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)

Dietary supplementation of Agaricus bisporus stalk meal on growth performance, carcass and organ traits, meat quality, cecum mesophilic aerobic bacteria counts and intestinal histology in broiler chickens / Efeitos da suplementação dietética de farelo de caule de Agaricus bisporus no desempenho de crescimento, características de carcaça e órgãos, qualidade da carne, contagem de bactérias aeróbias mesófilas cecais e histologia intestinal em frangos de corte

Effects of dietary Agaricus bisporus mixture or stalk or cap on growth performance, carcass components and some meat quality parameters, mesophilic aerobic bacterial counts, and intestinal histomorphology in broiler chickens were investigated. Two hundred and forty one-day-old male Ross 308 broiler chickens were divided into 4 experimental groups with 4 replicates, each including 15 birds. Chickens were fed with basal diet (C), mushroom mixture (MM, 10 g stalk+10 g cap/kg diet), mushroom stalk (MS, 20 g/kg diet) and mushroom cap (MC, 20 g/kg diet). Feed conversion ratio (FCR) was improved (P < 0.01) by dietary MS while feed intake (FI) decreased (P < 0.01) in all treatment groups compared to control. However, body weight gain (BWG) was decreased (P < 0.01) by MC inclusion. Mushroom supplemented groups had higher (P < 0.05) mesophilic aerobic bacteria in the cecum. MS inclusion increased villus height to crypt depth ratio in the jejunum (P < 0.01) and villus height in the ileum (P < 0.01) and jejunum (P < 0.05). MC increased (P < 0.01) crypt depth in the jejunum. There were no statistical differences among groups for carcass components (P > 0.05). The L* values were decreased (P < 0.01) in thigh meat but increased (P < 0.01) in breast meat by mushroom inclusion. The a* values were decreased (P < 0.01) and b* values were increased (P < 0.05) in both thigh and breast meat with mushroom addition to diet. The obtained results indicated that A. bisporus stalk meal at an inclusion level of 20 g/kg of diet had favourable effects on growth performance with higher feed efficiency, improved intestinal morphology with higher villus height and increased meat quality of broiler chickens.

Foram investigados os efeitos da mistura dietética de Agaricus bisporus ou talo ou tampa sobre o desempenho de crescimento, componentes da carcaça e alguns parâmetros de qualidade da carne, contagens de bactérias aeróbias mesofílicas e histomorfologia intestinal em frangos de corte. Duzentos e quarenta frangos de corte Ross 308 de um dia de idade foram divididos em quatro grupos experimentais com quatro repetições, cada um incluindo 15 aves. Galinhas alimentadas com dieta basal (C), mistura de cogumelos (MM, 10 g caule + 10 g cap / kg de dieta), caule de cogumelo (MS, 20 g / kg dieta) e cogumelo (MC, 20 g / kg dieta). A taxa de conversão alimentar (FCR) foi melhorada (P < 0.01) por MS dietético enquanto o consumo de ração (FI) diminuiu (P < 0.01) em todos os grupos de tratamento em comparação com o controle. No entanto, o ganho de peso corporal (BWG) foi diminuído (P < 0.01) pela inclusão de MC. Os grupos suplementados com cogumelos apresentaram maiores (P < 0.05) bactérias aeróbias mesofílicas no ceco. A inclusão de MS aumentou a proporção entre a largura das vilosidades e a profundidade da cripta no jejuno (P < 0.01) e o comprimento das vilosidades no íleo (P < 0.01) e jejuno (P < 0.05). MC aumentou (P < 0.01) a profundidade de cripta no jejuno. Não houve diferença estatística entre os grupos nos componentes da carcaça (P > 0.05). Os valores L * diminuíram (P < 0.01) na carne da coxa, mas aumentaram (P < 0.01) na carne de peito com a inclusão de cogumelos. Os valores de a* diminuíram (P < 0.01) e os valores de b* aumentaram (P < 0.05) em ambas as coxas e peitos pela adição de cogumelos à dieta. Os resultados obtidos indicaram que a farinha de colmo de A. bisporus com um nível de inclusão de 20 g / kg de dieta teve efeitos favoráveis no desempenho do crescimento com maior eficiência alimentar, melhorou a morfologia intestinal com maior comprimento de vilosidade e aumentou a qualidade da carne de frangos de corte.

