Archives of Microbiology

, Volume 195, Issue 3, pp 203–209 | Cite as

Virulence determinants and biofilm production among Trueperella pyogenes recovered from abscesses of captive forest musk deer

  • Kelei Zhao
  • Yongqiang Tian
  • Bisong Yue
  • Hongning Wang
  • Xiuyue Zhang
Original Paper


Trueperella pyogenes (formerly Arcanobacterium) is commonly isolated from domesticated or wild ruminants as an opportunistic pathogen. To investigate the role of virulence determinants (VDs) and biofilm production in T. pyogenes isolates, a total of 36 T. pyogenes were collected from abscesses of forest musk deer in Miyaluo Farm (Sichuan Province, China). The prevalence of VDs and associations with clonal types, antibiotic resistance and biofilm production were analyzed by PCR and bioassay. Finally, T. pyogenes isolates were separated into three clonal types based on the DNA fingerprinting of BOX-PCR. Isolates with less VDs obtained from sick forest musk deer were mainly belonged to Type 1, and the isolates with robust VD repertoire obtained from dead forest musk deer were included in Type 3. Accordingly, resistant isolates exhibited significant lower virulence than susceptible ones. Majority of T. pyogenes isolates of this study were capable of producing a biofilm. However, no VDs presence and antibiotic resistance were statistically associated with biofilm production. In conclusion, the current study demonstrated that T. pyogenes was probably the primary pathogen of abscesses in the forest musk deer. Moreover, as an animal origin pathogen, the increasing resistance of T. pyogenes isolates could also associate with a decreased virulence.


