Miocarditis por gram negativos / Gram-Negative Myocarditis

Resumen Objetivo: describir las características epidemiológicas, clínicas y microbiológicas de los pacientes que cursan con miocarditis por Enterobacterias. Métodos: se realizó una revisión sistemática de la literatura, en la que se incluyeron Pubmed, Ovid, Scopus, SciELO y LILACS sin exclusión por tipo de idioma. La población objetivo de estudio fueron los pacientes con diagnóstico de infección bacteriana por bacilo gram negativo mediante cultivo, técnicas moleculares o histopatología, y quienes presentaban biopsia de miocardio o, en su defecto, resonancia magnética cardiaca con hallazgos sugestivos de miocarditis. Resultados: se encontraron 742 artículos, de los cuales se incluyeron 24; en estos se reportaron 27 pacientes. La edad promedio fue de 31 años. El 81% de los pacientes eran de sexo masculino. El síntoma principal fue diarrea (80%), seguido de fiebre (53%) y dolor torácico (38%). El 37% de los pacientes fallecieron. El hallazgo más común en el electrocardiograma fue la elevación del segmento ST (36,7%). En quienes se realizó ecocardiograma se encontraron anormalidades en 50% de los casos, siendo más frecuente la disminución en la fracción de eyección. El microorganismo más común fue el Campylobacter jejuni, seguido por Salmonela sp. Conclusiones: la miocarditis causada por enterobacterias es más frecuente en pacientes adultos jóvenes de sexo masculino. Los síntomas gastrointestinales suelen estar presentes al momento de la presentación clínica. El diagnóstico requiere de alta sospecha clínica teniendo en cuenta que las anormalidades eléctricas y en ecocardiograma no se encuentran en todos los pacientes.

Abstract Objective: To describe the epidemiological, clinical, and microbiological characteristics of patients with myocarditis due to Enterobacteria. Methods: A systematic review was carried out on the literature, which included Pubmed, Ovid, Scopus, SciELO, and LILACS, with no exclusions due to language. The target population of the study were patients with a diagnosis of bacterial infection due to gram negative bacillus by means of a culture, or using molecular or histopathology technique. They also had to have had a myocardial biopsy or, if not, a cardiac magnetic resonance scan with findings suggestive of myocarditis. Results: Out of a total of 742 articles found, 24 of these, in which 27 patients were described, were included. The mean age was 31 years, and 81% were male. The main symptom was diarrhoea (80%), followed by fever (53%), and chest pain (38%). More than one-third (37%) of the patients died. The most common finding on the electrocardiogram (ECG) was elevation of the ST segment (36.7%). Abnormalities were found in 50% of the cases, on whom a cardiac ultrasound was performed, with a decrease in the ejection fraction being the most common. The most common microorganism was Campylobacter jejuni, followed by Salmonella spp. Conclusions: Myocarditis caused by enterobacteria is most common in young male patients. The gastrointestinal symptoms are usually present from the clinical onset. The diagnosis requires a high clinical suspicion, taking into account that the abnormalities in the ECG and cardiac ultrasound are not found in all patients.

COVID-19 and coinfection with Clostridioides (Clostridium) difficile in an infant with gastrointestinal manifestation / COVID-19 e coinfecção por Clostridioides (Clostridium) difficile em lactente com manifestações gastrintestinais

ABSTRACT We report the clinical case of an infant with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection with gastrointestinal signs and symptoms, predominantly vomiting. The patient also had colic, poor feeding, mild diarrhea and mild rhinorrhea without fever. The child had evidence of altered coagulation, increased interleukin 10, moderate dehydration and she was admitted to the pediatric intensive care unit. Simultaneously, the patient was diagnosed as Clostridioides difficile infection, which possibly may have facilitated the persistence of SARS-CoV-2 in feces, for more than 27 days, even after the nasopharyngeal test turned negative. This coinfection might exacerbate the gastrointestinal signs and symptoms and increased the possibility of fecal-oral transmission of SARS-CoV-2 and Clostridioides . The patient was breastfed and received complementary infant formula, hydrated with intravenous fluid, and was discharged without complications, 4 days after admission.