Direct Detection and Quantification of Bacterial Pathogens from Broiler Cecal Samples in the Slaughter Line by Real-Time PCR

Chicken meat is an important source of foodborne pathogens, including Salmonella, Campylobacter, and Clostridium perfringens. These bacteria can occur in the intestinal microbiota of broilers and contaminate chicken carcasses in industrial meat processing. This study aimed to develop and evaluate a procedure based on real-time PCRs for the direct detection and quantification of these three bacteria in broilers' ceca collected in poultry slaughter houses and demonstrate the occurrence of these important foodborne pathogens in Brazilian poultry production flocks. Cecal contents were collected from 45 different broiler flocks in three different slaughterhouses in the state of Paraná, Brazil, totaling 45 samples (in pools of 10 different ceca/chickens per broiler flock). Then, these samples were tested for the detection and quantification of Salmonella, Campylobacter, and Clostridium perfringens by real-time PCRs. The results demonstrated the occurrence of three (6.7%) positive pools for Salmonella, 20 (44.4%) for Campylobacter, and 32 (71.1%) for C. perfringens. Mean bacterial concentrations in the positive samples were 4.3log10 cells/g for Salmonella, 6.4 log10 cells/g for Campylobacter, and 5.5 log10 cells/g for C. perfringens. In conclusion, Salmonella, Campylobacter, and C. perfringens could be detected and quantified directly from the broilers cecal contents collected in the slaughter line. This procedure will be certainly useful to more quickly detect these foodborne pathogens and prevent their occurrence in chicken meat and other poultry food products.(AU)

The effect of hydrated sodium calcium aluminosilicate on fatty liver and the composition of the intestinal microbiota in overfed landes geese

Goose fatty liver is a delicious food product and the overfeeding will cause the abnormal physiology of the geese. The objective of this study was to investigate the effect of supplementation with hydrated sodium calcium aluminosilicate (HSCAS) on the fatty liver, ileal and cecal microbiota of Landes geese during overfeeding. Sixty 70-day-old Landes geese (body weight= 3.0 ± 0.05 kg) were randomly divided into three groups, two of which were overfed with whole corn supplemented with or without HSCAS for 20 days when the fatty liver reaches to the maximum size and the negative control group was ad libitum access to the corn basal diet. The intestinal contents of the ileum and cecum from three geese per group were used for high-throughput sequencing. As a result of this study, the HSCAS-treatment led to an increase in relative liver weight (p 0.05) of geese compared with the overfeeding control group. The richness and diversity of the bacterial communities decreased in the ileum and ceca after overfeeding. Overfeeding increased the relative abundance of Firmicutes, especially Lactobacillus, in ileal samples. HSCAS supplementation increased the relative abundance of Lactobacillus, and decreased the relative abundance of Actinobacillus in the ileum and the relative abundance of Erysipelotrichi, Bacteroides and Escherichia in the ceca. Bacterial richness indicators were also increased in samples from ileum and ceca after HSCAS supplementation. In conclusion, dietary HSCAS supplementation promoted liver performance in overfed Landes geese. HSCAS treatment had a beneficial effect on the intestinal microbiota composition in geese during the overfeeding.(AU)

The effect of hydrated sodium calcium aluminosilicate on fatty liver and the composition of the intestinal microbiota in overfed landes geese

Goose fatty liver is a delicious food product and the overfeeding will cause the abnormal physiology of the geese. The objective of this study was to investigate the effect of supplementation with hydrated sodium calcium aluminosilicate (HSCAS) on the fatty liver, ileal and cecal microbiota of Landes geese during overfeeding. Sixty 70-day-old Landes geese (body weight= 3.0 ± 0.05 kg) were randomly divided into three groups, two of which were overfed with whole corn supplemented with or without HSCAS for 20 days when the fatty liver reaches to the maximum size and the negative control group was ad libitum access to the corn basal diet. The intestinal contents of the ileum and cecum from three geese per group were used for high-throughput sequencing. As a result of this study, the HSCAS-treatment led to an increase in relative liver weight (p 0.05) of geese compared with the overfeeding control group. The richness and diversity of the bacterial communities decreased in the ileum and ceca after overfeeding. Overfeeding increased the relative abundance of Firmicutes, especially Lactobacillus, in ileal samples. HSCAS supplementation increased the relative abundance of Lactobacillus, and decreased the relative abundance of Actinobacillus in the ileum and the relative abundance of Erysipelotrichi, Bacteroides and Escherichia in the ceca. Bacterial richness indicators were also increased in samples from ileum and ceca after HSCAS supplementation. In conclusion, dietary HSCAS supplementation promoted liver performance in overfed Landes geese. HSCAS treatment had a beneficial effect on the intestinal microbiota composition in geese during the overfeeding.

