Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans

Abstract The study was aimed to assess impact of high fat diet (HFD) and synthetic human gut microbiota (GM) combined with HFD and chow diet (CD) in inducing type-2 diabetes (T2D) using mice model. To our knowledge, this is the first study using selected human GM transplantation via culture based method coupled dietary modulation in mice for in vivo establishment of inflammation leading to T2D and gut dysbiosis. Twenty bacteria (T2D1-T2D20) from stool samples of confirmed T2D subjects were found to be morphologically different and subjected to purification on different media both aerobically and anerobically, which revealed seven bacteria more common among 20 isolates on the basis of biochemical characterization. On the basis of 16S rRNA gene sequencing, these seven isolates were identified as Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenes (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). The seven isolates were subsequently used as synthetic gut microbiome (GM) for their role in inducing T2D in mice. Inbred strains of albino mice were divided into four groups and were fed with CD, HFD, GM+HFD and GM+CD. Mice receiving HFD and GM+modified diet (CD/HFD) showed highly significant (P 0.05) increase in weight and blood glucose concentration as well as elevated level of inflammatory cytokines (TNF-, IL-6, and MCP-1) compared to mice receiving CD only. The 16S rRNA gene sequencing of 11 fecal bacteria obtained from three randomly selected animals from each group revealed gut dysbiosis in animals receiving GM. Bacterial strains including Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) and Lactobacillus gasseri (MT152635) were isolated from mice treated with GM+modified diet (HFD/CD) compared to strains Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629) which were isolated from mice receiving CD/HFD. In conclusion, these findings suggest that constitution of GM and diet plays significant role in inflammation leading to onset or/and possibly progression of T2D. .

Resumo O estudo teve como objetivo avaliar o impacto da dieta rica em gordura (HFD) e da microbiota intestinal humana sintética (GM) combinada com HFD e dieta alimentar (CD) na indução de diabetes tipo 2 (T2D) usando modelo de camundongos. Para nosso conhecimento, este é o primeiro estudo usando transplante de GM humano selecionado através do método baseado em cultura acoplada à modulação dietética em camundongos para o estabelecimento in vivo de inflamação que leva a T2D e disbiose intestinal. Vinte bactérias (T2D1-T2D20) de amostras de fezes de indivíduos T2D confirmados verificaram ser morfologicamente diferentes e foram submetidas à purificação em meios diferentes aerobicamente e anaerobicamente, o que revelou sete bactérias mais comuns entre 20 isolados com base na caracterização bioquímica. Com base no sequenciamento do gene 16S rRNA, esses sete isolados foram identificados como Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenides (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). Esses sete isolados foram, posteriormente, usados como microbioma intestinal sintético (GM) por seu papel na indução de T2D em camundongos. Linhagens consanguíneas de camundongos albinos foram divididas em quatro grupos e foram alimentadas com CD, HFD, GM + HFD e GM + CD. Camundongos que receberam a dieta modificada com HFD e GM + (CD / HFD) mostraram um aumento altamente significativo (P 0,05) no peso e na concentração de glicose no sangue, bem como um nível elevado de citocinas inflamatórias (TNF-, IL-6 e MCP-1) em comparação com os ratos que receberam apenas CD. O sequenciamento do gene 16S rRNA de 11 bactérias fecais obtidas de três animais selecionados aleatoriamente de cada grupo revelou disbiose intestinal em animais que receberam GM. Cepas bacterianas, incluindo Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) e Lactobacillus Gasseri (MT152635D), foram tratadas com dieta modificada / CD) em comparação com as linhagens Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629), que foram isoladas de camundongos recebendo CD / HFD. Em conclusão, esses resultados sugerem que a constituição de GM e dieta desempenham papel significativo na inflamação levando ao início ou/e possivelmente à progressão de T2D.

Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans / Disbiose intestinal, inflamação e diabetes tipo 2 em camundongos usando microbiota intestinal sintética de humanos diabéticos

The study was aimed to assess impact of high fat diet (HFD) and synthetic human gut microbiota (GM) combined with HFD and chow diet (CD) in inducing type-2 diabetes (T2D) using mice model. To our knowledge, this is the first study using selected human GM transplantation via culture based method coupled dietary modulation in mice for in vivo establishment of inflammation leading to T2D and gut dysbiosis. Twenty bacteria (T2D1-T2D20) from stool samples of confirmed T2D subjects were found to be morphologically different and subjected to purification on different media both aerobically and anerobically, which revealed seven bacteria more common among 20 isolates on the basis of biochemical characterization. On the basis of 16S rRNA gene sequencing, these seven isolates were identified as Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenes (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). The seven isolates were subsequently used as synthetic gut microbiome (GM) for their role in inducing T2D in mice. Inbred strains of albino mice were divided into four groups and were fed with CD, HFD, GM+HFD and GM+CD. Mice receiving HFD and GM+modified diet (CD/HFD) showed highly significant (P<0.05) increase in weight and blood glucose concentration as well as elevated level of inflammatory cytokines (TNF-α, IL-6, and MCP-1) compared to mice receiving CD only. The 16S rRNA gene sequencing of 11 fecal bacteria obtained from three randomly selected animals from each group revealed gut dysbiosis in animals receiving GM. Bacterial strains including Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) and Lactobacillus gasseri (MT152635) were isolated from mice [...].

O estudo teve como objetivo avaliar o impacto da dieta rica em gordura (HFD) e da microbiota intestinal humana sintética (GM) combinada com HFD e dieta alimentar (CD) na indução de diabetes tipo 2 (T2D) usando modelo de camundongos. Para nosso conhecimento, este é o primeiro estudo usando transplante de GM humano selecionado através do método baseado em cultura acoplada à modulação dietética em camundongos para o estabelecimento in vivo de inflamação que leva a T2D e disbiose intestinal. Vinte bactérias (T2D1-T2D20) de amostras de fezes de indivíduos T2D confirmados verificaram ser morfologicamente diferentes e foram submetidas à purificação em meios diferentes aerobicamente e anaerobicamente, o que revelou sete bactérias mais comuns entre 20 isolados com base na caracterização bioquímica. Com base no sequenciamento do gene 16S rRNA, esses sete isolados foram identificados como Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenides (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). Esses sete isolados foram, posteriormente, usados como microbioma intestinal sintético (GM) por seu papel na indução de T2D em camundongos. Linhagens consanguíneas de camundongos albinos foram divididas em quatro grupos e foram alimentadas com CD, HFD, GM + HFD e GM + CD. Camundongos que receberam a dieta modificada com HFD e GM + (CD / HFD) mostraram um aumento altamente significativo (P < 0,05) no peso e na concentração de glicose no sangue, bem como um nível elevado de citocinas inflamatórias (TNF-α, IL-6 e MCP-1) em comparação com os ratos que receberam apenas CD. O sequenciamento do gene 16S rRNA de 11 bactérias fecais obtidas de três animais selecionados aleatoriamente de cada grupo revelou disbiose intestinal em animais que receberam GM. Cepas bacterianas, incluindo Bacteroides gallinarum (MT152630), Ruminococcus [...].

Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans / Disbiose intestinal, inflamação e diabetes tipo 2 em camundongos usando microbiota intestinal sintética de humanos diabéticos

Abstract The study was aimed to assess impact of high fat diet (HFD) and synthetic human gut microbiota (GM) combined with HFD and chow diet (CD) in inducing type-2 diabetes (T2D) using mice model. To our knowledge, this is the first study using selected human GM transplantation via culture based method coupled dietary modulation in mice for in vivo establishment of inflammation leading to T2D and gut dysbiosis. Twenty bacteria (T2D1-T2D20) from stool samples of confirmed T2D subjects were found to be morphologically different and subjected to purification on different media both aerobically and anerobically, which revealed seven bacteria more common among 20 isolates on the basis of biochemical characterization. On the basis of 16S rRNA gene sequencing, these seven isolates were identified as Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenes (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). The seven isolates were subsequently used as synthetic gut microbiome (GM) for their role in inducing T2D in mice. Inbred strains of albino mice were divided into four groups and were fed with CD, HFD, GM+HFD and GM+CD. Mice receiving HFD and GM+modified diet (CD/HFD) showed highly significant (P<0.05) increase in weight and blood glucose concentration as well as elevated level of inflammatory cytokines (TNF-α, IL-6, and MCP-1) compared to mice receiving CD only. The 16S rRNA gene sequencing of 11 fecal bacteria obtained from three randomly selected animals from each group revealed gut dysbiosis in animals receiving GM. Bacterial strains including Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) and Lactobacillus gasseri (MT152635) were isolated from mice treated with GM+modified diet (HFD/CD) compared to strains Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629) which were isolated from mice receiving CD/HFD. In conclusion, these findings suggest that constitution of GM and diet plays significant role in inflammation leading to onset or/and possibly progression of T2D. .

