HAL (Le Centre pour la Communication Scientifique Directe), Oct 13, 2021
International audienceEnterotoxigenic Escherichia coli (ETEC) is the principal pathogen responsib... more International audienceEnterotoxigenic Escherichia coli (ETEC) is the principal pathogen responsible for post-weaning diarrhea in newly weaned piglets. Expansion of ETEC at weaning is thought to be the consequence of various stress factors such as transient anorexia, dietary change or increase in intestinal inflammation and permeability, but the exact mechanisms remain to be elucidated. As the use of animal experiments raise more and more ethical concerns, we used a recently developed in vitro model of piglet colonic microbiome and mucobiome, the MPigut-IVM, to evaluate the effects of a simulated weaning transition and pathogen challenge at weaning. Our data suggested that the tested factors impacted the composition and functionality of the MPigut-IVM microbiota. The simulation of weaning transition led to an increase in relative abundance of the Prevotellaceae family, which was further promoted by the presence of the ETEC strain. In contrast, several beneficial families such as Bacteroidiaceae or Ruminococcaceae and gut health related short chain fatty acids like butyrate or acetate were reduced upon simulated weaning. Moreover, the incubation of MPigut-IVM filtrated effluents with porcine intestinal cell cultures showed that ETEC challenge in the in vitro model led to an increased expression of pro-inflammatory genes by the porcine cells. This study provides insights about the etiology of a dysbiotic microbiota in post-weaning piglets and showed that in vitro MPigut-IVM can be efficiently coupled to porcine epithelial cells to study host-microbiota crosstalk.P075 Periparturient Holstein cows of varying subacute ruminal acidosis phenotype differ in rumen bacterial community an
bioRxiv (Cold Spring Harbor Laboratory), Oct 15, 2021
Colostrum quality is of paramount importance in the management of optimal ruminant growth and inf... more Colostrum quality is of paramount importance in the management of optimal ruminant growth and infectious disease prevention in early life. Live yeast supplementation effect during the last month of gestation was evaluated on ewes' colostrum composition. Two groups of ewes (n=14) carrying twin lambs were constituted and twins were separated into groups (mothered or artificially-fed) 12h after birth. Nutrient, oligosaccharides (OS), IgG and lactoferrin concentrations were measured over 72h after lambing, and bacterial community was described in colostrum collected at parturition (T0). Immune passive transfer was evaluated through IgG measurement in lamb serum. In both groups, colostral nutrient, OS concentrations and IgG concentrations in colostrum and lamb serum decreased over time, (p < 0.01) except for lactose, which slightly increased (p < 0.001) and lactoferrin which remained stable. Bacterial population was stable over time with high relative abundances of Aerococcaceae, Corynebacteriaceae, Moraxellaceae and Staphylococcaceae in T0-colostrum. No effect of supplementation was observed in nutrient and lactoferrin concentrations. In supplemented ewes, colostral IgG level was higher at T0 and a higher level of serum IgG was observed in lambs born from supplemented mothers and artificially-fed, while no effect of supplementation was observed in the mothered lambs groups. Using a metabolomic approach, we showed that supplementation affected OS composition with significantly higher levels of colostral Neu-5Gc compounds up to 5h after birth. No effect of supplementation was observed on bacterial composition. Our data suggest that live yeast supplementation offsets the negative impact of early separation and incomplete colostrum feeding in neonate lambs.
