Staphylococcus aureus is a major pathogen associated to bovine mastitis and has the ability to form a slow-growing population termed the small colony variants (SCVs). From 20 samples of 5 chronic S. aureus cases, 1 SCV isolate (SCV102) was recovered simultaneously with 1 of 8 S. aureus isolates. SCV102 showed auxotrophy for thymidine and had a slow growth rate. Intracellular persistence in human mammary epithelial cells (HBL100cell line) monolayer revealed that SCV102 isolate had minimal cytopathological effects compared with its parent strains. SCV102 isolate and its parent strain S. aureus 101 indicate similar resistant pattern to four antibiotics. On the contrary, the minimal inhibitory concentrations values for chloramphenicol and sulfadimethoxine were much higher in SCV102 than that of S. aureus 101. To the best of our knowledge this is the first time the isolation of S. aureus SCV102 from a persistent bovine mastitis has been reported in Beijing (China). This study suggests that SCV102 isolate may be an important contributor to persistent bovine mastitis.
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References
1.
AtallaH., GylesC., JacobC.L., MoisanH., MalouinF., MallardB.2008. Characterization of a Staphylococcus aureus small colony variant (SCV) associated with persistent bovine mastitis. Foodborne Pathog. Dis., 5:785–799.
2.
BalwitJ.M., van LangeveldeP., VannJ.M., ProctorR.A.1994. Gentamicin-resistant menadione and hemin auxotrophic Staphylococcus aureus persist within cultured endothelial cells. J. Infect. Dis., 170:1033–1037.
3.
BarkemaH.W., SchukkenY.H., ZadoksR.N.2006. Invited Review: the role of cow, pathogen, and treatment regimen in the therapeutic success of bovine Staphylococcus aureus mastitis. J. Dairy Sci., 89:1877–1895.
4.
BesierS., SmacznyC., von MallinckrodtC., KrahlA., AckermannH., BradeV., WichelhausT.A.2007. Prevalence and clinical significance of Staphylococcus aureus small-colony variants in cystic fibrosis lung disease. J. Clin. Microbiol., 45:168–172.
5.
BesierS., ZanderJ., KahlB.C., KraiczyP., BradeV., WichelhausT.A.2008. The thymidine-dependent small-colony-variant phenotype is associated with hypermutability and antibiotic resistance in clinical Staphylococcus aureus isolates. Antimicrob. Agents Chemother., 52:2183–2189.
6.
BrakstadO.G., AasbakkK., MaelandJ.A.1992. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J. Clin. Microbiol., 30:1654–1660.
7.
ChambersH.F., MillerM.H.1987. Emergence of resistance to cephalothin and gentamicin during combination therapy for methicillin-resistant Staphylococcus aureus endocarditis in rabbits. J. Infect. Dis., 155:581–585.
8.
ChatterjeeI., BeckerP., GrundmeierM., BischoffM., SomervilleG.A., PetersG., SinhaB., HarraghyN., ProctorR.A., HerrmannM.2005. Staphylococcus aureus ClpC is required for stress resistance, aconitase activity, growth recovery, and death. J. Bacteriol., 187:4488–4496.
9.
ChatterjeeI., HerrmannM., ProctorR.A., PetersG., KahlB.C.2007. Enhanced post-stationary-phase survival of a clinical thymidine-dependent small-colony variant of Staphylococcus aureus results from lack of a functional tricarboxylic acid cycle. J. Bacteriol., 189:2936–2940.
10.
ChatterjeeI., KriegeskorteA., FischerA., DeiwickS., TheimannN., ProctorR.A., PetersG., HerrmannM., KahlB.C.2008. In vivo mutations of thymidylate synthase (encoded by thyA) are responsible for thymidine dependency in clinical small-colony variants of Staphylococcus aureus. J. Bacteriol., 190:834–842.
11.
ClementS., VaudauxP., FrancoisP., SchrenzelJ., HugglerE., KampfS., ChaponnierC., LewD., LacroixJ.-S.2005. Evidence of an intracellular reservoir in the nasal mucosa of patients with recurrent Staphylococcus aureus rhinosinusitis. J. Infect. Dis., 192:1023–1028.
