Effect of dietary heat-killed Lactiplantibacillus plantarum VSG3 on growth, immunity, antioxidant status, and immune-related gene expression in pathogen-aggravated Cyprinus carpio.

Heat-killed probiotics offer an alternative approach to enhance growth and disease resistance in farmed fish. In this study, we isolated Lactiplantibacillus plantarum VSG3 from the gut of Labeo rohita to investigate the effects of heat-killed L. plantarum (HK-LP) on the health and growth performance of Cyprinus carpio fingerlings. Different concentrations of HK-LP (0, 50, 100, 200, 300, and 400 mg/kg) were administered to the fish, followed by a challenge with Aeromonas hydrophila after 8 weeks of feeding. Notably, the LP200 group exhibited significantly improved percentage weight gain and specific growth rate, accompanied by the lowest feed conversion ratio. Post-challenge survival rates were considerably enhanced in the LP200 group, reaching 60.65%. Moreover, serum analysis indicated significantly higher levels of total protein and albumin in the LP200 group than in the control group. Although HK-LP had no substantial impact on certain serum parameters (glucose, total cholesterol, cortisol, and alanine aminotransferase), aspartate aminotransferase levels were considerably low in the LP200 group. Intestinal protease and trypsin activities significantly increased in the LP200 group, while no significant changes were observed in lipase and amylase activities post-pathogen challenge. Serum immunological indices, including lysozyme, alternative complement pathway, and phagocytic activity, improved considerably in the LP200 group. Additionally, serum antioxidant enzyme activities (superoxide dismutase [SOD], glutathione peroxidase [GPx], catalase [CAT], and myeloperoxidase) were significantly elevated in the LP200 group, while malondialdehyde level was reduced. Gene expression analysis in liver tissue indicated strong upregulation of antioxidant-related genes (SOD, CAT, nuclear factor erythroid 2 [NFE2]-related factor 2 [Nrf2], Kelch-like ECH-associated protein 1[Keap1]) in the LP100 and LP200 groups. Pro-inflammatory cytokines (IL-1ß and TNF-α) were considerably downregulated in the kidneys of the LP200 post-challenged fish, although the anti-inflammatory cytokine IL-10 showed an increased expression. Quadratic regression analysis identified the optimal dietary HK-LP level for maximizing growth and immune performance (200.381-270.003 mg/kg). In summary, our findings underscore the potential of HK-LP as a valuable dietary supplement for enhancing carp aquaculture, particularly at the appropriate concentration.

Production of prebiotic enriched maple syrup through enzymatic conversion of sucrose into fructo-oligosaccharides.

Maple syrup, a popular natural sweetener has a high content of sucrose, whose consumption is linked to different health issues such as obesity and diabetes. Hence, within this paper, the conversion of sucrose to prebiotics (fructo-oligosaccharides, FOS) was proposed as a promising approach to obtaining a healthier, value-added product. Enzymatic conversion was optimized with respect to key experimental factors, and thereafter derived immobilized preparation of fructosyltransferase (FTase) from Pectinex® Ultra SP-L (FTase-epoxy Purolite, 255 IU/g support) was successfully utilized to produce novel functional product in ten consecutive reaction cycles. The product, obtained under optimal conditions (60 °C, 7.65 IU/mL, 12 h), resulted in 56.0% FOS, 16.7% sucrose, and 27.3% monosaccharides of total carbohydrates, leading to a 1.6-fold reduction in caloric content. The obtained products` prebiotic potential toward the probiotic strain Lactobacillus plantarum 299v was demonstrated. The changes in physico-chemical and sensorial characteristics were esteemed as negligible.

Antimicrobial Cyclic Dipeptides from Japanese Quail (Coturnix japonica) Eggs Supplemented with Probiotic Lactobacillus plantarum.