RESUMO Relatamos o caso clínico de uma lactente com infecção por coronavírus da síndrome respiratória aguda grave 2 (SARS-CoV-2) com sinais e sintomas gastrintestinais - predominantemente vômitos. A paciente apresentou, adicionalmente, cólica, dificuldade para mamar, evacuações amolecidas e rinorreia leve, sem febre. Houve evidências de alterações da coagulação, aumento de interleucina 10 e desidratação moderada, que justificaram internação na unidade de terapia intensiva. Simultaneamente, a paciente foi diagnosticada com infecção por Clostridioides difficile , que pode ter facilitado a persistência do SARS-CoV-2 nas fezes por mais de 27 dias, mesmo após negativação do teste nasofaríngeo. Essa coinfecção pode ter exacerbado os sinais e sintomas gastrintestinais e aumentado a possibilidade da transmissão do SARS-CoV-2 e Clostridioides . A paciente foi mantida em aleitamento materno e complemento com fórmula infantil, recebeu hidratação intravenosa e teve alta hospitalar, sem complicações, após 4 dias de internação.

Gut microbiota in Mexican patients with common variable immunodeficiency

Introduction: Common variable immunodeficiency (CVID) is the main symptomatic primary immunodeficiency and is associated with complex immune disorders. Gut microbiota interacts closely with the immune system, and intestinal dysbiosis is related to multiple diseases. Objective: To describe for the first time the composition of gut microbiota in Mexican patients with CVID. Methods: Fecal samples from five patients with CVID were collected and massive sequencing of the V3-V4 region of 16S rRNA gene was carried out using illumina technology. Results: Bacterial relative abundance was observed at all taxonomic levels. Firmicutes, Actinobacteria and Verrucomicrobia were the predominant phyla. The Clostridia class and the Clostridial order were the most common in their respective taxon; the Ruminococcaceae family predominated. A total of 166 genera were reported, with the most abundant being Faecalibacterium. Five species were identified, but only Bifidobacterium longum was present in all patients. Conclusions: Unlike healthy subjects' gut microbiota, where Firmicutes and Bacteroidetes predominate, the microbiota of the patients with CVID considered in this study was abundant in Firmicutes, Actinobacteria and Verrucomicrobia. The low presence of Bacteroidetes and high abundance of Firmicutes might indicate the existence of intestinal dysbiosis in these patients.

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)

Óleos essenciais e nanoemulsões como alternativas no controle de Clostridium sporogenes em mortadela / Essential oils and nanoemulsions as alternatives in the control of Clostridium sporogenes in mortadela

Avaliou-se a ação antimicrobiana da combinação dos óleos essenciais de canela, pimenta chinesa, limão Taiti e cardamomo e de suas nanoemulsões no controle de Clostridium sporogenes e sua influência sobre a oxidação lipídica em mortadela. A Concentração Mínima Bactericida (CMB) de cada óleo foi determinada pelo método de macrodiluição. As CMB encontradas foram: 0,10% óleo essencial de pimenta chinesa e canela e 1,00% óleo essencial de cardamomo e limão Taiti. A nanoemulsão elaborada apresentou redução significativa no número de células vegetativas de C. sporogenes na mortadela, além de ser o tratamento mais eficiente no controle da oxidação lipídica. Os resultados obtidos comprovam a ação antimicrobiana dos óleos testados, sugerindo sua possível utilização como conservantes naturais em produtos alimentícios.

Sinergismo de óleos essenciais sobre endósporos de Clostridium sporogenes / Essential oils sinergisms on Clostridium sporogenes endospores

A forma mais comum de inibir a produção de toxina botulínica em produtos cárneos cozidos é pela adição de sais de nitrito, o que pode gerar substâncias carcinogênicas (nitrosaminas), sendo desejável sua substituição. Os óleos essenciais vêm se destacando como agentes antimicrobianos, sendo interessante seu uso como conservante. O objetivo desse trabalho foi avaliar a ação sinergística de óleos essenciais sobre endósporos de Clostridium sporogenes, utilizado como modelo de pesquisa para C. botulinum. As concentrações mínimas esporicidas (CME) dos óleos de alecrim, tomilho, cravo, manjericão, ho wood e alho foram de 3% e de 0,375% para pimenta e canela. Os óleos de orégano e noz moscada não apresentaram ação esporicida nas concentrações testadas. Entre as combinações, as melhores foram de pimenta chinesa (0,1306%), alho (1,1%) e manjericão (1,1%) e pimenta chinesa (0,1306%), alho (1,1%) e tomilho branco (1,1%). Os resultados sugerem o sinergismo entre os óleos, sendo promissor seu uso em alimentos.

