Longitudinal study of prevalence of Campylobacter jejuni and Campylobacter coli from turkeys and swine grown in close proximity

Eastern North Carolina is a major contributor to both turkey and swine production in the United States. In this region, turkeys and swine are frequently grown in close proximity and by common growers. To further characterize colonization of turkeys and swine with Campylobacter in such a setting, we investigated the prevalence of thermophilic campylobacters in eight paired operations involving turkey farms in close proximity to finishing swine farms. All 15 surveyed flocks and 15 herds were Campylobacter positive at one or more sampling times. Campylobacter was isolated from 1,310 (87%) of the 1,512 turkey samples and 1,116 (77%) of the 1,448 swine samples. Most (> 99%) campylobacters from swine samples were Campylobacter coli, found in 59 to 97% of the samples from the different herds. Both Campylobacterjejuni and C. coli were recovered from the turkey flocks (overall prevalences of 52 and 35%, respectively). Prevalence among flocks ranged from 31 to 86% for C. jejuni and 0 to 67% for C. coli, and both species were recovered from most flocks. Relative prevalence of C. coli was higher in young birds (brooders), whereas C. jejuni predominated in grow-out birds (P < 0.0001). The prevalence of C. coli in a swine herd was generally not a good predictor for prevalence of this species in the corresponding turkey flock. These findings indicate that even though turkeys and swine grown in proximity to each other were commonly colonized with thermophilic campylobacters, the relative prevalences of C. jejuni and C. coli appear to be host associated.     

Distribution and prevalence of airborne microorganisms in three commercial poultry processing plants

Airborne microbial contaminants and indicator organisms were monitored within three poultry processing plants (plants A, B, and C). In total, 15 cubic feet (c.f.) of air was sampled per location during 15 visits to each plant and quantitatively analyzed for total mesophilic and psychrophilic aerobic counts, thermophilic campylobacters, Escherichia coli, and Enterobacteriaceae. The prevalence of Salmonella spp. in air samples was also evaluated. Significant reductions in total aerobic counts were observed between defeathering and evisceration areas of the three plants (P < 0.05). Mesophilic plate counts were highest in the defeathering areas of all plants compared to equivalent psychrophilic plate counts. Enterobacteriaceae counts were highest in the defeathering areas of all three plants with counts of log10 1.63, 1.53, and 1.18 CFU/15 c.f. recovered in plants A, B, and C, respectively. E. coli enumerated from air samples in the defeathering areas exhibited a similar trend to those obtained for Enterobacteriaceae with log10 1.67, 1.58, and 1.18 CFU for plants A, B, and C, respectively. Thermophilic campylobacters were most frequently isolated from samples in the defeathering areas followed by the evisceration areas. The highest mean counts of the organism were observed in plant A at 21 CFU/15 c.f. sample with plants B and C at 9 and 8 CFU/sample, respectively. With the exception of low levels of Enterobacteriaceae recovered from samples in the on-line air chill in plant A, E. coli, Enterobacteriaceae, or Campylobacter spp. were not isolated from samples in postevisceration sites in any of the plants examined. Salmonella spp. were not recovered from any samples during the course of the investigation.     

Antimicrobial resistance profile of five major food-borne pathogens isolated from beef, pork and poultry

This study was performed to evaluate the resistance rate against antimicrobials of food isolates of the five major food-borne pathogens to compare these and to possibly distinguish a pattern. A total of 922 samples of the major meat species (pork, beef and poultry) were analysed for thermophilic Campylobacter, Salmonella, Yersinia enterocolitica, pathogenic Escherichia coli and Listeria monocytogenes. Isolates were subjected to antimicrobial resistance testing by the disc diffusion method. Roughly the same overall rate of resistance was identified for thermophilic Campylobacter, Salmonella and pathogenic E. coli. Resistance to quinolones and tetracycline was determined most frequently. In contrast, food isolates of Y. enterocolitica and L. monocytogenes were rarely tested resistant. The significance of our findings is that resistance rates in enteric bacteria seem to be much higher than in pathogens found in a variety of environments, closely associated to the host environment.     

