Molecular detection of extended spectrum ?-lactamase producing Klebsiella pneumoniae from wild deer




Abstract. Riwu KHP, Rantam FA, Effendi MH, Khairullah AR, Widodo A, Kurniawan SC, Ugbo EN. 2022. Molecular detection of extended spectrum ?-lactamase producing Klebsiella pneumoniae from wild deer. Biodiversitas 23: 4256-4262. Antimicrobial resistance (AMR) has become a severe threat worldwide and its remote cause remains unclear in most parts due to the global emergence of new resistance mechanisms by microorganisms. This research aimed to identify the presence of extended-spectrum ?-lactamase (ESBL) Klebsiella pneumoniae from wild deer feces in several places in East Java and East Nusa Tenggara. This study used 129 fresh feces samples collected from wild deer and analyzed for the presence of K. pneumoniae using standard microbiological techniques. An antibiotic sensitivity test was performed using Kirby-Bauer Disc Diffusion Method. The isolates were confirmed to the ESBL producer by double disk test and ESBL genes were identified by PCR. Nine samples (6.98%) were K. pneumoniae positive out of the 129 wild deer fresh feces. Among the nine K. pneumoniae isolates studied 3 (33.33%) were multidrug resistant, East Java (1 isolate) and East Nusa Tenggara (2 isolates). All the multidrug-resistant K. pneumoniae isolates were ESBL positive and also harbored blaTEM gene. The presence of MDR, and ESBL-producing K. pneumoniae isolates was confirmed in wild deer and this explained that wild deer should be considered a source of transmission of MDR and ESBL to public health.


Ali T, Rahman SU, Zhang L, Shahid M, Zhang S, Liu G, Gao J, Han B. 2016. ESBL-producing Escherichia coli from cows suffering mastitis in China contain clinical class 1 integrons with CTX-M linked to ISCR1. Frontiers in Cellular and Infection Microbiology. DOI: 10.3389/fmicb.2016.01931.
Allen SE, Boerlin P, Janecko N, Lumsden JS, Barker IK, Pearl DL, Jardine C. 2011. Antimicrobial resistance in generic Escherichia coli isolates from wild small mammals living in swine farm, residential, landfill, and natural environments in southern Ontario, Canada. Appl Environ Microbiol , 77(3), 882-888.
Barco L, Barrucci F, Olsen JE, Ricci A. 2013. Salmonella source attribution based on microbial subtyping. International Journal of Food Microbiology. 163.193-203.
Bender JB, Shulman SA. 2004. Reports of zoonotic disease outbreaks associated with animal exhibits and availability of recommendations for preventing zoonotic disease transmission from animals to people in such settings. J Am Vet Med Assoc , 224(7), 1105-1109.
Bradley ME, Scoular SK. 2019. Metastatic Klebsiella pneumoniae Invasive liver abscess syndrome in Denver, Colorado. J Pharm Pract. DOI: 10.1177/0897190019882867.
Bréchet C, Plantin J, Sauget M, Thouverez M, Talon D, Cholley P, et al. 2014. Wastewater treatment plants release large amounts of extended-spectrum ?-lactamase–producing Escherichia coli into the environment. Clin. Infect. Dis. DOI: 10.1093/cid/ciu190.
CDC. 2015.CDC works 24/7 to protect US from health, safety and security threats. CDC, Atlanta, GA.
CLSI. 2017, M100 Performance Standards for Antimicrobial. 27th ed. Clinical and Laboratory Standards Institute, USA.
Correa L, Martino MD, Siqueira I, Pasternak J, Gales AC, Silva CV, Camargo TZS, Scherer PF, Marra AR. 2013. A hospital-based matched case control study to identify clinical outcome and risk factors associated with carbapenem-resistant Klebsiella pneumoniae infection. BMC Infect Dis. 13:80. DOI: 10.1186/1471-2334-13-80.
Dallenne C, Da Costa A, Decre D, Favier C, Arlet G. 2010. Development of a set of multiplex PCR assays for the detection of genes encoding important beta-lactamases in Enterobacteriaceae. J. Antimicrob. Chemother.65:490–495. DOI: 10.1093/jac/dkp498.
de Been M, Lanza VF, de Toro M, Scharriga J, Dohmen W, Du Y, et al. 2014. Dissemination of cephalosporin resistance genes between Escherichia coli strains from farm animals and humans by specific plasmid lineages. PLoS Genetics. DOI: 10.1371/journal.pgen.1004776.
