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Effect of bovine lactoferrin on the internalization of coagulase-negative staphylococci into bovine mammary epithelial cells under in-vitro conditions

Published online by Cambridge University Press:  05 January 2009

Paula Hyvönen ,
Sari Käyhkö ,
Suvi Taponen ,
Atte von Wright  and
Satu Pyörälä
Paula Hyvönen*
Affiliation:
Department of Biosciences, University of Kuopio, PO Box 1627, FI-70211 Kuopio, Finland
Sari Käyhkö
Affiliation:
Department of Biosciences, University of Kuopio, PO Box 1627, FI-70211 Kuopio, Finland
Suvi Taponen
Affiliation:
Department of Clinical Veterinary Science, Saari Unit, Faculty of Veterinary Medicine, University of Helsinki, FI-04920 Saarentaus, Finland
Atte von Wright
Affiliation:
Department of Biosciences, University of Kuopio, PO Box 1627, FI-70211 Kuopio, Finland
Satu Pyörälä
Affiliation:
Department of Clinical Veterinary Science, Saari Unit, Faculty of Veterinary Medicine, University of Helsinki, FI-04920 Saarentaus, Finland
*
*For correspondence; e-mail: paula.hyvonen@uku.fi
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Abstract

Coagulase-negative staphylococci (CNS) have emerged as bovine mastitis pathogens in many countries. CNS mastitis is generally mild but can persist in the udder for long periods. Pathogenesis of CNS intramammary infection is not well understood. In the present study, adhesion, invasion and intracellular replication of twenty-two CNS strains isolated from bovine mastitis and the effect of bovine lactoferrin (bLf) on the internalization were studied in vitro in a bovine mammary epithelial (BME) cell model. The CNS strains were of Staphylococcus chromogenes, Staph. simulans, Staph. epidermidis, Staph. haemolyticus and Staph. cohnii urealyticus; two strains of Staph. aureus were used as controls. Seven of the CNS strains originated from persistent and five from transient mastitis infections. The in-vitro susceptibility of the strains to bLf was also investigated. All CNS species examined had an adhesive ability equal to that of Staph. aureus, but internalization varied among staphylococcal strains. The antagonistic effect of bLf on the adhesion and invasion of CNS strains was weak, but bLf significantly decreased intracellular replication and replication rates of CNS. No correlation between the in-vitro susceptibility of the strain to bLf or internalization among clinical signs of mastitis was established. No difference between the persistent and transient CNS strains in adhesion, invasion or replication rate was recorded. This in-vitro BME cell model can be used to study the virulence potential of mastitis pathogens, although the severity and persistence of eventual infections shall be further investigated in vivo. The role of bLf in intramammary infection caused by CNS may be limited.

Keywords

Coagulase-negative staphylococcibovine lactoferrininternalisationmammary epithelial cells
Type
Research Article
Information
Journal of Dairy Research , Volume 76 , Issue 2 , May 2009 , pp. 144 - 151
Copyright
Copyright © Proprietors of Journal of Dairy Research 2008

