Morphological and Molecular Characterization of Lactic acid Bacteria from intestine of Freshwater Fishes and Their Possible Role in Probiotic Feeding in Aquaculture

2015 ◽  
Vol 5 (3) ◽  
pp. 101-106
Author(s):  
Dinesh Kumar Kumawat ◽  
Nadim Chishty
Keyword(s):  
Antibacterial Activity ◽  
Lactic Acid ◽  
Salt Tolerance ◽  
Lactic Acid Bacteria ◽  
Bile Salt ◽  
Micrococcus Luteus ◽  
Food Preservation ◽  
Rrna Gene ◽  
Gram Staining

  Lactic acid bacteria (LAB) are a group of gram positive, non spore forming, cocci or rod shaped, catalase negative organisms. Generally they are known as "GRAS" (Generally Recognized As Safe) organisms. In the present study, an attempt was made to isolate lactic acid bacteria from gut of two freshwa-ter fishes in Udaipur region to obtain various efficient LAB strains for indus-trial applications. A total of 12 bacterial isolates were isolated from different fishes caught from Pichola lake, Udaipur on MRS agar at 37oC. All the isolates were tested for catalase activity, colony morphology and gram staining. Further these isolates were checked for technological properties (antibacterial activity, antibiotic susceptibility and bile salt tolerance). Based on technologi-cal properties results, two isolate (F-4 & F-11) have been tentatively selected for use as probiotic cultures in the aquaculture industries. They have strong antibacterial activity against various pathogens viz. Staphylococcus aureus (ATCC 9144), Micrococcus luteus (ATCC 10240), Listeria monocytogens (MTCC 1143) and have bile salt tolerance limit up to 0.7%. These are the main features for selection of probiotic candidates. To differentiate the same species molecular typing was attempted, 16S rRNA gene amplification was done using specific 16S rDNA primers and amplified gene products were di-gested with different restriction endonuclease enzymes viz EcoRI and HindIII. A total of 2 different groups out of 12 isolates were obtained. This finding reveals that isolated LAB are useful for aquaculture and also used in food preservation.

Selection and characterization of potential probiotic Lactobacilli spp isolated from chicken feces may be used as a potent antibacterial agent

2016 ◽  
Vol 35 (1) ◽  
Author(s):  
Nazila Aazami ◽  
Enayatollah Kalantar ◽  
Helen Poormazaheri ◽  
Nassimeh Setayesh vali Pour ◽  
Gholamreza Salehi Jouzan
Keyword(s):  
Antibacterial Activity ◽  
Antibiotic Resistance ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Acid Tolerance ◽  
Resistance Test ◽  
Effect Of Temperature ◽  
Rdna Sequencing

<italic>Lactobacillus</italic> strains commonly used as probiotics which possess some properties such as resistance to gastric acidity and bile salts, production of antimicrobial compounds, the ability to modulate immune responses, and adhesion to gut tissues. This study aimed to isolate, identify, production and characterization of the probiotic strains. Total of 10 digestive tracts of chickens fed without antibiotics for presence of <italic>Lactobacillus spp</italic> was evaluated. All isolates were grown with different conditions like effect of temperature, pH, pepsine, bile, sodium chloride tolerance, aggregation, adhesion and antibiotic resistance test tested and then antibacterial activity was determined.In this study, 34 lactic acid bacteria were investigated as putative probiotic candidates. Based on the molecular identification using 16S rDNA sequencing the isolates belonged to <italic>Lactobacillus</italic> species. The strains showed potent antibacterial activity. The results on acid tolerance showed that all the strains tested survived at pH 2.0 to 5.5. <italic>L. agilis</italic> strain M18, <italic>L. salivarius</italic> strain M10, <italic>L.vaginalis</italic> strains M8 and M19, and <italic>L. crispatus</italic> strains Or2 and M10 showed the lowest sensitivity to pepsine. All the studied Lactobacillus strains were able to grow at both 15 and 45°C . Five Lactobacillus strains adhered well to the Caco-2 cells. The result of antibiotic resistance test of 34 lactic acid bacteria showed that most of them were resistant or intermediate. Based on 34 <italic>Lactobacillus</italic> strains, <italic>L. vaginalis</italic> strain M8 and <italic>L. reuteri</italic> strain M6 were selected for the further in vivo assays and possible eventual use as prophylactic and therapeutic agent.

scholarly journals Lactic Acid Bacteria Isolated from Locally Produced Vinegars and Their Antibacterial Activity Against Foodborne Bacteria