Novel Campylobacter isolation method using hydrophobic grid membrane filter and semisolid medium

Culture procedures for isolation of thermophilic campylobacters from food matrices are complex, labor intensive, and time-consuming. Most available methods include the use of antibiotics as selective agents to prevent the growth of competing microflora. A simple procedure for isolation of thermophilic campylobacters after enrichment in Rosef's enrichment broth was developed using a hydrophobic grid membrane filter (HGMF) on semisolid medium (SSM). SSM contains no antibiotics, and the HGMF physically separates Campylobacter from the enrichment broth, allowing isolation based on differential motility. The HGMF-SSM method was compared to the Agriculture and Agri-Food Canada Food Safety Procedures Manual (FSPM-10) method (Isolation of Thermophilic Campylobacters from Fresh Pork, Beef Veal, Poultry and Ready-to-Eat Meat Products), which includes the use of selective antibiotics. During the initial study, after enrichment the HGMF-SSM method yielded pure cultures of campylobacters after 16 to 18 h (overnight) compared with 48 h for the FSPM-10 method. Ninety-four turkey samples collected at local retail stores and 38 frozen pig fecal samples were processed by both methods. Thirty-five samples (26.5%) were positive by the HGMF-SSM method; 24 (18.2%) of these positive samples contained Campylobacter jejuni and 11 (8.3%) contained Campylobacter coli. With the FSPM-10 method, 25 samples (18.9%) were positive: 21 (15.9%) with C. jejuni and 4 (3%) with C. coli. For a subsequent field study, only the HGMF-SSM method was used to isolate Campylobacter from 1,200 chicken samples and 454 turkey samples sold at retail. Analysis of five subisolates from various samples indicated that only one type of Campylobacter was recovered by the HGMF-SSM method, as ascertained by MICs for 10 antimicrobials, sequencing of the short variable region of the flaA gene, and fingerprinting based on amplified fragment length polymorphism. The absence of antibiotics in the SSM may explain the higher recovery of thermophilic campylobacters. The HGMF-SSM method resulted in improved isolation of campylobacters and is simpler, faster, cheaper, and less labor intensive than the FSPM-10 method. The recovery of one type of Campylobacter from the chicken samples may have important implications, particularly in epidemiological studies.     

Occurrence and characterization of Listeria spp. in ready-to-eat retail foods from Vancouver, British Columbia

The occurrence of Listeria spp. and Listeria monocytogenes in retail RTE meat and fish products in Vancouver, British Columbia (B.C.) was investigated. To assess potential consumer health risk, recovered L. monocytogenes isolates were subjected to genotypic and phenotypic characterization. Conventional methods were used to recover Listeria spp. from deli meat (n = 40) and fish (n = 40) samples collected from 17 stores. Listeria spp. were recovered only from fish samples (20%); 5% harboured Listeria innocua, 5% had L. monocytogenes and 10% contained Listeria welshimeri. L. monocytogenes isolates serotyped as 1/2a and 1/2b, possessed dissimilar PFGE patterns, and had full-length InlA. Three 1/2a clonal isolates encoded the 50 kb genomic island, LGI1. Antimicrobial resistance (AMR) profiling showed all Listeria spp. possessed resistance to cefoxitin and nalidixic acid. L. monocytogenes were resistant to clindamycin, two were resistant to streptomycin, and one to amikacin. Reduced susceptibility to ciprofloxacin was seen in all L. monocytogenes, L. innocua and three L. welshimeri isolates. Reduced susceptibility to amikacin and chloramphenicol was also observed in one L. monocytogenes and three L. welshimeri isolates, respectively. Recovery of L. monocytogenes in fish samples possessing AMR, full-length InlA, LGI1, and serotypes frequently associated with listeriosis suggest B.C. consumers are exposed to high-risk strains.     

Incidence of Listeria spp. and Salmonella spp. in horsemeat for human consumption

The incidence of Salmonella spp., Listeria spp. and Listeria monocytogenes in horsemeat for human consumption was investigated. One-hundred and twenty-one samples of frozen horsemeat collected from two Brazilian abattoirs were analysed over a period of 1 year. Twenty-two samples (18.2%) were positive for Listeria spp. with nine (7.4%) containing L. monocytogenes. None of the samples harbored Salmonella spp.     

