Gastric disorders of cattle in western Rio Grande do Sul State, Brazil / Distúrbios gástricos de bovinos no Oeste do Rio Grande do Sul

A retrospective study of gastric disorders in autopsied cattle in the Western region of Rio Grande do Sul State, was performed. The exam reports of bovine necropsy of the Veterinary Pathology Laboratory, Unipampa, were analyzed in the period from 2010 to 2018. All cases in which death was primarily caused by disturbance in the gastric chambers were included. During the period evaluated, 141 cattle were necropsied. Of those, 25 had gastric disorders. Of those, 53% had alterations in the rumen, followed by abomasum (17%), involvement of two chambers (13%) and reticulum (9%). Most cases corresponded to beef cattle raised in an extensive system and most them for calf production and fattening with an average age of approximately three years. The cases occurred in farms of four different municipalities. Bullous bloat by excessive Trifolium repens ingestion was the gastric disturbance with the highest number of dead cattle observed in this study, especially in irrigated áreas of livestock farms. Cases such as lactic acidosis, ruminal alkalosis due to excessive urea ingestion and Baccharis coridifolia poisoning were also important gastric disturbances in necropsied cattle, associated especially with poor management and period of scarcity of good quality fodder. Cases of Clostridium perfringens infection were also observed in young cattle suggesting that it is an important infectious agent in the evaluated cattle herds, also showing failures in vaccination of the herds. As observed, gastric disturbances in cattle in the western region of Rio Grande do Sul have several causes. Metabolic/toxic and infectious disturbances were important causes of mortality in the herds, inducing considerable economic losses. Based on this study, it is clear that the majority of outbreaks or isolated cases occurred due to errors in the management of the properties and the vast majority of them could have been avoided with improvements in the technical qualification of the workers and simple adjustments in the farming methods. It is also emphasized the importance of the conclusive diagnosis to control these disorders, once after the orientation to the producers, was observed significant decrease in cattle losses in the farms.(AU)

Foi realizado estudo retrospectivo dos distúrbios gástricos em bovinos necropsiados na região Oeste do Rio Grande do Sul. Foram analisados os relatórios de exame de necropsia de bovinos do Laboratório de Patologia Veterinária (LPV) da Universidade Federal do Pampa (Unipampa), Rio Grande do Sul, no período de 2010 a 2018. Foram incluídos todos os casos nos quais a morte foi causada primariamente pelo distúrbio nas câmaras gástricas. De um total de 141 bovinos necropsiados, 25 corresponderam a distúrbios gástricos. Dentre esses, 53% apresentaram alterações no rúmen, seguido de abomaso 17%, acometimento concomitante de duas câmaras 13% e retículo 9%. A maioria dos casos ocorreram em bovinos de corte criados em sistema extensivo e a maioria destinados à produção de bezerros e engorda com média de idade de aproximadamente três anos. Os casos ocorreram em propriedades rurais de quatro municípios da região Oeste do estado. O timpanismo bolhoso por ingestão excessiva de Trifolium repens foi o distúrbio gástrico com maior número de bovinos mortos observados nesse estudo, especialmente em propriedades com criação de animais em áreas de irrigação. Casos como acidose láctica, alcalose ruminal por intoxicação por ureia e intoxicação por Baccharis coridifolia também foram importantes distúrbios gástricos nos bovinos necropsiados e percebeu-se sua associação a falhas no manejo e à época de escassez de forragem de boa qualidade. Foram observados ainda casos de infecção por Clostridium perfringens em bovinos jovens o que sugere também tratar-se de um importante agente infeccioso nos rebanhos bovinos avaliados, demonstrando ainda falhas na vacinação dos rebanhos. Conforme observado, diversos são os distúrbios gástricos em bovinos na região Oeste do Rio Grande do Sul, tendo como importantes causas de mortalidades os distúrbios metabólicos/tóxicos e infecciosos, induzindo consideráveis perdas econômicas. Com base nesse levantamento, percebe-se que a maioria dos surtos ou casos isolados estudados ocorreram por erros no manejo nas propriedades e, na sua grande maioria, poderiam ter sido evitados com especialização da mão de obra e ajustes simples. Ressalta-se ainda a importância do diagnóstico conclusivo para controle desses distúrbios, uma vez que, após a orientação aos produtores, observou-se significativa diminuição das perdas de bovinos nas propriedades.(AU)

Clostridial diseases diagnosed in cattle from the South of Rio Grande do Sul, Brazil. A forty-year survey (1978-2018) and a brief review of the literature / Clostridioses diagnosticadas em bovinos no sul do Rio Grande do Sul. Uma pesquisa de quarenta anos (1978-2018) e breve revisão da literatura

