Restricted accessResearch articleFirst published online 2018-03
Routine Prophylactic Antimicrobial Use Is Associated with Increased Phenotypic and Genotypic Resistance in Commensal Escherichia coli Isolates Recovered from Healthy Fattening Pigs on Farms in Thailand
This study examined antimicrobial resistance (AMR) profiles in commensal Escherichia coli derived from healthy fattening pigs in Thai farms that used prophylactic antimicrobials (in-feed tiamulin fumarate and amoxicillin) [PAs], therapeutic antimicrobials (injectable enrofloxacin or gentamicin) [TAs], or no antimicrobials [NAs]. Commensal E. coli were used as a proxy for overall AMR on the farms. There was a high level of multidrug resistance in all three categories of farm, with isolates showing resistance to β-lactams (amoxicillin, ampicillin, and piperacillin) and tetracyclines (tetracycline), and commonly possessing tetA, blaTEM, and plasmid replicons FIB and F. On the other hand, isolates with an extended-spectrum beta-lactamase phenotype (ESBLP) and with resistance to aminoglycosides, chloramphenicol, fluoroquinolones, nitrofurantoin, tiamulin, and trimethoprim/sulfamethoxazole were significantly more common among the PA farms (p < 0.05) than in the other two farm categories. In the PA farms, ESBLP E. coli commonly contained the blaCTX-M-1 group, blaCTX-M-9 group, or both gene groups, and were shown to transfer blaCTX-M genes in a conjugation experiment. E. coli containing N, FIC and A/C replicons were found only in PA farms. In summary, although E. coli isolates from all farms contained a core set of resistance to β-lactams and tetracyclines, the routine use of PA increased resistance rates to other important antimicrobials.
Get full access to this article
View all access options for this article.
References
1.
European Food Safety Authority (EFSA). 2012. The European Union Summary Report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food. EFSA J., 10:2598.
2.
World Health Organization (WHO). 2014. Antimicrobial Resistance Global Report on Surveillance. WHO Annual Report, Avenue Appia, Geneva, Switzerland.
3.
LayK.K., KoowattananukulC., ChansongN., and ChuanchuenR.. 2012. Antimicrobial resistance, virulence, and phylogenetic characteristics of Escherichia coli isolates from clinically healthy swine. Foodborne Pathog. Dis., 9:992–1001.
4.
PrapasarakulN., TummarukP., NiyomtumW., TripipatT., and SerichantalergsO.. 2010. Virulence genes and antimicrobial susceptibilities of hemolytic and nonhemolytic Escherichia coli isolated from post-weaning piglets in central Thailand. J. Vet. Med. Sci., 72:1603–1608.
5.
DohmenW., BontenM.J., BosM.E., van MarmS., ScharringaJ., WagenaarJ.A., and HeederikD.J.. 2015. Carriage of extended-spectrum beta-lactamases in pig farmers is associated with occurrence in pigs. Clin. Microbiol. Infect., 21:917–923.
6.
AgersoY., AarestrupF.M., PedersenK., SeyfarthA.M., StruveT., and HasmanH.. 2012. Prevalence of extended-spectrum cephalosporinase (ESC)-producing Escherichia coli in Danish slaughter pigs and retail meat identified by selective enrichment and association with cephalosporin usage. J. Antimicrob. Chemother., 67:582–588.
7.
CavacoL.M., AbatihE., AarestrupF.M., and GuardabassiL.. 2008. Selection and persistence of CTX-M-producing Escherichia coli in the intestinal flora of pigs treated with amoxicillin, ceftiofur, or cefquinome. Antimicrob. Agents Chemother., 52:3612–3616.
8.
ZhouL.J., YingG.G., ZhangR.Q., LiuS., LaiH.J., ChenZ.F., YangB., and ZhaoJ.L.. 2013. Use patterns, excretion masses and contamination profiles of antibiotics in a typical swine farm, south China. Environ. Sci. Process Impacts., 15:802–813.
9.
MakitaK., GotoM., OzawaM., KawanishiM., KoikeR., AsaiT., and TamuraY.. 2016. Multivariable analysis of the association between antimicrobial use and antimicrobial resistance in Escherichia coli isolated from apparently healthy pigs in Japan. Microb. Drug Resist., 22:28–39.