Fifteen cyclic dipeptides (CDPs) containing proline, one cyclo(Phe-Ala) without proline, and a non-peptidyl DL-3-phenyllactic acid were previously identified in the culture filtrates of Lactobacillus plantarum LBP-K10, an isolate from kimchi. In this study, we used Japanese quail (Coturnix japonica) eggs to examine the effects of probiotic supplementation on the antimicrobial CDPs extracted from quail eggs (QE). Eggshell-free QE were obtained from two distinct groups of quails. The first group (K10N) comprised eggs from unsupplemented quails. The second group (K10S) comprised eggs from quails supplemented with Lb. plantarum LBP-K10. The QE samples were extracted using methylene chloride through a liquid-liquid extraction process. The resulting extract was fractionated into 16 parts using semi-preparative high-performance liquid chromatography. Two fractions, Q6 and Q9, were isolated from K10S and identified as cis-cyclo(L-Ser-L-Pro) and cis-cyclo(L-Leu-L-Pro). The Q9 fraction, containing cis-cyclo(L-Leu-L-Pro), has shown significant inhibitory properties against the proliferation of highly pathogenic multidrug-resistant bacteria, as well as human-specific and phytopathogenic fungi. Some of the ten combinations between the remaining fourteen unidentified fractions and two fractions, Q6 and Q9, containing cis-cyclo(L-Ser-L-Pro) and cis-cyclo(L-Leu-L-Pro) respectively, demonstrated a significant increase in activity against multidrug-resistant bacteria only when combined with Q9. The activity was 7.17 times higher compared to a single cis-cyclo(L-Leu-L-Pro). This study presents new findings on the efficacy of proline-containing CDPs in avian eggs. These CDPs provide antimicrobial properties when specific probiotics are supplemented.

Lactobacillus plantarum E2 regulates intestinal microbiota and alleviates Pseudomonas plecoglossicida induced inflammation and apoptosis in zebrafish (Danio rerio).

Pseudomonas plecoglossicida infection is a highly contagious epidemic in aquaculture, causing significant mortality among teleost. Our previous research has demonstrated that Lactobacillus plantarum E2 is beneficial for large yellow croaker in resisting infections caused by P. plecoglossicida. However, the relevant mechanisms remain largely unclear. In the present study, we used zebrafish (Danio rerio) to further explore the function of L. plantarum E2 and its mechanisms for resisting P. plecoglossicida infection. E2 supplementation diet significantly improved the growth rates and α-amylase and trypsin activities of the liver in zebrafish. After challenge with P. plecoglossicida strain PQLYC4, the survival rates of zebrafish were improved, and immune-related genes expression (IL-1ß, TNF-α, IL-8, Ig-Z, TLR-22 and IL-12α) were down-regulated. Histological analysis showed that E2 group had a longer intestinal villus and thicker intestinal walls after 30 days of feeding and healthier intestinal structure after challenge with P. plecoglossicida strain PQLYC4. Furthermore, co-incubation of zebrafish embryo fibroblast (ZF-4 cells) with L. plantarum E2 reduced apoptosis of ZF-4 cells after exposed to P. plecoglossicida. Intestinal microbiota analysis showed that E2 strain significantly increased the relative abundance of Lactobacillus and Pseudomonas, and PCoA analysis revealed a noticeable divergence in the intestinal microbial communities after E2 supplement. Together, our results suggested that E2 strain may promote zebrafish survival against P. plecoglossicida infection by regulating the intestinal microbiota and alleviating inflammatory response and apoptosis, thus exhibiting the potential as a probiotic.

Purification and characterization of a novel low-molecular-weight antimicrobial peptide produced by Lactiplantibacillus plantarum NMGL2.

The present study aimed to purify and characterize a novel low-molecular-weight antimicrobial peptide (AMP) named as PNMGL2 produced by Lactiplantibacillus plantarum NMGL2. The AMP was effectively separated and purified by ethyl acetate extraction and DEAE-Sepharose anion exchange chromatography. Tricine-SDS-PAGE of the purified AMP showed a major protein band below 1.7 kDa, which was identified by MALDI-TOF MS to be a hexapeptide LNFLKK (761.95 Da), and structurally characterized to be combination of helixes and random coil by a PEP-FOLD 3 De novo approach. The antimicrobial activity of LNFLKK was confirmed by chemical synthesis of the peptide that showed clear inhibition (MIC 7.8 mg/mL) against both Gram-positive bacteria (Staphylococcus aureus and Listeria monocytogenes), and Gram-negative bacteria (Enterobacter sakazakii, Escherichia coli and Shigella flexneri). PNMGL2 was pH resistant (pH 2-9), heat stable (121 °C, 30 min), and protease sensitive. Treatment of UV rays, sodium chloride and organic solvents did not decrease the activity. Sequencing of the whole genome of L. plantarum NMGL2 revealed presence of a bacteriocin gene cluster with two putative bacteriocin genes (ORF4 and ORF5) that were not expressed, confirming the significance of PNMGL2 contributing the antimicrobial activity of the strain. This study demonstrated the low-molecular-weight AMP that was uncharacterized in the relevant available databases, suggesting its potential application as a novel natural food preservative.

