Probiotic Lactic Acid Bacteria Strains Research Articles

A comparative study on heavy metal (cadmium) tolerance and biosorption capacity of locally isolated lactic acid bacteria

Heavy metals (HM) have less biological role but show extreme toxicity in living systems. Lactic acid bacteria (LAB) are beneficial microorganisms, some of which have the ability to chelate HM and may be used to treat heavy metals poisoning. The aim of the current study was to select and identify Cd-removing probiotic LAB strains, for future pharmaceutical application. Seventeen LAB strains, from human excretions and animal fecal matter, were initially verified for their probiotic properties. The strains which could survive in the presence of Cd (≥100 ppm) were selected. The metal removal efficiency of isolates was compared, through atomic absorption spectrophotometry (AAS), by measuring the reduction in Cd contents of CdCl2 5 mg/L (3.05 ppm of Cd) supplemented broth following 24 h culturing of the strains. The Cd reduction of 38–75% was recorded in culture broth. The Cd removal ability significantly varied (p < 0.05) with microbial surface charge, incubation period, temperature and pH of media. Levilactobacillus brevis MW362790 and Pediococcus pentosaceus MT323062, identified through 16S rRNA gene sequencing, were recognized as strong Cd chelators. These strains may have application in pharmaceutical or agriculture (farm animals) for protection against cadmium toxicity. Further studies are suggested on in vivo metal detoxification potential.

Pistacia terebinthus MEYVESİ: GIDA BOZULMASINI ÖNLEMEK İÇİN BİR ALTERNATİF

The aim of this work was to investigate the usage possibilities of P. terebinthus fruit, which has a limited usage area, in the food industry. Antimicrobial activity of hexane extract obtained from P. terebinthus fruits from Adıyaman (Turkey) was determined by disc diffusion and micro-dilution assays against test microorganisms. In addition, the antibacterial activity of the extract on Escherichia coli O157:H7, one of the most important food-borne pathogens, was determined by viable cell count using a macro-dilution assay. The potential for use of the extract with probiotic candidate lactic acid bacteria (LAB) strains was also investigated. The hexane extract presented antimicrobial activity against all the tested microorganisms, with good inhibition zone diameters between 10.51 mm and 18.02 mm. MIC and MFC or MBC values of P. terebinthus fruit extract were determined as 5-80 µg/µL against all tested microorganisms. The lowest MIC and MBC values (5 µg/µL) of the extract were obtained against E. coli O157:H7. Macro-dilution assay results indicated that the P. terebinthus extract at various concentrations (5-10-20 mg/mL) inhibited the growth of E. coli O157:H7 more than the control group after all of the incubation hours. No viable E. coli O157:H7 cells were detected after 48 hours at all concentrations. The extract showed low antimicrobial activity, and relatively high bactericidal concentration on probiotic candidate LAB strains. This shows that P. terebinthus hexane extract at appropriate concentrations can be used together with probiotic strains as a natural preservative and biopreservative to prevent food spoilage and extend shelf life.

Species Identification of Potential Probiotic Lactic Acid Bacteria Isolated from Malaysian Fermented Food Based on 16S Ribosomal RNA (16S rRNA) and Internal Transcribed Spacer (ITS) Sequences

Taxonomic identification of potential probiotic lactic acid bacteria (LAB) in fermented foods is essential as the microorganisms’ benefits are varied and often strain-specific. Using biochemical and physiological methods alone is inadequate to precisely distinguish each strain. In this study, molecular techniques were employed in the identification of 17 LAB isolated from three Malaysian fermented foods: belacan (BE), bosou (BO), and budu (BUM). The respective sizes of polymerase chain reaction (PCR) products from the isolates were approximately 1500 bp and 750 bp when amplified with the 16S ribosomal RNA (16S rRNA) and internal transcribed spacer (ITS) gene primers. The phylogenetic analysis using both gene sequences revealed that all BE and BO isolates were identified as Lactiplantibacillus plantarum, while all BUM isolates were identified as Lacticaseibacillus paracasei. Both 16S rRNA and ITS genes could disclose the identity of the isolates up to the species level. In summary, the use of the ITS gene in conjunction with the 16S rRNA gene can help with the more effective identification of potential probiotic LAB strains isolated from fermented food.

