Background Plasmid mediated antimicrobial resistance in the Enterobacteriaceae is a global problem. The rise of CTX-M class extended spectrum beta lactamases (ESBLs) has been well documented in industrialized countries. Vietnam is representative of a typical transitional middle income country where the spectrum of infectious diseases combined with the spread of drug resistance is shifting and bringing new healthcare challenges. Methodology We collected hospital admission data from the pediatric population attending the hospital for tropical diseases in Ho Chi Minh City with Shigella infections. Organisms were cultured from all enrolled patients and subjected to antimicrobial susceptibility testing. Those that were ESBL positive were subjected to further investigation. These investigations included PCR amplification for common ESBL genes, plasmid investigation, conjugation, microarray hybridization and DNA sequencing of a blaCTX–M encoding plasmid. Principal Findings We show that two different blaCTX-M genes are circulating in this bacterial population in this location. Sequence of one of the ESBL plasmids shows that rather than the gene being integrated into a preexisting MDR plasmid, the blaCTX-M gene is located on relatively simple conjugative plasmid. The sequenced plasmid (pEG356) carried the blaCTX-M-24 gene on an ISEcp1 element and demonstrated considerable sequence homology with other IncFI plasmids. Significance The rapid dissemination, spread of antimicrobial resistance and changing population of Shigella spp. concurrent with economic growth are pertinent to many other countries undergoing similar development. Third generation cephalosporins are commonly used empiric antibiotics in Ho Chi Minh City. We recommend that these agents should not be considered for therapy of dysentery in this setting.
References
[1]
Livermore DM, Canton R, Gniadkowski M, Nordmann P, Rossolini GM, et al. (2007) CTX-M: changing the face of ESBLs in Europe. J Antimicrob Chemother 59: 165–174. doi: 10.1093/jac/dkl483
[2]
Rossolini GM, D'Andrea MM, Mugnaioli C (2008) The spread of CTX-M-type extended-spectrum beta-lactamases. Clin Microbiol Infect 14: Suppl 133–41. doi: 10.1111/j.1469-0691.2007.01867.x
[3]
Heffernan HM, Woodhouse RE, Pope CE, Blackmore TK (2009) Prevalence and types of extended-spectrum beta-lactamases among urinary Escherichia coli and Klebsiella spp. in New Zealand. Int J Antimicrob Agents. doi: 10.1016/j.ijantimicag.2009.07.014
Valverde A, Coque TM, Garcia-San Miguel L, Baquero F, Canton R (2008) Complex molecular epidemiology of extended-spectrum beta-lactamases in Klebsiella pneumoniae: a long-term perspective from a single institution in Madrid. J Antimicrob Chemother 61: 64–72. doi: 10.1093/jac/dkm403
[6]
Kiratisin P, Apisarnthanarak A, Laesripa C, Saifon P (2008) Molecular characterization and epidemiology of extended-spectrum-beta-lactamase-produci？ngEscherichia 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. doi: 10.1128/AAC.00171-08
[7]
Ruppe E, Hem S, Lath S, Gautier V, Ariey F, et al. (2009) CTX-M beta-lactamases in Escherichia coli from community-acquired urinary tract infections, Cambodia. Emerg Infect Dis 15: 741–748. doi: 10.3201/eid1505.071299
[8]
Le TM, Baker S, Le TP, Le TP, Cao TT, et al. (2009) High prevalence of plasmid-mediated quinolone resistance determinants in commensal members of the Enterobacteriaceae in Ho Chi Minh City, Vietnam. J Med Microbiol 58: 1585–1592. doi: 10.1099/jmm.0.010033-0
[9]
Kotloff KL, Winickoff JP, Ivanoff B, Clemens JD, Swerdlow DL, et al. (1999) Global burden of Shigella infections: implications for vaccine development and implementation of control strategies. Bull World Health Organ 77: 651–666.
[10]
Kim DR, Ali M, Thiem VD, Park JK, von Seidlein L, et al. (2008) Geographic analysis of shigellosis in Vietnam. Health Place 14: 755–767. doi: 10.1016/j.healthplace.2007.12.003
[11]
Vinh H, Wain J, Chinh MT, Tam CT, Trang PT, et al. (2000) Treatment of bacillary dysentery in Vietnamese children: two doses of ofloxacin versus 5-days nalidixic acid. Trans R Soc Trop Med Hyg 94: 323–326. doi: 10.1016/S0035-9203(00)90343-2
[12]
WHO (2005) Shigellosis: disease burden, epidemiology and case management. Weekly epidemiological record 93–100.