Structural characteristics and biological activity of lactic acid bacteria exopolysaccharides separated by ethanol/(NH4)2SO4 ATPS.

Exopolysaccharides (EPS) from lactic acid bacteria (LAB) as edible and safe bioproducts with health benefits have become an interesting topic. In this study, aqueous two-phase system (ATPS) was established using ethanol and (NH4)2SO4 as phase-forming substances to separate and purify LAB EPS from Lactobacillus plantarum 1.0665. The operating conditions were optimized by a single factor and response surface method (RSM). The results indicated that an effectively selective separation of LAB EPS was achieved by the ATPS consisted of 28 % (w/w) ethanol and 18 % (w/w) (NH4)2SO4 at pH 4.0. Under optimized conditions, the partition coefficient (K) and recovery rate (Y) were well matched with the predicted value of 3.83 ± 0.019 and 74.66 ± 1.05 %. The physicochemical properties of purified LAB EPS were characterized by various technologies. According to the results, LAB EPS was a complex polysaccharide with a triple helix structure mainly composed of mannose, glucose and galactose in the molar ratio of 1.00: 0.32: 0.14, and it proved that the ethanol/(NH4)2SO4 system had good selectivity for LAB EPS. In addition, LAB EPS displayed excellent antioxidant activity, antihypertension activity, anti-gout capacity and hypoglycemic activity in vitro analysis. The results suggested that LAB EPS could be a dietary supplement applied in functional foods.

Embryonic injection of Lactobacillus plantarum PA01 alters the microbial diversity in the gastrointestinal tract of the broilers before and after hatching.

The total number of intestinal microbiotas is low, and the intestinal tract develops rapidly and imperfectly at the embryonic stage. Embryonic period as a particular physiological stage is an important time window to explore how to regulate organismal health by probiotics. Therefore, this experiment was conducted to investigate the effect of embryonic injection of Lactobacillus plantarum PA01 at embryonic d 14 (E14) on the microbiome of the contents of the gizzard, cecum at embryonic d 20 (E20) and cecum at d 1 posthatch (D1) by 16S rRNA sequencing. Results showed that PA01 had no significant effect on broiler body weight and yolk sac weight at E20 and D1 (P > 0.05). PA-01 altered the Shannon index and ß diversity of the gizzard at E20 (P < 0.05), increased the abundance of Firmicutes (P < 0.05), and decreased the relative abundance of Proteobacteria, Bacteroidota, and Actinobacteriota (P < 0.05). At the genus level of the microbiota, PA01 significantly increased the relative abundance of Lactiplantibacillus (P < 0.05). At 20 embryos, PA01 altered the α and ß diversity indices (P < 0.05) and decreased the relative abundance of Salmonella (P < 0.05) of the cecal microbiota. The biomarkers of PA01 group were Lactobacillales, Blautia, Lachnospiraceae, and Asinibacterium. Embryonic injection of PA01 altered the E20 intestinal microbes. PA01 altered the ß-diversity index of the 1-day-old cecum (P < 0.05), and there was no significant effect on microbial composition at the phylum and genus level (P > 0.05). LefSe analysis revealed that the biomarkers of the PA01 group were Lactobacillaceae, Lactiplantibacillus, Moraxellaceae, and Acinetobacter. Biomarkers in the Con group were Devosia, Bacillus, Nordella, Mesorhizobium, and Pseudolabrys. PA01 increased acetic acid in the gastrointestinal tract at E20 along with acetic and butyric acid in cecum of 1-day-old. In conclusion, embryo-injected L. plantarum PA01 altered the structure and metabolites of the microbial flora before and after hatching, in particular promoting the colonization of Lactobacillus.