Resumo O estudo teve como objetivo avaliar o impacto da dieta rica em gordura (HFD) e da microbiota intestinal humana sintética (GM) combinada com HFD e dieta alimentar (CD) na indução de diabetes tipo 2 (T2D) usando modelo de camundongos. Para nosso conhecimento, este é o primeiro estudo usando transplante de GM humano selecionado através do método baseado em cultura acoplada à modulação dietética em camundongos para o estabelecimento in vivo de inflamação que leva a T2D e disbiose intestinal. Vinte bactérias (T2D1-T2D20) de amostras de fezes de indivíduos T2D confirmados verificaram ser morfologicamente diferentes e foram submetidas à purificação em meios diferentes aerobicamente e anaerobicamente, o que revelou sete bactérias mais comuns entre 20 isolados com base na caracterização bioquímica. Com base no sequenciamento do gene 16S rRNA, esses sete isolados foram identificados como Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenides (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). Esses sete isolados foram, posteriormente, usados ​​como microbioma intestinal sintético (GM) por seu papel na indução de T2D em camundongos. Linhagens consanguíneas de camundongos albinos foram divididas em quatro grupos e foram alimentadas com CD, HFD, GM + HFD e GM + CD. Camundongos que receberam a dieta modificada com HFD e GM + (CD / HFD) mostraram um aumento altamente significativo (P < 0,05) no peso e na concentração de glicose no sangue, bem como um nível elevado de citocinas inflamatórias (TNF-α, IL-6 e MCP-1) em comparação com os ratos que receberam apenas CD. O sequenciamento do gene 16S rRNA de 11 bactérias fecais obtidas de três animais selecionados aleatoriamente de cada grupo revelou disbiose intestinal em animais que receberam GM. Cepas bacterianas, incluindo Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) e Lactobacillus Gasseri (MT152635D), foram tratadas com dieta modificada / CD) em comparação com as linhagens Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629), que foram isoladas de camundongos recebendo CD / HFD. Em conclusão, esses resultados sugerem que a constituição de GM e dieta desempenham papel significativo na inflamação levando ao início ou/e possivelmente à progressão de T2D.

Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans / Disbiose intestinal, inflamação e diabetes tipo 2 em camundongos usando microbiota intestinal sintética de humanos diabéticos

The study was aimed to assess impact of high fat diet (HFD) and synthetic human gut microbiota (GM) combined with HFD and chow diet (CD) in inducing type-2 diabetes (T2D) using mice model. To our knowledge, this is the first study using selected human GM transplantation via culture based method coupled dietary modulation in mice for in vivo establishment of inflammation leading to T2D and gut dysbiosis. Twenty bacteria (T2D1-T2D20) from stool samples of confirmed T2D subjects were found to be morphologically different and subjected to purification on different media both aerobically and anerobically, which revealed seven bacteria more common among 20 isolates on the basis of biochemical characterization. On the basis of 16S rRNA gene sequencing, these seven isolates were identified as Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenes (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). The seven isolates were subsequently used as synthetic gut microbiome (GM) for their role in inducing T2D in mice. Inbred strains of albino mice were divided into four groups and were fed with CD, HFD, GM+HFD and GM+CD. Mice receiving HFD and GM+modified diet (CD/HFD) showed highly significant (P<0.05) increase in weight and blood glucose concentration as well as elevated level of inflammatory cytokines (TNF-α, IL-6, and MCP-1) compared to mice receiving CD only. The 16S rRNA gene sequencing of 11 fecal bacteria obtained from three randomly selected animals from each group revealed gut dysbiosis in animals receiving GM. Bacterial strains including Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) and Lactobacillus gasseri (MT152635) were isolated from mice [...].(AU)