10.
RenouxG., and TerdjmanA.. 1951. Clinical value of the precise identification of gram-negative urinary bacilli; practical value of the medium eosine-methylene blue (E.M.B.)]. Presse Med., 59:203–206.
11.
ClarkH.F., GeldreichE.E., KablerP.W., BordnerR.H., and HuffC.B.. 1957. The coliform group. I. The boric acid lactose broth reaction of coliform IMViC types. Appl. Microbiol., 5:396–400.
12.
ClermontO., ChristensonJ.K., DenamurE., and GordonD.M.. 2013. Ther Clermont Escherichia coli phylo-typing method revisited: improvement of specificity and detection of new phylo-groups. Environ. Microb. Rep., 5:58–65.
13.
Plumb. 2015. Plumb's Veterinary Drug Handbook. 8th ed. Wiley-Blackwell Press, Indianapolis, IN.
14.
Copyright Division of Animal Feed and Veterinary Products Control 1999. Act of legislation: drugs for animals usage, The Department of Livestock Development (DLD), Ministry of Agriculture and Cooperatives (MoAC), Bangkok, Thailand.
15.
Clinical and Laboratory Standards Institute (CLSI). 2015. Performance Standards for Antimicrobial Disk and Dilution Susceptibility test for Bacteria Isolated From Animals; 3rd VET01-S3. Clinical and Laboratory Standards Institute, Wayne, PA.
16.
Clinical and Laboratory Standards Institute (CLSI). 2015. Performance Standards for Antimicrobial Susceptibility Testing; 24th Informational Supplement. M100-S25. Clinical and Laboratory Standards Institute, Wayne, PA.
17.
EspinarM.J., RochaR., RibeiroM., Goncalves RodriguesA., and Pina-VazC.. 2011. Extended-spectrum beta-lactamases of Escherichia coli and Klebsiella pneumoniae screened by the VITEK 2 system. J. Med. Microbiol., 60:756–760.
18.
XuL., EnsorV., GossainS., NyeK., and HawkeyP.. 2005. Rapid and simple detection of blaCTX-M genes by multiplex PCR assay. J. Med. Microbiol., 54:1183–1187.
19.
GrayK.J., WilsonL.K., PhiriA., CorkillJ.E., FrenchN., and HartC.A.. 2006. Identification and characterization of ceftriaxone resistance and extended-spectrum beta-lactamases in Malawian bacteraemic Enterobacteriaceae. J. Antimicrob. Chemother., 57:661–665.
20.
ChuanchuenR., AjariyakhajornK., KoowatananukulC., WannaprasatW., KhemtongS., and SamngamnimS.. 2010. Antimicrobial resistance and virulence genes in Salmonella enterica isolates from dairy cows. Foodborne Pathog. Dis., 7:63–69.
21.
CarattoliA., BertiniA., VillaL., FalboV., HopkinsK.L., and ThrelfallE.J.. 2005. Identification of plasmids by PCR-based replicon typing. J. Microbiol Methods., 63:219–228.
22.
GoldsteinC., LeeM.D., SanchezS., HudsonC., PhillipsB., RegisterB., GradyM., LiebertC., SummersA.O., WhiteD.G., et al.2001. Incidence of class 1 and 2 integrases in clinical and commensal bacteria from livestock, companion animals, and exotics. Antimicrob. Agents Chemother., 45:723–726.
23.
ChangkaewK., IntarapukA., UtrarachkijF., NakajimaC., SuthienkulO., and SuzukiY.. 2015. Antimicrobial resistance, extended-spectrum beta-lactamase productivity, and class 1 integrons in Escherichia coli from healthy swine. J. Food Prot., 78:1442–1450.
24.
KhamsarnS., NampoonsakY., BusamaroS., TangkoskulT., SeenamaC., RattanaumpawanP., BoonyasiriA., and ThamlikitkulV.. 2016. Epidemiology of antibiotic use and antimicrobial resistance in selected communities in Thailand. J. Med. Assoc. Thai., 99:270–275.
25.
