Skip to main content
SpringerLink
Log in

Molecular and phenotypic characterization of enterococci isolated from broiler flocks in Turkey

  • Regular Articles
  • Published:
Tropical Animal Health and Production Aims and scope Submit manuscript

Abstract

The aim of this study was to determine the antimicrobial resistance, resistance mechanisms implicated, and virulence genes (asa1, gelE, cylA, esp, and hyl) of Enterococcus spp. isolated from broiler flocks in Turkey. In addition, clonality of ampicillin and vancomycin-resistant enterococci was also investigated using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Out of 430 cloacal swab samples investigated, 336 (78.1%) Enterococcus spp. was isolated. The most frequently identified species were E. faecalis (87.8%), E. faecium (8.3%), E. durans (2.4%), E. casseliflavus (0.9%), and E. hirae (0.6%). The most common resistance was against tetracycline (81.3%), erythromycin (77.1%), ciprofloxacin (56.8%), and chloramphenicol (46.4%). Fifty (14.9%) isolates showed high-level gentamicin resistance (HLGL) phenotype. Ampicillin and vancomycin resistance were observed in 3.3% and 1.5% of the isolates, respectively. Two hundred eighty-three isolates were positive for the presence of virulence genes. Among the virulence genes tested, only gelE, asa1, esp, and cylA genes were detected. The most prevalent virulence gene was gelE (234, 69.6%), followed by asa1 (160, 47.6%), esp (37, 11%), and cylA (2, 0.6%). In conclusion, this study revealed that commensal enterococci from broiler flocks showed high rate of resistance to antimicrobials including clinically important antimicrobials for humans. The main underlying reason for high resistance could be attributed to the inappropriate and widespread use of antimicrobials. Therefore, there is an urgent need to develop control strategies to prevent the emergence and spread of antimicrobial resistance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Aarestrup, F.M., Agrees, Y., Gerner-Smith, P., Madsen, M., Jensen, L.B., 2000. Comparison of antimicrobial resistance phenotypes and resistance genes in Enterococcus faecalis and Enterococcus faecium from humans in the community, broilers and pigs in Denmark. Diagnostic Microbiology and Infectious Diseases, 37, 127–137.

    Article  CAS  Google Scholar 

  • Belhaj, M., Boutiba-Ben Boubaker, I., Ben Redjeb, S. and Bouchami, O., 2008. Molecular characterisation of high-level ampicillin-resistant Enterococcus faecium isolates from hospitalised patients in Tunis. International Journal of Antimicrobial Agents, 32, 284–286.

    Article  CAS  PubMed  Google Scholar 

  • Chajęcka-Wierzchowska, W., Zadernowska, A., Łaniewska-Trokenheim, L., 2017. Virulence factors of Enterococcus spp. presented in food. LWT-Food Science Technology, 75, 670–676.

    Google Scholar 

  • Cheng, A.C., Turnidge, J., Collignon, P., Looke, D., Barton, M., Gottlieb, T., 2012. Control of fluoroquinolone resistance through successful regulation, Australia. Emerging Infectious Disesases, 18, 1453–1460.

    Article  CAS  Google Scholar 

  • Chow, J.W., 2000. Aminoglycoside resistance in enterococci. Clinical Infectious Diseases, 31, 586–589.

    Article  CAS  PubMed  Google Scholar 

  • Clinical and Laboratory Standard Institute (CLSI): Performance Standards for Antimicrobial Susceptibility Testing; Nineteenth Informational Supplements. CLSI Document M100-S19. Wayne, PA, USA, 2012.

  • da Costa, P.M., Loureiro, L., Matos, A.J.F., 2013. Transfer of Multidrug-Resistant Bacteria between Intermingled Ecological Niches: The Interface between Humans, Animals and the Environment. International Journal of Environmental Research and Public Health, 10, 278–294.

    Article  PubMed  PubMed Central  Google Scholar 

  • Daikos, G.L., Bamias, G., Kattamis, C., Zervos, M.J., Chow, J.W., Christakis, G., Petrikkos, G., Triantafyllopoulou, P., Helen Alexandrou, H., Syriopoulou, V. 2003. Structures, locations, and transfer frequencies of genetic elements conferring high-level gentamicin resistance in Enterococcus faecalis isolates in Greece. Antimicrobial Agents and Chemotherapy, 47, 3950–3953

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • de Jong, A., Simjee, S., Garch, F. E., Moyaert, H., Rose, M., Youala, M., Dry, M., 2018. Antimicrobial susceptibility of enterococci recovered from healthy cattle, pigs and chickens in nine EU countries (EASSA Study) to critically important antibiotics. Veterinary Microbiology, 216, 168–175.