O estudo teve como objetivo avaliar o impacto da dieta rica em gordura (HFD) e da microbiota intestinal humana sintética (GM) combinada com HFD e dieta alimentar (CD) na indução de diabetes tipo 2 (T2D) usando modelo de camundongos. Para nosso conhecimento, este é o primeiro estudo usando transplante de GM humano selecionado através do método baseado em cultura acoplada à modulação dietética em camundongos para o estabelecimento in vivo de inflamação que leva a T2D e disbiose intestinal. Vinte bactérias (T2D1-T2D20) de amostras de fezes de indivíduos T2D confirmados verificaram ser morfologicamente diferentes e foram submetidas à purificação em meios diferentes aerobicamente e anaerobicamente, o que revelou sete bactérias mais comuns entre 20 isolados com base na caracterização bioquímica. Com base no sequenciamento do gene 16S rRNA, esses sete isolados foram identificados como Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenides (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). Esses sete isolados foram, posteriormente, usados como microbioma intestinal sintético (GM) por seu papel na indução de T2D em camundongos. Linhagens consanguíneas de camundongos albinos foram divididas em quatro grupos e foram alimentadas com CD, HFD, GM + HFD e GM + CD. Camundongos que receberam a dieta modificada com HFD e GM + (CD / HFD) mostraram um aumento altamente significativo (P < 0,05) no peso e na concentração de glicose no sangue, bem como um nível elevado de citocinas inflamatórias (TNF-α, IL-6 e MCP-1) em comparação com os ratos que receberam apenas CD. O sequenciamento do gene 16S rRNA de 11 bactérias fecais obtidas de três animais selecionados aleatoriamente de cada grupo revelou disbiose intestinal em animais que receberam GM. Cepas bacterianas, incluindo Bacteroides gallinarum (MT152630), Ruminococcus [...].(AU)

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.

Surto de botulismo tipo C em aves de subsistência em Santa Catarina, Brasil / Outbreak of type C botulism in backyard poultry in Santa Catarina, Brazil

Background: Botulism is a disease caused by the ingestion of neurotoxin produced by Clostridium botulinum, characterizedby flaccid paralysis, which can lead to high mortality. They have seven types of neurotoxins (A, B, C, D, E, F, and G) and,in birds, most cases are attributed to type C. They are considered sources of botulinum toxins where the decomposition oforganic matter occurs, like stagnant water and rotting food. The main feature of the disease in birds is ascending symmetricflaccid paralysis. The present study aims to describe an outbreak of type C botulism in backyard poultry in the state ofSanta Catarina, Southern Brazil.Case: A visit was made to the property with 160 backyard poultry with a history of high mortality in the municipality ofAgrolândia, Santa Catarina. Clinical signs were characterized by paralysis of the pelvic limbs, neck and pendular wings,which progressed to death within 48 h. There was a mortality rate of 37.5% (60/160) between March and May 2019. Thesebirds were kept in an overcrowded environment, with different species (chickens, ducks, teals, and turkeys) fed irregularly.The water supplied was provided from kitchen exhaust, accumulating in puddles on the floor that contained organic matterresidues such as animal feces, food waste and bone fragments. The disposal of the carcasses of birds that died was in thesame enclosure, buried superficially, facilitating the access of other birds to dig them up and consume them. Necropsywas performed on 2 chickens and one duck, no macroscopic or histopathological lesions were observed. Blood, liver, andgastrointestinal content samples were sent for research and identification of botulinum toxin through the serum neutralization test in mice. The presence of type C botulinum toxin was confirmed in the liver chicken of one sampled animals.Discussion: The identification of type C botulism toxin enabled the characterization of the outbreak, which is...

Surto de botulismo tipo C em aves de subsistência em Santa Catarina, Brasil / Outbreak of type C botulism in backyard poultry in Santa Catarina, Brazil

