Lactic acid bacteria reduce bacterial diarrhea in rabbits via enhancing immune function and restoring intestinal microbiota homeostasis.

BACKGROUND: Numerous previous reports have demonstrated the efficacy of Lactic acid bacteria (LAB) in promoting growth and preventing disease in animals. In this study, Enterococcus faecium ZJUIDS-R1 and Ligilactobaciiius animalis ZJUIDS-R2 were isolated from the feces of healthy rabbits, and both strains showed good probiotic properties in vitro. Two strains (108CFU/ml/kg/day) were fed to weaned rabbits for 21 days, after which specific bacterial infection was induced to investigate the effects of the strains on bacterial diarrhea in the rabbits. RESULTS: Our data showed that Enterococcus faecium ZJUIDS-R1 and Ligilactobaciiius animalis ZJUIDS-R2 interventions reduced the incidence of diarrhea and systemic inflammatory response, alleviated intestinal damage and increased antibody levels in animals. In addition, Enterococcus faecium ZJUIDS-R1 restored the flora abundance of Ruminococcaceae1. Ligilactobaciiius animalis ZJUIDS-R2 up-regulated the flora abundance of Adlercreutzia and Candidatus Saccharimonas. Both down-regulated the flora abundance of Shuttleworthia and Barnesiella to restore intestinal flora balance, thereby increasing intestinal short-chain fatty acid content. CONCLUSIONS: These findings suggest that Enterococcus faecium ZJUIDS-R1 and Ligilactobaciiius animalis ZJUIDS-R2 were able to improve intestinal immunity, produce organic acids and regulate the balance of intestinal flora to enhance disease resistance and alleviate diarrhea-related diseases in weanling rabbits.

Probiotic characteristics of Lactobacillus plantarum E680 and its effect on Hypercholesterolemic mice.

BACKGROUND: Probiotics have been reported to reduce total cholesterol levels in vitro, but more evidence is needed to determine the clinical relevance of this activity. Chinese traditional fermented pickles are a good source of lactic acid bacteria. Therefore, pickle samples were collected for screening lactic acid bacteria based on their ability to survive stresses encountered during gastrointestinal passage and cholesterol reducing potency. RESULTS: Seventy five lactic acid bacteria strains were isolated from 22 fermented pickles. From these bacteria, Lactobacillus plantarum E680, showed the highest acid (85.25%) and bile tolerance (80.79%). It was sensitive to five of the eight antibiotics tested, inhibited the growth of four pathogenic bacteria, and reduced the total cholesterol level by 66.84% in broth culture. In vivo testing using hypercholesterolemic mice fed high-fat emulsion, independent of food intake, found that L. plantarum E680 suppressed body weight gain and reduced total cholesterol and low-density lipoprotein cholesterol levels, with no effect on high-density lipoprotein cholesterol. CONCLUSIONS: Chinese traditional fermented pickles are a good source for probiotics. L. plantarum E680, isolated from pickles, was acid and bile tolerant, sensitive to antibiotics, and reduced cholesterol levels both in vitro and in vivo. Based on these results, L. plantarum E680 may have potential as a novel probiotic for the development of cholesterol-lowering functional food.

Characterization of the indigenous microflora in raw and pasteurized buffalo milk during storage at refrigeration temperature by high-throughput sequencing.

The effect of refrigeration on bacterial communities within raw and pasteurized buffalo milk was studied using high-throughput sequencing. High-quality samples of raw buffalo milk were obtained from 3 dairy farms in the Guangxi province in southern China. Five liters of each milk sample were pasteurized (72°C; 15 s); and both raw and pasteurized milks were stored at refrigeration temperature (1-4°C) for various times with their microbial communities characterized using the Illumina Miseq platform (Novogene, Beijing, China). Results showed that both raw and pasteurized milks contained a diverse microbial population and that the populations changed over time during storage. In raw buffalo milk, Lactococcus and Streptococcus dominated the population within the first 24h; however, when stored for up to 72h the dominant bacteria were members of the Pseudomonas and Acinetobacter genera, totaling more than 60% of the community. In pasteurized buffalo milk, the microbial population shifted from a Lactococcus-dominated community (7d), to one containing more than 84% Paenibacillus by 21d of storage. To increase the shelf-life of buffalo milk and its products, raw milk needs to be refrigerated immediately after milking and throughout transport, and should be monitored for the presence of Paenibacillus. Results from this study suggest pasteurization should be performed within 24h of raw milk collection, when the number of psychrotrophic bacteria are low; however, as Paenibacillus spores are resistant to pasteurization, additional antimicrobial treatments may be required to extend shelf-life. The findings from this study are expected to aid in improving the quality and safety of raw and pasteurized buffalo milk.

