Avaliação da capacidade adjuvante do cloreto de dimetildioctadecilamônio associado ao hidróxido de alumínio na indução da resposta imune humoral de bovinos vacinados com o vírus da diarréia viral bovina

The humoral immune response of cattle vaccinated with inactivated bovine viral diarrhea virus (BVDV) using the association of dimethyl dioctadecyl ammonium chloride (DDA-chloride) and aluminum hydroxide (vaccine B) as adjuvant, was compared with the same antigenic formulation adsorbed with only aluminum hydroxide (vaccine A). The neutralizing antibodies titers were assessed two weeks after the second vaccine dose. The animals that received vaccine B had better humoral responses than the calves vaccinated with vaccine A. The mean titers of neutralizing antibodies, expressed in Log2, for the calves that received vaccine B was higher to those observed in group vaccinated with vaccine A. These results show that the inclusion of DDA chloride in vaccine formulations adsorbed with aluminum hydroxide determine a better humoral immune response in cattle vaccinated with BVDV inactivated.A resposta imunológica humoral de bovinos vacinados com o vírus da diarréia viral bovina (BVDV) inativado, tendo como adjuvante o cloreto de dimetildioctadecilamônio (DDA cloreto) associado ao hidróxido de alumínio (vacina B), foi comparada com uma vacina contendo o mesmo antígeno adsorvido apenas com hidróxido de alumínio (vacina A). Duas semanas após a segunda dose foi avaliado o título de anticorpos neutralizantes dos animais que receberam as duas preparações de antígenos. Os animais que receberam a vacina B apresentaram melhor resposta imune humoral quando comparados com os animais vacinados com a vacina A. O título médio de anticorpos neutralizantes, expresso em Log2, dos animais que receberam a vacina B foi superior (P<0,05) ao observado no grupo vacinado com a vacina A. Esse resultado demonstra que, em bovinos vacinados com o BVDV inativado, a inclusão do DDA cloreto em formulações de vacinas adsorvidas com hidróxido de alumínio potencializa a resposta imune humoral

Perfil plasmidial de Staphylococcus hyicus isolados de suínos com epidermite exsudativa no Brasil

Staphylococcus hyicus isolados de leitões com lesões de pele foram estudados no que se refere aos seus plasmídios e resistência a agentes antimicrobianos. Plasmídios de diferentes pesos moleculares foram detectados nas seis amostras de Staphylococcus hyicus. Todas as amostras mostraram resistência à tetraciclina e um pequeno plasmídio de 4.0 Kb, com uma exceção. Experimentos de cura e transformação demonstraram que este plasmídio abriga o determinante genético para resistência à tetraciclina nas amostras estudadas

Risk factors for bovine herpesvirus 1 infection in cattle herds in the West region of Parana State

O objetivo deste estudo foi determinar a prevalência de rebanhos positivos (focos) e identificar os fatores de risco que possam estar associados com a infecção pelo herpesvírus bovino 1 (BoHV-1) em rebanhos bovinos com atividade reprodutiva, na região Oeste do Estado do Paraná. O delineamento estatístico, amostras de soro e informações referentes às propriedades foram as empregadas para o estudo da brucelose bovina no Estado do Paraná dentro do contexto do Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose. Foram avaliadas 1930 fêmeas com idade igual ou superior a 24 meses, provenientes de 295 rebanhos não vacinados contra o BoHV-1. Para o diagnóstico sorológico da infecção pelo BoHV-1, foi utilizado um ensaio imunoenzimático (ELISA) indireto. Em cada propriedade foi aplicado um questionário epidemiológico, afim de obter informações epidemiológicas e práticas de manejo empregadas. Dos 295 rebanhos analisados, 190 foram considerados positivos para o BoHV-1, com a prevalência de rebanhos de 64,41% (I.C.95% = 58,65-69,87%). As variáveis consideradas fatores de risco para a infecção pelo BoHV-1 na análise de regressão logística multivariada foram: i) número ('> OU ='23) fêmeas com idade '> OU ='24 meses (OR=2,22; IC: 1,09-4,51); ii) compra de reprodutores (OR=2,68; IC: 1,48-4,82); iii) uso de pastagens comuns (OR=5,93; IC: 1,31-26,82); iv) histórico de abortamento nos últimos 12 meses (OR=2,37; IC: 1,09-5,16); v) presença de animais silvestres (OR=8,86; IC: 1,11-70,73). Estes resultados indicam que a infecção pelo BoHV-1 está amplamente distribuída na região estudada e que fatores relacionados às características das propriedades e ao manejo estão associados à infecçãoThe aim of this study was to determine the prevalence of positive herds and to identify the risk factors for BoHV-1 infection in breeding cattle herds in the Western region of Parana State. The statistic delineation, the serum samples and information regarding the selected farms, were the same employed in the study of bovine brucellosis for Parana State in the context of National Program for Control and Eradication of Brucellosis and Tuberculosis. The sampling was performed in two stages. Blood samples were collected from 1930 females aged '> OU ='24 months from 295 non vaccinated herds. Serum samples were tested for antibodies against BoHV-1 using an indirect ELISA. The epidemiological questionnaire was applied on all the selected farms and aimed to obtain epidemiological data. Hundred ninety of the 295 herds were positive for BoHV-1, presenting prevalence of positive herds of 64.41% [58.65-69.87%]. The number ('> OU ='23) of females aged 24 months (OR=2.22; IC: 1.09-4.51), purchased cattle (OR=2.68; IC: 1.48-4.82), use of common grass (OR=5.93; IC: 1.31-26.82), history of abortion in the last 12 months (OR=2.37; IC: 1.09-5.16) and presence of wildlife animals (OR=8.86; IC: 1.11-70.73) were identified as risk factors for BoHV-1 infection in the multivariate logistic regression. These results indicate that BoHV-1 infection is widespread in the studied region and that factors related to the herd characteristic and management are associated with the infectio

