Project description:In this study, samples of 16 dairy cows from a MAP infected farm were used. Serum, milk and fecal samples were collected. Categorizing these cows into two groups based on their MAP infection status different standard methods for detection MAP were applied. Healthy controls showed no positive results in enzyme-linked immunosorbent assay (ELISA) with serum and milk samples (cattletype MAP Ab, Qiagen, Hilden, Germany; In-direct, IDVet, Grabels, France) and after cultivation of fecal samples on commercial Her-rold´s Egg Yolk Agars (HEYM agar, Becton Dickinson, Heidelberg, Germany) for 12 weeks. Cows with positive results were grouped into MAP infected cows. Specifically, for mass spectrometry analysis serum of seven MAP infected cows and seven healthy controls were used. All animals were from the same farm and were kept under the same environmental conditions. For additional mass spectrometry analysis with a further control group sam-ples of 21 dairy cows from an uninfected farm were examined. All cattle from this farm showed negative results in ELISA with serum and milk samples. Additionally, there was never a positive result in regularly tested fecal samples and sock swab samples of this farm. For verification of differential CTSS expression in Western blot analysis five dairy cows from another infected farm were consultedincluded. MAP status of these cows was analyzed by cultivation of fecal samples on HEYM agar for 12 weeks and ELISA with se-rum samples. In detail, two cattle were categorized into healthy controls and three cattle into MAP infected cows. Withdrawal of bovine venous whole blood and experi-mental protocols were approved by the local authority, Government of Upper Bavaria, permit no. ROB-55.2-2532.Vet_03-17-106.
Project description:Maternal secretor status is one of the determinants of human milk oligosaccharides (HMOs) composition, which in turn changes the gut microbiota composition of infants. To understand if this change in gut microbiota impacts immune cell composition, intestinal morphology and gene expression, day 21-old germ-free mice were transplanted with fecal microbiota from infants whose mothers were either secretors (SMM) or non-secretors (NSM) or from infants consuming dairy-based formula (MFM). For each group, one set of mice was supplemented with HMOs. HMO supplementation did not significantly impact the microbiota diversity however, SMM mice had higher abundance of genus Bacteroides, Bifidobacterium, and Blautia, whereas, in the NSM group, there were higher abundance of Akkermansia, Enterocloster, and Klebsiella. In MFM, gut microbiota was represented mainly by Parabacteroides, Ruminococcaceae_unclassified, and Clostrodium_sensu_stricto. In mesenteric lymph node, Foxp3+ T cells and innate lymphoid cells type 2 (ILC2) were increased in MFM mice supplemented with HMOs while in the spleen, they were increased in SMM+HMOs mice. Similarly, serum immunoglobulin A (IgA) was also elevated in MFM+HMOs group. Distinct global gene expression of the gut was observed in each microbiota group, which was enhanced with HMOs supplementation. Overall, our data shows that distinct infant gut microbiota due to maternal secretor status or consumption of dairy-based formula and HMO supplementation impacts immune cell composition, antibody response and intestinal gene expression in a mouse model.
Project description:The consumption of fermented food has been linked to positive health outcomes due to a variety of functional properties. Fermented dairy constitutes a major dietary source and contains lactoseas main carbohydrate and living starter cultures. To investigate whether nutritional and microbial modulation impacted intestinal microbiota composition and activity, we employed fecal microbiota fermentations and a dairy model system consisting of lactose and β-galactosidase positive and negative Streptococcus thermophilus. Based on 16S rRNA gene based microbial community analysis, we observed that lactose addition increased the abundance of Bifidobacteriaceae, and of Veillonellaceae and Enterobacteraceae in selected samples. The supplied lactose was hydrolysed within 24 h of fermentation and led to higher expression of community indigenous β-galactosidases. Targeted protein analysis confirmed that bifidobacteria contributed most β-galactosidases together with other taxa including Escherichia coli and Anaerobutyricum hallii. Lactose addition led to 1.1-1.8 fold higher levels of butyrate compared to controls likely due to (i) lactate-crossfeeding and (ii) direct lactose metabolism by butyrate producing Anaerobutyricum and Faecalibacterium spp. Representatives of both genera used lactose to produce butyrate in single cultures. When supplemented at around 5.5 log cells mL-1, S. thermophilus or its beta-galactosidase negative mutant outnumbered the indigenous Streptococcaceae population at the beginning of fermentation but had no impact on lactose utilisation and final SCFA profiles. This study brings forward new fundamental insight into interactions of major constituents of fermented dairy with the intestinal microbiota. We provide evidence that lactose addition increases fecal microbiota production of butyrate through cross-feeding and direct metabolism without contribution of starter cultures.