Dual Role of Probiotic Lactic Acid Bacteria Cultures for Fermentation and Control Pathogenic Bacteria in Fruit-Enriched Fermented Milk.

The food industry has been developing new products with health benefits, extended shelf life, and without chemical preservation. Bacteriocin-producing lactic acid bacteria (LAB) strains have been evaluated for food fermentation to prevent contamination and increase shelf life. In this study, potentially probiotic LAB strains, Lactiplantibacillus (Lb.) plantarum ST8Sh, Lacticaseibacillus (Lb.) casei SJRP38, and commercial starter Streptococcus (St.) thermophilus ST080, were evaluated for their production of antimicrobial compounds, lactic acid and enzyme production, carbohydrate assimilation, and susceptibility to antibiotics. The characterization of antimicrobial compounds, the proteolytic activity, and its inhibitory property against Listeria (List.) monocytogenes and Staphylococcus (Staph.) spp. was evaluated in buriti and passion fruit-supplemented fermented milk formulations (FMF) produced with LAB strains. Lb. plantarum ST8Sh was found to inhibit List. monocytogenes through bacteriocin production and produced both L(+) and D(-) lactic acid isomers, while Lb. casei SJRP38 mainly produced L(+) lactic acid. The carbohydrate assimilation profiles were compatible with those usually found in LAB. The potentially probiotic strains were susceptible to streptomycin and tobramycin, while Lb. plantarum ST8Sh was also susceptible to ciprofloxacin. All FMF produced high amounts of L(+) lactic acid and the viability of total lactobacilli remained higher than 8.5 log CFU/mL during monitored storage period. Staph. aureus ATCC 43300 in fermented milk with passion fruit pulp (FMFP) and fermented milk with buritipulp (FMB), and Staph. epidermidis KACC 13234 in all formulations were completely inhibited after 14days of storage. The combination of Lb. plantarum ST8Sh and Lb. casei SJRP38 and fruit pulps can provide increased safety and shelf-life for fermented products, and natural food preservation meets the trends of the food market.

Recent advances in genetic tools for engineering probiotic lactic acid bacteria.

Synthetic biology has grown exponentially in the last few years, with a variety of biological applications. One of the emerging applications of synthetic biology is to exploit the link between microorganisms, biologics, and human health. To exploit this link, it is critical to select effective synthetic biology tools for use in appropriate microorganisms that would address unmet needs in human health through the development of new game-changing applications and by complementing existing technological capabilities. Lactic acid bacteria (LAB) are considered appropriate chassis organisms that can be genetically engineered for therapeutic and industrial applications. Here, we have reviewed comprehensively various synthetic biology techniques for engineering probiotic LAB strains, such as clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 mediated genome editing, homologous recombination, and recombineering. In addition, we also discussed heterologous protein expression systems used in engineering probiotic LAB. By combining computational biology with genetic engineering, there is a lot of potential to develop next-generation synthetic LAB with capabilities to address bottlenecks in industrial scale-up and complex biologics production. Recently, we started working on Lactochassis project where we aim to develop next generation synthetic LAB for biomedical application.

Novel Probiotic Lactic Acid Bacteria with In Vitro Bioremediation Potential of Toxic Lead and Cadmium.

In this study, newly isolated lactic acid bacteria (LAB) were screened for the potential bioremediation capacity against toxic lead (Pb, II) and cadmium (Cd, II) with their bioaccessibility and survivability. Five strains were selected from eighteen previously isolated probiotic LAB strains based on heavy metal-resistant potentiality through in vitro disc-diffusion assay. These five strains were evaluated in vitro to explore the Pb and Cd binding and removal efficiencies using Flame Atomic Absorption Spectrophotometry. At the same time, their bioaccessibility and survivability were assessed in a dynamic in vitro gastrointestinal digestion model. The results revealed that all the tested strains were shown to have a high magnitude of minimum inhibitory concentration values ranging from 500 to 2000mg/L with 5 to 25mm growth inhibition zones. The results also demonstrated a significant (P < 0.05) removal of Pb and Cd among five tested LAB strains. Lactobacillus delbrueckii subsp. bulgaricus LDMB02 showed the highest removal rates of Pb and Cd. It was also revealed that these strains significantly reduced Pb and Cd bioaccessibility from 42 to 50% and 40 to 58%, respectively. Moreover, these strains were shown to have significant survivability against Pb and Cd, ranging from 80.1 to 85.4% and 81.5 to 87.5%, respectively. This study recommends the immense potential exploit of LAB as a probiotic to protect human health from the adverse effects of Pb and Cd toxicity.

