Abstract
Staphylococcus aureus is a common cause of intramammary infections, which frequently become chronic, associated with the ability of the bacteria to produce biofilm. Here, we report a relationship between the ability to produce chronic bovine mastitis and biofilm formation. We have classified bovine mastitis S. aureus isolates into three groups based on the presence of particular genetic elements required for biofilm formation: group 1 (ica(+) bap(+)), group 2 (ica(+), bap negative), and group 3 (ica negative, bap negative). Overall, animals naturally infected with group 1 and 2 isolates had a lower milk somatic cell count than those infected with isolates of group 3. In addition, Bap-positive isolates were significantly more able to colonize and persist in the bovine mammary gland in vivo and were less susceptible to antibiotic treatments when forming biofilms in vitro. Analysis of the structural bap gene revealed the existence of alternate forms of expression of the Bap protein in S. aureus isolates obtained under field conditions throughout the animal's life. The presence of anti-Bap antibodies in serum samples taken from animals with confirmed S. aureus infections indicated the production of Bap during infection. Furthermore, disruption of the ica operon in a bap-positive strain had no effect on in vitro biofilm formation, a finding which strongly suggested that Bap could compensate for the deficiency of the PIA/PNAG product (a biofilm matrix polysaccharide). Altogether, these results demonstrate that, in the bovine intramammary gland, the presence of Bap may facilitate a biofilm formation connected with the persistence of S. aureus.
MeSH Terms
Animals
Anti-Bacterial Agents/pharmacology
Bacterial Proteins/genetics,metabolism
Biofilms/growth & development
Cattle
Female
Mammary Glands, Animal/microbiology
Mastitis, Bovine/microbiology
Microbial Sensitivity Tests
Models, Animal
Sheep
Staphylococcal Infections/microbiology,veterinary
Staphylococcus aureus/drug effects,growth & development,pathogenicity
Tandem Repeat Sequences/genetics
Chemicals
Anti-Bacterial Agents
Bacterial Proteins
Bap protein, Staphylococcus aureus
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Cucarella Carme
Departament of Chemistry, Biochemistry, and Molecular Biology, Cardenal Herrera-CEU University, 46113 Moncada, Valencia, Spain.
Tormo M Angeles
Ubeda Carles
Trotonda M Pilar
Monzón Marta
Peris Critòfol
Amorena Beatriz
Lasa Iñigo
Penadés José R
References (29)
29 references, click to expand
-
Group B streptococci escape host immunity by deletion of tandem repeat elements of the alpha C protein.
Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4131-6
PMID: 8633028
-
Establishment of aging biofilms: possible mechanism of bacterial resistance to antimicrobial therapy.
Antimicrob Agents Chemother. 1992 Jul;36(7):1347-51
PMID: 1510427
-
The effects of early antibiotic treatment following diagnosis of mastitis detected by a change in the electrical conductivity of milk.
J Dairy Sci. 1997 May;80(5):859-63
PMID: 9178126
-
Molecular typing of Staphylococcus aureus based on PCR restriction fragment length polymorphism and DNA sequence analysis of the coagulase gene.
J Clin Microbiol. 1998 Apr;36(4):1083-9
PMID: 9542942
-
Infection-derived Enterococcus faecalis strains are enriched in esp, a gene encoding a novel surface protein.
Infect Immun. 1999 Jan;67(1):193-200
PMID: 9864215
-
Surface protein adhesins of Staphylococcus aureus.
Trends Microbiol. 1998 Dec;6(12):484-8
PMID: 10036727
-
Bacterial biofilms: a common cause of persistent infections.
Science. 1999 May 21;284(5418):1318-22
PMID: 10334980
-
Phase variation of slime production in Staphylococcus aureus: implications in colonization and virulence.
Infect Immun. 1993 Nov;61(11):4857-62
PMID: 8406887
-
Characterization of Tn917 insertion mutants of Staphylococcus epidermidis affected in biofilm formation.
Infect Immun. 1996 Jan;64(1):277-82
PMID: 8557351
-
Microbial biofilms.
Annu Rev Microbiol. 1995;49:711-45
PMID: 8561477
-
The intercellular adhesin involved in biofilm accumulation of Staphylococcus epidermidis is a linear beta-1,6-linked glucosaminoglycan: purification and structural analysis.
J Bacteriol. 1996 Jan;178(1):175-83
PMID: 8550413
-
Fibronectin binding protein and host cell tyrosine kinase are required for internalization of Staphylococcus aureus by epithelial cells.
Infect Immun. 1999 Sep;67(9):4673-8
PMID: 10456915
-
Antibiotic susceptibility assay for Staphylococcus aureus in biofilms developed in vitro.
J Antimicrob Chemother. 1999 Jul;44(1):43-55
PMID: 10459809
-
The intercellular adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation.
Infect Immun. 1999 Oct;67(10):5427-33
PMID: 10496925
-
Simplified scheme for routine identification of human Staphylococcus species.
J Clin Microbiol. 1975 Jan;1(1):82-8
PMID: 170303
-
Genetic analysis of functions involved in adhesion of Pseudomonas putida to seeds.
J Bacteriol. 2000 May;182(9):2363-9
PMID: 10762233
-
Role of biofilms in antimicrobial resistance.
ASAIO J. 2000 Nov-Dec;46(6):S47-52
PMID: 11110294
-
Bap, a Staphylococcus aureus surface protein involved in biofilm formation.
J Bacteriol. 2001 May;183(9):2888-96
PMID: 11292810
-
The enterococcal surface protein, Esp, is involved in Enterococcus faecalis biofilm formation.
Appl Environ Microbiol. 2001 Oct;67(10):4538-45
PMID: 11571153
-
Complete genome sequence of Salmonella enterica serovar Typhimurium LT2.
Nature. 2001 Oct 25;413(6858):852-6
PMID: 11677609
-
Synergy of different antibiotic combinations in biofilms of Staphylococcus epidermidis.
J Antimicrob Chemother. 2001 Dec;48(6):793-801
PMID: 11733463
-
Staphylococcus and biofilms.
Mol Microbiol. 2002 Mar;43(6):1367-78
PMID: 11952892
-
Expression of the biofilm-associated protein interferes with host protein receptors of Staphylococcus aureus and alters the infective process.
Infect Immun. 2002 Jun;70(6):3180-6
PMID: 12011013
-
Genome sequence survey identifies unique sequences and key virulence genes with unusual rates of amino Acid substitution in bovine Staphylococcus aureus.
Infect Immun. 2002 Jul;70(7):3978-81
PMID: 12065548
-
Immunochemical properties of the staphylococcal poly-N-acetylglucosamine surface polysaccharide.
Infect Immun. 2002 Aug;70(8):4433-40
PMID: 12117954
-
Genetic analysis of functions involved in the late stages of biofilm development in Burkholderia cepacia H111.
Mol Microbiol. 2002 Oct;46(2):411-26
PMID: 12406218
-
Biofilm testing of Staphylococcus epidermidis clinical isolates: low performance of vancomycin in relation to other antibiotics.
Diagn Microbiol Infect Dis. 2002 Dec;44(4):319-24
PMID: 12543535
-
Sip, an integrase protein with excision, circularization and integration activities, defines a new family of mobile Staphylococcus aureus pathogenicity islands.
Mol Microbiol. 2003 Jul;49(1):193-210
PMID: 12823821
-
Molecular basis of intercellular adhesion in the biofilm-forming Staphylococcus epidermidis.
Mol Microbiol. 1996 Jun;20(5):1083-91
PMID: 8809760