Surto de botulismo em bovinos leiteiros associado ao consumo de resíduo de fecularia: relato de caso / Botulism outbreak in dairy cattle associated with cassava by-product intake: case report / Brote de botulismo en ganado lechero asociado al consumo de residuos de almidón: informe de un caso

O resíduo de fecularia é um subproduto da agroindústria utilizado com frequência na alimentação de bovinos leiteiros pelo alto teor energético e baixo custo. No entanto, possui características propícias ao desenvolvimento de microrganismos patogênicos como o Clostridium botulinum, causador do botulismo. Este relato é o primeiro na literatura consultada a descrever um surto de botulismo em uma propriedade de bovinos leiteiros devido ao consumo de massa de mandioca, alimento ofertado em muitas propriedades do Oeste do Paraná. 16 animais, de um total de 24 morreram em um período de 14 dias, com sinais clínicos de respiração abdominal, sialorreia, paresia de membros pélvicos, decúbito esternal seguido de lateral permanente, diminuição dos tônus mandibular e lingual e morte em até 48 horas. Os animais não eram vacinados para clostridioses e recebiam o resíduo em cocho coletivo. O diagnóstico foi baseado no histórico, na avaliação das condições de armazenamento do alimento na propriedade, no exame físico e na ausência de lesões macroscópicas e histológicas. Conclui-se que o resíduo de fecularia deteriorado é um substrato propício para desenvolvimento do C. Botulinum sendo um alimento passível de provocar surtos de botulismo em bovinos leiteiros com alta morbidade e letalidade.

Cassava is an industry by-product waste used to feeding cattle due to its low cost and high energy. However, this food presents characteristics to development of pathogenic microorganisms such as Clostridium botulinum causer of botulism. This disease results from the ingestion of preformed neurotoxins that act at the neuromuscular junctions causing flaccid paralysis of striated muscles. This is the first report to describe botulism due the intake of cassava by-product (tapioca waste), a common food offered in the west of Parana State. 16 dairy cattle died from a total of 24 animals in 14 days. The clinical signs were dyspnea with abdominal pattern, sialorrhea, pelvic limb paresis, decreased mandibular and tongue tonus and permanent sternal progressing to lateral decubitus and death until 48 hours after the onset of signs. The diagnosis was based on anamnesis, evaluation of food storage conditions, clinical examination, and the absence of macroscopic and histology lesions. It was concluded that cassava by-product has conditions to the development of Clostridium botulinum and can cause botulism outbreak with high morbidity and lethality.

El residuo de almidón es un subproducto de la agroindustria que suele utilizarse para alimentar al ganado lechero por su alto contenido energético y su bajo coste. Sin embargo, tiene características favorables para el desarrollo de microorganismos patógenos como Clostridium botulinum, causante del botulismo. Este informe es el primero en la literatura que describe un brote de botulismo en una hacienda de ganado lechero debido al consumo de pasta de mandioca, alimento ofrecido en muchas propiedades del oeste de Paraná. 16 animales, de un total de 24, murieron en un período de 14 días, con signos clínicos de respiración abdominal, sialorrea, paresia de los miembros pélvicos, decúbito esternal seguido de decúbito lateral permanente, disminución del tono mandibular y lingual y muerte en 48 horas. Los animales no estaban vacunados contra la clostridioses y recibían sus desechos en un comedero comunitario. El diagnóstico se basó en la anamnesis, la evaluación de las condiciones de almacenamiento de los alimentos en la propiedad, el examen físico y la ausencia de lesiones macroscópicas e histológicas. Se concluye que el residuo de almidón descompuesto es un sustrato favorable para el desarrollo de C. botulinum y es un alimento que puede causar brotes de botulismo en ganado lechero con alta morbilidad y letalidad.

Type C botulism in dogs from rural properties located in Goiânia, Brazil

Botulism is a disease usually fatal, caused by the ingestion of neurotoxins produced by Clostridium botulinum. In dogs, intoxication is caused by the ingestion of botulinum toxin type C, and animals often recover spontaneously. The present study describes the occurrence of type C botulism in two dogs domiciled on neighboring rural properties in the municipality of Goiânia, state of Goiás, Brazil, probably associated with ingestion of decomposing bovine carcass. Upon clinical evaluation, the dogs were alert in the lateral decubitus position with ascending flaccid paralysis, absence of eyelid reflexes, and reduced muscle tone. Due to their worsening clinical symptoms, the animals died within 12 h and 3 days after supportive treatment. Botulinum toxin type C was identified, in the serum and feces of both dogs, by seroneutralization in mice with homologous monovalent antitoxin. The results of the high-throughput gene sequencing showed that the abundance of C. botulinum in the fecal microbiota of one of the affected dogs was low (0.53%). In this way, the present study highlights the need of sanitary practices related to the appropriate collection and disposal of bovine carcasses in rural areas since they represent a risk factor for the occurrence of botulism in dogs domiciled on rural properties.

