Meili Xi

Prevalence and characterization of Salmonella serovars in retail meats of marketplace in Shaanxi, China

A total of 764 retail meat including 515 chicken, 91 pork, 78 beef and 80 lamb samples were collected in Shaanxi Province of China in 2007-2008 to determine the prevalence of Salmonella. The isolates were characterized using serotyping, antimicrobial susceptibility testing, and the presence of bla(CMY-2) and bla(TEM) and class I integrons. Selective serovars were further subtyped using PFGE. Approximately 54% (276) of chicken, 31% (28) of pork, 17% (13) of beef and 20% (16) of lamb samples were positive of Salmonella. Among 24 serovars identified, Enteritidis (31.5%) was most common, followed by Typhimurium (13.4%), Shubra (10.0%), Indiana (9.7%), Derby (9.5%) and Djugu (7.0%). Nearly 80% of the isolates (283) were resistant to at least one antimicrobial, and 53% (191) to more than three antimicrobials. Resistance was most frequently observed to sulfamethoxazole (67%), to trimethoprim/sulfamethoxazole (58%) and to tetracycline (56%). Furthermore, many isolates were resistant to nalidixic acid (35%), ciprofloxacin (21%) and ceftriaxone (16%). Most isolates of Shubra (89%) and Indiana (88%) were resistant to > or = 9 antimicrobials, compared to only 11% of Enteritidis and 9% of Infantis that showed similar resistance. Class I integrons were detected in 10% of the isolates, and contained aadA, tetR, dhfr, bla(PSE-1), bla(DHA-1) and bla(VEB-1) gene cassettes alone or various combinations. Ceftriaxone- and/or cefoperazone-resistant isolates (n=62) carried bla(TEM) (51.6%) and/or bla(CMY-2) (56.5%). A total of 116 PFGE patterns were generated among 210 selected isolates. Our findings indicated that Salmonella contamination was common in retail meats, and that the Salmonella isolates were phenotypically and genetically diverse. Additionally, many Salmonella isolates were resistant to multiple antimicrobials.

Prevalence of Salmonella on Raw Poultry at Retail Markets in China

Data regarding Salmonella on raw poultry are very limited in China. The objective of this study was to determine the prevalence of Salmonella on raw poultry at the retail level in six provinces and two national cities in China. Whole chicken carcasses (n = 1,152) were collected from three types of retail markets (large, small, and wet). All samples were analyzed for the presence of Salmonella by using the U.S. Department of Agriculture, Food Safety Inspection Service method. Of 1,152 chicken samples, overall Salmonella prevalence was 52.2%. The highest prevalence was observed in Guangxi Province (65.3%), next in Guangdong Province (64.6%), and then in Beijing (63.9%), Shaanxi Province (50.7%), Henan Province (47.9%), Shanghai (44.4%), and Fujian Province (42.4%), and lowest prevalence was observed in Sichuan Province (38.9%). Salmonella prevalence was significantly different among the six provinces and two national cities. Salmonella prevalence was highest in the wet markets (54.4%) compared with the large markets (50.3%) and the small markets (52.1%), but differences were not significant (P > 0.05). Good manufacturing practices, good agricultural practices, and hazard analysis critical control point systems for Salmonella control in poultry production at the farm, processing, and retail level should be implemented.

Characterization of Staphylococcus aureus isolated from powdered infant formula milk and infant rice cereal in China.

Dry infant foods are not sterile and could be contaminated with various bacteria including certain pathogens. The aim of this study was to investigate the prevalence of Staphylococcus aureus in infant foods and to characterize these strains. A total of 367 infant food samples, including 143 samples of powdered infant formula milk (PIF) and 224 samples of infant rice cereal (IRC), were collected in the Shaanxi Province of China during the period of July to August 2010 and screened for S. aureus. All S. aureus isolates were characterized by antimicrobial susceptibility testing, pulsed-field gel electrophoresis (PFGE), and detection of genes encoding enterotoxins, exfoliative toxins, Panton-Valentine leukocidin (PVL), and toxic shock syndrome toxin 1. Among all the samples examined, sixteen of 143 PIF samples (11.2%) and 14 of 224 IRC samples (6.3%) were positive for S. aureus. From these positive samples, 29 S. aureus strains were isolated from PIF and 25 from IRC. Of these S. aureus isolates, 83.3% were resistant to at least one antimicrobial, 35.2% to three or more antimicrobials. Resistance was most frequently observed to erythromycin (75.9%), followed by ciprofloxacin (51.9%) and trimethoprim/sulfamethoxazole (27.8%), while significantly fewer isolates were resistant to gentamicin (22.2%), tetracycline (18.5%), or cefoxitin (3.7%). In addition, 63.0% of isolates were positive for one or more toxin genes tested. The three most predominant toxin genes were pvl (40.7%), seg (38.9%), and sec (18.5%), followed by sea (7.4%), seb (7.4%), sed (5.6%), and see (5.6%). The ets, tsst-1, seh, sei, and sej genes were not detected. A total of 39 PFGE patterns were generated among 51 selected food isolates. Our findings indicate that PIF and IRC in the Shaanxi province were contaminated with S. aureus, and many S. aureus isolates harbored multiple toxin genes and exhibited multiple antimicrobial resistance. In addition, these S. aureus isolates were genetically diverse. The presence of S. aureus strains in these infant foods poses a potential threat to infant health.

