Agriculture is at the pivot point between anthroposphere, biosphere, and atmosphere. Innovative solutions are needed to reduce agricultural emissions and improve sustainability. Microalgae animal feed could be such a solution. This study aimed to evaluate the effects of 10 freshwater microalgae: Auxenochlorella protothecoides, Chlamydomonas pulvinate, Chlorella luteoviridis, Chlorella variabilis, Euglena mutabilis, Parachlorella kessleri, Stichococcus bacillaris, Tetradesmus acuminatus, Tetradesmus obliquus, and Tetraselmis gracilis, on ruminal methane (CH4) production, nutrient digestibility, and rumen fermentation using the in vitro Hohenheim gas test. The microalgae were cultured in a carbon dioxide (CO2) incubator at 2% CO2, at the optimal conditions for each strain. The highest producers were P. kessleri and T. obliquus, with a biomass concentration of 0.69 and 0.73 g/L·d, respectively. Their PUFA contents ranged from 33.2% to 69.1% of total fatty acids. Microalgae were tested at a 15% replacement in a control basal diet of 40.0% DM grass silage, 40.0% maize silage, 15% hay, and 5% concentrate. Data were analyzed using a mixed model in R. Ruminal CH4 production was reduced by 15.4%, 17.4%, and 16.4% in diets containing A. protothecoides, C. luteoviridis, and P. kessleri, respectively, compared with the control diet. Similarly, these diets reduced in vitro organic matter digestibility by 3.5%, 5.2%, and 5.4%, respectively. However, only A. protothecoides reduced CH4/CO2 ratio by 3.5% compared with the control diet. Propionate molar proportion was decreased by 2.4, 3.0, 2.5, and 2.5 percentage points for diets containing Ch. pulvinate, E. mutabilis, P. kessleri, and T. obliquus, respectively. Marginal effects of dietary variables were analyzed using the generalized additive model framework, revealing a negative relationship between dietary PUFA, sulfur content, and CH4 production, and a negative relationship between dietary PUFA and CH4/CO2 ratio. Incorporating high-PUFA microalgae in ruminant diets shows potential for reducing enteric CH4 emissions, warranting further investigation.
- MeSH
- Rumen * metabolism MeSH
- Diet veterinary MeSH
- Fermentation MeSH
- Animal Feed MeSH
- Methane * metabolism MeSH
- Microalgae * metabolism MeSH
- Silage MeSH
- Cattle MeSH
- Fresh Water MeSH
- Digestion MeSH
- Animals MeSH
- Check Tag
- Cattle MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
BACKGROUND: Exclusive enteral nutrition (EEN) is an effective treatment for active Crohn's disease (CD). This study explored the immunostimulatory potential of a cell-free fecal filtrate and related this with changes in the fecal microbiota and metabolites in children with active CD undertaking treatment with EEN. METHODS: Production of tumor necrosis factor α (TNFα) from peripheral blood mononuclear cells was measured following their stimulation with cell-free fecal slurries from children with CD, before, during, and at completion of EEN. The metabolomic profile of the feces used was quantified using proton nuclear magnetic resonance and their microbiota composition with 16S ribosomal RNA sequencing. RESULTS: Following treatment with EEN, 8 (72%) of 11 patients demonstrated a reduction in fecal calprotectin (FC) >50% and were subsequently labeled FC responders. In this subgroup, TNFα production from peripheral blood mononuclear cells was reduced during EEN (P = .008) and reached levels like healthy control subjects. In parallel to these changes, the fecal concentrations of acetate, butyrate, propionate, choline, and uracil significantly decreased in FC responders, and p-cresol significantly increased. At EEN completion, TNFα production from peripheral blood mononuclear cells was positively correlated with butyrate (rho = 0.70; P = .016). Microbiota structure (β diversity) was influenced by EEN treatment, and a total of 28 microbial taxa changed significantly in fecal calprotectin responders. At EEN completion, TNFα production positively correlated with the abundance of fiber fermenters from Lachnospiraceae_UCG-004 and Faecalibacterium prausnitzii and negatively with Hungatella and Eisenbergiella tayi. CONCLUSIONS: This study offers proof-of concept data to suggest that the efficacy of EEN may result from modulation of diet-dependent microbes and their products that cause inflammation in patients with CD.