    Article  CAS  PubMed  Google Scholar 

  • del Campo, R., Tenorio, C., Rubio, C., Castillo, J., Torres, C., Gómez-Lus, R., 2000. Aminoglycoside-modifying enzymes in high-level streptomycin and gentamicin resistant Enterococcus spp. in Spain. International Journal Antimicrobial Agents, 15, 221–226.

    Article  Google Scholar 

  • Del Grosso, M., Caprioli, A., Chinzari, P., Fontana, M. C., Pezzotti, G., Manfrin, A., Giannatale, E.D., Goffredo, E., Pantosti, A., 2000. Detection and characterization of vancomycin-resistant enterococci in farm animals and raw meat products in Italy. Microbial Drug Resistance, 6, 313–318.

    Article  PubMed  Google Scholar 

  • Depardieu, F., Perichon, B., Courvalin, P., 2004. Detection of the van alphabet and identification of enterococci and staphylococci at the species level by multiplex PCR. Journal of Clinical Microbiology, 42, 5857–5860.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Diarra, M.S., Rempel, H., Champagne, J., Masson, L., Pritchard, J., Topp, E., 2010. Distribution of antimicrobial resistance and virulence genes in Enterococcus spp. and characterization of isolates from broiler chickens. Applied and Environmental Microbiology, 76, 8033–8043.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eaton, T.J., Gasson, M.J., 2001. Molecular screening of Enterococcus virulence determinants and potential for genetic exchange between food and medical isolates. Applied and Environmental Microbiology, 67, 1628–1635.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eisner, A., Feierl, G., Gorkiewicz, G., Dieber, F., Kessler, H. H., Marth, E., Köfer, J. 2005. High prevalence of VanA-type vancomycin-resistant Enterococci in Austrian poultry. Applied and Environmental Microbiology, 71, 6407–6409.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • el Amin, N.A., Jalal, S., Wretlind, B., 1999. Alterations in GyrA and ParC associated with fluoroquinolone resistance in Enterococcus faecium. Antimicrobial Agents Chemotherapy, 43, 947–949.

    Article  PubMed  Google Scholar 

  • Fisher, K., Phillips, C., 2009. The ecology, epidemiology and virulence of Enterococcus, Review. Microbiology, 155, 1749–1757.

    Article  CAS  PubMed  Google Scholar 

  • Hammerum, A.M., Lester, C.H., Heuer, O.E., 2010. Antimicrobial-resistant enterococci in animals and meat: a human health hazard?, Foodborne Pathogen Disease, 7, 1137–1146.

    Article  CAS  Google Scholar 

  • Hollenbeck, B.L., Rice, L.B., 2012. Intrinsic and acquired resistance mechanisms in enterococcus. Virulence, 3, 421–433.

    Article  PubMed  PubMed Central  Google Scholar 

  • Homan, W.L., Tribe, D., Poznanski, S., Li, M., Hogg, G., Spalburg, E., Van Embden, J.D., Willems, R.J., 2002. Multilocus sequence typing scheme for Enterococcus faecium. Journal of Clinical Microbiology, 40, 1963–1971.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hsieh, S.E., Hsu, L.L., Hsu, W.H., Chen, C.Y., Chen, H.J., Liao, C.T., 2006. Importance of amino acid alterations and expression of penicillin-binding protein 5 to ampicillin resistance of Enterococcus faecium in Taiwan. International Journal of Antimicrobial Agents, 28, 514–519.

    Article  CAS  PubMed  Google Scholar 

  • Jureen, R., Mohn, S.C., Harthug, S., Haarr, L., Langeland, N., 2004. Role of penicillin-binding protein 5 C-terminal amino acid substitutions in conferring ampicillin resistance in Norwegian clinical strains of Enterococcus faecium. APMIS, 112, 291–298.

    Article  CAS  PubMed  Google Scholar 

  • Layton, B.A., Walters, S.P., Lam, L.H., Boehm, A.B., 2010. Enterococcus species distribution among human and animal hosts using multiplex PCR. Journal of Applied Microbiology, 109, 539–547.

    CAS  PubMed  Google Scholar 

  • Leelaporn, A., Yodkamol, K., Waywa, D., Pattanachaiwit, S., 2008. A novel structure of Tn4001-truncated element, type V, in clinical enterococcal isolates and multiplex PCR for detecting aminoglycoside resistance genes. International Journal of Antimicrobial Agents, 31, 250–254.

