Antagonistic properties of Lactiplantibacillus plantarum MYSVB1 against Alternaria alternata: a putative probiotic strain isolated from the banyan tree fruit.

Alternaria alternata, a notorious phytopathogenic fungus, has been documented to infect several plant species, leading to the loss of agricultural commodities and resulting in significant economic losses. Lactic acid bacteria (LAB) hold immense promise as biocontrol candidates. However, the potential of LABs derived from fruits remains largely unexplored. In this study, several LABs were isolated from tropical fruit and assessed for their probiotic and antifungal properties. A total of fifty-five LABs were successfully isolated from seven distinct fruits. Among these, seven isolates showed inhibition to growth of A. alternata. Two strains, isolated from fruits: Ficus benghalensis, and Tinospora cordifolia exhibited promising antifungal properties against A. alternata. Molecular identification confirmed their identities as Lactiplantibacillus plantarum MYSVB1 and MYSVA7, respectively. Both strains showed adaptability to a wide temperature range (10-45°C), and salt concentrations (up to 7%), with optimal growth around 37 °C and high survival rates under simulated gastrointestinal conditions. Among these two strains, Lpb. plantarum MYSVB1 demonstrated significant inhibition (p < 0.01) of the growth of A. alternata. The inhibitory effects of cell-free supernatant (CFS) were strong, with 5% crude CFS sufficient to reduce fungal growth by >70% and complete inhibition by 10% CFS. Moreover, the CFS was inhibitory for both mycelial growth and conidial germination. CFS retained its activity even after long cold storage. The chromatographic analysis identified organic acids in CFS, with succinic acid as the predominant constituent, with lactic acid, and malic acid in descending order. LAB strains isolated from tropical fruits showed promising probiotic and antifungal properties, making them potential candidates for various applications in food and agriculture.

Antifungal activity of probiotic strain Lactiplantibacillus plantarum MYSN7 against Trichophyton tonsurans.

The primary objective of this study was to assess the probiotic attributes and antifungal activity of lactic acid bacteria (LAB) against the fungus, Trichophyton tonsurans. Among the 20 isolates screened for their antifungal attributes, isolate MYSN7 showed strong antifungal activity and was selected for further analysis. The isolate MYSN7 exhibited potential probiotic characteristics, having 75 and 70% survival percentages in pH3 and pH2, respectively, 68.73% tolerance to bile, a moderate cell surface hydrophobicity of 48.87%, and an auto-aggregation percentage of 80.62%. The cell-free supernatant (CFS) of MYSN7 also showed effective antibacterial activity against common pathogens. Furthermore, the isolate MYSN7 was identified as Lactiplantibacillus plantarum by 16S rRNA sequencing. Both L. plantarum MYSN7 and its CFS exhibited significant anti-Trichophyton activity in which the biomass of the fungal pathogen was negligible after 14 days of incubation with the active cells of probiotic culture (106 CFU/ml) and at 6% concentration of the CFS. In addition, the CFS inhibited the germination of conidia even after 72 h of incubation. The minimum inhibitory concentration of the lyophilized crude extract of the CFS was observed to be 8 mg/ml. Preliminary characterization of the CFS showed that the active component would be organic acids in nature responsible for antifungal activity. Organic acid profiling of the CFS using LC-MS revealed that it was a mixture of 11 different acids, and among these, succinic acid (9,793.60 µg/ml) and lactic acid (2,077.86 µg/ml) were predominant. Additionally, a scanning electron microscopic study revealed that CFS disrupted fungal hyphal structure significantly, which showed scanty branching and bulged terminus. The study indicates the potential of L. plantarum MYSN7 and its CFS to control the growth of T. tonsurans. Furthermore, in vivo studies need to be conducted to explore its possible applications on skin infections.

Exploring biotechnological and functional characteristics of probiotic yeasts: A review.

