Journal of Food Science and Technology

, Volume 55, Issue 5, pp 1870–1879 | Cite as

Antimicrobial, antioxidant and probiotics characterization of dominant bacterial isolates from traditional fermented fish of Manipur, North-East India

  • Sambanduram Samarjit Singh
  • Surajit De Mandal
  • Esther Lalnunmawii
  • Nachimuthu Senthil Kumar
Original Article


Utonga-kupsu, Hentak and Ngari are traditional fermented fish products produced by the Manipuri people living in the North-Eastern part of India. The present study was designed with the aim to isolate, identify and characterize the microorganisms present in these fermented foods. Bacterial pure cultures were isolated using serially diluted samples and were further identified by conventional biochemical tests and Sanger sequencing of 16s rRNA gene. Results show that the number of bacterial count in Nutrient agar and Starch casein agar was 14–20 and 10–16 CFU/g, respectively. A total of 46 morphologically different bacterial strains were identified and assigned under the phylum Firmicutes. Identified bacterial strains belonged to the genus Bacillus and Staphylococcus and majority of the isolates were Bacillus subtilis and Staphylococcus nepalensis. Bacterial isolate HNS60 isolated from Hentak and identified as Bacillus subtilis was shown to possess high antimicrobial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Most of the identified bacteria were shown to possess DPPH radical scavenging as well as phosphatase, amylase, protease and cellulose activities. The isolate HNS60 contain high antimicrobial, enzymatic and probiotics activity which might responsible for the possible health benefits of the fermented foods. Utonga-kupsu, Hentak and Ngari thus can be further exploited as a potential source of probiotics and natural antioxidants.


Fermented fish 16s rRNA Antimicrobial Enzymatic activities Probiotics 



This research was supported by a grant from the Advanced Level State Biotech Hub sponsored by Department of Biotechnology (DBT), Govt. of India, New Delhi. The authors also thank the DBT-Bioinformatics Infrastructure and DeLCON e-journal facilities which were utilized for this study.

Supplementary material

13197_2018_3103_MOESM1_ESM.docx (25 kb)
Supplementary material 1 (DOCX 24 kb)
13197_2018_3103_MOESM2_ESM.doc (85 kb)
Supplementary material 2 (DOC 85 kb)