Epidemiología de bacterias anaerobias aisladas en muestras clínicas en el hospital san juan de dios, San José, Costa Rica, durante el trienio 2014, 2015 y 2016 / Epidemiology of anaerobic bacteria isolated in clinical samples at San Juan de Dios Hospital, San José, Costa Rica, during the 2014, 2015 and 2016 triennium

Resumen La epidemiología de las posibles poblaciones en riesgo de sufrir una infección por bacterias anaerobias a nivel nacional es desconocida, se debería de incentivar el conocimiento en los servicios de salud sobre este tipo de infecciones. Las bacterias anaerobias están relacionadas en los medios hospitalarios como causa importante de morbilidad, razón por la cual es conveniente conocer la epidemiologia y prevalecía de especies involucradas. En el Laboratorio de Bacteriología del Hospital San Juan de Dios, durante tres años, se analizaron un total de 1545 pacientes sospechosos de microorganismos anaerobios en medios prerreducidos, mediante un tamizaje se redujeron a un total de 469 posibles muestras, las cuales, fueron enviadas al Laboratorio de Investigación en Bacteriología Anaerobia (LIBA) para su correspondiente identificación. A lo largo de las semanas epidemiológicas de los tres años se encontraron en promedio de 1.77 casos confirmados / semana, con razón de sexo positiva a favor de los masculinos. Se determinó que solo 245 de las muestras enviadas presentaban uno o varios microorganismos anaerobios estrictos representando un 15.85% del total, identificándose 39 especies diferentes, en 306 cepas aisladas. Las mayormente importante fue el género Bacteriodes, provenientes de cavidad abdominal seguido de abscesos y heridas de piel. El presente estudio tiene como objetivo presentar datos que respalden la importancia clínica de la búsqueda de microorganismos anaerobios y que ayuden a los analistas de bacteriología a guiar cuales son los principales microorganismos esperables en muestras clínicas, además de conocer la prevalencia en general.

Abstract The epidemiology of the possible populations at risk of suffering an infection by anaerobic bacteria a national level is unknown, it should be encouraged the knowledge in the health services about this type of infections. Anaerobic bacteria are related in hospital environments as an important cause of morbidity, which is why it is convenient to know the epidemiology and prevalence of species involved. In the Bacteriology Laboratory of the Hospital San Juan de Dios, for three years, a total of 1545 patients suspected of anaerobic microorganisms in prereduced media were analyzed, through a screening was reduced to a total of 469 possible samples, which were sent to the Anaerobic Bacteriology Research Laboratory (LIBA) for its corresponding identification. Throughout the epidemiological weeks of the three years were found on average of 1.77 confirmed cases / week, with a positive sex ratio in favor of men. It was determined that only 245 of the samples sent had one or several strict anaerobic microorganisms representing 15.85% of the total, identifying 39 different species, in 306 isolated strains. The most important was the genus Bacteriodes, coming from the abdominal cavity followed by abscesses and skin wounds. The present study aims to present data that support the clinical importance of the search for anaerobic microorganisms and that help the analysts of bacteriology to guide which are the main expected microorganisms in clinical samples, in addition to knowing the prevalence in general.

Intereactions between doripenem and clavulanate ­ Application of minimal inhibitory concentration analysis and cytometry flow for bactericidal studies