Effect of heat and homogenization on in vitro digestion of milk.

Central to commercial fluid milk processing is the use of high temperature, short time (HTST) pasteurization to ensure the safety and quality of milk, and homogenization to prevent creaming of fat-containing milk. Ultra-high-temperature sterilization is also applied to milk and is typically used to extend the shelf life of refrigerated, specialty milk products or to provide shelf-stable milk. The structures of the milk proteins and lipids are affected by processing but little information is available on the effects of the individual processes or sequences of processes on digestibility. In this study, raw whole milk was subjected to homogenization, HTST pasteurization, and homogenization followed by HTST or UHT processing. Raw skim milk was subjected to the same heating regimens. In vitro gastrointestinal digestion using a fasting model was then used to detect the processing-induced changes in the proteins and lipids. Using sodium dodecyl sulfate-PAGE, gastric pepsin digestion of the milk samples showed rapid elimination of the casein and α-lactalbumin bands, persistence of the ß-lactoglobulin bands, and appearance of casein and whey peptide bands. The bands for ß-lactoglobulin were eliminated within the first 15min of intestinal pancreatin digestion. The remaining proteins and peptides of raw, HTST, and UHT skim samples were digested rapidly within the first 15min of intestinal digestion, but intestinal digestion of raw and HTST pasteurized whole milk showed some persistence of the peptides throughout digestion. The availability of more lipid droplets upon homogenization, with greater surface area available for interaction with the peptides, led to persistence of the smaller peptide bands and thus slower intestinal digestion when followed by HTST pasteurization but not by UHT processing, in which the denatured proteins may be more accessible to the digestive enzymes. Homogenization and heat processing also affected the ζ-potential and free fatty acid release during intestinal digestion. Stearic and oleic acids were broken down faster than other fatty acids due to their positions on the outside of the triglyceride molecule. Five different casein phosphopeptide sequences were observed after gastric digestion, and 31 sequences were found after intestinal digestion, with activities yet to be explored. Processing affects milk structure and thus digestion and is an important factor to consider in design of foods that affect health and nutrition.

Separation and quantification of milk casein from different buffalo breeds.

Understanding the milk protein expression profile in different buffalo breeds plays an important role in improving hybrid selection and determining the effects on milk protein synthesis. The aim of this research is to compare the differences in milk protein content, composition and distribution between River buffalo and their crossbreeds for hybrid screening. Four groups of milk samples that included Nili-Ravi (N), Murrah (M), a Nili-Ravi-Murrah crossbreed (M-N), and a crossbreed of river buffalo with local swamp buffalo (C) were collected. The protein composition of the buffalo milk was determined by RP-HPLC. A gel-based proteomic approach consisting of two-dimensional gel electrophoresis coupled with mass spectrometry was utilised for the detailed protein characterisation of milk from different breeds. The results of this analysis showed that the river/swamp buffalo crossbreed (C) displayed the highest content of total protein (4·46%) and κ-casein (11·14%) but the lowest content of α-lactalbumin (6·79%). By selecting 23 different protein spots among the four types of milk that contained the most spots corresponding to κ-casein, ß-casein and αs1-casein, correlations between the crossbreeds, protein polymorphism and phosphorylation could be made. The results of this study indicate that crossbreeding a swamp buffalo with a river buffalo has a notable effect on the protein content and composition that may be exploited for producing high-quality raw milk in food technology applications and dairy food production.

Effect of homogenization and pasteurization on the structure and stability of whey protein in milk.

