Development of monoclonal antibodies against oropouche virus and its applicability to immunohistochemical diagnosis.

Orthobunyavirus oropouche ense virus (OROV), the causative agent of Oropouche fever, is widely dispersed in Brazil and South America, causing sporadic outbreaks. Due to the similarity of initial clinical symptoms caused by OROV with other arboviruses found in overlapping geographical areas, differential diagnosis is challenging. As for most neglected tropical diseases, there is a shortage of reagents for diagnosing and studying OROV pathogenesis. We therefore developed and characterized mouse monoclonal antibodies and, one of them recognizes the OROV nucleocapsid in indirect immunofluorescent (IFA) and immunohistochemistry (IHC) assays. Considering that it is the first monoclonal antibody produced for detecting OROV infections, we believe that it will be useful not only for diagnostic purposes but also for performing serological surveys and epidemiological surveillance on the dispersion and prevalence of OROV in Brazil and South America.

Serological evidence of dengue fever and its associated factors in health facilities in the Borena Zone, South Ethiopia.

BACKGROUND: Dengue fever (DF) is a re-emerging public health threat in Ethiopia. Yet, little is known about the epidemiology and risk factors of dengue infection in the region. In this study, the seroprevalence and associated risk factors of dengue virus infection were assessed in the Borena Zone health facilities. METHODS: A hospital-based cross-sectional study was conducted from July to August 2016. A total of 519 consecutive acute febrile patients attending the outpatient departments of Teltelle Health Center, Yabello and Moyale Hospital were enrolled. Data on socio-demographic and environmental risk factors were collected using a structured questionnaire. Three to five milliliter blood samples were collected from all participants and screened for dengue virus exposure using indirect immunofluorescent assay. RESULTS: The overall prevalence of anti-DENV IgG and IgM was 22.9% and 7.9%, respectively. DF serostatus was influenced by gender (adjusted odd ratio (AOR)=1.72; 95% CI 1.01-2.94), place of residence (AOR=2.69; 95%CL 1.55-4.64) that had a higher rate of exposure and recalling of a recent mosquito bite (AOR=2.98; 95% CI 1.51-5.89) probably imply recent and/or ongoing active transmission. CONCLUSION: This study showed that DF could potentially emerge as a public health threat in the study area. In addition to that, the observed low awareness of participants underlines the urgent need for further community-based studies to determine the environmental, and host factors that determine the extent of exposure to dengue virus infection in the area for appropriate control and prevention planning.

Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazil's northern Pantanal wetland / Soroprevalência, análise espacial e fatores de risco da infecção por Neospora caninum em bovinos no Pantanal norte do Brasil

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals ≤ 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study.

Resumo Neosporose é causada por um protozoário intracelular obrigatório, Neospora caninum. É considerada uma das causas mais comuns e frequentes de aborto em bovinos em todo o mundo. Para avaliar a prevalência de anticorpos anti- N. caninum e fatores de risco associados, amostras de soro foram coletadas de 2.452 bovinos em 262 fazendas no Pantanal norte, estado de Mato Grosso, Brasil. Cada fazendeiro preencheu um questionário para posterior análise dos dados epidemiológicos. Anticorpos anti-N. caninum foram detectados por meio da reação de imunofluorescência indireta (RIFI), utilizando um ponto de corte de 1:100. A prevalência total de anticorpos anti-N. caninum foi de 25,44% (Intervalo de Confiança - IC 95%; 20,10%; 30,78%) e a prevalência por rebanho foi de 76,72% (IC 95%; 71,60%; 81,84%). A presença de cães, a ocorrência de abortamento em vacas e a venda de bovinos para reprodução estiveram estatisticamente associadas à soropositividade em rebanhos, enquanto, o risco de fêmeas serem soropositivas para N. caninum foi maior em animais com idade ≤ 6 anos e na presença de cães. A análise espacial indicou que o risco relativo da doença é espacialmente constante e que as fazendas com maior prevalência de anticorpos anti-N. caninum estão localizados ao sul da região em estudo.

Leptospirosis in Caspian Sea littoral, Gilan Province, Iran.