Background: In view of the current low efficacy of bacterial infection treatment the common trend towards searching for antibiotic systems exhibiting synergistic action is well justified. Among carbapenem analogues a particularly interesting option is provided by combinations of clavulanic acid with meropenem, which have proven to be especially effective. Results: Determination of the minimal inhibitory concentration (MIC) along with the method based on flow cytometry constitutes an important tool in the identification of bacterial sensitivity to active substances. Within this study the inhibitory effect of doripenem, clavulanic acid and the doripenem-clavulanate acid system was analyzed in relation to such bacteria as Salmonella enteritidis, Salmonella typhimurium, Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris, Clostridium butyricum and Clostridium pasteurianum, Acinetobacter baumannii, Enterobacter aerogenes. The lowest MIC, amounting to 0.03 µg/mL, was observed for the doripenem-clavulanate acid system in the case of E. coli ATCC 25922. In turn, the lowest MIC for doripenem applied alone was recorded for K. pneumoniae ATCC 31488, for which it was 0.1 µg/mL. The strain which proved to be most resistant both to doripenem and the doripenem-clavulanate acid system, was A. baumannii, with MIC of 32 µg/mL (clinical isolate) and 16 µg/mL (reference strain). Cytometric analysis for P. aeruginosa ATCC 27853 and S. aureus ATCC 25923 showed changes in cells following exposure to limiting concentrations of the active substance. Conclusions: Analysis of MIC supplies important information concerning microbial sensitivity to active substances, mainly in terms of limiting concentrations causing mortality or vitality of the tested species, which is essential when selecting appropriate antibiotic therapy.

Diarreia causada por clostridium difficile: recentes avanços / Diarrhea caused by clostridium difficile: recent Advances

A infecção causada por Clostridium difficile (C. difficile), um dos agentes causadores de diarréia aguda e recorrente, tem como principal fator de risco o uso de antimicrobianos. Recentemente, houve um aumento da incidência e da mortalidade desta afecção. Clinicamente, a mesma pode manifestar-se desde um quadro de diarreia aquosa leve até a forma grave de colite pseudomembranosa. O objetivo deste artigo é apontar as mudanças epidemiológicas da infecção pelo C. difficile, além de rever fatores de risco, manifestações clínicas, métodos diagnósticos, tratamento e prevenção desta infecção. O aumento na gravidade da infecção causada pelo C. difficile é relacionado a uma nova cepa hipervirulenta, BI/NAPI/Ribotipo 027, que apresenta maior capacidade de produção de toxinas. Essa nova cepa, mais virulenta, ainda não foi detectada no Brasil, porém como já foi identificada em outros países da América, alerta para a preocupante capacidade de disseminação universal. Essa revisão é baseada em artigos publicados nos últimos 10 anos, utilizando como base de dados o PubMed e o Scielo (Scientific Eletronic Library Online), com as palavras-chave: Epidemiologia, diarreia, Clostridium difficile e cepa hipervirulenta.

Clostridium difficile (C. difficile) is a causative agent of diarrhea and its main risk factor is the use of antimicrobials. Recently, there was an increase in incidence and mortality. Clinical symptoms can manifest from mild watery diarrhea to severe pseudomembranous colitis. The purpose of this article is to make a literature review of C. difficile -associated diarrhea including its recent epidemiological changes. The increase in the severity of infection caused by Clostridium difficile was related to a new hypervirulent strain, BI/NAPI/ribotype 027, with greater capacity for production of toxins, responsible for clinical manifestations. This new strain, more virulent, has not yet been detected in Brazil, but it was already identified in other countries of America, warns the disturbing ability to universal dissemination. This review is based on articles published in the last 10 years, using as database PubMed and Scielo (Scientific Electronic Library Online), with the keywords: Epidemiology, diarrhea, Clostridium difficile and hypervirulent strain.

Improvement of hydrogen yield of ethanol-producing Escherichia coli recombinants in acidic conditions

Background: An effective single culture with high glycerol consumption and hydrogen and ethanol coproduction yield is still in demand. A locally isolated glycerol-consuming Escherichia coli SS1 was found to produce lower hydrogen levels under optimized ethanol production conditions. Molecular approach was proposed to improve the hydrogen yield of E. coli SS1 while maintaining the ethanol yield, particularly in acidic conditions. Therefore, the effect of an additional copy of the native hydrogenase gene hycE and recombinant clostridial hydrogenase gene hydA on hydrogen production by E. coli SS1 at low pH was investigated. Results: Recombinant E. coli with an additional copy of hycE or clostridial hydA was used for fermentation using 10 g/L (108.7 mmol/L) of glycerol with an initial pH of 5.8. The recombinant E. coli with hycE and recombinant E. coli with hydA showed 41% and 20% higher hydrogen yield than wild-type SS1 (0.46 ± 0.01 mol/mol glycerol), respectively. The ethanol yield of recombinant E. coli with hycE (0.50 ± 0.02 mol/mol glycerol) was approximately 30% lower than that of wild-type SS1, whereas the ethanol yield of recombinant E. coli with hydA (0.68 ± 0.09 mol/mol glycerol) was comparable to that of wild-type SS1. Conclusions: Insertion of either hycE or hydA can improve the hydrogen yield with an initial pH of 5.8. The recombinant E. coli with hydA could retain ethanol yield despite high hydrogen production, suggesting that clostridial hydA has an advantage over the hycE gene in hydrogen and ethanol coproduction under acidic conditions. This study could serve as a useful guidance for the future development of an effective strain coproducing hydrogen and ethanol.

