Multiple antibiotic resistance index (MARI) of human-isolated Salmonella species: a practical bacterial antibiotic surveillance tool.

OBJECTIVES: Drug-resistant Salmonella plays a significant role in disease morbidity and mortality worldwide. The present study aimed to determine the multiple antibiotic resistance index (MARI) of Salmonella isolated from children hospitalized for gastroenteritis in Hong Kong. METHODS: Salmonella isolates from stool samples of children aged from 30 days to <5 years were confirmed by using MALDI-TOF MS and subjected to serotyping methods according to the White-Kauffmann-Le Minor scheme. Antimicrobial susceptibility was determined by agar disc diffusion. RESULTS: A total of 101 Salmonella isolates were serogrouped into Group B (n = 46, 45.5%), Group C (n = 9, 9.0%) or Group D (n = 46, 45.5%), and successfully classified into S. Enteritidis (n = 15) and S. Typhimurium (n = 7). Overall Salmonella susceptibilities demonstrated the highest level of resistance to ampicillin (76.2%), ciprofloxacin (54.0%) and tetracycline (61.2%) whereas MDR strains had high resistance toward ampicillin (100%), tetracycline (100%), cotrimoxazole (84.6%), chloramphenicol (83.3%) and ciprofloxacin (83.3%). MARI revealed that 80.2% of Salmonella including all MDR strains (n = 13) had indexes greater than 0.2. CONCLUSIONS: The MARI captures a snapshot of a high rate of antibiotic use and resistance in the isolated Salmonella, indicating the urgent need for continuous antimicrobial susceptibility surveillance and control of antibiotic prescription in selecting effective treatments for human diseases.

Genomic Epidemiology of Multidrug-Resistant Nontyphoidal Salmonella in Young Children Hospitalized for Gastroenteritis.

Nontyphoidal Salmonella (NTS) gastroenteritis in children remains a significant burden on health care and constitutes a majority of all admissions for Salmonella infections in public hospitals in Hong Kong. In this prospective study, 41% of 241 children hospitalized with gastroenteritis from three public hospitals during 2019 were culture confirmed to have NTS infection. These Salmonella isolates were whole-genome sequenced and in silico predicted for their serovars/serotypes using the Salmonella In Silico Typing Resource (SISTR) and SeqSero1, and the antimicrobial resistance (AMR) genes were determined. Phylogenetic analysis revealed three major clades belonging to Salmonella enterica serovar Enteritidis sequence type 11 (ST11) (43%), multidrug-resistant (MDR) S. Typhimurium ST19 (12%) and its monophasic variant ST34 (25%), and mostly singletons of 15 other serovars. MDR S. Typhimurium and its variant were more common in infants <24 months of age and possessed genotypic resistance to five antimicrobial agents, including ampicillin (A), chloramphenicol (C), aminoglycosides (Am), sulfonamides (Su), and tetracyclines (T). Older children were more often infected with S. Enteritidis, which possessed distinct genotypic resistance to AAmSu and fluoroquinolones. In addition, 3% of the isolates possessed extended-spectrum beta-lactamase (ESBL) CTX-M genes, while one isolate (1%) harboring the carbapenemase gene blaNDM-1 was identified. Our findings provide a more complete genomic epidemiological insight into NTS causing gastroenteritis and identify a wider spectrum of determinants of resistance to third-generation beta-lactams and carbapenems, which are often not readily recognized. With high rates of multidrug-resistant NTS from studies in the Asia-Pacific region, the rapid and reliable determination of serovars and resistance determinants using whole-genome sequencing (WGS) is invaluable for enhancing public health interventions for infection prevention and control. IMPORTANCE Nontyphoidal Salmonella (NTS) gastroenteritis is a foodborne disease with a large global burden. Antimicrobial resistance (AMR) among foodborne pathogens is an important public health concern, and multidrug-resistant (MDR) Salmonella is prevalent in Southeast Asia and China. Using whole-genome sequencing, this study highlights the relationship of the MDR Salmonella serotypes and the diverse range of Salmonella genotypes that contaminate our food sources and contribute to disease in this locality. The findings update our understanding of Salmonella epidemiology and associated MDR determinants to enhance the tracking of foodborne pathogens for public health and food safety.

Risk factors of non-typhoidal Salmonella gastroenteritis in hospitalised young children: a case-control study.