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References

Aarestrup, FM & Jensen, NE 1997 Prevalence and duration of intramammary infection in Danish heifers during peripartim period. Journal of Dairy Science 80 307–312Google Scholar
Aarestrup, FM, Larsen, HD & Jensen, NE 1999 Characterization of Staphylococcus simulans strains isolated from cases of bovine mastitis. Veterinary Microbiology 66 165–170CrossRefGoogle ScholarPubMed
Aguilar, B, Amorena, B & Iturralde, M 2001 Effect of slime on adherence of Staphylococcus aureus isolated from bovine and ovine mastitis. Veterinary Microbiology 78 183–191Google Scholar
Almeida, RA, Matthews, KR, Cifrian, E, Guidry, AJ & Oliver, SP 1996 Staphylococcus aureus invasion of bovine mammary epithelial cells. Journal of Dairy Science 79 1021–1026CrossRefGoogle ScholarPubMed
Almeida, RA & Oliver, SP 2001 Interaction of coagulase-negative staphylococcus species with bovine mammary epithelial cells. Microbial Pathogenesis 31 205–212Google ScholarPubMed
Alugupalli, KR & Kalfas, S 1997 Characterization of the lactoferrin-dependent inhibition of the adhesion of Actinobacillus actinomycetemcomitans, Prevotella intermedia and Prevotella nigrecens to fibroblasts and to a reconstituted basement membrane. Acta Pathologica, Microbiologica et Immunologica 105 680–688Google Scholar
Anaya-Lopez, JL, Contreras-Guzman, OE, Carabez-Trejo, A, Baizabal-Aguirre, VM, Lopez-Meza, JE, Valdez-Alarcon, JJ & Ochoa-Zarzosa, A 2006 Invasive potential of bacterial isolates associated with subclinical bovine mastitis. Research in Veterinary Science 81 358–361Google Scholar
Berlutti, F, Schippa, S, Morea, C, Sarli, S, Perfetto, B, Donnarumma, G & Valenti, P 2006 Lactoferrin downregulates pro-inflammatory upexpressed in intestinal epithelial cells infected with invasive or noninvasive Escherichia coli strains. Biochemistry and Cell Biology 84 351–357Google Scholar
Bradley, AJ, Leach, KA, Breen, JE, Green, LE & Green, MJ 2007 Survey of the incidence and aetiology of mastitis on dairy farms in England and Wales. Veterinary Record 160 253–257Google Scholar
Brock, JH 2002 The physiology of lactoferrin. Biochemistry and Cell Biology 80 1–6Google Scholar
Chaffer, M, Leitner, G, Winkler, M, Glickman, A, Krifucks, O, Ezra, E & Saran, A 1999 Coagulase-negative staphylococci and mammary gland infections in cows. Zentralblatt für Veterinärmedizin B 46 707–712Google Scholar
Cifrian, E, Guidry, AJ, O'Brien, CN, Nickerson, SC & Marquardt, WW 1994 Adherence of Staphylococcus aureus to cultured bovine mammary epithelial cells. Journal of Dairy Science 77 970–983CrossRefGoogle ScholarPubMed
Cucarella, C, Tormo, MA, Ubeda, C, Trotonda, MP, Monzon, M, Peris, C, Amorena, B, Lasa, I & Penades, JR 2004 Role of biofilm-associated protein bap in the pathogenesis of bovine Staphylococcus aureus. Infection and Immunity 72 2177–2185Google Scholar
de Lillo, A, Quiros, LM & Fierro, JF 1997 Relationship between antibacterial activity and cell surface binding of lactoferrin in species of genus Micrococcus. FEMS Microbiology Letters 150 89–94CrossRefGoogle ScholarPubMed
Diarra, MS, Petitclerc, D, Deschenes, E, Lessard, N, Grondin, G, Talbot, BG & Lacasse, P 2003 Lactoferrin against Staphylococcus aureus mastitis. Lactoferrin alone or in combination with penicillin G on bovine polymorphonuclear function and mammary epithelial cells colonisation by Staphylococcus aureus. Veterinary Immunology and Immunopathology 95 33–42CrossRefGoogle ScholarPubMed
Dogan, B, Klaessig, S, Rishniw, M, Almeida, RA, Oliver, SP, Simpson, K & Schukken, YH 2006 Adherent and invasive Escherichia coli are associated with persistent bovine mastitis. Veterinary Microbiology 116 270–282Google Scholar