Tétano canino ­ Relato de caso / Tetanus in dogs ­ Case report / Tétano canino ­ Relato de caso

O tétano refere-se a uma condição infecciosa ocasionada por toxinas do Clostridium tetani, bacilo Gram positivo e anaeróbico, o qual ao sofrer lise e morte nos tecidos do hospedeiro libera toxinas capazes de bloquear a liberação do neurotransmissor inibidor de glicina e a liberação do ácido gama-amino butírico, levando a um quadro de tetania. Os sinais clínicos da enfermidade são o trismo e opistótono, entre outros. O diagnóstico se dá por meio de histórico e sinais clínicos e o tratamento é sintomático. O presente artigo tem o objetivo de relatar um caso clínico de tétano em um macho canino da raça Pitbull, de um ano de idade. O cão foi, atendido no Hospital Veterinário da Universidade Paranaense, Umuarama, PR, com sinais clínicos neurológicos como rigidez muscular e opistótono, iniciados após ferimento no coxim plantar. Pelo histórico e sinais clínicos firmou-se o diagnóstico de tétano. Foi instituído tratamento, mas o animal teve óbito, sendo que a doença tem alta mortalidade na espécie.(AU)

Tetanus is an infectious condition caused by Clostridium tetani toxins, a Gram-positive, anaerobic bacillus, which, when undergoing lysis and death in the host tissues, releases toxins capable of blocking the release of the glycine inhibitory neurotransmitter and the release of gamma-amino butyric acid, leading to a tetanic state. The clinical signs of the disease are trismus and opisthotonus, among others. The diagnosis is made by means of history and clinical signs, with symptomatic treatment. This article aims to report a clinical case of tetanus in a one-year-old Pitbull male canine. The dog was attended at the Veterinary Hospital at Universidade Paranaense, in the city of Umuarama, Paraná, presenting neurological clinical signs such as muscular and opisthotonous rigidity, initiated after injury to the plantar cushion. The diagnosis of tetanus was confirmed by the history and clinical signs. Treatment was instituted but the animal died, since the disease present a high mortality rate in the species.(AU)

El tétano se refiere a una condición infecciosa ocasionada por toxinas de Clostridium tetani, bacilo Gram positivo y anaeróbico, el cual al sufrir lise y muerte en los tejidos del hospedero suelta toxinas capaces de bloquear la liberación del neurotransmisor inhibidor de glicina, y la liberación del ácido gama amino butírico, llevando a un cuadro de tetania. Los signos clínicos de la enfermedad son el trismo y opistódomo, entre otros. El diagnóstico se da por medio de histórico y signos clínicos y el tratamiento es sintomático. El presente artículo tiene el objetivo de relatar un caso clínico de tétano en un canino macho de la raza Pitbull, de un año de edad. El perro ha sido atendido en el Hospital Veterinario de la Universidad Paranaense, Umuarama PR, con señales clínicos neurológicos como rigidez muscular y opistódomo, iniciados tras herimiento en el coxin plantar. Por el histórico y señales clínicos se ha llegado al diagnóstico de tétano. Se empezó el tratamiento, pero el animal vino a óbito, siendo que la enfermedad tiene alta mortalidad en la especie.(AU)

Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry for identification of Clostridium species isolated from Saudi Arabia

