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Six imported pigs originating from Guangdong, Henan, and Hunan provinces in China during October 2015 to February 2017 were cultured and found to be positive for meropenem-resistant Escherichia coli The samples yielded 9 E. coli isolates of diverse sequence types carrying bla NDM-5 on IncX3 (8 isolates from 5 farms) or IncFII (1 isolate from 1 farm) plasmids. The mcr-1 gene was coharbored by 4 isolates. The IncX3 plasmids (∼46 kb) carrying bla NDM-5 were identical or nearly identical to each other., (Copyright © 2018 American Society for Microbiology.)

This study investigated 248 extended-spectrum β-lactamase-producing Escherichia coli isolates from 2012 to 2013 for hybrid blaCTX-M genes. blaCTX-M genes were detected in 228 isolates of which 14 isolates were hybrid blaCTX-M positive (6 blaCTX-M-123, 6 blaCTX-M-64, and 2 blaCTX-M-132). The 14 hybrid blaCTX-M-carrying isolates (8 from chickens, 2 each from pigs and cattle, 1 each from dog and rodent) were genetically diverse. All but 2 hybrid blaCTX-M were carried on IncI1 (5 blaCTX-M-123) and IncI2 (6 blaCTX-M-64 and one blaCTX-M-132) plasmids. Our IncI1 and IncI2 plasmids had pHNAH4-1-like and pHN1122-1-like restriction fragment length polymorphism patterns, respectively. Genetic relatedness of the plasmids to pHNAH4-1 and pHN1122-1 were confirmed by complete sequencing of 3 plasmids, pCTXM123_C0996, pCTXM64_C0967, and pCTXM132_P0421. Plasmids closely related to pHNAH4-1 and pHN1122-1 and carrying different blaCTX-M alleles have been reported from multiple geographic areas in China previously. The findings highlighted the wide dissemination of hybrid blaCTX-M variants in different parts of China., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Previous work on the subclones within Escherichia coli ST131 predominantly involved isolates from Western countries. This study assessed the prevalence and antimicrobial resistance attributed to this clonal group. A total of 340 consecutive, non-duplicated urinary E. coli isolates originating from four clinical laboratories in Hong Kong in 2013 were tested. ST131 prevalence among the total isolates was 18.5 % (63/340) and was higher among inpatient isolates (23.0 %) than outpatient isolates (11.8 %, P<0.001), and higher among isolates from patients aged ≥65 years than from patients aged 18-50 years and 51-64 years (25.4 vs 3.4 and 4.0 %, respectively, P<0.001). Of the 63 ST131 isolates, 43 (68.3 %) isolates belonged to the H30 subclone, whereas the remaining isolates belonged to H41 (n = 17), H54 (n = 2) and H22 (n = 1). All H30 isolates were ciprofloxacin-resistant, of which 18.6 % (8/43) belonged to the H30-Rx subclone. Twenty-six (41.3 %) ST131 isolates were ESBL-producers, of which 19 had blaCTX-M-14 (12 non-H30-Rx, two H30-Rx and five H41), six had blaCTX-M-15 (five non-H30-Rx and one H30-Rx) and one was blaCTX-M-negative (H30). In conclusion, ST131 accounts for a large share of the antimicrobial-resistant E. coli isolates from geriatric patients. Unlike previous reports, ESBL-producing ST131 strains mainly belonged to non-H30-Rx rather than the H30-Rx subclone, with blaCTX-M-14 as the dominant enzyme type., (© 2015 The Authors.)

