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Isolation and molecular characterization of lactobacilli from traditional fermented Dahi produced at different altitudes in Nepal

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Dairy Science & Technology

Abstract

Dairy products are an important part of daily food in the Himalayan country of Nepal. A wide variety of traditional fermented milk products are obtained in relation to different geo-climatic conditions of the country and different ethnic groups. Locally called Dahi is the most popular product, but little data are available on the autochthonous lactic acid bacteria (LAB) characterizing this yogurt-like product. Thirty-two replicate samples of indigenous Dahi were collected from four districts of Nepal at different altitude. In total of 193 isolates were obtained. Of these, the 120 rod-shaped isolates representing the dominant LAB population (62%) were further identified at the species level by using internal transcribed spacers (ITS) amplification, 16S rRNA gene sequence determination, and species-specific PCR. A further characterization at the strain level was carried out by combining analysis of repetitive elements and randomly amplified polymorphic DNA (RAPD) typing. Based on these analyses, the isolates were grouped in ten different species, among which Lactobacillus delbrueckii subsp. bulgaricus, L. paracasei, and L. rhamnosus represented the dominant species. A high degree of intraspecies diversity was also observed for all Lactobacillus species, except for L. rhamnosus isolates, which proved to give a single typing profile. Bacterial isolates represent a source of novel potential probiotics and starter cultures. The strain collection obtained from this study is a first step in the preservation of the natural biodiversity of bacterial population of the traditional Nepalese fermented Dahi.

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References

  • Bao Q, Yu J, Liu W, Qing M, Wang W, Chen X, Wang F, Li M, Wang H, Lv Q, Zhang H (2012) Predominant lactic acid bacteria in traditional fermented yak milk products in the Sichuan province of China. Dairy Sci Technol 92:309–319

    Article  CAS  Google Scholar 

  • Bhardwaj A, Puniya M, Sangu KPS, Kumar S, Dhewa T (2012) Isolation and biochemical characterization of Lactobacillus species isolated from Dahi. RRJoDST 1(2):18–31

    Google Scholar 

  • Colombo F, Borgo F, Fortina MG (2009) Genotypic characterization of non starter lactic acid bacteria involved in the ripening of artisanal Bitto PDO cheese. J Basic Microbiol 49:521–530

    Article  CAS  Google Scholar 

  • Coton M, Berthier F, Coton E (2008) Rapid identification of the three major species of dairy obligate heterofermenters Lactobacillus brevis, Lactobacillus fermentum and Lactobacillus parabuchneri by species-specific duplex PCR. FEMS Microbiol Lett 284:150–157

    Article  CAS  Google Scholar 

  • Dellaglio F, Felis GE, Castioni A, Torriani S, Germond JE (2005) Lactobacillus delbrueckii subsp. indicus subsp. nov., isolated from Indian dairy products. Int J Syst Evol Microbiol 55:401–404

    Article  CAS  Google Scholar 

  • Dewan S, Tamang JP (2007) Dominant lactic acid bacteria and their technological properties isolated from the Hymalayan ethnic fermented milk products. Ant van Leewen 92:343–352

    Article  CAS  Google Scholar 

  • Edwards U, Rogall T, Blöcker H, Emde M, Böttger EC (1989) Isolation and direct sequencing of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 17:7843–7853

    Article  CAS  Google Scholar 

  • Fortina MG, Ricci G, Mora D, Parini C, Manachini PL (2001) Specific identification of Lactobacillus helveticus by PCR with pepC, pepN and htrA targeted primers. FEMS Microbiol Lett 198:85–89

    Article  CAS  Google Scholar 

  • Guglielmetti S, Ciranna A, Mora D, Parini C, Karp M (2008) Construction, characterization and exemplificative application of bioluminescent Bifidobacterium longum biovar longum. Int J Food Microbiol 124(3):285–290

    Article  CAS  Google Scholar 

  • Heller KJ, Bockelmann W, Schrezenmeir J, de Vrese M (2003) Cheese and its potential as a probiotic food. In: Farnworth ER (ed) Handbook of fermented functional foods. CRC Press, Boca Raton, FL, pp 203–225

    Chapter  Google Scholar 

  • Jensen MA, Webster JA, Strauss N (1993) Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol 59:945–952

    CAS  Google Scholar 

  • Leroy F, DeVuyst L (2004) Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends Food Sci Technol 15:67–78

    Article  CAS  Google Scholar 

  • Mora D, Parini C, Fortina MG, Manachini PL (2000) Development of molecular RAPD marker for the identification of Pediococcus acidilactici strains. Syst Appl Microbiol 23:400–408