Glicerina associada à ureia na terminação de bovinos: parâmetros ruminais, digestibilidade e massa microbiana / Glycerin associated with urea in finishing cattle: ruminal fermentation, digestibility and microbial mass

O objetivo desta pesquisa foi verificar a combinação de glicerina com ureia, quanto aos parâmetros ruminais, à digestibilidade e à qualidade da massa microbiana. O trabalho foi conduzido na Unidade Animal de Estudos Digestivos e Metabólicos, pertencente à Faculdade de Ciências Agrárias e Veterinárias-FCAV/Unesp, Câmpus de Jaboticabal. Seis animais da raça Nelore providos de cânulas ruminais foram utilizados para verificar os parâmetros pH, nitrogênio amoniacal, a digestibilidade e a massa microbiana, distribuídos em quadrado latino 6x6. Seis dietas com energia metabolizável e proteína bruta semelhantes foram formuladas, utilizando-se a proporção volumoso:concentrado de 30:70. O volumoso utilizado foi silagem de milho. Os concentrados utilizados foram compostos por milho, casca de soja, farelo de girassol, glicerina e ureia. Os tratamentos foram: C = controle; U = 1% de ureia; G10 = 10% de glicerina; GU10 = 1% de ureia e 10% de glicerina, G20 = 20% de glicerina e GU20 = 20% de glicerina e 1% de ureia. Houve influência das dietas no crescimento dos microrganismos ruminais, no pH e no nitrogênio amoniacal. Dietas com 20% de glicerina na matéria seca da dieta diminuem a população de protozoários ruminais. A inclusão de glicerina diminui a digestibilidade de componentes fibrosos da dieta.(AU)

The aim of this research was to investigate the combination of glycerol with urea as the ruminal parameters, digestibility and quality of microbial mass. The work was conducted at the Animal Unit of Digestive and Metabolic Studies belonging to the Faculty of Agricultural Sciences and Veterinárias-FCAV/Unesp campus Jaboticabal. Six animals fitted with rumen cannulas Nellore were used to verify the parameters pH, ammonia nitrogen, digestibility, and microbial mass distributed in a 6x6 Latin square. Six diets with similar metabolizable energy and crude protein were formulated using the forage:concentrate ratio of 30:70. The roughage used was corn silage. The concentrates used were composed of corn, soybean hulls, sunflower meal, glycerin and urea. The treatments were: C = control, U = 1 % urea, G10 = 10 % glycerin, GU10 = 1 % urea and 10 % glycerol, G20 = 20 % glycerin and GU20 = 20 % glycerin and 1 % urea. Diets with glycerin had lower digestibility of NDF and hemicellulose. There was no influence of diets on the growth of rumen microorganisms, pH and ammonia nitrogen. Diets with 20 % glycerin in the diet dry matter decrease the population of rumen protozoa. The addition of glycerol decreases the digestibility of fibrous components of the diet.(AU)

Glicerina associada à ureia na terminação de bovinos: parâmetros ruminais, digestibilidade e massa microbiana / Glycerin associated with urea in finishing cattle: ruminal fermentation, digestibility and microbial mass

