Abstract
Various bacteria have been found in raw cow’s milk, and identifying milk microflora and its functions is critical for maintaining cow health and farm hygiene. Although studies on pathogens and spoilage bacteria in milk have been widely reported, the relationship between milk bacteria, including nonpathogenic bacteria, and the bovine udder is poorly understood. We investigated milk microflora over 1 year using a culture-dependent method and culture-independent analysis by denaturing gradient gel electrophoresis. Among 240 isolates, Lactococcus lactis (81/240) was predominant. The predominant genera were Lactococcus, Stenotrophomonas, Microbacterium, Chryseobacterium, Serratia and Pseudomonas. Among seven strains belonging to these predominant genera, two strains of L. lactis (ssp. lactis and ssp. cremoris) exhibited the highest adherence to bovine mammary gland epithelial cells (BMECs) derived from the bovine udder; 3.4 % of the inoculated bacteria adhered to BMECs. This was followed by Serratia sp. (1.6 %), Microbacterium sp. (0.8 %), Stenotrophomonas maltophilia (0.5 %), Pseudomonas sp. (0.3 %) and Chryseobacterium sp. (0.1 %). The two L. lactis isolates exhibited higher adherence to BMECs than type strains and isolates of various origins.
Similar content being viewed by others
References
Almeida RA, Luther DA, Park HM, Oliver SP (2006) Identification, isolation, and partial characterization of a novel Streptococcus uberis adhesion molecule (SUAM). Vet Microbiol 115:183–191. doi:10.1016/j.vetmic.2006.02.005
Barkema HW, Green MJ, Bradley AJ, Zadoks RN (2009) Invited review: the role of contagious disease in udder health. J Dairy Sci 92:4717–4729. doi:10.3168/jds.2009-2347
Beecher C, Daly M, Berry DP, Klostermann K, Flynn J, Meaney W, Hill C, McCarthy TV, Ross RP, Giblin L (2009) Administration of a live culture of Lactococcus lactis DPC 3147 into the bovine mammary gland stimulates the local host immune response, particularly IL-1β eand IL-8 gene expression. J Dairy Res 76:340–348. doi:10.1017/S0022029909004154
Cogan TM, Beresford TP, Steele J, Broadbent J, Shah NP, Ustunol Z (2007) Invited review: advances in starter cultures and cultured foods. J Dairy Sci 90:4005–4021. doi:10.3168/jds.2006-765
Delbes C, Ali-Mandjee L, Montel MC (2007) Monitoring bacterial communities in raw milk and cheese by culture-dependent and -independent 16S rRNA gene-based analyses. Appl Environ Microbiol 73:1882–1891. doi:10.1128/AEM.01716-06
Dieckelmann M, Johnson LA, Beacham IR (1998) The diversity of lipases from psychrotrophic strains of Pseudomonas: a novel lipase from a highly lipolytic strain of Pseudomonas fluorescens. J Appl Microbiol 85:527–536. doi:10.1046/j.1365-2672.1998.853530.x
Dillon RJ, Dillon VM (2004) The gut bacteria of insects: nonpathogenic interactions. Annu Rev Entomol 49:71–92. doi:10.1146/annurev.ento.49.061802.123416
Edelman S, Westerlund-Wikström B, Leskelä S, Kettunen H, Rautonen N, Apajalahti J, Korhonen TK (2002) In vitro adhesion specificity of indigenous Lactobacilli within the avian intestinal tract. Appl Environ Microbiol 68:5155–9. doi:10.1128/AEM.68.10.5155-5159.2002
Espeche MC, Otero MC, Sesma F, Nader-Macias ME (2009) Screening of surface properties and antagonistic substances production by lactic acid bacteria isolated from the mammary gland of healthy and mastitic cows. Vet Microbiol 135:346–357. doi:10.1016/j.vetmic.2008.09.078
Hagi T, Kobayashi M, Nomura M (2010) Molecular-based analysis of changes in indigenous milk microflora during the grazing period. Biosci Biotech Biochem 74:484–487. doi:10.1271/bbb.90470
Hagi T, Tanaka D, Iwamura Y, Hoshino T (2004) Diversity and seasonal changes in lactic acid bacteria in the intestinal tract of cultured freshwater fish. Aquaculture 234:335–346. doi:10.1016/j.aquaculture.2004.01.018
Hantsis-Zacharov E, Halpern M (2007) Culturable psychrotrophic bacterial communities in raw milk and their proteolytic and lipolytic traits. Appl Environ Microbiol 73:7162–7168. doi:10.1128/AEM.00866-07
Human Microbiome Jumpstart Reference Strains Consortium (2010) A catalog of reference genomes from the human microbiome. Science 328:994–999. doi:10.1126/science.1183605