Dierikx CM, Van der Goot J, Fabri T, Van EssenZandbergen A, Smith H, Mevius D. 2012. Extended Spectrum-beta-lactamase- and AmpC-beta lactamase-producing Escherichia coli in Dutch broilers and broiler farmers. Journal of Antimicrobial Chemotherapy. DOI: 10.1093/jac/dks349
Dierikx CM, van Essen-Zandbergen A, Veldman K, Smith H, Mevius D. 2010. Increased detection of extended spectrum beta-lactamase producing Salmonella enterica and Escherichia coli isolates from poultry. Veterinary Microbiology. DOI: 10.1016/j.vetmic.2010.03.019.
Domingues AR, Pires SM, Halasa T, Halt T. 2012. Source attribution of human salmonellosis using a meta analysis of case-control studies of sporadic infections. Epidemiology and Infection. DOI: 10.1017/S0950268811002172
Domingues AR, Pires SM, Halasa T, Halt T. 2012. Source attribution of human campylobacteriosis using a meta-analysis of case-control studies of sporadic infections. Epidemiology and Infection. DOI: 10.1017/S0950268811002676.
Dsouza R, Pinto NA, Hwang I, Cho Y, Yong D, Choi J, Lee K, Chong Y. 2017. Panel strain of Klebsiella pneumonia for betalactam antibiotic evaluation their phenotypic and genotypic characterization.; PeerJ. DOI: 10.7717/peerj.2896.
Eckmanns T, Richter D, Feig M. 2014. MRSA and ESBL in outpatient: development from 2008 up to 2012 and socio demographic differences. Berl Munch Tierarztl Wochenschr 127:399–402.
Effendi M H. Bintari I G, Aksono E B, Hermawan I P. Detection of blaTEM Gene of Klebsiella pneumoniae Isolated from swab of food-producing animals in East Java. Trop. Anim. Sci. J. 2018; 41:174–178.
Effendi M.H, Harijani N, Budiarto, Triningtya N.P, Tyasningsih W. and Plumeriastuti H. Prevalence of Pathogenic Escherichia Coli Isolated from Subclinical Mastitis in East Java Province, Indonesia. Indian Vet. J. 2019; 96 (03): 22 – 25.
EFSA (European Food Safety Authority). 2011. Scientific opinion on the public health risks of bacterial strains producing extended-spectrum beta-lactamases and/or AmpC beta-lactamases in food and foodproducing animals. EFSA Journal. DOI:10.2903/j.efsa.2011.2322.
Evers EG, Van der Vels-Klerx HJ, Nauta MJ, Schjiven J, Havelaar AH. 2008. Campylobacter source attribution by exposure assessment. International Journal of Risk Assessment and Management. DOI:10.1504/IJRAM.2008.016151
Fair RJ, Tor Y. 2014. Antibiotics and bacterial resistance in the 21st Century. Perspect.Med.Chem. DOI: 10.4137/PMC.S14459
Founou LL, Founou RC, Essack SY. 2016. Antibiotic resistance in the food chain: a developing country perspective. Front. Microbiol. DOI: 10.3389/fmicb.2016.01881.
Geser N, Stephan R, Korczak BM, Beutin L, Hachler H. 2012. Molecular identification of extended-spectrum?lactamase genes from Enterobacteriaceae isolated from healthy human carriers in Switzerland. Antimicrobial Agents and Chemotherapy DOI: 10.1128/AAC.05539-11.
Guo Y, Zhou H, Qin L, Pang Z, Qin T, Ren H, Pan Z, Zhou J. 2016. Frequency, antimicrobial resistance and genetic diversity of Klebsiella pneumoniae in food samples. PLoS One. DOI: 10.1371/journal.pone.0153561
Hartmann A, Locatelli A, Amoureux L, Depret G, Jolivet C, Gueneau E, et al. 2012. Occurrence of CTX-M producing Escherichia coli in soils, cattle, and farm environment in France (Burgundy region). Frontiers in Microbiology. DOI: 10.3389/fmicb.2012.00083.
Hasibuan M, Suryanto D, Kusumawati RL. Phenotypic and molecular detection of blaCTX-M gene Extended Spectrum Beta-Lactamases in Escherichia coli and Klebsiella pneumoniae of North Sumatera isolates. 2018. IOP Conference Series: Earth and Environmental Science. DOI: 10.1088/1755-1315/130/1/012032.