[13]
Chau TT, Campbell JI, Galindo CM, Van Minh Hoang N, Diep TS, et al. (2007) Antimicrobial drug resistance of Salmonella enterica serovar typhi in asia and molecular mechanism of reduced susceptibility to the fluoroquinolones. Antimicrob Agents Chemother 51: 4315–4323. doi: 10.1128/AAC.00294-07
[14]
Hu LF, Li JB, Ye Y, Li X (2007) Mutations in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV in clinical strains of fluoroquinolone-resistant Shigella in Anhui, China. J Microbiol 45: 168–170.
[15]
Vinh H, Nhu NT, Nga TV, Duy PT, Campbell JI, et al. (2009) A changing picture of shigellosis in southern Vietnam: shifting species dominance, antimicrobial susceptibility and clinical presentation. BMC Infect Dis 9: 204. doi: 10.1186/1471-2334-9-204
[16]
Iversen J, Sandvang D, Srijan A, Cam PD, Dalsgaard A (2003) Characterization of antimicrobial resistance, plasmids, and gene cassettes in Shigella spp. from patients in vietnam. Microb Drug Resist 9: Suppl 1S17–24. doi: 10.1089/107662903322541856
[17]
Bratoeva MP, John JF Jr (1994) In vivo R-plasmid transfer in a patient with a mixed infection of shigella dysentery. Epidemiol Infect 112: 247–252. doi: 10.1017/S0950268800057654
[18]
Dutta S, Rajendran K, Roy S, Chatterjee A, Dutta P, et al. (2002) Shifting serotypes, plasmid profile analysis and antimicrobial resistance pattern of shigellae strains isolated from Kolkata, India during 1995–2000. Epidemiol Infect 129: 235–243. doi: 10.1017/S0950268802007240
[19]
von Seidlein L, Kim DR, Ali M, Lee H, Wang X, et al. (2006) A multicentre study of Shigella diarrhoea in six Asian countries: disease burden, clinical manifestations, and microbiology. PLoS Med 3: e353. doi: 10.1371/journal.pmed.0030353
[20]
Kuo CY, Su LH, Perera J, Carlos C, Tan BH, et al. (2008) Antimicrobial susceptibility of Shigella isolates in eight Asian countries, 2001–2004. J Microbiol Immunol Infect 41: 107–111.
[21]
Anh NT, Cam PD, Dalsgaard A (2001) Antimicrobial resistance of Shigella spp isolated from diarrheal patients between 1989 and 1998 in Vietnam. Southeast Asian J Trop Med Public Health 32: 856–862.
[22]
Lartigue MF, Poirel L, Decousser JW, Nordmann P (2005) Multidrug-resistant Shigella sonnei and Salmonella enterica Serotype typhimurium isolates producing CTX-M beta-lactamases as causes of community-acquired infection in France. Clin Infect Dis 40: 1069–1070. doi: 10.1086/428667
[23]
Jarlier V, Nicolas MH, Fournier G, Philippon A (1988) Extended broad-spectrum beta-lactamases conferring transferable resistance to newer beta-lactam agents in Enterobacteriaceae: hospital prevalence and susceptibility patterns. Rev Infect Dis 10: 867–878. doi: 10.1093/clinids/10.4.867
[24]
Thomson KS, Sanders CC (1992) Detection of extended-spectrum beta-lactamases in members of the family Enterobacteriaceae: comparison of the double-disk and three-dimensional tests. Antimicrob Agents Chemother 36: 1877–1882. doi: 10.1128/AAC.36.9.1877
[25]
al Naiemi N, Duim B, Bart A (2006) A CTX-M extended-spectrum beta-lactamase in Pseudomonas aeruginosa and Stenotrophomonas maltophilia. J Med Microbiol 55: 1607–1608. doi: 10.1099/jmm.0.46704-0
Baker S, Campbell JI, Stabler R, Nguyen HV, To DS, et al. (2008) Fatal wound infection caused by Chromobacterium violaceum in Ho Chi Minh City, Vietnam. J Clin Microbiol 46: 3853–3855. doi: 10.1128/JCM.01068-08
[28]
Stabler RA, Dawson LF, Oyston PC, Titball RW, Wade J, et al. (2008) Development and application of the active surveillance of pathogens microarray to monitor bacterial gene flux. BMC Microbiol 8: 177. doi: 10.1186/1471-2180-8-177
[29]
Hinchliffe SJ, Isherwood KE, Stabler RA, Prentice MB, Rakin A, et al. (2003) Application of DNA microarrays to study the evolutionary genomics of Yersinia pestis and Yersinia pseudotuberculosis. Genome Res 13: 2018–2029. doi: 10.1101/gr.1507303
[30]
Kado CI, Liu ST (1981) Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145: 1365–1373.