High-speed electrospinning of phycocyanin and probiotics complex nanofibrous with higher probiotic activity and antioxidation.

This study reports for the first time the co-encapsulation of probiotics and phycocyanin by electrospinning. SEM showed that the electrospun fibers exhibited a homogeneous, smooth surface and a circular shape. XRD and ATR-FTIR results showed that Lactiplantibacillus plantarum 1-24-LJ and Pc were co-embedded in the fibers and that the presence of L. plantarum 1-24-LJ promoted the encapsulation of phycocyanin. TG analysis showed that the addition of phycocyanin and L. plantarum 1-24-LJ improved the composite fiber's thermal stability. The fibers co-embedded with phycocyanin and L. plantarum 1-24-LJ had the highest DPPH and ABTS+ activity, indicating that the two may have synergistic antioxidant effects. After 28 days, the viability of the strain could still be above 6 log cfu/g, and the addition of phycocyanin could help to improve the strain's survivability. In this experiment, a co-embedding method for probiotics and antioxidants was proposed, which could effectively increase the survivability of probiotics and improve the antioxidant properties of the fibers.

Effects of Lactobacillus plantarum and its fermentation products on growth performance, immune function, intestinal pH, and cecal microorganisms of Lingnan yellow chicken.

The present research was conducted to investigate the effects of dietary supplementation of Lactobacillus plantarum and its fermentation products on growth performance, specific immune function, intestinal pH, and cecal microorganisms in yellow-feather broilers. A total of 1,200 yellow-feather broilers of similar weight and good health condition at 1 d of age were selected and randomly divided into 5 groups. The CK group was fed the basal diet, and the experimental group (I, II, III, IV) were supplemented with 0.1, 0.15% L. plantarum and 3, 4% L. plantarum fermentation products. The results showed that each treatment could improve the growth performance (P < 0.05) and feed conversion rate of yellow-feather broilers. Besides, the pH value of the gastrointestinal tract of yellow-feather broilers (P < 0.05) was significantly reduced through the use of L. plantarum and its fermentation products as additives, which also facilitated the animals to regulate the balance of cecal microorganisms. The immune function assay showed that the bursal index (P < 0.05), spleen index (P < 0.05), and the content of serum immunoglobulins IgA and IgG (P < 0.05) were significantly increased in yellow-finned broilers aged 1 to 21 d by supplementing the diet with L. plantarum. In conclusion, adding L. plantarum or its fermentation products to the diet can improve the growth performance of yellow-feather broilers, and the direct addition of L. plantarum is better than adding fermentation products.

Lactobacillus plantarum HF02 alleviates lipid accumulation and intestinal microbiota dysbiosis in high-fat diet-induced obese mice.

BACKGROUND: Obesity is closely associated with lipid accumulation and intestinal microbiota dysbiosis. It has been proved that probiotics supplement contributes to alleviate obesity. The objective of this study was to investigate the mechanism by which Lactobacillus plantarum HF02 (LP-HF02) alleviated lipid accumulation and intestinal microbiota dysbiosis in high-fat diet-induced obese mice. RESULTS: Our results showed that LP-HF02 ameliorated body weight, dyslipidemia, liver lipid accumulation, and liver injury in obese mice. As expected, LP-HF02 inhibited pancreatic lipase activity in small intestinal contents and increased fecal triglyceride levels, thereby reducing dietary fat hydrolysis and absorption. Moreover, LP-HF02 ameliorated the intestinal microbiota composition, as evidenced by the enhanced ratio of Bacteroides to Firmicutes, the decreased abundance of pathogenic bacteria (including Bacteroides, Alistipes, Blautia, and Colidextribacter) and the increased abundance of beneficial bacteria (including Muribaculaceae, Akkermansia, Faecalibaculum, and Rikenellaceae_RC9_gut_group). LP-HF02 also increased fecal short-chain fatty acids (SCFAs) levels and colonic mucosal thickness, and subsequently decreased serum lipopolysaccharide (LPS), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α) levels in obese mice. Additionally, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot results demonstrated that LP-HF02 ameliorated hepatic lipid accumulation via activating the adenosine monophosphate (AMP)-activated protein kinase (AMPK) pathway. CONCLUSION: Therefore, our results indicated that LP-HF02 could be considered as a probiotic preparation for preventing obesity. © 2023 Society of Chemical Industry.