O objetivo desta pesquisa foi verificar a combinação de glicerina com ureia, quanto aos parâmetros ruminais, à digestibilidade e à qualidade da massa microbiana. O trabalho foi conduzido na Unidade Animal de Estudos Digestivos e Metabólicos, pertencente à Faculdade de Ciências Agrárias e Veterinárias-FCAV/Unesp, Câmpus de Jaboticabal. Seis animais da raça Nelore providos de cânulas ruminais foram utilizados para verificar os parâmetros pH, nitrogênio amoniacal, a digestibilidade e a massa microbiana, distribuídos em quadrado latino 6x6. Seis dietas com energia metabolizável e proteína bruta semelhantes foram formuladas, utilizando-se a proporção volumoso:concentrado de 30:70. O volumoso utilizado foi silagem de milho. Os concentrados utilizados foram compostos por milho, casca de soja, farelo de girassol, glicerina e ureia. Os tratamentos foram: C = controle; U = 1% de ureia; G10 = 10% de glicerina; GU10 = 1% de ureia e 10% de glicerina, G20 = 20% de glicerina e GU20 = 20% de glicerina e 1% de ureia. Houve influência das dietas no crescimento dos microrganismos ruminais, no pH e no nitrogênio amoniacal. Dietas com 20% de glicerina na matéria seca da dieta diminuem a população de protozoários ruminais. A inclusão de glicerina diminui a digestibilidade de componentes fibrosos da dieta.(AU)

The aim of this research was to investigate the combination of glycerol with urea as the ruminal parameters, digestibility and quality of microbial mass. The work was conducted at the Animal Unit of Digestive and Metabolic Studies belonging to the Faculty of Agricultural Sciences and Veterinárias-FCAV/Unesp campus Jaboticabal. Six animals fitted with rumen cannulas Nellore were used to verify the parameters pH, ammonia nitrogen, digestibility, and microbial mass distributed in a 6x6 Latin square. Six diets with similar metabolizable energy and crude protein were formulated using the forage:concentrate ratio of 30:70. The roughage used was corn silage. The concentrates used were composed of corn, soybean hulls, sunflower meal, glycerin and urea. The treatments were: C = control, U = 1 % urea, G10 = 10 % glycerin, GU10 = 1 % urea and 10 % glycerol, G20 = 20 % glycerin and GU20 = 20 % glycerin and 1 % urea. Diets with glycerin had lower digestibility of NDF and hemicellulose. There was no influence of diets on the growth of rumen microorganisms, pH and ammonia nitrogen. Diets with 20 % glycerin in the diet dry matter decrease the population of rumen protozoa. The addition of glycerol decreases the digestibility of fibrous components of the diet.(AU)

Prebiótico, probiótico e simbiótico para larvas de Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae) / Prebiotic, probiotic and symbiotic for Trichogaster leeri larvae (Bleeker, 1852, Perciformes, Osphronemidae)

Avaliou-se a suplementação de prebiótico (mananoligossacarídeo - 2g/kg), probiótico (Bacillus subtilis, Cohn, 1872, Bacillales, Bacillaceae - 2g/kg) e simbiótico (mananoligossacarídeo - 2g/kg + B. subtilis - 2g/kg), em rações para larvas de Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae), durante a transição alimentar, sobre o crescimento, a sobrevivência, a morfometria intestinal e a resistência ao estresse por exposição aérea. Para isso, 420 larvas (0,45±0,05mg) foram distribuídas em 20 aquários (3,5L), em delineamento inteiramente ao acaso, com cinco tratamentos e quatro repetições, durante 28 dias, a saber: 12 dias de alimento vivo (AV) + oito dias de coalimentação (AV + ração controle) + oito dias de ração controle; 12 dias de AV + oito dias de coalimentação (AV + ração prebiótico) + oito dias de ração prebiótico; 12 dias de AV + oito dias de coalimentação (AV + ração probiótico) + oito dias de ração probiótico; 12 dias de AV + oito dias de coalimentação (AV + ração simbiótico) + oito dias de ração simbiótico; 28 dias de AV. Larvas que receberam apenas AV apresentaram resultados de desempenho zootécnico significativamente superiores às larvas dos demais tratamentos, exceto para taxa de crescimento específico em comprimento e sobrevivência, que foram semelhantes aos resultados obtidos por larvas que receberam rações suplementadas com simbiótico. Larvas que receberam a ração controle apresentaram resultados significativamente inferiores para altura das vilosidades intestinais e taxa de resistência ao estresse comparando-se aos demais tratamentos. Os resultados deste estudo indicam que a suplementação com prebiótico, probiótico e simbiótico exerce efeito positivo sobre a sobrevivência, a morfometria intestinal e a resistência ao estresse em larvas de T. leeri, no entanto somente a suplementação com simbiótico resulta em melhora no crescimento.(AU)