[31]
Batchelor M, Hopkins K, Threlfall EJ, Clifton-Hadley FA, Stallwood AD, et al. (2005) bla(CTX-M) genes in clinical Salmonella isolates recovered from humans in England and Wales from 1992 to 2003. Antimicrob Agents Chemother 49: 1319–1322. doi: 10.1128/AAC.49.4.1319-1322.2005
[32]
Xiong Z, Li T, Xu Y, Li J (2007) Detection of CTX-M-14 extended-spectrum beta-lactamase in Shigella sonnei isolates from China. J Infect 55: e125–128. doi: 10.1016/j.jinf.2007.07.017
[33]
Parkhill J, Achtman M, James KD, Bentley SD, Churcher C, et al. (2000) Complete DNA sequence of a serogroup A strain of Neisseria meningitidis Z2491. Nature 404: 502–506. doi: 10.1038/35006655
[34]
Fricke WF, McDermott PF, Mammel MK, Zhao S, Johnson TJ, et al. (2009) Antimicrobial resistance-conferring plasmids with similarity to virulence plasmids from avian pathogenic Escherichia coli strains in Salmonella enterica serovar Kentucky isolates from poultry. Appl Environ Microbiol 75: 5963–5971. doi: 10.1128/AEM.00786-09
[35]
Carver TJ, Rutherford KM, Berriman M, Rajandream MA, Barrell BG, et al. (2005) ACT: the Artemis Comparison Tool. Bioinformatics 21: 3422–3423. doi: 10.1093/bioinformatics/bti553
[36]
Carver T, Thomson N, Bleasby A, Berriman M, Parkhill J (2009) DNAPlotter: circular and linear interactive genome visualization. Bioinformatics 25: 119–120. doi: 10.1093/bioinformatics/btn578
[37]
Parry CM, Duong NM, Zhou J, Mai NT, Diep TS, et al. (2002) Emergence in Vietnam of Streptococcus pneumoniae resistant to multiple antimicrobial agents as a result of dissemination of the multiresistant Spain(23F)-1 clone. Antimicrob Agents Chemother 46: 3512–3517. doi: 10.1128/AAC.46.11.3512-3517.2002
[38]
Vinh H, Baker S, Campbell J, Hoang NV, Loan HT, et al. (2009) Rapid emergence of third generation cephalosporin resistant Shigella spp. in Southern Vietnam. J Med Microbiol 58: 281–283. doi: 10.1099/jmm.0.002949-0
[39]
Nguyen TV, Le PV, Le CH, Weintraub A (2005) Antibiotic resistance in diarrheagenic Escherichia coli and Shigella strains isolated from children in Hanoi, Vietnam. Antimicrob Agents Chemother 49: 816–819. doi: 10.1128/AAC.49.2.816-819.2005
[40]
Bonnet R (2004) Growing group of extended-spectrum beta-lactamases: the CTX-M enzymes. Antimicrob Agents Chemother 48: 1–14. doi: 10.1128/AAC.48.1.1-14.2004
[41]
Lee SG, Jeong SH, Lee H, Kim CK, Lee Y, et al. (2009) Spread of CTX-M-type extended-spectrum beta-lactamases among bloodstream isolates of Escherichia coli and Klebsiella pneumoniae from a Korean hospital. Diagn Microbiol Infect Dis 63: 76–80. doi: 10.1016/j.diagmicrobio.2008.09.002
[42]
Yuan L, Liu JH, Hu GZ, Pan YS, Liu ZM, et al. (2009) Molecular characterization of extended-spectrum beta-lactamase-producing Escherichia coli isolates from chickens in Henan Province, China. J Med Microbiol 58: 1449–1453. doi: 10.1099/jmm.0.012229-0
[43]
Pitout JD, Gregson DB, Campbell L, Laupland KB (2009) Molecular characteristics of extended-spectrum-beta-lactamase-produci？ngEscherichia coli isolates causing bacteremia in the Calgary Health Region from 2000 to 2007: emergence of clone ST131 as a cause of community-acquired infections. Antimicrob Agents Chemother 53: 2846–2851. doi: 10.1128/AAC.00247-09
[44]