Novel Probiotic/Bacterial Cellulose Biocatalyst for the Development of Functional Dairy Beverage.

The development of innovative functional products with potential health benefits, under the concept of bio-economy, is flourishing. This study undertook an evaluation of non-dairy lactobacilli Lactiplantibacillus pentosus B329 and Lactiplantibacillus plantarum 820 as “ready to use” starter cultures. Lactic acid bacteria (LAB) cultures were evaluated for their fermentation efficiency, before and after freeze-drying, using cheese whey (CW) as a fermentation substrate and subsequent immobilization on bacteria cellulose (BC) to produce a novel biocatalyst. The biocatalyst was applied in functional sour milk production and compared with free cells via the assessment of physicochemical and microbiological properties and sensory evaluation. Evidently, LAB strains exhibited high fermentative activity before and after freeze-drying. Results of a 5-month storage stability test showed that viability was 19% enhanced by immobilization on BC, supporting the concept of “ready to use” cultures for the production of fermented beverages. Likewise, sour milk produced by the BC biocatalyst presented higher organoleptic scores, compared to the free cells case, whereas immobilization on BC enhanced probiotic viability during post-fermentation storage (4 °C, 28 days). The obtained high viability (>107 log cfu/g) demonstrated the efficacy of the proposed bioprocess for the production of functional/probiotic-rich beverages. Ultimately, this work presents a consolidated scheme that includes the advantages and the cooperative effect of probiotic LAB strains combined with a functional biopolymer (BC) towards the formulation of novel functional products that coincide with the pillars of food systems sustainability.

Potential for increasing folate contents of traditional African fermented sorghum gruel (Motoho) using presumptive probiotic lactic acid bacteria

Folate deficiency is commonly observed in most developing countries. Lactic acid bacteria (LAB) fermentation of cereal-based foods could be an alternative to improving folate intake. This study evaluated the possibility of improving the folate contents of traditional African fermented sorghum gruel (motoho) using indigenous potential probiotic LAB. A total of 220 LAB strains isolated from maize gruel were screened for extracellular and intracellular folate production. Strains were further examined for in vitro probiotic characteristics and antimicrobial activity. Sixteen (16) LAB strains exhibited high production of total folate which ranged between 44 and 180 µg/100 mL with the lowest and highest value in L. plantarum S8 and L. plantarum S49, respectively. Fermentation of sorghum gruel with folate producing probiotic LAB strains for in situ folate production in motoho was determined. The folate contents of sorghum motoho fermented with LAB ranged between 13 and 20 µg/100 mL while the control was below 2.5 µg/100 mL. Indigenous LAB strains from traditional African fermented gruel possess desirable in vitro probiotic properties and ability to produce folates, thus, could be used for natural in situ folate fortification. This study presents a strategy for future application of indigenous probiotic LAB cultures as natural means of fortifying cereal foods with folates.

Bioactivity of Biomass and Crude Exopolysaccharides Obtained by Controlled Submerged Cultivation of Medicinal Mushroom Trametes versicolor.

The aim of this study is to characterize the bioactivity of mycelial biomass and crude exopolysaccharides (EPS) produced by Trametes versicolor NBIMCC 8939 and to reveal its nutraceutical potential. The EPS (1.58 g/L) were isolated from a culture broth. The macrofungal biomass was rich in protein, insoluble dietary fibers and glucans. The amino acid composition of the biomass was analyzed and 18 amino acids were detected. Three mycelial biomass extracts were prepared and the highest total polyphenol content (16.11 ± 0.14 mg GAE/g DW) and the total flavonoid content (5.15 ± 0.03 mg QE/g DW) were found in the water extract. The results indicated that the obtained EPS were heteropolysaccharides with glucose as the main building monosaccharide and minor amounts of mannose, xylose, galactose, fucose and glucuronic acid. Fourier Transform Infrared Spectroscopy (FTIR) confirmed the complex structure of the crude EPS. Five probiotic lactic acid bacteria strains were used for the determination of the prebiotic effect of the crude EPS. The anti-inflammatory potential was tested in vitro using cell line HT-29. The significant decrease of IL-1 and IL-8 and increase of TGF-beta expression revealed anti-inflammatory potential of the crude exopolysaccharides from T. versicolor.