Ibeagha-Awemu EM, Lee JW, Ibeagha AE, Bannerman DD, Paape MJ, Zhao X (2008) Bacterial lipopolysaccharide induces increased expression of toll-like receptor (TLR) 4 and downstream TLR signaling molecules in bovine mammary epithelial cells. Vet Res 39:11. doi:10.1051/vetres:2007047
Ismail AS, Hooper LV (2005) Epithelial cells and their neighbors. IV. Bacterial contributions to intestinal epithelial barrier integrity. Am J Physiol Gastrointest Liver Physiol 289:G779–784. doi:10.1152/ajpgi.00203.2005
Kimoto H, Kurisaki J, Tsuji NM, Ohmomo S, Okamoto T (1999) Lactococci as probiotic strains: adhesion to human enterocyte-like Caco-2 cells and tolerance to low pH and bile. Lett Appl Microbiol 29:313–316. doi:10.1046/j.1365-2672.1999.00627.x
Kline KA, Fälker S, Dahlberg S, Normark S, Henriques-Normark B (2009) Bacterial adhesins in host-microbe interactions. Cell Host Microbe 5:580–592. doi:10.1016/j.chom.2009.05.011
Klostermann K, Crispie F, Flynn J, Ross RP, Hill C, Meaney W (2008) Intramammary infusion of a live culture of Lactococcus lactis for treatment of bovine mastitis: comparison with antibiotic treatment in field trials. J Dairy Res 75:365–373. doi:10.1017/S0022029908003373
Kuang Y, Tani K, Synnott AJ, Ohshima K, Higuchi H, Nagahata H, Tanji Y (2009) Characterization of bacterial population of raw milk from bovine mastitis by culture-independent PCR–DGGE method. Biochem Eng J 45:76–81
Lafarge V, Ogier JC, Girard V, Maladen V, Leveau JY, Gruss A, Delacroix-Buchet A (2004) Raw cow milk bacterial population shifts attributable to refrigeration. Appl Environ Microbiol 70:5644–5650. doi:10.1128/AEM.70.9.5644-5650.2004
Moussavi M, Adams MC (2010) An in vitro study on bacterial growth interactions and intestinal epithelial cell adhesion characteristics of probiotic combinations. Curr Microbiol 60:327–335. doi:10.1007/s00284-009-9545-1
Muyzer G, de Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59:695–700
Nomura M, Kobayashi M, Narita T, Kimoto-Nira H, Okamoto T (2006) Phenotypic and molecular characterization of Lactococcus lactis from milk and plants. J Appl Microbiol 101:396–405. doi:10.1111/j.1365-2672.2006.02949.x
Oliver SP, Jayarao BM, Almeida RA (2007) Foodborne pathogens in milk and the dairy farm environment: Food safety and public health implications. Foodborne pathog Dis 2:115–129. doi:10.1089/fpd.2005.2.115
Pyörälä S (2003) Indicators of inflammation in the diagnosis of mastitis. Vet Res 34:565–578. doi:10.1051/vetres:2003026
Rainard P, Riollet C (2006) Innate immunity of the bovine mammary gland. Vet Res 37:369–400. doi:10.1051/vetres:2006007
Rose MT, Aso H, Yonekura S, Komatsu T, Hagino A, Ozutsumi K, Obara Y (2002) In vitro differentiation of a cloned bovine mammary epithelial cell. J Dairy Res 69:345–355. doi:10.1017/S0022029902005551
Scheldeman P, Herman L, Foster S, Heyndrickx M (2006) Bacillus sporothermodurans and other highly heat-resistant spore formers in milk. J Appl Microbiol 101:542–55. doi:10.1111/j.1365-2672.2006.02964.x
Slama P, Sladek Z, Rysanek D, Langrova T (2009) Effect of Staphylococcus aureus and Streptococcus uberis on apoptosis of bovine mammary gland lymphocytes. Res Vet Sci 87:233–238. doi:10.1016/j.rvsc.2009.03.005
Slama P, Sladek Z, Slamova R, Pavlik A (2011) Production of bovine IL-10 following stimulation with lipopolysaccharide and muramyl dipeptide. FEBS J 278:304
Takamatsu D, Hata E, Osaki M, Aso H, Kobayashi S, Sekizaki T (2008) Role of SraP in adherence of Staphylococcus aureus to the bovine mammary epithelia. J Vet Med Sci 70:735–738. doi:10.1292/jvms.70.735
Van Oostveldt K, Paape MJ, Burvenich C (2002) Apoptosis of bovine neutrophils following diapedesis through a monolayer of endothelial and mammary epithelial cells. J Dairy Sci 85:139–147. doi:10.3168/jds.S0022-0302(02)74062-9
Acknowledgments
The authors thank the staff of the Management Section, NARO Institute of Livestock and Grassland Science, Japan, for their handling of the animals. This work was supported in part by KAKENHI no. 23780273 Grant-in-Aid for Young Scientists (B).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hagi, T., Sasaki, K., Aso, H. et al. Adhesive properties of predominant bacteria in raw cow’s milk to bovine mammary gland epithelial cells. Folia Microbiol 58, 515–522 (2013). https://doi.org/10.1007/s12223-013-0240-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12223-013-0240-z