Finlay, BB & Cossart, P 1997 Exploitation of mammalian host cell functions by bacterial pathogens. Science 276 718–725Google Scholar
Griesbeck-Zilch, B, Meyer, HHD, Kühn, Ch, Schwerin, M & Wellnitz, O 2008 Staphylococcus aureus and Escherichia coli cause deviating expression profiles of cytokines and lactoferrin messenger ribonucleic acid in mammary epithelia cells. Journal of Dairy Science 91 2215–2224Google Scholar
Haltia, L, Honkanen-Buzalski, T, Spiridonova, I, Olkonen, A & Myllys, V 2006 A study of bovine mastitis, milking procedures and management practices on 25 Estonian dairy herds. Acta Veterinary Scandinavica 48 22–28CrossRefGoogle ScholarPubMed
Hebert, A, Sayasith, K, Senechal, S, Dubreuil, P & Lagace, J 2000 Demonstration of intracellular Staphylococcus aureus in bovine mastitis alveolar cells and macrophages isolated from naturally infected cow milk. FEMS Microbiology Letters 193 57–62CrossRefGoogle ScholarPubMed
Hensen, SM, Pavicic, MJ, Lohuis, JA & Poutrel, B 2000 Use of bovine primary mammary epithelial cells for the comparison of adherence and invasion ability of Staphylococcus aureus strains. Journal of Dairy Science 83 418–429CrossRefGoogle ScholarPubMed
Hyvonen, P, Suojala, L, Orro, T, Haaranen, J, Simola, O, Rontved, C & Pyorala, S 2006 Transgenic cows that produce recombinant human lactoferrin in milk are not protected from experimental Escherichia coli intramammary infection. Infection and Immunity 74 6206–6212CrossRefGoogle Scholar
Jarp, J 1991 Classification of coagulase-negative staphylococci isolated from bovine clinical and subclinical mastitis. Veterinary Microbiology 27 151–158Google Scholar
Karpulus, TE, Ulevitch, RJ & Wilson, CB 1987 A new method for reduction of endotoxin contamination from protein solutions. Journal of Immunological Methods 105 211–220CrossRefGoogle Scholar
Kawai, K, Hagiwara, S, Anri, A & Nagahata, H 1999 Lactoferrin concentration in milk of bovine clinical mastitis. Veterinary Research Communications 23 391–398Google Scholar
Kerro Dego, O, van Dijk, JE & Nederbragt, H 2002 Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review. Veterinary Quarterly 24 181–198CrossRefGoogle ScholarPubMed
Kloos, WE & Bannerman, TL 1994 Update on clinical significance of coagulase-negative staphylococci. Clinical Microbiology Review 7 117–140Google Scholar
Komine, K, Komine, Y, Kuroishi, T, Kobayashi, J, Obara, Y & Kumagai, K 2005 Small molecule lactoferrin with an inflammatory effect but no apparent antibacterial activity in mastitic mammary gland secretion. Journal of Veterinary Medical Science 67 667–677CrossRefGoogle ScholarPubMed
Lee, NY, Kawai, K, Nakamura, I, Tanaka, T, Kumura, H & Shimazaki, K 2004 Susceptibilities against bovine lactoferrin with microorganisms isolated from mastitic milk. Journal of Veterinary Medical Science 66 1267–1269Google ScholarPubMed
Lonnerdal, B & Iyer, S 1995 Lactoferrin: Molecular structure and biological function. Annual Review of Nutrition 15 93–110CrossRefGoogle ScholarPubMed
Mamo, W, Froman, G & Wadstrom, T 1988 Interaction of sub-epithelial connective tissue components with Staphylococcus aureus and coagulase-negative staphylococci from bovine mastitis. Veterinary Microbiology 18 163–176Google Scholar
Matthews, KR, Almeida, RA & Oliver, SP 1994 Bovine mammary epithelial cell invasion by Streptococcus uberis. Infection and Immunity 62 5641–5646CrossRefGoogle ScholarPubMed
Melchior, MB, Vaarkamp, H & Fink-Grimmels, J 2006 Biofilms: S role in recurrent mastitis infections? The Veterinary Journal 171 398–407Google Scholar
Naidu, AS, Andersson, M, Miedzobrodzki, J, Forsgren, A & Watts, JL 1991 Bovine lactoferrin receptors in Staphylococcus aureus isolated from bovine mastitis. Journal of Dairy Science 74 1218–1226Google Scholar