Colostrum quality is of paramount importance in the management of optimal ruminant growth and inf... more Colostrum quality is of paramount importance in the management of optimal ruminant growth and infectious disease prevention in early life. Live yeast supplementation effect during the last month of gestation was evaluated on ewes' colostrum composition. Two groups of ewes (n = 14) carrying twin lambs were constituted and twins were separated into groups (mothered or artificially fed) 12 h after birth. Nutrient, oligosaccharides (OS), IgG and lactoferrin concentrations were measured over 72 h after lambing, and bacterial community was described in colostrum collected at parturition (T0). Immune passive transfer was evaluated through IgG measurement in lamb serum. In both groups, colostral nutrient, OS concentrations and IgG concentrations in colostrum and lamb serum decreased over time (P < 0⋅01), except for lactose, which slightly increased (P < 0⋅001), and lactoferrin, which remained stable. Bacterial population was stable over time with high relative abundances of Aerococcaceae, Corynebacteriaceae, Moraxellaceae and Staphylococcaceae in T0 colostrum. No effect of supplementation was observed in nutrient and lactoferrin concentrations. In supplemented ewes, the level of colostral IgG was higher at T0 and a higher level of serum IgG was observed in lambs born from supplemented mothers and artificially fed, while no effect of supplementation was observed in the mothered lamb groups. Using a metabolomic approach, we showed that supplementation affected OS composition with significantly higher levels of colostral Neu-5Gc compounds up to 5 h after birth. No effect of supplementation was observed on bacterial composition. Our data suggest that live yeast supplementation offsets the negative impact of early separation and incomplete colostrum feeding in neonate lambs.
HAL (Le Centre pour la Communication Scientifique Directe), Oct 9, 2017
International audienceThe bovine gastrointestinal tract (GIT) is the main reservoir for enterohae... more International audienceThe bovine gastrointestinal tract (GIT) is the main reservoir for enterohaemorrhagic Escherichia coli (EHEC) responsible for food-borne infections. Therefore, it is crucial to develop strategies, such as EHEC suppression by antagonistic microorganisms, to reduce EHEC survival in the GIT of cattle and to limit shedding and food contamination. Most human-derived Lactobacillus reuteri strains produce hydroxypropionaldehyde (HPA), an antimicrobial compound, during anaerobic reduction of glycerol. The capacity of L. reuteri LB1-7 to produce HPA and its antimicrobial activity against EHEC FCH6 were evaluated in bovine rumen fluid (RF) under strict anaerobiosis. EHEC was totally suppressed when incubated in RF inoculated with L. reuteri LB1-7 and supplemented with 80 mM glycerol. The addition of LB1-7 or glycerol alone did not modify EHEC survival in RF. Glycerol was converted to HPA (up to 14 mM) and lactate (30 mM) by LB1-7, but only HPA production seemed to be responsible for EHEC suppression. Furthermore, L. reuteri LB1-7 did not have major adverse effects on ruminal digestion of common feedstuffs. The bactericidal activity of L. reuteri LB1-7, the concentration of glycerol required and the level of HPA secreted depended on physiological and ecological environments. In vitro experiments also showed that EHEC inoculated in rumen fluid and exposed to L. reuteri and glycerol had a very limited growth in rectal contents
Background Artificial rearing system, commonly used in prolific sheep breeds, is associated to in... more Background Artificial rearing system, commonly used in prolific sheep breeds, is associated to increased mortality and morbidity rates before weaning, which might be linked to perturbations in digestive tract maturation, including microbiota colonization. This study evaluated the effect of rearing mode (mothered or artificially reared) on the establishment of the rumen and intestinal microbiome of lambs from birth to weaning. We also measured immunological and zootechnical parameters to assess lambs' growth and health. GIT anatomy as well as rumen and intestinal epithelium gene expression were also analysed on weaned animals to assess possible long-term effects of the rearing practice. Results Total VFA concentrations were higher in mothered lambs at 2 months of age, while artificially-reared lambs had lower average daily gain, a more degraded sanitary status and lower serum IgG concentration in the early growth phase. Metataxonomic analysis revealed higher richness of bacterial and eukaryote populations in mothered vs. artificially-reared lambs in both Rumen and Feces. Beta diversity analysis indicated an evolution of rumen and fecal bacterial communities in mothered lambs with age, not observed in artificially-reared lambs. Important functional microorganisms such as the cellulolytic bacterium Fibrobacter succinogenes and rumen protozoa did not establish correctly before weaning in artificially-reared lambs. Enterobacteriaceae and Escherichia coli were dominant in the fecal microbiota of mothered lambs, but main E. coli virulence genes were not found differential between the two groups, suggesting they are commensal bacteria which could exert a protective effect against pathogens. The fecal microbiota of artificially-reared lambs had a high proportion of lactic acid bacteria taxa. No difference was observed in mucosa gene expression in the two lamb groups after weaning.