Peptidomic Characteristic of Peptides Generated in Dry-Cured Loins with Probiotic Strains of LAB during 360-Days Aging

Peptidomics refers to the comprehensive profiling of endogenous peptides obtained from biological sources. The formation of endogenous peptides is dependent on not only endogenous factors but also exogenous factors such as microbial proteases or process conditions, including fermentation. This study analyzed the probiotic strains of Lactobacillus rhamnosus LOCK900 (LOCK), Bifidobacterium animalis ssp. lactis BB-12 (BB12), and potential probiotic Lactobacillus acidophilus Bauer Ł0938 (BAUER) to assess their ability of fermentation and peptide production in dry-cured pork loin. The peptides obtained after in vitro digestion were characterized by liquid chromatography–tandem mass spectrometry. Based on the sequences identified, the degree of similarity or differences between the peptides was determined and presented graphically on the factor plane. The charts showed that the meat products aged for 180 and 270 days were the most diverse when BB12 or BAUER were used as starter cultures. Myosin and keratin were identified as the most likely precursors of bioactive peptides in products obtained using this strain of lactic acid bacteria (LAB). The knowledge acquired from this study may contribute to the design of functional meat products as the results revealed not only the peptidogenic potential of the LAB strains indicated on their beneficial effect on the bioactivity of peptides.

Application of Potentially Probiotic Fruit-Derived Lactic Acid Bacteria Loaded into Sodium Alginate Coatings to Control Anthracnose Development in Guava and Mango During Storage.

This study evaluated the efficacy of potentially probiotic fruit-derived lactic acid bacteria (LAB) strains loaded into sodium alginate (SA) coatings to control the anthracnose development in guava cv. Paluma and mango cv. Palmer caused by distinct pathogenic Colletotrichum species (C. asianum, C. fructicola, C. tropicale, C. siamense, C. karstii, and C. gloeosporioides) during 15days of room temperature storage (25 ± 0.5°C). The effects of the formulated coatings on physicochemical parameters indicative of overall postharvest quality of guava and mango were evaluated. The eight examined LAB strains caused strong inhibition on the mycelial growth of all target Colletotrichum species in vitro. LAB strains with the highest inhibitory effects (Levilactobacillus brevis 59, Lactiplantibacillus pentosus 129, and Limosilactobacillus fermentum 263) on the target Colletotrichum species were incorporated into SA coatings. These strains had viable counts of > 6 log CFU/mL in SA coatings during 15days of room temperature storage. Application of coatings with SA + L. brevis 59, SA + L. pentosus 129, and SA + L. fermentum 263 delayed the development and decreased the severity of anthracnose lesions in guava and mango artificially contaminated with either of the tested Colletotrichum species. These coatings impacted positively on some physicochemical parameters indicative of postharvest quality and more prolonged storability of guava and mango. The formulated SA coatings loaded with tested fruit-derived potentially probiotic LAB strains could be innovative and effective strategies to control postharvest anthracnose and extend the storability of guava and mango.

In-Vitro Characterization of Growth Inhibition against the Gut Pathogen of Potentially Probiotic Lactic Acid Bacteria Strains Isolated from Fermented Products.