Clostridial diseases are important causes of livestock losses in the southern Rio Grande do Sul. Since 1978 annual surveys conducted at the "Laboratório Regional de Diagnóstico" of the "Universidade Federal de Pelotas" (LRD-UFPel) have shown that clostridial diseases represent 10.40% of the bacterial diseases diagnosed in cattle and 1.65% of all diseases diagnosis in cattle over a 40-year period. The purpose of this study is to review the clinical, epidemiological and pathological aspects of the clostridial diseases diagnosed in cattle from January 1978 to December 2018 at the LRD-UFPel in the hopes that it will constitute a useful guide for field veterinary practitioners and interested farmers. We assessed and review the necropsy protocols of 6,736 cattle; these necropsies were performed either by LRD-UFPel faculty or by field veterinary practitioners; 111 outbreaks (1.65%) were diagnosed as clostridial disease, distributed as follows: 35 outbreaks of tetanus, 34 of blackleg, 23 of bacillary hemoglobinuria, 11 of malignant edema (gas gangrene), and eight of botulism. Approximately 904, from a total of 42,480 cattle at risk, died in these outbreaks.(AU)

Clostridioses são doenças produzidas por alguma das espécies do gênero Clostridium e são importantes causas de perdas pecuárias no sul do Rio Grande do Sul. Pesquisas anuais realizadas no Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas (LRD-UFPel) desde 1978 demonstraram que as clostridioses representaram 11,1% das doenças bacterianas diagnosticadas em bovinos e 1,65% de todos os diagnósticos de doenças em bovinos ao longo de 40 anos. O objetivo deste estudo é revisar os aspectos clínicos, epidemiológicos e patológicos das clostridioses diagnosticadas de janeiro de 1978 a dezembro de 2018, pelo LRD/UFPel com a intenção de que esse trabalho possa servir de guia útil para os veterinários de campo e fazendeiros interessados. Foram avaliados e revisados os protocolos de necropsia de 6.736 bovinos; essas necropsias foram realizadas pelo pessoal do LRD/UFPel ou por veterinários de campo. Cento e quatro (1,16%) casos foram diagnosticados como clostridioses, distribuídos da seguinte forma: 35 surtos de tétano, 34 de carbúnculo sintomático, 23 de hemoglobinúria bacilar, 11 de edema maligno (gangrena gasosa) e oito de botulismo. Aproximadamente 904, de um total de 42.480 bovinos sob-risco, morreram nesses surtos.(AU)

Blackleg in a free-range brown brocket deer (Mazama gouazoubira) / Carbúnculo sintomático em veado virá de vida livre (Mazama gouazoubira)

A case of blackleg in a brown brocket deer (Mazama gouazoubira) associated with trauma from being hit by a car in southern Rio Grande do Sul is reported. The clinical signs included fever, dehydration and lethargy that worsened progressively until 36 hours after the accident, when the animal died. In the fore right limb, there was a comminuted closed fracture of the radius and ulna but no skin wounds were observed. Grossly, the musculature of the pelvic limbs presented hemorrhage, edema and emphysema. Microscopically, the muscles of both rear legs had necrosis, edema, hemorrhage and mild inflammatory infiltration of neutrophils. Clostridium chauvoei was cultured from affected skeletal muscles, and it was also detected by immunohistochemistry, confirming a diagnosis of blackleg. The overlapping habitat of cattle and brown brocket deer is proposed as a predisposing factor in this case and alerts to spillover cases maybe happening in this region. In addition, blackleg should be included as differential diagnoses of deer with post-traumatic myositis.(AU)

Descreve-se um caso de carbúnculo sintomático em um veado-virá (Mazama gouazoubira), macho, jovem, resgatado após atropelamento em uma rodovia na região sul do Rio Grande do Sul. O cervídeo apresentou febre, desidratação e letargia, progredindo para a morte em 36 horas. No membro torácico direito foi observado fratura cominutiva fechada de rádio e ulna sem a presença de feridas perfurantes. Na necropsia foi observada hemorragia, edema e enfisema na musculatura dos membros pélvicos. Microscopicamente, os músculos dos membros pélvicos apresentaram necrose, edema, hemorragia e discreto infiltrado inflamatório neutrofílico. Houve o isolamento de Clostridium chauvoei e marcação positiva na técnica de IHQ com anticorpo monoclonal anti-C. chauvoei, confirmando o diagnóstico de carbúnculo sintomático. A sobreposição de habitat entre bovinos domésticos e cervídeos pode ser um fator de risco para esta doença e chama a atenção para casos de "spillover" que podem estar ocorrendo na região. Adicionalmente, sugere-se que o carbúnculo sintomático seja incluído nos diagnósticos diferenciais de cervídeos que apresentam miosite pós-traumática.(AU)