HAL (Le Centre pour la Communication Scientifique Directe), Oct 13, 2021
International audienceArtificial rearing system is commonly used in prolific meat breeds of sheep... more International audienceArtificial rearing system is commonly used in prolific meat breeds of sheep or in dairy breeds of goats. However, it represents a welfare issue due to increased mortality rates and morbidity. In this study we compared 14 lambs kept with their dams until weaning (mothered=MOT) to their twin counterparts separated 12h after birth from their mothers and reared artificially (ART), to better understand how the rearing mode, and thus the maternal presence, affect GIT microbiome, immunocompetence, health status and performance of lambs until two weeks postweaning.Rumen and fecal microbiota establishment was monitored with qPCR quantification of targeted populations and 16S and 18S amplicon sequencing. Rumen fermentative parameters and serum IgG and sanitary status were measured. Growth was recorded until 2 weeks postweaning. Then, 5 lambs per group were slaughtered, the GIT segments were weighted, and rumen papillae measurements were performed. In rumen and colon tissues, expression of targeted genes encoding for immune function, metabolic activity, and epithelium integrity was measured by RT-qPCR.Early separation with the dam clearly affected digestive microbiota establishment, with a strong delay in colonization by key members of rumen microbiome in ART vs MOT lambs. Fecal concentrations of lactic acid producing bacteria were higher in ART than in MOT in the first weeks of life suggesting differences in fermentation capacities in the hindgut, maybe associated to dysbiosis. A limited immune passive transfer was observed in ART compared to MOT lambs. ART lambs had lower ADG in early phase but finally caught up postweaning. Little or no changes in GIT anatomy, rumen papillae development, or epithelium gene expression were measured at slaughter. The presence of the mother and rearing mode influence GIT microbiota and health associated parameters in offspring in early life and the first month of life is a crucial period in artificial rearing system
HAL (Le Centre pour la Communication Scientifique Directe), Oct 13, 2021
International audienceEnterotoxigenic Escherichia coli (ETEC) is the principal pathogen responsib... more International audienceEnterotoxigenic Escherichia coli (ETEC) is the principal pathogen responsible for post-weaning diarrhea in newly weaned piglets. Expansion of ETEC at weaning is thought to be the consequence of various stress factors such as transient anorexia, dietary change or increase in intestinal inflammation and permeability, but the exact mechanisms remain to be elucidated. As the use of animal experiments raise more and more ethical concerns, we used a recently developed in vitro model of piglet colonic microbiome and mucobiome, the MPigut-IVM, to evaluate the effects of a simulated weaning transition and pathogen challenge at weaning. Our data suggested that the tested factors impacted the composition and functionality of the MPigut-IVM microbiota. The simulation of weaning transition led to an increase in relative abundance of the Prevotellaceae family, which was further promoted by the presence of the ETEC strain. In contrast, several beneficial families such as Bacteroidiaceae or Ruminococcaceae and gut health related short chain fatty acids like butyrate or acetate were reduced upon simulated weaning. Moreover, the incubation of MPigut-IVM filtrated effluents with porcine intestinal cell cultures showed that ETEC challenge in the in vitro model led to an increased expression of pro-inflammatory genes by the porcine cells. This study provides insights about the etiology of a dysbiotic microbiota in post-weaning piglets and showed that in vitro MPigut-IVM can be efficiently coupled to porcine epithelial cells to study host-microbiota crosstalk.P075 Periparturient Holstein cows of varying subacute ruminal acidosis phenotype differ in rumen bacterial community an