Lactic acid bacteria (LAB) are probiotic candidates that may restore the balance of microbiota populations in intestinal microbial ecosystems by controlling pathogens and thereby promoting host health. The goal of this study was to isolate potential probiotic LAB strains and characterize their antimicrobial abilities against pathogens in intestinal microbiota. Among 54 LAB strains isolated from fermented products, five LAB strains (NSMJ15, NSMJ16, NSMJ23, NSMJ42, and NFFJ04) were selected as potential probiotic candidates based on in vitro assays of acid and bile salt tolerance, cell surface hydrophobicity, adhesion to the intestinal epithelium, and antagonistic activity. Phylogenetic analysis based on 16S rRNA genes showed that they have high similarities of 99.58–100% to Lacticaseibacillus paracasei strains NSMJ15 and NFFJ04, Lentilactobacillus parabuchneri NSMJ16, Levilactobacillus brevis NSMJ23, and Schleiferilactobacillus harbinensis NSMJ42. To characterize their antimicrobial abilities against pathogens in intestinal microbiota, the impact of cell-free supernatant (CFS) treatment in 10% (v/v) fecal suspensions prepared using pooled cattle feces was investigated using in vitro batch cultures. Bacterial community analysis using rRNA amplicon sequencing for control and CFS-treated fecal samples at 8 and 16 h incubation showed the compositional change after CFS treatment for all five LAB strains. The changed compositions were similar among them, but there were few variable increases or decreases in some bacterial groups. Interestingly, as major genera that could exhibit pathogenicity and antibiotic resistance, the members of Bacillus, Escherichia, Leclercia, Morganella, and Vagococcus were decreased at 16 h in all CFS-treated samples. Species-level classification suggested that the five LAB strains are antagonistic to gut pathogens. This study showed the probiotic potential of the five selected LAB strains; in particular, their antimicrobial properties against pathogens present in the intestinal microbiota. These strains would therefore seem to play an important role in modulating the intestinal microbiome of the host.

Novel fructooligosaccharides of Dioscorea alata L. tuber have prebiotic potentialities

Dioscorea alata L. (Dioscoreaceae), commonly known as water yam, is a popular tuber crop vegetable used across the globe. It is mainly composed of starch (75–84% of the dry weight) with a small amount of proteins, lipids, and vitamins and is very rich in minerals. It has nutritional superiority compared with other tropical root crops. The main aim of the study was to extract and characterize the fructooligosaccharides (FOSs) fractions from the tuber of this plant and evaluate their prebiotic properties. The soluble FOSs were fractionated by the conventional solid-phase extraction method and were characterized by FT-IR, HPTLC, and ESI–MS analyses. In addition, the beneficial health attributes of these fractions were checked for in vitro and in vivo prebiotic properties in Swiss albino mice. The FT-IR, HPTLC, and ESI–MS analyses of the FOSs from the hot water and 80% ethanol fractions showed that both are rich in inulin-like prebiotics. Furthermore, these FOSs showed an excellent prebiotic activity score and had enhanced in vitro cholesterol-binding activities in the presence of probiotic lactic acid bacteria (LAB) strains. Moreover, these had good antioxidant efficacies with IC50 values of 36.51 µg/ml and 33.12 µg/ml. In vivo treatment in Swiss albino mice, these FOSs had significantly lowered serum cholesterol (df = 6, p value < 2.2e−16), serum HDL (df = 6, p value = 3.68e−11), serum LDL (df = 6, p value < 2e−16), serum VLDL (df = 6, p value = 2.2e−13), serum triglycerides (df = 6, p value < 2e−16), blood sugar (df = 6, p value = 0.01857), serum glutamic pyruvic transaminase (S.G.P.T., df = 6, p value = 0.01189), serum glutamic oxaloacetic transaminase (S.G.O.T., df = 6, p value = 0.01239) but no significant difference found in weight gain (df = 6, p value = 0.166) and serum creatinine (df = 6, p value = 0.987). Furthermore, they enhanced IgA-mediated immunomodulation and influenced the gut colonization of probiotic LAB strains than the standard prebiotics, inulin. Thus, the study revealed that the FOSs from the storage tuber of D. alata possess some novel prebiotic that could contribute to human nutrition. This is the first report of the prebiotic FOS characterization from this plant.

Screening beneficial bacteriostatic lactic acid bacteria in the intestine and studies of bacteriostatic substances.

