Reports of bovine listeriosis in Brazil are uncommon, being restricted to citations within retrospective studies, resulting in scarce documented information of this important disease of cattle. This manuscript describes the molecular findings associated with spontaneous encephalitic listeriosis in two steers from distinct herds within the state of Paraná, southern Brazil. Both animals demonstrated altered consciousness suggestive of brain stem dysfunctions and died a few days after the initial onset of disease. Polymerase chain reaction (PCR) assays were designed to target specific genes of infectious neurological agents of cattle. These included bovine herpesvirus 1 and 5 (BoHV-1 and BoHV-5), ovine herpesvirus 2 (OvHV-2), Listeria monocytogenes, and Histophilus somni. Rabies virus was discarded in evaluations done at the official state diagnostic laboratory. Gross alterations were insignificant; histopathology demonstrated rhombencephalitis associated with macrophage-predominant, multifocal to coalescing microabscesses and extensive perivascular cuffings in both steers. The L. monocytogenes PCR assay amplified the 172-bp amplicon of the listeriolysin gene from the brain stem of both animals and from the telencephalon, thalamus, and cerebellum of one of them. Phylogenetic analyses demonstrated that the strains derived from this study clustered with known strains of L. monocytogenes lineage I. The BoHV-1 and BoHV-5, OvHV-2, and H. somni PCR assays were negative. These results confirm the participation of L. monocytogenes lineage I in the etiopathogenesis of the neurological disease herein described and represent the first complete description of encephalitic listeriosis in cattle from Brazil.
RESUMOO objetivo deste trabalho foi determinar a prevalência das infecções latentes por BoHV-1 e por BoHV-5 em bovinos de corte criados no Estado do Paraná. Os gânglios do nervo trigêmeo foram coletados de 400 bovinos hígidos, entre 18 e 36 meses de idade, provenientes de 90 propriedades rurais localizadas em diferentes mesorregiões geográficas do Estado e abatidos em frigorífico com Serviço de Inspeção Federal. A reação em cadeia da polimerase com amplificação do gene que codifica a glicoproteína C foi empregada para a detecção do DNA viral. Cento e nove bovinos eram herpéticos (27,25%), sendo 14,25% (57/400) infectados com BoHV-1, 9,75% (39/400) infectados com BoHV-5 e 3,25% (13/400) portadores de infecção mista. A distribuição geográfica foi heterogênea e as infecções foram mais prevalentes nas mesorregiões localizadas ao norte do Estado. A vigilância para a encefalite por BoHV-5 deve ser intensificada na mesorregião Noroeste. Palavras-chave: herpes-vírus bovino, diagnóstico, epidemiologia, doenças neurológicas ABSTRACT The prevalence of latent infection with BoHV-1 or
Presença do genoma de BoHV-5 no líquido cefalorraquidiano de bovinos com meningoencefalite herpética (Isernhagen et al., 2011). A evolução até a morte é variável, podendo ser de um a 15 dias (Elias et al., 2004;Isernhagen et al., 2011).Diferentes técnicas podem ser utilizadas para a confirmação do diagnóstico post mortem, como histopatologia, imuno-histoquímica, isolamento viral e a reação em cadeia da polimerase (PCR). As coletas do LCR foram realizadas no espaço atlanto-occipital, utilizando-se a agulha metálica do cateter intravenoso 18G com 5cm de comprimento (BD Angiocath) e aspiração com seringa de 1mL. O LCR foi mantido refrigerado até a chegada ao laboratório. A eutanásia, seguida de necropsia, foi realizada após a coleta do LCR, com a finalidade de confirmação do diagnóstico e em comum acordo com o proprietário, considerando a evolução da doença, a gravidade das manifestações clínicas e a probabilidade reduzida de resposta a tentativas de tratamentos. Fragmentos do encéfalo foram http://dx
The clinical, pathological, and molecular findings due to malignant catarrhal fever lesions induced by ovine herpesvirus type 2 (OvHV-2) are described in a 12-month-old, mixed-breed, heifer from northern Paraná, Brazil. The animal demonstrated clinical signs including manifestations of aggressive behavior, nystagmus, incoordination, decumbency, paddling movements, and death within 48 hours, after which was submitted for routine necropsy. Brain fragments were used in PCR assays for identification of infectious agents (OvHV-2; bovine herpesvirus-1 and -5; and Histophilus somni). Rabies was discarded in text done at the official state diagnostic laboratory. Significant pathological alterations include disseminated vasculitis with lymphoplasmacytic inflammatory infiltrate within the brain, kidneys, liver, and myocardium. PCR assay amplified the specific 422 base pair fragment of the OvHV-2 tegument protein gene from the brain; all other evaluations were negative. These findings confirm the participation of OvHV-2 in the etiopathogenesis of the disease observed in this mixed-breed heifer.