Seroepidemiology Survey of Bovine Alphaherpesvirus 1 (BoHV-1) in Unvaccinated Beef Bubalines (Bubalus bubalis) from Southern Brazil

Background: The bovine alphaherpesvirus type 1 (BoHV-1) belonging to the order Herpesvirales, family Herpesviridae, subfamily Alphaherpesvirinae and genus Varicellovirus.Bovine alphaherpesvirus 1 (BoHV-1) infections have a great importance due to the high rates of dissemination in cattles worldwide. Although, the BoHV-1 was largely related in cattle, buffaloes were also classified as host for the virus. However, studies that determine seroepidemiological data in this species are scarce and necessary. The aim of the present study was to determine the presence of antibodies against BoHV-1 in healthy beef buffaloes using the virus neutralization (VN) technique.Cases: This work portrays an epidemiological survey, based on a sectional study characterized by blood samples collected from 54 Murrah buffalo, aged 6 to 24 months, from the municipalities of Guaraqueçaba, Ponta Grossa, Antonina and Doutor Ulysses, located in Paraná state, being 20, 14, 10 and 10 samples from each location, respectively. Thirty-seven percent (20/54) of the samples were collected at slaughterhouse with registration at the Federal Inspection Service (SIF) of the Ministry of Agriculture, Livestock and Food Supply (MAPA), and 63% (34/54) at the buffalo's farms. The serum samples were collected in sterile tubes without anticoagulant in stored isothermal boxes, with serum separation and stored at -20ºC. The samples were sent to the Animal Virology Laboratory of Universidade Estadual de Londrina, UEL for serological analysis. Serological diagnosis using the virus neutralization (VN) technique was performed according the OIE manual of diagnostic tests and vaccines for terrestrial animals, using the BoHV-1 Los Angeles strain. Forth-five of the 54 samples (83.3%) evaluated generated titers of antibodies against BoHV-1 present in all evaluated herds, and the samples collected in herd from Antonina city were 100% positive, following by 80% in Guaraqueçaba and Doutor Ulysses city, and 78.6% in Ponta Grossa city. In relationship to the titration of anti-bovine herpesvirus 1 antibodies (BoHV-1), it was found that 23/45 (51.12%) of the samples had titers ≤ 16, 13/45 (28.88 %) with indexes between 32-64, and 9/45 (20%) with levels above 128.Discussion: The confirmation of the presence of antibodies against BoHV-1 in the buffalo samples tested in the present study proves the circulation of the agent in the studied species, with a homogeneous distribution of bovine alphaherpesvirus 1 in all the evaluated herds. There was high variability in antibody titers against BoHV-1 from all herds, and the occurrence of a high number of buffaloes reactive to BoHV-1 and with high antibody titers suggests a course of active or reactive infections in these animals. In only one of the herds, the age of the animals studied varied between 6 to 8 months. The fact that these animals are lactating raises the possibility of anti-BoHV-1 antibodies being acquired by breastfeeding. The results obtained allow to conclude, considering the fact that the animals are not vaccinated against BoHV-1, the presence of circulating antibodies is from a natural infection, and the positivity in all farms tested denotes the endemicity of the BoHV-1 infection in herds. To the best knowledge of the authors, this is the first study on the detection and measurement of antibodies against BoHV-1 in healthy and unvaccinated buffaloes in the state of Paraná, southern Brazil, showing a high frequency of seroreagent animals.