Objective: To explore risk factors associated with non-typhoidal Salmonella gastroenteritis in young children in Hong Kong. Design: A case-control study. Setting: Paediatrics wards at three public hospitals in Hong Kong. Participants: Cases were children aged above 30 days to below 5 years hospitalised for gastroenteritis at three public hospitals in Hong Kong with culture confirmed non-typhoidal Salmonella infection. Controls were age-matched (±2 months) children admitted for a reason other than gastroenteritis. Main outcomes measures: A face-to-face interview by using standardised questionnaire on exposures 3 days prior to illness. Adjusted OR (aORs) and 95% CIs were calculated using multivariable logistic regression. Results: A total of 102 cases and 204 age-matched controls were included in the analysis. Multivariable logistic regression revealed that having food purchased from places other than a supermarket, that is, from wet market/restaurant/farm (aOR, 2.64; 95% CI, 1.03 to 6.77; p=0.044) was a significant risk factor for non-typhoidal Salmonella infection. Having a household member with gastroenteritis symptoms (aOR, 2.03; 95% CI, 0.94 to 4.39; p=0.072) was of borderline significance and playing at a children's indoor playroom was a protective factor (aOR, 0.28; 95% CI, 0.09 to 0.85; p=0.024). Conclusions: Consumption of food purchased from places other than a supermarket was the identified determinant factor for non-typhoidal Salmonella gastroenteritis in Hong Kong. Parents/caregivers should be alerted to this risk when choosing foods for their young children. The protective effect of playing in an indoor playroom could be confounded by socioeconomic factors and further investigation is required to better understand its potential implication. There was some support for person-to-person transmission and good family hygiene needs to be emphasised.

Microorganisms as an Indicator of Hygiene Status Among Migrant Food Handlers in Peninsular Malaysia.

This study used microbial indicators to assess the hygiene status of 383 migrant food handlers from 3 urban cities in Peninsular Malaysia. Microbiological analysis revealed that all the hand swabs tested 99.5% positive for aerobic plate counts (mean [M] ± standard deviation [SD] = 3.57 ± 0.83 log10CFU [colony forming unit]), 20.8% positive for total coliform/ Escherichia coli (M ± SD = 0.30 ± 0.67 log10CFU), and 63.4% positive for Staphylococcus aureus (M ± SD = 1.38 ± 1.26 log10CFU). In addition, aerobic plate counts and Staphylococcus aureus counts exceeded the acceptable standard levels. Bacterial counts were found to be significantly associated with subjects' country of origin ( P = .019) and working responsibilities ( P = .001). Our findings indicate high probability of transmission of pathogenic bacteria from the food handlers' hands to customers during meal preparation and serving. This calls for improvements in personal hygiene and sanitation standards by the relevant health authorities among migrant food handlers.

Characterization of Nontyphoidal Salmonella Isolates from Asymptomatic Migrant Food Handlers in Peninsular Malaysia.

Asymptomatic Salmonella carriers who work as food handlers pose food safety and public health risks, particularly during food preparation, and this has serious implications for the disease burden in society. Therefore, we conducted a study to determine the number of Salmonella carriers in a migrant cohort in several food establishments in three major cities in Peninsular Malaysia. Sociodemographic data and stool samples were collected and analyzed using standard methods of detection and isolation. Antimicrobial susceptibility tests of the positive samples were also performed. A total of 317 migrant food handlers, originating from South and Southeast Asian countries, were recruited voluntarily. Nine (2.8%) stool samples were confirmed to be Salmonella positive. PCR serotyping and pulsed-field gel electrophoresis identified four serotypes as Typhimurium (n = 3), Corvallis (n = 2), Hadar (n = 1), Agona (n = 1) and two unknown serovars. Antimicrobial susceptibility tests revealed that all nine isolates were susceptible to amoxicillin-clavulanic acid, cefotaxime, ceftazidime, ceftriaxone, and gentamycin. However, seven isolates were found to be multidrug resistant to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, sulfonamides, streptomycin, and tetracycline. This study highlights that carriers of nontyphoidal Salmonella exist among migrant food handlers, which poses a health risk to consumers through food contamination. Our results indicate a need for authorities to enhance food safety awareness in the migrant workers and to reevaluate current health screening methods to include preventive measure such as mandatory stool screening as part of the preemployment and routine health examinations.

Pathogenic and saprophytic Leptospira species in water and soils from selected urban sites in peninsular Malaysia.

Leptospira species were studied in water and soils from selected urban sites in Malaysia. A total of 151 water (n=121) and soil (n=30) samples were collected from 12 recreational lakes and wet markets. All samples were filtered and inoculated into semi-solid Ellinghausen and McCullough modified by Johnson and Harris (EMJH) media supplemented with additional 5-fluorouracil. The cultures were then incubated at 30°C and observed under a dark field microscope with intervals of 10 days. A PCR assay targeting the rrs gene was used to confirm the genus Leptospira among the isolates. Subsequently, the pathogenic status of the isolates was determined using primer sets G1/G2 and Sapro1/Sapro2, which target the secY and rrs genes, respectively. The isolates were identified at serogroup level using the microscopic agglutination test (MAT) while their genetic diversity was assessed by pulsed field gel electrophoresis (PFGE). Based on dark field microscopy, 23.1% (28/121) water and 23.3% (7/30) soil cultures were positive for Leptospira spp. Of the 35 positive cultures, only 8 were pure and confirmed as Leptospira genus by PCR assay. Two out of 8 isolates were confirmed as pathogenic, 5 were saprophytic and one was intermediate. These 8 isolates were negative for the 25 reference hyperimmune rabbit sera tested in the MAT. PFGE showed that all 8 of these environmental Leptospira spp. were genetically diverse. In conclusion, the presence of pathogenic Leptospira spp. in the urban Malaysian environment may indicate and highlight the importance of water screening, especially in recreational lakes, in order to minimize any chance of Leptospira infection.