odong Xia

Presence and Characterization of Shiga Toxin-Producing Escherichia coli and Other Potentially Diarrheagenic E. coli Strains in Retail Meats

ABSTRACT To determine the presence of Shiga toxin-producing Escherichia coli (STEC) and other potentially diarrheagenic E. coli strains in retail meats, 7,258 E. coli isolates collected by the U.S. National Antimicrobial Resistance Monitoring System (NARMS) retail meat program from 2002 to 2007 were screened for Shiga toxin genes. In addition, 1,275 of the E. coli isolates recovered in 2006 were examined for virulence genes specific for other diarrheagenic E. coli strains. Seventeen isolates (16 from ground beef and 1 from a pork chop) were positive for stx genes, including 5 positive for both stx1 and stx2, 2 positive for stx1, and 10 positive for stx2. The 17 STEC strains belonged to 10 serotypes: O83:H8, O8:H16, O15:H16, O15:H17, O88:H38, ONT:H51, ONT:H2, ONT:H10, ONT:H7, and ONT:H46. None of the STEC isolates contained eae, whereas seven carried enterohemorrhagic E. coli (EHEC) hlyA. All except one STEC isolate exhibited toxic effects on Vero cells. DNA sequence analysis showed that the stx2 genes from five STEC isolates encoded mucus-activatable Stx2d. Subtyping of the 17 STEC isolates by pulsed-field gel electrophoresis (PFGE) yielded 14 distinct restriction patterns. Among the 1,275 isolates from 2006, 11 atypical enteropathogenic E. coli (EPEC) isolates were identified in addition to 3 STEC isolates. This study demonstrated that retail meats, mainly ground beef, were contaminated with diverse STEC strains. The presence of atypical EPEC strains in retail meat is also of concern due to their potential to cause human infections.

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.

Identification and antimicrobial resistance of extraintestinal pathogenic Escherichia coli from retail meats.

Extraintestinal pathogenic Escherichia coli (ExPEC) causes a variety of infections outside the gastrointestinal tract. Retail meats are frequently contaminated with E. coli strains, and they might serve as a vehicle for transmitting ExPEC. A total of 1,275 E. coli isolates recovered from ground beef, ground turkey, chicken breasts, and pork chops obtained in Georgia, Maryland, Oregon, and Tennessee in 2006 were investigated for the presence of ExPEC by using multiplex PCR. Identified ExPEC isolates were assigned to serogroups and phylogenetic groups and then analyzed for antimicrobial susceptibility. Approximately 16% (200 of 1,275) of the E. coli isolates were identified as ExPEC, based on defined genetic criteria. The occurrence of ExPEC was highest in E. coli isolated from ground turkey (23.5%) and chicken breasts (20.2%), and less frequent in isolates from pork chops (8.3%) and ground beef (3.4%). Phylogenetic grouping revealed that most (66.5%) ExPEC isolates fell into the same phylogenetic groups (B2 and D) as did virulent human ExPEC strains. Among the 15 antimicrobial agents tested, resistance to tetracycline (67.0%), sulfisoxazole (59.5%), and streptomycin (46.0%) was most frequent. Most ExPEC isolates (n = 163 [81.5%]) were resistant to at least one antimicrobial agent, and more than half (n = 114 [57%]) exhibited resistance to at least three drugs. This study found that ExPEC strains, including antimicrobial-resistant strains, were frequent among E. coli recovered from retail meats, especially those from chicken and turkey products. These findings indicate a need to better understand the role of certain meat types as potential sources of human ExPEC infection.

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.

Punicalagin Inhibits Salmonella Virulence Factors and Has Anti-Quorum-Sensing Potential