Counts, serotypes, and antimicrobial resistance of Salmonella isolates on retail raw poultry in the People's Republic of China.

The objective of this study was to determine Salmonella counts, serotypes, and antimicrobial resistance profiles in retail raw chicken meat in the Peoples Republic of China. Salmonella counts were determined according to the most-probable-number (MPN) method for 300 whole chicken carcasses. These samples were collected from large, small, and wet (open) markets in Guangdong, Shaanxi, and Sichuan provinces. Salmonella isolates were serotyped and tested for antimicrobial susceptibility. Of the 300 chicken carcasses, 43.3% were positive for Salmonella, with an overall mean of 1.7 log MPN per carcass (95% confidence interval, 1.6 to 1.8 log MPN per carcass). No significant differences (P > 0.05) were detected for storage temperature (i.e., chilled, frozen, or ambient), market type (large, small, or wet), province, or location (capital or noncapital city). Seventy-eight serotypes were identified among the 1,094 Salmonella isolates. The top five most common Salmonella serotypes on raw chicken carcasses were Enteritidis (19.2%), Indiana (15.2%), Typhimurium (14.6%), Agona (7.1%), and Thompson (6.6%). Salmonella isolates (n = 779) were most frequently resistant to sulfisoxazole (74.1%) and tetracycline (71.1%) and least resistant to ceftriaxone (22.5%) and cefoxitin (19%). Only 4% of the isolates were susceptible to all 15 antimicrobial agents, 45% were resistant to 1 to 5 agents, 29% were resistant to 6 to 10 agents, and 22% were resistant to 11 to 15 agents. Our findings revealed that Salmonella contamination was common in retail raw poultry in China, and the counts on contaminated carcasses were mostly low. Salmonella isolates were diverse in their serotype distribution and antimicrobial susceptibility profiles, with more than half of the isolates resistant to more than five antimicrobial agents. These data may be used in risk assessment models to reduce the transmission of Salmonella via chicken meat to humans in China.

Prevalence of extended-spectrum b-lactamase-producing Salmonella on retail chicken in six provinces and two national cities in the People's Republic of China.

Prevalence of extended-spectrum β-lactamase (ESBL)-producing Salmonella in food is not well documented. This study investigated the prevalence of ESBL-producing Salmonella in 699 Salmonella isolates recovered from 1,152 retail chickens collected from six provinces and two national cities in the Peoples Republic of China in 2011. ESBL-producing isolates were screened by double-disk synergy test and confirmed using PCR and DNA sequencing. Of the 699 isolates tested, 60 (8.58%) were identified to be ESBL-producing Salmonella. Prevalence of ESBL-producing Salmonella was the highest in Shanghai city (17 [24.64%] of 69), followed by Shaanxi (10 [15.38%] of 65), Fujian (9 [11.69%] of 77), Guangdong (9 [7.69%] of 117), Sichuan (5 [7.25%] of 69), Beijing (6 [5.17%] of 116), Henan (4 [4.65%] of 86), and Guangxi (0 [0%] of 100) province. Significant difference (P < 0.05) in the prevalence of ESBL-producing Salmonella was found among six provinces and two cities. No significant difference (P > 0.05) in the prevalence was found between wet markets and supermarkets or between whole chickens and chopped chickens. The prevalence of ESBL-producing Salmonella differed significantly (P < 0.05) among different seasons, being higher in autumn than in spring and winter. Overall, ESBL-producing Salmonella varied significantly (P < 0.05) among 12 detected Salmonella serotypes: Abony (1 [33.33%] of 3), Indiana (28 [28.57%] of 98), Edinburg (6 [24.00%] of 25), Shubra (2 [20.00%] of 10), Uppsala (1 [16.67%] of 6), Thompson (8 [14.81%] of 54), Haardt (1 [12.50%] of 8), Agona (3 [9.68%] of 31), Gueuletapee (1 [6.25%] of 16), Typhimurium (4 [5.56%] of 72), Heidelberg (1 [4.55%] of 22), and Enteritidis (4 [3.17%] of 126). This study revealed that ESBL-producing Salmonella do exist in retail chicken in the Peoples Republic of China and that the potential risk of their presence in foods needs further exploration.