This territory-wide study investigated the occurrence of faecal carriage of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli among wild rodents from the 18 districts in Hong Kong. Individual rectal swabs were obtained from the trapped animals and cultured in plain and selective media. A total of 965 wild rodents [148 chestnut spiny rats (Niviventer fulvescens), 326 Indo-Chinese forest rats (Rattus andamanensis), 452 brown rats (Rattus norvegicus) and 39 black rats (Rattus rattus)] were sampled. ESBL carriage was 0 % in chestnut spiny rats, 0.6 % in Indo-Chinese forest rats, 7.7 % in black rats and 13.9 % in brown rats. Among brown rats, the prevalence of ESBL carriage differed markedly by geographical location: absent in two districts, low (7-10 %) in six districts, moderate (11-19 %) in seven districts and high (21-50 %) in three districts. Nonetheless, there was no correlation between the prevalence of ESBL in brown rats and human population density in the 18 districts. CTX-M-type enzymes were detected in 92.0 % of the ESBL-producing isolates, of which 83.1 % were resistant to three or more non-β-lactam drugs. The CTX-M producing isolates were genetically diverse but a large proportion (47.8 %) were included in six successful clones that are strongly associated with human diseases and CTX-M dissemination, viz. sequence type complex [STC]10/phylogroup A, STC23/phylogroup B1, STC38/phylogroup D, STC155/phylogroup B1, ST405/phylogroup D and ST131/phylogroup B2. In conclusion, our results show that brown rats often carry potentially zoonotic clones of CTX-M producing, multidrug-resistant E. coli. The potential for rats to be a source of CTX-M producing E. coli for humans deserves further consideration., (© 2015 The Authors.)

Few studies have described how an expanding elderly population influences the burden of antimicrobial resistance in micro-organisms. This study aimed to investigate trends in age-stratified extended-spectrum β-lactamase (ESBL)-positive Escherichia coli metrics in relation to an ageing population. The antimicrobial resistance database of E. coli from a healthcare region in Hong Kong from 2003 to 2012 was retrospectively reviewed. Future trends in age-stratified ESBL metrics were predicted up to 2022. Susceptibility results of clinical E. coli isolates from patients aged 0-74 years (n = 17 853) and aged ≥75 years (n = 17 047) were analysed. For the period 2003-2012, 23.7 % of the hospital admissions were of patients aged ≥75 years. However, approximately half of the annual ESBL-positive E. coli isolates were recovered from patients aged ≥75 years, being 55.0 % (233/424) in 2003 and 56.0 % (639/1142) in 2012. During this period of time, the annual prevalence and cumulative incidence of ESBL-positive E. coli in patients aged ≥75 years were significantly higher than in patients aged 0-74 years. From 2012-2022, it is predicted that ESBL-positive E. coli prevalence among patients aged 0-74 years and ≥75 years would increase from 25.4 % to 50.2 % and from 30.8 % to 70.0 %, respectively. In 2022, the predicted ESBL-positive E. coli cumulative incidence would be 63.7 per 10 000 admissions and 178.7 per 10 000 admissions among patients aged 0-74 years and ≥75 years, respectively. In conclusion, a rapidly expanding elderly population would substantially add to the burden of ESBL., (© 2014 The Authors.)

A total of 1878 non-duplicate clinical Escherichia coli isolates (comprising 1711 urinary isolates and 167 blood-culture isolates), which were collected from multiple centres in Hong Kong during 1996-2008, were used to investigate the prevalence and molecular epidemiology of plasmid-mediated fosfomycin (fos) resistance genes. Eighteen of the 1878 clinical E. coli isolates were fosfomycin resistant, of which six were fosA3 positive and two were positive for another fosA variant (designated fosKP96). No isolates had the fosC2 gene. The clones of the eight isolates were diverse: sequence type (ST) 95 (n = 2), ST118 (n = 1), ST131 (n = 1), ST617 (n = 1), ST648 (n = 1), ST1488 (n = 1) and ST2847 (n = 1). In the isolates, fosA3 and blaCTX-M genes were co-harboured on conjugative plasmids with F2:A-:B- (n = 2), N (n = 1), F-:A-:B1 and N (n = 1) and untypable (n = 2) replicons. Both fosKP96-carrying plasmids belonged to replicon N. RFLP analysis showed that the two F2:A-:B- plasmids carrying fosA3 and blaCTX-M-3 genes shared the same pattern. Complete sequencing of one of the two F2:A-:B- plasmids, pFOS-HK151325 (69 768 bp) demonstrated it to be >99 % identical to the previously sequenced plasmid pHK23a originating from a pig E. coli isolate in the same region. This study demonstrated the dissemination of fosA3 genes in diverse E. coli clones on multiple blaCTX-M-carrying plasmid types, of which F2:A-:B- plasmids closely related to pHK23a were shared by isolates from human and animal sources.