The prebiotic (mannan oligosaccharide - 2g/kg), probiotic (Bacillus subtilis Cohn, 1872, Bacillales, Bacillaceae - 2g/kg) and symbiotic (mannan oligosaccharide - 2g/kg + Bacillus subtilis - 2g/kg) supplementation in diets during food transition in Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae) larvae on growth, survival, intestinal morphology and stress resistance to air exposure was evaluated. For this, 420 larvae (0.45±0.05mg) were distributed in 20 aquaria (3.5L) in a completely randomized design with five treatments and four replications for 28 days, namely: 12 days of live food (LF) + eight days of co-feeding (LF + control diet) + eight days of control diet; 12 days of LF + eight days of co-feeding (LF + prebiotic feed) + eight days of prebiotic feed; 12 days of LF + eight days of co-feeding (LF + probiotic feed) + eight days of probiotic feed; 12 days of LF + eight days of co-feeding (LF + symbiotic feed) + eight days symbiotic feed; 28 days of LF. Larvae fed only LF showed growth performance significantly higher than the larvae of other treatments, except for specific growth rate in length and survival that were similar to the results obtained by larvae fed diets symbiotically supplemented. Larvae fed the control diet showed significantly lower results for height of intestinal villi and stress resistance rate comparing to other treatments. The results of this study indicate that prebiotic, probiotic and symbiotic supplementation has a positive effect on survival, intestinal morphology and resistance to stress in larvae of T. leeri, however, only symbiotic supplementation results in improved growth.(AU)

Prebiótico, probiótico e simbiótico para larvas de Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae) / Prebiotic, probiotic and symbiotic for Trichogaster leeri larvae (Bleeker, 1852, Perciformes, Osphronemidae)

Avaliou-se a suplementação de prebiótico (mananoligossacarídeo - 2g/kg), probiótico (Bacillus subtilis, Cohn, 1872, Bacillales, Bacillaceae - 2g/kg) e simbiótico (mananoligossacarídeo - 2g/kg + B. subtilis - 2g/kg), em rações para larvas de Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae), durante a transição alimentar, sobre o crescimento, a sobrevivência, a morfometria intestinal e a resistência ao estresse por exposição aérea. Para isso, 420 larvas (0,45±0,05mg) foram distribuídas em 20 aquários (3,5L), em delineamento inteiramente ao acaso, com cinco tratamentos e quatro repetições, durante 28 dias, a saber: 12 dias de alimento vivo (AV) + oito dias de coalimentação (AV + ração controle) + oito dias de ração controle; 12 dias de AV + oito dias de coalimentação (AV + ração prebiótico) + oito dias de ração prebiótico; 12 dias de AV + oito dias de coalimentação (AV + ração probiótico) + oito dias de ração probiótico; 12 dias de AV + oito dias de coalimentação (AV + ração simbiótico) + oito dias de ração simbiótico; 28 dias de AV. Larvas que receberam apenas AV apresentaram resultados de desempenho zootécnico significativamente superiores às larvas dos demais tratamentos, exceto para taxa de crescimento específico em comprimento e sobrevivência, que foram semelhantes aos resultados obtidos por larvas que receberam rações suplementadas com simbiótico. Larvas que receberam a ração controle apresentaram resultados significativamente inferiores para altura das vilosidades intestinais e taxa de resistência ao estresse comparando-se aos demais tratamentos. Os resultados deste estudo indicam que a suplementação com prebiótico, probiótico e simbiótico exerce efeito positivo sobre a sobrevivência, a morfometria intestinal e a resistência ao estresse em larvas de T. leeri, no entanto somente a suplementação com simbiótico resulta em melhora no crescimento.(AU)

