Whole genome sequencing of Bacillus anthracis isolated from animal in the 1960s, Brazil, belonging to the South America subclade.

Bacillus anthracis causes anthrax disease and can affect humans and other animals. This zoonotic disease has an impact on the economic and health aspects. B. anthracis population is divided into three major clades: A (with worldwide distribution), B, and C (restricted to specific regions). Anthrax is most common in agricultural regions of central and southwestern Asia, sub-Saharan Africa, Southern and Eastern Europe, the Caribbean, and Central and South America. Here, we sequenced by short and long reads technologies to generate a hybrid assembly of a lineage of B. anthracis recovered from animal source in the 1960s in Brazil. Isolate identification was confirmed by phenotypic/biochemical tests and MALDI-TOF MS. Antimicrobial susceptibility was performed by in-house broth microdilution. B. anthracis IAL52 was susceptible to penicillin, amoxicillin, doxycycline, levofloxacin, and tetracycline but non-susceptible to ciprofloxacin. IAL52 was classified as sequence type ST2, clade A.Br.069 (V770 group). Sequencing lineages of B. anthracis, especially from underrepresented regions, can help determine the evolution of this critical zoonotic and virulent pathogen.

Carbúnculo sintomático em bovino: evolução clínica e terapêutica / Symptomatic blackleg in bovine: clinical and therapeutic evolution

As clostridioses são infecções comuns em rebanhos bovinos, causadas por microrganismos do gênero Clostridium e que afetam principalmente animais mais jovens. O objetivo deste estudo é relatar a evolução clínica e tratamento de um bovino acometido com carbúnculo sintomático. A paciente não apresentava histórico vacinal para clostridiose, e pertencia a um lote de animais em que já haviam ocorrido três mortes súbitas em curto período de tempo. Na avaliação física do animal em decúbito esternal, foram constatados sinais clínicos típicos de carbúnculo sintomático, edema e crepitação em membro pélvico e região escapular, além de claudicação quando colocado em estação. Também foi observado edema na região da cabeça, porém este sinal clínico não é frequentemente relatado em literatura científica. Mesmo com prognóstico desfavorável devido à manifestação grave da doença e grande parte das pesquisas mostrarem taxa de letalidade de quase 100%, optou-se pela tentativa de realizar as medidas terapêuticas necessárias. Foi realizado tratamento suporte e administração de fármacos, destacando-se a antibioticoterapia com penicilina. Além disso, também foi conduzido o procedimento cirúrgico por meio de confecção de incisões na pele e no subcutâneo do membro afetado (método ainda não descrito na literatura científica) com objetivo de reduzir a proliferação do Clostridium. Após 32 dias de tratamento, o animal obteve a cura clínica completa. As medidas terapêuticas foram suficientes para recuperar da saúde da paciente.

Clostridial diseases are common infections in cattle herds, caused by microorganisms of the genus Clostridium that mainly affect younger animals. This study aims to report the clinical evolution and treatment of a bovine affected with symptomatic blackleg. The patient had no vaccination history for clostridial disease and belonged to a group of animals in which three sudden deaths had already occurred in a short time. In the physical evaluation of the animal in sternal recumbency, typical clinical signs of blackleg, edema and crackling in the pelvic limb and scapular region and lameness when placed in the station were determined. Edema was also observed in the head region, but this clinical sign is not frequently reported in the scientific literature. Even with an unfavorable prognosis due to the severe manifestation of the disease and a large part of the studies showing a lethality rate of almost 100%, it was decided to try to carry out the necessary therapeutic methods. Support treatment and drug administration were performed, especially antibiotic therapy with penicillin. In addition, a surgical procedure was carried out by making incisions in the skin and subcutaneous tissue of the affected limb (a method not yet described in the scientific literature) to reduce the proliferation of Clostridium. After thirty-two days of treatment, the animal got a complete clinical cure. Therapeutic measures were sufficient to recover the patient's health.