Lactic acid bacteria (LAB) are a representative probiotic. As the dominant flora in the human intestinal tract, LAB can regulate the balance of human intestinal flora and improve host health. The purpose of this study was to isolate and screen LAB that are well suited to the intestinal characteristics of the Chinese population, with excellent probiotics and high antibacterial activity. After 16S ribosomal RNA (rRNA) homology and phylogenetic tree analysis, potential probiotics were tested for their antibacterial activity, resistance to artificial gastrointestinal fluid and drugs, surface hydrophobicity, and safety. Three strains of LAB with acid resistance, bile salt resistance, epithelial cell adhesion, and no multidrug resistance were selected: Lactobacillus salivarius, Leuconostoc lactis, and Lactobacillus paracasei. Analysis of the antibacterial active substances in the three strains and their fermentation broths revealed that the main antibacterial substances of L. lactis were organic acids, whereas those of L. salivarius and L. paracasei were organic acids and bacteriocins with broad-spectrum antibacterial activity. These three strains of probiotic LAB with high antibacterial activity were identified as bacterial resources that could potentially be used to develop probiotic preparations for the prevention and treatment of intestinal diseases caused by intestinal pathogens.

Fermentation Enhanced Biotransformation of Compounds in the Kernel of Chrysophyllum albidum.

Chrysophyllum albidum Linn (African star apple) is a fruit with extensive nutritional and medicinal benefits. The fruit and kernel in the seed are both edible. Strains of lactic acid bacteria (LAB) were isolated from fermented seeds and assessed for probiotic characteristics. The extracts in both the unfermented and the fermented aqueous extracts from the kernels obtained from the seeds of C. albidum were subjected to analysis using the gas chromatography/mass spectrometry (GC-MS) method. This analysis identified the bioactive compounds present as possible substrate(s) for the associated organisms inducing the fermentation and the resultant biotransformed products formed. Three potential probiotic LAB strains identified as Lactococcus raffinolactis (ProbtA1), Lactococcus lactis (ProbtA2a), and Pediococcus pentosaceus (ProbtA2b) were isolated from the fermented C. albidum seeds. All strains were non hemolytic, which indicated their safety, Probt (A1, A2a, and A2b) grew in an acidic environment (pH 3.5) during the 48-h incubation time, and all three strains grew in 1% bile, and exhibited good hydrophobicity and auto-aggregation properties. Mucin binding proteins was not detected in any strain, and bile salt hydrolase was detected in all the strains. l-lactic acid (28.57%), norharman (5.07%), formyl 7E-hexadecenoate (1.73%), and indole (1.51%) were the four major constituents of the fermented kernel of the C. albidum, while 2,5-dimethylpyrazine (C1, 1.27%), 3,5-dihydroxy-6-methyl-2,3-dihydropyran-4-one (C2, 2.90%), indole (C3, 1.31%), norharman (C4, 3.01%), and methyl petroselinate (C5, 4.33%) were the five major constituents of the unfermented kernels. The isolated LAB are safe for consumption. The fermenting process metabolized C1, C2, and C5, which are possible starter cultures for the growth of probiotics. Fermentation is an essential tool for bioengineering molecules in foods into safe and health beneficial products.

Intramammary inoculation with lactic acid bacteria at dry-off triggers an immunomodulatory response in dairy cows.

The use of antibiotics to prevent bovine mastitis is responsible for the emergence and selection of resistant strains. Lactic acid bacteria (LAB) could be introduced into animal feed as an alternative prevention method that would bypass the risk of resistance development. In previous research, we demonstrated that two probiotic LAB strains isolated from bovine milk were capable of stimulating the production of antibodies and the host's immune cellular response in the udder. The present study aimed to elucidate whether the antibodies of animals inoculated with these strains were able to increase phagocytosis by neutrophils and inhibit the growth of different mastitis-causing pathogens. Moreover, the effect of LAB on the expression of pro-inflammatory cytokines was assessed. Ten animals were inoculated intramammarily with 106 cells of the two strains at dry-off. The blood serum was tested for its ability to opsonize bovine mastitis pathogens, the in vitro bactericidal activity of bovine blood and milk against these pathogens was determined, and cytokine mRNA expression was quantified in milk somatic cells. The inoculated animals did not show abnormal signs of sensitivity to the LAB. Their blood serum significantly enhanced the phagocytosis of Staphylococcus spp. and the LAB. Escherichia coli and Streptococcus uberis were inhibited by the milk serum but not the blood serum, whereas Staphylococcus aureus and Staphylococcus haemolyticus were inhibited by both. In regard to cytokine expression, interleukin (IL)-1β increased markedly for up to 4 h post-inoculation, and an increase in IL-8 was observed 4, 12 and 24 h after inoculation. Tumour necrosis factor-α mRNA increased 1 and 2 h after inoculation and a significant difference was registered at 6 h for interferon-γ. This rapid immunomodulatory response shows that inoculating animals with LAB at dry-off, when they are especially susceptible, could be a useful strategy for the prevention of bovine mastitis.