Probiotics are vital and beneficial organisms which offers the health benefits to the host organisms. The fungal probiotic field is one of the developing fields nowadays. Yeast has an enormous and diverse group of microorganisms that is attracting and expanding the attention from researchers and industries. Saccharomyces boulardii, the only patented strain belonging to yeast genera for the human use, has been broadly evaluated for its probiotic effect. Yeasts belonging to the genera Debaryomyces, Pichia, Yarrowia, Meyerozyma, Kluyveromyces etc.., have attained more interest because of their beneficial and probable probiotic features. These yeast probiotics produce VOCs (Volatile organic compounds), mycocins and antimicrobials which shows the antagonistic effect against pathogenic fungi and bacteria. Additionally, those yeasts have been recorded as good plant growth promoting microorganisms. Yeast has an important role in environmental applications such as bioremediation and removal of metals like chromium, mercury, lead etc., from waste water. Probiotic yeasts with their promising antimicrobial, antioxidant, anticancer properties, cholesterol assimilation and immunomodulatory effects can also be utilized as biotherapeutics. In this review article we have made an attempt to address important yeast probiotic attributes.

Probiotic properties of Lactobacillus casei - MYSRD 108 and Lactobacillus plantarum-MYSRD 71 with potential antimicrobial activity against Salmonella paratyphi.

A total of 130 isolates were screened, twelve isolates were characterized for probiotic attributes and two isolates with best probiotic features were evaluated in the study. Isolates MYSRD108 and MYSRD71 survived gastric conditions and were susceptible to tested antibiotics. Isolates showed more vital cell surface traits such as autoaggregation of 89.2 and 88.5% and cell surface hydrophobicity of 61 and 64%. PCR amplification followed by 16sRNA sequencing results confirmed that the isolates as Lactobacillus casei (MYSRD 108) and Lactobacillus plantarum (MYSRD 71). During this study, the Cells and their Cell Free Supernatant (CFS) were examined for antimicrobial activity. Both the isolates inhibited different bacterial pathogens in which the growth of S. paratyphi was significantly reduced. Further, their CFS also showed inhibitory effects against S. paratyphi with agar well diffusion and Minimum Inhibitory Concentration using Broth micro dilution method. The antimicrobial compounds in the CFS was characterized to different constraints such as pH neutralization, heat treatment, Hydrogen peroxide test and storage stability at -20> °C and represented that the antagonistic acitivity against Salmonella is due to the presence of organic acids in the supernatants that lowered the pH. These strains were further examined for the inhibition of S. paratyphi biofilm. The results indicated that CFS reduced S. paratyphi biofilm by more than 75% and the number of Salmonella biofilm was effectively reduced using 15% concentration of CFS. These strains may be used to produce antimicrobial compounds which can be a substitute for chemical preservatives in food industry.

Probiotic and Antifungal Attributes of Levilactobacillus brevis MYSN105, Isolated From an Indian Traditional Fermented Food Pozha.

The use of probiotics and antifungal capabilities of the lactic acid bacteria (LAB) isolated from different niches is a strategy to prepare functional cultures and biopreservatives for food/feed industries. In the present study, LAB strains isolated from an Indian traditional fermented food, Pozha, were evaluated for their probiotic properties and biocontrol potential. A total of 20 LAB isolates were selected from Pozha samples collected aseptically and screened for their antagonistic activity against Fusarium verticillioides. Among the bioactive isolates, Lacticaseibacillus brevis MYSN105 showed the highest antifungal activity in vitro, causing some morphological alterations such as damaged mycelia and deformed conidia. Cell-free supernatant (CFS) from L. brevis MYSN105 at 16% concentration effectively reduced the mycelial biomass to 0.369 g compared to 1.938 g in control. Likewise, the conidial germination was inhibited to 20.12%, and the seed treatment using CFS induced a reduction of spore count to 4.1 × 106 spores/ml compared to 1.1 × 109 spores/ml for untreated seeds. The internal transcribed spacer (ITS) copy number of F. verticillioides decreased to 5.73 × 107 and 9.026 × 107 by L. brevis MYSN105 and CFS treatment, respectively, compared to 8.94 × 1010 in control. The L. brevis MYSN105 showed high tolerance to in vitro gastrointestinal conditions and exhibited high adhesive abilities to intestinal epithelial cell lines. The comparative genome analysis demonstrated specific secondary metabolite region coding for bacteriocin and T3PKS (type III polyketide synthase) possibly related to survival and antimicrobial activity in the gut environment. Our results suggest that L. brevis MYSN105 has promising probiotic features and could be potentially used for developing biological control formulations to minimize F. verticillioides contamination and improve food safety measures.