In Iran, leptospirosis is endemic to Caspian Sea littoral. The disease appears as a seasonal infection mostly affecting people in rural areas involved in farming. We investigated the prevalence of this infection among suspected patients in Gilan Province by an indirect immunofluorescent assay (IFA), and two PCR protocols, a nested-PCR and a real-time PCR (qPCR), targeting rrs and lipL32 genes, respectively. We also identified the common Leptospira species by sequencing a partial sequence of rrs gene. Out of the 128 sera examined by IFA, 25.78% were positive with the antibody titers ≥1/80. The antibody titer in 39.06% of sera ranged from 1/10 to 1/140, and 35. 16% showed no antibodies, all considered negative. Nested PCR and qPCR detected Leptospira DNA in 20.31% and 18.75% of the sera, respectively. The two PCR assays had 98.43% agreement (K = 0.93) and showed an inverse correlation with the IFA titers. Also, three pathogenic Leptospira species, L. kirschneri (n = 10), L. introgans (n = 8), and L. borgpetersenii (n = 2) were identified from the clinical specimens in the study area. In our hands both PCR assays proved very efficient for early diagnosis of illness and could be used in combination with IFA for both diagnosis and epidemiological studies, but nested PCR was cheaper and appeared more appropriate for our laboratories in rural settings.

Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazils northern Pantanal wetland

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study.

Resumo Neosporose é causada por um protozoário intracelular obrigatório, Neospora caninum. É considerada uma das causas mais comuns e frequentes de aborto em bovinos em todo o mundo. Para avaliar a prevalência de anticorpos anti- N. caninum e fatores de risco associados, amostras de soro foram coletadas de 2.452 bovinos em 262 fazendas no Pantanal norte, estado de Mato Grosso, Brasil. Cada fazendeiro preencheu um questionário para posterior análise dos dados epidemiológicos. Anticorpos anti-N. caninum foram detectados por meio da reação de imunofluorescência indireta (RIFI), utilizando um ponto de corte de 1:100. A prevalência total de anticorpos anti-N. caninum foi de 25,44% (Intervalo de Confiança - IC 95%; 20,10%; 30,78%) e a prevalência por rebanho foi de 76,72% (IC 95%; 71,60%; 81,84%). A presença de cães, a ocorrência de abortamento em vacas e a venda de bovinos para reprodução estiveram estatisticamente associadas à soropositividade em rebanhos, enquanto, o risco de fêmeas serem soropositivas para N. caninum foi maior em animais com idade 6 anos e na presença de cães. A análise espacial indicou que o risco relativo da doença é espacialmente constante e que as fazendas com maior prevalência de anticorpos anti-N. caninum estão localizados ao sul da região em estudo.

Leishmaniasis tegumentaria: sensibilidad de la inmunofluorescencia indirecta en pacientes con diagnótico parasitológico / Tegumentary leishmaniasis: sensitivity of the indirect immunofluorescent assay in patients with parasitológical diagnosis

La leishmaniasis tegumentaria es una zoonosis endémica de zonas tropicales y subtropicales del mundo, de difícil diagnóstico laboratorial. En áreas endémicas se realiza el diagnóstico de presunción basado en la presentación clínica y la reactividad a la prueba cutánea de la leishmanina. Sin embargo, un diagnóstico definitivo de la enfermedad requiere la combinación de diferentes métodos, entre los cuales la inmunofluorescencia indirecta (IFI) es uno de los más utilizados. Para determinar la sensibilidad de la IFI, se sometió un total de 95 pacientes con lesiones comprobadas de leishmaniasis a las pruebas de leishmanina, biopsia histopatológica, cultivo e IFI. En 80 pacientes se logró el diagnóstico parasitológico a través de la histopatología y/o el cultivo. De ellos, 58 muestras (72,5%) dieron reacción positiva en la IFI para Leishmania, aunque 12 de ellas (21%) también mostraron serología positiva para enfermedad de Chagas. La sensibilidad de la IFI para los casos de leishmaniasis mucosa solamente fue 81,3% (n=16). Se concluye que la IFI es un método de moderada sensibilidad para el diagnóstico de leishmaniasis tegumentaria.

Tegumentary leishmaniasis is an endemic zoonosis of tropical and subtropical areas of the world. Diagnosis of this illness is difficult even using laboratorial methods. In endemics areas a presumptive diagnosis is carried out based on the clinical presentation and reactivity to the skin test. However, a definitive diagnosis of the illness requires the combination of different methods, such as the indirect immunofluorescence assay (IIF), one of the most used methods. In order to determine the sensitivity of the IIF, a total of 95 patients with proven lesions of leishmaniasis were subjected to skin test, histopathology, culture and IIF. Parasitological diagnosis was achieved in 80 patients through histopathology and/or culture. Fifty­eight samples of them (72.5%) gave positive reaction by the IIF although 12 of them (21%) also showed positive serology for Chagas disease. The sensitivity of the IIF for mucosal leishmaniasis only was 81.3% (n=16). We concluded that IIF has moderate sensitivity for the diagnosis of tegumentary leishmaniasis.