12.
Clinical, Laboratory Standards Institute. 2006. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically. No. 2 Wayne, PA: Clinical and Laboratory Standards Institute. 26: Approved standard, 7thM7–A7.
13.
DevrieseL.1973. Hemin-dependent mutants isolated from methicillin-resistant Staphylococcus aureus strains. Antonie van Leeuwenhoek, 39:33–40.
14.
FosterT.J.2004. The Staphylococcus aureus “superbug”J. Clin. Invest., 114:1693–1696.
15.
GaoJ., FerreriM., YuF., LiuX., ChenL., SuJ., HanB.2011. Molecular types and antibiotic resistance of Staphylococcus aureus isolates from bovine mastitis in a single herd in China. Vet. J., 192:550–552.
16.
HardyK.J., OppenheimB.A., GossainS., GaoF., HawkeyP.M.2006. Use of variations in staphylococcal interspersed repeat units for molecular typing of methicillin-resistant Staphylococcus aureus strains. J. Clin. Microbiol., 44:271–273.
17.
HessD.J., Henry-StanleyM.J., EricksonE.A., WellsC.L.2003. Intracellular survival of Staphylococcus aureus within cultured enterocytes. J. Surg. Res., 114:42–49.
18.
KahlB., HerrmannM., EverdingA.S., KochH.G., BeckerK., HarmsE., ProctorR.A., PetersG.1998. Persistent infection with small colony variant strains of Staphylococcus aureus in patients with cystic fibrosis. J. Infect. Dis., 177:1023–1029.
19.
KahlB.C., BellingG., ReicheltR., HerrmannM., ProctorR.A., PetersG.2003. Thymidine-dependent small-colony variants of Staphylococcus aureus exhibit gross morphological and ultrastructural changes consistent with impaired cell separation. J. Clin. Microbiol., 41:410–413.
20.
KohlerC., von EiffC., PetersG., ProctorR.A., HeckerM., EngelmannS.2003. Physiological characterization of a heme-deficient mutant of Staphylococcus aureus by a proteomic approach. J. Bacteriol., 185:6928–6937.
21.
KuhnG., FrancioliP., BlancD.S.2006. Evidence for clonal evolution among highly polymorphic genes in methicillin-resistant Staphylococcus aureus. J. Bacteriol., 188:169–178.
22.
MelterO., RadojevcB.2010. Small colony variants of Staphylococcus aureus–review. Folia Microbiol., 55:548–558.
23.
MitchellG., GrondinG., BilodeauG., CantinA.M., MalouinF.2011. Infection of polarized airway epithelial cells by normal and small-colony variant strains of Staphylococcus aureus is increased in cells with abnormal cystic fibrosis transmembrane conductance regulator function and is influenced by NF-κB. Infect. Immun., 79:3541–3551.
24.
MoisanH., BrouilletteE., JacobC.L., Langlois-BeginP., MichaudS., MalouinF.2006. Transcription of virulence factors in Staphylococcus aureus small-colony variants isolated from cystic fibrosis patients is influenced by SigB. J. Bacteriol., 188:64–76.
25.
ProctorR.A., KahlB., von EiffC., VaudauxP.E., LewD.P., PetersG.1998. Staphylococcal small colony variants have novel mechanisms for antibiotic resistance. Clin. Infect. Dis., 27,Suppl 1:S68–S74.
26.
ProctorR.A., van LangeveldeP., KristjanssonM., MaslowJ.N., ArbeitR.D.1995. Persistent and relapsing infections associated with small-colony variants of Staphylococcus aureus. Clin. Infect. Dis., 20:95–102.
27.
ProctorR.A., von EiffC., KahlB.C., BeckerK., McNamaraP., HerrmannM., PetersG.2006. Small colony variants: a pathogenic form of bacteria that facilitates persistent and recurrent infections. Nat. Rev. Microbiol., 4:295–305.
28.