Isolation, characterization and selection of potential probiotic lactic acid bacteria from feces of wild boar, native pig and commercial pig

In livestock production, lactic acid bacteria (LAB) play an important role in improving performance, mitigating diseases due to their ability to maintain the balance of gastrointestinal microbes and resistance to pathogenic bacteria. As resident adapted microorganisms, LAB should be isolated and characterized from the homologous host to improve their efficiency as additives in pig feeding. In present study, cultivable LAB numbers from commercial pig (CP), native pig (NP) and wild boar (WB) samples ranged from 4.64 to 7.39 (log10 CFU/g), and the LAB counts in commercial pig and native pig feces were much higher than wild boar. Ninety-six LAB strains were detected in commercial pig, native pig and wild boar feces, being represented by thirteen different species of Enterococcus, Lactobacillus, Pediococcus and Weissella genera. Tests in vitro were conducted including survival in simulated gastrointestinal tract conditions, antimicrobial activity, organic acid production, hydrophobicity, auto-aggregation, Caco-2 surface adhesion, haemolytic activity and antibiotic resistance. According to the results of acid and bile salt resistant and bacteriostatic test, twelve LAB strains were selected as potential probiotics and their cell surface properties were further evaluated. Based on multivariate principal component analysis, nine probiotic LAB strains (Lactobacillus plantarum NCUA001005 and NCUA001014, Pediococcus pentosaceus NCUA006009, Lactobacillus reuteri NCUA064001 and NCUA064006, Lactobacillus johnsonii NCUA063001, NCUA063006 and NCUA063008, Lactobacillus casei NCUA011001) showed better characteristics and might be selected to explore their potential use as probiotics. These strains were good candidates for further investigation in vivo studies to assess their potential health benefits as feed supplements.

Sustainable microbial cell nanofactory for zinc oxide nanoparticles production by zinc-tolerant probiotic Lactobacillus plantarum strain TA4

BackgroundThe use of microorganisms in the biosynthesis of zinc oxide nanoparticles (ZnO NPs) has recently emerged as an alternative to chemical and physical methods due to its low-cost and eco-friendly method. Several lactic acid bacteria (LAB) have developed mechanisms in tolerating Zn2+ through prevention against their toxicity and the production of ZnO NPs. The LAB’s main resistance mechanism to Zn2+ is highly depended on the microorganisms’ ability to interact with Zn2+ either through biosorption or bioaccumulation processes. Besides the inadequate studies conducted on biosynthesis with the use of zinc-tolerant probiotics, the understanding regarding the mechanism involved in this process is not clear. Therefore, this study determines the features of probiotic LAB strain TA4 related to its resistance to Zn2+. It also attempts to illustrate its potential in creating a sustainable microbial cell nanofactory of ZnO NPs.ResultsA zinc-tolerant probiotic strain TA4, which was isolated from local fermented food, was selected based on the principal component analysis (PCA) with the highest score of probiotic attributes. Based on the 16S rRNA gene analysis, this strain was identified as Lactobacillus plantarum strain TA4, indicating its high resistance to Zn2+ at a maximum tolerable concentration (MTC) value of 500 mM and its capability of producing ZnO NPs. The UV–visible spectroscopy analysis proved the formations of ZnO NPs through the notable absorption peak at 380 nm. It was also found from the dynamic light scattering (DLS) analysis that the Z-average particle size amounted to 124.2 nm with monodisperse ZnO NPs. Studies on scanning electron microscope (SEM), energy-dispersive X-ray (EDX) spectroscopy, and Fourier-transform infrared spectroscopy (FT-IR) revealed that the main mechanisms in ZnO NPs biosynthesis were facilitated by the Zn2+ biosorption ability through the functional groups present on the cell surface of strain TA4.ConclusionsThe strong ability of zinc-tolerant probiotic of L. plantarum strain TA4 to tolerate high Zn2+ concentration and to produce ZnO NPs highlights the unique properties of these bacteria as a natural microbial cell nanofactory for a more sustainable and eco-friendly practice of ZnO NPs biosynthesis.