- MeSH
- Crohn Disease * therapy microbiology immunology MeSH
- Child MeSH
- Enteral Nutrition * methods MeSH
- Feces * microbiology chemistry MeSH
- Leukocyte L1 Antigen Complex * analysis MeSH
- Leukocytes, Mononuclear immunology metabolism MeSH
- Humans MeSH
- Adolescent MeSH
- Gastrointestinal Microbiome * MeSH
- Tumor Necrosis Factor-alpha * metabolism MeSH
- Check Tag
- Child MeSH
- Humans MeSH
- Adolescent MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Pichia pastoris, a methylotrophic yeast, is known to be an efficient host for heterologous proteins production. In this study, a recombinant P. pastoris Y11430 was found better for β-glucosidase activity in comparison with a wild type P. pastoris Y11430 strain, and thereby, subjected to methanol intermittent feed profiling for β-glucosidase production. The results showed that at 72 h of cultivation time, the cultures with 16.67% and 33.33% methanol feeding with constant rate could produce the total dry cell weight of 52.23 and 118.55 g/L, respectively, while the total mutant β-glucosidase activities were 1001.59 and 3259.82 units, respectively. The methanol feeding profile was kept at 33% with three methanol feeding strategies such as constant feed rate, linear feed rate, and exponential feed rate which were used in fed-batch fermentation. At 60 h of cultivation, the highest total mutant β-glucosidase activity was 2971.85 units for exponential feed rate culture. On the other hand, total mutant β-glucosidase activity of the constant feed rate culture and linear feed rate culture were 1682.25 and 1975.43 units, respectively. The kinetic parameters of exponential feed rate culture were specific growth rate on glycerol 0.228/h, specific growth of methanol 0.061/h, maximum total dry cell weight 196.73 g, yield coefficient biomass per methanol ([Formula: see text]) 0.57 gcell/gMeOH, methanol consumption rate ([Formula: see text]) 5.76 gMeOH/h, and enzyme productivity ([Formula: see text]) 75.96 units/h. In conclusion, higher cell mass and β- glucosidase activity were produced under exponential feed rate than constant and linear feed rates.
Previous studies indicated an intrinsic relationship between infant diet, intestinal microbiota composition and fermentation activity with a strong focus on the role of breastfeeding on microbiota composition. Yet, microbially formed short-chain fatty acids acetate, propionate and butyrate and other fermentation metabolites such as lactate not only act as substrate for bacterial cross-feeding and as mediators in microbe-host interactions but also confer antimicrobial activity, which has received considerably less attention in the past research. It was the aim of this study to investigate the nutritional-microbial interactions that contribute to the development of infant gut microbiota with a focus on human milk oligosaccharide (HMO) fermentation. Infant fecal microbiota composition, fermentation metabolites and milk composition were analyzed from 69 mother-infant pairs of the Swiss birth cohort Childhood AlleRgy nutrition and Environment (CARE) at three time points depending on breastfeeding status defined at the age of 4 months, using quantitative microbiota profiling, HPLC-RI and 1H-NMR. We conducted in vitro fermentations in the presence of HMO fermentation metabolites and determined the antimicrobial activity of lactate and acetate against major Clostridiaceae and Peptostreptococcaceae representatives. Our data show that fucosyllactose represented 90% of the HMOs present in breast milk at 1- and 3-months post-partum with fecal accumulation of fucose, 1,2-propanediol and lactate indicating fermentation of HMOs that is likely driven by Bifidobacterium. Concurrently, there was a significantly lower absolute abundance of Peptostreptococcaceae in feces of exclusively breastfed infants at 3 months. In vitro, lactate inhibited strains of Peptostreptococcaceae. Taken together, this study not only identified breastfeeding dependent fecal microbiota and metabolite profiles but suggests that HMO-derived fermentation metabolites might exert an inhibitory effect against selected gut microbes.
- Publication type
- Journal Article MeSH
Dahi, an artisanal fermented milk product, widely consumed in Pakistan, is microbiologically diverse, and many bacterial communities await investigation. The current study is first to present probiotic assessment of Bacillus species strains isolated from dahi. Based on 49 identified strains assessed, only 6 strains, i.e., Bacillus licheniformis QAUBL19, QAUBL1901, and QAUBL1902; Bacillus mycoides QAUBM19 and QAUBM1901; and Bacillus subtilis QAUBSS1 were having prominent persistence in the simulated gastrointestinal fluids, being non-hemolytic, with no DNase activity. Probiotic characteristics, cholesterol-assimilating, and carbohydrate-fermenting capabilities were assessed for all the strains. These six strains each showed variant cholesterol assimilating abilities. B. licheniformis QAUBL19 retaining most desired probiotic traits presented both notable cholesterol assimilating and bile salt hydrolase activities. It can be used as a probiotic of choice with hypocholesterolemia ability. B. subtilis QAUBSS1 showed wide carbohydrate fermentation ability and strongest antibacterial potential. It is likely to be considered a probiotic for living beings and starter culture for fermentation of food/feed.