  1. Angmo K, Kumari A, Bhalla TC (2016) Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of Ladakh. LWT Food Sci Technol 66:428–435CrossRefGoogle Scholar
  2. Caplice E, Fitzgerald GF (1999) Food fermentations: role of microorganisms in food production and preservation. Int J Food Microbiol 50(1):131–149CrossRefGoogle Scholar
  3. Casaburi A, Blaiotta G, Mauriello G, Pepe O, Villani F (2005) Technological activities of Staphylococcus carnosus and Staphylococcus simulans strains isolated from fermented sausages. Meat Sci 71(4):643–650CrossRefGoogle Scholar
  4. Chettri R, Tamang JP (2015) Bacillus species isolated from Tungrymbai and Bekang, naturally fermented soybean foods of India. Int J Food Microbiol 197:72–76CrossRefGoogle Scholar
  5. Crisan EV, Sands A (1975) Microflora of four fermented fish sauces. Appl Microbiol 29(1):106–108Google Scholar
  6. DeLong EF, Pace NR (2001) Environmental diversity of bacteria and archaea. Syst Biol 50(4):470–478CrossRefGoogle Scholar
  7. Drosinos EH, Paramithiotis S, Kolovos G, Tsikouras I, Metaxopoulos I (2007) Phenotypic and technological diversity of lactic acid bacteria and staphylococci isolated from traditionally fermented sausages in Southern Greece. Food Microbiol 24(3):260–270CrossRefGoogle Scholar
  8. Einarsson H, Lauzon HL (1995) Biopreservation of brined shrimp (Pandalus borealis) by bacteriocins from lactic acid bacteria. Appl Environ Microbiol 61(2):669–676Google Scholar
  9. Holmes-Farley S, Burke S (2002) Inventors; Geltex Pharmaceutical, Inc., assignee. Method for treating gout and reducing serum uric acid. United States patent application US 10/125,793Google Scholar
  10. Holt JG, Krieg NR, Sneath PH, Staley JT, Williams ST (1994) Bergey’s manual of determinate bacteriology. ISBN 0-683-00603-7Google Scholar
  11. Jeyaram K, Singh TA, Romi W, Devi AR, Singh WM, Dayanidhi H, Singh NR, Tamang JP (2009) Traditional fermented foods of Manipur. Indian J Tradit Knowl 8(1):115–121Google Scholar
  12. Krauss BH (1993) Plants in Hawaiian culture. University of Hawaii Press, HonoluluGoogle Scholar
  13. Liasi SA, Azmi TI, Hassan MD, Shuhaimi M, Rosfarizan M, Ariff AB (2009) Antimicrobial activity and antibiotic sensitivity of three isolates of lactic acid bacteria from fermented fish product, Budu. Malays J Microbiol 5(1):33–37Google Scholar
  14. Macfarlane S, Macfarlane GT (2006) Composition and metabolic activities of bacterial biofilms colonizing food residues in the human gut. Appl Environ Microbiol 72(9):6204–6211CrossRefGoogle Scholar
  15. Maga EA, Desai PT, Weimer BC, Dao N, Kültz D, Murray JD (2012) Consumption of lysozyme-rich milk can alter microbial fecal populations. Appl Environ Microbiol 78(17):6153–6160CrossRefGoogle Scholar
  16. Majumdar RK, Basu S (2010) Characterization of the traditional fermented fish product Lonailish of Northeast India. Indian J Tradit Knowl 9:453–458Google Scholar
  17. Majumdar RK, Roy D, Bejjanki S, Bhaskar N (2016) Chemical and microbial properties of shidal, a traditional fermented fish of Northeast India. J Food Sci Technol 53(1):401–410CrossRefGoogle Scholar
  18. Miralles MC, Flores J, Perez-Martinez G (1996) Biochemical tests for the selection of Staphylococcusstrains as potential meat starter cultures. Food Microbiol 13(3):227–236CrossRefGoogle Scholar
  19. O’Toole GA, Pratt LA, Watnick PI, Newman DK, Weaver VB, Kolter R (1999) Genetic approaches to study of biofilms. Methods Enzymol 310:91–109CrossRefGoogle Scholar
  20. Parvez S, Malik KA, Ah Kang S, Kim HY (2006) Probiotics and their fermented food products are beneficial for health. J Appl Microbiol 100(6):1171–1185CrossRefGoogle Scholar
  21. Peralta EM, Hatate H, Kawabe D, Kuwahara R, Wakamatsu S, Yuki T, Murata H (2008) Improving antioxidant activity and nutritional components of Philippine salt-fermented shrimp paste through prolonged fermentation. Food Chem 111(1):72–77CrossRefGoogle Scholar
  22. Phadke G, Elavarasan K, Shamasundar BA (2014) Angiotensin-I converting enzyme (ACE) inhibitory activity and antioxidant activity of fermented fish product ngari as influenced by fermentation period. Int J Pharm Bio Sci 5(2):134–142Google Scholar
  23. Ramos HC, Hoffmann T, Marino M, Nedjari H, Presecan-Siedel E, Dreesen O, Glaser P, Jahn D (2000) Fermentative metabolism of Bacillus subtilis: physiology and regulation of gene expression. J Bacteriol 182(11):3072–3080CrossRefGoogle Scholar
  24. Salehi S, Eckley L, Sawyer GJ, Zhang X, Dong X, Freund JN, Fabre JW (2009) Intestinal lactase as an autologous β-galactosidase reporter gene for in vivo gene expression studies. Hum Gene Ther 20(1):21–30CrossRefGoogle Scholar
  25. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. Cold Spring Harbor Laboratory Press, New York. ISBN 0879693096Google Scholar
  26. Sarojnalini CH, Suchitra T (2009) Microbial profile of starter culture fermented fish product ‘Ngari’ of Manipur. Indian J Fish 56(2):123–127Google Scholar
  27. Singh KK, Shankar PD, Abdhul KA (2015) Study of microbial diversity for bacteriocin production from NGARI fish and noble synthesis of silver nanoparticles and its antimicrobial activities. World J Pharm Sci 5(3):953–974Google Scholar
  28. Tamang JP, Tamang N, Thapa S, Dewan S, Tamang B, Yonzan H, Rai AK, Chettri R, Chakrabarty J, Kharel N (2012) Microorganisms and nutritional value of ethnic fermented foods and alcoholic beverages of North East India. Indian J Tradit Knowl 11(1):7–25Google Scholar
  29. Tamang JP, Watanabe K, Holzapfel WH (2016) Review: diversity of microorganisms in global fermented foods and beverages. Front Microbiol. Google Scholar
  30. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28(10):2731–2739CrossRefGoogle Scholar
  31. Tanasupawat S, Ezaki T, Suzuki KI, Okada S, Komagata K, Kozaki M (1992) Characterization and identification of Lactobacillus pentosus and Lactobacillus plantarum strains from fermented foods in Thailand. J Gen Appl Microbiol 38(2):121–134CrossRefGoogle Scholar
  32. Thapa N (2016) Ethnic fermented and preserved fish products of India and Nepal. J Ethnic Foods 3(1):69–77CrossRefGoogle Scholar
  33. Thapa N, Pal J, Tamang JP (2004) Microbial diversity in ngari, hentak and tungtap, fermented fish products of North-East India. World J Microbiol Biotechnol 20(6):599–607CrossRefGoogle Scholar
  34. Thapa N, Pal J, Tamang JP (2006) Phenotypic identification and technological properties of lactic acid bacteria isolated from traditionally processed fish products of the Eastern Himalayas. Int J Food Microbiol 107(1):33–38CrossRefGoogle Scholar
  35. Vizoso-Pinto MG, Franz CMAP, Schillinger U, Holzapfel WH (2006) Lactobacillus spp. within vitro probiotic properties from human faeces and traditional fermented products. Int J Food Microbiol 109(3):205–214CrossRefGoogle Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2018

Authors and Affiliations

  1. 1.Department of BiotechnologyMizoram UniversityAizawlIndia

Personalised recommendations