Mayakrishnan Vijayakumar

In-vitro assessment of the probiotic potential of Lactobacillus plantarum KCC-24 isolated from Italian rye-grass (Lolium multiflorum) forage

The aim of the present study was to determine the probiotic potential of the lactic acid bacteria Lactobacillus plantarum KCC-24 (L. plantarum KCC-24), that was isolated and characterized from Italian ryegrass (Lolium multiflorum) forage. The following experiments were performed to assess the probiotic characteristics such as antifungal activity, antibiotic susceptibility, resistance to low pH, stimulated gastric juice and bile salts, proteolytic activity, auto-aggregation, cell surface hydrophobicity, and in vitro antioxidant property. The isolated L. plantarum KCC-24 exhibited significant antifungal activity against the various fungal strains of Aspergillus fumigatus (73.43%), Penicillium chrysogenum (59.04%), Penicillium roqueforti (56.67%), Botrytis elliptica (40.23%), Fusarium oxysporum (52.47%) and it was susceptible to numerous antibiotics, survived in low pH, was resistant to stimulated gastric juices and bile salts (0.3% w/v). Moreover, L. plantarum KCC-24 exhibited good proteolytic activity. In addition L. plantarum KCC-24 showed potent antioxidant and hydrogen peroxide resistant property. In conclusion, the isolated L. plantarum KCC-24 exhibited several characteristics to prove its excellent as a potential probiotic candidate for developing quality food for ruminant animals and human.

Assessment of probiotic, antifungal and cholesterol lowering properties of Pediococcus pentosaceus KCC-23 isolated from Italian ryegrass

BACKGROUND Lactic acid bacteria (LAB) are important for the processing of various food products. Although genetically modified organisms have contributed to improvements in various food products, there are some limitations. Thus, the discovery of wild strains from natural sources must be considered as the most suitable approach for identifying new LAB. Therefore, we planned to isolate and characterise the LAB from Italian ryegrass forage and evaluate their biological potential. RESULTS A total of 28 strains were isolated and screened for their anti-fungal and probiotic properties. A single strain was selected due to its antifungal and probiotic efficiency. The strain was identified as Pediococcus pentosaceus KCC-23. The strain KCC-23 showed effective inhibition against Aspergillus fumigatus, Pencillium chrysogenum, Pencillium roqueforti, Botrytis elliptica and Fusarium oxysporum. Further, it survived low pH, and the presence of bile salts and gastric juice. It exhibited significant aggregation and hydrophobicity properties. The KCC-23 effectively assimilated cholesterol and had the ability to utilise pre-biotics such as raffinose and inulin. Finally, KCC-23 exhibited significant free radical scavenging activity. CONCLUSION P. pentosaceus KCC-23 showed effective anti-fungal, probiotic and anti-oxidant properties and would be a promising isolate for exploitation in the formulation of food for ruminants and humans.

Probiotic Potential of Lactobacillus Strains with Antifungal Activity Isolated from Animal Manure

The aim of the study was to isolate and characterize the lactic acid bacteria (LAB) from animal manure. Among the thirty LAB strains, four strains, namely, KCC-25, KCC-26, KCC-27, and KCC-28, showed good cell growth and antifungal activity and were selected for further characterization. Biochemical and physiology properties of strains confirmed that the strains are related to the Lactobacillus sp.; further, the 16S rRNA sequencing confirmed 99.99% sequence similarity towards Lactobacillus plantarum. The strains exhibited susceptibility against commonly used antibiotics with negative hemolytic property. Strains KCC-25, KCC-26, KCC-27, and KCC-28 showed strong antifungal activity against Aspergillus fumigatus, Penicillium chrysogenum, Penicillium roqueforti, Botrytis elliptica, and Fusarium oxysporum, respectively. Fermentation studies noted that the strains were able to produce significant amount of lactic, acetic, and succinic acids. Further, the production of extracellular proteolytic and glycolytic enzymes, survival under low pH, bile salts, and gastric juice together with positive bile salt hydrolase (Bsh) activity, cholesterol lowering, cell surface hydrophobicity, and aggregation properties were the strains advantages. Thus, KCC-25, KCC-26, KCC-27, and KCC-28 could have the survival ability in the harsh condition of the digestive system in the gastrointestinal tract. In conclusion, novel L. plantarum KCC-25, KCC-26, KCC-27, and KCC-28 could be considered as potential antimicrobial probiotic strains.