Seguridad ocupacional contra Bacillus anthracis en la industria de curtiembre peruana / Occupational safety against Bacillus anthracis in the Peruvian tannery industry

El carbunco es una zoonosis que afecta a animales herbívoros, causada por el microorganismo Bacillus anthracis; que se encuentra en su forma vegetativa en el organismo infectado, y una vez en contacto con el aire forma esporas que infectan al huésped, liberando toxinas que inducen edema, septicemia y necrosis tisular. En humanos, la transmisión se produce por cortes, pinchazos o contacto directo de piel lesionada con esporas o tejidos de animales infectados. En Perú, para la década de 1980, se reportaron 400 casos anuales en áreas rurales agrícolas, adquiridos por manipulación del ganado vacuno, muerto por ántrax. Es por ello, que la piel y el cuero procedente de animales infectados representa un riesgo de infección para los trabajadores de industrias curtiembres. Se realizó un estudio transversal para evaluar el riesgo biológico ocupacional y las medidas de bioseguridad relacionadas con el Bacillus anthracis en pequeñas, medianas y grandes empresas curtiembres peruanas. Se aplicó un instrumento a los trabajadores para cuantificar las variables, componentes de riesgo biológico y conocer las medidas higiénicas adoptadas. Posteriormente se calculó e interpretó el nivel de riesgo biológico. Se encontró que el nivel de riesgo supera el límite de exposición biológica para el Bacillus anthracis en todas las empresas evaluadas y los trabajadores de las pequeñas empresas tienen poco conocimiento acerca del riesgo biológico al que están expuestos, siendo necesaria la capacitación de los trabajadores acerca del Bacillus anthracis y adopción de medidas de bioseguridad en empresas curtiembres, que garanticen la salud de los trabajadores(AU)

Anthrax is a zoonosis that affects herbivorous animals, caused by the microorganism Bacillus anthracis; which is found in its vegetative form in the infected organism, and once in contact with the air it forms spores that infect the host, releasing toxins that induce edema, septicemia and tissue necrosis. In humans, transmission occurs through cuts, punctures, or direct contact of broken skin with spores or tissues of infected animals. In Peru, for the 1980s, 400 annual cases were reported in rural agricultural areas, acquired by handling cattle, which died of anthrax. For this reason, the skin and leather from infected animals represent a risk of infection for workers in tanneries. A cross-sectional study was carried out to evaluate the occupational biological risk and biosafety measures related to Bacillus anthracis in small, medium and large Peruvian tanneries. An instrument was applied to the workers to quantify the variables, components of biological risk and to know the hygienic measures adopted. Subsequently, the level of biological risk was calculated and interpreted. It was found that the level of risk exceeds the biological exposure limit for Bacillus anthracis in all the companies evaluated and the workers of the small companies have little knowledge about the biological risk to which they are exposed, being necessary the training of the workers about the Bacillus anthracis and adoption of biosafety measures in tanneries, which guarantee the health of workers(AU)

Can cats cause colossal contagious cutaneous carbuncles?

Sporothrix spp. cause the most common deep fungal skin infections in Brazil and this is related to infected cats. Transmission is traditionally from organic material/plants but can also be zoonotic. Culture of a skin biopsy is the golden standard for determination. Treatment with oral itraconazole approaches up to 95% efficacy in patients with cutaneous sporotrichosis.

Diagnosis of blackleg from cattle tissue impregnated in common filter paper / Diagnóstico de carbúnculo sintomático a partir de papel filtro comum impregnado com tecido bovino

Blackleg, an acute myonecrosis caused by Clostridium chauvoei, is usually underdiagnosed since the rapid transport of adequate samples for laboratory testing is difficult. This study tested a direct polymerase chain reaction (PCR) technique using common filter paper impregnated with cattle tissue samples obtained from animals suspected with blackleg. Twenty-five samples, belonging to eleven animals from Rio Grande do Sul State, Brazil, were analyzed. The direct PCR technique identified eight positive animals corroborating with results from microbiological culture. Skeletal muscle was the most common tissue type used in this study and when the animal was positive the pathogen was always detected in this tissue. Storage time of the impregnated filter paper at room temperature did not prove to be a limiting factor for the quality of the results indicating that this procedure can be carried out in the field and samples be sent in regular mail. Our results suggested that direct PCR of common filter paper impregnated with cattle tissue is a practical and economical alternative for the diagnosis of blackleg.