Optimization, Probiotic Characteristics, and Rheological Properties of Exopolysaccharides from Lactiplantibacillus plantarum MC5.

This study optimized the exopolysaccharides (EPS) production for Lactiplantibacillus plantarum MC5 (Lp. plantarum MC5) and evaluated the resistance to human simulated digestive juices, antioxidant activity in vitro, and rheological properties of EPS-MC5. The results showed that maximum EPS production of 345.98 mg/L (about 1.5-old greater than the initial production) was obtained at optimal conditions of inoculum size (4.0%), incubation time (30 h), incubation temperature (34.0 °C), and initial pH value (6.40). Furthermore, the resisting-digestion capacity of EPS-MC5 after 180 min in α-amylase, simulated gastric juice (pH 2.0, 3.0, 4.0), and simulated intestinal juice (pH 6.8) was 98.59%, 98.62%, 98.78%, 98.86%, and 98.74%, respectively. In addition, the radical scavenging rates of DPPH•, ABTS•, •OH, and ferric-iron reducing power (OD700) of EPS-MC5 were 73.33%, 87.74%, 46.07%, and 1.20, respectively. Furthermore, rheological results showed that the EPS-MC5 had a higher apparent viscosity (3.01 Pa) and shear stress (41.78 Pa), and the viscoelastic modulus (84.02 and 161.02 Pa at the shear frequency of 100 Hz). These results provide a new insight into the application of EPS in human health and functional foods, which could also improve theoretical guidance for the industrial application of EPS.

Optimization of bacteriocin production by Lactobacillus plantarum using Response Surface Methodology.

Bacteriocin production is influenced by various factors such as carbon and nitrogen sources as well as fermentation conditions including pH, temperature, and agitation-these factors aid in optimizing bacteriocin production and improving its inhibitory activity against pathogens for great economic significance. The study investigates the effect of growth conditions on bacteriocin production by Lactobacillus plantarum. The response surface methodology was applied to optimize and determine the interaction among the process variables in bacteriocin production by Lactobacillus plantarum and determine their optimum levels. Chloroform-Methanol (2:1 v/v) was used for crude bacteriocin extraction through the liquid-liquid extraction method, and its antimicrobial activity was assessed. The sample has shown inhibitory activity against all the organism,s i.e., E. coli, S. aureus, K. pneumoniae and P. aeruginosa good diffusion. Compared to control and ciprofloxacin, the sample at all concentrations (Lab 9, 1%, 2%, 3%) has shown better inhibitory activity in pH-7 and-8 at 37 °C against S.aureus in good diffusion. The selected factors were carbon and nitrogen source, temperature, and pH. The bacteriocin was produced at maximum activity in MRS broth supplemented with 1% dextrose and 1% ammonium nitrate. RSM analysis found that the optimal temperature for bacteriocin production was 36°C at a pH of 6.5 using 1% inoculum. At the same time, the increase in the percentage of inoculum (2% and 3%) did not affect bacteriocin production. The quadratic model found that temperature and pH profoundly affected bacteriocin production.

Preparation of alginate-whey protein isolate and alginate-pectin-whey protein isolate composites for protection and delivery of Lactobacillus plantarum.

Probiotics are sensitive to external conditions, resulting in low survival rates after being ingested or during food production, transportation and storage. In order to improve the survival rate of Lactobacillus plantarum (LP) during gastrointestinal digestion, storage, and freeze-drying, alginate-whey protein isolate (ALG-WPI) and alginate-pectin-whey protein isolate (ALG-PEC-WPI) composites were employed to encapsulate LP. The encapsulation efficiency of ALG-WPI-LP and ALG-PEC-WPI-LP beads both reached more than 99 %. Scanning electron microscopy (SEM) indicated that dense and rough aggregates were formed on the surface of both composites, and attached LP cells could be observed inside the beads. The ALG-WPI and ALG-PEC-WPI composites can protect the viability of LP in simulated gastric fluid (SGF) and release the probiotics in simulated intestinal fluid (SIF). The storage stability of LP at 4 °C was improved by about 15 % in comparison with bare LP and the survival rates of LP in ALG-WPI-LP and ALG-PEC-WPI-LP powders after freeze-drying were increased by 65.37 % and 72.06 %, respectively. The formation mechanism of ALG-WPI and ALG-PEC-WPI composites was further explored by fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and thermogravimetric analysis (TGA). The ALG-WPI and ALG-PEC-WPI composites have great potential to protect and deliver probiotics in food systems.