bioRxiv (Cold Spring Harbor Laboratory), Oct 15, 2021
Colostrum quality is of paramount importance in the management of optimal ruminant growth and inf... more Colostrum quality is of paramount importance in the management of optimal ruminant growth and infectious disease prevention in early life. Live yeast supplementation effect during the last month of gestation was evaluated on ewes' colostrum composition. Two groups of ewes (n=14) carrying twin lambs were constituted and twins were separated into groups (mothered or artificially-fed) 12h after birth. Nutrient, oligosaccharides (OS), IgG and lactoferrin concentrations were measured over 72h after lambing, and bacterial community was described in colostrum collected at parturition (T0). Immune passive transfer was evaluated through IgG measurement in lamb serum. In both groups, colostral nutrient, OS concentrations and IgG concentrations in colostrum and lamb serum decreased over time, (p < 0.01) except for lactose, which slightly increased (p < 0.001) and lactoferrin which remained stable. Bacterial population was stable over time with high relative abundances of Aerococcaceae, Corynebacteriaceae, Moraxellaceae and Staphylococcaceae in T0-colostrum. No effect of supplementation was observed in nutrient and lactoferrin concentrations. In supplemented ewes, colostral IgG level was higher at T0 and a higher level of serum IgG was observed in lambs born from supplemented mothers and artificially-fed, while no effect of supplementation was observed in the mothered lambs groups. Using a metabolomic approach, we showed that supplementation affected OS composition with significantly higher levels of colostral Neu-5Gc compounds up to 5h after birth. No effect of supplementation was observed on bacterial composition. Our data suggest that live yeast supplementation offsets the negative impact of early separation and incomplete colostrum feeding in neonate lambs.
Colostrum quality is of paramount importance in the management of optimal ruminant growth and inf... more Colostrum quality is of paramount importance in the management of optimal ruminant growth and infectious disease prevention in early life. Live yeast supplementation effect during the last month of gestation was evaluated on ewes' colostrum composition. Two groups of ewes (n = 14) carrying twin lambs were constituted and twins were separated into groups (mothered or artificially fed) 12 h after birth. Nutrient, oligosaccharides (OS), IgG and lactoferrin concentrations were measured over 72 h after lambing, and bacterial community was described in colostrum collected at parturition (T0). Immune passive transfer was evaluated through IgG measurement in lamb serum. In both groups, colostral nutrient, OS concentrations and IgG concentrations in colostrum and lamb serum decreased over time (P < 0⋅01), except for lactose, which slightly increased (P < 0⋅001), and lactoferrin, which remained stable. Bacterial population was stable over time with high relative abundances of Aerococcaceae, Corynebacteriaceae, Moraxellaceae and Staphylococcaceae in T0 colostrum. No effect of supplementation was observed in nutrient and lactoferrin concentrations. In supplemented ewes, the level of colostral IgG was higher at T0 and a higher level of serum IgG was observed in lambs born from supplemented mothers and artificially fed, while no effect of supplementation was observed in the mothered lamb groups. Using a metabolomic approach, we showed that supplementation affected OS composition with significantly higher levels of colostral Neu-5Gc compounds up to 5 h after birth. No effect of supplementation was observed on bacterial composition. Our data suggest that live yeast supplementation offsets the negative impact of early separation and incomplete colostrum feeding in neonate lambs.
