Eileen L. Lai

Fecal carriage of CTXM type extended-spectrum beta-lactamase-producing organisms by children and their household contacts.

OBJECTIVES To investigate the epidemiology of fecal carriage of CTX-M type extended-spectrum beta-lactamases (ESBL)-producing organisms among children and their household contacts. METHODS Fecal carriage with CTX-M-producing organisms was studied in 53 children and 172 household members. Molecular methods were used to characterize the isolates. RESULTS The children were mostly healthy and hospitalized for relatively mild febrile illnesses. Overall, the prevalence of fecal carriage of CTX-M-producing bacteria was 43.5% (admission children, 37.7%; household children, 20.7% and household adults, 50.3%). Household colonization index (defined by number of household carriers/total number of members) was significantly higher among families with at least one individual having a history of prolonged (>3 months) out-of-town residence in the previous year (mean+/-standard deviation; yes group, 0.67+/-0.36 vs. no group, 0.39+/-0.28, P=0.009) and was inversely correlated with the living space per person (R-square=0.139, P=0.006). Among 29 households with at least two carriers of CTX-M-producing enterobacteria, six clusters of clonally related strains were shared by 15 individuals from seven households; with both intra- and inter-household transmission. CONCLUSION CTX-M beta-lactamases may spread extensively amongst family members in the home.

Vancomycin MIC creep in MRSA isolates from 1997 to 2008 in a healthcare region in Hong Kong.

OBJECTIVES To assess whether vancomycin MIC creeps among blood methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from 5 hospitals in Hong Kong from 1997 to 2008. METHODS Blood cultures MRSA isolates from 1997 to 1999 (period 1), 2004 (period 2) and 2006-2008 (period 3) were retrieved. Etest method was used to determine their vancomycin MIC. The genotypic features were determined by PCR and sequencing. RESULTS 247 blood MRSA isolates were studied. The vancomycin MIC were 0.375, 0.5, 0.75 and 1 mg/L for 15 (6.1%), 68 (27.5%), 89 (36%) and 75 (30.4%) isolates, respectively. There was an increase in the percentage of isolates with an MIC=1mg/L from 10.4% (5/48) during period 1 to 21.6% (8/37) during period 2 and 38.3% (62/162) during period 3 (period 1 vs. period 3, P<0.001). Molecular typing showed that this was due to increased percentages of clonal cluster (CC) 8/SCCmec III/IIIA (agr group I), CC45/SCCmec IV/V (agr group IV) and other minor clones with elevated MIC over time. CONCLUSION This study found vancomycin MIC creep among blood MRSA isolates over time. As elevated MIC within the susceptible range may reduce vancomycin efficacy, clinical laboratories should adopt methods with the required precision to accurately determine MICs.

Prevalence and molecular epidemiology of plasmid-mediated fosfomycin resistance genes among blood and urinary Escherichia coli isolates

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.

Molecular epidemiology and nasal carriage of Staphylococcus aureus and methicillin-resistant S. aureus among young children attending day care centers and kindergartens in Hong Kong

OBJECTIVES To investigate the prevalence and molecular epidemiology of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) nasal carriage in children. METHODS We collected nasal and nasopharyngeal swabs from 2211 children aged 2-5 years attending 79 day care centers (DCCs) and 113 kindergartens (KGs) in all 18 geographical districts in Hong Kong. RESULTS The overall carriage rates of S. aureus and MRSA were 27.6% (95% confidence interval [CI], 24.8-28.5%) and 1.3% (95% CI, 0.8-1.8%), respectively. Molecular typing (staphylococcal cassette chromosome mec [SCCmec], sequence type [ST], clonal cluster [CC]) showed that all the 28 MRSA isolates had SCCmec IV (n = 13) or V (n = 15) including 12 isolates with community-associated-MRSA genotypes (ST59-IV/V, ST30-IV and ST88-V), 10 isolates with healthcare-associated-MRSA genotypes (ST45-IV/V, CC5-IV and ST630-V) and six isolates with novel genotypes (ST10-V and CC1-IV). Spa typing indicated that there was some within and between DCCs/KGs transmission of certain MRSA and methicillin-sensitive S. aureus strains but this was not extensive. CONCLUSION Our findings indicate the potential for DCCs to be a reservoir for emerging MRSA genotypes and highlight the need to enhance education and infection control measures to reduce their cross-transmission in this population.

Predominance of pHK01-like incompatibility group FII plasmids encoding CTX-M-14 among extended-spectrum beta-lactamase–producing Escherichia coli in Hong Kong, 1996–2008

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.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus in residential care homes for the elderly in Hong Kong

This territory-wide study evaluated the molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in residential care homes for elderly (RCHEs) in Hong Kong. MRSA colonization was assessed by taking swab culture from anterior nares and active skin lesions. Of 487 RCHEs surveyed, 80 MRSA strains were recovered from 1563 residents, giving a prevalence of 5.1%. Twenty-four isolates had SCCmec II, 2 had SCCmec III, 17 had SCCmec IV, 36 had SCCmec V, and 1 had a composite SCCmec type. Pulsed-field gel electrophoresis typing clustered 75 isolates into 7 clones (HKU10 to 50, HKU80, and HKU90). Two predominant types, HKU30 and HKU50, which together account for 75% of all MRSA strains, were found in 13 and 15 of the 18 geographic districts in Hong Kong, respectively. The main features for HKU50 strains were spa t1081/ST45-IV or V, capsular type 8, agrIV, and hla, seg, sei positive. On the other hand, HKU30 strains had spa t002/ST5-II, capsular type 5, agrII, and were hla, seg, sei positive. HKU30 strains were often multidrug resistant (MDR, involving ciprofloxacin, erythromycin, gentamicin, and tetracycline). In contrast, HKU50 strains exhibit both multidrug resistance (MDR) (involving ciprofloxacin, erythromycin, and tetracycline, but not gentamicin) and non-MDR patterns.