The prebiotic (mannan oligosaccharide - 2g/kg), probiotic (Bacillus subtilis Cohn, 1872, Bacillales, Bacillaceae - 2g/kg) and symbiotic (mannan oligosaccharide - 2g/kg + Bacillus subtilis - 2g/kg) supplementation in diets during food transition in Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae) larvae on growth, survival, intestinal morphology and stress resistance to air exposure was evaluated. For this, 420 larvae (0.45±0.05mg) were distributed in 20 aquaria (3.5L) in a completely randomized design with five treatments and four replications for 28 days, namely: 12 days of live food (LF) + eight days of co-feeding (LF + control diet) + eight days of control diet; 12 days of LF + eight days of co-feeding (LF + prebiotic feed) + eight days of prebiotic feed; 12 days of LF + eight days of co-feeding (LF + probiotic feed) + eight days of probiotic feed; 12 days of LF + eight days of co-feeding (LF + symbiotic feed) + eight days symbiotic feed; 28 days of LF. Larvae fed only LF showed growth performance significantly higher than the larvae of other treatments, except for specific growth rate in length and survival that were similar to the results obtained by larvae fed diets symbiotically supplemented. Larvae fed the control diet showed significantly lower results for height of intestinal villi and stress resistance rate comparing to other treatments. The results of this study indicate that prebiotic, probiotic and symbiotic supplementation has a positive effect on survival, intestinal morphology and resistance to stress in larvae of T. leeri, however, only symbiotic supplementation results in improved growth.(AU)

Performance, intestinal morphology and microbiology of broiler chickens fed egg powder in the starter diet

The effect of egg powder inclusion in the pre-starter diet (1-7 days of age) on the performance, and intestinal morphology and microbiology of male broiler chickens was evaluated in a completely randomized design. Starter diets with equal metabolizable energy and crude protein levels were formulated to contain 0, 20, 40, or 60 g egg powder/kg diet. Results showed that body weight, feed intake, feed conversion ratio, and European efficiency index were positively affected (p0.05) by egg powder inclusion in the starter diet. Inclusion of 40 g egg powder/kg diet in the starter diet promoted the best performance results. Jejunal villus height and villus to crypt ratio increased (p0.05) and intestinal length decreased (p0.05) as a result of egg powder inclusion in the starter diet. Egg powder inclusion in the starter diet reduced (p0.05E. coli counts, but increased (p0.05) lactic acid bacteria counts in the small intestine. The results obtained in the present study indicate that the inclusion of 40 60 g egg powder/kg of starter diet g improved the performance and intestinal health of broilers.

Performance, intestinal morphology and microbiology of broiler chickens fed egg powder in the starter diet

The effect of egg powder inclusion in the pre-starter diet (1-7 days of age) on the performance, and intestinal morphology and microbiology of male broiler chickens was evaluated in a completely randomized design. Starter diets with equal metabolizable energy and crude protein levels were formulated to contain 0, 20, 40, or 60 g egg powder/kg diet. Results showed that body weight, feed intake, feed conversion ratio, and European efficiency index were positively affected (p0.05) by egg powder inclusion in the starter diet. Inclusion of 40 g egg powder/kg diet in the starter diet promoted the best performance results. Jejunal villus height and villus to crypt ratio increased (p0.05) and intestinal length decreased (p0.05) as a result of egg powder inclusion in the starter diet. Egg powder inclusion in the starter diet reduced (p0.05E. coli counts, but increased (p0.05) lactic acid bacteria counts in the small intestine. The results obtained in the present study indicate that the inclusion of 40 60 g egg powder/kg of starter diet g improved the performance and intestinal health of broilers. (AU)

Detection of carboxylesterase and esterase activity in culturable gut bacterial flora isolated from diamondback moth, Plutella xylostella (Linnaeus), from India and its possible role in indoxacarb degradation

Diamondback moth (DBM), Plutella xylostella (Linnaeus), is a notorious pest of brassica crops worldwide and is resistant to all groups of insecticides. The insect system harbors diverse groups of microbiota, which in turn helps in enzymatic degradation of xenobiotic-like insecticides. The present study aimed to determine the diversity of gut microflora in DBM, quantify esterase activity and elucidate their possible role in degradation of indoxacarb. We screened 11 geographic populations of DBM in India and analyzed them for bacterial diversity. The culturable gut bacterial flora underwent molecular characterization with 16S rRNA. We obtained 25 bacterial isolates from larvae (n = 13) and adults (n = 12) of DBM. In larval gut isolates, gammaproteobacteria was the most abundant (76%), followed by bacilli (15.4%). Molecular characterization placed adult gut bacterial strains into three major classes based on abundance: gammaproteobacteria (66%), bacilli (16.7%) and flavobacteria (16.7%). Esterase activity from 19 gut bacterial isolates ranged from 0.072 to 2.32 µmol/min/mg protein. Esterase bands were observed in 15 bacterial strains and the banding pattern differed in Bacillus cereus KC985225 and Pantoea agglomerans KC985229. The bands were characterized as carboxylesterase with profenofos used as an inhibitor. Minimal media study showed that B. cereus degraded indoxacarb up to 20%, so it could use indoxacarb for metabolism and growth. Furthermore, esterase activity was greater with minimal media than control media: 1.87 versus 0.26 µmol/min/mg protein. Apart from the insect esterases, bacterial carboxylesterase may aid in the degradation of insecticides in DBM.(AU)