Lactobacillus plantarum Improves the Nutritional Quality of Italian Ryegrass with Alfalfa Mediated Silage

ABSTRACT The present study was planned to analyze the nutritional quality, microbial counts and fermentative acids in Italian ryegrass (IRG) 80% and alfalfa 20% (IRG-HV) mediated silage inoculated with lactic acid bacteria (LAB) as a probiotic strain for 3 months. Crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF), total digestible nutrient (TDN) and In- vitro dry matter digestibility (IVDMD), lactic acid bacteria (LAB), yeast and fungi counts and fermentation metabolites such as lactic acid, acetic acid and butyric acids were analyzed. The result shows that the nutritional quality and metabolite profiles of silage were significantly improved with LAB. For microbial counts, LAB showed dominant followed by yeast as compared with control silage. The pH of the silage also reduced significantly when silage inoculated with LAB. The result confirmed that silage preparation using different crops with L. plantarum inoculation is most beneficial for the farmers.(

Trigonelline protects the cardiocyte from hydrogen peroxide induced apoptosis in H9c2 cells

OBJECTIVE To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline (TG) for reducing the H2O2 induced toxicity in H9c2 cells. METHODS Cytotoxicity and antioxidant activity of TG was assessed by EZ-CYTOX kit. RNA extraction and cDNA synthesized according to the kit manufacture protocol. Apoptosis was measured by the Flowcytometry, general PCR and qPCR. RESULTS It was found that the TG significantly rescued the morphology of the H9c2 cells. Treatment of cells with TG attenuated H2O2 induced cell deaths and improved the antioxidant activity. In addition, TG regulated the apoptotic gene caspase-3, caspase-9 and anti-apoptotic gene Bcl-2, Bcl-XL during H2O2 induced oxidative stress in H9c2 cells. These results were comparable with quercetin treatment. For evident, flow cytometer results also confirmed the TG significantly reduced the H2O2 induced necrosis and apoptosis in H9c2 cells. However, further increment of TG concentration against H2O2 could induce the necrosis and apoptosis along with H2O2. CONCLUSIONS It is suggested that less than 125 μ M of TG could protect the cells from H2O2 induced cell damage by down regulating the caspases and up regulating the Bcl-2 and Bcl-XL expression. Therefore, we suggest the trigonelline could be useful for treatment of oxidative stress mediated cardiovascular diseases in future.

Positive regulations of adipogenesis by Italian ryegrass [Lolium multiflorum] in 3T3-L1 cells.

Back groundIntramuscular fat deposition in the meat animal is relatively new strategy for developing the meat quality. Fat deposition is largely depending on the adipocyte proliferation and differentiation. Therefore, we investigated the effect of chloroform extract of L. multiflorum [CELM] on cell proliferation, lipid accumulation and adipocyte differentiation in 3T3-L1 cells and body weight of mouse.ResultsWe identified 6,9-Octadecatrienoic acid, Hexadecanoic acid, 2-hydroxypropanoic acid, butane-2,3-diol and hexane-1,2,3,4,5,6-hexaol in CELM. L. multiflorum extract increased the cell viability, lipid accumulation, cell cycle progression and key transcriptional and secretory factors like PPRAγ2, C/CEBP-α, adiponectin, aP2, GLUT-4, FAS and SREBP-1 mRNA expression as compared with control cells. For in-vivo, mice administered with CELM significantly increased body weight throughout the experiment periods. Further, the identified fatty acids like 3, 6, 9-Octadecatrienoic acid and Hexadecanoic acid was docked with target protein [PPRAγ2] using HEX 6.12. The least binding energy considered as high affinity with target protein. The maximum affinity with the target protein was observed in the Hexadecanoic acid followed by 3, 6, 9-Octadecatrienoic acid. The binding efficacy of Hexadecanoic acid and 3, 6, 9-Octadecatrienoic acid to the active site of PPAR-γ2 may be enhanced the adipocyte differentiations.ConclusionThese findings suggest that CELM stimulates adipogenesis via activating the PPARγ-mediated signaling pathway in adipocyte which could be useful for the development of meat quality in animals.

The effect of lactation number, stage, length, and milking frequency on milk yield in Korean Holstein dairy cows using automatic milking system

Objective The aim of the current study was to describe the relationship between milk yield and lactation number, stage, length and milking frequency in Korean Holstein dairy cows using an automatic milking system (AMS). Methods The original data set consisted of observations from April to October 2016 of 780 Holstein cows, with a total of 10,751 milkings. Each time a cow was milked by an AMS during the 24 h, the AMS management system recorded identification numbers of the AMS unit, the cow being milking, date and time of the milking, and milk yield (kg) as measured by the milk meters installed on each AMS unit, date and time of the lactation, lactation stage, milking frequency (NoM). Lactation stage is defined as the number of days milking per cows per lactation. Milk yield was calculated per udder quarter in the AMS and was added to 1 record per cow and trait for each milking. Milking frequency was measured the number of milkings per cow per 24 hour. Results From the study results, a significant relationship was found between the milk yield and lactation number (p<0.001), with the maximum milk yield occurring in the third lactation cows. We recorded the highest milk yield, in a greater lactation length period of early stage (55 to 90 days) at a 4× milking frequency/d, and the lowest milk yield was observed in the later stage (>201 days) of cows. Also, milking frequency had a significant influence on milk yield (p<0.001) in Korean Holstein cows using AMS. Conclusion Detailed knowledge of these factors such as lactation number, stage, length, and milking frequency associated with increasing milk yield using AMS will help guide future recommendations to producers for maximizing milk yield in Korean Dairy industries.