    Article  CAS  Google Scholar 

  • Rossetti L, Giraffa G (2005) Rapid identification of dairy lactic acid bacteria by M13-generated, RAPD-PCR fingerprint databases. J Microbiol Methods 63:135–144

    Article  CAS  Google Scholar 

  • Satish Kumar R, Kanmani P, Yuvaraj N, Paari KA, Pattukumar V, Arul V (2013) Traditional Indian fermented foods: a rich source of lactic acid bacteria. Int J Food Sci Nutr 64:415–428

    Article  CAS  Google Scholar 

  • Soomro AH, Masud T (2007) Protein pattern and plasmid profile of lactic acid bacteria isolated from dahi, a traditional fermented milk product of Pakistan. Food Technol Biotechnol 45(4):447–453

    CAS  Google Scholar 

  • Tamang JP, Dewan S, Thapa S, Olasupo NA, Schillinger U, Holzapfel WH (2000) Identification and enzymatic profiles of predominant lactic acid bacteria isolated from soft-variety chhurpi, a traditional cheese typical of the Sikkim Hiymalayas. Food Biotechnol 14:99–112

    Article  CAS  Google Scholar 

  • Tamang JP (2010) Himalayan fermented foods: microbiology, nutrition, and ethnic values. CRC Press, Taylor & Francis group

  • Taverniti V, Guglielmetti S (2010) Health-promoting properties of Lactobacillus helveticus. Front Microbiol 3:392. doi:10.3389/fmicb.2012.00392

    Google Scholar 

  • Torriani S, Felis GE, Dellaglio F (2001) Differentiation of Lactobacillus plantarum, L. pentosus, and L. paraplantarum by recA gene-derived primers. Appl Environ Microbiol 67:3450–3454

    Article  CAS  Google Scholar 

  • Torriani S, Zapparoli G, Dellaglio F (1999) Use of PCR-based methods for rapid differentiation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis. Appl Environ Microbiol 65(10):4351–4356

    CAS  Google Scholar 

  • Ward LJH, Timmins MJ (1999) Differentiation of Lactobacillus casei, L. paracasei and L. rhamnosus by polymerase chain reaction. Lett Appl Microbiol 29:90–92

    Article  CAS  Google Scholar 

  • Watanabe T, Hamada K, Tategaki A, Kishida H, Tanaka H, Kitano M, Miyamoto T (2009) Oral administration of lactic acid bacteria isolated from traditional South Asian fermented milk ‘dahi’ inhibits the development of atopic dermatitis in NC/Nga mice. J Nutr Sci Vitaminol (Tokyo) 55(3):271–278

    Article  CAS  Google Scholar 

  • Yu J, Wang WH, Menghe BLG, Jiri MT, Wang HM, Liu WJ, Bao QH, Lv Q, Zhang JC, Wang F, Xu HY, Sun TS, Zhang HP (2011) Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia. J. Dairy Sci 94:3229–3241

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study was financially supported by the Fondazione Cariplo (grant 2010–0678).

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Authors and Affiliations

  1. Department of Food, Environmental and Nutritional Sciences (DeFENS), Division of Food Microbiology and Bioprocessing, Università degli Studi di Milano, Via Celoria 2, 20133, Milan, Italy

    Giovanni Ricci, Valentina Taverniti, Chiara Ferrario, Maria Grazia Fortina & Simone Guglielmetti

  2. Nepal Academy of Science and Technology (NAST), Khumaltar, Lalitpur, Nepal

    Ranjan Koirala & Sangita Shrestha

  3. Central Department of Biotechnology Tribhuvan University, Kirtipur, Kathmandu, Nepal

    Ranjan Koirala & Rajani Malla

Authors
  1. Ranjan Koirala
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  2. Giovanni Ricci
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  3. Valentina Taverniti
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  4. Chiara Ferrario
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  5. Rajani Malla
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  6. Sangita Shrestha
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  7. Maria Grazia Fortina
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  8. Simone Guglielmetti
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Correspondence to Simone Guglielmetti.

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Koirala, R., Ricci, G., Taverniti, V. et al. Isolation and molecular characterization of lactobacilli from traditional fermented Dahi produced at different altitudes in Nepal. Dairy Sci. & Technol. 94, 397–408 (2014). https://doi.org/10.1007/s13594-014-0167-4

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  • DOI: https://doi.org/10.1007/s13594-014-0167-4

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