A toxina botulínica como alternativa do arsenal terapêutico na odontologia / Botulinum toxin as alternative of therapeutical arsenal in dentistry

A evolução dos materiais, técnicas e fármacos culminou com a descoberta e uso da toxina botulínica Tipo A na Medicina e na Odontologia. Longe do escopo e da discussão sobre seu uso cosmético ou funcional, a toxina pode ser empregada em múltiplas terapias no terço inferior da face. A metodologia utilizada foi uma revisão sistemática sobre artigos científicos em revistas e periódicos indexados e de fácil aquisição nas bases de dados PubMed/MEDLINE, Lilacs, BBO, SciELO e Google Acadêmico nos últimos 10 anos, acrescidos de referências clássicas e estudos considerados relevantes para essa publicação. A toxina botulínica é considerada um procedimento minimamente invasivo clássico, pois não é agressiva e nem penetra o organismo de forma extensiva, com ato operatório que não exige uma hora clínica longa e permite o retorno do paciente às suas atividades laborais de maneira bastante precoce. Empregada na Odontologia: na sialorreia, nas assimetrias faciais com origem muscular, nas disfunções e nos problemas da articulação temporomandibular, no bruxismo, no controle do sorriso gengival, na distonia orofacial, no controle da força muscular após aposição de fixações em Implantodontia, entre outros. Foi possível concluir que a toxina botulínica é uma alternativa promissora dentro do arsenal terapêutico do cirurgião-dentista devido ao seu múltiplo emprego, com possibilidade de correções e reversibilidade superiores quando comparadaa técnicas cirúrgicas, por exemplo, com menor índice de morbidade, maior conforto e resultado imediato para o paciente

The evolution of materials, techniques and drugs culminated in the discovery and use of Botulinum Toxin, Type A, in Medicine and Dentistry. Far from the scope and discussion of its cosmetic or functional use, the toxin can be employed in multiple therapies in the lower third of the face. The methodology used was a systematic review of scientific articles in indexed and easily retrieved journals and papers in the PubMed/ MEDLINE, Lilacs, BBO, SciELO and Google Academic databases in the last 10 years, along with classic references and studies considered relevant to this publication. Botulinum toxin is considered a classic minimally invasive procedure, since it is neither aggressive nor penetrating the organism extensively, with an operative procedure that doesn´t require a long clinical time and allows the patient to return to his work activities in a very precocious way. Employed in Dentistry: in sialorrhoea, facial asymmetries with muscular origin, dysfunctions and temporomandibular joint problems, bruxism, gingival smile control, orofacial dystonia, muscle strength control after fixations placement in Implant Dentistry, among others. It was possible to conclude that botulinum toxin is a pr omising alternative within the therapeutic arsenal of the dentist due to its multiple use, with the possibility of superior corrections and reversibility when compared to surgical techniques, for example, with lower morbidity index, greater comfort and immediate result for the patient.

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.

Uso da toxina botulínica na correção do sorriso gengival / Use of botulinal toxin in the correction of gengival smile

Introdução: A beleza do sorriso representa uma expressão primordial na harmonização da face, e está diretamente ligada à autoestima do indivíduo. A atividade do sorriso é determinada pela contração de músculos específicos, e pode apresentar diferentes níveis de exposição gengival. Quando maior que 3mm é denominado sorriso gengival e gera uma condição esteticamente desagradável. Entre as suas diversas causas, a hiperatividade muscular se destaca. Objetivo: apresentar uma revisão de literatura a respeito da toxina botulínica, que se tornou uma ferramenta bastante utilizada como método complementar ou exclusivo na correção do sorriso gengival. Método: Para a pesquisa, foram utilizados os seguintes descritores: toxina botulínica tipo A, estética dentária e clostridium botulinum, utilizando os operadores booleanos AND e OR, inclusos artigos em língua portuguesa, inglesa e espanhola para descrever seu histórico, mecanismo de ação, durabilidade, indicações, contraindicações e técnica de aplicação. Conclusão: Conclui-se que o seu emprego como método de tratamento conservador pelo cirurgião-dentista é viável desde que possua conhecimento da anatomia facial, interações entre os músculos e farmacologia da neurotoxina, garantindo assim segurança, melhoria na estética do sorriso e satisfação do paciente.

Introduction: The beauty of the smile represents a primordial expression in the harmonization of the face and is directly linked to the self-esteem of the individual. Smile activity is determined by the contraction of specific muscles and may present different levels of gingival exposure. When bigger than 3mm, it is called gingival smile, and generates an aesthetically unpleasant condition. Among its many causes, muscular hyperactivity is highlighted. Objective: The objective of this work was to present a literature review about botulinum toxin, which has become a widely used tool as a complementary or exclusive method for gingival smile correction. Method: The following descriptors were used: Botulinum toxin type A, dental aesthetics and clostridium botulinum, using Boolean operators AND and OR, including articles in Portuguese, English and Spanish to describe their history, mechanism of action, durability, indications, contraindications and application technique. Conclusion: It is concluded that its use as a method of conservative treatment by the dental surgeon is feasible if it has a thorough knowledge of facial anatomy, interactions between muscles and neurotoxin pharmacology, thus ensuring safety, improvement in smile aesthetics and patient satisfaction.