Trueperella pyogenes Clonal type Virulence determinants Resistance Biofilm 



This work was supported by the National Basic Research Program of China (973 Project: 2012CB722207) and National Forestry Bureau of China (2130211). The authors would like to acknowledge Xuxin Li from Miyaluo Farm for collecting the samples. We thank Kiswendsida Ouedeaogo and the anonymous reviewers for editing the manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Bennett PM (2008) Plasmid encoded antibiotic resistance: acquisition and transfer of antibiotic resistance genes in bacteria. Br J Pharmacol 153:347–357CrossRefGoogle Scholar
  2. Brown ML, Aldrich HC, Gauthier JJ (1995) Relationship between glycocalyx and povidone-iodine resistance in Pseudomonas aeruginosa (ATCC 27853) biofilms. Appl Environ Microbiol 61(1):187–193PubMedGoogle Scholar
  3. Buntjer JB, Otsen M (2000) Cross checker provides computer-assisted marker interpretation. J Agric Genomics 4.
  4. Chen GL, Xu L, Yue BS, Zou FD (2007) Molecular cloning, characterizing of Interferon2 γ(IFN2γ) from forest musk deer (Moschus berezovskii) and its expression and purification in Escherichia coli. Sichuan J Zool 26:22–25Google Scholar
  5. CLSI (2009) Performance standards for antimicrobial susceptibility testing, 19th informational supplement, M100–S19. Clinical and Laboratory Standards Institute, WayneGoogle Scholar
  6. Crocetti GR, Hugenholtz P, Bond PL, Schuler A, Keller J, Jenkins D, Blackall LL (2000) Identification of polyphosphate-accumulating organisms and design of 16S rRNA-directed probes for their detection and quantitation. Appl Environ Microbiol 66:1175–1182PubMedCrossRefGoogle Scholar
  7. Davies D (2003) Understanding biofilm resistance to antibacterial agents. Nat Rev Drug Discov 2:114–122PubMedCrossRefGoogle Scholar
  8. Davies J, Davie D (2010) Origins and evolution of antibiotic resistance. Microbiol Mol Biol Rev 74:417–433PubMedCrossRefGoogle Scholar
  9. Esmay PA, Billington SJ, Link MA, Songer JG, Jost BH (2003) The Arcanobacterium pyogenes collagen binding protein, CbpA, promotes adhesion to host cells. Infect Immun 71:4368–4374PubMedCrossRefGoogle Scholar
  10. Guan TL, Zeng B, Peng QK, Yue BS, Zou FD (2009) Microsatellite analysis of the genetic structure of captive forest musk deer populations and its implication for conservation. Biochem Syst Ecol 37:166–173CrossRefGoogle Scholar
  11. Guha S, Goyal SP, Kashyap VK (2007) Molecular phylogeny of musk deer: a genomic view with mitochondrial 16S rRNA and cytochrome b gene. Mol Phylogenet Evol 42:585–597PubMedCrossRefGoogle Scholar
  12. Hoyle B, Costerton J (1991) Bacterial resistance to antibiotics: the role of biofilms. Prog Drug Res 37:91–105PubMedGoogle Scholar
  13. Johnson JR, Delavari P, Kuskowski M, Stell AL (2001) Phylogenetic distribution of extraintestinal virulence-associated traits in Escherichia coli. J Infect Dis 183:78–88PubMedCrossRefGoogle Scholar
  14. Johnson JR, Schee C, Kuskowski MA, Goessens WH, Belkum A (2002) Phylogenetic background and virulence profiles of fluoroquinolone-resistant clinical Escherichia coli isolates from the Netherlands. J Infect Dis 186:1852–1856PubMedCrossRefGoogle Scholar
  15. Johnson JR, Kuskowski MA, Owens K, Gajewski A, Winokur PL (2003) Phylogenetic origin and virulence genotype in relation to resistance to fluoroquinolones and/or extended-spectrum cephalosporins and cephamycins among Escherichia coli isolates from animals and humans. J Infect Dis 188:759–768PubMedCrossRefGoogle Scholar
  16. Jost BH, Billington SJ (2005) Arcanobacterium pyogenes: molecular pathogenesis of an animal opportunist. Antonie Van Leeuwenhoek Int J G 88:87–102CrossRefGoogle Scholar
  17. Jost BH, Songer JG, Billington SJ (1999) An Arcanobacterium (Actinomyces) pyogenes mutant deficient in production of the pore-forming cytolysin pyolysin has reduced virulence. Infect Immun 67:1723–1728PubMedGoogle Scholar
  18. Jost BH, Songer JG, Billington SJ (2002a) Identification of a second Arcanobacterium pyogenes neuraminidase and involvement of neuraminidase activity in host cell adhesion. Infect Immun 70:1106–1112PubMedCrossRefGoogle Scholar
  19. Jost BH, Post KW, Songer JG, Billington SJ (2002b) Isolation of Arcanobacterium pyogenes from the porcine gastric mucosa. Vet Res Commun 26:419–425PubMedCrossRefGoogle Scholar
  20. Lechtenberg KF, Nagaraja TG, Leipold HW, Chengappa MM (1988) Bacteriologic and histologic studies of hepatic abscesses in cattle. Am J Vet Res 49:58–62PubMedGoogle Scholar
  21. Louws FJ, Fulbright DW, Stephens CT, de Bruijn FJ (1994) Specific genomic fingerprints of phytopathogenic Xanthomonas and Pseudomonas pathovars and strains generated with repetitive sequences and PCR. Appl Environ Microbiol 60(Suppl. 7):2286–2295PubMedGoogle Scholar
  22. Martin B, Humbert O, Camara M, Guenzi E, Walker J, Mitchell T, Andrew P, Prudhomme M, Alloing G, Hakenbeck R, Morrison DA, Boulnois GJ, Claverys JP (1992) A highly conserved repeated DNA element located in the chromosome of Streptococcus pneumoniae. Nucleic Acids Res 20(Suppl. 13):3479–3483PubMedCrossRefGoogle Scholar