Análise da microbiota gastrointestinal de Chelonoidis provenientes de cativeiro da região noroeste de São Paulo / Microbiota analysis of gastrointestinal Chelonoidis from captivity in the northwestern region of São Paulo / Análisis de la microflora gastrointestinal de Chelonoidis coming from cautividad de región noroeste de São Paulo

O jabuti é a designação vulgar dos répteis do género Chelonoidis, da ordem dos Quelônios, da família dos testudinídeos. A literatura apresenta escassas informações a respeito da importância desse réptil silvestre na cadeia epidemiológica das infecções. A determinação da microbiota entérica de animais silvestres tem por importância não só o conhecimento de micro-organismos que venham a compor a flora intestinal normal dessas espécies, como também a possibilidade de se descobrir novos agentes veiculadores de micro-organismos patogênicos ao homem, e para as demais espécies animais. Com a finalidade de verificar a microbiota de Chelonoidis, foram colhidos "swabs" retais de nove membros desta espécie que se encontram confinados no Zoológico Municipal da cidade de Araçatuba. O material foi enviado e processado no laboratório de Microbiologia da Faculdade de Medicina Veterinária UNESP - Campus de Araçatuba. As amostras foram semeadas em meios de Ágar Sangue e MacConkey incubadas em atmosfera de aerobiose à temperatura de 37°C por 24h, e em ágar Sabouraud que permaneceram em anaerobiose por 7 a 15 dias. Características morfológicas e tintoriais fundamentaram a identificação das colônias bacterianas, além das provas bioquímicas. Das amostras analisadas somente 55,5% obtiveram resultados conclusivos, sendo identificadas prevalências de 100% e 20% para Escherichia coli e a Klebsiella oxytoca, respectivamente. Os resultados revelam que geoquelônios atuam significativamente na ecoepidemiologia de doenças infecciosas com a manutenção e a propagação de patógenos no ambiente.

The jabuti is the common name of the genus Chelonoidis reptiles of the order of turtles, the family of testudinídeos. The literature presents little information on the importance of reptile in the wild epidemiological chain of infection. The determination of the enteric microbiota of wild animals is important not only to knowledge of microorganisms that will form the normal intestinal flora of these species, but also the possibility to discover new agents of pathogenic microorganisms to humans and to other animal species. In order to verify the microbiota of Chelonoidis, were collected "swabs" rectal nine members of this species that are confined in the Municipal Zoo of the city of Araçatuba. The material was sent and processed in the laboratory of Microbiology, Faculty of Veterinary Medicine UNESP - Campus de Araçatuba. The samples were sown in media of blood and MacConkey agar incubated in aerobic atmosphere at a temperature of 37 °C for 24h, and Sabouraud agar remained in anaerobiosis for 7 to 15 days. Morphological characteristics and staining based identification of bacterial colonies, in addition to the biochemical evidence. Samples of only 55.5% obtained conclusive results, and identified prevalence of 100% and 20% for Escherichia coli and Klebsiella oxytoca, respectively. The results show that significantly geoquelônios work in Eco-epidemiology of infectious diseases with the maintenance and spread of pathogens in the environment.