Adhesion and invasion of Clostridium perfringens type A into epithelial cells

ABSTRACT Clostridium perfringens is the causative agent for necrotic enteritis. It secretes the major virulence factors, and α- and NetB-toxins that are responsible for intestinal lesions. The TpeL toxin affects cell morphology by producing myonecrosis, but its role in the pathogenesis of necrotic enteritis is unclear. In this study, the presence of netB and tpeL genes in C. perfringens type A strains isolated from chickens with necrotic enteritis, their cytotoxic effects and role in adhesion and invasion of epithelial cells were evaluated. Six (27.3%) of the 22 C. perfringens type A strains were harboring the tpeL gene and produced morphological alterations in Vero cells after 6 h of incubation. Strains tpeL (-) induced strong cell rounding after 6 h of incubation and produced cell enlargement. None of the 22 strains harbored netB gene. All the six tpeL (+) gene strains were able to adhere to HEp-2 cells; however, only four of them (66.6%) were invasive. Thus, these results suggest that the presence of tpeL gene or TpeL toxin might be required for the adherence of bacteria to HEp-2 cells; however, it could not have any role in the invasion process.

Producción a nivel de laboratorio de Clostridium septicum IRP15 para la formulación de una vacuna veterinaria / Laboratory production of Clostridium septicum IRP15 for the formulation of a veterinary vaccine

Objetivo: establecer las condiciones de producción a nivel de laboratorio de la alfa toxina de Clostridium septicum IRP15 para la formulación de una vacuna veterinaria y la optimización del proceso de producción. Métodos: se caracterizó y estandarizó la edad apropiada del inóculo para los cultivos en un fermentador New Brunswick Scientific 7 L. Las condiciones de cultivo fueron: cepa C. septicum IRP15, medio de cultivo VBH, 5 L/vaso de 7 L, inóculo de 250 mL (5 por ciento), 37 ºC, 24 h, bajo agitaciones prueba de 0, 25 y 50 r.p.m. Se estableció el perfil cinético morfológico, de biomasa, consumo de sustrato y producción de toxina. Resultados: para las fermentaciones de 0 y 25 r.p.m. no se presentó fase de adaptación; el microorganismo creció de manera exponencial hasta las 4 y 6 h de fermentación, consumiendo simultáneamente la mayor cantidad de glucosa presente en el medio. A partir de estas horas y hasta las 24, se realizó la prueba de DL50 en ratones y se destaca que a 25 r.p.m. se obtuvo el mayor título de toxina (1/23). En las fermentaciones a 50 r.p.m. se observó que el microorganismo experimenta una fase de adaptación de 4 h aproximadamente; con un retardo en producción de biomasa, consumo de glucosa y producción de toxina, condición que no resulta óptima para la producción del antígeno. Conclusiones: la producción de toxina se presenta en la fase logarítmica y durante la fase estacionaria, asociándose así al crecimiento y al fenómeno de esporulación(AU)

Objective: to set the laboratory production conditions of Clostridium septicum IRP15 alpha toxin for the formulation of a veterinary vaccine and the optimization of the productive process. Methods: the appropriate inoculum age for the cultures was characterized and standardized in a 7L New Brunswick Scientific biorreactor. The conditions of culturing were C. septicum IRP15 strain, VBH medium at 5 L/7 L glass, 250 mL (5 percent) inoculum, 37 ºC, and 24 h under shaking conditions of 0, 25 y 50 r.p.m. The following kinetic parameters were monitored: morphological changes, biomass production, glucose consumption and toxin production. Results: for the shaking conditions at 0 and 25 r.p.m., C. septicum did not show an adaptation phase growth. The bacteria kept growing at the log phase up to 4-6 hours of fermentation respectively, thus consuming the highest amount of glucose from the medium. As from the growth phase hours till the 24 h of cultivation, the 50 percent lethal dose (LD50) in mice assay was conducted and at 25 r.p.m. condition, the best titre of toxin was reached (1/23). The cultures at 50 r.p.m. condition showed that the bacteria experienced adaptation phase for almost four hours, resulting in delayed biomass production, glucose consumption and toxin production. These results suggested that 50 r.p.m. is not useful for the antigen production. Conclusions : the toxin production occurred at the log phase and during the stationary phase, thus it is associated to growth and to sporulation(AU)

Acute myonecrosis in horse caused by Clostridium novyi type A

The objective of this study was to describe the first report involving a case of equine acute myonecrosis caused by C. novyi type A with an emphasis on clinical signs, the pathological and bacteriological analysis, and molecular identification of the microorganisms as the key of the definitive diagnosis.

