Determining the DNA Fingerprinting Profiles of Salmonella Isolates from Raw Poultry Meats and Human Clinical Samples from the Same Geographic Area Using Pulsed-Field Gel Electrophoresis.

ABSTRACT: Foods of animal origin, such as poultry, eggs, and pork, are recognized sources of Salmonella infections, but determination of the proportion of foodborne infections associated with various food sources has been challenging. In the present study, 141 Salmonella isolates recovered from 1,322 poultry product samples purchased over a 1-year period from retail stores across Seattle, WA were subtyped by pulsed-field gel electrophoresis (PFGE) using restriction enzyme XbaI. The objectives of the study were (i) to analyze the longitudinal distribution of Salmonella PFGE profiles throughout the sampling period and their clonality within and between poultry processing establishments, (ii) to determine the association between PFGE profiles of Salmonella isolates from locally distributed poultry products and those of clinical isolates submitted to the Washington State Department of Health (WA-DOH) laboratories, and (iii) to compare the PFGE profiles of Salmonella isolates from the National Antimicrobial Resistance Monitoring System (NARMS) retail meats program. During the 1-year sampling period, multiple indistinguishable PFGE patterns were found across multiple poultry processing establishments. Twelve of the 30 unique PFGE profiles of Salmonella isolates from locally purchased poultry products were indistinguishable from the PFGE profiles of clinical Salmonella isolates submitted to the WA-DOH. When the PFGE profiles from the poultry samples were compared with those found in the NARMS database, eight indistinguishable PFGE matches were found with isolates recovered from chicken breasts, ground turkey, and ground beef from multiple states. Although this study revealed some association between PFGE profiles from raw poultry products and those of clinical isolates from the same geographical area, these results do not prove that all of those clinical isolates were from infections acquired through consumption or handling of poultry.

Hospital-acquired listeriosis linked to a persistently contaminated milkshake machine.

One case of hospital-acquired listeriosis was linked to milkshakes produced in a commercial-grade shake freezer machine. This machine was found to be contaminated with a strain of Listeria monocytogenes epidemiologically and molecularly linked to a contaminated pasteurized, dairy-based ice cream product at the same hospital a year earlier, despite repeated cleaning and sanitizing. Healthcare facilities should be aware of the potential for prolonged Listeria contamination of food service equipment. In addition, healthcare providers should consider counselling persons who have an increased risk for Listeria infections regarding foods that have caused Listeria infections. The prevalence of persistent Listeria contamination of commercial-grade milkshake machines in healthcare facilities and the risk associated with serving dairy-based ice cream products to hospitalized patients at increased risk for invasive L. monocytogenes infections should be further evaluated.

Prevalence, concentrations, and antibiotic sensitivities of Salmonella serovars in poultry from retail establishments in Seattle, Washington.

Poultry have been identified as one of the major sources of salmonellosis, with estimates ranging from 10 to 22% of total cases. Despite several advances in the industry and new performance standards, the incidence of salmonellosis in the population has not declined over the last 15 years. Salmonella is pervasive in a wide variety of foods, and thus, estimating its burden resulting from specific food categories has been challenging and plagued with uncertainty due to critical data gaps. The objective of this study was to conduct a year-long market survey (1,322 samples) to help bridge the data gaps on the contamination rates and levels of Salmonella on raw poultry by product type (i.e., breast, thighs, drums, wings, and split breast) and production method (conventional versus organic). The isolates recovered were serotyped and tested for antibiotic sensitivities. A PCR method was utilized for initial screening of samples after an overnight enrichment in tryptic soy broth. Three-tube most-probable-number (MPN) assays and anti-Salmonella immunomagnetic separation methods were utilized to determine the levels of Salmonella and aid with the recovery of Salmonella species, respectively. Eleven percent of the samples were positive for Salmonella. Significant differences in percent positive rates by product type included up to a 4-fold difference in percent positive rates between establishments, ranging from 7 to 31%. Of the samples positive for Salmonella species, 94% had <30 MPN/100 g. Production methods identified as organic or as not using antibiotics had significantly higher rates of recovery of Salmonella. On the other hand, all of the Salmonella isolates that were resistant to two or more antibiotics originated from conventional processing establishments where antibiotics were utilized. In addition, a significant proportion of isolates from conventionally processed products were serotypes clinically relevant to humans.

Vancomycin-resistant Enterococcus spp. in marine environments from the West Coast of the USA.

AIMS: The aim of the study was to determine if vancomycin-resistant Enterococcus spp. [VRE] carrying vanA and/or vanB genes were present in public marine beaches and a fishing pier [2001-2003, 2008] from Washington and California [2008]. METHODS: PCR assays for the vanA and/or vanB genes with verification by DNA-DNA hybridization of the PCR products were used. Positive isolates were speciated using the BD BBL Crystal Identification and/or by sequencing the 16S ribosomal region. RESULTS: Eighteen (8%) of 227 isolates including Enterococcus faecalis, Enterococcus faecium, Enterococcus casseliflavus/gallinarum and a Staphylococcus epidermidis carrying vanA and/or vanB genes, from four of six Washington and one of two California sites, were identified. Selected VRE and the S. epidermidis were able to transfer their van genes to an E. faecalis recipient at frequencies ranging from 1.9 x 10(-6) to 6.7 x 10(-9). CONCLUSIONS: Vancomycin-resistant Enterococcus spp. was isolated from five of the seven sites suggesting that other North America public beaches could be the reservoirs for VRE and should be assessed. SIGNIFICANCE & IMPACT OF THE STUDY: This is the first report of isolation and characterization of VRE strains (and a vanB Staphylococcus sp.) from North American environmental sources suggesting that public beaches may be a reservoir for possible transmission of VRE to beach visitors.