El jabuti es una especie de tortuga del género Chelonoidis, de la familia de los testudinídeos. En la literatura hay poca información acerca de la naturaleza de estos reptiles en la cadena epidemiológica de la infección. La determinación de la microflora entérica de los animales silvestres es importante no sólo para el conocimiento de los microorganismos que forman la flora intestinal normal de estas especies, sino también en la posibilidad de descubrir nuevos agentes de microorganismos patógenos para los seres humanos y otras especies animales. Con la finalidad de verificar la microflora de Chelonoidis se tomarán "frotis" del recto de esta especie de tortugas las cuales se encuentran en el zoológico municipal. El material fue enviado y procesado en el Laboratorio de Microbiología, Facultad de Medicina Veterinaria de la UNESP - Campus de Araçatuba. Las muestras fueron manejadas en medios de cultivo para bacterias (agar sangre, Mac Conkey y Sabouraud) se incubaron en una ambiente para bacterias aerobias y anaerobias a una temperatura de 37°C. Serrealizaron pruebas bioquímicas para identificación bacteriana. Las muestras fueron sembradas en medios enriquecidos de agar sangre y MacConkey incubadas en atmósfera aeróbica a una temperatura de 37 ° C durante 24 horas, para luego ser mantenidas en agar Sabouraud en anaerobiosis por 7 a 15 días. La identificación de colonias bacterianas fue realizada en base a sus características morfológicas, de tinción y a través de las pruebas bioquímicas. De las muestras analizadas, sólo en el 55,5% se obtuvieron resultados concluyentes y se logro determinar la prevalencia de Escherichiacoli y Klebsiella oxytoca en un 100% y 20% respectivamente, de los casos analizados. Los resultados obtenidos en este trabajo muestran que los geoquelônios "influyen" significativamente en Eco-epidemiología de las enfermedades infecciosas con la propagación y el mantenimiento de los agentes patógenos en el medio ambiente.

Ácido fumárico em dietas para juvenis de tilápia-do-nilo

O objetivo do presente estudo foi avaliar o uso de ácido fumárico como aditivo alimentar para dietas de alevinos de tilápia-do-nilo. Seis dietas isocalóricas (3,4 kcal de energia bruta / g) e isonitrogenadas (29% de proteína bruta) suplementadas com ácido fumárico (0,0, 0,5, 1,0, 1,5, 2,0 e 3,0% da dieta) foram formuladas com quatro repetições cada e os animais foram distribuídos em delineamento inteiramente casualizado. Treze peixes (± 1,7 g) foram distribuídos em tanques e alimentados às 8:00, 12:00 e 16:00 horas até a saciedade aparente durante 35 dias. Todos os dados de desempenho zootécnico e contagem de bactérias viáveis totais (UFC-1) foram submetidos à análise de variância e as diferenças entre as médias foram avaliadas pelo teste de Tukey (p <0,05). Redução significativa de bactérias gram-negativas e comunidades de Enterobacteriaceae foi observada em peixes alimentados com dieta suplementada com ácido fumárico a 1,5% (p <0,05). Da mesma forma, ganho de peso, eficiência alimentar, taxa de crescimento específico e eficiência protéica foram numericamente maiores no tratamento de 1,5% comparado ao grupo controle e outros tratamentos (p <0,05). Por outro lado, não houve diferença significativa no consumo diário de ração (p> 0,05). A suplementação de 1,5% de ácido fumárico melhora o desempenho zootécnico e resulta em efeito antibacteriano em tilápias do Nilo.

The objective of the present study was to evaluate the use of fumaric acid as a feed additive for Nile tilapia fry diets. Six iso-caloric (3.4 kcal gross energy/ g) and iso-nitrogenous (29% crude protein) diets supplemented with fumaric acid (0.0, 0.5, 1.0, 1.5, 2.0 and 3.0% of diet) were formulated with four replicate each and the animals were distributed in a completely randomized design. Thirteen fish (± 1.7g) were distributed in tanks and fed at 8:00 a.m., 12:00 a.m. and 4:00 p.m. up to apparent satiety during 35 days. All zootechnical performance and total viable bacterial count (CFUg-1) data were subjected to analysis of variance and differences between means were assessed by Tukey's test (p<0.05). Significant reduction of gram-negative bacteria and Enterobacteriaceae communities were observed in fish fed diet supplemented with fumaric acid at 1.5% (p<0.05). Similarly, weight gain, feed efficiency, specific growth rate and protein efficiency was numerically higher in 1.5% treatment compared with control group and other treatments (p<0.05). On the other hand, there was no significant difference in daily feed intake (p>0.05). The supplementation of 1.5% fumaric acid improves zootechnical performance and results an antibacterial effect in Nile tilapia fry.