Hertl JA, Schukken YH, Welcome FL, Tauer LW, Grohn YT. 2014. Pathogen- specific effects on milk yield in repeated clinical mastitis episodes in Holstein dairy cows. J Dairy Sci. DOI: 10.3168/jds.2013-7266.
Jean SS, Hsueh PR. 2011. High burden of antimicrobial resistance in Asia. Int J Antimicrob Agents, 37(4), 291-295.
Kikuchi N, Blakeslee JR, Hiramune T . 1995. Plasmid profiles of Klebsiella pneumoniae isolated from horses. J Vet Med Sci. DOI: 10.1292/jvms.57.113.
Kristianingtyas L, Effendi, MH, Tyasningsih W, Kurniawan F. 2020. Genetic Identification of blactx-M Gene and blatem Gene on Extended Spectrum Beta Lactamase (ESBL) Producing Escherichia Coli from Dogs. Indian Vet. J. 97 (01): 17 – 21.
Kusumaningrum HD, Handayani L, Novrianti R. 2016. Partial sequencing of 16S rRNA gene of selected Staphylococcus aureus isolates and its antibiotic resistance. Med. Pet. DOI: 10.5398/medpet.2016.39.2.67.
Laupland KB, Church DL, Vidakovich J, Mucenski M, Pitout JDD. 2008. Community-onset extended-spectrum beta-lactamase (ESBL) producing Escherichia coli: importance of international travel. Journal of Infection. DOI: 10.1016/j.jinf.2008.09.034.
Laxminarayan R, Duse A, Wattal C, Zaidi AKM, Wertheim HFL, Sumpradit N, et al. 2013. Antibiotic resistance-the need for global solutions. Lancet Infect. Dis 13, 1057-1098.
Lestari NW, Budiharjo A, Pangastuti A. 2016. Aerobic heterotrophic bacteria from the digestive tract of eel (Anguilla bicolor bicolor) and their potential as probiotics. Biotechnology. DOI: 10.13057/biotek/c130102.
Leverstein-van Hall MA, Dierikx CM, Cohen Stuart J, Voets GM, Van Den Munkchof MP, Van EssenZandbergen A, et al. 2011. Dutch patients, retail chicken meat and poultry share the same ESBL genes, plasmids and strains. Clinical Microbiology and Infection 17: 873–880.
Ma KL, Wang CX. 2013. Analysis of the spectrum and antibiotic resistance of uropathogens in vitro: Result based on a retrospective study from a tertiary hospital. AMJ Infect Control; 41:601-606.
Masruroh CA, Sudarwanto MB, Latif H. 2016. Incidence rate of Escherichia coli producing Extended Spectrum ?eta-Lactamase isolated from broiler faeces in Bogor. J Sains Veteriner. 2016; 34: 42–49.
Ningrum SG, Soejoedono RD, Latif H, Arnafia W, Wibawan IWT. 2016. Prevalence and characterization of shiga toxin-producing Escherichia coli isolated from slaughtered qurban animal in Jakarta Province. Med. Pet. 39: 90-94. DOI: 10.5398/medpet.2016.39.2.90.
Okonko IO, Soleye FA, Amusan TA, Ogun AA, Ogunnusi TA, Ejembi J, et al. 2009. Incidence of multi-drug resistance (MDR) organisms in Abeokuta, Southwestern Nigeria. Global J Pharmacol. 3: 69-80.
Overdevest I, Willemsen I, Rijnsburger M, Eustace A, Xu L, Hawkey P, et al. 2011. Extended spectrum beta lactamase genes of Escherichia coli in chicken meat and humans, The Netherlands. Emerging Infectious Diseases 17:1216-1222.
Paulin-Curlee GG, Sreevatsan S, Singer RS, Isaacson R, Reneau J, Bey R, Foster D. 2008. Molecular subtyping of mastitis-associated Klebsiella pneumoniae isolates shows high levels of diversity within and between dairy herds. J Dairy Sci. DOI: 10.3168/jds.2007-0479.
Perron G, Quessy G, S, Bell G. 2008. A reservoir of drug-resistant pathogenic bacteria in asymptomatic hosts. PLoS One. DOI: 10.1371/journal.pone.0003749.
Pires SM, Evers EG, van Pelt W, Ayers T, Scallan E, Angulo FJ, et al.2009. Attributing the human disease burden of foodborne infections to specific sources. Foodborne Pathogens and Disease. DOI:10.1089/fpd.2008.0208.
