Characterization of Faecal Enterococci from Wild Birds in Turkey and Its Importance in Antimicrobial Resistance

Wild birds enterococci antibiotic resistance vancomycin resistance virulence factors

This research aimed to investigate the diversity of faecal enterococci isolated from wild birds, to detecttheir antibiotic resistance patterns and to determine their distribution of genes related to vancomycin resistance. Additionally, to investigate their virulence factors that are important in the development of the disease. One hundred seven cloacal/rectal samples were inoculated onto Enterococcus Agar, and presumptive colonies were identified and confirmed by PCR. Multiplex PCR assays were used to screen vanA, vanB, vanC1 and vanC2/3. The virulence-related genes; ace, gelE, efa and agg were determined by PCR. Among the 103 enterococci, 62 E.faecalis, 23 E.faecium 3 E.gallinarum, 2 E.durans, 1 E.casseliflavus and 12 Enterococcus spp. were identified. Of the 103 enterococci, 26 were found to be resistant against to three or more antibiotics. The highest percentages were detected for chloramphenicol (52%), tetracycline (33%) and erythromycin (30%). Two E.gallinarum isolates were harboring three virulence factors, and one isolate was carrying a single virulence factor. There is no virulence factor in the E.casseliflavus isolate. Also, vanA and vanB genes were not found. Forty-two of 103 enterococci were harboring virulence factors, more frequently in E.faecalis. Forty-two enterococci carried efa A, 31 isolates carried gel E, and ace was found in 18 isolates. Virulence gene agg was not detected. When the results of the study were evaluated in general, multiple drug resistance was described as 25%. Considering the risk of polluting the water resources of wild animals, it is suggested that the continuity of this type of epidemiological study in wildlife animals is necessary. In conclusion, the wild birds may act as substantial reservoirs carrying antimicrobial resistance among enterococci and estimate the potential risk for man, pets and farm animals.