Naidu, AS, Miedzobrodzki, J, Andersson, M, Nilsson, LE, Forsgren, A & Watts, JL 1990 Bovine lactoferrin binding to six species of coagulase-negative staphylococci isolated from bovine intramammary infections. Journal of Clinical Microbiology 28 2312–2319Google Scholar
Nevala, M, Taponen, S & Pyörälä, S 2004 Bacterial etiology of bovine clinical mastitis – data from Saari ambulatory clinic in 2002–2003. Suomen Eläinlääkärilehti 110 363–369Google Scholar
Oliveira, M, Bexiga, R, Nunes, SF, Carneiro, C, Cavaco, LM, Bernardo, F & Vilela, CL 2006 Biofilm-forming ability profiling of Staphylococcus aureus and Staphylococcus epidermidis mastitis isolates. Veterinary Microbiology 118 133–140CrossRefGoogle ScholarPubMed
Pitkala, A, Haveri, M, Pyorala, S, Myllys, V & Honkanen-Buzalski, T 2004 Bovine mastitis in Finland 2001 – prevalence, distribution of bacteria, and antimicrobial resistance. Journal of Dairy Science 87 2433–2441Google Scholar
Rainard, P 1986 Bacteriostatic activity of bovine milk lactoferrin against mastitic bacteria. Veterinary Microbiology 11 387–392Google Scholar
Rainard, P 1992 Binding of bovine lactoferrin to Streptococcus agalactiae. FEMS Microbiology Letters 77 235–239Google Scholar
Rainard, P & Riollet, C 2006 Innate immunity of the bovine mammary gland. Veterinary Research 37 369–400Google Scholar
Rejman, JJ, Turner, JD & Oliver, SP 1994 Characterization of lactoferrin binding to the MAC-T bovine mammary epithelial cell line using a biotin-avidin technique. International Journal of Biochemistry 26 201–206CrossRefGoogle Scholar
Rochard, E, Legrand, D, Lecocq, M, Hamelin, R, Crepin, M, Montreuil, J & Spik, G 1992 Characterization of lactotransferrin receptor in epithelial cell lines from non-malignant human breast, benign mastopathies and breast carcinomas. Anticancer Research 12 2047–2051Google Scholar
Singh, PK, Parsek, MR, Greenberg, EP & Welsh, MJ 2002 A component of innate immunity prevents bacterial biofilm development. Nature 417 552–555Google Scholar
Taponen, S, Koort, J, Bjorkroth, J, Saloniemi, H & Pyorala, S 2007 Bovine intramammary infections caused by coagulase-negative staphylococci may persist throughout lactation according to amplified fragment length polymorphism-based analysis. Journal of Dairy Science 90 3301–3307Google Scholar
Taponen, S, Simojoki, H, Haveri, M, Larsen, HD & Pyorala, S 2006 Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP. Veterinary Microbiology 115 199–207CrossRefGoogle ScholarPubMed
Tenhagen, BA, Koster, G, Wallmann, J & Heuwieser, W 2006 Prevalence of mastitis pathogens and their resistance against antimicrobial agents in dairy cows in Brandenburg, Germany. Journal of Dairy Science 89 2542–2551CrossRefGoogle ScholarPubMed
Van Belkum, A, Kools-Sijmons, M & Verbrugh, H 2002 Attachment of Staphylococcus aureus to eukaryotic cells and experimental pitfalls in staphylococcal adherence assays: A critical appraisal. Journal of Microbiological Methods 48 19–42Google Scholar
Van Oostveldt, K, Tomita, GM, Paape, MJ, Capuco, AV & Burvenich, C 2002 Apoptosis of bovine neutrophils during mastitis experimentally induced with Escherichia coli or endotoxin. American Journal of Veterinary Research 63 448–453Google ScholarPubMed
Vasudevan, P, Nair, MKN, Thirunavukkarasu, A & Venkitanarayanan, KS 2003 Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Vetrinary Microbiology 92 179–185Google Scholar
Waage, S, Mork, T, Roros, A, Aasland, D, Hunshamar, A & Odegaard, SA 1999 Bacteria associated with clinical mastitis in dairy heifers. Journal of Dairy Science 82 712–719Google Scholar
Zhang, S & Maddox, CW 2000 Cytotoxic activity of coagulase-negative staphylococci in bovine mastitis. Infection and Immunity 68 1102–1108Google Scholar