Putra AR. Effendi MH, Koesdarto S, Suwarno S, Tyasningsih W, Estoepangestie AT. Detection of the extended spectrum ?-lactamase produced by Escherichia coli from dairy cows by using the Vitek-2 method in Tulungagung regency, Indonesia. 2020. Iraqi Journal of Veterinary Sciences. DOI:10.33899/ijvs.2019.125707.1134.
Rahmahani J, Salamah, Mufasirin, Tyasningsih W, and Effendi MH. 2020. Antimicrobial Resistance Profile of Escherichia coli From Cloacal Swab of Domestic Chicken in Surabaya Traditional Market. Biochem Cell Arch 20 (1): 2993-2997. DOI: 10.35124/bca.2020.20. S1.2993
Remschmidt C, Schneider S, Meyer E, Schroeren-Boersch B, Gastmeier P, Schwab F. 2017. Surveillance of antibiotic use and resistance in intensive care units (SARI). Dtsch Arztebl Int DOI: 10.3238/arztebl.2017.0858.
Riwu KHP, Effendi MH, Rantam FA. 2020. A review of extended-spectrum?-Lactamase (ESBL) producing Klebsiella pneumoniae and Multidrug-Resistant (MDR) on companion animals.SystRev Pharm 11(7):270-277. DOI:10.31838/srp.2020.7.43.
RKI, Epidemiologisches Bulletin. Robert Koch Institut 2013. no 19: pp 167–171.
Sagita, D., L. Azizah, & Y. Sari. 2015. Identification of bacteria and antibiotic sensivity test of pus of post surgical wound infection in Jambi Public Hospital in the period August-October 2014. Sains Farmasi 1: 6-7.
Saishu N, Ozaki H, Murase T. 2014. CTX-M-type extended-spectrum beta- lactamase-producing Klebsiella pneumoniae isolated from cases of bovine mastitis in Japan. J Vet Med Sci. DOI: 10.1292/jvms.13-0120.
Santajit S, Indrawattana N. 2016. Mechanisms of antimicrobial resistance in Eskape pathogens. Biomed Res Int. DOI: 10.1155/2016/2475067.
Savira M. 2014. The validity of the conventional method is modified to the conventional method and the ESBL chromidTM for the detection of Extended-Spectrum Beta-Lactamases producing bacteria. JIK. 2: 81– 90. 15.
Smet A, Martel A, Persoons D, Dewuif J, Heyndrickx M, Cloeckaert A. 2009. Comparative analysis of extended spectrum beta-lactamase-carrying plasmids from different members of Enterobacteriaceae isolated from poultry, pigs and humans: evidence for a shared beta-lactam resistance gene pool. Journal of Antimicrobial Chemotherapy. DOI: 10.1093/jac/dkp101.
Smet A, Rasschaert G, Martel A, Persoons D. 2011. In situ ESBL conjugation from avian to human Escherichia coli during cefotaxime administration. Journal of Applied Microbiology. DOI: 10.1111/j.1365-2672.2010.04907.x.
Vinué L, Sáenz Y, Martínez S, Somalo M, Moreno Ma, Torres C, et al. 2009. Prevalence and diversity of extended spectrum ?lactamases in faecal Escherichia coli isolates from healthy humans in Spain. Clinical Microbiology and Infection 15:954-956.
Wibisono, F. J., Sumiarto, B., Untari, T., Effendi, M.H., Permatasari, D.A., and Witaningrum, A.M. 2020. Short Communication: Pattern of Antibiotic Resistance on Extended-Spectrum Beta-Lactamases Genes Producing Escherichia coli on Laying Hens in Blitar, Indonesia. Biodiversitas, 21 (10): 4631-4635
Widodo, A., Effendi, M.H., and Khairullah, A.R. (2020). Extended-Spectrum Beta-Lactamase (ESBL)-Producing Escherichia coli from Livestock. Sys Rev Pharm, 11 (7): 382-392.
Xue Wang, Qian Kang, Jianan Zhao, et al. Characteristics and epidemiology of extended-spectrum ?-lactamase-producing Multidrug-Resistant Klebsiella pneumoniae from Red Kangaroo,China. Fmicb: 2020. 560474.
Yang X, Wai-Chi CE, Zhang R, Chen S. (2019). A conjugative plasmid that augments virulence in Klebsiella pneumoniae .Nat Microbiol , 4(12), 2039-2043.

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