Background: Botulism is a disease caused by the ingestion of neurotoxin produced by Clostridium botulinum, characterizedby flaccid paralysis, which can lead to high mortality. They have seven types of neurotoxins (A, B, C, D, E, F, and G) and,in birds, most cases are attributed to type C. They are considered sources of botulinum toxins where the decomposition oforganic matter occurs, like stagnant water and rotting food. The main feature of the disease in birds is ascending symmetricflaccid paralysis. The present study aims to describe an outbreak of type C botulism in backyard poultry in the state ofSanta Catarina, Southern Brazil.Case: A visit was made to the property with 160 backyard poultry with a history of high mortality in the municipality ofAgrolândia, Santa Catarina. Clinical signs were characterized by paralysis of the pelvic limbs, neck and pendular wings,which progressed to death within 48 h. There was a mortality rate of 37.5% (60/160) between March and May 2019. Thesebirds were kept in an overcrowded environment, with different species (chickens, ducks, teals, and turkeys) fed irregularly.The water supplied was provided from kitchen exhaust, accumulating in puddles on the floor that contained organic matterresidues such as animal feces, food waste and bone fragments. The disposal of the carcasses of birds that died was in thesame enclosure, buried superficially, facilitating the access of other birds to dig them up and consume them. Necropsywas performed on 2 chickens and one duck, no macroscopic or histopathological lesions were observed. Blood, liver, andgastrointestinal content samples were sent for research and identification of botulinum toxin through the serum neutralization test in mice. The presence of type C botulinum toxin was confirmed in the liver chicken of one sampled animals.Discussion: The identification of type C botulism toxin enabled the characterization of the outbreak, which is...(AU)

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.

Causes of death in feedlot beef cattle and their control: a brief review / Causas de morte em bovinos de corte confinados e seu controle: uma breve revisão

This review reports the leading causes of death in feedlot beef cattle. It describes economic losses resulting from these deaths and suggests control alternatives. Diseases associated with the respiratory and digestive systems were the most frequently observed. In different geographical areas, the importance of each one might vary. Outbreaks of diseases such as botulism occur occasionally and can cause important economic losses. Cattle tick fever can cause significant losses in zones of enzootic tick instability. Technical assistance and sanitary and food management are critical for the best productivity in feedlot cattle.(AU)

Esta revisão discute as principais causas de morte em bovinos de corte em confinamento. Descreve as perdas econômicas resultantes dessas mortes e sugere alternativas de controle. As doenças associadas aos sistemas respiratório e digestivo foram as mais frequentemente observadas. Em diferentes áreas geográficas, a importância de cada uma pode variar. Surtos de doenças como o botulismo ocorrem ocasionalmente e podem causar importantes perdas econômicas. A tristeza parasitária bovina pode causar perdas significativas em zonas de instabilidade enzoótica do carrapato. A assistência técnica e um bom gerenciamento sanitário e alimentar são essenciais para a melhor produtividade em bovinos de corte confinados.(AU)

Causes of death in feedlot beef cattle and their control: a brief review / Causas de morte em bovinos de corte confinados e seu controle: uma breve revisão

This review reports the leading causes of death in feedlot beef cattle. It describes economic losses resulting from these deaths and suggests control alternatives. Diseases associated with the respiratory and digestive systems were the most frequently observed. In different geographical areas, the importance of each one might vary. Outbreaks of diseases such as botulism occur occasionally and can cause important economic losses. Cattle tick fever can cause significant losses in zones of enzootic tick instability. Technical assistance and sanitary and food management are critical for the best productivity in feedlot cattle.(AU)

Esta revisão discute as principais causas de morte em bovinos de corte em confinamento. Descreve as perdas econômicas resultantes dessas mortes e sugere alternativas de controle. As doenças associadas aos sistemas respiratório e digestivo foram as mais frequentemente observadas. Em diferentes áreas geográficas, a importância de cada uma pode variar. Surtos de doenças como o botulismo ocorrem ocasionalmente e podem causar importantes perdas econômicas. A tristeza parasitária bovina pode causar perdas significativas em zonas de instabilidade enzoótica do carrapato. A assistência técnica e um bom gerenciamento sanitário e alimentar são essenciais para a melhor produtividade em bovinos de corte confinados.(AU)

Óleo essencial de Lippia gracilis: atividade antioxidante e antimicrobiana contra Clostridium botulinum / Essential oil of Lippia gracilis: antioxidant and antimicrobial activity against Clostridium botulinum

Apesar da utilização dos aditivos nos alimentos, alguns microrganismos, como o Clostridium botulinum ainda causam preocupação. Óleos essenciais são opções de aditivo para a indústria de alimentos, pois podem apresentar atividade antimicrobiana e antioxidante. Diante disso, o objetivo foi extrair o ó leo essencial de Lippia gracilis, caracterizar sua composição química por CG/ EM, avaliar a atividade antioxidante pela redução do radical DPPH e a concentração mínima esporicida (CME) sobreClostridium sporogenes. O óleo essencial é composto majoritariamente pelos monoterpenos carvacrol, p-cimeno e terpineno, e apresentou baixa capacidade de estabilização do radical DPPH, IC50 > 500 μg/ mL. Em relação à ação sobre os endósporos do Clostridium sporogenes, a concentração mínima esporicida foi 1,5% de óleo essencial.(AU)