- MeSH
- Bacillus * genetics MeSH
- Cholesterol MeSH
- Fermentation MeSH
- Cultured Milk Products * MeSH
- Milk microbiology MeSH
- Probiotics * MeSH
- Carbohydrates MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
Aging is a progressive, unalterable physiological degradation process of living organisms, which leads to deterioration of biological function and eventually to senescence. The most prevalent factor responsible for aging is the accumulation of damages resulting from oxidative stress and dysbiosis. D-galactose-induced aging has become a hot topic, and extensive research is being conducted in this area. Published literature has reported that the continuous administration of D-galactose leads to the deterioration of motor and cognitive skills, resembling symptoms of aging. Hence, this procedure is employed as a model for accelerated aging. This review aims to emphasize the effect of D-galactose on various bodily organs and underline the role of the Lactobacillus sp. in the aging process, along with its anti-oxidative potential. A critical consideration to the literature describing animal models that have used the Lactobacillus sp. in amending D-galactose-induced aging is also given. KEY POINTS: • D-Galactose induces the aging process via decreasing the respiratory chain enzyme activity as well as ATP synthesis, mitochondrial dysfunction, and increased ROS production. • D-Galactose induced aging primarily affects the brain, heart, lung, liver, kidney, and skin. • The anti-oxidative potential of Lactobacillus sp. in improving D-galactose-induced aging in animal models via direct feeding and feeding of Lactobacillus-fermented food.
- MeSH
- Antioxidants * metabolism MeSH
- Galactose * MeSH
- Lactobacillus metabolism MeSH
- Oxidative Stress MeSH
- Aging MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Zearalenone (ZEN) is a toxic secondary metabolite of Fusarium sp. commonly found in wheat, corn, and other crops. In addition to economic losses, ZEN can seriously endanger the health of both humans and livestock, thus presenting an urgent need for ZEN-detoxifying enzymes that function in the extreme heat or pH conditions of industrial fermenters. Here, we identify and characterize the activity of the ZEN-degrading enzyme from Exophiala spinifera, ZHD_LD, which shares 60.15% amino acid identity and a conserved catalytic triad with the well-characterized ZEN-detoxifying protein ZHD101 from Clonostachys rosea. Biochemical activity and stability assays indicated that purified recombinant ZHD_LD exhibited high activity against ZEN with optimal reaction conditions of 50 °C and pH 7.0-10.0. Structural modeling of the ZHD_LD active site and comparison with ZHD101 revealed its likely mechanism of ZEN degradation. This research provides an industrially valuable candidate enzyme for ZEN detoxification in food and livestock feed.
- MeSH
- Fusarium * metabolism MeSH
- Hydrolases metabolism MeSH
- Humans MeSH
- Triticum metabolism MeSH
- Zearalenone * chemistry metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
A batch culture technique was used to evaluate dry matter (DM) digestibility of corn stover (Zea mays L.) and kudzu, Pueraria montana (Lour.) Merr. after solid-state fermentation (SSF) with a white-rot fungus, Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm. Five dietary treatments consisting of mixtures of corn stover (C) and kudzu (K) in varying ratios, (1) 100C:0K, (2) 75C:25K, (3) 50C:50K, (4) 25C:75K, and (5) 0C:100K, were inoculated with P. ostreatus (MBFBL 400) and subjected to SSF for 0, 35, and 77 days. The study was arranged as a 5 × 3 factorial design with 3 replicates. Grass hay was included in the study as a control. Interactions (P < 0.05) between treatments and fermentation time were noted for the fermentation kinetics. Asymptotic gas was the highest (P < 0.05) for 0C:100K and 100C:0K on day 77. Treatment effect (P < 0.001) and treatment × fermentation time interaction (P < 0.001) were noted for in vitro dry matter digestibility (IVDMD). On day 77, treatment 4 had the highest (P < 0.001) IVDMD value, while treatment 1 had the lowest (P < 0.001) IVDMD. There was no difference (P > 0.05) between treatments 3, 5, and control. Numerically, the ranking of their IVDMD values from the highest to the lowest is 4 > 2 > 5 > control >3 > 1. The results show that the treated corn stover and kudzu mixes were comparable with the control, which is good quality hay. This is the first report that demonstrates the potential use of a combined mixture of corn stover and kudzu in ruminant animal feed development.