Changes in the epidemiology of methicillin-resistant Staphylococcus aureus associated with spread of the ST45 lineage in Hong Kong.

This study assessed the susceptibility trend of methicillin-resistant Staphylococcus aureus (MRSA) in 5 Hong Kong hospitals from 1995 to 2005. Representative blood isolates were characterized to correlate the changes in resistance phenotypes with the clonal nature of MRSA. The prevalence of multisusceptible (MS) MRSA, defined by sensitivity to gentamicin, co-trimoxazole, and fusidic acid, was found to increase dramatically from <2% in 1995 to 1997 to 4.5% to 5.8% in 1998 to 1999 and remained at around 10% thereafter. Isolation of MS-MRSA was significantly associated with older age, convalescent care, and blood culture source. Molecular typing showed that the increasing isolation of MS-MRSA was associated with the spread of ST45/Staphylococcus cassette chromosome mec (SCCmec) IV or V with spa t1768, t1857, or t1081. Other features of the ST45 strains are as follows: agr4, capsular type 8, and Panton-Valentine leukocidin (PVL) negative. Two other epidemic clones, EMRSA-16 (ST36/SCCmec II) and CC398 (ST1277/SCCmec V), were detected as sporadic isolates for the first time in Hong Kong.

Highly conjugative IncX4 plasmids carrying blaCTX-M in Escherichia coli from humans and food animals.

This study investigated the prevalence of IncX plasmid subtypes in commensal and pathogenic Escherichia coli isolates and the biological features of the IncX4 subtype. Two hundred and twenty-five E. coli isolates from multiple sources (47 chickens, 41 pigs, 30 cattle and 107 humans) obtained during the period 2006-2012 were tested for the presence of IncX1 to IncX5. Overall, the prevalence of IncX plasmids in chicken, pig, cattle and human isolates were 21.2 % (10/47), 19.5 % (8/41), 3.3 % (1/30) and 4.8 % (5/107), respectively. IncX4 was the most common subtype, followed by IncX1 and IncX3, while no IncX2 or IncX5 were found. Seven out of 16 (43.8 %) IncX4 plasmids were found to carry blaCTX-M genes and six of them originating from different host sources (four chickens, one pig and one human) had identical or highly similar RFLP patterns. Three IncX4 plasmids carrying blaCTX-M from different host sources were investigated further. It was found that the IncX4 plasmids had little effect on bacterial host growth parameters after their introduction to J53 recipients. Conjugation experiments demonstrated that the IncX4 plasmids could be efficiently transferred at 30-42 °C at rates which were generally 10(2)-10(5)-fold higher than those for the epidemic IncFII plasmid carrying blaCTX-M (pHK01). In conclusion, the IncX plasmids are more common than previously recognized. The efficient transfer of IncX4 plasmid at different temperatures and the lack of fitness burden on bacterial hosts highlight the ability of this plasmid replicon to be an important vehicle for dissemination of antimicrobial resistance.

Clonality and Antimicrobial Susceptibility of Staphylococcus aureus and Methicillin-Resistant S. aureus Isolates from Food Animals and Other Animals

ABSTRACT Out of 3,081 animals studied, 24.9% of pigs, 4.7% of chickens, 6.3% of dogs, 10.5% of cats, and 7.1% of rodents were Staphylococcus aureus positive. Prevalence of methicillin-resistant S. aureus (MRSA) was high in pigs (animals, 21.3%; batches, 46.5%), with all MRSA isolates and most methicillin-sensitive S. aureus isolates belonging to clonal complex 9 (CC9) and being multidrug resistant. The predominant S. aureus CCs among dog and cat isolates were similar. Among rodent isolates, CC398 predominated, with spa t034 the most frequent spa type detected.

Increase in the nasopharyngeal carriage of non-vaccine serogroup 15 Streptococcus pneumoniae after introduction of children pneumococcal conjugate vaccination in Hong Kong

This study assessed pneumococcal carriage in the early periods after routine use of 13-valent pneumococcal conjugate vaccine (PCV13) in Hong Kong. Nasopharyngeal swabs were obtained from 1110 children (<5 years) admitted with acute illness during September 2010-August 2013. Pneumococcal carriage rate was 13.5% in unvaccinated children, 14.1% in children who had ≥1 PCV dose and 15.3% in children who had ≥3 PCV doses. Nonv-PCV13 serotypes comprised 56.4% of all isolates. The most common serogroup/types were 15 (15A, 5.1%; 15B, 10.3%; 15C, 9.6%; 15F, 0.6%), 19F (17.9%), 6A (7.1%) and 6C (7.1%). Carriage of serogroup 15 was more common among vaccinated children (4.1% versus 0.6%, P = 0.033). Molecular typing revealed that expansion of several clones (clonal complex, CC63, CC199, CC1262, CC3397) was responsible for the increase in serogroup 15. Almost all CC63 and CC3397 isolates were nonsusceptible to both penicillin and erythromycin. The finding highlights the emergence of serogroup 15 following PCV13 use.