Probiotic Properties of Lactic Acid Bacteria Isolated From Neera: A Naturally Fermenting Coconut Palm Nectar.

Probiotic bacteria were isolated from different traditional fermented foods as there are several such foods that are not well explored for their probiotic activities. Hence, the present study was conducted to find the potential of lactic acid bacteria (LAB) as probiotics that were isolated from the sap extract of the coconut palm inflorescence - Neera, which is a naturally fermented drink consumed in various regions of India. A total of 75 isolates were selected from the Neera samples collected aseptically in the early morning (before sunrise). These isolates were initially screened for cultural, microscopic, and biochemical characteristics. The initial screening yielded 40 Gram-positive, catalase-negative isolates that were further subjected to acid - bile tolerance with resistance to phenol. Among 40 isolates, 16 survived screening using analysis of cell surface hydrophobicity, auto aggregation with adhesion to epithelial cells, and gastric-pancreatic digestion for gastrointestinal colonization. The isolates were also assessed for antimicrobial, antibiotic sensitivity, and anti-oxidative potential. The safety of these isolates was evaluated by their hemolytic and deoxyribonuclease (DNase) activities. Based on these results, seven isolates with the best probiotic attributes were selected and presented in this study. These LAB isolates, with 51.91-70.34% survival at low pH, proved their resistance to gastric conditions. The cell surface hydrophobicity of 50.32-77.8% and auto aggregation of 51.02-78.95% represented the adhesion properties of these isolates. All the seven isolates exhibited good antibacterial and antifungal activity, showing hydroxyl-scavenging activity of 32.86-77.87%. The results proved that LAB isolated from Neera exhibited promising probiotic properties and seem favorable for use in functional fermented foods as preservatives.

Lactobacillus plantarum MYS6 Ameliorates Fumonisin B1-Induced Hepatorenal Damage in Broilers.

Fumonisin B1 (FB1), a mycotoxin produced by Fusarium species is a predominant Group 2B carcinogen occurring in maize and maize-based poultry feeds. It is shown to be nephrotoxic, hepatotoxic, neurotoxic, and immunosuppressing in animals. In this study, we report the ameliorating effects of a probiotic strain, Lactobacillus plantarum MYS6 on FB1-induced toxicity and oxidative damage in broilers. A 6-week dietary experiment consisting of 48 broilers was performed in six treatment groups. Probiotic treatment (109 cells/mL) involved pre-colonization of broilers with L. plantarum MYS6 while co-administration treatment involved supplementation of probiotic and FB1-contaminated diet (200 mg/Kg feed) simultaneously. At the end of the treatment period, growth performance, hematology, serum biochemistry, and markers of oxidative stress in serum and tissue homogenates were evaluated in all the broilers. The histopathological changes in hepatic and renal tissues were further studied. The results demonstrated that administration of L. plantarum MYS6 efficiently improved the feed intake, body weight and feed conversion ratio in broilers. It mitigated the altered levels of hematological indices such as complete blood count, hemoglobin, and hematocrit. Serum parameters such as serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, creatinine, cholesterol, triglycerides, and albumin were significantly restored after administering the probiotic in FB1-intoxicated broilers. Additionally, L. plantarum MYS6 alleviated the levels of oxidative stress markers in serum and tissue homogenate of liver. The histopathological data of liver and kidney further substantiated the overall protection offered by L. plantarum MYS6 against FB1-induced cellular toxicity and organ damage in broilers. Our results indicated that co-administration of probiotic along with the toxin had better effect in detoxification compared to its pre-colonization in broilers. Collectively, our study signifies the protective role of L. plantarum MYS6 in ameliorating the FB1-induced toxicity in the vital organs and subsequent oxidative stress in broilers. The probiotic L. plantarum MYS6 can further be formulated into a functional feed owing to its anti-fumonisin attributes and role in mitigating FB1-induced hepatorenal damage.