2019 ◽  
Vol 1 (2) ◽  
pp. 1-7
Author(s):  
Agnes Lee Chiu Nee ◽  
Mohd Nizam Lani ◽  
Rozila Alias ◽  
Zaiton Hassan
Keyword(s):  
Antibacterial Activity ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
Titratable Acidity ◽  
Diffusion Method ◽  
Phenotypic Characterization ◽  
Black Rice ◽  
Gram Staining ◽  
Rice Vinegar

Vinegars are most widely used as preservatives in food industry. Vinegars are known for their health benefits; however, the roles of vinegar-associated microflora in locally produced vinegars are not well established. The objectives of this study are to isolate and identify the lactic acid bacteria (LAB) from black rice vinegar and coconut vinegar, measure their pH and titratable acidity, and determine their antibacterial activity. LAB was isolated using cultural method. Phenotypic characterization of LAB was carried out using Gram-staining, oxidase test, catalase test and API 50 CHL Kit. Results from API 50 CHL Kit confirmed that BRV03M strain from black rice vinegar and CV03M strain from coconut vinegar were Lactobacillus paracasei ssp. paracasei. The identified bacteria in both samples were consistent as L. paracasei using 16S rDNA gene sequences with 93% and 99% similarity, respectively. The pH and titratable acidity percentage of both vinegars were also determined. The stability of Cell Free Supernatant-Lactic Acid Bacteria (CFS-LAB) strains within 14 days on their inhibition against selected pathogenic bacteria was determined using agar well diffusion method. The CFS-LAB strain isolated from black rice vinegar (BRV03M) was more stable within 14 days than coconut vinegar in inhibiting tested bacteria, suggesting this strain has great potential as natural antibacterial agents.

scholarly journals Evaluation of the probiotic and postbiotic potential of lactic acid bacteria from artisanal dairy products against pathogens

2021 ◽  
Vol 15 (01) ◽  
pp. 102-112
Author(s):  
Nazar Hussain ◽  
Muhammad Tariq ◽  
Per Erik Joakim Saris ◽  
Arsalan Zaidi
Keyword(s):  
Antibacterial Activity ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
16S Rrna Gene Sequencing ◽  
Lactobacillus Delbrueckii ◽  
Human Consumption ◽  
Rrna Gene ◽  
Microdilution Method

Introduction: Probiotic and postbiotic potential of thirty-two strains of lactic acid bacteria (LAB), obtained earlier from artisanal dairy sources in Pakistan, have been investigated against major multi-drug resistant (MDR) and food borne pathogenic bacteria. Methodology: LAB strains were identified by 16S rRNA gene sequencing and their antibacterial activity was assessed by the microdilution method. Four LAB isolates, Weissella confusa PL6, Enterococcus faecium PL7, and Lactobacillus delbrueckii PL11 and PL13 were shortlisted. Their ability to degrade lactose and safety for human consumption in terms of hemolysis and antibiotic susceptibility were assessed in vitro. The antibacterial components in the cell-free supernatants (CFSs) of isolate cultures were characterized biochemically by HPLC. Results: Acid neutralization but not protease treatment abolished the antibacterial activity of CFSs. Lactic, acetic and propionic acids were the main acids in the CFSs, and acid production peaked in the stationary phase of growth. The antibacterial activity of the LAB cultures resulted from secretion of organic acids that lowered the pH. The strains exhibited variable ability to degrade lactose and were non-hemolytic and susceptible to the most common antibiotics. Conclusions: These LAB strains are probiotic candidates for further investigation of their postbiotic role in naturally preserving processed foods and for attenuation of lactose intolerance.

scholarly journals Homofermentative lactic acid bacteria of a traditional cheese, Comlek peyniri from Cappadocia region

2005 ◽  
Vol 72 (1) ◽  
pp. 19-24 ◽  
Author(s):  
Cisem Bulut ◽  
Hatice Gunes ◽  
Burcu Okuklu ◽  
Sebnem Harsa ◽  
Sevda Kilic ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Molecular Identification ◽  
Lactic Acid Production ◽  
Skim Milk ◽  
Gas Production ◽  
Central Anatolia ◽  
Rrna Gene ◽  
Ph Change ◽  
Gram Staining