Three-dimensional assessment of enamel and dentine in mouse molar teeth during masseter muscle hypofunction / Evaluación tridimensional del esmalte y la dentina de dientes molares de ratones durante la hipofunción del músculo masetero

Background: Mouse molar is a widely used model for teeth development. However, the effect of masticatory function on enamel and dentine in adult individuals remains poorly understood. As reported, the unilateral masseter hypofunction induced by botulinum toxin type A (BoNTA) resulted in mandibular bone damage and signs of unilateral chewing in adult mice. Objective: We aimed to assess the amount of enamel and dentine in the first molar (M1) during the unilateral masseter hypofunction in mice, using high-resolution X-ray microtomography (µCT) as threedimensional approach. Materials and methods: Mandibles of adult BALB/c mice, located either in a Control-group (without intervention) or a BoNTA-group, were ex-vivo scanned using µCT. Treated individuals received each one BoNTA intervention in the right masseter, and saline solution in the left masseter (intra-individual control). Enamel and dentine from M1 were segmented, and volume, thickness and mesial root length were quantified. Results: Enamel volume from treated side resulted unchanged after 2 weeks of unilateral masseter hypofunction. No differences for enamel volume were found between both sides of control individuals, and between these and samples from hypofunctional side in BoNTA-group. Enamel volume from saline-injected side was reduced when compared with experimental side (p<0,01). No differences in dentine volume, thickness of enamel and dentine, and mesial root length were found for any group. Conclusion: The amount of enamel in hypofunctional molars remains unaffected after unilateral BoNTA intervention in the masseter, but contralateral side showed reduced enamel volume. Therefore, increased functional wearing during unilateral chewing after BoNTA intervention should be considered.

Introducción: El molar de ratón es utilizado como modelo de estudio en el desarrollo dental. El efecto de la función masticatoriasobre el tejido dental en individuos adultos aún se comprende. En ratones adultos, la hipofunción unilateral del masetero inducida por toxina botulínica tipo A (BoNTA) resultó en daño óseo mandibular y signos de masticación unilateral. Objetivo: Evaluamos la cantidad de esmalte y dentina en el primer molar (M1) durante la hipofunción unilateral del músculo masetero en ratones mediante análisis con microtomografía (µCT). Materiales y métodos: Las mandíbulas de ratones BALB/c adultos, del grupo Control (sin intervención) o el grupo BoNTA, fueron escaneadas ex-vivo con µCT. Los individuos tratados se inyectaron con BoNTA en el masetero derecho y con solución salina en el masetero izquierdo (control intra-individuo). El volumen y grosor de esmalte y dentina del M1, y la longitud de la raíz mesial fueron medidos. Resultados: No hubo cambios en el volumen del esmalte del lado tratado con BoNTA y en ambos lados del grupo Control, 2 semanas post-intervención. El esmalte del lado control intra-individuo se redujo comparado con el lado experimental (p< 0,01). No hubo cambios en el volumen de dentina, el grosor de esmalte y dentina o en longitud de la raíz mesial de ambos grupos. Conclusión: La cantidad de esmalte en los molares hipofuncionales no se afecta después de la inyección unilateral de BoNTA en masetero, pero si se reduce en el lado contralateral. Por lo tanto, se debe considerar un desgaste dental asimétrico durante esta intervención.

Microbiological quality of honey from stingless bee, jandaíra (Melipona subnitida), from the semiarid region of Brazil / Qualidade microbiológica do mel da abelha sem ferrão jandaíra (Melipona subnitida) da região semi-árida do Brasil

ABSTRACT: The aim of this study was to evaluate the presence of microorganisms in honey produced by the stingless bee (SB) jandaíra (Melipona subnitida) from the semiarid region of Brazil. Thirty-five samples were analyzed and all of them were positive for mesophilic bacteria, coliforms at 45°C, fungi, and yeast. Staphylococcus spp. were identified in 85.7%, while Bacillus spp. were detected in 34.3% of honey samples. DNAs of Clostridium perfringens and C. botulinum were detected in 40% and 2.8% of the samples, respectively. Salmonella spp. and C. difficile were not detected. The present research revealed a great diversity of microorganisms in honey produced by jandaíra.