Characterization of Extended-Spectrum β-Lactamase-Producing Escherichia coli Strains Isolated from Retail Foods in Shaanxi Province, China.

Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli strains have been reported worldwide; however, the incidence and characterization of foodborne ESBL-producing E. coli strains have been rarely reported in the Peoples Republic of China. Among a collection of 659 E. coli isolates recovered from retail foods in Shaanxi Province, Peoples Republic of China, 223 cefoxitin-resistant and/or cefoperazone-resistant isolates were screened for ESBL production with the double disk diffusion test. The ESBL-producing isolates were characterized for antimicrobial resistance and the presence of blaTEM, blaSHV, and blaCTX-M genes. Isolates with blaCTX-M were further classified by PCR as having blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, or blaCTX-M-25. One hundred forty-seven isolates were identified as ESBL positive. PCR detection revealed that 146 isolates (99.3%) contained the blaCTX-M gene. Among these isolates, 42 (28.8%) were positive for the enzyme CTX-M-1, 5 (3.4%) for CTX-M-2, and 99 (67.8%) for CTX-M-9. No CTX-M-8 and CTX-M-25 were found in this study. One hundred fifteen isolates (78.2%) were positive for the blaTEM gene, but blaSHV was not detected. Among the 147 ESBL-producing E. coli isolates, 75 (51.0%), 35 (23.8%), and 4 (2.7%) isolates were positive for blaTEM and blaCTX-M-9, blaTEM and blaCTX-M-1, and blaTEM and blaCTX-M-2, respectively. All of the 147 ESBL-producing isolates were resistant to three or more non-β-lactam antibiotics. This study provides evidence that foodborne E. coli can harbor ESBL-encoding genes. Thus, food could be a vehicle for the dissemination of ESBL-producing E. coli strains, a situation that requires surveillance and appropriate management strategies.

Antimicrobial susceptibility testing and genotypic characterization of Staphylococcus aureus from food and food animals.

Staphylococcus aureus is commonly present in humans and animals. The aim of this study was to investigate antimicrobial resistance and genetic characteristics of S. aureus from food and food animals in Shaanxi Province in China. A total of 332 nasal swabs, breast skin swabs, raw milk, and pork samples were collected from local pig, dairy farms, or local grocery stores and screened for the presence of S. aureus. S. aureus isolates were characterized using antimicrobial susceptibility, pulsed-field gel electrophoresis (PFGE) analysis, and polymerase chain reaction for detecting pvl and mecA genes. Methicillin-resistant S. aureus (MRSA) strains were additionally tested for SCCmec type and exfoliative toxin genes. The prevalence of S. aureus was 30.6% in pig nasal swabs, 32.5% in pork, 25.7% in cow nasal swabs, 30.8% in cow breast skin swabs, and 29.3% in milk samples. Resistances were common among isolates tested against erythromycin (65.7%), tetracycline (65.7%), ciprofloxacin (52.7%), followed by gentamicin (36.7%), chloramphenicol (23.1%), cefoxitin (8.3%), and oxacillin (7.7%), but no isolate was resistant to vancomycin, amikacin, or cefoperazone. pvl gene was found in the isolates from all types of samples except from cow nasal swabs. Fourteen isolates from pig nasal swabs contained mecA gene and were considered as MRSA. PFGE analysis showed that nasal isolates differed from food isolates, but isolates from the same animal source appeared to cluster closely. The PFGE patterns of MRSA isolates were different from other S. aureus isolates from pig nasal cavity even though they were from the same source. All the MRSA isolates belonged to SCCmec type IV(b). No isolates contained exfoliative toxin genes. These findings indicated that S. aureus, including multidrug-resistant S. aureus, are widely spread in food animals and animal-derived foods in Shaanxi Province, China. MRSA isolates from pigs may pose potential health risks for workers in swine farms and the community at large.

Prevalence and quantitative detection of Salmonella in retail raw chicken in Shaanxi, China.