Article Details
  • Section
  • Research Articles
Download data is not yet available.
Aarestrup FM (2005) Veterinary drug usage and antimicrobial resistance in bacteria of animal origin. Basic & clinical pharmacology & toxicology 96(4): 271-281.
Akgül Ö, Gülhan T, Güdücüoğlu H (2016) Tavuk ve martı kökenli enterokok türlerinin antibiyotik dirençliliğinin fenotipik ve genotipik analizi. Vet J Ankara Univ 63: 235-244.
Bagcigil AF, Koenhemsi L, Çelik B, Metiner K, Or ME, Ak S (2016) Examination of vancomycin-resistant Enterococci (VRE) isolated from canine and feline rectal swabs. J Fac Vet Med Istanbul Univ 42(2): 111-116.
Blanco G, Lemus JA, Grande J (2006) Faecal bacteria associated with different diets of wintering red kites: influence of livestock carcass dumps in microflora alterationand pathogen acquisition. J Appl Ecol 43: 990-998.
Celik B, Bagcigil AF, Koenhemsi L, Adiguzel MC, Or ME, Ak S (2017) Determination of Ampicillin Resistant Enterococci (ARE) Isolated From Canine and Feline Rectal Swabs. J Fac Vet Med Istanbul Univ 43 (1): 1-6.
Cetinkaya Y, Falk P, Mayhall CG (2000)Vancomycin-resistant enterococci. Clin Microbiol Rev 13(4) : 686-707.
Clinical and Laboratory Standard Institute (2016) Performance Standards for Antimicrobial Susceptibility Testing; 22nd Informational Supplement M100-A22. Wayne, PA: Clinical and Laboratory Standards Institute.
Creti R, Imperi M, Bertuccini L, Fabretti F, Orefici G, Di Rosa R, Baldassarri L (2004) Survey for virulence determinants among Enterococcus faecalis isolated from different sources. J Med Microbiol 53: 13-20.
DevrieseL, Pot B, Kersters K, Lauwers S, Haesebrouck F (1996) Acidification of methyl-a-D-glucopyranoside: A useful test to differentiate Enterococcus casseliflavus and Enterococcus gallinarum from Enterococcus faecium species group and from Enterococcus faecalis. J Clin Microbiol 34: 2607-2608.
Eaton TJ, Gasson MJ (2001) Molecular screening of Enterococcus virulence determinants and potential for genetic exchange between food and medical isolates. Appl Environ Microbiol 67: 1628-1635.
Han D, Unno T, Jang J, Lim K, Lee SN, Ko G, Hur HG (2011) The occurrence of virulence traits among high-level aminoglycosides resistant Enterococcus isolates obtained from faeces of humans, animals, and birds in South Korea. Int J Food Microbiol 144(3): 387-392.
Jánošková A, Kme V (2004) Vancomycin resistance genes in Enterococcus spp. strains isolated from alpine accentor and chamois. Acta Vet Brno 73(2): 211-214.
Kariyama R, Mitsuhata R, Chow JW, Clewell DB, Kumon H (2000)Simple and reliable Multiplex-PCR assay for surveillance isolates of vancomycin-resistant enterococci. J Clin Microbiol 38: 3092-3095.
Ke D, Picard FJ, Martineau F, Ménard C, Roy PH, Ouellette M, Bergeron MG (1999) Development of a PCR assay for rapid detection of Enterococci. J Clin Microbiol 37(11): 3497-3503.
Klibi N, Amor IB, Rahmouni M, Dziri R, Douja G, Said LB, Torres C (2015) Diversity of species and antibiotic resistance among fecal enterococci from wild birds in Tunisia. Detection of vanA-containing Enterococcus faecium isolates. Eur J Wildl Res 61(2): 319-323.
Lopez M, Tenorio C, Torres C (2011) Study of vancomycin resistance in faecal enterococci from healthy humans and dogs in Spain a decade after the avoparcin ban in Europe. Zoonoses Public Health 60:160-167.
Lozano C, Gonzalez-Barrio D, Camacho MC, Lima-Barbero JF, de la Puente J, Höfle U, Torres C (2016) Characterization of fecal vancomycin-resistant enterococci with acquired and intrinsic resistance mechanisms in wild animals, Spain. Microbial ecology 72(4): 813-820.
Mannu L, Paba A, Daga E, Comunian, R, Zanetti S, Duprè I, Sechi LA (2003) Comparison of the incidence of virulence determinants and antibiotic resistance between Enterococcus faecium strains of dairy, animal and clinical origin. Int J of Food Microbiol 88(2): 291-304.
Marrow J, Whittington JK, Mitchell M, Hoyer LL, Maddox C (2009) Prevalence and antibiotic-resistance characteristics of Enterococcus spp. Isolated from free-living and captive raptors in central Illinois. J Wildlife Dis 45(2): 302-313.
OIE list of antimicrobial agents of veterinary importance. Paris. World Organisation for Animal Health (2015) ( (Accessed 10 January 2019)
Oravcova V, Zurek L, Townsend A, Clark AB, Ellis JC, Cizek A, Literak I (2014) American crows as carriers of vancomycin‐resistant enterococci with vanA gene. Environ Int 16(4): 939-949.
Oravcová V, Peixe L, Coque TM, Novais C, Francia MV, Literák I, Freitas AR (2018) Wild corvid birds colonized with vancomycin-resistant Enterococcus faecium of human origin harbour epidemic vanA plasmids. Environ Int 118: 125-133.
Poeta P, Costa D, Sáenz Y, Klibi N, Ruiz‐Larrea F, Rodrigues J, Torres C (2005) Characterization of antibiotic resistance genes and virulence factors in faecal enterococci of wild animals in Portugal. J Vet Med 52(9):396-402.
Radhouani H, Pinto L, Coelho C, Sargo R, Araujo C, Lopez M, Torres C, Igrejas G, Poeta P (2010) MLST and a genetic study of antibiotic resistance and virulence factors in vanA-containing Enterococcus from buzzards (Buteo buteo). Lett Appl Microbiol 50: 537-541.
Radhouani H, Poeta P, Goncalves A, Pacheco R, Sargo R, Igrejas G (2012) Wild birds as biological indicators of environmental pollution: antimicrobial resistance patterns of Escherichia coli and enterococci isolated from common buzzards (Buteo buteo). J Med Microbiol 61(6): 837-843.
Radhouani H, Silva N, Poeta P, Torres C, Correia S, Igrejas G (2014) Potential impact of antimicrobial resistance in wildlife, environment and human health. Frontiers in microbial 5: 23.
Radimersky T, Frolkova P, Janoszowska D, Dolejska M, Svec P, Roubalova E, Literak I (2010) Antibiotic resistance in faecal bacteria (Escherichia coli, Enterococcus spp.) in feral pigeons. J App Microbiol 109(5): 1687-1695.
Santos T, Silva N, Igrejas G, Rodrigues P, Micael J, Rodrigues T, Cunha R (2013) Dissemination of antibiotic-resistant Enterococcus spp. and Escherichia coli from wild birds of Azores Archipelago. Anaerobe 24: 25-31.
Sellin M, Palmgren H, Broman T, Bergström S, Olsen B (2000) Involving ornithologists in the surveillance of vancomycin-resistant enterococci. Emerg Infect Dis 6(1): 87.
Semedo T, Santos MA, Lopes MF, Marques JJF, Crespo MT, Tenreiro R (2003)Virulence factors in food, clinical and reference enterococci: a common trait in the genus? Syst Appl Microbiol 26(1): 13-22.
Silva N, Igrejas G, Rodrigues P, Rodrigues T, Gonçalves A, Felgar AC, Poeta P (2011) Molecular characterization of vancomycin-resistant enterococci and extended-spectrum β-lactamase-containing Escherichia coli isolates in wild birds from the Azores Archipelago. Avian Pathol 40(5): 473-479.
Splichalova P, Svec P, Ghosh A, Zurek L, Oravcova V, Radimersky T, Literak I (2015) Prevalence, diversity and characterization of enterococci from three coraciiform birds. Antonie Van Leeuwenhoek 107(5): 1281-1289.
Vittecoq M, Godreuil S, Prugnolle F, Durand P, Brazier L, Renaud N, Thomas F (2016) Antimicrobial resistance in wildlife. J App Ecol 53(2): 519-529.
Werner G (2012) Current Trends of Emergence and Spread of Vancomycin-Resistant Enterococci. In: Dr Marina Pana (Ed.) ed. Antibiotic-Resistant Bacteria - A Continuous Challenge in the New Millennium: InTech 303-355.
World Health Organization (2017) Critically important antimicrobials for human medicine: ranking of antimicrobial agents for risk management of antimicrobial resistance due to non-human use. (Accessed 10 January 2019)
Yahia HB, Chairat S, Hamdi N, Gharsa H, Sallem RB, Ceballos S, Slama KB (2018) Antimicrobial resistance and genetic lineages of faecal enterococci of wild birds: Emergence of vanA and vanB2 harbouring Enterococcus faecalis. Int J of Antimicro Ag 52: 936-941.