The screening of probiotic lactic acid bacteria from honey of stingless bee from West Sumatra, Indonesia and using as starter culture (original) (raw)

Probiotic Properties of Bacillus Strains Isolated from Stingless Bee (Heterotrigona itama) Honey Collected across Malaysia

International Journal of Environmental Research and Public Health, 2019

This study aimed to isolate, identify, and evaluate the probiotic properties of Bacillus species from honey of the stingless bee Heterotrigona itama. Bacillus spp. were isolated from five different H. itama meliponicultures, and the isolates were characterized through Gram-staining and a catalase test. Tolerance to acidic conditions and bile salt (0.3%), hydrophobicity, and autoaggregation tests were performed to assess the probiotic properties of the selected isolates, B. amyloliquefaciens HTI-19 and B. subtilis HTI-23. Both Bacillus isolates exhibited excellent antimicrobial activity against both Gram-positive and Gram-negative bacteria and possessed significantly high survival rates in 0.3% bile solution for 3 h. Their survival rates in acidic conditions were also comparable to a commercial probiotic strain, Lactobacillus rhamnosus GG. Interestingly, the hydrophobicity and autoaggregation percentage showed no significant difference from L. rhamnosus GG, a commercial probiotic str...

Probiotic potency of Lactobacillus plantarum KX519413 and KX519414 isolated from honey bee gut

FEMS microbiology letters, 2018

The Indian honey bee Apis cerana indica, which harbors an abundant and diverse range of lactic acid bacteria (LAB) in their gut with beneficial effects, was used as the source for the isolation of LAB. In the present study, two LAB isolates from honey bee gut were selected primarily based on their phenotypic and selective biochemical characterization, followed by PCR and identified using 16S rRNA sequencing as Lactobacillus plantarum and were registered in National Centre for Biotechnology Information under accession number KX519413 and KX519414. The probiotic potency of test strains indicated their survivability at acidic pH, bile salts and viability in simulated gastric juice enabling them to withstand gastrointestinal tract conditions. Evaluation of cell surface properties suggested that they possess an important defense mechanism against the pathogen since they are hydrophobic, auto-aggregative and have co-aggregative ability. Further, efficient exopolysaccharide production by t...

Characterization of probiotic properties and antimicrobial activity of lactic acid bacteria from stingless bee gut

International journal of health sciences

Lactic acid bacteria (LAB) are known as the major group of probiotics that possess different beneficial properties. Several studies had reported the isolation of LAB from stingless bees. However, the isolation of LAB specifically from stingless bee gut is still limited. The bee microbiome frequently hosts LAB which demonstrated beneficial effects such as reducing bacterial and parasite infections as well as increasing honey output in beehives. This study aimed to characterize the probiotic properties and antimicrobial activity of LAB from stingless bee gut. A total of five LAB strains were successfully isolated from three different species of stingless bee known as Heterotrigona itama (HIT), Geniotrigona thoracica (GTH), and Tetragonula laeviceps (TLA). All LAB isolates were assessed for in vitro probiotic activity such as acid tolerance, bile salt tolerance, tolerance to simulated gastric juice, surface hydrophobicity, and also their antimicrobial activity. The percentage viability...

Isolation and characterization of lactic acid bacteria from Sabah (North Borneo) stingless bees for probiotic and food applications

International Journal of Food Properties

Stingless bees harbor lactic acid bacteria (LAB) which possesses multiple beneficial properties. However, there is no report on LAB in stingless bee from Sabah and their products. This study aimed to isolate LAB from several stingless bee species and their products as well as to perform characterization and safety assessments. A total of 104 strains were isolated and seven potential antimicrobial LAB isolates were identified from stingless bee and their products. Characterization, identification, and assessments were performed on seven (A2b, B3b, P1b, H4b, A6, B5, and B10) LAB that exert potential antimicrobial activity against Listeria monocytogenes ATCC 7644. The A6 isolate was closely related to the Lactobacillus pentosus species whereas isolates B5 and B10 were closely related to Weissella paramesenteroides species. Finally, isolates P1b, H4b, B3b, and A2b were closely related to the Enterococcus sp. These seven LAB isolates were able to survive in stimulated gastrointestinal tract conditions (acidic, salt, bile salt, and temperature). The carbon fermentation, proteolytic activities, acidification, milk coagulation, and wide antibiotic susceptibility testing of seven LAB isolates revealed their potential used as a probiotic and fermentation purposes. Therefore, more studies are warranted to investigate the potential of these LAB isolates towards applications in probiotic and fermentation.

Identification and Antimicrobial Activity of Lactic Acid Bacteria Isolated from Tualang Bee Honey

Malaysian Journal of Fundamental and Applied Sciences, 2023

Malaysia multifloral Tualang honey usually collected from the combs of Asia rock bees (Apis dorsata), which build their hives high up in the Tualang tree (Koompassia excelsa). Tualang honey is used commonly as a medicinal product and as food in Malaysia. lactic acid bacteria (LAB) strains have been discovered to exhibit antimicrobial properties especially in Malaysia. However, the research on LAB found from honey are still scarce. The study was conducted to isolate and identify lactic acid bacteria (LAB) from Tualang honey bee from East Coast Peninsular Malaysia. The Tualang honey with LAB strains were further tested against several pathogenic bacteria by using well-diffusion method. The LAB were primarily identified by colony morphology, microscopy of Gram`s stain, biochemical tests and 16s rRNA sequencing method. The 10 LAB strains that has been identified as Leuconostoc mesenteroides (5 strains), Lactobacillus kunkeei (4 strains), and Lactobacillus farraginis (1 strains) were selected for the characterization of antimicrobial agents produced by LAB. The findings shows that 5 strains exhibited strong antimicrobial activity against Staphylococcus epidermidis and Pseudomonas aeruginosa results in excellent inhibition zones diameters larger than 15 mm. This study indicated that LAB isolated from Tualang honey has potential antimicrobial activity against pathogenic bacteria and can be further characterized for health benefits and potential use in food industry at Malaysia.

Lactic Acid Bacteria from Honey Bees Digestive Tract and Their Potential as Probiotics

Proceedings of the International Conference and the 10th Congress of the Entomological Society of Indonesia (ICCESI 2019), 2020

Honey bees (Apis spp.) are social insects that possess unique gut microbiota community. There are three main phyla that dominate the honey bees gut microbiota; Proteobacteria, Firmicutes, and Actinobacteria in which several species among them belong to the lactic acid bacteria (LAB). In general, some species of LAB can be developed as potential probiotics due to their capability to produce bioactive compounds such as organic acid (lactic acid, acetate acid and formic acid), hydrogen peroxide (H 2 O 2), ethanol, enzymes, benzoate, antimicrobial peptides (AMPs), free fatty acid, and volatile compounds that in a synergistic action work as broad-spectrum antimicrobials toward several pathogens. Several strains of LAB that living in symbiosis within the digestive tract of honey bees have been isolated, among them are quite promising to be developed as probiotics. Multiple studies have shown the inhibitory effect of LAB from honey bees against pathogenic microorganisms. Formic acid and lactic acid produced by symbiotic LAB could decrease the environmental pH of wounds, therefore prevent the growth of pathogenic microbes. Furthermore, the volatile compound produced by LAB is toxic, while H 2 O 2 in small numbers is needed for optimal wound recovery. LAB capable of producing broad-spectrum antibacterial compounds such as Serratia marcescens, Eschericia coli, methicillin-resistant Staphylococcus aureus, Klebsiella aerogenes, Salmonella typhi, Pseudomonas spp., Klebsiella spp., Proteus spp., S. aureus, and Bacillus subtilis. Additionally, LAB possess antioxidant activity and to adhere the epithelial cells. This paper aimed to discuss varieties of LAB from the digestive tract of honey bees and their potential as probiotics that could benefit healthcare.

Probiotic capability of novel lactic acid bacteria isolated from worker honey bees gut microbiota

FEMS Microbiology Letters, 2021

The study aimed to evaluate the probiotic and safety properties of lactic acid bacterial (LAB) strains isolated from the gut microbiota of honey bee Apis mellifera L., since this source remains a promising reservoir of microbial diversity. A total of five bacterial isolates were molecularly identified using 16S rRNA gene sequencing as Enterococcus faecalis-HBE1, Lactobacillus brevis-HBE2, Enterococcus faecalis-HBE3, Enterococcus faecalis-HBE4 and Lactobacillus casei-HBE5. Gut tolerance conditions (low pH and bile salt) were evaluated. Exopolysaccharides (EPS) production, hemolytic, antioxidant activity, resistance toward antibiotics and technological characteristics (starter activity, pH and proteolysis) were examined. The five isolates showed a high survival rate (>95%), under gastrointestinal tract conditions indicating excellent potential for application as probiotics. The isolates showed no hemolytic activities and good acidification rates in the range of pH 4.6–4.98 after in...

Screening and Characterization of Putative Probiotic Lactobacillus Strains from Honey Bee Gut (Apis mellifera)

Journal of Advances in Microbiology, 2018

The objective of this work was to isolate, identify and characterize lactobacilli strains from the intestinal tract of honey bees as putative probiotics. We obtained eighty-five isolates. At the end of screening based on physiological properties, 17 isolates were pre-selected and their resistance to gastrointestinal stress was evaluated. Twelve (12) with good resistance after 3 h exposure to low pH values (pH2, pH3) were subjected to determination of their in vitro BSH activity. The research of the bsh-A, bsh-B, Bsh1 and Bsh-Lp1 genes encoding the BSH enzyme was also conducted. Four isolates (H46, H82, H21 and H28) were resistant, seven others tolerant (H6, H15, H47, H24, H67, H44, H80) and only one was sensitive (H63) to oxgall bile salt. Determination of BSH activity revealed that all strains hydrolyze bile salts, with a preference for oxgall as opposed to Original Research Article

ANTIBIOTIC SUSCEPTIBILITY, ANTIBACTERIAL ACTIVITY AND PROBIOTIC CHARACTERISATION OF ISOLATED Lactobacillus brevis STRAINS FROM Heterotrigona itama HONEY

Heterotrigona itama (Family: Apidae, Tribe: Meliponini, Genus: Trigona) is one of the stingless bee in Meliponiculture in Malaysia. H. itama honey is reported to be a good reservoir for novel lactic acid bacteria (LAB) with probiotic properties. In this study, five Lactobacillus brevis strains (strain Ibr-42, strain 37901, strain ATCC 367, strain NJ42 and strain KLDS) were previously isolated and identified from H. itama honey obtained from local stingless beekeepers in the coastal areas in Kelantan and Terengganu, were evaluated for antibiotic resistance, antibacterial activity, resistance to low pH, tolerance to bile salts and haemolytic activity. The results indicated that all five strains of L. brevis were susceptible to chloramphenicol, ampicillin, erythromycin, and tetramycin, but resistant to kanamycin. In terms of antagonistic activity among L. brevis, it was found antagonistic activity was minimum. For antibacterial activities of these strains against selected foodborne pathogenic bacteria by well diffusion method, L. brevis strain NJ42 exhibited the highest inhibition (24 mm) against Pseudomonas aeruginosa ATCC 15442. All Lactobacillus strains from H. itama honey were able to survive in pH 2 and 0.3% (w/v) bile salts concentration that mimic the conditions in the gastrointestinal system. The inability of L. brevis strains to exhibit β-haemolytic activity showing that haemolysis is not the virulence factor for these strains. These findings proved the isolated L. brevis in H. itama honey could be used as potential probiotic, envisaging its potential as one of functional foods for food industry.