Purification, amino acid sequence, and characterization of bacteriocin GA15, a novel class IIa bacteriocin secreted by Lactiplantibacillus plantarum GCNRC_GA15.

The bacteriocins produced by lactic acid bacteria (LAB) are attracting attention due to their promising applications in food and pharmaceuticals fields. Hence, a LAB strain, GCNRC_GA15, was isolated from Egyptian goat cheese, and molecularly identified as Lactiplantibacillus plantarum. This strain showed a wide antimicrobial spectrum, which was found to be of proteineous nature, suggesting that L. plantarum GCNRC_GA15 is a bacteriocin-producer. This bacteriocin (bacteriocin GA15) was partially purified using cation exchange, and hydrophobic interaction chromatography. Tricine SDS-PAGE analysis for the fraction showing bacteriocin activity has estimated the molecular mass to be 4369 Da. Furthermore, amino acid sequencing of this peptide has detected 34 amino acids, and comparing its amino acid sequence with those of some pediocin-like bacteriocins revealed that bacteriocin GA15 has the conserved sequence (YYGNGV/L) in its N-terminal region which identified bacteriocin GA15 as a pediocin-like bacteriocin. Bacteriocin GA15 showed good heat and pH stabilities, and its activity was enhanced after treatment with Tween 80 or Triton X-100. Bacteriocin production medium was statistically optimized using the Plackett-Burman and Central Composite designs. As a result, bacteriocin production increased from 800 to 12,800 AU/ml using the optimized medium in comparison with result recorded for the un-optimized medium.

Purification and characterization of Lactobacillus plantarum-derived bacteriocin with activity against Staphylococcus argenteus planktonic cells and biofilm.

Bacteriocins inhibit various foodborne bacteria in planktonic and biofilm forms. However, bacteriocins with antibacterial and antibiofilm activity against Staphylococcus argenteus, a pathogen that can cause food poisoning, are still poorly known. Here, the novel bacteriocin LSB1 derived from Lactobacillus plantarum CGMCC 1.12934 was purified and characterized extensively. LSB1 had a molecular weight of 1425.78 Da and an amino acid sequence of YIFVTGGVVSSLGK. Moreover, LSB1 exhibited excellent stability under heat and acid-base stress and presented sensitivity to pepsin and proteinase K. LSB1 exhibited an extensive antimicrobial spectrum against both Gram-positive and Gram-negative bacteria. Minimum inhibitory concentration of LSB1 against S. argenteus_70917 was 10.36 µg/ml, which was lower than that of most of the previously found bacteriocins against Staphylococcus strains. Furthermore, LSB1 significantly inhibited S. argenteus_70917 planktonic cells (p < 0.01) and decreased their viability. Scanning electron microscopy analysis revealed that cell membrane permeability of S. argenteus_70917 upon exposure to LSB1 showed leakage of cytoplasmic contents and rupture, leading to cell death. In addition, biofilm formation ability of S. argenteus_70917 was significantly (p < 0.01) impaired by LSB1, with the percent inhibition of 35% at 10 µg/ml and 80% at 20 µg/ml. Overall, this study indicates that LSB1 can be considered a potential antibacterial agent in the control of S. argenteus in both planktonic and biofilm states. PRACTICAL APPLICATION: Foodborne pathogenic bacteria, such as Staphylococcus argenteus, and their biofilms represent potential risks for food safety. In recent years, customers' demand for "natural" products has increased food control. This study describes the novel bacteriocin LSB1 produced by the lactic acid bacterium species Lactobacillus plantarum. LSB1 showed strong antibacterial and antibiofilm activity against S. argenteus as well as thermal and acid-alkaline stability. Furthermore, the mechanisms of action of LSB1 on S. argenteus were preliminarily explored. These results indicate that LSB1 might be potentially used as an effective and natural food preservative.