Aims: To investigate plasmid-mediated fosfomycin resistance related to fosA3 in Escherichia coli isolates collected from different animals in Hong Kong, China, 2008-2010., Methods and Results: In total, 2106 faecal specimens from 210 cattle, 214 pigs, 460 chickens, 398 stray cats, 368 stray dogs and 456 wild rodents were cultured. The faecal colonization rates of fosfomycin-resistant E. coli were as follows: 11.2% in pigs, 8.6% in cattle, 7.3% in chickens, 2.4% in dogs, 0.8% in cats and 1.5% in rodents. The cultures yielded 1693 isolates of which 831 were extended-spectrum β-lactamases (ESBL) producers. Fosfomycin-resistant isolates were more likely than fosfomycin-susceptible isolates to be producers of ESBL and to have resistance to chloramphenicol, ciprofloxacin, cotrimoxazole, gentamicin and tetracycline. Of the 101 fosfomycin-resistant isolates, 97 (96.0%) isolates were fosA3 positive and 94 (93.1%) were bla(CTX) (-M) positive. PCR mapping showed that the fosA3-containing regions were flanked by IS26, both upstream and downstream in 81 (83.5%) isolates, and by an upstream bla(CTX-M-14) -containing transposon-like structure (ΔISEcp1-bla(CTX-M-14) -ΔIS903 or ISEcp1-IS10 -bla(CTX-M-14) -ΔIS903) and a downstream IS26 in 14 (14.4%) isolates. For the remaining two isolates, fosA3 was flanked by a downstream IS26 but the upstream part cannot be defined. In a random subset of 18 isolates, fosA3 was carried on transferable plasmids with sizes of 50-200 kb and the following replicons: F2:A-B- (n = 3), F16:A1:B- (n = 2), F24:A-B- (n = 1), N (n = 1), B/O (n = 1) and untypeable (n = 3)., Significance and Impact of the Study: This study demonstrates the emergence of fosA3-mediated fosfomycin resistance among multidrug-resistant E. coli isolates from various animals. IS26 transposon-like structures might be the main vehicles for dissemination of fosA3., (© 2012 The Society for Applied Microbiology.)

Few studies have compared CTX-M encoding plasmids identified in different ecological sources. This study aimed to analyze and compare the molecular epidemiology of plasmids encoding CTX-M-14 among strains from humans and animals. The CTX-M-14 encoding plasmids in 160 Escherichia coli isolates from animal faecal (14 pigs, 16 chickens, 12 cats, 8 cattle, 5 dogs and 3 rodents), human faecal (45 adults and 20 children) and human urine (37 adults) sources in 2002-2010 were characterized by molecular methods. The replicon types of the CTX-M-14 encoding plasmids were IncFII (n=61), I1-Iγ (n=24), other F types (n=23), B/O (n=10), K (n=6), N (n=3), A/C (n=1), HI1 (n=1), HI2 (n=1) and nontypeable (n=30). The genetic environment, ISEcp1 -bla(CTX-M-14) - IS903 was found in 89.7% (52/58), 87.7% (57/65) and 86.5% (32/37) of the animal faecal, human faecal and human urine isolates, respectively. Subtyping of the 61 IncFII incompatibility group plasmids by replicon sequence typing, plasmid PCR-restriction fragment length polymorphism and marker genes (yac, malB, eitA/eitC and parB/A) profiles showed that 31% (18/58), 30.6% (20/65) and 37.8% (14/37) of the plasmids originating from animal faecal, human faecal and human urine isolates, respectively, were pHK01-like. These 52 pHK01-like plasmids originated from diverse human (20 faecal isolates from 2002, 2007 to 2008, 14 urinary isolates from 2004) and animal (all faecal, 1 cattle, 1 chicken, 5 pigs, 9 cats, 1 dog, 1 rodent from 2008 to 2010) sources. In conclusion, this study highlights the importance of the IncFII group, pHK01-like plasmids in the dissemination of CTX-M-14 among isolates from diverse sources., (Copyright © 2012 Elsevier B.V. All rights reserved.)