Abstract The aim of this study was to identify different Clostridium spp. isolated from currency notes from the Ha’il region of Saudi Arabia in September 2014 using MALDI–TOF-MS. Clostridium spp. were identified by Bruker MALDI–TOF-MS and compared with VITEK 2. The confirmation of the presence of different Clostridium spp. was performed by determining the sequence of the 16S ribosomal RNA gene. In this study, 144 Clostridium spp. were isolated. Among these specimens, MALDI–TOF-MS could identify 88.8% (128/144) of the isolates to the species level and 92.3% (133/144) to the genus level, whereas, VITEK 2 identified 77.7% of the (112/144) isolates. The correct identification of the 144 isolates was performed by sequence analysis of the 500 bp 16S rRNA gene. The most common Clostridium spp. identified were Clostridium perfringens (67.36%), Clostridium subterminale (14.58%), Clostridium sordellii (9%) and Clostridium sporogenes (9%). The results of this study demonstrate that MALDI–TOF-MS is a rapid, accurate and user friendly technique for the identification of Clostridium spp. Additionally, MALDI–TOF-MS has advantages over VITEK 2 in the identification of fastidious micro-organisms, such as Clostridium spp. Incorporating this technique into routine microbiology would lead to more successful and rapid identification of pathogenic and difficult to identify micro-organisms.

ELISA and modified toxin-binding inhibition test for quality control of the clostridial vaccine processes / ELISA direto e ToBI-test modificado para o controle de qualidade de processos de vacinas clostridiais

This study aimed to assess and standardize the ELISA and modified ToBI test in vitro methods in order to verify the potency of epsilon toxicoid in comparison with the in vivo TCP method. The following epsilon toxoids were used: NIBSC standard from batches 375/07, 532/08, 551/08, 373/07 and 378/07. These were evaluated using a TCP test, ELISA and ToBI tests. The results indicate that the correlation ratio between the dilutions of standard NIBSC toxicoid and absorbance values of 89.44% obtained with the ELISA method support the use of the curve to evaluate epsilon toxoids. However, it was observed that the absorbance values were similar for all toxoids, thus presenting no significant difference between higher and lower concentration toxoids. For the ToBI test, the correlation ratio of 96.76, obtained in the curve pattern, demonstrates the effectiveness of the curve to be used in the epsilon toxoid evaluation. The correlation ratio between the titration degrees of toxoids obtained through TCP and ToBI tests was higher than 90%. It is concluded that the type of ELISA test used does present discriminative power for toxoids with different concentrations, which does not support the use of this technique for such a purpose. The ToBI test can be used as a screening method for it is sensitive and effective to detect epsilon toxicoid produced by C. perfringens type D...

Teve-se por objetivo avaliar e padronizar as metodologias in vitro, ELISA e ToBI-test modificado, para a análise de toxoide épsilon, em comparação com a metodologia in vivo TCP. Foram utilizados os seguintes toxoides épsilon: padrão NIBSC e os lotes 375/07, 532/08, 551/08, 373/07 e 378/07, os quais foram avaliados por métodos in vivo, TCP, e in vitro, ELISA e ToBI-test. A análise do título de toxoide épsilon por meio dos métodos in vitro foi realizada a partir de uma curva-padrão, estabelecida previamente. Os principais resultados mostram que os valores de absorbância foram semelhantes para todos os toxoides, não apresentando diferença significativa entre os toxoides mais concentrados e menos concentrados. No ToBI-test, o coeficiente de correlação de 96,76%, obtido na curva-padrão, demonstra a eficiência da curva para avaliação do toxoide épsilon. O coeficiente de correlação entre os títulos de toxoide obtidos pelo TCP e ToBI-test foi superior a 90%. Conclui-se que o tipo de ELISA utilizado não apresenta poder discriminativo para toxoides com diferentes concentrações, inviabilizando a técnica para esse fim. O ToBI-test pode ser utilizado como um método de triagem sensível e eficaz para a detecção de toxoide épsilon de C. perfringens tipo D...

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.

Kinetics of epsilon antitoxin antibodies in different strategies for active immunization of lambs against enterotoxaemia / Cinética dos anticorpos antitoxina epsilon em diferentes estratégias de imunização ativa em cordeiros contra a enterotoxemia

Enterotoxaemia, a common disease that affects domestic small ruminants, is mainly caused by the epsilon toxin of Clostridium perfringens type D. The present study tested four distinct immunization protocols to evaluate humoral response in lambs, a progeny of non-vaccinated sheep during gestation. Twenty-four lambs were randomly allocated into four groups according to age (7, 15, 30 and 45 days), receiving the first dose of epsilon toxoid commercial vaccine against clostridiosis with booster after 30 days post vaccination. Indirect ELISA was performed after the first vaccine dose and booster to evaluate the immune response of the lambs. Results showed that for the four protocols tested all lambs presented serum title considered protective (≥0.2UI/ml epsilon antitoxin antibodies) and also showed that the anticipation of primovaccination of lambs against enterotoxaemia conferred serum title considered protective allowing the optimization of mass vaccination of lambs.

