Type C botulism outbreak in free-ranging waterfowl in Goiás / Surto de botulismo tipo C em aves aquáticas de vida livre em Goiás

Botulism is generally a fatal disease caused by ingestion of neurotoxins produced by Clostridium botulinum. The present study describes the epidemiological, clinical, and laboratory aspects of a type C botulism outbreak in free-living aquatic birds residing in an urban park in Quirinópolis, Goiás, Brazil. Among a population of approximately 80 waterfowl, a total of 30 birds, including ducks (Cairina moschata), teals (Anas platyrhynchos), and geese (Anser cygnoides), died within 10 days. Of these, six birds showed signs of flaccid paralysis of the pelvic limbs, eyelids, neck, and wings. To confirm the suspicion of botulism, four lake water samples, two samples of the feed consumed by the birds, and samples of serum, intestinal content, stomach content, and liver tissue from two teals that died after presenting clinical signs were analyzed. Using bioassay and neutralization with homologous antitoxin in mice, it was possible to detect the presence of botulinum toxin type C in a water sample and in the intestinal content of one of the necropsied teals. Additionally, the presence of C. botulinum type C was identified in the lake water using polymerase chain reaction. Based on the clinical signs and laboratory results, a diagnosis of botulism caused by botulinum toxin type C was confirmed with probable transmission by lake water.

O botulismo é uma doença geralmente fatal, causada pela ingestão de neurotoxinas produzidas pelo Clostridium botulinum. O presente estudo descreve os aspectos epidemiológicos, clínicos e laboratoriais de um surto de botulismo tipo C em aves aquáticas de vida livre habitantes de parque urbano em Quirinópolis, Goiás. De uma população de cerca de 80 aves aquáticas, um total de 30 aves, entre patos (Cairina moschata), marrecos (Anas platyrhynchos) e gansos (Anser cygnoides), morreram no intervalo de 10 dias. Destes, seis aves apresentaram sinais de paralisia flácida de membros pélvicos, pálpebras, pescoço e asas. Para confirmar a suspeita de botulismo, foram analisadas quatro amostras da água do lago, duas amostras da ração consumida pelas aves e amostras de soro, conteúdo intestinal, conteúdo estomacal e fígado de dois marrecos que morreram após apresentarem os sinais clínicos. Pelo bioensaio e neutralização com antitoxina homóloga em camundongos foi possível detectar a presença de toxina botulínica tipo C em uma amostra de água e no conteúdo intestinal de um dos marrecos necropsiados. Adicionalmente, pela reação em cadeia da polimerase identificou-se a presença de C. botulinum tipo C na água do lago. Com base nos sinais clínicos e resultados laboratoriais estabeleceu-se o diagnóstico de botulismo causado pela toxina botulínica tipo C e veiculada provavelmente pela água do lago.

Botulinum toxin as a treatment for short bowel syndrome in rats

Purpose: The denervation of the intestine with benzalkonium chloride (BAC) reduces mortality and improves weight gain in rats with short bowel syndrome (SBS). Nevertheless, translating these promising findings from bench to bedside is not feasible because BAC promotes peritonitis and irreversible denervation which may be followed by an uncontrolled dilatation of the viscera. The use of botulinum toxin (BT) instead of BAC to achieve the denervation of the remaining small intestine in SBS could be an interesting option because it leads to a mild and transient denervation of the intestine. Methods: Here we evaluated the effects of the ileal denervation with BT in rats with SBS by verifying the body weight variation and intestinal morphological parameters. Four groups with 6 animals each were submitted to enterectomy with an ileal injection of saline (group E) or BT (group EBT). Control groups were submitted to simulated surgery with an ileal injection of BT (group BT) or saline (group C control). Results: We observed that the treatment of the remaining ileum with BT completely reversed the weight loss associated to extensive small bowel resection. Conclusion: This may provide a new promising approach to the surgical treatment of SBS.(AU)

