Analysis of the upstream genetic structures of the ISEcp1-blaCTX-M transposition units in Escherichia coli isolates carrying blaCTX-M obtained from the Indonesian and Vietnamese communities.

Extended spectrum ß-lactamase (ESBL)-producing Escherichia coli have been found in healthy individuals in Indonesia and Vietnam. The ISEcp1-blaCTX-M transposition unit of ESBL-producing bacterial isolates has been considered responsible for the production of CTX-M type ESBL and it is important for the dissemination of blaCTX-M . This study aimed to characterize the upstream genetic structure (UGS) of E. coli isolates possessing blaCTX-M-1 group and/or blaCTX-M-9 group genes obtained from healthy individuals in Indonesia and Vietnam. A total of 501 CTX-M type ESBL-producing E. coli isolates possessing blaCTX-M-1 group and/or blaCTX-M-9 group genes were obtained from healthy individuals of the two countries in 2018. The UGSs of the ISEcp1-blaCTX-M transposition unit of the 501 ESBL-producing E. coli isolates were amplified by barcode-adaptor-ligation-mediated PCR and analyzed using the Nanopore sequencer. The obtained sequence information was used to classify the UGSs of the ISEcp1-blaCTX-M transposition unit. From the 501 ESBL-producing E. coli isolates, 502 UGSs were obtained, which were classified into 85 UGS types based on the sequence. ISEcp1 of 359 (71.5%) of the 502 UGSs was disrupted by gene insertion, and ISEcp1-blaCTX-M transposition unit of most (87.1%) of the determined UGSs was confirmed as plasmidic. Only 6 (7.1%) of the 85 UGS types were common to both countries. Our results indicated that many different UGSs of ISEcp1-blaCTX-M transposition units were detected in Indonesia and Vietnam; hence, we suggest that structurally different kinds of plasmids harboring blaCTX-M were separately distributed in the two countries.

A high-throughput sequencing determination method for upstream genetic structure (UGS) of ISEcp1-blaCTX-M transposition unit and application of the UGS to classification of bacterial isolates possessing blaCTX-M.

INTRODUCTION: Because blaCTX-M is responsible for resistance of bacteria to the third generation cephalosporins, location of blaCTX-M could be a good indicator for classifying bacterial isolates harboring blaCTX-M in molecular epidemiology. However, determination of blaCTX-M location has been difficult when multiple copies of ISEcp1 were found on bacterial genome. We aimed to establish a high-throughput analytical method for upstream genetic structures (UGS) of ISEcp1 to facilitate determination of blaCTX-M location. METHODS: Extracted DNA samples obtained from 168 Escherichia coli isolates possessing blaCTX-M were digested by restriction enzyme, HaeIII, and the digested DNA fragments were ligated with homemade barcode adaptors. Then, DNA fragments containing UGS of ISEcp1 were amplified and subjected to the Nanopore sequencer. RESULTS: Nucleotide sequences and locations of 168 UGSs obtained from the examined E. coli isolates were determined. Among the 168 determined UGSs, 150 (89.3%) UGS were confirmed on plasmid and classified into eight types. Interestingly, coding sequence of ISEcp1 transposase gene in seven of the eight types were disrupted by IS26 insertion. The remaining 18 (10.7%) UGSs were observed in identical chromosomal region. The obtained nucleotide sequences the locations of UGSs were confirmed by conventional capillary sequencer and Southern blotting, respectively, and any discrepant result was not observed with these confirmation procedures. CONCLUSIONS: Our results indicated that the established method was efficient for simultaneously determining at least 100 different UGS, and suggested that the determined UGSs of ISEcp1-blaCTX-M transposition unit was useful for classification of bacterial isolates harboring blaCTX-M.

Child stunting is associated with child, maternal, and environmental factors in Vietnam.