RESUMO: O objetivo deste trabalho foi avaliar a presença de microrganismos no mel produzido pela abelha sem ferrão jandaíra (Melipona subnitida). Trinta e cinco amostras foram avaliadas e todas foram positivas para bactérias mesofílicas, coliformes a 45 ºC, fungos e leveduras. Staphylococcus spp. foi identificado em 85,7% enquanto Bacillus foi detectado em 34,3% das amostras de mel. Clostridium perfringens e C. botulinum foram detectados em 40% e 2,8% das amostras respectivamente. Salmonella spp. e C. difficile não foram detectados. O presente trabalho revelou uma grande diversidade de microrganismos no mel produzido por jandaíra.

Clostridium botulinum spores in Polish honey samples

The aim of this study was an examination of 240 multifloral honey samples collected from Polish apiaries to determine Clostridium botulinum occurrence. Honey was collected from apiaries directly after the extraction process. Samples were inoculated by using the dilution and centrifugation method. Suspected isolates were examined by using mouse bioassay, polymerase chain reaction (PCR), and real-time PCR methods. C. botulinum type A and B strains were detected in 5 of 240 examined honey samples (2.1%). Bacterial strains were also detected that were phenotypically similar to C. botulinum but that did not exhibit the ability to produce botulinum toxins and did not show the presence of the botulinum cluster (ntnh and bont genes) or expression of the ntnh gene. The methods used in the examination, especially the expression analysis of ntnh gene, enabled specific analysis of suspected strains and could be used routinely in environmental isolate analyses of C. botulinum occurrence.

Surto de botulismo tipo C em equinos no Rio Grande do Sul / Outbreak of type C botulism in horses in Rio Grande do Sul

O botulismo nos animais domésticos é uma intoxicação fatal ocasionada pela ingestão da neurotoxina de Clostridium botulinum. Este trabalho descreve os aspectos epidemiológicos, clínicos e laboratoriais de um surto de botulismo tipo C em equinos de uma propriedade rural no município de Porto Alegre, Rio Grande do Sul. Do total de 20 equinos, sete (35%) foram acometidos e morreram, e um eutanasiado após apresentar prognóstico desfavorável. Os cavalos, criados extensivamente, apresentaram sinais clínicos semelhantes, caracterizados por dificuldade de locomoção, letargia, paralisia flácida progressiva, tremores musculares, sudorese, ataxia e quedas frequentes. Quatro equinos foram necropsiados e não apresentaram lesões macroscópicas e microscópicas significativas. Amostras de encéfalo e medula espinhal foram enviados para realização do exame de imunofluorescência direta e foram negativas para o vírus da raiva. Fragmentos de fígado dos quatro equinos necropsiados, amostras de sedimento do solo e água suspeita de dois açudes da propriedade foram avaliados pelo exame direto para a verificação da presença de toxina botulínica e/ou esporos de Clostridium botulinum. No bioensaio em camundongos e soroneutralização pode-se detectar toxina botulínica tipo C em uma amostra de fígado, ao passo que as amostras de sedimento do solo e água foram negativas em todos os ensaios. Com base nos dados epidemiológicos, clínicos e laboratoriais pode-se estabelecer o diagnóstico etiológico de botulismo causado pela toxina botulínica do tipo C, e sugere-se, a água estagnada dos açudes como aprovável fonte da toxina.(AU)

Botulism in domestic animals is a fatal poisoning caused by ingestion of Clostridium botulinum neurotoxin. This paper describes epidemiological, clinical and laboratorial aspects of a botulism type C outbreak in horses in a farm in Porto Alegre, Rio Grande do Sul. From a total of 20 horses, seven (35%) were affected and died, and one was euthanized after presenting poor prognosis. The horses, which were raised extensively, showed similar clinical signs, that included limited mobility, lethargy, progressive flaccid paralysis, muscle tremors, sweating, ataxia, and frequent falls. Four horses were necropsied and did not show significant macroscopic and microscopic lesions. Brain and spinal cord samples were analyzed by direct immunofluorescence and were negative for rabies. Liver fragments from the four necropsied horses, soil sediment samples, and suspect water from two reservoirs were evaluated by direct exam to verify the botulinum toxin and/or Clostridium botulinum spores presence. In the serum neutralization bioassay in mice, the type C botulinum toxin was detected in one liver sample, at the same time that the soil sediment and water samples were negative in all assays. Based on the epidemiological, clinical and laboratory data the etiological diagnosis of botulism was established by being caused by botulinum toxin type C, and it is suggested that the stagnant water of the dams is the likely source of the toxin.(AU)

Type C waterborne botulism outbreaks in buffaloes (Bubalus bubalis) in the Amazon region / Surtos de botulismo hídrico tipo C em búfalos (Bubalus bubalis) na região amazônica

Botulism is a poisoning caused by botulinum neurotoxins (BoNTs). BoNTs serotypes C and D are involved in botulism outbreaks in cattle in several countries. Despite the high number of buffaloes worldwide, the real impact of botulism in buffaloes is not known, because it is not a notifiable disease in Brazil and only few studies have evaluated the occurrence of the disease in buffaloes. Those studies did not conduct diagnostic tests to confirm the presence of BoNTs. The objective of the present study was to describe three outbreaks of botulism in buffaloes in the Brazilian Amazon region considering epidemiological and clinical data as well as laboratory diagnosis to confirm the presence of BoNTs. The results of the bioassay were negative in the tissues and in feed samples, but positive for BoNT C in water samples. Confirmation of the occurrence of botulism in buffaloes allows the implementation of preventive strategies in susceptible herds. Waterborne botulism in buffaloes is prevented by ensuring the constant circulation of water collections and restricting the presence of dead animals and bones in order to prevent the accumulation of organic matter and the development of anaerobic conditions, which might favor the replication of Clostridium botulinum. Another measure that can be adopted is the shading of the pasture, in order to maintain the thermal comfort for the buffaloes and to avoid the excess of permanence of them in the water pools.(AU)

Botulismo é uma intoxicação causada por neurotoxinas botulínicas (BoNTs). Os sorotipos C e D de BoNTs estão envolvidos em surtos de botulismo em bovídeos em vários países. Apesar do elevado número de búfalos em todo o mundo, o real impacto do botulismo em búfalos não é conhecido; pois não é uma doença de notificação obrigatória no Brasil e poucos estudos avaliaram a incidência desta doença em búfalos. Além disso, estes estudos não realizaram testes diagnósticos para confirmar a presença de BoNTs. O objetivo do presente estudo foi descrever três surtos de botulismo em búfalos na região amazônica brasileira, considerando dados epidemiológicos e clínicos, bem como o diagnóstico laboratorial para confirmar a presença de BoNTs. Os resultados do bioensaio em camundongos foram negativos em todos os tecidos e nas amostras de alimentos testados; no entanto foram positivos para BoNT C nas amostras de água. A confirmação da ocorrência de botulismo em búfalos permite a implementação de estratégias preventivas nos rebanhos. O botulismo hídrico nos búfalos pode ser prevenido assegurando-se que coleções de água fossem mantidas limpas, sem a presença de animais mortos e ossadas no seu interior e não permitindo o acúmulo de matéria orgânica e condições de anaerobiose favoráveis à multiplicação de Clostridium. botulinum. Outra medida que pode ser adotada é o sombreamento das pastagens, a fim de manter o conforto térmico dos búfalos e assim evitar o excesso de sua permanência dentro das fontes de água.(AU)

O uso da toxina botulínica na correção do sorriso gengival- revisão de literatura / Use of the botulinum toxin in correction of the gingival smile - Literature review

A toxina botulínica inicialmente era utilizada apenas para tratamentos terapêuticos, depois de alguns estudos sua aplicação passou a ser também utilizada para tratamentos estéticos. Esta substância é produzida pela bactéria Clostridium botulinum, responsável pelo botulismo, doença que provoca intoxicação por alimentos mal conservados. Utilizada na medicina para tratar inicialmente patologias, hoje seu uso passa a ser também estético na odontologia. Alguns pacientes se queixam do sorriso gengival causado por uma exposição excessiva da gengiva e buscam tratamentos mais rápidos e com menor morbidade, neste caso a toxina botulínica é muito eficaz para a correção do sorriso gengival diagnosticado por uma hiperfunção muscular. Quando causado por excesso vertical maxilar, extrusão ou erupção passiva alterada dos dentes ântero-superiores, o sorriso gengival pode ser tratado com a cirurgia ortognática, tratamento ortodôntico, miectomia, reposicionamento labial e/ou gengivoplastia. O uso desta substância na odontologia, pode ser mais uma opção para amenizar ou corrigir o sorriso gengival por hiperfunção muscular ou associar a outros tipos de tratamentos. Desta forma, o objetivo deste trabalho é fazer uma revisão literária sobre o uso da toxina botulínica na correção do sorriso gengival e descrever seu protocolo de uso (AU)

Botulinum toxin initially was used only for therapeutic treatments, after some studies its application was also used for aesthetic treatments. This substance was produced by the bacterium Clostridium botulinum, responsible for botulism, a disease that causes intoxication by poorly preserved foods. Used in medicine to treat initially pathologies, today its use is now also aesthetic in dentistry. Some patients complain of the gingival smile caused by excessive exposure of the gums and seek faster treatments with lower morbidity; in this case the botulinum toxin is very effective for the correction of the gingival smile diagnosed by a muscular hyperfunction. When caused by excessive vertical jaw, extrusion or passive eruption of the upper anterior teeth, the gingival smile may be treated with orthognathic surgery, orthodontic treatment, myectomy, lip repositioning and / or gingivoplasty. The use of this substance in dentistry may be another option to soften or correct the gingival smile due to muscular hyperfunction or to associate with other types of treatments. In this way, the objective of this work is to make a literary review on the use of botulinum toxin in the correction of the gingival smile and to describe its protocol of use. (AU)