The effect of homogenization alone or in combination with high-temperature, short-time (HTST) pasteurization or UHT processing on the whey fraction of milk was investigated using highly sensitive spectroscopic techniques. In pilot plant trials, 1-L quantities of whole milk were homogenized in a 2-stage homogenizer at 35°C (6.9 MPa/10.3 MPa) and, along with skim milk, were subjected to HTST pasteurization (72°C for 15 s) or UHT processing (135°C for 2 s). Other whole milk samples were processed using homogenization followed by either HTST pasteurization or UHT processing. The processed skim and whole milk samples were centrifuged further to remove fat and then acidified to pH 4.6 to isolate the corresponding whey fractions, and centrifuged again. The whey fractions were then purified using dialysis and investigated using the circular dichroism, Fourier transform infrared, and Trp intrinsic fluorescence spectroscopic techniques. Results demonstrated that homogenization combined with UHT processing of milk caused not only changes in protein composition but also significant secondary structural loss, particularly in the amounts of apparent antiparallel ß-sheet and α-helix, as well as diminished tertiary structural contact. In both cases of homogenization alone and followed by HTST treatments, neither caused appreciable chemical changes, nor remarkable secondary structural reduction. But disruption was evident in the tertiary structural environment of the whey proteins due to homogenization of whole milk as shown by both the near-UV circular dichroism and Trp intrinsic fluorescence. In-depth structural stability analyses revealed that even though processing of milk imposed little impairment on the secondary structural stability, the tertiary structural stability of whey protein was altered significantly. The following order was derived based on these studies: raw whole>HTST, homogenized, homogenized and pasteurized>skimmed and pasteurized, and skimmed UHT>homogenized UHT. The methodology demonstrated in this study can be used to gain insight into the behavior of milk proteins when processed and provides a new empirical and comparative approach for analyzing and assessing the effect of processing schemes on the nutrition and quality of milk and dairy product without the need for extended separation and purification, which can be both time-consuming and disruptive to protein structures.

Potential Probiotic Weizmannia coagulans WC10 Improved Antibiotic-Associated Diarrhea in Mice by Regulating the Gut Microbiota and Metabolic Homeostasis.

Antibiotic-associated diarrhea (AAD) is a common side effect of long-term and heavy antibiotic therapy. Weizmannia coagulans (W. coagulans) is an ideal probiotic because of its high viability, stability, and numerous health benefits to the host. In this study, the strains were first screened for W. coagulans WC10 (WC10) with a high combined ability based on their biological properties of gastrointestinal tolerance, adhesion, and short-chain fatty acid production ability. The effect of WC10 on mice with AAD was further evaluated. The results showed that WC10 was effective in improving the symptoms of AAD, effectively restoring antibiotic-induced weight loss, and reducing diarrhea status score and fecal water content. In addition, WC10 decreased the expression of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines, alleviated intestinal tissue damage and inflammation, and improved intestinal epithelial barrier function by decreasing serum levels of enterotoxin, DAO, and D-lactic acid, and by increasing the expression of the intestinal mucosal immune factors sIgA and occludin. Importantly, the composition and function of the gut microbiota gradually recovered after WC10 treatment, increasing the number of SCFAs-producing Bifidobacterium and Roseburia. Subsequently, the short-chain fatty acid (SCFA) content was examined and WC10 significantly increased acetate, propionate, and butyrate production. Additionally, metabolomic analysis also showed that WC10 reversed the antibiotic interference with major metabolic pathways. These findings provide a solid scientific basis for the future application of W. coagulans WC10 in the treatment of AAD.

Rapid UV Photo-Cross-Linking of α-Lactalbumin Hydrogel Biomaterial To Enable Wound Healing.

Although both the function and biocompatibility of protein-based biomaterials are better than those of synthetic materials, their usage as medical material is currently limited by their high costs, low yield, and low batch-to-batch reproducibility. In this article, we show how α-lactalbumin (α-LA), rich in tryptophan, was used to produce a novel type of naturally occurring, protein-based biomaterial suitable for wound dressing. To create a photo-cross-linkable polymer, α-LA was methacrylated at a 100-g batch scale with >95% conversion and 90% yield. α-LAMA was further processed using photo-cross-linking-based advanced processing techniques such as microfluidics and 3D printing to create injectable hydrogels, monodispersed microspheres, and patterned scaffolds. The obtained α-LAMA hydrogels show promising biocompatibility and degradability during in vivo testing. Additionally, the α-LAMA hydrogel can accelerate post-traumatic wound healing and promote new tissue regeneration. In conclusion, cheap and safe α-LAMA-based biomaterials could be produced, and they have a beneficial effect on wound healing. As a result, there may arise a potential partnership between the dairy industry and the development of pharmaceuticals.

Probiotic yogurt regulates gut microbiota homeostasis and alleviates hepatic steatosis and liver injury induced by high-fat diet in golden hamsters.