- MeSH
- Biomass MeSH
- Fermentation MeSH
- Kinetics MeSH
- Animal Feed analysis microbiology MeSH
- Zea mays chemistry microbiology MeSH
- Nutritive Value * MeSH
- Pleurotus metabolism MeSH
- Gases metabolism MeSH
- Pueraria chemistry microbiology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
Pediocin PA-1 is a bacteriocin that shows strongly anti-microbial activity against some Gram-positive pathogens such as Listeria monocytogenes, Staphylococcus aureus, and Enterococcus faecalis. With the broad inhibitory spectrum as well as high-temperature stability, pediocin has a potential application in the food preservation and pharmaceutical industry. Pediocin has been studied to express in many heterologous expression systems such as Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris as a free peptide. Here we showed in this study a new strategy by using yeast surface display system to produce the anchored pediocin PA-1 on the cell surface of Saccharomyces cerevisiae, which could be used directly as a pediocin resource. We had successfully constructed a recombinant S. cerevisiae W303 strain that could express pediocin PA-1 on the cell surface. The pediocin-expressing yeast could inhibit the growth of Shigella boydii and Shigella flexneri, which have never been reported before for pediocin activity. Besides, the pediocin expression level of the recombinant S. cerevisiae strain was also evaluated in three different media: synthetic defined (SD), basic medium (BM), and fermentation medium (FM). BM medium was shown to give the highest production yield of the recombinant yeast (4.75 ± 0.75 g dry cell weight per 1 L of culture) with the ratio number of the pediocin-expressing cells of 93.46 ± 2.45%. Taken together, the results clearly showed that pediocin can be displayed on yeast cell surface as anchored protein. The application of yeast cell surface system enables a new door of pediocin application on either food or feed industries. Graphical abstract.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Bacteria drug effects MeSH
- Bacteriocins MeSH
- Fermentation MeSH
- Genetic Vectors MeSH
- Microbial Sensitivity Tests MeSH
- Pediocins genetics metabolism pharmacology MeSH
- Recombinant Fusion Proteins genetics metabolism MeSH
- Saccharomyces cerevisiae genetics metabolism MeSH
- Publication type
- Journal Article MeSH
Fungi can improve stover digestibility due to their ability to secrete oxidative enzymes that depolymerize lignin, allowing the rumen microorganisms to access the polysaccharides of the plant cell wall. Some ascomycetes have shown good delignification capability; however, they have been scarcely evaluated for their ability to improve corn stover (CS) ruminal digestibility. We evaluated the laccase induction by CS of the CMU-196 strain of the ascomycete fungus Didymosphaeria sp. (syn. = Paraconiothyrium sp.). Also, we analyzed the capacity of such strain to modify the cell wall of CS and to improve its digestion by the ruminal microbiota. The CMU-196 strain showed a maximum extracellular laccase activity of 39.74 ± 0.24 U/L when an aqueous stover extract (SE, 10% v/v) was added to the growth medium. The addition of ground stover (GS, 2% w/v) increased the activity to a maximum of 262.27 ± 0.58 U/L. In solid-state fermentation (SSF) assays of GS, the strain degrades cell walls, destabilizing the vessels and tracheids of plant biomass; the protein content reaches a maximum of 33.2 g/kg dry matter (DM) at 70 days, while the crude fiber content shows the highest level of 314 g/kg DM at 14 days. SSF treatment of the CS increased the in vitro ruminal production of gas in a fraction that was considered nondigestible at 18 h, and gas production increased by 14% with respect to the untreated GS at 14 days. The CMU-196 strain can digest the plant cell wall and improve ruminal CS digestibility at a level equivalent to several basidiomycete species.
- MeSH
- Ascomycota enzymology growth & development metabolism MeSH
- Rumen microbiology MeSH
- Biomass MeSH
- Cell Wall metabolism ultrastructure MeSH
- Fermentation MeSH
- Animal Feed analysis microbiology MeSH
- Zea mays metabolism ultrastructure MeSH
- Laccase metabolism MeSH
- Lignin metabolism MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH