Isolation and identification of Bifidobacterium species from human breast milk and infant feces in Indonesia

##plugins.themes.bootstrap3.article.main##

ROBIN DOSAN
SAMUEL OWEN MUDANA
CLARA MEIRINZHA PANG JULYANTO
EMILY TANIA PURNAMA
MARCELIA SUGATA
JUANDY JO
TJIE JAN TAN

Abstract

Abstract. Dosan R, Mudana SO, Julyanto CMP, Purnama ET, Sugata M, Jo J, Tan TJ. 2024. Isolation and identification of Bifidobacterium species from human breast milk and infant feces in Indonesia. Biodiversitas 25: 337-343. There has been a growing interest in identifying emerging probiotic strains because of their benefits for human health. Many bifidobacteria originated from humans have been reported to possess probiotics properties. They are commonly found in the intestine of breast-fed infants. Hence, this study aimed to isolate and identify bifidobacteria from human breast milk and infant fecal samples in Indonesia and evaluate their probiotic properties. Twenty colonies were isolated from two independent fecal samples and two independent breast milk samples. Ten isolates (BR1-M1, BR1-B1, BR2-5, BR2-6, BR2-12, BS2-PB3, BS2-PB5, BS2-PS1, BS2-PS2, BS2-MB1) showed a compatible phenotypic character with Bifidobacteria based on the Bergey’s Manual, including Gram-positive, irregular rods, no catalase activity, non-spore-forming, and non-motile. Subsequently, four isolates with similar carbohydrate fermentation patterns as Bifidobacterium spp. were selected for further molecular identification based on 16S rRNA gene sequencing analysis. The results showed that BR2-5 and BR2-6 were found to be closely related to Bifidobacterium animalis subspecies lactis with 100 and 98.39% similarity, respectively. Meanwhile, BS2-PS1 and BS2-PB3 were found to be closely related to Bifidobacterium breve with 100 and 98.26% similarity, respectively. Further investigation revealed that BR2-5 and BS2-PB3 were resistant to low pH (?4) and could tolerate the exposure of bile salts (1%). Both isolates survived under different oxidative stress conditions (aerobic and microaerophilic). In conclusion, BR2-5 and BS2-PB3 exhibited promising characteristics as probiotic candidates, though further investigations are required to substantiate these current findings.

##plugins.themes.bootstrap3.article.details##

References
City S, Sugata M, Tan TJ. 2011. Probiotic characterization of Bacillus subtilis SM10.1. J Phys: Conf Ser 1918: 052025. DOI:10.1088/1742-6596/1918/5/052025.
Chichlowski M et al. 2020. Bifidobacterium longum Subspecies infantis ( B. infantis) in Pediatric Nutrition: Current State of Knowledge. Nutrients 12(6): 1581. DOI: 10.3390/NU12061581.
Eglash A, Simon L. 2017. ABM Clinical Protocol #8: Human Milk Storage Information for Home Use for Full-Term Infants, Revised 2017. Breastfeed Med?: Offl J Acad Breastfeed Med 12(7):.390–395. DOI: 10.1089/BFM.2017.29047.AJE.
Fuochi V et al. 2015. Evaluation of resistance to low pH and bile salts of human Lactobacillus spp. isolates. Int J Immunopathol Pharmacol 28(3): 426–433. DOI: 10.1177/0394632015590948.
Goodfellow M. 2012. Phylum XXVI. Actinobacteria phyl. nov. In: Michael Goodfellow (eds.). Bergey’s Manual® of Systematic Bacteriology. Springer, New York.
Guo Q et al. 2017. The NAD+-dependent deacetylase, Bifidobacterium longum Sir2 in response to oxidative stress by deacetylating SigH (?H) and FOXO3a in Bifidobacterium longum and HEK293T cell respectively. Free Radic Biol Med 108: 929-939. DOI: 10.1016/j.freeradbiomed.2017.05.012.
Hanidah II et al. 2019. Characterization of Probiotic Bacterial Candidates from Jatinangor-Indonesia Breast Milk. Int J Adv Sci Eng Inf Technol 9(5): 1649–1655. DOI: 10.18517/ijaseit.9.5.10124.
Hedberg M et al. 2008. Sugar fermentation in probiotic bacteria – an in vitro study. Oral Microbiol Immunol 23(6): 482–485. DOI: 10.1111/J.1399-302X.2008.00457.X.
Junick J, Blaut M. 2012. Quantification of human fecal Bifidobacterium species by use of quantitative real-time PCR analysis targeting the groEL gene. Appl Environ Microbiol 78(8): 2613–2622. DOI: 10.1128/aem.07749-11.
Kusharyati DF et al. 2020. Isolation of Bifidobacterium from Infant’s Feces and Its Antimicrobial Activity. Digit Press Life Sci 2: 00002. DOI: 10.29037/DIGITALPRESS.22326.
Li G. 2012. Intestinal Probiotics: Interactions with Bile Salts and Reduction of Cholesterol. Proced Environ Sci 12(B): 1180–1186. doi: 10.1016/J.PROENV.2012.01.405.
Matsuki T et al. 1999. Distribution of bifidobacterial species in human intestinal microflora examined with 16S rRNA-gene-targeted species-specific primers. Appl Environ Microbiol 65(10): 4506–4512. DOI: 10.1128/aem.65.10.4506-4512.1999.
Mattarelli P et al. 2017. The Bifidobacteria and Related Organisms?: Biology, Taxonomy, Applications.Massachusetts, Academic Press.
Modesto M. 2018. Isolation, Cultivation, and Storage of Bifidobacteria. In The Bifidobacteria and Related Organisms: Biology, Taxonomy, Applications. Massachusetts, Academic Press.
Parlindungan E et al. 2021. Lactic acid bacteria diversity and characterization of probiotic candidates in fermented meats. Foods 10(7): 1519. DOI: 10.3390/foods10071519.
Penders J et al. 2006. Factors influencing the composition of the intestinal microbiota in early infancy. Pediatrics 118(2): 511–521. DOI: 10.1542/PEDS.2005-2824.
Picard C et al. 2005. Review article: bifidobacteria as probiotic agents – physiological effects and clinical benefits. Aliment Pharmacol Ther 22(6): 495–512. DOI: 10.1111/J.1365-2036.2005.02615.X.
Pokusaeva K, Fitzgerald GF, van Sinderen D. 2011. Carbohydrate metabolism in Bifidobacteria. Genes Nutr 6: 285–306. DOI: 10.1007/s12263-010-0206-6.
Sánchez B, De Los Reyes-Gavilán CG, Margolles A. 2006. The F1F0-ATPase of Bifidobacterium animalis is involved in bile tolerance. Environ Microbiol 8(10): 1825–1833. DOI: 10.1111/J.1462-2920.2006.01067.X.
Shah NP. 2011. Bacteria, Beneficial?Bifidobacterium spp.: morphology and physiology. Encycl Dairy Sci 381-387. DOI:10.1016/b978-0-12-374407-4.00043-1.
Shibata N, Toraya T. 2015. Molecular architectures and functions of radical enzymes and their (re)activating proteins. J Biochem 158(4): 271-292. DOI: 10.1093/jb/mvv078.
Shigwedha N, Jia L. 2013. Bifidobacterium in Human GI Tract: Screening, Isolation, Survival and Growth Kinetics in Simulated Gastrointestinal Conditions. Intechopen 281–308. DOI: 10.5772/50457.
Stackebrandt E, Goebel BM. 1994. Taxonomic note: A place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Intl J Syst Bacteriol 44(4): 846–849. DOI: 10.1099/00207713-44-4-846.
Vitellio P et al. 2019. Effects of Bifidobacterium longum and Lactobacillus rhamnosus on gut microbiota in patients with lactose intolerance and persisting functional gastrointestinal symptoms: A randomised, double-blind, cross-over study. Nutrients 11(4): 886. DOI: 10.3390/nu11040886.
Vlková E et al. 2004. Enumeration, isolation, and identification of bifidobacteria from infant feces. Fol Microbiol 49(2): 209–212. DOI: 10.1007/BF02931404.
Walter J. 2008. Ecological role of lactobacilli in the gastrointestinal tract: Implications for fundamental and biomedical research. Appl Environ Microbiol 74(16): 4985–4996. DOI: 10.1128%2FAEM.00753-08.
Watanabe M et al. 2012. Effect of respiration and manganese on oxidative stress resistance of Lactobacillus plantarum WCFS1. Microbiology 158(1): 293–300. DOI: 10.1099/MIC.0.051250-0.
Watson D et al. 2013. Selective carbohydrate utilization by lactobacilli and bifidobacteria. J Appl Microbiol 114(4): 1132–1146. DOI: 10.1111/JAM.12105.
World Health Organization. 2001. Probiotics in Food: Health and Nutritional Properties and Guidelines for Evaluation. Rome, Food and Agriculture Organization of the United Nations.
Zuo F et al. 2014. Combination of heterogeneous catalase and superoxide dismutase protects Bifidobacterium longum strain NCC2705 from oxidative stress. Appl Microbiol Biotechnol 98: 7523–7534. DOI: 10.1007/s00253-014-5851-z.