This study aimed to investigate the beneficial effects of probiotic yogurt on lipid metabolism and gut microbiota in metabolic-related fatty liver disease (MAFLD) golden hamsters fed on a high-fat diet (HFD). The results demonstrated that probiotic yogurt significantly reversed the adverse effects caused by HFD, such as body and liver weight gain, liver steatosis and damage, sterol deposition, and oxidative stress after 8 weeks of intervention. qRT-PCR analysis showed that golden hamsters fed HFD had upregulated genes related to adipogenesis, increased free fatty acid infiltration, and downregulated genes related to lipolysis and very low-density lipoprotein secretion. Probiotic yogurt supplements significantly inhibited HFD-induced changes in the expression of lipid metabolism-related genes. Furthermore, 16S rRNA gene sequencing of the intestinal content microbiota suggested that probiotic yogurt changed the diversity and composition of the gut microbiota in HFD-fed hamsters. Probiotic yogurt decreased the ratio of the phyla Firmicutes/Bacteroidetes, the relative abundance of the LPS-producing genus Desulfovibrio, and bacteria involved in lipid metabolism, whereas it increased the relative abundance of short-chain fatty acids producing bacteria in HFD-fed hamsters. Predictive functional analysis of the microbial community showed that probiotic yogurt-modified genes involved in LPS biosynthesis and lipid metabolism. In summary, these findings support the possibility that probiotic yogurt significantly improves HFD-induced metabolic disorders through modulating intestinal microflora and lipid metabolism and effectively regulating the occurrence and development of MAFLD. Therefore, probiotic yogurt supplementation may serve as an effective nutrition strategy for the treatment of patients with MAFLD clinically.

Copper-loaded Milk-Protein Derived Microgel Preserves Cardiac Metabolic Homeostasis After Myocardial Infarction.

Myocardial Infarction (MI) is a leading cause of death worldwide. Metabolic modulation is a promising therapeutic approach to prevent adverse remodeling after MI. However, whether material-derived cues can treat MI through metabolic regulation is mainly unexplored. Herein, a Cu2+ loaded casein microgel (CuCMG) aiming to rescue the pathological intramyocardial metabolism for MI amelioration is developed. Cu2+ is an important ion factor involved in metabolic pathways, and intracardiac copper drain is observed after MI. It is thus speculated that intramyocardial supplementation of Cu2+ can rescue myocardial metabolism. Casein, a milk-derived protein, is screened out as Cu2+ carrier through molecular-docking based on Cu2+ loading capacity and accessibility. CuCMGs notably attenuate MI-induced cardiac dysfunction and maladaptive remodeling, accompanied by increased angiogenesis. The results from unbiased transcriptome profiling and oxidative phosphorylation analyses support the hypothesis that CuCMG prominently rescued the metabolic homeostasis of myocardium after MI. These findings enhance the understanding of the design and application of metabolic-modulating biomaterials for ischemic cardiomyopathy therapy.

Supplementing maternal diet with milk oligosaccharides and probiotics helps develop the immune system and intestinal flora of offsprings.

Intestinal flora is very important for improving the development of the immune system in newborns. Maternal diet during pregnancy and lactation is one of the key factors affecting the growth and development of offspring. The objective of the present study was to examine whether supplementation of maternal diet with milk oligosaccharides and Bifidobacterium could influence the development of the intestinal flora and immune system of neonatal mice. In total, 30 pregnant Institute of Cancer Research (ICR) mice were randomly divided into six groups: a control group (basal diet) and five intervention groups (basal diet supplemented with different doses of 2'-fucosyllactose [2'-FL] and Bifidobacterium Bb12) during the pregnancy period. All female mice were monitored for physical health during gavage. After delivery, the number of mice in each litter, any deformity, and the development of the offspring were recorded. The spleen, blood, and fecal samples of six groups of 10-12 day-old offspring were collected. The results demonstrated that maternal milk oligosaccharides and probiotics conferred protective effects against lipopolysaccharide (LPS)-induced immunosuppression in mice offspring by significantly enhancing the immune organ indexes, splenocyte proliferation, immunoglobulin (immunoglobulin G, A, M) production as well as improving the macrophage phagocytosis (p < .05). The abundance of Lactobacilli and Bifidobacteria in the feces of offspring mice in the intervention groups was significantly higher than that of the offspring mice in the control group (p < .05). These findings suggest that the combination of 2'-FL and Bifidobacterium Bb12 displayed synergistic interactions between the two components that could promote the development of the immune system of the offsprings and improve their microbiota through maternal ingestion.