Influência do uso da associação da xilazina e quetamina em Cavia porcellus, na potência de toxinas botulínicas na soro neutralização / Influence of the use of xylazine and ketamine association in Cavia porcellus on the potency of botulinum toxins and d in serum neutralization

O botulismo é uma zoonose causada pelas neurotoxinas produzidas por Clostridium botulinum. Existem oito variedades de toxinas (A-H), as toxinas C e D são as principais responsáveis por prejuízos na bovinocultura, causando óbito. Dentre as medidas preventivas eficientes, destaca-se a vacinação dos animais. Uma metodologia padrão para medir a eficiência de uma vacina é a soroneutralização in vitro tendo camundongos como sistema indicador. O objetivo do trabalho foi verificar a interferência do uso de anestésicos em Cavia porcellus vacinados sobre o efeito protetor dos soros, quando realizada a coleta de sangue para realizar a soroneutralização frente às toxinas padrões C e D. Neste trabalho foram realizados procedimentos de vacinação subcutânea, na região laterodorsal de Cavia porcellus, na dose de 5 ml no primeiro dia e repetição após 21 dias, e sangria após mais 21 dias. Foram formados dois grupos de 10 animais para cada amostra vacinal, um para sangria com o uso de associação anestésica (cloridrato de xilazina 2%, 5mg/Kg/IM e cloridrato de quetamina 10%, 25mg/Kg/IM) e outro sem uso, após isso submeteu-se o pool de soro coletado de cada grupo à soroneutralização in vitro com toxinas padrões C e D. Esses procedimentos de validação possibilitarão o seu emprego visando o bem-estar animal nos testes de controle de qualidade de vacinas. Foi observado que a aplicação de anestésicos para a punção cardíaca não interferiu no nível de proteção dos soros dos animais frente ao desafio de toxinas C e D, comparado aos não anestesiados.

Botulism is a zoonosis caused by neurotoxins produced by Clostridium botulinum. There are eight varieties of toxins (A-H), toxins C and D are main responsible for illness in cattle, causing death. Among the effective preventive measures, the vaccination of the animals stands out. To ensure adequate protection of animals, vaccines must be efficient. A standard methodology for measuring the efficiency of a vaccine is in vitro serum neutralization having mice as the indicator system. The objective of the study was to verify the interference of anesthetic use in vaccinated guinea pigs on the protective effect of sera when blood collection was performed to perform serum neutralization against standard C and D toxins. In this work, subcutaneous vaccination procedures were performed in the later dorsal region of guinea pigs at the dose of 5 ml on the first day and repetition after 21 days and bleeding after another 21 days. Two groups of 10 animals were formed for each sample de vaccine, one for bleeding with the use of an anesthetic association (xylazine hydrochloride 2%, 5mg/Kg/IM and ketamine hydrochloride 10%, 25mg/Kg/IM) and the other with no use, after which the serum mixture collected from each group were submitted to in vitro serum neutralization with standard C and D toxins. This validation study will make it possible to introduce the use this procedures aimed at animal welfare in the quality control tests of vaccines. It was observed that the application of anesthetics to cardiac puncture did not interfere in the level of protection of the sera of the animals against the challenge of toxins C and D compared to those obtained of not anesthetized animals.

Influência do uso da associação da xilazina e quetamina em Cavia porcellus, na potência de toxinas botulínicas na soro neutralização / Influence of the use of xylazine and ketamine association in Cavia porcellus on the potency of botulinum toxins and d in serum neutralization