Radiation processing to ensure safety of minimally processed carrot (Daucus carota) and cucumber (Cucumis sativus): optimization of dose for the elimination of Salmonella Typhimurium and Listeria monocytogenes

Minimally processed vegetables are in demand, because they offer convenience to consumers. However, these products are often unsafe because of possible contamination with pathogens, such as Salmonella, Escherichia coli O157:H7, and Shigella species. Therefore, this study was carried out to optimize the radiation dose necessary to ensure the safety of precut carrot and cucumber. Decimal reduction doses (D-values) of Salmonella Typhimurium MTCC 98 were ca. 0.164 kGy in carrot samples and 0.178 kGy in cucumber samples. D-values of Listeria monocytogenes were determined to be 0.312 and 0.345 kGy in carrot and cucumber samples, respectively. Studies of inoculated, packaged, minimally processed carrot and cucumber samples showed that treatment with a 1-kGy dose of gamma radiation eliminated up to 4 log CFU/g of Salmonella Typhimurium and 3 log CFU/g of L. monocytogenes. However, treatment with a 2-kGy dose was necessary to eliminate these pathogens by 5 log CFU/g. Storage studies showed that both Salmonella Typhimurium and L. monocytogenes were able to grow at 10 degrees C in inoculated control samples. Neither of these pathogens could be recovered from radiation-processed samples after storage for up to 8 days.     

A note on the incidences of Salmonella spp., Listeria spp. and Escherichia coli O157:H7 serotypes in Turkish sausage (Soudjouck)

The incidence of Salmonella spp., Listeria monocytogenes and Escherichia coli O157:H7 was determined in 100 Turkish sausage (soudjouck) samples collected from shops and markets in the Afyon province, Turkey. Salmonella spp. were detected in 7% of the samples. All of the isolates were S. enterica Paratyphi B. In addition, Listeria spp. were detected in 9% of the samples. Its distribution was 7% L. monocytogenes and 1% each of L. ivanovii and L. innocua. Serological study of the seven L. monocytogenes isolates showed that three of these were 1/2 ab, three were 5/6 ab and one was 1 ab. E. coli O157:H7 was not detected in any of the samples. The pH values of the samples ranged from 4.8 to 6.5. In conclusion, increasing number of listeriosis and salmonellosis cases in Turkey and the contamination levels found indicate that risk assessment and improved preventive measures are required for these sausages.     

Lethality of Salmonella and Listeria innocua in fully cooked chicken breast meat products during postcook in-package pasteurization

The process lethality model was used to predict the thermal kill of Salmonella and Listeria innocua in fully cooked and vacuum-packaged chicken breast meat during hot-water postprocess pasteurization. Time-temperature profiles of the meat samples during treatment and D-values (decimal reduction times) and z-values (change in temperature required to change the D-value) for Salmonella and L. innocua in the same meat product were used in the prediction of lethality. The results of the model prediction were compared with those of the inoculation study for the same meat product at a 95% confidence level of up to 10(7) CFU/g for Salmonella and L. innocua. The thermal lethality predictions obtained with the process lethality model for Salmonella and L. innocua were within the 95% confidence level for the experimental data from the inoculation study, suggesting that the process lethality model was a useful tool for the determination of the kill of Salmonella or L. innocua at up to 10(7) CFU/g in fully cooked chicken breast meat products during postprocess pasteurization with hot water.     