Carbúnculo sintomático, uma mionecrose aguda causada por Clostridium chauvoei, costuma ser subdiagnosticada, pois o transporte rápido de amostras adequadas para exames laboratoriais é complicado. O objetivo deste estudo foi testar a técnica de reação em cadeia da polimerase (PCR) direta, utilizando papel filtro comum impregnado com amostras de tecido bovino obtidas de animais suspeitos de carbúnculo sintomático. Foram analisadas 25 amostras, pertencentes a onze animais do estado do Rio Grande do Sul, Brasil. A técnica de PCR direta identificou oito animais positivos, corroborando com os resultados da cultura microbiológica. O músculo esquelético foi o tecido mais utilizado neste estudo e quando o animal foi positivo, o patógeno sempre foi detectado neste tecido. O tempo de armazenamento do papel filtro impregnado, à temperatura ambiente, não se mostrou um fator limitante para a qualidade dos resultados, indicando que esse procedimento pode ser realizado no local e as amostras enviadas por correio normal. Nossos resultados sugerem que a PCR direta usando papel filtro comum impregnado com tecido bovino é uma alternativa prática e econômica para o diagnóstico de carbúnculo sintomático.

Diagnosis of blackleg from cattle tissue impregnated in common filter paper / Diagnóstico de carbúnculo sintomático a partir de papel filtro comum impregnado com tecido bovino

Blackleg, an acute myonecrosis caused by Clostridium chauvoei, is usually underdiagnosed since the rapid transport of adequate samples for laboratory testing is difficult. This study tested a direct polymerase chain reaction (PCR) technique using common filter paper impregnated with cattle tissue samples obtained from animals suspected with blackleg. Twenty-five samples, belonging to eleven animals from Rio Grande do Sul State, Brazil, were analyzed. The direct PCR technique identified eight positive animals corroborating with results from microbiological culture. Skeletal muscle was the most common tissue type used in this study and when the animal was positive the pathogen was always detected in this tissue. Storage time of the impregnated filter paper at room temperature did not prove to be a limiting factor for the quality of the results indicating that this procedure can be carried out in the field and samples be sent in regular mail. Our results suggested that direct PCR of common filter paper impregnated with cattle tissue is a practical and economical alternative for the diagnosis of blackleg.(AU)

Carbúnculo sintomático, uma mionecrose aguda causada por Clostridium chauvoei, costuma ser subdiagnosticada, pois o transporte rápido de amostras adequadas para exames laboratoriais é complicado. O objetivo deste estudo foi testar a técnica de reação em cadeia da polimerase (PCR) direta, utilizando papel filtro comum impregnado com amostras de tecido bovino obtidas de animais suspeitos de carbúnculo sintomático. Foram analisadas 25 amostras, pertencentes a onze animais do estado do Rio Grande do Sul, Brasil. A técnica de PCR direta identificou oito animais positivos, corroborando com os resultados da cultura microbiológica. O músculo esquelético foi o tecido mais utilizado neste estudo e quando o animal foi positivo, o patógeno sempre foi detectado neste tecido. O tempo de armazenamento do papel filtro impregnado, à temperatura ambiente, não se mostrou um fator limitante para a qualidade dos resultados, indicando que esse procedimento pode ser realizado no local e as amostras enviadas por correio normal. Nossos resultados sugerem que a PCR direta usando papel filtro comum impregnado com tecido bovino é uma alternativa prática e econômica para o diagnóstico de carbúnculo sintomático.(AU)

Sporadic human cutaneous anthrax outbreak in Shaanxi Province, China: report of two cases from 2018

ABSTRACT China's compulsory annual livestock anthrax vaccination policy has remarkably reduced but not completely eradicated human anthrax infections. Herein we describe a sporadic human cutaneous anthrax outbreak involving two cases in 2018 in Shaanxi Province, both involving herdsman who dealt with unvaccinated and potentially sick cattle. Both patients showed Bacillus anthracis-positive blister smear and blood culture. Treatment with penicillin was followed by uneventful recovery for both. The prompt performance of the prophylactic measures successfully interrupted the further transmission of this sporadic human cutaneous anthrax outbreak.

Sporadic human cutaneous anthrax outbreak in Shaanxi Province, China: report of two cases from 2018.

China's compulsory annual livestock anthrax vaccination policy has remarkably reduced but not completely eradicated human anthrax infections. Herein we describe a sporadic human cutaneous anthrax outbreak involving two cases in 2018 in Shaanxi Province, both involving herdsman who dealt with unvaccinated and potentially sick cattle. Both patients showed Bacillus anthracis-positive blister smear and blood culture. Treatment with penicillin was followed by uneventful recovery for both. The prompt performance of the prophylactic measures successfully interrupted the further transmission of this sporadic human cutaneous anthrax outbreak.