O botulismo é uma zoonose causada pelas neurotoxinas produzidas por Clostridium botulinum. Existem oito variedades de toxinas (A-H), as toxinas C e D são as principais responsáveis por prejuízos na bovinocultura, causando óbito. Dentre as medidas preventivas eficientes, destaca-se a vacinação dos animais. Uma metodologia padrão para medir a eficiência de uma vacina é a soroneutralização in vitro tendo camundongos como sistema indicador. O objetivo do trabalho foi verificar a interferência do uso de anestésicos em Cavia porcellus vacinados sobre o efeito protetor dos soros, quando realizada a coleta de sangue para realizar a soroneutralização frente às toxinas padrões C e D. Neste trabalho foram realizados procedimentos de vacinação subcutânea, na região laterodorsal de Cavia porcellus, na dose de 5 ml no primeiro dia e repetição após 21 dias, e sangria após mais 21 dias. Foram formados dois grupos de 10 animais para cada amostra vacinal, um para sangria com o uso de associação anestésica (cloridrato de xilazina 2%, 5mg/Kg/IM e cloridrato de quetamina 10%, 25mg/Kg/IM) e outro sem uso, após isso submeteu-se o pool de soro coletado de cada grupo à soroneutralização in vitro com toxinas padrões C e D. Esses procedimentos de validação possibilitarão o seu emprego visando o bem-estar animal nos testes de controle de qualidade de vacinas. Foi observado que a aplicação de anestésicos para a punção cardíaca não interferiu no nível de proteção dos soros dos animais frente ao desafio de toxinas C e D, comparado aos não anestesiados.(AU)

Botulism is a zoonosis caused by neurotoxins produced by Clostridium botulinum. There are eight varieties of toxins (A-H), toxins C and D are main responsible for illness in cattle, causing death. Among the effective preventive measures, the vaccination of the animals stands out. To ensure adequate protection of animals, vaccines must be efficient. A standard methodology for measuring the efficiency of a vaccine is in vitro serum neutralization having mice as the indicator system. The objective of the study was to verify the interference of anesthetic use in vaccinated guinea pigs on the protective effect of sera when blood collection was performed to perform serum neutralization against standard C and D toxins. In this work, subcutaneous vaccination procedures were performed in the later dorsal region of guinea pigs at the dose of 5 ml on the first day and repetition after 21 days and bleeding after another 21 days. Two groups of 10 animals were formed for each sample de vaccine, one for bleeding with the use of an anesthetic association (xylazine hydrochloride 2%, 5mg/Kg/IM and ketamine hydrochloride 10%, 25mg/Kg/IM) and the other with no use, after which the serum mixture collected from each group were submitted to in vitro serum neutralization with standard C and D toxins. This validation study will make it possible to introduce the use this procedures aimed at animal welfare in the quality control tests of vaccines. It was observed that the application of anesthetics to cardiac puncture did not interfere in the level of protection of the sera of the animals against the challenge of toxins C and D compared to those obtained of not anesthetized animals.(AU)

Histological and immunohistochemical findings of the action of botulinum toxin in salivary gland: systematic review / Achados histológicos e imunohistoquímicos da ação da toxina botulínica em glândula salivar: revisão sistemática

The treatment of sialorrhea is necessary for the constant risks posed by hypersalivation. A new therapeutic option comes up with the application of botulinum toxin in salivary glands. However, little is known about its mechanism of action in glandular tissue. Based on the above, this work had the objective to systematically review the literature about the action of botulinum toxin on submandibular and parotid salivary glands tissues. Electronic search was performed in databases of great relevance for this study (PubMed, SciELO, HighWire, Crossref, Scopus, Science Direct, MEDLINE, OLDMEDLINE, Serials Database, NLM Catalog, LILACS and IBECS). Inclusion and exclusion criteria for articles were established, and a total number of 14 articles were selected and used. There are few publications that clarify how the salivary gland acini behave with application of botulinum toxin. Although, the immunohistochemical findings were consistent among authors, showing weak immunoreactivity in glands treated with botulinum toxin. Histometric data are divergent, requiring more detailed studies to answer the questions about the efficacy and safety of botulinum toxin in salivary glands.(AU)