Comparative study on PCR genotyping methods of Clostridium botulinum / 中国食品卫生杂志

Objective To compare four PCR genotyping method for Clostridium botulinum,and provide the reliable method for detection and identification of Clostridium botulinum from surveillance and foodborn poisoning in Sichuan Province.Methods Six strains of C.botulinum types A,B and E were used to compare four PCR genotyping method-one muhiplex PCR method was from US FDA,two multiplex PCR method and one real-time PCR method were from ISO,and the differences were preliminarily analyzed.Results Three multiplex PCR method could detect C.botulinum types A,B and E in a single reaction.The expected bands for type A were vague using ISO multiplex PCR method 1,whereas bright expected bands could be obtained in the identification of C.botulinum by the other two multiplex PCR method.Real-time multiplex PCR method could detect different types of C.botulinum simultaneously;however,classification should be carried out separately because fluorescent labels were the same.Conclusion Multiplex PCR method from FDA and multiplex PCR method 2 from ISO were relatively simple and could be recommended for C.botulinum surveillance in Sichuan Province.

Clinical analysis of 53 patients withClostridium botulinum food poisoning / 中华危重病急救医学

Objective To analyze the epidemiologic data of patients withClostridium botulinum food poisoning, and to improve the understanding, diagnosis and treatment of food borne botulism.Methods A retrospective study was conducted. Fifty-three patients withClostridium botulinum food poisoning admitted to Chinese PLA Center of Poisoning and Treatment from January 2009 to December 2016 were enrolled, and they were divided into mild, moderate, and severe groups according to the severity of disease. The clinical data including medical history, epidemiology data, routine blood test and blood biochemistry at hospital admission, the vital signs and arterial blood gas analysis before and after treatment, as well as the occurrence frequency of symptom and sign on set were collected.Results Fifty-three patients with food borne botulism were enrolled, with 33 patients in mild group, 13 in moderate group, and 7 in severe group. Most of the patients were female, the age distribution was in large span, the outbreak of disease was in groups mainly with the family, and patients were mainly located in Hebei Province, Beijing and Henan Province. The outbreaks were mainly happened in Spring and Summer, and homemade fermentation products were still the first cause of poisoning with the average latent period of (51.01±4.78) hours. The majority of patients with botulism were in mild resulted from the type A toxin. With the aggravation of disease, hospitalization time was gradually increased, white blood cell (WBC) and neutrophils (NEUT) at hospitalization admission, and respiratory rate (RR), heart rate (HR), fraction of inspired oxygen (FiO2) before the treatment were shown in obviously rising trend, albumin (ALB) at hospitalization admission and pH, arterial partial pressure of oxygen (PaO2), arterial oxygen saturation (SaO2) before treatment were in decline. The parameters in severe group were most severe, and had significant differences as compared with those of mild group [hospitalization time (days): 72.57±39.52 vs. 6.61±3.72, WBC (×109/L): 13.01±6.44 vs. 6.85±2.07, NEUT: 0.85±0.07 vs.0.63±0.14, RR (bpm): 32.14±4.33 vs. 15.18±1.70, HR (bpm): 132.29±5.19 vs. 75.54±8.24, FiO2: 0.32±0.05 vs. 0.21±0.00, ALB (g/L): 38.57±4.65 vs. 42.09±4.57, pH: 7.08±0.10 vs. 7.38±0.07, PaO2 (mmHg, 1 mmHg = 0.133 kPa): 75.16±5.24 vs. 98.39±1.50, SaO2: 0.78±0.06 vs. 0.97±0.02, allP < 0.05]. The symptom and sign on set of 53 patients with food borne botulism was dizziness, followed by fatigue, blurred vision, nausea, and other symptoms and signs were lower than 50%, and the occurrence of dizziness with rank one happen rate was significantly higher than blurred vision and nausea (χ2 values were 7.209 and 10.502 respectively, andP values were 0.007 and 0.004 respectively). After the on time prescription of botulinum antitoxins treatment, the clinical symptoms of patients could be relived quickly. All the patients were discharged without deaths.Conclusion In order to improve the recovery of the food borne botulism poisoning patients, adequate antitoxin and the related organ supports should be prescribed on time.

Molecular and Epidemiological Characterization of Infant Botulism in Beijing, China / 生物医学与环境科学（英文）

Laboratory-based pathogen isolation, identification, and toxicity determination were performed on samples from a suspected case of infant botulism. Mice injected with cultures generated from the enema sample and ingested Powered infant formula (PIF) presented typical signs of botulism. Antitoxins to polyvalent botulinum neurotoxins (BoNTs) and monovalent BoNT type B antitoxin had protective effects. Clostridium botulinum isolated from the enema and residual PIF samples were positive for type B toxin. Pulsed-field gel electrophoresis (PFGE) revealed that the two strains of C. botulinum isolated from the two samples produced indistinguishable pulsotypes. These findings confirmed this case of type B infant botulism associated with the ingestion of PIF contaminated by type B C. botulinum spores.

Botulism in non-ruminants in Brazil / Botulismo em não ruminantes no Brasil

ABSTRACT: Botulism is an intoxication caused by the ingestion of neurotoxins secreted by Clostridium botulinum and characterized by progressive flaccid symmetrical paralysis. Among non-ruminant animals, avian species and dogs are the most commonly affected by botulism, while horses and pigs are less-commonly diagnosed with the disease. Despite the importance of this disease in animals, the Brazilian literature only includes case studies and outbreak descriptions. The aim of this study was to review the main features and to provide recent data on the occurrence of botulism in non-ruminants in Brazil.

RESUMO: O botulismo é uma intoxicação causada pela ingestão das neurotoxinas secretadas por Clostridium botulinum e caracterizada por uma paralisia flácida simétrica progressiva. Entre os animais não ruminantes, aves e cães são os mais comumente afetados, enquanto equinos e suínos são esporadicamente diagnosticados com a doença. Apesar da reconhecida importância dessa doença nos animais, a literatura brasileira baseia-se apenas em relatos de casos e descrições de surtos. O presente trabalho tem como objetivo revisar a ocorrência e principais característicasdo botulismo em animais não ruminantes no Brasil.

Botulism in ruminants in Brazil / Botulismo em ruminantes no Brasil

ABSTRACT: Botulism is an intoxication caused by the ingestion of neurotoxins secreted by Clostridium botulinum. It affects all warm-blooded animals and some fishes and is characterized by progressive flaccid symmetrical paralysis. Despite the known importance of this disease in ruminants, the Brazilian literature only includes case studies and outbreak descriptions. In addition, the epidemiology of botulism in cattle has markedly changed over the past two decades. The aim of this study is to review these characteristics and to provide recent data on the occurrence of botulism in ruminants in Brazil.

RESUMO: O botulismo é uma intoxicação causada pela ingestão das neurotoxinas secretadas por Clostridium botulinum . A doença afeta todos os animais de sangue quente e alguns tipos de peixes, sendo caracterizada por uma paralisia flácida simétrica progressiva. Apesar da reconhecida importância dessa doença nos ruminantes, a literatura brasileira baseia-se apenas em relatos de casos e descrições de surtos. Além disso, a epidemiologia do botulismo em bovinos apresentou marcantes alterações nas últimas duas décadas. Dessa forma, é essencial uma revisão que elucide as principais características da doença em animais no Brasil. O presente trabalho tem como objetivo revisar tais características, além de fornecer dados recentes sobre a sua ocorrência em ruminantes.

The use of botulinum toxin as a way of treatment for gingival smile: case report

Gingival smile is defined as excessive gum exposition. Its etiology may be related to several factors, among them a maxillary vertical excess, superior dentoalveolar protrusion, changed passive extrusion and or eruption of anterior-superior teeth and hyperactivity of superior lip lift muscle. In cases in which the etiological factor is from muscle, the use of Botulinum toxin typo A (BTX-A) should be indicated. To highlight the benefits obtained with therapeutic use of this substance, the authors describe a clinical case of a female young patient, in which the Botox® was applied to correct gingival smile as a way to complement the orthodontic treatment. The applications were performed in two lateral points on the nose wing in a single session. The result was very satisfactory and there was no need additional dose. The authors concluded that BTX-A is a safe option and minimum invasive for treatment of gingival smile when provoked by muscle hypercontraction. Its side effects are rare with discrete discomfort during the application.

Invaginación intestinal en un lactante con diagnóstico de botulismo: Reporte de un caso / Intussusception in infant with diagnostic botulism: A case report

El botulismo es un importante problema de salud pública en Argentina. Es una enfermedad potencialmente letal y de difícil diagnóstico. Existen casos de presentación infrecuente de dicha enfermedad, como el abdomen agudo. Exponemos el caso de un niño de 4 meses, que consultó por constipación de 3 días de evolución, asociada a decaimiento y regular actitud alimentaria de 12 horas de evolución. Presentaba tono muscular conservado, sin alteraciones en la succión ni deglución, según la referencia materna. Se constató sensorio alternante y abdomen agudo, por lo que ingresó a quirófano con sospecha de invaginación intestinal, la cual fue confirmada mediante desinvaginación neumática. Durante la internación, el paciente evolucionó desfavorablemente y presentó llanto débil, hipotonía progresiva e insuficiencia respiratoria, por lo que requirió cuidados intensivos. Se aisló Clostridium botulinum en la muestra de materia fecal y toxina botulínica tipo A en el suero. Recibió toxina antibotulínica equina como tratamiento, con recuperación total a los 25 días de haber ingresado.