Child stunting in Vietnam has reduced substantially since the turn of the century but has remained relatively high for several years. We analysed data on children 6-59 months (n = 85,932) from the Vietnam Nutritional Surveillance System, a nationally representative cross-sectional survey. Multivariable Poisson regression models were used to estimate relative risk (RR) of stunting, stratified by child age and ecological region. Covariates at the child, maternal, household, and environmental levels were included based on available data and the World Health Organization conceptual framework on child stunting. Among children 6-23 months, the strongest associations with child stunting were child age in years (RR: 2.49; 95% CI [2.26, 2.73]), maternal height < 145 cm compared with ≥150 cm (RR: 2.04; 95% CI [1.85, 2.26]), living in the Northeast compared with the Southeast (RR: 2.01; 95% CI [1.69, 2.39]), no maternal education compared with a graduate education (RR: 1.77; 95% CI, [1.44, 2.16]), and birthweight < 2,500 g (RR: 1.75; 95% CI [1.55, 1.98]). For children 24-59 months, the strongest associations with child stunting were no maternal education compared with a graduate education (RR: 2.07; 95% CI [1.79, 2.40]), living in the Northeast compared with the Southeast (RR: 1.94; 95% CI [1.74, 2.16]), and maternal height < 145 cm compared with ≥150 cm (RR: 1.81; 95% CI [1.69, 1.94]). Targeted approaches that address the strongest stunting determinants among vulnerable populations are needed and discussed. Multifaceted approaches outside the health sector are also needed to reduce inequalities in socioeconomic status.

Carriage of Escherichia coli Producing CTX-M-Type Extended-Spectrum ß-Lactamase in Healthy Vietnamese Individuals.

Healthy carriage of CTX-M-type extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli was examined by thrice collecting fecal samples from the same 199 healthy Vietnamese subjects every 6 months. Using pulsed-field gel electrophoresis (PFGE), identical PFGE patterns throughout the three samplings were not observed, although prevalence of E. coli in the subjects was around 50% in the three samplings. Our results suggested a short carriage period of the CTX-M-type ESBL-producing E. coli in healthy Vietnamese subjects.

Potential transmission opportunity of CTX-M-producing Escherichia coli on a large-scale chicken farm in Vietnam.

OBJECTIVES: Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli have disseminated worldwide. This study investigated blaCTX-M-positive E. coli on a large-scale Vietnamese chicken farm and analysed whether there was any difference in prevalence and molecular characteristics of blaCTX-M-positive E. coli between the farm and the Vietnamese community. METHODS: Faecal samples were collected from 24 human individuals and 38 chickens from the large-scale chicken farm as well as 51 humans and 36 chickens from the community. All samples were collected between June 2013 and June 2014 in Bavi Province in the Red River Delta region of Vietnam. Molecular characterisation of CTX-M-producing E. coli and genetic relatedness among the isolates were evaluated by conventional typing methods. Antimicrobial susceptibility of the isolates was evaluated by the disk diffusion method. RESULTS: The prevalence of blaCTX-M-positive E. coli was 83.3%, 71.1%, 54.9% and 13.9% in farm workers, farm chickens, community individuals and community backyard chickens, respectively. On average, blaCTX-M-positive E. coli isolates from farm chickens were resistant to 8.3 different antibiotics. The average number of detected aminoglycoside-modifying enzyme genes (3.4 genes) and the detection rate of the plasmid-mediated colistin resistance gene mcr-1 (33.3%) were higher in blaCTX-M-positive E. coli isolated from farm chickens compared with other sampling groups. In addition, two types of indistinguishable pulsed-field gel electrophoresis (PFGE) patterns were observed in six blaCTX-M-65-positive E. coli and three blaCTX-M-55-positive E. coli from farm chickens. CONCLUSIONS: These results suggest a more frequent transmission opportunity of blaCTX-M-positive E. coli on the large-scale Vietnamese chicken farm.

Pre-germinated brown rice reduced both blood glucose concentration and body weight in Vietnamese women with impaired glucose tolerance.

We have reported that newly diagnosed type 2 diabetes mellitus (DM) patients in Vietnam have a low body mass index (BMI) of around 23 and that the major factor for this is high white rice (WR) intake. Brown rice (BR) is known to be beneficial in the control of blood glucose levels; however, it has the property of unpleasant palatability. Pre-germinated brown rice (PGBR) is slightly germinated by soaking BR in water as this reduces the hardness of BR and makes it easier to eat. This study was designed to evaluate the effect of a 4-mo PGBR administration on various parameters in Vietnamese women aged 45-65 y with impaired glucose tolerance (IGT). Sixty subjects were divided into a WR or PGBR group. For the first 2 wk, WR was replaced by 50% PGBR, then for 2 wk by 75% PGBR and from the second month 100%. Before the beginning of the study and at the end of the study, 1) anthropometric measurements, 2) a nutrition survey for 3 nonconsecutive days by the 24 h recall method and 3) blood biochemical examinations were conducted. Fasting plasma concentrations of glucose and lipids and the obesity-related measurements and blood pressure were favorably improved only in the PGBR diet group. The present results suggest that replacing WR with PGBR for 4 mo may be useful in controlling body weight as well as blood glucose and lipid levels in Vietnamese women with IGT.