Comlek peyniri is a typical artisanal cheese in Central Anatolia. This type of cheese was made by using the indigenous lactic acid bacteria (LAB) flora of cow or ewes' milk. Majority of the samples were taken from fresh cheese because the aim was to isolate homofermentative LAB. Initially 661 microbial isolates were obtained from 17 cheese samples. Only 107 were found to be homofermentative LAB. These isolates were selected and identified by using both phenotypic and molecular methods. Phenotypic identification included curd formation from skim milk, catalase test, Gram staining and light microscopy, growth at different temperatures and salt concentrations, arginine hydrolysis, gas production from glucose, and carbohydrate fermentation. Molecular identification was based on the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the 16S rRNA gene-ITS (internally transcribed spacer) region. By combining the phenotypic and molecular identification results, isolates belonging to each of the following genera were determined at species or subspecies level: 54 Lactococcus lactis subsp. lactis, 21 Enterococcus faecium, 3 Ec. faecalis, 2 Ec. durans, 10 Ec. sp., 15 Lactobacillus paracasei subsp. paracasei, and 2 Lb. casei strains. Technological characterisation was also performed by culturing each of the strains in UHT skim milk, and by monitoring pH change and lactic acid production at certain time intervals through the 24 h incubation. Results of the technological characterisation indicated that 33% of the isolates (35 strains) were capable of lowering the pH of UHT milk below 5·3 after 6 h incubation at 30 °C. Thirty four of these strains were Lc. lactis subsp. lactis, and only one was an Ec. faecium strain.

scholarly journals Lactic Acid Bacteria Diversity and Characterization of Probiotic Candidates in Fermented Meats

Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1519
Author(s):  
Elvina Parlindungan ◽  
Gabriele A. Lugli ◽  
Marco Ventura ◽  
Douwe van Sinderen ◽  
Jennifer Mahony
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Organic Acids ◽  
Bile Salt ◽  
Antibiotic Susceptibility ◽  
Dominant Species ◽  
Fermented Foods ◽  
Probiotic Potential ◽  
Excellent Source

Probiotics are defined as live microorganisms which confer health benefits to the host when administered in adequate amounts. Many lactic acid bacteria (LAB) strains have been classified as probiotics and fermented foods are an excellent source of such LAB. In this study, novel probiotic candidates from two fermented meats (pancetta and prosciutto) were isolated and characterized. LAB populations present in pancetta and prosciutto were evaluated and Lactiplantibacillus plantarum was found to be the dominant species. The antagonistic ability of selected isolates against LAB and non-LAB strains was investigated, in particular, the ability to produce anti-microbial compounds including organic acids and bacteriocins. Probiotic characteristics including antibiotic susceptibility, hydrophobicity and autoaggregation capacity; and ability to withstand simulated gastric juice, bile salt, phenol and NaCl were assessed. Among the characterized strains, L. plantarum 41G isolated from prosciutto was identified as the most robust probiotic candidate compared. Results from this study demonstrate that artisanal fermented meat is a rich source of novel strains with probiotic potential.

scholarly journals Exploration of the Microbial Biodiversity Associated with North Apulian Sourdoughs and the Effect of the Increasing Number of Inoculated Lactic Acid Bacteria Strains on the Biocontrol against Fungal Spoilage

Fermentation ◽  
2019 ◽  
Vol 5 (4) ◽  
pp. 97 ◽  
Author(s):  
Mattia Pia Arena ◽  
Pasquale Russo ◽  
Giuseppe Spano ◽  
Vittorio Capozzi
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Shelf Life ◽  
Restriction Analysis ◽  
Rflp Analysis ◽  
Food Preservation ◽  
Rrna Gene ◽  
Microbiological Analysis ◽  
Bread Making ◽  
Dna Restriction

In this study, we explored the diversity of yeasts and lactic acid bacteria (LAB) associated with six spontaneous sourdough fermentations from the northern part of the Apulian region (Italy). Bacterial and yeast isolates from sourdough were investigated by amplified ribosomal DNA restriction analysis (ARDRA) and restriction fragment length polymorphism (RFLP) analysis, respectively. The identification of the isolates was confirmed by sequencing bacterial 16S gene and yeast ITS1–5.8S–ITS2 rRNA gene amplicons. Microbiological analysis of all sourdough samples revealed that LAB and yeast counts ranged between 1.7 × 105 and 6.5 × 108 cfu/g, and 7.7 × 105 and 2.5 × 107 cfu/g, respectively. The molecular identification at species level revealed the occurrence of Lactobacillus plantarum as the dominant LAB and Saccharomyces cerevisiae as the dominant yeast species in all different sourdough samples. Then, the ability of all isolated strains to inhibit and/or reduce the growth of several selected fungi was valued through the overlay method. In light of their antifungal performances, ten LAB strains were inoculated, singularly and in combination, in subsequent bread-making trials. Overall, we confirmed the potential of LAB to extend the shelf life of bread through spoilage inhibition and, for the first time, we observed a synergistic effect due to the combination of several isolated LAB on the inhibition behavior against selected fungal spoilage strains. Our findings suggest the exploration of a LAB-based approach in order to extend the shelf life of bread, reducing, at the same time, the use of chemical agents for food preservation.