Botulism is an important public health problem in Argentina. It is a potentially fatal disease, and its diagnosis may be difficult. There are rare presentation forms of the disease, such as acute abdomen. We present a 4-monthbaby with a 3-day constipation condition, associated with weakness and abnormal eating attitude in the last 12 hours. The baby presented preserved muscle tone, with no changes in sucking or deglutition according to the mother's observations. Altered sensorium and acute abdomen were found; the patient was entered into the operating room with presumptive diagnosis ofintussusception, which was confirmed by pneumatic desinvagination. During hospitalization, the patient did not make good progress and presented weak cry, progressive hypotonia and respiratory failure requiring intensive care. Clostridium botulinum was isolated from the stool sample and botulinum toxin type A was isolated from serum. The patient was treated with equine botulinum toxin. Twenty five days after admission, he was totally recovered.

Estudo retrospectivo de surtos de botulismo no Brasil e no mundo

A distribuição do botulismo é mundial, com casos esporádicos ou surtos familiares, em geral, relacionados à produção e à conservação de alimentos de maneira inadequada. As intoxicações alimentares constituem um dos significantes problemas de saúde pública. O botulismo é uma doença grave, causada pelo Clotrisdium botulinum e deve ser considerado emergência médica e de saúde pública. De ocorrência súbita, caracteriza-se por manifestações neurológicas seletivas, de evolução dramática e elevada mortalidade, entre 30 e 65%. Devido a importância do botulismo como um problema de saúde pública, é indispensável relatar que o maior perigo de contaminação está nos alimentos preparados de forma artesanal, principalmente em conservas caseiras.

The distribution of botulism is worldwide, with sporadic cases or outbreaks family usually related to production and conservation of food improperly. The food poisoning is one of the significant public health problems. Botulism is a serious illness caused by Clostridium botulinum should be considered a medical emergency and public health. The sudden occurrence characterized by neurological manifestations selective evolution of dramatic and high mortality between 30 and 65%. Due to the importance of botulism as a public health problem, it is essential to report that the greatest danger of contamination in food is prepared by hand, especially in home canning.

Surto de botulismo tipo C em aves domésticas no semiárido do Nordeste, Brasil / Outbreak of type C botulism in poultry in the semiarid Northeast, Brazil

O presente estudo tem como objetivo relatar um surto de botulismo tipo C em galinhas (Gallus gallus domesticus), patos (Anas platyrhynchos) e em galinhas-d'-angola (Numida meleagris) criadas no semiárido do Rio Grande do Norte, Brasil. As aves incluídas neste estudo foram oriundas de um criatório com 80 animais, sendo 48 galinhas, 12 patos e 20 galinhas-d'angola. Os animais apresentaram paralisia flácida simétrica das patas, asas, pescoço e pálpebras, desprendimento de penas e dispneia. Vinte e quatro horas após início dos sinais clínicos constatou-se a mortalidade de 30 aves (37,5%). Após 48 horas, os 50 animais restantes também apresentaram sinais clínicos similares e vieram a óbito. Três aves com sinais clínicos de botulismo foram enviadas ao Laboratório de Patologia Veterinária da Universidade Federal Rural do Semi-Árido, onde coletou-se sangue e realizou-se necropsia seguida de avaliação anatomopatológica. Não foram observadas lesões macro e microscópicas nos órgãos da cavidade celomática e encéfalo. As alíquotas de soro foram encaminhadas para o Setor de Clostridioses do Laboratório Nacional Agropecuário de Minas Gerais (LANAGRO-MG) para pesquisa da toxina botulínica por soroneutralização em camundongos, que resultou na detecção da toxina botulínica tipo C. Este é o primeiro relato de botulismo em aves na região Nordeste do Brasil, e a primeira descrição da doença em galinhas d'Angola.

This study aims to report an outbreak of type C botulism in chickens (Gallus gallus domesticus), ducks (Anas platyrhynchos) and guinea fowls (Numida meleagris) created in semiarid region of Rio Grande do Norte, Brazil. The birds included in this study were derived from a farm with 80 animals (48 chickens, 12 ducks and 20 guinea fowls). The birds had symmetrical flaccid paralysis of the legs, wings, neck and eyelids, shedding feathers and dyspnea. After 24 hours of onset of clinical signs, 30 (37.5%) of the birds were found dead. After 48 hours, the 50 remaining animals also showed similar clinical signs and came to death. Three birds with clinical signs of botulism were sent to the Laboratory of Veterinary Pathology, of the Universidade Federal Rural do Semiárido, where blood was collected and held necropsy followed by histopathologic evaluation. There were no gross and microscopic lesions in the organs of the coelomic cavity and brain. There were no gross and microscopic lesions in the organs of the coelomic cavity and brain. The serum samples were sent to the Clostridial Section of the Laboratório Nacional Agropecuário de Minas Gerais (LANAGRO- MG) to search botulinic toxin by serumneutralization, which resulted in the detection of type C botulinic toxin. This is the first description of botulism in avian in the Northeastern Brazil and the first report of the disease in guinea fowls.