O tratamento da sialorreia se faz necessário pelos constantes riscos trazidos por este estado de hipersalivação. Uma nova opção terapêutica surge com a aplicação da toxina botulínica em glândulas salivares. Entretanto, pouco se sabe sobre o seu mecanismo de ação no tecido glandular. Com base no exposto, este trabalho teve o objetivo de revisar sistematicamente na literatura a ação da toxina botulínica sobre o tecido das glândulas salivares submandibular e parótida. Foi realizada uma busca eletrônica em bases de dados de grande relevância para este estudo (PubMed, SciELO, HighWire, Crossref, Scopus, Science Direct, MEDLINE, OLDMEDLINE, Serials Database, NLM Catalog, LILACS e IBECS). Foram estabelecidos critérios de inclusão e exclusão para os artigos, e um “n” de 14 trabalhos foram selecionados e utilizados. São poucas as publicações que esclarecem como os ácinos das glândulas salivares se comportam mediante aplicação da toxina botulínica. Apesar de os achados imunohistoquímicos entre os autores serem concordantes, com imunorreatividade mais fracas nas glândulas tratadas com a toxina botulínica, os dados histométricos são divergentes e há questionamentos metodológicos, necessitando de mais estudos pormenorizados para responder as questões sobre a eficácia e segurança da toxina botulínica nas glândulas salivares.(AU)

The positive effect of Botulinum toxin type A on the viability of random flap in tobacco exposed in rats

PURPOSE:To evaluate the effect of Botulinum Toxin A in different time of tobacco exposure.METHODS:60 male, Wistar rats were divided into two tobacco exposure groups: a 2- month or a 4-month regimen. After this period, these two groups were subdivided as two: saline solution(SS) or botulinum toxin A(Bonta), at the time of the surgery. Seven days before the SS or Bonta injection, the animals were submitted to a random flap (3x10cm). On the seventh postoperative day, all animals were assessed for total flap area, viable area, and the viable/ total area ratio.RESULTS:This study showed a difference between groups 2-month saline vs. BontA injection (p=0.04); groups 4-month saline vs. BontA injection (p=0.001); groups 2-month saline vs. 4-month BontA (p=0.003), and, between groups 2- month BontA vs. 4-month saline(p=0.03).CONCLUSIONS:Botulinum Toxin A increased random flap viability in tobacco-exposed rats. Two months of tobacco exposure had the same effect as exposure for four months.(AU)

Botulinum toxin type A on cutaneous flap viability in diabetic and tobacco-exposed rats

To investigate the effect of Botulinum toxin A (BoNTA) on skin flap viability in healthy, tobacco-exposed and diabetic rats. Ninety male Wistar rats (250-300g) were randomly divided into six groups: control+saline (C1), control+BoNTA (C2), tobacco-exposed+saline (T1), tobacco-exposed+BoNTA (T2) diabetes+saline (D1) and diabetes+BoNTA (D2). A dorsal cutaneous flap (3×10cm) was performed. Survival area and total area of the flaps were measured. Lumen diameter, external arterial diameter and lumen/wall thickness ratio were recorded. Survival area increased in control group with BoNTA injection compared with control animals injected with saline (C2 x C1; 0.9±0.1 vs0.67±0.15, p= 0.001). A similar result was found in diabetes group injected with BontA (D2 x D1; 0.97±0.2 vs0.61±0.24, p=0.018). No difference was observed in skin flap viability in tobacco-exposed groups (T2 x T1; 0.74±0.24 vs 0.64±0.21, p=0.871). Lumen diameter (p= 0.004), external arterial diameter (p = 0.0046,) and lumen/wall thickness ratio (p= 0.003) were increased in diabetes+BoNTA-treated animals. This effect was not observed in control or in tobacco-exposed groups. Botulinum toxin A increased skin flap viability in control and diabetic rats on the seventh post-operative day. Increased lumen diameter, external arterial diameter, and lumen/wall thickness ratio were observed in the diabetes+BoNTA group. BoNTA had no effect in the tobacco-exposed group on the seventh postoperative day.(AU)