NhungN.T., CuongN.V., ThwaitesG., and Carrique-MasJ.. 2016. Antimicrobial usage in Asia: a review. Antibiotics (Basel)., 5:1–24.
26.
TrongjitS., AngkittitrakulS., and ChuanchuenR.. 2016. Occurrence and molecular characteristics of antimicrobial resistance of Escherichia coli from broilers, pigs and meat products in Thailand and Cambodia provinces. Microbiol. Immunol., 60:575–585.
27.
EnneV.I., DelsolA.A., DavisG.R., HaywardS.L., RoeJ.M., and BennettP.M.. 2005. Assessment of the fitness impacts on Escherichia coli of acquisition of antibiotic resistance genes encoded by different types of genetic element. J. Antimicrob. Chemother., 56:544–551.
28.
KirchnerM., MafuraM., HuntT., Abu-OunM., Nunez-GarciaJ., HuY., WeileJ., CoatesA., CardR., and AnjumM.F.. 2014. Antimicrobial resistance characteristics and fitness of Gram-negative fecal bacteria from volunteers treated with minocycline or amoxicillin. Front Microbiol., 5:1–9.
29.
Ministry of Agriculture Cooperatives of Thailand. 2009. Procedure of animal drug usage control Ministry Annoucement. Ministry of Agriculture and Cooperatives (MoAC), Bangkok, Thailand.
30.
JohnsonT.A., StedtfeldR.D., WangQ., ColeJ.R., HashshamS.A., LooftT., ZhuY.G., and TiedjeJ.M.. 2016. Clusters of antibiotic resistance genes enriched together stay together in swine agriculture. MBio., 7:e02214–e02215.
31.
van DuijkerenE., GrekoC., PringleM., BaptisteK.E., CatryB., JukesH., MorenoM.A., PombaM.C., PyoralaS., RantalaM., et al.2014. Pleuromutilins: use in food-producing animals in the European Union, development of resistance and impact on human and animal health. J. Antimicrob. Chemother., 69:2022–2031.
32.
LongK.S., PoehlsgaardJ., KehrenbergC., SchwarzS., and VesterB.. 2006. The cfr rRNA methyltransferase confers resistance to Phenicols, Lincosamides, Oxazolidinones, Pleuromutilins, and Streptogramin A antibiotics. Antimicrob. Agents Chemother., 50:2500–2505.
33.
KadlecK., and SchwarzS.. 2008. Analysis and distribution of class 1 and class 2 integrons and associated gene cassettes among Escherichia coli isolates from swine, horses, cats and dogs collected in the BfT-GermVet monitoring study. J. Antimicrob. Chemother., 62:469–473.
34.
PovilonisJ., SeputieneV., RuzauskasM., SiugzdinieneR., VirgailisM., PavilonisA., and SuziedelieneE.. 2010. Transferable class 1 and 2 integrons in Escherichia coli and Salmonella enterica isolates of human and animal origin in Lithuania. Foodborne Pathog. Dis., 7:1185–1192.
35.
SzmolkaA., and NagyB.. 2013. Multidrug resistant commensal Escherichia coli in animals and its impact for public health. Front Microbiol., 4:1–13.
36.
SchmithausenR.M., Schulze-GeisthoevelS.V., StemmerF., El-JadeM., ReifM., HackS., MeilaenderA., MontabauerG., FimmersR., ParcinaM., et al.2015. Analysis of transmission of MRSA and ESBL-E among pigs and farm personnel. PLoS One., 10:1–24.
37.
ZhengH., ZengZ., ChenS., LiuY., YaoQ., DengY., ChenX., LvL., ZhuoC., ChenZ., et al.2012. Prevalence and characterisation of CTX-M beta-lactamases amongst Escherichia coli isolates from healthy food animals in China. Int. J. Antimicrob. Agents., 39:305–310.
38.
BoonyasiriA., TangkoskulT., SeenamaC., SaiyarinJ., TiengrimS., and ThamlikitkulV.. 2014. Prevalence of antibiotic resistant bacteria in healthy adults, foods, food animals, and the environment in selected areas in Thailand. Pathog. Glob. Health., 108:235–245.
39.