    Article  CAS  PubMed  Google Scholar 

  • Liu, Y., Liu, K., Lai, J., Wu, C., Shen, J., Wang, Y., 2013. Prevalence and antimicrobial resistance of Enterococcus species of food animal origin from Beijing and Shandong Province, China. Journal of Applied Microbiology, 114, 555–563.

    Article  CAS  PubMed  Google Scholar 

  • Maasjost, J., Mühldorfer, K., Cortez de Jäckel, S., Hafez, H.M., 2015. Antimicrobial Susceptibility Patterns of Enterococcus faecalis and Enterococcus faecium Isolated from Poultry Flocks in Germany. Avian Diseases, 59, 143–148.

    Article  CAS  PubMed  Google Scholar 

  • Malhotra-Kumar, S., Lammens, C., Piessens, J., Goossens, H., 2005. Multiplex PCR for simultaneous detection of macrolide and tetracycline resistance determinants in streptococci. Antimicrobial Agents Chemotherapy, 49, 4798–4800.

    Article  CAS  PubMed  Google Scholar 

  • Marshall, B.M., Levy, S.B., 2011. Food animals and antimicrobials: impacts on human health. Clinical Microbiology Reviews, 24, 718–733.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mete, E., Kaleli, İ., Cevahir, N., Demir, M., Akkaya, Y., Kiriş Satılmış, Ö. 2017. Evaluation of virulence factors in enterococcus species. Bulletin of Microbiology, 51, 101–114.

    PubMed  Google Scholar 

  • Morrison, D., Woodford, N., Barrett, S.P., Sisson, P., Cookson, B.D., 1999. DNA Banding Pattern Polymorphism in Vancomycin-Resistant Enterococcus faecium and Criteria for Defining Strains. Journal of Clinical Microbiology, 37, 1084–1091.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ngbede, E.O., Raji, M.A., Kwanashie, C.N., Kwaga, J.K.P., 2017. Antimicrobial resistance and virulence profile of enterococci isolated from poultry and cattle sources in Nigeria. Tropical Animals and Health Production, 49, 451–458.

    Article  Google Scholar 

  • Nowakiewicz, A., Ziólkowska, G., Troscianczyk, A., Zieba, P., Gnat, S., 2017. Determination of resistance and virulence genes in Enterococcus faecalis and E. faecium strains isolated from poultry and their genotypic characterization by ADSRRS-fingerprinting. Poultry Science, 96, 986–996.

    CAS  PubMed  Google Scholar 

  • Obeng, A.S., Rickard, H., Ndi, O., Sexton, M., Barton, M., 2013. Comparison of antimicrobial resistance patterns in enterococci from intensive and free range chickens in Australia. Avian Pathology, 42, 45–54.

    Article  CAS  PubMed  Google Scholar 

  • Onodera Y, Okuda J, Tanaka M. and Sato, K., 2002. Inhibitory activities of quinolones against DNA gyrase and topoisomerase IV of Enterococcus faecalis. Antimicrobial Agents and Chemotherapy, 46, 1800–1804.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Palmer, K.L., Kos, V.N., Michael, S., Gilmore, M.S., 2010. Horizontal Gene Transfer and the Genomics of Enterococcal Antibiotic Resistance. Current Opinions in Microbiology, 13, 632–639.

    Article  CAS  Google Scholar 

  • Poeta, P., Costa, D., Igrejas, G., Sáenz, Y., Zarazaga, M., Rodrigues, J, Torres, C., 2007. Polymorphisms of the pbp5 gene and correlation with ampicillin resistance in Enterococcus faecium isolates of animal origin. Journal of Medical Microbiology, 56, 236–240.

    Article  CAS  PubMed  Google Scholar 

  • Rice, L.B., Bellais, S., Carias, L.L., Hutton-Thomas, R., Bonomo, R.A., Caspers, P., Page, M.G., Gutmann, L., 2004. Impact of specific pbp5 mutations on expression of beta-lactam resistance in Enterococcus faecium. Antimicrobial Agents and Chemotherapy, 48, 3028–3032.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rybkine, T., Mainardi, J.L., Sougakoff, W., Collatz, E., Gutmann, L., 1998. Penicillin-binding protein 5 sequence alterations in clinical isolates of Enterococcus faecium with different levels of beta-lactam resistance. Journal of Infectious Diseases, 178, 159–163.

    Article  CAS  PubMed  Google Scholar 

  • Saba Çopur, Ş., Şahin, F., Göçmen, J.S. 2016. Determination of virulence and multidrug resistance genes with polymerase chain reaction method in vancomycin-sensitive and -resistant enterococci isolated from clinical samples. Turk Journal of Medical Science, 46, 877–891.