Ultrastructure of Sheep Primordial Follicles Cultured in the Presence of Indol Acetic Acid, EGF, and FSH

The aim of this study was to investigate the ultrastructural characteristics of primordial follicles after culturing of sheep ovarian cortical slices in the presence of indol acetic acid (IAA), Epidermal Growth Factor (EGF), and FSH. To evaluate ultrastructure of primordial follicles cultured in MEM (control) or in MEM containing IAA, EGF, and FSH, fragments of cultured tissue were processes for transmission electron microscopy. Except in the control, primordial follicles cultured in supplemented media for 6 d were ultrastructurally normal. They had oocyte with intact nucleus and the cytoplasm contained heterogeneous-sized lipid droplets and numerous round or elongated mitochondria with intact parallel cristae were observed. Rough endoplasmic reticulum (RER) was rarely found. The granulosa cells cytoplasm contained a great number of mitochondria and abundant RER. In conclusion, the presence of IAA, EGF, and FSH helped to maintain ultrastructural integrity of sheep primordial follicles cultured in vitro

Serological and molecular investigation of viral agents in free-living jaguars of the Pantanal wetlands, state of Mato Grosso, Brazil

Este estudo investigou a exposição de onças-pintadas de vida livre a agentes virais selecionados em duas unidades de conservação federais no Pantanal de Mato Grosso, Brasil. Para a análise desses agentes virais, a maioria de caráter zoonótico, foram utilizados métodos sorológicos e moleculares. Nenhuma das onze onças-pintadas examinadas foi positiva na técnica de real-time RT-PCR para a detecção molecular dos agentes da Influenza aviária e Febre do Nilo Ocidental (WNF). Somente um animal foi positivo sorologicamente para a o vírus da Encefalite Equina do Leste (EEE) pela Microtécnica de vírus neutralização em culturas de células VERO, sendo este o primeiro relato da exposição de onças-pintadas. Todos os animais examinados s foram negativos para o vírus da Encefalite Equina do Oeste (WEE) e Venezuelana (VEE). Amostras de soro colhidas de 11 onças-pintadas foram submetidas a adois testes distintos para a detecção de anticorpos contra o vírus da raiva (Teste Rápido de Inibição de Foco de Fluorescência – RFFIT e Microteste Simplificado de Inibição da Fluorescência - SFIMT), resultando em cinco animais positivos, dos quase dois positivos para cada teste e um positivo quando submetido aos dois testes sorológicos. Além disso, três das 14 amostras submetidas a técnica de soroneutralização foram positivas para a pesquisa de anticorpos contra o vírus da cinomose (CDV) e 15 amostras positivas das 17 analisadas para a pesquisa de anticorpos contra o parvovírus canino (CPV) foram identificadas pela técnica de Inibição da Hemaglutinação (HI). De acordo com os resultados deste estudo, é pouco provável que os agentes virais aqui analisados representem ameaça à população de onças-pintadas nesta região.This study investigates the exposure of free-living jaguars from two federal protected areas in the Pantanal of Mato Grosso, Brazil, to a variety viral agents. These viral agents, particularly causing zoonotic diseases, were analyzed using serological and molecular methods. None of the jaguars was positive by RT-PCR for the molecular detection of avian influenza and West Nile Fever (WNF). Only one animal was serologically positive for Eastern Equine Encephalitis (EEE) by virus neutralization test in VERO cell cultures, representing the first reported case of jaguar exposure to EEE virus. However, all the animals were negative for Western Equine Encephalitis (WEE) virus and Venezuelan Equine Encephalitis (VEE) virus. Eleven jaguars were tested by two tests for the detection of antibodies against rabies virus (Simplified Fluorescent Inhibition Microtest – SFIMT and Rapid Fluorescent Focus Inhibition Test – RFFIT), resulting in five positive animals, two animals in each test and one in both serological tests. Furthermore, three out of 14 samples subjected to the neutralization test were positive for antibodies against canine distemper virus (CDV), and 15 out of 17 samples subjected to the hemagglutination-inhibition test (HI) were positive for antibodies against canine parvovirus (CPV). In view of the findings of this study, it is unlikely that the viruses examined here represent a threat to the jaguar populations in this region

First report of infectious spleen and kidney necrosis virus (ISKNV) in two native cichlids cultured in Brazil

Peacock bass (syn.: tucunaré, Cichla ocellaris) and the pearl cichlids (syn.: acará, Geophagus brasiliensis) are South American cichlids that are highly valued in both the ornamental and sport fish industries. Since 2017, a number of outbreaks of infectious spleen and kidney necrosis virus (ISKNV) have been reported on Brazilian food and ornamental fish farms. In this study, we detected ISKNV in farmed peacock bass and pearl cichlid by PCR and sequence analysis of the partial major capsid protein (MCP) gene. Moribund peacock bass (n=10) and pearl cichlids (2) from a farm experiencing elevated mortality among juveniles and adults of these species, were submitted for bacteriological and molecular diagnostics. Spleen, liver, brain, and kidney tissues were cultured on 5% sheep blood agar and cystine heart agar with 1% glucose and bovine haemoglobin. No bacteria were isolated from the 12 fish. Additionally, DNA extracts from the liver and spleen of all animals were tested for ISKNV using two conventional polymerase chain reaction (cPCR) assays and two nested PCR (nPCR) assays. ISKNV DNA was amplified in all 12 fish DNA extracts tested, in two or more of the PCR assays. Selected ISKNV amplicons were confirmed by Sanger sequencing. The nucleotide sequences derived from these animals were identical to ISKNV strains previously detected in food (e.g., tilapia and carp) and ornamental species, including strains previously detected in fish from Brazil. To the authors’ knowledge, this is the first report of ISKNV in these native Brazilian cichlids