Isolation and characterization of a Bacillus spp. for manufacturing the feed additives in livestock

가축의 보조사료 개발을 위한 Bacillus spp.의 분리 및 특성

  • Park, Hae Suk (Microbial Institute for Fermentation Industry) ;
  • Jo, Seung Wha (Microbial Institute for Fermentation Industry) ;
  • Yim, Eun Jung (Microbial Institute for Fermentation Industry) ;
  • Kim, Yun Sun (Microbial Institute for Fermentation Industry) ;
  • Moon, Sung Hyun (College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University) ;
  • Cho, Ho Seong (College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University) ;
  • Kim, Hyun-Young (Institute of Sunchang Fermented Soybean Products) ;
  • Cho, Yong Sik (Fermented Food Science Division, National Academy of Agricultural Science, RDA) ;
  • Cho, Sung Ho (Microbial Institute for Fermentation Industry)
  • 박해석 (재단법인발효미생물산업진흥원) ;
  • 조승화 (재단법인발효미생물산업진흥원) ;
  • 임은정 (재단법인발효미생물산업진흥원) ;
  • 김윤순 (재단법인발효미생물산업진흥원) ;
  • 문성현 (전북대학교 수의학과 및 생체안전성연구소) ;
  • 조호성 (전북대학교 수의학과 및 생체안전성연구소) ;
  • 김현영 (순창장류사업소) ;
  • 조용식 (농촌진흥청 국립농업과학원 발효식품과) ;
  • 조성호 (재단법인발효미생물산업진흥원)
  • Received : 2015.11.27
  • Accepted : 2015.12.21
  • Published : 2015.12.31


The aims of this study were to isolate spore-forming Bacillus strains that exhibit high digestibility and anti-pathogenic bacteria toward feed for calves. Total 136 spore-forming strains were isolated from finished feeds and their ingredients. Among them, 93 strains were identified as Bacillus species when analyzed by 16S rRNA sequencing. For industrial use, three strains named as Bacillus licheniformis SHS14, B. subtilis LCB7, B. amyloliquefaciens LCB10 were selected after evaluating the industrial standards that are related with heat and acid resistance, enzyme activities, and anti-pathogenic activities against Samonella dublin ATCC15480 and E. coli K99. After each culture, 3 selected strains were mixed together at 1:1:1 (v/v/v) ratio and then prepared as the mixed starter culture for feeding. The changes in microbial community were analyzed via 16S rRNA metagenomics. The initial community ratio among three strains was maintained even after manufacturing into final products. Also, in vitro, enzymatic and anti-pathogenic activities were almost same as those when cultured in single culture, and results of anti-pathogenic activities conducted with calves showed 90% activities against lincomycin, which would be indicative of a promising feed starter.


Bacillus;fermented feed;spore forming bacteria;starter culture


Supported by : 농촌진흥청


  1. Banihashemi, A., Dyke, M., and Huck, P. 2012. Long-amplicon propidium monoazide - PCR enumeration assay to detect viable Campylobacter and Salmonella. J. Appl. Microbiol. 113, 863-873.
  2. De Clerck, E., Gevers, D., De Ridder, K., and De Vos, P. 2004. Screening of bacterial contamination during gelatin production by means of denaturing gradient gel electrophoresis, focused on Bacillus and related endospore-forming genera. J. Appl. Microbiol. 96, 1333-1341.
  3. EFSA Panel on Biological Hazards (BIOHAZ). 2010. Scientific opinion on the maintenance of the list of QPS biological agents intentionally added to food and feed. EFSA J. 8, 1944.
  4. Kang, M.I., Han, D.U., Chung, Y.U., Chung, D.Y., Lee, C.Y., Lee, C.G., Wee, S.H., and Cho, J.J. 2001. Survey on Korean-native calves diseases and mortality. Korean J. Vet. Serv. 24, 223-241.
  5. Kim, J.W., Jun, K.D., Kang, J.S., Jang, J.S., Ha, B.J., and Lee, J.H. 2005. Characterization of Bacillus licheniformis as a probiotic. Korean J. Biotechnol. Bioeng. 20, 359-362.
  6. Kim, K.H. and Song, M.K. 2013. Effects of feeding level of feedstuffs fermented with complex probiotics on growth of Holstein male calves. Korean Bull. Animal Biotechnol. 5, 1-9.
  7. Larsen, N., Thorsen, L., Kpikpi, E.N., Lauridsen, B.S., Cantor, M.D., Nielsen, B., Brockmann, E., Derkx, P.M.F., and Jespersen, L. 2014. Characterization of Bacillus spp. strains for use as probiotic additives in pig feed. Appl. Microbiol. Biotechnol. 98, 1105-1118.
  8. Leser, T.D., Knarreborg, A., and Worm, J. 2008. Germination and outgrowth of Bacillus subtilis and Bacillus licheniformis spores in the gastrointestinal tract of pigs. J. Appl. Microbiol. 104, 1025-1033.
  9. Paik, H.D., Jung, M.Y., Jung, H.Y., Kim, W.S., and Kim, K.T. 2002. Characterization of Bacillus polyfermenticus SCD for oral bacteriotherapy of gastrointestinal disorders. Korean J. Food. Sci. Technol. 34, 73-78.
  10. Sanders, M.E., Morelli, L., and Tompkins, A. 2003. Sporeformers as human probiotics: Bacillus, Sporolactobacillus, and Brevibacillus. Comp. Rev. Food Sci. Food Safety 2, 101-110.
  11. Shin, T.S. 2014. Development of supplementary fodder as an alternative of antibiotics using by heat- resistance microbes. Busan National University.
  12. Szczawinska, M.E., Thayer, D.W., and Phillips, J.G. 1991. Fate of unirradiated Salmonella in irradiated mechanically deboned chicken meat. Int. J. Food Microbiol. 14, 313-324.
  13. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S. 2011. MEGA5 : Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Cell 28, 2731-2739.
  14. Veith, B., Herzberg, C., Steckel, S., Feesche, J., Maurer, K.H., Ehrenreich, P., Baumer, S., Henne, A., Liesegang, H., Merkl, R., et al. 2004. The complete genome sequence of Bacillus licheniformis DSM13, an organism with great industrial potential. J. Mol. Microbiol. Biotechnol. 7, 204-211.
  15. Yousif, A.A., Mahmood, N.M., and Al-Taai, N.A. 2013. Immunization of mice with killed E. coli K99 vaccine for protection against colibaccillosis. Int. J. Microbiol. Res. 5, 482-485.