  23. Martinez-Martinez L, Fernandez F, Perea EJ (1999) Relationship between haemolysis production and resistance to fluoroquinolones among clinical isolates of Escherichia coli. J Antimicrob Chemother 43:277–279PubMedCrossRefGoogle Scholar
  24. Moreno E, Prats G, Sabaté M, Pérez T, Johnson JR, Andreu A (2006) Quinolone, fluoroquinolone and trimethoprim/sulfamethoxazole resistance in relation to virulence determinants and phylogenetic background among uropathogenic Escherichia coli. J Antimicrob Chemother 57:204–211PubMedCrossRefGoogle Scholar
  25. Narayanan S, Nagaraja TG, Wallace N, Staats J, Chengappa MM (1998) Biochemical and ribotypic comparison of Actinomyces pyogenes and Actinmyces pyogenes-like organisms from liver abscesses, rumianal wall, and ruminal contents of cattle. Am J Vet Res 59:271–276PubMedGoogle Scholar
  26. Natterman H, Horsch F (1977) The Corynebacterium pyogenes infection in cattle. 1. Incidence of the pathogen. Arch Exp Vet 31:405–413Google Scholar
  27. O’Toole GA, Kolter R (1998) Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol Microbiol 28:449–461PubMedCrossRefGoogle Scholar
  28. Pietrocola G, Valtulina V, Rindi S, Jost BH, Speziale P (2007) Functional and structural properties of CbpA, a collagen binding protein from Arcanobacterium pyogenes. Microbiology 153:3380–3389PubMedCrossRefGoogle Scholar
  29. Plamondon M, Martinez G, Raynal L, Touchette M, Valiquette L (2007) A fatal case of Arcanobacterium pyogenes endocarditis in a man with no identified animal contact: case report and review of the literature. Eur J Clin Microbiol Infect Dis 26:663–666PubMedCrossRefGoogle Scholar
  30. Pospiech A, Neumann B (1995) A versatile quick-prep of genomic DNA from Gram-positive bacteria. Trends Genet 11(Suppl. 6):217–218PubMedCrossRefGoogle Scholar
  31. Prüss BM, Besemann C, Denton A, Wolfe AJ (2006) A complex transcription network controls the early stages of biofilm development by Escherichia coli. J Bacteriol 188:3731–3739PubMedCrossRefGoogle Scholar
  32. Queen C, Ward ACS, Hunter DL (1994) Bacteria isolated from nasal and tonsillar samples of clinically healthy Rocky Mountain bighorn and domestic sheep. J Wildl Dis 30:1–7PubMedGoogle Scholar
  33. Silva E, Gaivao M, Leitao S, Jost BH, Carneiro C, Vilela CL, Lopes da Costa L, Mateus L (2008) Genomic characterization of Arcanobacterium pyogenes isolates recovered from the uterus of dairy cows with normal puerperium or clinical metritis. Vet Microbiol 132:111–118PubMedCrossRefGoogle Scholar
  34. Singh DV, Matte MH, Matte GR, Jiang S, Sabeena F, Shukla BN, Sanyal SC, Huq A, Colwell RR (2001) Molecular analysis of Vibrio cholerae O1, O139, non-O1, and non-O139 strains: clonal relationship between clinical and environmental isolates. Appl Environ Microbiol 67(Suppl. 2):910–921PubMedCrossRefGoogle Scholar
  35. Soto SM, Smithson A, Martinez JA, Horcajada JP, Mensa J, Vila J (2007) Biofilm formation in uropathogenic Escherichia coli strains: relationship with prostatitis, urovirulence factors and antimicrobial resistance. J Urol 177:365–368PubMedCrossRefGoogle Scholar
  36. Stewart PS, Costerton JW (2001) Antibiotic resistance of bacteria in biofilms. Lancet 358:135–138PubMedCrossRefGoogle Scholar
  37. Syrmis MW, O’Carrol MR, Sloots TP, Coulter C, Wainwright CE, Bell SC, Nissen MD (2004) Rapid genotyping of Pseudomonas aeruginosa isolates harboured by adult and paediatric patients with cystic fibrosis using repetitive-element-based PCR assays. J Med Microbiol 53:1089–1096PubMedCrossRefGoogle Scholar
  38. Vieira HL, Freire P, Arraiano CM (2004) Effect of Escherichia coli morphogene bolA on biofilms. Appl Environ Microbiol 70:5682–5684PubMedCrossRefGoogle Scholar
  39. Vila J, Simon K, Ruiz J, Horcajada JP, Velasco M, Barranco M, Moreno A, Mensa J (2002) Are quinolone-resistant uropathogenic Escherichia coli less virulent? J Infect Dis 186:1039–1042PubMedCrossRefGoogle Scholar
  40. Yassin AF, Hupfer H, Siering C, Schumann P (2011) Comparative chemotaxonomic and phylogenetic studies on the genus Arcanobacterium Collins et al. 1982 emend. Lehnen et al. 2006: proposal for Trueperella gen. nov. and emended description of the genus Arcanobacterium. Int J Syst Evol Microbiol 61:1265–1274PubMedCrossRefGoogle Scholar
  41. Zhao KL, Liu Y, Zhang XY, Palahati P, Wang HN, Yue BS (2011) Detection and characterization of antibiotic resistance genes in Arcanobacterium pyogenes strains from abscesses of forest musk deer. J Med Microbiol 60:1820–1826PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Kelei Zhao
    • 1
  • Yongqiang Tian
    • 2
  • Bisong Yue
    • 3
  • Hongning Wang
    • 3
  • Xiuyue Zhang
    • 1
  1. 1.Sichuan Key Laboratory of Conservation Biology on Endangered Wildlife, College of Life ScienceSichuan UniversityChengduPeople’s Republic of China
  2. 2.Laboratory of Chemical Biology, College of Chemical EngineeringSichuan UniversityChengduPeople’s Republic of China
  3. 3.Key Laboratory of Bioresources and Ecoenvironment (Ministry of Education), College of Life ScienceSichuan UniversityChengduPeople’s Republic of China

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