ABSTRACT Punicalagin, an essential component of pomegranate rind, has been demonstrated to possess antimicrobial activity against several food-borne pathogens, but its activity on the virulence of pathogens and its anti-quorum-sensing (anti-QS) potential have been rarely reported. This study investigated the efficacy of subinhibitory concentrations of punicalagin on Salmonella virulence factors and QS systems. A broth microdilution method was used to determine the MICs of punicalagin for 10 Salmonella strains. Motility assay and quantitative reverse transcription (RT)-PCR were performed to evaluate the effects of punicalagin on the virulence attributes and QS-related genes of Salmonella. The MICs of punicalagin for several Salmonella strains ranged from 250 to 1,000 μg/ml. Motility assays showed that punicalagin, at 1/16× MIC and 1/32× MIC, significantly decreased bacterial swimming and swarming motility, which corresponded to downregulation of the motility-related genes (fliA, fliY, fljB, flhC, and fimD) in RT-PCR assays. RT-PCR also revealed that punicalagin downregulated the expression of most of the selected genes involved in Salmonella virulence. Moreover, a QS inhibition assay indicated that punicalagin dose dependently inhibited the production of violacein by Chromobacterium violaceum and repressed the expression of QS-related genes (sdiA and srgE) in Salmonella. In addition, punicalagin significantly reduced Salmonella invasion of colonic cells (P < 0.01) with no impact on adhesion. These findings suggest that punicalagin has the potential to be developed as an alternative or supplemental agent for prevention of Salmonella infection.

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.

Tannin-Rich Fraction from Pomegranate Rind Damages Membrane of Listeria monocytogenes

Pomegranate rind has been reported to inhibit several foodborne pathogens, and its antimicrobial activity has been attributed mainly to its tannin fraction. This study aimed to investigate the antimicrobial activity of the tannin-rich fraction from pomegranate rind (TFPR) against Listeria monocytogenes and its mechanism of action. The tannin-related components of TFPR were analyzed by high-performance liquid chromatography and liquid chromatography-mass spectrometry, and the minimum inhibitory concentration (MIC) of TFPR was determined using the agar dilution method. Extracellular potassium concentration, the release of cell constituents, intra- and extracellular ATP concentrations, membrane potential, and intracellular pH (pHin) were measured to elucidate a possible antibacterial mechanism. Punicalagin (64.2%, g/g) and ellagic acid (3.1%, g/g) were detected in TFPR, and the MICs of TFPR were determined to be 1.25-5.0 mg/mL for different L. monocytogenes strains. Treatment with TFPR induced a decrease of the intracellular ATP concentration, an increase of the extracellular concentrations of potassium and ATP, and the release of cell constituents. A reduction of pHin and cell membrane hyperpolarization were observed after treatment. Electron microscopic observations showed that the cell membrane structures of L. monocytogenes were apparently impaired by TFPR. It is concluded that TFPR could destroy the integrity of the cell membrane of L. monocytogenes, leading to a loss of cell homeostasis. These findings indicate that TFPR has the potential to be used as a food preservative in order to control L. monocytogenes contamination in food and reduce the risk of listeriosis.

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.

Serotypes, virulence factors, and antimicrobial susceptibilities of vaginal and fecal isolates of Escherichia coli from giant pandas.

ABSTRACT Although Escherichia coli typically colonizes the intestinal tract and vagina of giant pandas, it has caused enteric and systemic disease in giant pandas and greatly impacts the health and survival of this endangered species. In order to understand the distribution and characteristics of E. coli from giant pandas, 67 fecal and 30 vaginal E. coli isolates from 21 giant pandas were characterized for O serogroups, phylogenetic groups, antimicrobial susceptibilities, and pulsed-field gel electrophoresis (PFGE) profiles. In addition, these isolates were tested for the presence of extraintestinal pathogenic E. coli (ExPEC) and diarrheagenic E. coli (DEC) by multiplex PCR detection of specific virulence genes. The most prevalent serogroups for all E. coli isolates were O88, O18, O167, O4, and O158. ExPEC isolates were detected mostly in vaginal samples, and DEC isolates were detected only in fecal samples. Phylogenetic group B1 predominated in fecal isolates, while groups B2 and D were frequently detected in vaginal isolates. Resistance to trimethoprim-sulfamethoxazole was most frequently observed, followed by resistance to nalidixic acid and tetracycline. All except five isolates were typeable by using XbaI and were categorized into 74 PFGE patterns. Our findings indicate that panda E. coli isolates exhibited antimicrobial resistance, and potentially pathogenic E. coli isolates were present in giant pandas. In addition, these E. coli isolates were genetically diverse. This study may provide helpful information for developing strategies in the future to control E. coli infections of giant pandas.

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.