This study assessed the temporal changes in the molecular epidemiology of bacteremic Escherichia coli isolates producing CTX-M-14 in Hong Kong. Blood isolates from 1996 to 1998 (period 1, n = 50) and 2007 to 2008 (period 2, n = 117) were investigated by molecular methods. CTX-M-type ESBL was carried by 98.2% (164/167) of the isolates. In both periods, the CTX-M-9 group and CTX-M-14 allele were the predominant ESBL type. The major clones were found to change from ST68 and ST405 in period 1 to ST131, ST69, and ST12 in period 2. Among 65 CTX-M-14-producing plasmids investigated further, 54 had the FII replicon. Replicon sequence typing and plasmid polymerase chain reaction-restriction fragment length polymorphism showed that 79.6% (43/54) of the FII plasmid subset was similar to the completely sequenced plasmid, pHK01 (human urine, Hong Kong, 2004). These pHK01-like plasmids were found to have spread to the major clones (ST68, ST405, and ST131) and multiple singleton isolates of all 4 phylogenetic groups., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objectives: To assess the occurrence of faecal carriage of Escherichia coli with resistance to 'critically important' antibiotics in various animals., Methods: Rectal or cloacal swabs were obtained weekly from cattle, pigs, chickens, cats, dogs and wild rodents over a 2 year period. Plain and antibiotic-containing medium was used for bacterial isolation. Selected isolates were characterized by molecular methods., Results: In total, 2106 faecal specimens from 398 cats, 460 chickens, 368 dogs, 210 cattle, 214 pigs and 456 rodents were cultured. The faecal carriage rate of extended-spectrum β-lactamase (ESBL)-producing E. coli was highest in pigs (63.6%, 136/214) and lowest in rodents (4.2%, 19/456). The faecal ESBL-producing E. coli carriage rate for food-producing animals (53.6%, 474/884) was significantly higher than that for cats/dogs (14.0%, 107/766; P<0.01) and wild rodents (4.2%, 19/456; P<0.01). ESBL-producing isolates from food animals often (33%-81%) had multidrug (≥4) resistance to amikacin, chloramphenicol, ciprofloxacin, co-trimoxazole, gentamicin, nalidixic acid, netilmicin, nitrofurantoin and tetracycline. Most (91.2%) of the ESBL-producing isolates had CTX-M-type enzymes. A total of 10 alleles (3, 13, 14, 15, 24, 27, 28, 55, 65 and 98) from two CTX-M families (M1 and M9) were found. PFGE showed that the CTX-M-producing isolates were genetically diverse., Conclusions: This study shows that food animals are a major reservoir of E. coli with multidrug resistance to many antibiotics that are ranked as critically important in human medicine.

A bacterial collection (n=249) obtained in Hong Kong from 2002 to 2004 was used to investigate the molecular epidemiology of aminoglycoside resistance among Escherichia coli isolates from humans and food-producing animals. Of these, 89 isolates were gentamicin-sensitive (human n=60, animal n=29) and 160 isolates were gentamicin-resistant (human n=107, animal n=53). Overall, 84.1% (90/107) and 75.5% (40/53) of the gentamicin-resistant isolates from human and animal sources, respectively, were found to possess the aacC2 gene. The aacC2 gene for 20 isolates (10 each for human and animal isolates) was sequenced. Two alleles were found that were equally distributed in human and animal isolates. PFGE showed that the gentamicin-resistant isolates exhibited diverse patterns with little clonality. In some isolates, the aacC2 gene was encoded on large transferable plasmids of multiple incompatibility groups (IncF, IncI1 and IncN). An IncFII plasmid of 140 kb in size was shared by one human and three animal isolates. In summary, this study showed that human and animal isolates share the same pool of resistance genes.