    Article  CAS  Google Scholar 

  • Sava, I.G., Heikens, E., Kropec, A., Theilacker, C., Willems, R., Huebner, J., 2010. Enterococcal surface protein contributes to persistence in the host but is not a target of opsonic and protective antibodies in Enterococcus faecium infection. Journal of Medical Microbiology, 59, 1001–1004.

    Article  CAS  PubMed  Google Scholar 

  • Stępień-Pyśniak, D., Marek, A., Banach, T., Adaszek, Ł., Pyzik, E. and Wilczyński, J., Winiarczyk, S., 2016. Prevalence and antibiotic resistance of Enterococcus strains isolated from poultry. Acta Veterinaria Hungarica, 64, 148–63.

    Article  CAS  PubMed  Google Scholar 

  • Tejedor-Junco, M.T., Afonso-Rodríguez, O., Martín-Barrasa, J.L., González-Martín, M., 2005. Antimicrobial susceptibility of Enterococcus strains isolated from poultry faeces. Research in Veterinary Science, 78, 33–38.

    Article  CAS  PubMed  Google Scholar 

  • Turkish Ministry of Health (TMH) (2016): Summary Report of National Hospital Infections Surveying Network. 2016 Statistics, Ankara, Turkey. http//www.uhes.saglik.gov.tr Accessed 07 June 2018

  • Tzavaras, I., Siarkou, V.I., Zdragas, A., Kotzamanidis, C., Vafeas, G., Bourtzi-Hatzopoulou, E., Pournaras, S., Sofianou, D., 2012. Diversity of vanA-type vancomycin-resistant Enterococcus faecium isolated from broilers, poultry slaughterers and hospitalized humans in Greece. Journal of Antimicrobial Chemotherapy, 67, 1811–1818.

    Article  CAS  PubMed  Google Scholar 

  • Ünal, N., Aşkar, Ş. and Yıldırım, M. 2017. Antibiotic resistance profile of Enterococcus faecium and Enterococcus faecalis isolated from broiler cloacal samples. Turkish Journal of Veterinary and Animal Science, 41, 199–203.

    Article  CAS  Google Scholar 

  • Usui, M., Ozawa, S., Onozato, H., Kuge, R., Obata, Y., Uemae, T., Ngoc, P.T., Heriyanto, A., Chalemchaikit, T., Makita, K., Muramatsu, Y. and Tamura, Y., 2014. Antimicrobial susceptibility of indicator bacteria isolated from chickens in Southeast Asian countries (Vietnam, Indonesia and Thailand). Journal of Veterinary Medical Science, 76, 685–692.

    Article  CAS  PubMed  Google Scholar 

  • Vakulenko, S.B., Zervos, M.J., Donabedian, S.M., Lerner, S.A., Voskresenskiy A. M., Chow, J.W., 2003. Multiplex PCR fordetection of aminoglycoside resistance genes in enterococci. Antimicrobial Agents and Chemotherapy, 47, 1423–1426.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vankerckhoven, V., Van Autgaerden, T., Vael, C., Lammens, C., Chapelle, S., Rossi, R., Jabes, D., Goossens, H. 2004. Development of a multiplex PCR for the detection of asa1, gelE, cylA, esp and hyl genes in enterococci and survey for virulence determinants among European hospital isolates of Enterococcus faecium. Journal of Clinical Microbiology, 42, 4473–4479.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yilmaz, E.Ş., Aslantaş, Ö., Pehlivanlar Önen, S., Türkyilmaz, S., Kürekci, C., 2016. Prevalence, antimicrobial resistance and virulence traits in enterococci from food of animal origin in Turkey. LTW-Food Science Technology, 66, 20–26.

Download references

Funding

This work was supported by the Hatay Mustafa Kemal University Scientific Research Fund under Grant number of 15660.

Author information

Authors and Affiliations

  1. Department of Microbiology, Faculty of Veterinary Medicine, Hatay Mustafa Kemal University, 31060, Hatay, Turkey

    Özkan Aslantaş

Authors
  1. Özkan Aslantaş
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Özkan Aslantaş.

Ethics declarations

This study was approved by the Animal Ethical Committee of Mustafa Kemal University with decision number of 2015-9/2. Written informed consent was obtained from the owners of the broiler farms.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Aslantaş, Ö. Molecular and phenotypic characterization of enterococci isolated from broiler flocks in Turkey. Trop Anim Health Prod 51, 1073–1082 (2019). https://doi.org/10.1007/s11250-018-01784-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11250-018-01784-z

Keywords

Search

Navigation