Potential role of marine algae extract on 3T3-L1 cell proliferation and differentiation: an in vitro approach

BackgroundFrom ancient times, marine algae have emerged as alternative medicine and foods, contains the rich source of natural products like proteins, vitamins, and secondary metabolites, especially Chlorella vulgaris (C. vulgaris) contains numerous anti-inflammatory, antioxidants and wound healing substances. Type 2 diabetes mellitus is closely associated with adipogenesis and their factors. Hence, we aimed to investigate the chemical constituents and adipogenic modulatory properties of C. vulgaris in 3T3-L1 pre-adipocytes.ResultsWe analysed chemical constituents in ethanolic extract of C. vulgaris (EECV) by LC–MS. Results revealed that the EECV contains few triterpenoids and saponin compounds. Further, the effect of EECV on lipid accumulation along with genes and proteins expressions which are associated with adipogenesis and lipogenesis were evaluated using oil red O staining, qPCR and western blot techniques. The data indicated that that EECV treatment increased differentiation and lipid accumulation in 3T3-L1 cells, which indicates positive regulation of adipogenic and lipogenic activity. These increases were associated with up-regulation of PPAR-γ2, C/EBP-α, adiponectin, FAS, and leptin mRNA and protein expressions. Also, EECV treatments increased the concentration of glycerol releases as compared with control cells. Troglitazone is a PPAR-γ agonist that stimulates the PPAR-γ2, adiponectin, and GLUT-4 expressions. Similarly, EECV treatments significantly upregulated PPAR-γ2, adiponectin, GLUT-4 expressions and glucose utilization. Further, EECV treatment decreased AMPK-α expression as compared with control and metformin treated cells.ConclusionThe present research findings confirmed that the EECV effectively modulates the lipid accumulation and differentiation in 3T3-L1 cells through AMPK-α mediated signalling pathway.

Enhancement of Nutritional Quality of Italian Ryegrass Mediated Silage by Supplemented with Lactic Acid Bacteria and Chlorella

The aim of present study was to improve the quality of silage using lactic acid bacteria (LAB) and chlorella as a supplement. Italian ryegrass (IRG) mediated silage was prepared with lactic acid bacteria (L. plantarum) and different concentration of chlorella. We analyzed the nutritional profiles such as crude protein (CP), acid detergent fiber (ADF) neutral detergent fiber (NDF), total digestible nutrient (TDN) and in-vitro dry matter digestibility (IVDMD), microbial counts and fermentative acids such as lactic acid, acetic acid and butyric acid in the control and experimental silage after three months. It shows increased crude protein content and also maintains the rest of nutritional values as compared with control silage. LAB inoculation with chlorella as supplementation slightly reduced the pH of the silage. In addition, it increased the fermentative acids production as compared with control silage and inhibits the undesired microbial growth especially fungi in the silage. Therefore, we suggest that LAB inoculation and chlorella supplementation to the IRG mediated silage could be improved the nutritional quality of the silage which is an intrinsic feature for the application in the preparation of animal feeds and functional foods.

Addition of Novel Lactobacillus plantarum KCC-10 and KCC-19 to Improve Fermentation Quality and Characterization of Italian Ryegrass Silage

ABSTRACT An investigation was carried out to evaluate the potential role of Lactobacillus Plantarum KCC-10 and KCC-19 on the quality and fermentation characterization of Italian ryegrass (IRG) silages. The physiochemical properties of IRG silage such as crude protein content, neutral detergent fiber, acid detergent fiber, total digestible nutrient and in vitro dry matter digestibility were not affected by KCC-10 and KCC-19. The pH of IRG silage in KCC-10 and KCC-19 treatments decreased compared to the control (p<0.05), while the lactic acid content in KCC-10 and KCC-19 treatments increased compared to the control (p<0.05). In addition, the number of lactic acid bacteria (LAB) in the KCC-10 treatment increased compared to the control (p<0.05). The number of lactic acid bacteria in KCC-19 increased, but there was no significant difference in all treatments. Therefore, we recommend L. plantarum KCC-10 and KCC-19 as potential additive candidates in IRG silage with lots of advantages.(