Bovine abortion by a vaccine strain of Bacillus anthracis / Aborto bovino por uma cepa vacinal de Bacillus anthracis

This paper reports the abortion of a male Aberdeen Angus bovine by a vaccine strain of Bacillus anthracis, describing the pathological and microbiological findings and the genome sequence. Necropsy findings included multifocal areas of hemorrhage in different organs. Histologically, various organs showed hemorrhage, fibrin exudation, necrosis associated with countless bacillary bacterial clumps and severe neutrophilic inflammatory infiltrate. In the microbiological examination, numerous rough, nonhemolytic, gray and dry colonies with irregular edges were isolated from liver, lung and abomasum content samples. Gram staining revealed square-ended Gram-positive rods arranged in chains. B. anthracis identification was confirmed by detection of the molecular chromosomal marker Ba813. The genomes from the isolated B. anthracis (named SPV842_15) and from the isolated vaccinal strain (Brazilian vaccinal strain), which was recovered from a commercial vaccine used in the pregnant cow, were sequenced. Genomic comparisons displayed a high level of nucleotide identity in the comparisons between B. anthracis SPV842_15 and the B. anthracis Brazilian vaccinal strain (98,2%). Furthermore, in both strains, only the plasmid pX01 sequence was detected. Although, vaccination against anthrax is characterized by an elevated protective profile and very low residual virulence, immunization with Sterne strains can cause abortion in cattle, presumably by the plasmid pX01 toxins in rare or special situations.(AU)

Este trabalho relata um aborto de um bovino, macho, Aberdeen Angus, por uma cepa vacinal de Bacillus anthracis, descreve os achados patológicos, microbiológicos e o sequenciamento do genoma. Os achados de necropsia incluíram áreas multifocais de hemorragias em diferentes órgãos. Histologicamente, órgãos afetados apresentaram hemorragia, exsudação de fibrina, necrose associada a miríades bacterianas bacilares e intenso infiltrado inflamatório neutrofílico. No exame microbiológico, foram isoladas numerosas colônias rugosas, não hemolíticas, cinzas e secas, com bordas irregulares a partir de amostras de fígado, pulmão e conteúdo do abomaso. A coloração de Gram revelou bastonetes Gram-positivos dispostos em cadeias. A identificação do B. anthracis foi confirmada pela detecção do marcador cromossômico molecular Ba813. Os genomas do isolado B. anthracis (SPV842_15) e do isolado vacinal (cepa vacinal brasileira), recuperado de uma vacina comercial utilizada na vaca prenhe, foram sequenciados. Comparações genômicas mostraram um elevado nível de identidade de nucleotídeos entre B. anthracis SPV842_15 e cepa vacinal brasileira (98,2%). Além disso, em ambas as estirpes foi detectada apenas a sequência do plasmídeo pX01. Embora a vacinação contra o antraz seja caracterizada por um perfil protetor elevado e uma virulência residual muito baixa, a imunização com estirpes de Sterne pode causar aborto em bovinos, presumivelmente pelas toxinas do plasmídeo pX01 em situações raras ou específicas.(AU)

Protective efficacy of commercial vaccines against a virulent field strain of Clostridium chauvoei / Eficácia protetiva de vacinas comerciais contra uma cepa de campo virulenta de Clostridium chauvoei

Blackleg is an acute and frequently fatal infection that mainly affects cattle and is caused by Clostridium chauvoei. Formalin-killed, whole-cell vaccines are commonly used to control blackleg. The aim of this study was to verify the protective efficacy of two commercial vaccines against the infection of guinea pigs with two strains of C. chauvoei, a virulent field strain (SBP 07/09) and the reference strain used in official tests (Manguinhos-Teixeira or MT). The strains used in the challenge were characterized by whole genome sequencing, and the minimal inhibitory concentrations of 15 antimicrobials were determined. To assess the protective efficacy, guinea pigs were vaccinated and subsequently challenged with C. chauvoei. All four vaccinated and challenged groups seroconverted after vaccination, while the control group remained seronegative, as determined by indirect ELISA. The identical performance of the two C. chauvoei strains in terms of virulence after challenge and their inability to infect vaccinated animals was correlated with their high genetic homology. Both commercial vaccines showed good protective efficacy against both the reference and field strains. Although C. chauvoei vaccination failures have been reported, the results from our study and others reported high similarity among C. chauvoei strains from all over the world, which suggests that the vaccine failures are not due to antigenic variability but inadequate vaccine management.(AU)