scholarly journals Purification and Characterization of a Novel Bacteriocin Pediocin Z-1 Produced by Pediococcus Pentosaceus Z-1 Screened From Jinhua Ham

2020 ◽  
Author(s):  
Xiaohong Yu ◽  
Zhicai Yang ◽  
Keyue Li ◽  
Rui Liu ◽  
Huijie Pei ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Antibacterial Activity ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Food Industry ◽  
Acid Base ◽  
Pediococcus Pentosaceus ◽  
Base Stability ◽  
Jinhua Ham

Abstract Background: Lactic acid bacteria (LAB) can produce bacteriostatic substances, among which bacteriocins attracted wide attention in food preservation for decades. Up to date, nisin (class I bacteriocins) has been considered to be the only bacteriocin produced by Lactococcus lactis strains for commercial use. Moreover, there are many other reports concerning the isolated bacteriocins for potential application in food industry while some exhibited a narrow bactericidal spectrum, thermal stability and low acid-base stability. Jinhua ham is a representative of traditional dry-cured meat product in China. The microbial community structure and diversity was speculated to be responsible for flavor and quality formation of Jinhua ham, protecting from the spoilage microorganisms contamination. However, no studies was performed to investigate the bacteriocin from LAB in Jinhua ham. Thus, the objective of this study was to screen out high-efficiency, safe and non-toxic bacteriocin-producing lactic acid bacteria (LAB) from Jinhua Ham and subsequently perform the purification, identification and characterization of its bacteriocin. Results: The bacteriocin-producing LAB was screened from Jinhua ham and then designated as Pediococcus pentosaceus Z-1 by colony morphology and 16S rDNA sequencing. The bacteriocin was then crudely extracted from the bacterial cell-free supernatant by pH adsorption, and further purified by cellulose DEAE-52 ion exchange and Sephadex G-50 chromatography columns. The tricine-SDS-PAGE electrophoresis showed a highly purified protein band with 8227.35 Da with 60 amino acids identified by MALDI-TOF-MS analysis. The bacteriocin was named as pediocin Z-1 and its antibacterial activity exhibited an acid-base stability between pH 2-10 and a thermal stability at a range of 50-110°C. The pediocin Z-1 was sensitive to proteases and showed an inhibitory effect against Gram-positive bacteria and Gram-negative bacteria, including Listeria monocytogenes, Staphylococcus aureus, Salmonella typhimurium, Salmonella potsdam and Escherichia coli. Conclusions: Our data suggest that pediocin Z-1 has a comparable effect of antibacterial activity with nisin and it is promising that pediocin Z-1 has great potential for application prospect in the food industry.

scholarly journals Isolation of Lactic Acid Bacteria from Idli Batter and Screening of Characters as Potential Starter Cultures for Development of Functional Food Using Bacopa monnieri

2020 ◽  
Vol 2 (1) ◽  
pp. 82
Keyword(s):  
Lactic Acid ◽  
Salt Tolerance ◽  
Lactic Acid Bacteria ◽  
Bile Salt ◽  
Starter Culture ◽  
Antibacterial Properties ◽  
Bacopa Monnieri ◽  
Growth Patterns ◽  
Heat Sensitivity ◽  
Idli Batter

Functional foods are whole, fortified, enriched, or enhanced foods that provide health benefits to humans regularly. Numerous plant foods or physiologically active ingredients derived from plants have been investigated for their role in disease prevention and health. Natural lactic acid fermentation of Idli batter by lactic acid bacteria and yeast makes Indian breakfast as a potential source of probiotic microorganisms. Briefly, the organism was isolated; the predominant isolate was considered on the basis of colony morphology, staining techniques, and biochemical tests. Growth conditions of the organism, such as pH, salt tolerance, bile salt tolerance, heat sensitivity, were studied. Bacopa monnieri (Brahmi juice) was extracted and fermented with predominant isolate as a starter culture. Organoleptic, antioxidant, and antibacterial properties of the fermented product were evaluated. On staining, the predominant isolate was identified as Gram-positive non-spore-forming, non-motile rod shape bacteria. The phenotypic characteristics were further resolved on the basis of growth patterns in pH, salt, bile salt, and heat sensitivity. In conclusion, the study of biological efficacies of fermented extracts of Bacopa monnieri reveals the potent activities in the field of science.

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