HAL (Le Centre pour la Communication Scientifique Directe), Oct 9, 2017
International audienceThe bovine gastrointestinal tract (GIT) is the main reservoir for enterohae... more International audienceThe bovine gastrointestinal tract (GIT) is the main reservoir for enterohaemorrhagic Escherichia coli (EHEC) responsible for food-borne infections. Therefore, it is crucial to develop strategies, such as EHEC suppression by antagonistic microorganisms, to reduce EHEC survival in the GIT of cattle and to limit shedding and food contamination. Most human-derived Lactobacillus reuteri strains produce hydroxypropionaldehyde (HPA), an antimicrobial compound, during anaerobic reduction of glycerol. The capacity of L. reuteri LB1-7 to produce HPA and its antimicrobial activity against EHEC FCH6 were evaluated in bovine rumen fluid (RF) under strict anaerobiosis. EHEC was totally suppressed when incubated in RF inoculated with L. reuteri LB1-7 and supplemented with 80 mM glycerol. The addition of LB1-7 or glycerol alone did not modify EHEC survival in RF. Glycerol was converted to HPA (up to 14 mM) and lactate (30 mM) by LB1-7, but only HPA production seemed to be responsible for EHEC suppression. Furthermore, L. reuteri LB1-7 did not have major adverse effects on ruminal digestion of common feedstuffs. The bactericidal activity of L. reuteri LB1-7, the concentration of glycerol required and the level of HPA secreted depended on physiological and ecological environments. In vitro experiments also showed that EHEC inoculated in rumen fluid and exposed to L. reuteri and glycerol had a very limited growth in rectal contents
Background Artificial rearing system, commonly used in prolific sheep breeds, is associated to in... more Background Artificial rearing system, commonly used in prolific sheep breeds, is associated to increased mortality and morbidity rates before weaning, which might be linked to perturbations in digestive tract maturation, including microbiota colonization. This study evaluated the effect of rearing mode (mothered or artificially reared) on the establishment of the rumen and intestinal microbiome of lambs from birth to weaning. We also measured immunological and zootechnical parameters to assess lambs' growth and health. GIT anatomy as well as rumen and intestinal epithelium gene expression were also analysed on weaned animals to assess possible long-term effects of the rearing practice. Results Total VFA concentrations were higher in mothered lambs at 2 months of age, while artificially-reared lambs had lower average daily gain, a more degraded sanitary status and lower serum IgG concentration in the early growth phase. Metataxonomic analysis revealed higher richness of bacterial and eukaryote populations in mothered vs. artificially-reared lambs in both Rumen and Feces. Beta diversity analysis indicated an evolution of rumen and fecal bacterial communities in mothered lambs with age, not observed in artificially-reared lambs. Important functional microorganisms such as the cellulolytic bacterium Fibrobacter succinogenes and rumen protozoa did not establish correctly before weaning in artificially-reared lambs. Enterobacteriaceae and Escherichia coli were dominant in the fecal microbiota of mothered lambs, but main E. coli virulence genes were not found differential between the two groups, suggesting they are commensal bacteria which could exert a protective effect against pathogens. The fecal microbiota of artificially-reared lambs had a high proportion of lactic acid bacteria taxa. No difference was observed in mucosa gene expression in the two lamb groups after weaning.
HAL (Le Centre pour la Communication Scientifique Directe), Oct 13, 2021
International audienceArtificial rearing system is commonly used in prolific meat breeds of sheep... more International audienceArtificial rearing system is commonly used in prolific meat breeds of sheep or in dairy breeds of goats. However, it represents a welfare issue due to increased mortality rates and morbidity. In this study we compared 14 lambs kept with their dams until weaning (mothered=MOT) to their twin counterparts separated 12h after birth from their mothers and reared artificially (ART), to better understand how the rearing mode, and thus the maternal presence, affect GIT microbiome, immunocompetence, health status and performance of lambs until two weeks postweaning.Rumen and fecal microbiota establishment was monitored with qPCR quantification of targeted populations and 16S and 18S amplicon sequencing. Rumen fermentative parameters and serum IgG and sanitary status were measured. Growth was recorded until 2 weeks postweaning. Then, 5 lambs per group were slaughtered, the GIT segments were weighted, and rumen papillae measurements were performed. In rumen and colon tissues, expression of targeted genes encoding for immune function, metabolic activity, and epithelium integrity was measured by RT-qPCR.Early separation with the dam clearly affected digestive microbiota establishment, with a strong delay in colonization by key members of rumen microbiome in ART vs MOT lambs. Fecal concentrations of lactic acid producing bacteria were higher in ART than in MOT in the first weeks of life suggesting differences in fermentation capacities in the hindgut, maybe associated to dysbiosis. A limited immune passive transfer was observed in ART compared to MOT lambs. ART lambs had lower ADG in early phase but finally caught up postweaning. Little or no changes in GIT anatomy, rumen papillae development, or epithelium gene expression were measured at slaughter. The presence of the mother and rearing mode influence GIT microbiota and health associated parameters in offspring in early life and the first month of life is a crucial period in artificial rearing system
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