Sensibilidade toxicológica e especificidade do teste de microfixação de complemento na detecção de toxinas botulínicas C e D em meio de cultura e fígado de camundongos

The toxicological sensitivity and specificity of the micro-complement fixation test (MCF) for the detection of botulinum toxins C and D were studied in supernatants of the bacterial cultures and in livers of mice inoculated with lethal and sublethal doses. Botulinum toxins C and D were produced in Hemoline culture medium, titered through the determination of LD50 by the mouse test and adjusted to dilutions of 10, 1, 0.1, 0.01 and 0.001 LD50. Two experimental models were used to determine the toxicological sensitivity of MCF in the supernatant of the culture medium with the dilutions described, and also in liver extracts of mice weighing 20 g and inoculated with the same dilutions. Detection of the botulinum toxins was attempted in liver extracts of mice which had received lethal doses of the respective toxins, and in others which had been inoculated with sublethal doses and were sacrificed in intervals of 5 days. The results show that the toxicological sensitivity of MCF, regarding the two types of toxins at the level of 0.001 LD50, was 100% when the supernatants of the culture medium were tested; this means that the sensitivity was 100 times higher than with the mouse test. The toxicological sensitivity of MCF in the liver extracts of mice inoculated with 1 and 10 LD50 of botulinum toxins C and D was inferior, giving values of 100, 80, 89 and 72% respectively. By this test it was also possible to detect botulinum toxins type C and D in liver extracts of mice inoculated with sublethal doses, up to 15 days after the injection. The specificity of MCF was 88% and 92%, when liver extracts of healthy control mice were tested and when challenged with antitoxins C and D; and 100% when challenged with the supernatant of the culture medium. These results indicate that MCF could be of importance for research and could substitute in vivo tests.

No presente estudo pretendeu-se verificar a sensibilidade toxicológica e especificidade do Teste de Microfixação de Complemento (MCF) na detecção de toxinas botulínicas C e D no sobrenadante de cultivos bacterianos e em fígados de camundongos inoculados com doses letais e subletais. As toxinas foram produzidas em meio de cultura Hemoline, tituladas através da determinação da DL50 pelo Bioensaio em Camundongo e diluídas nas concentrações de 10, 1, 0,1, 0,01 e 0,001 DL50. Desta forma, foram utilizadas em dois modelos experimentais, onde foi determinada a sensibilidade toxicológica do MCF no sobrenadante do meio de cultura com as diluições descritas acima e ainda em extratos hepáticos de camundongos com peso corporal de 20g, inoculados com as mesmas diluições. A tentativa de evidenciação das toxinas botulínicas nos extratos hepáticos de camundongos foi realizada através da sua extração após a morte pela administração das doses letais e ainda pelo sacrifício dos animais inoculados com doses subletais, em intervalos de 5 dias. Os resultados evidenciaram uma sensibilidade toxicológica para o MCF de 100% para os dois tipos de toxinas ao nível de 0,01 DL50, quando testados os sobrenadantes de meio de cultura, portanto 100 vezes superior ao Bioensaio em Camundongo. A sensibilidade toxicológica do MCF, quando examinados extratos hepáticos de camundongos inoculados com 1 e 10 DL50 de toxinas botulínicas C e D, foi inferior, com valores de 100, 80, 89 e 72%, respectivamente. Pelo teste foi possível detectar toxinas botulínicas tipos C e D nos extratos hepáticos de camundongos inoculados com doses subletais até 15 dias após a sua inoculação. A especificidade do MCF foi de 88 e 92%, quando testados extratos hepáticos de camundongos sadios, e confrontados com as antitoxinas C e D; e 100% no sobrenadante do meio de cultura. Os resultados apontam para uma possível utilização do teste como importante instrumento de pesquisa e ainda na eventual substituição dos testes in vivo pelas suas implicações éticas e limitações práticas.