Biodegradation of conventional plastics: Candidate organisms and potential mechanisms.

With the benefits of coming at low-cost, being light-weight and having a high formability and durability, conventional plastics are widely used in both industry and daily life. However, because of their durability and extensive half-life with poor degradability and the low recycling rate, large amounts of plastic waste are accumulated in various environments, posing a significant threat to organisms and ecosystems. Compared to conventional physical and chemical degradation, biodegradation of plastic might become a promising and environmentally friendly solution for this problem. One of the aims of this review is to briefly describe the impact of plastics (especially microplastics). To facilitate rapid advancements in the area of plastic biodegradation, this paper provides a comprehensive review of the candidate organisms capable of biodegrading plastics and originating from four categories including natural microorganisms, artificially derived microorganisms, algae and animal organisms. In addition, the potential mechanism during plastic biodegradation and associated driving factors are summarized and discussed. Furthermore, the recent biotechnological progress (e.g. synthetic biology, systems biology, etc.) is highlighted as being key for future research. Finally, innovative research avenues for future studies are proposed. Concluding, our review is addressing the practical application of plastic biodegradation and the plastic pollution, thus necessitating more sustainable developments.

White-light crosslinkable milk protein bioadhesive with ultrafast gelation for first-aid wound treatment.

BACKGROUND: Post-traumatic massive hemorrhage demands immediately available first-aid supplies with reduced operation time and good surgical compliance. In-situ crosslinking gels that are flexibly adapting to the wound shape have a promising potential, but it is still hard to achieve fast gelation, on-demand adhesion, and wide feasibility at the same time. METHODS: A white-light crosslinkable natural milk-derived casein hydrogel bioadhesive is presented for the first time. Benefiting from abundant tyrosine residues, casein hydrogel bioadhesive was synthesized by forming di-tyrosine bonds under white light with a ruthenium-based catalyst. We firstly optimized the concentration of proteins and initiators to achieve faster gelation and higher mechanical strength. Then, we examined the degradation, cytotoxicity, tissue adhesion, hemostasis, and wound healing ability of the casein hydrogels to study their potential to be used as bioadhesives. RESULT: Rapid gelation of casein hydrogel is initiated with an outdoor flashlight, a cellphone flashlight, or an endoscopy lamp, which facilitates its usage during first-aid and minimally invasive operations. The rapid gelation enables 3D printing of the casein hydrogel and excellent hemostasis even during liver hemorrhage due to section injury. The covalent binding between casein and tissue enables robust adhesion which can withstand more than 180 mmHg blood pressure. Moreover, the casein-based hydrogel can facilitate post-traumatic wound healing caused by trauma due to its biocompatibility. CONCLUSION: Casein-based bioadhesives developed in this study pave a way for broad and practical application in emergency wound management.

Comparison of sensitization between beta-lactoglobulin and its hydrolysates.

BACKGROUND: Bovine beta-lactoglobulin is one of the first foreign antigens encountered by some newborn children, and it has been described as the main allergenic protein in cow's milk even when present at low concentration. OBJECTIVE: Enzymatic hydrolysis has been identified to be a very effective way to reduce the sensitization of beta-lg compared to other treatments. The aim of this study was to explore whether enzymatic hydrolysis could reduce the allergenicity of native bovine beta-lg. METHODS: beta-lg was hydrolyzed by trypsin. Twenty four BALB/c mice were divided into three groups and orally sensitized by native bovine beta-lg and its hydrolysates five times at weekly intervals. RESULTS: During the sensitization period, many serious systemic anaphylactic symptoms were observed in mice sensitized by native beta-lg compared to hydrolysates of beta-lg. Mice sensitized by hydrolysates of beta-lg showed a significantly lower spleen lymphocyte proliferation level than intact beta-lg. The beta-lg-specific IgE antibody levels in serum and intestinal fluid samples induced by native beta-lg were significantly elevated. Plasma histamine levels were also evaluated and showed the same trend as IgE. Moreover, the hydrolysates of beta-lg significantly up-regulated IFN-gamma and IL-10 production and down-regulated IL-4 and IL-5 secretions by murine splenocytes. CONCLUSION: These results suggested that enzymatic hydrolysis could partly reduce the allergenicity of beta-lg.

Prevalence and characterization of Salmonella from meat in slaughterhouses in Hangzhou, China.