Óleo essencial de Lippia gracilis: atividade antioxidante e antimicrobiana contra Clostridium botulinum / Essential oil of Lippia gracilis: antioxidant and antimicrobial activity against Clostridium botulinum

Apesar da utilização dos aditivos nos alimentos, alguns microrganismos, como o Clostridium botulinum ainda causam preocupação. Óleos essenciais são opções de aditivo para a indústria de alimentos, pois podem apresentar atividade antimicrobiana e antioxidante. Diante disso, o objetivo foi extrair o ó leo essencial de Lippia gracilis, caracterizar sua composição química por CG/ EM, avaliar a atividade antioxidante pela redução do radical DPPH e a concentração mínima esporicida (CME) sobreClostridium sporogenes. O óleo essencial é composto majoritariamente pelos monoterpenos carvacrol, p-cimeno e terpineno, e apresentou baixa capacidade de estabilização do radical DPPH, IC50 > 500 μg/ mL. Em relação à ação sobre os endósporos do Clostridium sporogenes, a concentração mínima esporicida foi 1,5% de óleo essencial.

Botulism in cattle associated with osteophagy in the State of Acre, Brazil

Background: Botulism is a non-febrile intoxication resulting from the ingestion of Clostridium botulinum neurotoxinsmanifested by partial or complete flaccid paralysis of the musculature of locomotion, swallowing and respiration. Theobjective of this study was to report the first case of botulinum intoxication associated with osteopathy in the state of Acre,as well as to alert breeders and veterinarians to the incidence of this disease in cattle farming.Case: The present report is an outbreak of botulism in the municipality of Acrelândia, in the state of Acre, which resultedin the death of 16 Nelore beef cattle in approximately 30 days. The affected animals were females in reproductive phasemaintained under extensive breeding system. The main clinical signs presented were weakness in the pelvic limbs, prostration, recumbency and death in less than 48 h. Only one animal, with similar symptomatology, was found alive and submitted to emergency therapeutic measures, but without success. During the necropsy of this bovine, no significant changeswere found, only related to the decubitus and agony time, except for fragments of long bones visualized in the reticulum.Samples of bone particles, ruminal contents, reticulum, rumen and intestine fragments were collected for the detection ofbotulinum toxins by the mouse bioassay method, as well as brain and brain stem for differential diagnosis of rabies andbovine spongiform encephalopathy by direct immunofluorescence and immunohistochemistry, respectively. The sampleswere sent to the Laboratory of General Bacteriology of the Biological Institute, São Paulo, Brazil, and all the analyzespresented negative results.Discussion: One of the main risk factors for the occurrence of botulinum...

Botulism in cattle associated with osteophagy in the State of Acre, Brazil

Background: Botulism is a non-febrile intoxication resulting from the ingestion of Clostridium botulinum neurotoxinsmanifested by partial or complete flaccid paralysis of the musculature of locomotion, swallowing and respiration. Theobjective of this study was to report the first case of botulinum intoxication associated with osteopathy in the state of Acre,as well as to alert breeders and veterinarians to the incidence of this disease in cattle farming.Case: The present report is an outbreak of botulism in the municipality of Acrelândia, in the state of Acre, which resultedin the death of 16 Nelore beef cattle in approximately 30 days. The affected animals were females in reproductive phasemaintained under extensive breeding system. The main clinical signs presented were weakness in the pelvic limbs, prostration, recumbency and death in less than 48 h. Only one animal, with similar symptomatology, was found alive and submitted to emergency therapeutic measures, but without success. During the necropsy of this bovine, no significant changeswere found, only related to the decubitus and agony time, except for fragments of long bones visualized in the reticulum.Samples of bone particles, ruminal contents, reticulum, rumen and intestine fragments were collected for the detection ofbotulinum toxins by the mouse bioassay method, as well as brain and brain stem for differential diagnosis of rabies andbovine spongiform encephalopathy by direct immunofluorescence and immunohistochemistry, respectively. The sampleswere sent to the Laboratory of General Bacteriology of the Biological Institute, São Paulo, Brazil, and all the analyzespresented negative results.Discussion: One of the main risk factors for the occurrence of botulinum...(AU)

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

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

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

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)

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)

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.(AU)

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.

Jürgen Döbereiner: a life dedicated to science / Jürgen Döbereiner: uma vida dedicada à ciência

Dr. Jürgen Döbereiner was born in Germany, on the 1st of November 1923, and lived in Brazil for 68 years during which time he developed a range of scientific projects in veterinary pathology and related disciplines. His main interests were the identification of new poisonous plants and mineral deficiencies and the causes of "cara inchada" ("swollen face" a periodontal disease) and botulism in livestock. This research has resulted in the improved health and saving of hundreds of thousands of animals, mainly cattle, annually, and is consequently of enormous economic value to the country. This contribution remains largely under appreciated. He was also involved in organizing diagnostic methods for identifying infectious diseases such as African swine fever and glanders in horses. One of his other major achievements has been the foundation and editing of specialized scientific journals for the documentation of veterinary science research results. At the beginning of his career in the 1950s, he and colleagues from the Institute for Animal Biology (IBA) were struggling to find a national scientific journal where research results from veterinary medicine could be published with practical application to the Brazilian reality. In consequence, the team founded "Arquivos do Instituto de Biologia Animal" and published three volumes (1959-1961). He then founded and edited "Pesquisa Agropecuária Brasileira" (The Brazilian Journal of Agricultural Research") that included a veterinary section. A series of veterinary volumes were published (1966-1976). Finally, in 1978 he helped create the Brazilian College of Veterinary Pathology (CBPA) that published "Pesquisa Veterinária Brasileira" (The Brazilian Journal of Veterinary Research) from 1981. The main goal was to communicate the most relevant disease problems of Brazilian livestock, in particular pathology and related subjects such as epidemiology, clinical study series and laboratory diagnosis to field veterinarians and academics. Dr. Jürgen Döbereiner was president of CBPA (1978-2018) and chief editor of "Pesquisa Veterinária Brasileira" (1981-2018). He passed away on the 16th of October, 2018, at the age of 94 at his home in Seropédica/RJ, Brazil.(AU)

Dr. Jürgen Döbereiner nasceu na Alemanha em 1 de novembro de 1923, durante 68 anos viveu no Brasil e desenvolveu trabalhos científicos no campo da patologia veterinária latu sensu. Sua contribuição científica de destaque foi em temas como plantas tóxicas de interesse pecuário, deficiências minerais em animais de produção, cara inchada (doença periodontal) dos ruminantes, botulismo e diagnóstico de doenças infecciosas. Estas pesquisas resultaram na melhoria da saúde e de centenas de milhares de animais, principalmente bovinos e, consequentemente, foram de enorme valor econômico para o país. Esta contribuição ainda permanece em grande parte subestimada. De grande destaque para a ciência brasileira foi ainda a sua atuação profissional na documentação científica de resultados de pesquisa. No início de sua carreira na década de 1950, Dr. Döbereiner e outros pesquisadores do Instituto de Biologia Animal (IBA) detectaram a necessidade de um periódico científico nacional para publicar resultados de pesquisas com aplicação pratica à realidade brasileira. Dessa iniciativa surgiram os Arquivos do Instituto de Biologia Animal, que publicou três fascículos (1959-1961), em seguida o Dr. Jürgen Döbereiner participou na fundação da revista Pesquisa Agropecuária Brasileira que publicou a Série Veterinária (1966-1976) e finalmente em 1978, houve a fundação do Colégio Brasileiro de Patologia Animal (CBPA) que publica desde 1981 a revista Pesquisa Veterinária Brasileira. Este periódico científico foi criado para apresentar à comunidade, principalmente veterinários de campo e professores, os principais problemas de saúde em animais de produção no Brasil, ou seja, patologia em seu sentido amplo, envolvendo as áreas de epidemiologia, clínica e diagnóstico laboratorial. Dr. Jürgen Döbereiner, que foi presidente do CBPA (1978-2018) e Editor-Chefe da revista Pesquisa Veterinária Brasileira (1981-2018), faleceu em casa, em 16 de outubro de 2018, aos 94 anos, no município de Seropédica/RJ.(AU)