Shen P, Jiang Y, Zhou Z, Zhang J, Yu Y, et al. (2008) Complete nucleotide sequence of pKP96, a 67 850 bp multiresistance plasmid encoding qnrA1, aac(6′)-Ib-cr and blaCTX-M-24 from Klebsiella pneumoniae. J Antimicrob Chemother 62: 1252–1256. doi: 10.1093/jac/dkn397
[45]
Saadi S, Maas WK, Bergquist PL (1984) RepFIC, a basic replicon of IncFI plasmids that has homology with a basic replicon of IncFII plasmids. Plasmid 12: 61–64. doi: 10.1016/0147-619X(84)90067-2
[46]
Woodford N, Carattoli A, Karisik E, Underwood A, Ellington MJ, et al. (2009) Complete nucleotide sequences of plasmids pEK204, pEK499, and pEK516, encoding CTX-M enzymes in three major Escherichia coli lineages from the United Kingdom, all belonging to the international O25:H4-ST131 clone. Antimicrob Agents Chemother 53: 4472–4482. doi: 10.1128/AAC.00688-09
[47]
Chen YT, Lauderdale TL, Liao TL, Shiau YR, Shu HY, et al. (2007) Sequencing and comparative genomic analysis of pK29, a 269-kilobase conjugative plasmid encoding CMY-8 and CTX-M-3 beta-lactamases in Klebsiella pneumoniae. Antimicrob Agents Chemother 51: 3004–3007. doi: 10.1128/AAC.00167-07
[48]
Chanawong A, M'Zali FH, Heritage J, Xiong JH, Hawkey PM (2002) Three cefotaximases, CTX-M-9, CTX-M-13, and CTX-M-14, among Enterobacteriaceae in the People's Republic of China. Antimicrob Agents Chemother 46: 630–637. doi: 10.1128/AAC.46.3.630-637.2002
[49]
Cao V, Lambert T, Nhu DQ, Loan HK, Hoang NK, et al. (2002) Distribution of extended-spectrum beta-lactamases in clinical isolates of Enterobacteriaceae in Vietnam. Antimicrob Agents Chemother 46: 3739–3743. doi: 10.1128/AAC.46.12.3739-3743.2002
[50]
Koh TH, Wang GC, Sng LH, Koh TY (2004) CTX-M and plasmid-mediated AmpC-producing Enterobacteriaceae Singapore. Emerg Infect Dis 10: 1172–1174. doi: 10.3201/eid1006.030726
[51]
Radice M, Gonzealez C, Power P, Vidal MC, Gutkind G (2001) Third-generation cephalosporin resistance in Shigella sonnei, Argentina. Emerg Infect Dis 7: 442–443. doi: 10.3201/eid0703.010313
[52]
Kim S, Kim J, Kang Y, Park Y, Lee B (2004) Occurrence of extended-spectrum beta-lactamases in members of the genus Shigella in the Republic of Korea. J Clin Microbiol 42: 5264–5269. doi: 10.1128/JCM.42.11.5264-5269.2004
[53]
Hrabak J, Empel J, Gniadkowski M, Halbhuber Z, Rebl K, et al. (2008) CTX-M-15-producing Shigella sonnei strain from a Czech patient who traveled in Asia. J Clin Microbiol 46: 2147–2148. doi: 10.1128/JCM.00427-08
[54]
Nagano Y, Nagano N, Wachino J, Ishikawa K, Arakawa Y (2009) Novel chimeric beta-lactamase CTX-M-64, a hybrid of CTX-M-15-like and CTX-M-14 beta-lactamases, found in a Shigella sonnei strain resistant to various oxyimino-cephalosporins, including ceftazidime. Antimicrob Agents Chemother 53: 69–74. doi: 10.1128/AAC.00227-08
[55]
Pitout JD, Campbell L, Church DL, Gregson DB, Laupland KB (2009) Molecular characteristics of travel-related extended-spectrum-beta-lactamase-produci？ngEscherichia coli isolates from the Calgary Health Region. Antimicrob Agents Chemother 53: 2539–2543. doi: 10.1128/AAC.00061-09
[56]
Wirth T, Falush D, Lan R, Colles F, Mensa P, et al. (2006) Sex and virulence in Escherichia coli: an evolutionary perspective. Mol Microbiol 60: 1136–1151. doi: 10.1111/j.1365-2958.2006.05172.x
[57]
Holt KE, Parkhill J, Mazzoni CJ, Roumagnac P, Weill FX, et al. (2008) High-throughput sequencing provides insights into genome variation and evolution in Salmonella typhi. Nat Genet 40: 987–993. doi: 10.1038/ng.195
