both genders, coming from Campina Grande, Paraíba state, Brazil. An epidemiological questionnaire was employed in order to identify risk factors associated with the disease. Serological tests were performed using indirect immunofluorescence, enzyme-linked immunosorbent assay (ELISA S7®) and polymerase chain reaction. Results Leishmania spp. antibodies were detected in 33 (8.4 %) and 17 (4.3 %) dogs according to the indirect immunofluorescence test (IFAT) and enzyme-linked immunosorbent assay (ELISA S7®), respectively. PCR results indicated the presence of L. chagasi DNA in only eight (2 %) blood samples. There was a significant association between reactive animals and contact with animals from different houses (OR = 4.1; p= 0.02). Conclusions It is suggested that CVL may occur in urban areas. Moreover, it is demonstrated that the association among different diagnostic tests may lead to a more accurate identification of positive animals, which might help to improve the disease control and prevent euthanasia in false-positive results.

Canine visceral leishmaniasis in the Northeast Region of Brazil

Background Visceral leishmaniasis (VL) is a zoonosis that affects dogs and other mammals, including humans. Contact with dogs is a major risk factor for humans. This disease is endemic in several regions of Brazil. The aim of this study was to determine the prevalence of Leishmania spp. infection in dogs and to correlate it with possible risk factors. Methods Blood samples were collected from 391 dogs of different ages, breeds, and both genders, coming from Campina Grande, Paraíba state, Brazil. An epidemiological questionnaire was employed in order to identify risk factors associated with the disease. Serological tests were performed using indirect immunofluorescence, enzyme-linked immunosorbent assay (ELISA S7®) and polymerase chain reaction. Results Leishmania spp. antibodies were detected in 33 (8.4 %) and 17 (4.3 %) dogs according to the indirect immunofluorescence test (IFAT) and enzyme-linked immunosorbent assay (ELISA S7®), respectively. PCR results indicated the presence of L. chagasi DNA in only eight (2 %) blood samples. There was a significant association between reactive animals and contact with animals from different houses (OR = 4.1; p= 0.02). Conclusions It is suggested that CVL may occur in urban areas. Moreover, it is demonstrated that the association among different diagnostic tests may lead to a more accurate identification of positive animals, which might help to improve the disease control and prevent euthanasia in false-positive results.(AU)

Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil / Pesquisa de Ehrlichia canis, Babesia spp. e Hepatozoon spp. em cães de uma região semiárida do Brasil

This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100) of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR), respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100) of the animals. Babesia vogeli was detected by PCR in ten animals (10%) and was correlated with young age (p = 0.007) and thrombocytopenia (p = 0.01). None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.

O presente estudo avaliou a ocorrência de infecção por Ehrlichia spp., Babesia spp. e Hepatozoon spp. em 100 cães, infestados por carrapatos, oriundos de uma região semiárida do Estado da Paraíba, Nordeste do Brasil. Amostras de sangue e de carrapatos foram coletadas dos animais, e um questionário foi submetido aos proprietários dos cães para obter dados gerais. As amostras de sangue foram utilizadas para realização de hemograma, esfregaço sanguíneo e detecção molecular dos hemoparasitos. Os 1.151 carrapatos coletados foram identificados como Rhipicephalus sanguineus; os esfregaços sanguíneos revelaram mórulas sugestivas de E. canis em 4% (4/100) de cães fêmeas não vacinadas, e 34% e 25% dos cães foram positivos para Ehrlichia canis pela imunofluorescência indireta (IFI) e reação em cadeia pela polimerase (PCR), respectivamente. Os esfregaços sanguíneos revelaram merozoítas sugestivas de Babesia em eritrócitos de 2% (2/100) dos animais. Babesia vogeli foi detectada por PCR em dez animais (10%) e foi correlacionada com a idade jovem (p=0,007) e trombocitopenia (p=0,01). Nenhum dos animais apresentou positividade para Hepatozoon spp. Esses resultados indicam que E. canis é o principal patógeno canino transmitido por carrapato, na área estudada, e fornece o primeiro relato de infecção por B. vogeli em cães do Estado da Paraíba.

Isolamento de clones imunoreativos e caracterização in silico de novos antígenos recombinantes de Leishmania chagasi / Isolation of immunoreactive clones and characterization in silico of new recombinant antigens of Leishamania chagasi

As leishmanioses são endêmicas em mais de 88 países e representam um sério problema de saúde pública no Brasil. O diagnóstico precoce,o controle vetorial e,até recentemente,a eliminação de cães soropositivos, constituem a única abordagem para a luta contra a leishmaniose visceral.O sorodiagnóstico humano e canino, contudo,é dificultado por reações cruzadas com outras doenças e pelas dificuldades na padronização do antígeno para a produção de testes em larga escala para os programas de saúde pública.Antígenos recombinantes podem potencialmente contornar ambos os problemas e contribuir para uma redução do custo de produção.Atualmente apenas 2 antígenos recombinantes estão no mercado para o diagnóstico do calazar.Patentes concedidas...pelo calazar no Mundo podem restringir de forma severa as possibilidades de desenvolvimento de um teste nacional livre de pagamentos de royalties pelo seu uso em programas de saúde pública, tanto para o diagnóstico como para a vacinação e a imunoterapia.Neste trabalho descrevemos o isolamento e a caracterização de 10 novos antígenos reconhecidos pelos anticorpos séricos de humanos ou cães com calazar.A triagem de uma biblioteca de cDNA de Leishmania chagasi com soros previamente imunoadsorvidos contra um fragmento de HSP70 recombinante permitiram o isolamento de clones expressando HSP83,K39 e novos antígenos,pertencentes a 2 categorias:proteínas hipotéticas ou proteínas com função conhecida,podendo ser enzimas ou proteínas estruturais; exceto por um caso,as proteínas não haviam sido previamente descritas como antígeno nas leishmanioses. Não houve antígenos comuns às 2 triagens,exceto os antígenos imunodominantes HSP83 e K39.Alguns dos 10 novos antígenos têm repetições de aminoácidos curtas ou longas, que podem sugerir um mecanismo para conferir a antigenicidade nestas proteínas.Domínios e motivos puderam ser identificados em alguns destes antígenos, assim com funções prováveis atribuídas e localização intracelular.Os epitopos...