A prospective multicenter study was conducted to assess the epidemiology of antimicrobial resistance among uropathogens causing uncomplicated cystitis. Adult women with clinical diagnosis of uncomplicated cystitis were enrolled from 54 participating centers distributed all over Hong Kong during 2006 to 2008. A positive urine culture was found in 59.5% (352/592) patients. The patients had mean age of 44.9 years, and most (89.2%) were otherwise healthy. The most prevalent causative organism was Escherichia coli (77%), followed by other Enterobacteriaceae (14.2%), staphylococci (5.1%), and other Gram-positive bacteria (3.7%). The resistance rates of E. coli to co-trimoxazole and ciprofloxacin were 29.5% and 12.9%, respectively, and 14 isolates (5.2%) were confirmed as extended-spectrum beta-lactamase (ESBL) producers. Of the ESBL producers, molecular studies showed CTX-M-14, CTX-M-24, or CTX-M-9. Nitrofurantoin and fosfomycin were active against >90% of the isolates, regardless of resistance phenotypes for other drugs. Pulsed-field gel electrophoresis of representative isolates showed that the antibiotic-resistant strains were genetically diverse. Patients with history of recent antibiotic use were significantly more likely to have infection by E. coli with co-trimoxazole resistance (odds ratio [OR], 2.8; 95% confidence interval [CI], 1.4-5.7; P = 0.003) and ciprofloxacin resistance (OR, 2.5; 95% CI, 1.1-5.8; P = 0.03). Knowledge of the resistance data and risk factors could inform better use of antibiotics for empiric therapy for acute uncomplicated cystitis.

Aims: To compare the distribution of integrons and trimethoprim-sulfamethoxazole resistance genes among Escherichia coli isolates from humans and food-producing animals., Methods and Results: A collection of 174 multidrug-resistant E. coli isolates obtained from faecal samples of food-producing animals (n = 64) and humans (n = 59), and patients with urinary tract infections (n = 51) in Hong Kong during 2002-2004 were studied. The strains were analysed for their phylogenetic groups, the presence of sul genes (sul1 and sul2), integrons (intl1 and intl2) and class 1 integron-associated dfr cassette genes by PCR, restriction enzyme analysis and sequencing. Integrons were identified in 110 (63.2%) isolates. The prevalence of integrons was significantly different according to the specimen sources (animal faecal 84.4%, human faecal 67.8% and human urinary 31.4%) and phylogenetic groups (B1 80.8%, A 77.6%, D 54.1% and B2 11.5%). Faecal isolates (both human and animal) are more likely to belong to group A and B1. In contrast, most urinary isolates were either groups B2 and D. Among dfr containing isolates, dfrA1 and dfrA12 were almost exclusively found in strains of phylogenetic groups A and B1; and were present in animal and human faecal isolates. In contrast, dfrA17 was found in both faecal and urinary isolates and comprised strains from all phylogenetic groups. The sul1 and sul2 genes were equally prevalent among the isolates irrespective of the specimen source and phylogenetic group status. Pulsed-field gel electrophoresis analysis of isolates with identical cassette genes showed that they were genetically diverse., Conclusions: More animal faecal isolates carry class 1 integrons than human faecal and human urinary isolates, and the distribution of phylogenetic groups is common across animal and human faecal isolates but different from human urinary isolates., Significance and Impact of the Study: Commensal isolates from food-producing animals are an important reservoir for integrons carrying antibiotic resistance genes.