Salmonella is an essential food-borne pathogenic microorganism. Humans could get infected by consuming of Salmonella-contaminated foods, especially contaminated meat. In this study, a total of 580 retail meat samples (280 pork, 240 chicken, and 60 goose) were collected from slaughterhouses in Hangzhou to determine the prevalence and antimicrobial susceptibility patterns of Salmonella. Isolates were characterized by serotyping, PFGE (Pulsed-field gel electrophoresis), Polymerase chain reaction (PCR), and antibiotic susceptibility testing. The rates of Salmonella-positive pork, chicken, and goose samples were 21.1% (n = 59), 10.4% (n = 25) and 10.0% (n = 6), respectively. The prevalence of Salmonella was 15.5% in slaughterhouses. Thirteen different Salmonella serovars were identified, and 6 isolates could not be identified. The most commonly prevalent serovars are Salmonella Rissen (S. Rissen) (n = 20, 22.2%), S. Derby (n = 16, 17.8%) and S. Typhimurium (n = 12, 13.3%). The detection rate of the remaining serovars ranged from 1.1%-11.11%. All Derby, Corvallis, and Kentucky strains were from pork. Seventy-two isolates (80.0%) showed drug resistance (DR) to at least one antibiotics, 19 (21.1%) were multi-drug resistant strains, 2 (2.2%) showed seven or more. The detection rate of Trimethoprim/Sulfamethoxazole-resistance (70.0%) was highest, followed by Ampicillin (55.6%). Salmonella resistance was found related to serovar and origin. The positive rates of DR gene sul1, sul2, sul3, class I integrons and blaTEM were 92.2%, 95.6%, 86.7%, 83.3% and 62.2%, respectively. Fifty-seven different PFGE patterns and 7 main clusters were obtained. This study revealed the high positive rates of Salmonella resistance and related DR genes, especially for Sulfamethoxazole-resistance and its related gene.

Lactobacillus plantarum ZJUIDS14 alleviates non-alcoholic fatty liver disease in mice in association with modulation in the gut microbiota.

This present study was designed to explore the protective role of Lactobacillus plantarum ZJUIDS14 against Non-alcoholic Fatty Liver Disease (NAFLD) in a high-fat-diet (HFD)-induced C57BL/6 mice model. The probiotic (109 CFU/every other day) was administered by oral gavage for 12 weeks. We found that L. plantarum ZJUIDS14 intervention significantly alleviated HFD related hepatic steatosis, liver damage, insulin resistance, and increased hepatic expression of peroxisome proliferator activated receptor α (PPAR-α) while stimulating the activation of AMP-activated protein kinase (AMPK). Furthermore, L. plantarum ZJUIDS14 improved mitochondrial function as reflected by an increase in dynamin related protein 1 (DRP1) and a decrease of proteins associated with oxidative phosphorylation (OXPHOS) after the treatment. Additionally, mice from the L. plantarum ZJUIDS14 group had a restored intestinal flora and homeostasis involving Coprostanoligenes group, Ruminococcaceae UCG-014, Allobaculum, Ruminiclostridium 1, and Roseburia. Meanwhile, these five genera exhibited a significant (negative or positive) association with ileum inflammation mRNA levels and SCFA contents, by Spearman's correlation analysis. In general, our data demonstrated that L. plantarum ZJUIDS14 mitigates hepatic steatosis and liver damage induced by HFD. Specifically, they strengthened the integrity of the intestinal barrier, regulated gut microbiota, and improved mitochondrial function. Our data provide an experimental basis for L. plantarum ZJUIDS14 as a promising candidate to prevent NAFLD.

Macrophage activation by exopolysaccharides from Streptococcus thermophilus fermented milk through TLRs-mediated NF-κB and MAPK pathways.

Lactic acid bacteria-derived exopolysaccharides are known for stimulating immune responses. In our previous study, a novel exopolysaccharide (EPS-3A) from skimmed milk fermented by the strain Streptococcus thermophilus (ZJUIDS-2-01) was extracted and structurally characterized. The present study aimed to investigate the effects of EPS-3A on macrophage activation and identify the underlying mechanism. EPS-3A was observed to promote TNF-α secretion and phagocytic uptake. RNA-seq analysis identified 949 differentially expressed genes (DEGs). GO analysis revealed that the DEGs were mainly enriched in metabolic pathways relating to the immune system. PPI network, KEGG pathway, western blot and functional verification assays indicated that MAPK and NF-κB were the key regulators modulating the expressions of the core gene TNF-α. Role and function of TLR2 and TLR4 for the recognition of EPS-3A were also determined. In conclusion, EPS-3A activated macrophages through MAPKs and NF-κB signaling mediated at least partly via TLR2 and TLR4.