Project description:The transition period is the most critical stage in the lactation cycle of dairy cattle. During this period, cows are subjected to high levels of oxidative stress. One way of managing this stress is through mineral supplementation with antioxidant micronutrients. The aim of this study was to evaluate the gene expression of transition dairy cows supplemented with the antioxidant trace elements copper (Cu), zinc (Zn), manganese (Mn) and selenium (Se). The study was carried out in a commercial Holstein dairy farm located in General Belgrano, province of Buenos Aires, Argentina. Cows (n=200) were randomly assigned to either a supplemented or a control group. Blood samples were obtained seven days after calving and used to determine superoxide dismutase and glutathione peroxidase activity, antioxidant capacity and thiobarbituric acid reactive substances. Additionally, RNA-sequencing analysis was performed. The oxidative stress index differed significantly between groups, despite only two differentially expressed genes which codify for second messengers (adjusted p value < 0.05). This would suggest that trace mineral supplementation of transition dairy cows would not induce changes in gene expression profiles in pathways associated with oxidative stress and immune function, since their expression is already high in response to the high oxidative stress levels and the dietary changes associated with this period. Nevertheless, considering the role of these minerals as cofactors, a higher availability in the supplemented group would increase antioxidant enzyme activity.
Project description:Fatty liver disease is prevalent during parturition in dairy cattle. Therefore, there is an urgent need to develop novel, sensitive biomarkers for the early diagnosis of the metabolic disorders. Macroproteomics revealed that the fecal microbial community changes significantly when animal develops fatty liver disease. The microbial changes in cows with severe fatty liver (SFL) were greater than cows with moderate fatty liver (MFL) and normal condition (Norm). This suggests that microorganisms play an important role in the pathogenesis of metabolic disorders. In this study, feces-sourced microorganisms and microbial proteins were identified and testified as novel biomarkers for the early diagnosis of fatty liver disease in cattle. For example, the AUC (area under curve) values, based on Receiver Operating Characteristics analysis, of using the combination of Lachnoanaerobaculum and Bifidobacterium (at the genus level) to discriminate MFL and SFL animals reached 0.944 and 0.867, respectively. and 0.922 and 0.985, respectively, for the combination of Bifidobacterium pseudolongum and Lachnospiraceae bacterium (at the species level). Interestingly, the differentially expressed microbial proteins are closely related to the identified microorganisms. For example, the majority of the top 20 microbial proteins with significant expression differences were derived from Bifidobacterium pseudolongum. Bifidobacterium pseudolongum was considered a prominent potential biomarker for the diagnosis of metabolic disorders, especially in fatty liver cattle. The results of this study confirm that microbial signatures have a causal contribution to the pathophysiological mechanism of non-alcoholic fatty liver disease (NAFLD), but also shed light on fecal microbiota transfer (FMT) experiments in treating NAFLD.
Project description:miRNA profiling of bovine satellite cells (BSC) differentiated into myotubes (6th day of in vitro differentiation). BSC isolated from m. semitendinosus of beef (Hereford & Limousine) and dairy (Holstein-Friesian) cattle. Goal was to determine differences in miRNA expresion during in vitro myogenesis in beef vs dairy cattle used as a control.
Project description:Analysis of breast cancer survivors' gut microbiota after lifestyle intervention, during the COVID-19 lockdown, by 16S sequencing of fecal samples.
Project description:We found that low protein diet consumption resulted in decrease in the percentage of normal Paneth cell population in wild type mice, indicating that low protein diet could negatively affect Paneth cell function. We performed fecal microbiota composition profiling. Male mice were used at 4-5 weeks of age. Fecal samples were collected for microbiome analysis.
Project description:This study aimed to analyze changes in gut microbiota composition in mice after transplantation of fecal microbiota (FMT, N = 6) from the feces of NSCLC patients by analyzing fecal content using 16S rRNA sequencing, 10 days after transplantation. Specific-pathogen-free (SPF) mice were used for each experiments (N=4) as controls.