Quantitative Salmonella concentrations and prevalence of Salmonella in raw chicken at the retail level in Shaanxi province, China, was determined in this study. Two hundred forty samples were collected in Yangling and the surrounding cities, in Shaanxi Province, China, for data collection over 12 successive months from April 2011 to March 2012. During the whole surveillance year, the overall Salmonella contamination rate of retail raw chicken was identified as 43.3%, the most-probable-number (MPN) values ranged from 0.0036 to 0.8596 MPN per g, and the average value was 0.1655 MPN per g, except in 11 of the samples, which had MPN values of more than 11 MPN per g. In April 2011, the highest prevalence rate, which was 80.0%, was observed, and the corresponding MPN value was 0.8596 MPN per g. Observed in November 2011, the lowest prevalence rate and the corresponding MPN value were 5.0% and 0.0036 MPN per g, respectively. Prevalence of Salmonella in chicken collected from the supermarket (49.2%) was higher but not significantly different from that in the wet markets (37.5%), although the MPN value of samples in the wet market (0.1912 MPN per g) was higher than that in the supermarket (0.1396 MPN per g). Prevalence of Salmonella was the highest in the frozen chicken (53.3%) compared with chilled chicken (45.0%) and freshly slaughtered chicken (37.5%); however, the MPN value of the freshly slaughtered chicken (0.1912 MPN per g) was higher than those of either frozen chicken (0.1804 MPN per g) or the chilled chicken (0.0988 MPN per g).

Emergence of β-Lactamases and Extended-Spectrum β-Lactamases (ESBLs) Producing Salmonella in Retail Raw Chicken in China

β-Lactamases and extended-spectrum β-lactamases (ESBLs) producing pathogenic bacteria were widely studied previously in China, but were seldom focused on foodborne Salmonella. In this study, an investigation concerning β-lactamases and ESBLs producing Salmonella recovered from retail raw chickens was performed. Sixty of 699 foodborne Salmonella isolates were detected as β-lactamases and ESBLs-producing ones that covered 12 Salmonella serotypes and exhibited different pulsed-field gel electrophoresis genotypes. Forty-four of 60 β-lactamases and ESBLs-producing strains were simultaneously resistant to ampicillin, amoxicillin/clavulanic acid, ceftiofur, ceftriaxone, and cefoxitin. The most commonly detected β-lactamases and ESBLs-encoding gene was bla(TEM-1) (n = 44), followed by bla(OXA-1) (n = 38), bla(CMY-2) (n = 29), bla(PSE-1-like) (n = 1), bla(CTX-M-3) (n = 16), and bla(CTX-M-15) (n = 1), respectively. Fourteen, 24, 21, and 1 isolates were detected simultaneously positive for 1, 2, 3, and 4 of the detected β-lactamases and ESBLs-encoding genes, respectively. A Salmonella strain simultaneously co-carrying bla(TEM-1), bla(OXA-1), bla(CMY-2), and bla(CTX-M-3) was first reported in the present study. Amino acid substitution of Trp244Cys/His247Leu was detected in PSE-1, Val218Asp in CMY-2, and Asp242Gly in CTX-M-15 enzymes, respectively. A difference was found among the amino acid sequences of the detected OXA-1, CMY-2, CTX-M, PSE-1, and TEM-1. The results demonstrated that β-lactamases and ESBLs were emerging and prevalent in foodborne Salmonella.

Distribution and Molecular Characterization of Salmonella enterica Hypermutators in Retail Food in China.

Hypermutable pathogens can easily acquire mutation opportunities, as well as antimicrobial resistance, and are tremendous hazards to food safety and public health. In this study, a total of 96 (7.6%) hypermutators were identified from 1,264 Salmonella isolates recovered from retail foods. Pulsed-field gel electrophoresis analysis indicated that hypermutators were genetically diverse. Amino acid substitution of Val421Phe was detected in MutS in one hypermutator and Val246Ala in 56 other hypermutators, while no mutation in MutS was found among the remaining 39 hypermutators. Hypermutators in Salmonella isolates recovered in 2010 (9.3%) and 2008 (7.7%) were significantly more prevalent than those in 2007 (1.4%). The rate of hypermutators in mutton (22.2%) was significantly higher than that in chicken (7.9%) and pork (4.7%). In Salmonella Leimo isolates (60.0%), hypermutators were most frequently detected, followed by Salmonella Essen (50.0%), Salmonella Indiana (36.6%), Salmonella Kallo (25.0%), Salmonella Heidelberg (23.8%), Salmonella Typhimurium (14.0%), Salmonella Shubra (13.0%), Salmonella Albany (11.1%), Salmonella Agona (7.0%), Salmonella Gueuletapee (6.3%), and Salmonella Enteritidis (1.7%). Salmonella hypermutators in isolates recovered from retail food stored at ambient temperature (15.7%) were significantly more prevalent than those stored in chilled (3.1%) and frozen (5.4%) condition. The overall distributions of mutation frequencies of the 96 hypermutators (selected by rifampin) were from 2.16 × 10(-5) to 4.25 × 10(-1). Mutation frequencies of hypermutators of Salmonella Leimo, Salmonella Essen, Salmonella Kallo, and Salmonella Agona were relative low, while those of Salmonella Typhimurium, Salmonella Indiana, and Salmonella Shubra were extremely high. No significant correlation was found between mutation frequency and antimicrobial resistance of the hypermutators.