Enterotoxemia, uma das mais comuns enfermidades que acomete os pequenos ruminantes domésticos, é causada principalmente pela toxina épsilon de Clostridium perfringens tipo D. O presente estudo avaliou a resposta humoral conferida por quatro protocolos distintos de primovacinação na progênie de ovelhas não vacinadas durante a gestação. Vinte e quatro cordeiros foram aleatoriamente divididos em quatro grupos de acordo com a idade (dias) que receberam a primeira dose da vacina comercial contra clostridiose contendo toxóide epsilon na sua formulação. Todos os cordeiros foram vacinados aos 7, 15, 30 ou 45 dias de idade e receberam um reforço da dose 30 dias após a vacinação. A avaliação sorológica dos cordeiros pelo teste de ELISA indireto foi realizada por ocasião da administração da primeira dose da vacina. Os resultados elucidaram não haver comprometimento da resposta imune de cordeiros vacinados tanto aos 7, 15, 30 ou 45 dias de idade associada ao reforço da dose 30 dias após, demonstrando assim que a antecipação da primeira vacinação conferiu proteção aos cordeiros contra a enterotoxemia, permitindo otimizar o planejamento da vacinação em massa dos cordeiros.

Evaluating Colombian Clostridium spp. Strains hydrogen production using glycerol as substrate

Background: The ability for hydrogen production of 13 native strains of Clostridium spp. strains isolated from Colombian soil was evaluated using glycerol substrate. Glycerol to hydrogen conversion was investigated using a batch fermentation reactor and industrial glycerol source (50 g.l-1, pH 7.00). Results: The results were quantified regarding acids, hydrogen, biomass and solvent production. The selected strain gave good hydrogen over production output at 14.4 mmol H2.l-1, productivity 0.3167 mg H2.h-1 l-1 culture mediumand yield 0.1962 mol H2.mol-1 glycerol. A further fermentation assay a 4.0 liter batch reactor let to being 0.26 mg.l-1.h-1 after 18 hrs of fermentation. Logistic model, Luedeking-Piret model and Luedeking-Piret modified models were used for modeling changes in cell growth, hydrogen production and substrate consumption (Correlation coefficients R² = 0.95 for biomass substrate, R² = 0.77 hydrogen production). Conclusions: Our results indicate that hydrogen production through glycerol bioconversion by native strains is possible.

Formación de endosporas en Clostridium y su interacción con el proceso de solventogénesis / Endospore formation in Clostridium and its interaction with solventogenesis

La solventogénesis y la esporulación son mecanismos de las células de Clostridium para resistir ambientes hostiles. Este segundo proceso ha sido estudiado utilizando como modelo lo que sucede con Bacillus, aunque se reconocen diferencias marcadas entre los dos géneros especialmente en el inicio, específicamente en los eventos por medio de los que se da la fosforilación del regulador maestro Spo0A. En la actualidad se ha avalado la teoría que afirma que tres histidin quinasas huérfanas, diferentes a las proteínas Spo0B (histidin quinasa) y Spo0F(fosfotransferasa) en Bacillus, son las encargadas de fosforilar en Clostridium de forma directa a Spo0A, que posteriormente activa la transcripción de diferentes factores sigma relacionados, de forma similar en los dos géneros bacterianos. La proteína Spo0A, perteneciente a la familia de reguladores de respuesta, se comporta como una entidad regulatoria global que tiene injerencia sobre los procesos de formación de esporas y solventes, modulando genes necesarios para que se produzcan acetona y butanol, además de genes de esporulación. El entendimiento de este proceso ha llevado a los investigadores a emplear diferentes técnicas moleculares que permitan incrementar la producción de solventes, así como eliminar la propiedad de las células de producir endosporas. Por tal razón, este escrito presenta un resumen de estas dos redes de expresión de genes, conectadas por el regulador maestro Spo0A.