SabatA., Krzyszton-RussjanJ., StrzalkaW., FilipekR., KosowskaK., HryniewiczW., TravisJ., PotempaJ.2003. New method for typing Staphylococcus aureus strains: multiple-locus variable-number tandem repeat analysis of polymorphism and genetic relationships of clinical isolates. J. Clin. Microbiol., 41:1801–1804.
29.
SadowskaB., BonarA., von EiffC., ProctorR.A., ChmielaM., RudnickaW., RozalskaB.2002. Characteristics of Staphylococcus aureus, isolated from airways of cystic fibrosis patients, and their small colony variants. FEMS Immunol. Med. Microbiol., 32:191–197.
30.
SeggewissJ., BeckerK., KotteO., EisenacherM., YazdiM.R., FischerA., McNamaraP., Al LahamN., ProctorR., PetersG.2006. Reporter metabolite analysis of transcriptional profiles of a Staphylococcus aureus strain with normal phenotype and its isogenic hemB mutant displaying the small-colony-variant phenotype. J. Bacteriol., 188:7765–7777.
31.
SmithG.W., LymanR.L., AndersonK.L.2006. Efficacy of vaccination and antimicrobial treatment to eliminate chronic intramammary Staphylococcus aureus infections in dairy cattle. J. Am. Vet. Med. Assoc., 228:422–425.
32.
SommerhäuserJ., KloppertB., WolterW., ZschöckM., SobirajA., FailingK.2003. The epidemiology of Staphylococcus aureus infections from subclinical mastitis in dairy cows during a control programme. Vet. Microbiol., 96:91–102.
33.
SompolinskyD., CohenM., ZivG.1974. Epidemiological and biochemical studies on thiamine-less dwarf-colony variants of Staphylococcus aureus as etiological agents of bovine mastitis. Infect. Immun., 9:217–228.
34.
SompolinskyD., GluskinI., ZivG.1969. Pantothenate-requiring dwarf colony variants of Staphylococcus aureus as the etiological agent in bovine mastitis. J. Hyg. (Lond)., 67:511–516.
35.
TaoW., MallardB.2007. Differentially expressed genes associated with Staphylococcus aureus mastitis of Canadian Holstein cows. Vet. Immunol. Immunopathol., 120:201–211.
36.
TuchscherrL., HeitmannV., HussainM., ViemannD., RothJ., EiffC.v., PetersG., BeckerK., LöfflerB.2010. Staphylococcus aureus small-colony variants are adapted phenotypes for intracellular persistence. J. Infect. Dis., 202:1031–1040.
37.
von EiffC., BeckerK., MetzeD., LubritzG., HockmannJ., SchwarzT., PetersG.2001. Intracellular persistence of Staphylococcus aureus small-colony variants within keratinocytes: a cause for antibiotic treatment failure in a patient with Darier's disease. Clin. Infect. Dis., 32:1643–1647.
38.
von EiffC., BettinD., ProctorR.A., RolauffsB., LindnerN., WinkelmannW., PetersG.1997. Recovery of small colony variants of Staphylococcus aureus following gentamicin bead placement for osteomyelitis. Clin. Infect. Dis., 25:1250–1251.
39.
von EiffC., HeilmannC., ProctorR.A., WoltzC., PetersG., GotzF.1997. A site-directed Staphylococcus aureus hemB mutant is a small-colony variant which persists intracellularly. J. Bacteriol., 179:4706–4712.
40.
von EiffC., McNamaraP., BeckerK., BatesD., LeiX.H., ZimanM., BochnerB.R., PetersG., ProctorR.A.2006. Phenotype microarray profiling of Staphylococcus aureus menD and hemB mutants with the small-colony-variant phenotype. J. Bacteriol., 188:687–693.
41.
von EiffC., ProctorR.A., PetersG.2000. Staphylococcus aureus small colony variants: formation and clinical impact. Int. J. Clin. Pract. Suppl., 44–49.
42.
WiseR.I., SpinkW.W.1954. The influence of antibiotics on the origin of small colonies (G variants) of Micrococcus pyogenes var. aureus. J. Clin. Invest., 33:1611–1622.