O carbúnculo sintomático é uma infecção aguda e frequentemente fatal que afeta principalmente os bovinos, causada pelo Clostridium chauvoei. A administração de vacinas formolizadas compostas pelas células bacterianas inteiras é comumente utilizada no controle desta doença. O objetivo deste estudo foi verificar, em cobaios, a eficácia protetiva de duas vacinas comerciais contra duas cepas de Clostridium chauvoei, uma cepa de campo virulenta (SBP 07/09) e a cepa de referência utilizada nos testes oficiais (MT, Manguinhos-Teixeira). Adicionalmente, realizar a caracterização das cepas utilizadas no desafio pelo sequenciamento completo do genoma e pela determinação da concentração inibitória mínima frente a 15 antimicrobianos. Para tanto, os cobaios foram vacinados e subsequentemente desafiados com. Os quatro grupos vacinados e desafiados soroconverteram, enquanto que o grupo controle permaneceu soronegativo pelo teste de ELISA indireto. O desempenho idêntico das duas cepas de C. chauvoei após o desafio in vivo e a incapacidade de infectar os animais vacinados está correlacionado com a homologia genética das cepas. Além disso, ambas as vacinas comerciais demonstraram uma adequada eficácia protetiva contra as cepas de campo e de referência. Embora o insucesso das vacinações contra C. chauvoei tenha sido previamente relatado, os resultados deste estudo, juntamente com a alta similaridade genética reportada previamente em cepas provenientes de diferentes continentes sugerem que as falhas vacinais não estão relacionadas à variabilidade antigênica, mas sim ao manejo vacinal inadequado.(AU)

Protective efficacy of commercial vaccines against a virulent field strain of Clostridium chauvoei / Eficácia protetiva de vacinas comerciais contra uma cepa de campo virulenta de Clostridium chauvoei

Blackleg is an acute and frequently fatal infection that mainly affects cattle and is caused by Clostridium chauvoei. Formalin-killed, whole-cell vaccines are commonly used to control blackleg. The aim of this study was to verify the protective efficacy of two commercial vaccines against the infection of guinea pigs with two strains of C. chauvoei, a virulent field strain (SBP 07/09) and the reference strain used in official tests (Manguinhos-Teixeira or MT). The strains used in the challenge were characterized by whole genome sequencing, and the minimal inhibitory concentrations of 15 antimicrobials were determined. To assess the protective efficacy, guinea pigs were vaccinated and subsequently challenged with C. chauvoei. All four vaccinated and challenged groups seroconverted after vaccination, while the control group remained seronegative, as determined by indirect ELISA. The identical performance of the two C. chauvoei strains in terms of virulence after challenge and their inability to infect vaccinated animals was correlated with their high genetic homology. Both commercial vaccines showed good protective efficacy against both the reference and field strains. Although C. chauvoei vaccination failures have been reported, the results from our study and others reported high similarity among C. chauvoei strains from all over the world, which suggests that the vaccine failures are not due to antigenic variability but inadequate vaccine management.

O carbúnculo sintomático é uma infecção aguda e frequentemente fatal que afeta principalmente os bovinos, causada pelo Clostridium chauvoei. A administração de vacinas formolizadas compostas pelas células bacterianas inteiras é comumente utilizada no controle desta doença. O objetivo deste estudo foi verificar, em cobaios, a eficácia protetiva de duas vacinas comerciais contra duas cepas de Clostridium chauvoei, uma cepa de campo virulenta (SBP 07/09) e a cepa de referência utilizada nos testes oficiais (MT, Manguinhos-Teixeira). Adicionalmente, realizar a caracterização das cepas utilizadas no desafio pelo sequenciamento completo do genoma e pela determinação da concentração inibitória mínima frente a 15 antimicrobianos. Para tanto, os cobaios foram vacinados e subsequentemente desafiados com. Os quatro grupos vacinados e desafiados soroconverteram, enquanto que o grupo controle permaneceu soronegativo pelo teste de ELISA indireto. O desempenho idêntico das duas cepas de C. chauvoei após o desafio in vivo e a incapacidade de infectar os animais vacinados está correlacionado com a homologia genética das cepas. Além disso, ambas as vacinas comerciais demonstraram uma adequada eficácia protetiva contra as cepas de campo e de referência. Embora o insucesso das vacinações contra C. chauvoei tenha sido previamente relatado, os resultados deste estudo, juntamente com a alta similaridade genética reportada previamente em cepas provenientes de diferentes continentes sugerem que as falhas vacinais não estão relacionadas à variabilidade antigênica, mas sim ao manejo vacinal inadequado.