KiratisinP., ApisarnthanarakA., LaesripaC., and SaifonP.. 2008. Molecular characterization and epidemiology of extended-spectrum-beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolates causing health care-associated infection in Thailand, where the CTX-M family is endemic. Antimicrob. Agents Chemother., 52:2818–2824.
40.
KoY.J., MoonH.W., HurM., ParkC.M., ChoS.E., and YunY.M.. 2013. Fecal carriage of extended-spectrum beta-lactamase-producing Enterobacteriaceae in Korean community and hospital settings. Infection., 41:9–13.
41.
NakaneK., KawamuraK., GotoK., and ArakawaY.. 2016. Long-term colonization by bla(CTX-M)-harboring Escherichia coli in healthy Japanese people engaged in food handling. Appl. Environ. Microbiol., 82:1818–1827.
42.
MoodleyA., and GuardabassiL.. 2009. Transmission of IncN plasmids carrying blaCTX-M-1 between commensal Escherichia coli in pigs and farm workers. Antimicrob. Agents Chemother., 53:1709–1711.
43.
ZurfluhK., JakobiG., StephanR., HachlerH., and Nuesch-InderbinenM.. 2014. Replicon typing of plasmids carrying blaCTX-M-1 in Enterobacteriaceae of animal, environmental and human origin. Front Microbiol., 5:1–5.
44.
JohnsonT.J., WannemuehlerY.M., JohnsonS.J., LogueC.M., WhiteD.G., DoetkottC., and NolanL.K.. 2007. Plasmid replicon typing of commensal and pathogenic Escherichia coli isolates. Appl. Environ. Microbiol., 73:1976–1983.
45.
MshanaS.E., ImirzaliogluC., HossainH., HainT., DomannE., and ChakrabortyT.. 2009. Conjugative IncFI plasmids carrying CTX-M-15 among Escherichia coli ESBL producing isolates at a University hospital in Germany. BMC Infect. Dis., 9:1–8.
46.
CarattoliA.2009. Resistance plasmid families in Enterobacteriaceae. Antimicrob. Agents Chemother., 53:2227–2238.
47.
CottellJ.L., KanwarN., Castillo-CourtadeL., ChalmersG., ScottH.M., NorbyB., LoneraganG.H., and BoerlinP.. 2013. blaCTX-M-32 on an IncN plasmid in Escherichia coli from beef cattle in the United States. Antimicrob. Agents Chemother., 57:1096–1097.
48.
JakobsenL., BortolaiaV., BielakE., MoodleyA., OlsenS.S., HansenD.S., Frimodt-MollerN., GuardabassiL., and HasmanH.. 2015. Limited similarity between plasmids encoding CTX-M-1 beta-lactamase in Escherichia coli from humans, pigs, cattle, organic poultry layers and horses in Denmark. J. Glob. Antimicrob. Resist., 3:132–136.
49.
KayamaS., ShigemotoN., KuwaharaR., OshimaK., HirakawaH., HisatsuneJ., JoveT., NishioH., YamasakiK., WadaY., et al.2015. Complete nucleotide sequence of the IncN plasmid encoding IMP-6 and CTX-M-2 from emerging carbapenem-resistant Enterobacteriaceae in Japan. Antimicrob. Agents Chemother., 59:1356–1359.
50.
HumphreyB., ThomsonN.R., ThomasC.M., BrooksK., SandersM., DelsolA.A., RoeJ.M., BennettP.M., and EnneV.I.. 2012. Fitness of Escherichia coli strains carrying expressed and partially silent IncN and IncP1 plasmids. BMC Microbiol., 12:1–9.
51.
HuangS.Y., ZhuX.Q., WangY., LiuH.B., DaiL., HeJ.K., LiB.B., WuC.M., and ShenJ.Z.. 2012. Co-carriage of qnrS1, floR, and blaCTX-M-14 on a multidrug-resistant plasmid in Escherichia coli isolated from pigs. Foodborne Pathog. Dis., 9:896–901.
52.
LeeK.E., LimS.I., ChoiH.W., LimS.K., SongJ.Y., and AnD.J.. 2014. Plasmid-mediated AmpC beta-lactamase (CMY-2) gene in Salmonella Typhimurium isolated from diarrheic pigs in South Korea. BMC Res. Notes., 7:1–4.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