Solventogenesis and sporulation are mechanisms used by Clostridium cells to resist hostile environments. Sporulation has been studied using as a model what happens with Bacillus, but marked differences were recognized, particularly in the events that led the phosphorylation of the master controller Spo0A. Currently, a theory that claims that three orphan histidine kinases, different from Spo0B (histidin kinase) and spo0F (phosphotransferase) proteins in Bacillus, phosphorilate directly Spo0A in Clostridium activating transcription of different sigma factors which are similar in the two bacterial genera, has been supported. Spo0A protein, which belongs to the family of response regulators, behaves as an entity that has global regulatory interference on the processes of spore formation and solvents, modulating genes necessary to produce acetone and butanol. The understanding of this process has led researchers to employ different molecular techniques that increase the production of solvents, and remove the property of the cells to produce endospores. Reason why, this paper presents an updated summary of these two gene expression networks, connected by the master regulator spo0A.

Óleo essencial de orégano, alecrim, canela e extrato de pimenta no controle de Salmonella, Eimeria e Clostridium em frangos de corte / Oregano, rosemery, cinnamon essential oil and pepper extract to control Salmonella, Eimeria and Clostridium in broiler chickens

Este trabalho foi conduzido com o objetivo de avaliar a eficiência de um composto vegetal contendo óleo essencial de orégano, alecrim, canela e extrato de pimenta vermelha no controle de Salmonella, Eimeria e Clostridium em frangos de corte. Para tal, foram realizados dois experimentos. No primeiro avaliou-se a eficiência deste produto no controle de Clostridium perfringens após desafio com Eimeria acervulina, E. maxima e E. tenella. Aves de um dia de idade foram divididas em três grupos: T1 - dieta controle sem aditivo promotor de crescimento; T2 - dieta com adição de avilamicina (10ppm); e T3 - dieta com adição do composto vegetal (100ppm). O uso do composto vegetal na alimentação de frangos reduziu lesões específicas de E. maxima e E. tenella aos 14 dias pós-inoculação (PI) como também reduziram a contagem de unidades formadoras de colônias (UFC) de Clostridium perfringens no conteúdo do ceco das aves em relação ao grupo controle. No segundo experimento avaliou-se a eficiência deste mesmo produto em aves desafiadas com Salmonella Enteritidis. Aves de um dia de idade foram distribuídas em três tratamentos, sendo T1 - dieta controle sem adição de antibiótico promotor de crescimento, T2 - dieta com 10ppm de Avilamicina, T3 - dieta com 100ppm de um produto a base do composto vegetal acima citado. Aos 21 dias de idade todas as aves foram inoculadas com 10(5) UFC de Salmonella Enteritidis. A utilização do composto vegetal e avilamicina diminuiu a excreção de Salmonella nas aves 72 horas PI de Salmonella. A utilização do composto vegetal aumentou a relação vilo/células CD3+ no duodeno, em relação ao grupo avilamicina e controle, porém não teve efeito sobre a expressão destas células no ceco.

The efficiency of a product in broiler feed containing essential oil of oregano, rosemary, cinnamon and extract of red pepper (plant compost) in the control of Salmonella, Eimeria and Clostridium was evaluated. Two experiments were carried out to evaluate the product. In the first experiment the efficiency of this product to control Clostridium perfringens after challenge with Eimeria acervulina, E. maxima and E. tenella was assessed. Day old chicks were allotted into three groups: T1 - control diet without growth promoter, T2 - diet with avilamycin (10ppm), and T3 - diet with addition of the plant compost (100ppm). The use of the plant compost in broiler diets reduced specific lesions of E. maxima and E. tenella at 14 days after inoculation and reduced the count of colony forming units (CFU) of Clostridium perfringens in the ceca comparing to the control group. In the second trial the efficiency of the same product in birds challenged with Salmonella Enteritidis was evaluated. Day old birds were submitted to three experimental diets: T1 - control diet without antibiotics growth promoter, T2 - diet with 10ppm Avilamycin, T3 - diet with 100ppm of the plant compost mentioned above. At 21 days of age all birds were inoculated with 10(5) CFU of Salmonella Enteritidis. The use of the plant compost and avilamycin decreased the excretion of Salmonella in poultry 72 hours after the inoculation. The use of the plant compost increased villous/CD3+ cells in the duodenum, compared to group avilamycin and control, but had no effect on the expression of these cells in the cecum.