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)

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)

Molecular characterization of virulence genes cctA, nanA, and fliC in Clostridium chauvoei from Rio Grande do Sul and São Paulo State, Brazil / Caracterização molecular dos genes de virulência cctA, nanA e fliC em Clostridium chauvoei isolados no Rio Grande do Sul e São Paulo, Brasil

Clostridium chauvoei toxin A (CctA), neuraminidase (NanA), and flagellin (FliC) proteins contribute to the pathogenicity of Clostridium chauvoei, the causative agent of blackleg in cattle. The aim of this study was to analyze the genetic variability of cctA, nanA, and fliC genes in C. chauvoei isolates from the Rio Grande do Sul and São Paulo state- Brazil, during different sampling periods. The presence of these genes was verified through PCR amplification and partial gene sequencing of 17 strains. Alignment of PCR amplicons combined with bioinformatics analysis was used in an attempt to study the variability across C. chauvoei solates. The similarity among the partial sequences of cctA and nanA genes was 100%. The sequencing of fliC revealed three different paralog alleles of flagellin, and two strains were seen to be polymorphic, with amino acid alterations in the predicted protein. Overall, this study indicates that strains of C. chauvoei isolated in Brazil are highly conserved with respect to the virulence factors evaluated.(AU)

Toxina A de Clostridium chauvoei (CctA), neuraminidase (NanA) e flagelina (FliC) são proteínas que contribuem para a patogenicidade de Clostridium chauvoei, o agente causador do carbúnculo sintomático em bovinos. O objetivo deste estudo foi analisar a variabilidade genética dos genes cctA, nanA, e fliC em C. chauvoei isolados em diferentes períodos no Rio Grande do Sul e São Paulo. A presença destes genes foi verificada pela amplificação dos produtos da PCR e sequenciamento parcial dos genes de 17 cepas. Os alinhamentos da amplificação dos produtos da PCR combinados com a análise de bioinformática foram utilizados na tentativa de avaliar a variabilidade dos genes entre os isolados de C. chauvoei. A similaridade do sequenciamento parcial dos genes cctA e nanA foi 100%. O sequenciamento do fliC revelou três alelos paralogos diferentes de flagelina e duas cepas mostraram polimorfismos, causando alterações na sequência de aminoácidos. As cepas de C. chauvoei isoladas no Brasil mostraram-se altamente conservadas em relação aos fatores de virulência avaliados neste estudo.(AU)

Early Detection of Bacillus anthracis From Saliva in Anticipation of a Bioterrorism Attack

Objective: To assess potential for early detection of oral infection by B. anthracis spores for preparedness of a bioterrorism attack. Material and Methods: The laboratory study used saliva with a range of initial anthrax concentrations, to compare detection by direct observation from conventional blood agar culture and by anthrax-specific PCR after a shorter culture in BHI broth. Three types of saliva were collected: stimulated saliva, unstimulated/whole saliva, and unstimulated/whole saliva with antibiotic treatment (for negative control). Using bivariate Kruskal-Wallis and Mann-Whitney tests for statistical analysis for factors that could affecting anthrax detection, significant differences between the test groups was assumed at p<0.05. Results: From unstimulated whole saliva heat shock treated at 62.50C, B. anthracis growth was detected with both methods. PCR detection from a BHI broth culture could shorten the time to diagnosis in comparison to conventional culture in blood agar. Conclusion: Saliva can provide useful samples for diagnosis of oropharyngeal anthrax. In comparison to conventional culture on blood agar, shorter-term culture in BHI broth provides potential for earlier detection and diagnosis.