Isolation and identification of new source of bacteriocin-producing Lactobacillus plantarum C010 and growth kinetics of its batch fermentation.

The control of food-borne pathogens and spoilage organisms in meat and related products is urgently needed. Bacteriocins produced by lactic acid bacteria (LAB) are promising natural food preservatives. In this study, six bacteriocin-producing bacteria were screened from soil and fresh cow dung. Pseudomonas koreensis PS1, a specific spoilage organism from spoiled chilled pork, was used as the indicator bacteria. From the analyses, the strain C010 was selected due to its high yield, broad spectrum, and subculture stability. Through morphological, biochemical, and 16S rDNA gene sequence analysis, this strain was identified as Lactobacillus plantarum. Crude bacteriocin extracted from the cell-free supernatant (CFS) of L. plantarum C010 was stable under high temperature, ultraviolet radiation, and protease attack (pepsin, trypsin, and proteinase K). The kinetics of bacterial growth and bacteriocin production by L. plantarum C010 were analyzed during batch fermentation. Bacteriocin was produced throughout the logarithmic growth phase, and the Leudeking-Piret model characterized the synthesis of bacteriocins. The present study indicates that this novel bacteriocin produced by bacteria is a promising option for reducing spoilage microorganisms and can be widely used as a bio-preservative in meat and other foods.

A Potential Synbiotic Strategy for the Prevention of Type 2 Diabetes: Lactobacillus paracasei JY062 and Exopolysaccharide Isolated from Lactobacillus plantarum JY039.

The disturbance of intestinal microorganisms and the exacerbation of type 2 diabetes (T2D) are mutually influenced. In this study, the effect of exopolysaccharides (EPS) from Lactobacillus plantarum JY039 on the adhesion of Lactobacillus paracasei JY062 was investigated, as well as their preventive efficacy against T2D. The results showed that the EPS isolated from L. plantarum JY039 effectively improved the adhesion rate of L. paracasei JY062 to Caco-2 cells (1.8 times) and promoted the proliferation of L. paracasei JY062. In the mice experiment, EPS, L. paracasei JY062 and their complex altered the structure of the intestinal microbiota, which elevated the proportion of Bifidobacterium, Faecalibaculum, while inversely decreasing the proportion of Firmicutes, Muribaculaceae, Lachnospiraceae and other bacteria involved in energy metabolism (p < 0.01; p < 0.05); enhanced the intestinal barrier function; promoted secretion of the gut hormone peptide YY (PYY) and glucagon-like peptide-1 (GLP-1); and reduced inflammation by balancing pro-inflammatory factors IL-6, TNF-α and anti-inflammatory factor IL-10 (p < 0.01; p < 0.05). These results illustrate that EPS and L. paracasei JY062 have the synbiotic potential to prevent and alleviate T2D.

Differential Effects of Transition Metals on Growth and Metal Uptake for Two Distinct Lactobacillus Species.

Lactobacillus is a genus of Gram-positive bacteria and comprises a major part of the lactic acid bacteria group that converts sugars to lactic acid. Lactobacillus species found in the gut microbiota are considered beneficial to human health and commonly used in probiotic formulations, but their molecular functions remain poorly defined. Microbes require metal ions for growth and function and must acquire them from the surrounding environment. Therefore, lactobacilli need to compete with other gut microbes for these nutrients, although their metal requirements are not well-understood. Indeed, the abundance of lactobacilli in the microbiota is frequently affected by dietary intake of essential metals like zinc, manganese, and iron, but few studies have investigated the role of metals, especially zinc, in the physiology and metabolism of Lactobacillus species. Here, we investigated metal uptake by quantifying total cellular metal contents and compared how transition metals affect the growth of two distinct Lactobacillus species, Lactobacillus plantarum ATCC 14917 and Lactobacillus acidophilus ATCC 4356. When grown in rich or metal-limited medium, both species took up more manganese, zinc, and iron compared with other transition metals measured. Distinct zinc-, manganese- and iron-dependent patterns were observed in the growth kinetics for these species and while certain levels of each metal promoted the growth kinetics of both Lactobacillus species, the effects depend significantly on the culture medium and growth conditions. IMPORTANCE The gastrointestinal tract contains trillions of microorganisms, which are central to human health. Lactobacilli are considered beneficial microbiota members and are often used in probiotics, but their molecular functions, and especially those which are metal-dependent, remain poorly defined. Abundance of lactobacilli in the microbiota is frequently affected by dietary intake of essential metals like manganese, zinc, and iron, but results are complex, sometimes contradictory, and poorly predictable. There is a significant need to understand how host diet and metabolism will affect the microbiota, given that changes in microbiota composition are linked with disease and infection. The significance of our research is in gaining insight to how metals distinctly affect individual Lactobacillus species, which could lead to novel therapeutics and improved medical treatment. Growth kinetics and quantification of metal contents highlights how distinct species can respond differently to varied metal availability and provide a foundation for future molecular and mechanistic studies.

Potential application of lactic acid bacteria in the biopreservation of red grape from mycotoxigenic fungi.

BACKGROUND: Filamentous fungi are the main contamination agent in the viticultural sector. Use of synthetic fungicides is the regular answer to these contaminations. Nevertheless, because of several problems associated with the use of synthetic compounds, the industry demands new and safer methods. In the present work, the biopreservation potential of four lactic acid bacteria (LAB) strains was studied against the principal grape contaminant fungi. RESULTS: Agar diffusion test evidenced that all four culture-free supernatant (CFS) had antifungal properties against all tested fungi. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) test values evidenced that media fermented by the Lactobacillus plantarum E3 and Lactobacillus plantarum E4 strains showed the highest antifungal activity, resulting in an MFC from 6.3 to 100 g L-1 . Analysis of CFS evidenced the presence of different antifungal compounds, such as lactic acid, phenyllactic acid and pyrazines. In tests on red grapes, an average reduction of 1.32 log10 of the spores per gram of fruit was achieved by all CFS in grapes inoculated with Aspergillus ochraceus and by 0.94 log10 for L. plantarum E3 CFS against Botrytis cinerea. CONCLUSION: The antifungal activity of the fermented CFS by L. plantarum E3 reduced the growth of B. cinerea and A. ochraceus in grapes, which are the main contaminant and main producer of ochratoxin A in these crops, respectively. Therefore, based on the results obtained in this work, use of the strain L. plantarum E3 could be an interesting option for the biopreservation of grapes. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A four-probiotic preparation for ventilator-associated pneumonia in multi-trauma patients: results of a randomized clinical trial.

The role of probiotics in the prevention of ventilator-associated pneumonia (VAP) remains inconclusive. The aim of this study was to assess the efficacy of a probiotic regimen for VAP prophylaxis in mechanically ventilated multi-trauma patients, intubated immediately after the injurious insult. In a randomized, placebo-controlled study enrolling multi-trauma patients, patients expected to require mechanical ventilation for >10 days were assigned at random to receive prophylaxis with a probiotic formula (n=59) or placebo (n=53). The probiotic formula was a preparation of Lactobacillus acidophilus LA-5 [1.75 × 109 colony-forming units (cfu)], Lactobacillus plantarum (0.5 × 109 cfu), Bifidobacterium lactis BB-12 (1.75 × 109 cfu) and Saccharomyces boulardii (1.5 × 109 cfu) in sachets. Each patient received two sachets twice daily for 15 days: one through the nasogastric tube and one spread on the oropharynx. The incidence of VAP was the primary endpoint. The incidence of other infections and sepsis, and the duration of hospital stay were the secondary endpoints. Administration of probiotics reduced the incidence of VAP [11.9% vs 28.3%, hazard ratio (HR) 0.34, 95% confidence interval (CI) 0.13-0.92; P=0.034] and sepsis [6.8% vs 24.5%, odds ratio 0.22, 95% CI 0.07-0.74: P=0.016]. Furthermore, probiotic prophylaxis reduced the time of stay in the intensive care unit (ICU) and the length of hospital stay. The prophylactic use of probiotics with a combination of enteral and topical application to the oropharynx had a positive effect on the incidence of VAP and sepsis, as well as on ICU and total hospital stay in patients receiving protracted mechanical ventilation.