Suppressive effects of Streptococcus thermophilus KLDS 3.1003 on some foodborne pathogens revealed through in vitro, in vivo and genomic insights.

Foodborne diseases (FBDs) remain a persistent global challenge and recent research efforts suggest that lactic acid bacteria (LAB) strains can contribute towards their prevention and treatment. This study investigates the genetic properties of Streptococcus thermophilus KLDS 3.1003 as a potential probiotic and health-promoting LAB strain as well as its in vitro and in vivo activities against two foodborne pathogens. In vitro, its antimicrobial activities and tolerance levels in simulated bile salts and acids were determined. The cytotoxic effects of the LAB strain in RAW264.7 cells were also evaluated. For in vivo evaluation, 24 BALB/c mice were orally administered control and trial diets for 14 days. Genomic analyses of this strain's bacteriocin configuration, stress response system and multidrug resistance genes were annotated to validate in vitro and in vivo results. In vitro antimicrobial results show that the cells and CFS of S. thermophilus KLDS 3.1003 could inhibit both pathogens with the former being more effective (P < 0.05). In addition, its cell-free supernatant (CFS) could inhibit the growth of both pathogens, with catalase treatment having the highest effect against it. More so, after 3 h of incubation, survivability levels of S. thermophilus KLDS 3.1003 were significantly high (P < 0.05). LPS-induced RAW264.7 cell activities were also significantly reduced by 108-109 CFU mL-1 of S. thermophilus KLDS. In vivo, significant weight losses were inhibited in the TSTEC group compared to the TSTSA group (P < 0.05). Moreover, pathogen-disrupted blood biochemical parameters like HDL, LDL, TP, TG, AST, ALT and some minerals were restored in the respective prevention groups (TSTEC and TSTSA). Genomic analyses showed that S. thermophilus KLDS 3.1003 has bacteriocin-coding peptides, which accounts for its antimicrobial abilities in vitro and in vivo. S. thermophilus KLDS 3.1003 is also endowed with intact genes for acid tolerance, salt-resistance, cold and heat shock responses and antioxidant activities, which are required to promote activities against the selected foodborne pathogens. This study showed that S. thermophilus KLDS 3.1003 has the genomic capacity to inhibit foodborne pathogens' growth in vitro and in vivo, thus qualifying it as a potential probiotic, antimicrobial and bio-therapeutic candidate.

α-Glucosidase and ACE dual inhibitory protein hydrolysates and peptide fractions of sprouted quinoa yoghurt beverages inoculated with Lactobacillus casei

Dietary protein peptides from quinoa yoghurt beverage (QYB) fermented with probiotic lactic acid bacteria strains play a protective role against diabetes and hypertension. In this study, the α-glucosidase and ACE inhibitory activities of germination-based protein hydrolysates of QYB were investigated. All protein hydrolysates exhibited a dose and strain-dependent inhibition on the enzymes. The inhibition of α-glucosidase was the highest in QLCSY13 (IC50 = 8.86 mg/mL), while ACE inhibition was the highest in QLCZ (IC50 = 0.03 mg/mL). Overall, QLCSY13 had the highest inhibitory activities, which was ascribed to its relatively higher amino acid contents and hydrophobicity. In addition, the ACE and α-glucosidase inhibitory activities of peptide fractions identified by RP-HPLC were 127 ± 4.29 mg/mL and 10.39 ± 4.73 mg/mL respectively. Among the potent inhibitory peptide sequences identified, both LAHMIVAGA and VAHPVF significantly had α-glucosidase and ACE inhibitory activities. Consequently, dietary protein peptides present in QYB had anti-hypertensive and anti-diabetic potentials.