Lactobacillus plantarum ZY08 relieves chronic alcohol-induced hepatic steatosis and liver injury in mice via restoring intestinal flora homeostasis.

This present study was designed to test the protective role of two Lactobacillus plantarum strains, E680 and ZY08, against alcoholic liver disease (ALD) in C57BL/6 mice. The ALD mouse model was established by exposing the mice to a Lieber-DeCarli ethanol liquid diet. The two probiotic strains (109 cfu/day) were administered by oral gavage, respectively. Our data showed that L. plantarum ZY08, but not E680, intervention significantly mitigated alcohol-related hepatic steatosis, liver injury, intestinal barrier, and it alleviated plasma endotoxin (LPS) levels, and affected hepatic genes relating to lipid metabolism. Furthermore, Lactobacillus plantarum ZY08 effectively restored intestinal flora homeostasis via recovering flora abundance, including Blautia, Oscillibacter, Lachnoclostridium and Intestimonas, and consequently elevated intestinalshort-chain fatty acid (SCFA) content. More importantly, removing intestinal microorganisms through ABX gavage markedly abolished the beneficial aspects of Lactobacillus plantarum ZY08, indicating that the regulative role of Lactobacillus plantarum ZY08 contributed to its protective role against ALD. Overall, Lactobacillus plantarum ZY08 is a potential candidate for mitigating alcohol-induced hepatic steatosis and liver injury.

Scalable Milk-Derived Whey Protein Hydrogel as an Implantable Biomaterial.

There are limited naturally derived protein biomaterials for the available medical implants. High cost, low yield, and batch-to-batch inconsistency, as well as intrinsically differing bioactivity in some of the proteins, make them less beneficial as common implant materials compared to their synthetic counterparts. Here, we present a milk-derived whey protein isolate (WPI) as a new kind of natural protein-based biomaterial for medical implants. The WPI was methacrylated at 100 g bench scale, >95% conversion, and 90% yield to generate a photo-cross-linkable material. WPI-MA was further processed into injectable hydrogels, monodispersed microspheres, and patterned scaffolds with photo-cross-linking-based advanced processing methods including microfluidics and 3D printing. In vivo evaluation of the WPI-MA hydrogels showed promising biocompatibility and degradability. Intramyocardial implantation of injectable WPI-MA hydrogels in a model of myocardial infarction attenuated the pathological changes in the left ventricle. Our results indicate a possible therapeutic value of WPI-based biomaterials and give rise to a potential collaboration between the dairy industry and the production of medical therapeutics.

Effect of Yellow Wine Lees Supplementation on Milk Antioxidant Capacity and Hematological Parameters in Lactating Cows under Heat Stress.

Fifteen multiparous lactating Chinese Holstein dairy cows were used in a replicated 3 × 3 Latin Square Design to evaluate the effect of total mixed rations (TMR) containing unfermented and fermented yellow wine lees (YWL) on the oxidative status of heat-stressed lactating cows and the oxidative stability of the milk and milk fatty acids they produced. Cows were fed with three isonitrogenous and isocaloric diets as follows: (1) TMR containing 18% soybean meal, (2) TMR containing 11% unfermented YWL (UM), and (3) TMR containing 11% fermented YWL (FM). The rectal temperature (at 1300 h) and respiratory rate were higher in control cows than in cows fed UM or FM. Both types of YWL were greater in total phenolic and flavonoid contents, reducing power, and radical scavenging abilities than soybean meal. Cows fed UM or FM had higher blood neutrophil, white blood cell, and lymphocyte counts, as well as lower plasma malondialdehyde level, higher plasma superoxide dismutase, glutathione peroxidase, and 2,2-diphenyl-1-picryl-hydrazyl-hydrate levels, and higher total antioxidant capacity in the plasma than those fed control diet. The proportion of milk unsaturated fatty acids was higher and that of saturated fatty acids was lower in UM- and FM-fed animals than in the control animals. Milk from UM- and FM-fed cows had lower malondialdehyde content but higher 2,2-diphenyl-1-picryl-hydrazyl-hydrate content than the control cows. In conclusion, feeding TMR containing UM and FM to cows reduced both the oxidative stress in heat-stressed cows and improved the oxidative capacity of their milk.