Incidence of enterohemorrhagic Escherichia coli, Escherichia coli O157, Salmonella, and Listeria monocytogenes in retail fresh ground beef, sprouts, and mushrooms.

The objective of this study was to determine the prevalence of enterohemorrhagic Escherichia coli (EHEC), E. coli O157, Salmonella, and Listeria monocytogenes in retail food samples from Seattle, Wash. A total of 2,050 samples of ground beef (1,750 samples), mushrooms (100 samples), and sprouts (200 samples) were collected over a 12-month period and analyzed for the presence of these pathogens. PCR assays, followed by culture confirmation were used to determine the presence or absence of each organism. Of the 1,750 ground beef samples analyzed, 61 (3.5%) were positive for EHEC, and 20 (1.1%) of these were positive for E. coli O157. Salmonella was present in 67 (3.8%) of the 1,750 ground beef samples. Of 512 ground beef samples analyzed, 18 (3.5%) were positive for L. monocytogenes. EHEC was found in 12 (6.0%) of the 200 sprout samples, and 3 (1.5%) of these yielded E. coli O157. Of the 200 total sprout samples, 14 (7.0%) were positive for Salmonella and none were positive for L. monocytogenes. Among the 100 mushroom samples, 4 (4.0%) were positive for EHEC but none of these 4 samples were positive for E. coli O157. Salmonella was detected in 5 (5.0%) of the mushroom samples, and L. monocytogenes was found in 1 (1.0%) of the samples.

Prevalence of Shiga toxin-producing Escherichia coli in ground beef and cattle feces from King County, Washington.

Shiga toxin-producing Escherichia coli (STEC) is increasingly recognized as a common cause of diarrhea. STEC infection is a major public health threat because of its ability to cause serious and potentially life-threatening illnesses. The main reservoirs of STEC are believed to be the intestinal tracts of animals. Several studies have investigated the prevalence of STEC in various food items. The objective of this study was to determine the prevalence of STEC in the Seattle ground beef supply. In addition, the relative amount of STEC contamination between stores was compared, and possible differences between types of ground beef based on fat content (9, 16, and 23%) were investigated. A survey of Stx-I and/or Stx-II genes in fecal samples from cattle at a local slaughterhouse was also conducted. Of 296 ground beef samples tested from area retail grocery stores, 16.8% were positive for the presence of the toxin genes. Our data showed that there was no statistically significant difference (P > 0.05) in the prevalence of STEC between the ground beef samples of different fat contents and between grocery store chains. Of the 103 cattle fecal samples tested, 19 (18.4%) were found positive for the presence of Stx-I and/or Stx-II genes. The presence of a rather high percentage of STEC in the food supply in the absence of large number of cases suggests that not all STEC lineages are pathogenic for humans.

Effective and culturally acceptable water storage in Zimbabwe: maintaining the quality of water abstracted from upgraded family wells.

Because domestic water can be a vehicle of disease transmission in the home, there is a need for intervention. In Zimbabwe. 60 rural households obtaining water from shallow wells were selected for a field study. A water urn was designed, pretested, and field-tested. Thirty households designated as the case group were given two water urns each to substitute for traditional water storage containers (paint containers, oil containers, etc.). The remaining 30 households served as a control group. Samples were collected twice, at two-week intervals, from the water supply source (upgraded family wells) and storage containers (water urn or traditional containers) of each household (228 samples). Total coliform bacteria and fecal coliform bacteria were enumerated with the membrane filtration technique. A pretest of the water urn design showed a decline in turbidity that corresponded with a decline in bacterial counts. Wells supplying the case households had higher bacterial counts than those supplying the control households, but bacterial loads in the water urns were significantly lower than those observed in the corresponding supply wells (paired t-test: t = 3.97, df = 55, p < .01). Bacterial loads in traditional containers were similar to those observed in the supply wells (paired t-test; t = 0.2, df = 57, p > .05). The case group eagerly substituted water urns for traditional containers. The use of water urns was found to prevent or to reduce further contamination of well water after collection.

Recovery of Bordetella holmesii from patients with pertussis-like symptoms: use of pulsed-field gel electrophoresis to characterize circulating strains.

A 4-year retrospective study showing that we isolated Bordetella holmesii, but not Bordetella pertussis, from patients with pertussis-like symptoms was performed. From 1995 through 1998, we isolated B. holmesii from 32 nasopharyngeal specimens that had been submitted from patients suspected of having pertussis. Previously, B. holmesii had been associated mainly with septicemia and was not thought to be associated with respiratory illness. A study was undertaken to describe the characteristics of the B. holmesii isolates recovered and why we were successful in detecting the organism in nasopharyngeal specimens. B. holmesii isolates were characterized for drug sensitivities and for genetic relatedness by pulsed-field gel electrophoresis (PFGE). These isolates, an additional strain of B. holmesii isolated from a blood culture and previously confirmed by the Centers for Disease Control and Prevention, Atlanta, Ga., and 14 other clinical isolates of Bordetella spp., including 4 of B. bronchiseptica, 5 of B. parapertussis, and 5 of B. pertussis, were studied. They were all separately inoculated on three Bordet Gengou (BG) selective media containing either 0.625 microgram of oxacillin per ml, 40 microgram of cephalexin per ml, or 2.5 microgram of methicillin per ml, on BG agar with no antibiotic (control), and on charcoal agar (CA) with and without 40 microgram of cephalexin per ml. We found that cephalexin, the antibiotic commonly incorporated in both CA and BG agar for the recovery of Bordetella spp., is inhibitory to the growth of B. holmesii. In addition, the genotypic analysis of the 32 B. holmesii isolates by PFGE following restriction with XbaI and SpeI identified the dominant strains circulating during the study period.