Characterization of Clostridium perfringens isolated from mammals and birds from Guwahati city, India

Of the 102 samples collected from mammals and birds, both domestic and captive wild, 48 were found to be positive for Clostridium perfringens. Most of the mammal isolates (84.38%) appeared to have been collected from clinically affected animals, while 33.33% of the bird samples were from clinically affected and 21.43% from apparently healthy birds infected with C. perfringens. Isolates revealed high sensitivity to ciprofloxacin, enrofloxacin and norfloxacin. Among the isolated C. perfringens, 30 (62.50%) showed DNase production. Hemolytic activity was recorded in 14 (24.16%) of the isolates and 28 (58.33%) showed phospholipase C production. All the phospholipase C positive isolates revealed the presence of cpa gene encoding alpha (α) toxin. Of the 102 samples collected from mammals and birds, both domestic and captive wild, 48 were found to be positive for Clostridium perfringens. Most of the mammal isolates (84.38%) appeared to have been collected from clinically affected animals, while 33.33% of the bird samples were from clinically affected and 21.43% from apparently healthy birds infected with C. perfringens. Isolates revealed high sensitivity to ciprofloxacin, enrofloxacin and norfloxacin. Among the isolated C. perfringens, 30 (62.50%) showed DNase production. Hemolytic activity was recorded in 14 (24.16%) of the isolates and 28 (58.33%) showed phospholipase C production. All the phospholipase C positive isolates revealed the presence of cpa gene encoding α toxin.(AU)

Toxinas de Clostridium perfringens / Toxins of Clostridium perfringens

Clostridium perfringens es un bacilo grampositivo anaerobio con capacidad de formar esporas. Es uno de los patógenos bacterianos con mayor distribución en el medio ambiente, ya que puede ser aislado de muestras de suelo y de agua y además forma parte de la microbiota intestinal de animales y humanos. Sin embargo, en ciertas ocasiones puede actuar como patógeno oportunista y causar enfermedades como la gangrena gaseosa, la enterotoxemia del ovino y del caprino y la disentería del cordero, entre otras. En humanos, está asociado a enfermedades como la intoxicación por alimentos, la enterocolitis necrotizante en niños y la enteritis necrótica o pigbel de las tribus de Papúa-Nueva Guinea. El renovado interés que existe actualmente en el estudio de C. perfringens como patógeno veterinario y humano, junto con el avance de la biología molecular, han hecho posible que la ciencia tenga hoy un conocimiento más profundo sobre la biología y la patogenia de esta bacteria. En esta revisión bibliográfica se discuten y actualizan los principales aspectos de la patogenia intestinal de C. perfringens teniendo en cuenta las toxinas con mayor importancia médica descritas hasta el presente.

Clostridium perfringens is an anaerobic gram-positive spore-forming bacillus. It is one of the pathogens with larger distribution in the environment; it can be isolated from soil and water samples, which also belongs to the intestinal flora of animals and humans. However, on some occasions it can act as an opportunistic pathogen, causing diseases such as gas gangrene, enterotoxemia in sheep and goats and lamb dysentery, among others. In human beings, it is associated to diseases such as food poisoning, necrotic enterocolitis of the infant and necrotic enteritis or pigbel in Papua-New Guinea tribes. The renewed interest existing nowadays in the study of C. perfringens as a veterinarian and human pathogen, together with the advance of molecular biology, had enabled science to have deeper knowledge of the biology and pathology of these bacteria. In this review, we discuss and update the principal aspects of C. perfringens intestinal pathology, in terms of the toxins with major medical relevance at present.

Detection of several clostridia by a direct fluorescent antibody test in formalin-fixed, paraffin-embedded tissues

Descreve-se a aplicabilidade de uma técnica de imunofluorescência direta, para o diagnóstico de mionecroses causadas por clostrídios, a partir de tecidos fixados em formol e incluídos em parafina. Essa técnica pode auxiliar no diagnóstico do carbúnculo sintomático e da gangrena gasosa, contribuindo para determinar a real prevalência dessas doenças no país

Susceptibility of brown Swiss cattle to black leg disease

In a mixed herd of brown swiss and fresian cows, 7 fatal cases of black leg were recorded in brown swiss cows kept under same conditions with fresian cows. Clostridium chauvoei was the only organism increminated in these cases. Preventive measures including treatment and vaccination were successful to prevent further appearance of the disease

An outbreak of type C botulism in broiler chickens in Säo Paulo State, Brazil

Um surto por botulismo tipo C acometeu 12.000 frangos de sete semanas de idade no Estado de Säo Paulo. A fonte direta de toxina näo foi detectada. Toxina de Clostridium botulinum tipo C foi demonstrada no soro de aves vivas e nos extratos do conteúdo do "papo", moela e intestinos de aves com sintomas de botulismo. Estes extratos provocaram toxemia letal para camundongos em 12 horas. A toxina foi especificamente neutralizada pela antitoxina de C. botulinum tipo C e inativada pelo aquecimento a 100-C por 10 minutos. Depois da retirada das carcaças e a desinfecçäo dos galpöes näo ocorreram mais mortes. Toxina botulínica do tipo C é considerada como principal suspeita das mortes