RESUMO: O objetivo deste trabalho foi identificar as doenças neurológicas que acometeram bovinos no estado do Paraná entre os anos de 2009 e 2015. A investigação aconteceu, preferencialmente, nas propriedades rurais onde os casos ocorreram. Foram registradas as informações sobre a evolução das doenças nos bovinos afetados do rebanho, e os prováveis fatores de risco foram identificados. Todos os procedimentos de exame físico geral e neurológico foram realizados sistematicamente para a caracterização da síndrome neurológica presente. Amostras de sangue e de líquor foram colhidas para a realização de exames laboratoriais. De acordo com o tempo de evolução e com a gravidade dos sinais clínicos observados, os bovinos doentes eram mantidos vivos para acompanhamento da evolução ou da resposta ao tratamento, ou eram submetidos à eutanásia seguida de necropsia. Fragmentos do sistema nervoso e dos demais órgãos foram colhidos para exame histopatológico. O exame de imunofluorescência direta e a prova biológica em camundongos foram realizados em todos os bovinos que morreram, com a finalidade de confirmar ou descartar o diagnóstico de raiva. Métodos laboratoriais específicos das rotinas de virologia, bacteriologia e toxicologia foram empregados, como complementares, para o estabelecimento do diagnóstico diferencial. Foram investigados 236 bovinos com doença neurológica, sendo 85 casos de ocorrência individual e 151 casos distribuídos por surtos que ocorreram em 79 rebanhos. As encefalopatias (180/236; 76,2%) predominaram sobre as mielopatias (27/236; 11,4%). As doenças inflamatórias determinadas por infecções (98/236; 41,5%) e as doenças tóxicas (91/236; 38,6%) foram as principais, enquanto as causas degenerativas (10/236; 4,2%), metabólicas (9/236; 3,8%), físicas (9/236; 3,8%), neoplásicas (4/236; 1,7%), e os defeitos congênitos (1/236; 0,4%) ocorreram menos frequentemente. Os casos inconclusivos somaram 5,9% (14/236). A meningoencefalite por BoHV-5 e a raiva foram as doenças de frequência maior e podem ser consideradas as mais importantes. Dentre as causas tóxicas, as intoxicações por plantas se destacaram (63/91; 69,2%) e foram responsáveis por 26,6% de todos os casos. Destacaram-se ainda a polioencefalomalácia, a meningoencefalite trombótica por Histophilus somni e o botulismo. Essas informações contribuem para que os médicos veterinários adotem condutas mais efetivas de diagnóstico e de prevenção, e são valiosas para o sistema oficial de vigilância epidemiológica do estado.
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.
Leptospirosis is a zoonosis that can infect buffaloes and other mammalians, and it’s caused by Gram-negative bacteria of the genus Leptospira spp. The infection occurs through contact with urine or a contaminated environment with the mucous membranes of the animals, and the disease has two phases: leptospiremia and leptospiruria. The objective of this study was to determine the frequency of leptospirosis in buffaloes in the state of Paraná, Brazil, as well as the main serovars present in this area. A total of 276 blood samples were collected from buffaloes from 18 properties located in different cities and mesoregions of Paraná state. A microscopic agglutination test (MAT) was performed to detect the presence of anti-Leptospira spp. antibodies and to identify its serovars. The prevalence was 77.17% (213/276) of buffaloes seropositive for at least one serovar. The most frequent serovar was Icterohaemorrhagiae, followed by the Pomona serovar and serovar Hardjo. This study seems to be the first to determine the frequency of buffaloes with leptospirosis in the state of Paraná. Prevention and control measures are important to control the disease in herds.
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