A longitudinal study of Salmonella and Campylobacter jejuni isolates from day of hatch through processing by automated ribotyping

Comparisons of bacterial populations over long periods of time allow researchers to identify clonal populations, perhaps those responsible for contamination of farms or humans. Salmonella and Campylobacter can cause human illness, and our objective was to use a library typing system to track strains that persist in the poultry house and through the processing plant. Two farms, over four consecutive flocks, were studied. Multiple samples were taken of the poultry house environment, feed mill, transport crates, and carcasses in the processing plant. Sample collection on the farm took place on chick placement day, midgrowout, and the day of harvest. This study found that 80.3% of isolates belonged to a single strain of Salmonella Kentucky that persisted in several environmental samples for all flocks at both farms, from chick placement day to the final product at the plant. Surgical shoe covers produced most isolates (n = 26), and processing day yielded the highest recovery (n = 68). Additional serotypes were recovered, but the Salmonella Kentucky-positive eggshells and chick mortality appeared to be the source of the organism for both farms. All Campylobacter isolates recovered were identified as C. jejuni. Most Campylobacter isolates (90.1%) belonged to one of three core strains. C. jejuni was not recovered on chick placement day. Cecal droppings yielded all nine strains. Most isolates (98.2%) were from one farm. Cluster analysis grouped C. jejuni and Salmonella isolates into four and six distinct clusters, respectively, on the basis of a similarity level of 80%.     

The incidence of Salmonella and Listeria in raw milk from farm bulk tanks in England and Wales

Raw milks from farm bulk tanks in England and Wales were sampled over a 15 month period in 1992–93 and analysed for Salmonella spp, Listeria spp and Listeria monocytogenes. Of 1673 samples tested for Salmonella spp, six (0.36%) yielded a positive result. A total of 2009 samples were analysed for Listeria spp. Of these, 310 (15.43%) were found positive for Listeria spp, 102 (5.08%) of the positive samples yielded L monocytogenes, which represented 33% of the Listeria isolations. There was a significant rise in the isolation rate for listerias between October and March broadly in line with the period during which the cows were housed indoors. The highest isolation rate was in January, 25.89% in 1992 and 28.4% in 1993, and the lowest isolation rate (3.1%) in August. Although the incidence of L monocytogenes broadly paralleled the trend for total Listeria, the highest percentage of L monocytogenes to total Listeria was found in the months of April to September when the total Listeria isolations were at a minimum. Thirty two (1.59%) samples were found to be positive for Listeria spp prior to enrichment procedure, with 17 (0.85%) positive for L monocytogenes. The highest counts reported for L monocytogenes were 62 and 30 colony forming units (cfu)/ml with over 60% at a level of less than 10 cfu/ml. Of the direct isolations, some 78% were obtained between the months of March and July.     

A BELGIAN SURVEY OF HYGIENE INDICATOR BACTERIA AND PATHOGENIC BACTERIA IN RAW MILK AND DIRECT MARKETING OF RAW MILK FARM PRODUCTS

To monitor the bacteriological quality of raw milk and raw milk farm products, 143 samples of raw farm milk and 100 samples of raw milk farm products, 64 butters, 9 yogurts, 16 cheeses, 7 ice creams and 4 fresh cheeses, produced in Belgium were examined for coliforms, β-glucuronidase positive Escherichia coli, verotoxin producing Escherichia coli O157 , Staphylococcus aureus, Salmonella spp. , Listeria spp. and Listeria monocytogenes. The results were compared with the threshold and maximum values of the EC directive 92/46/EC or the maximum values of the Belgian Order of Council from September 3, 2000. The presence of staphylococcal enterotoxins was investigated on samples with S. aureus counts higher than the legal threshold values mentioned in the EC directive or, if not regulated in the directive, higher than the maximum value mentioned in the Belgian Order of Council. The obtained results for the hygiene-indicators coliforms, β-glucuronidase positive E. coli and S. aureus in the raw milk samples were comparable with most other industrialized countries. Compared to a prevalence of 0.7% and 6.3% for, respectively , E. coli O157 and L. monocytogenes, no Salmonella was found in the 25 g raw milk farm samples. The isolated E. coli O157 strain was confirmed to be verotoxigenic; it was positive for VT2 , eaeA and hly A. In butter not only a prevalence of 18.7% for L. monocytogenes in 25 g was found but also the maximum values for the hygiene-indicators mentioned in the Belgian Order of Council were often exceeded. No significant difference was found between the count of hygiene-indicators and the presence of Listeria spp. as well in raw milk as in raw milk butter. The bacteriological quality of on-farm made raw milk butter suggest that suitable hygienic conditions are not always provided. One of the 7 ice cream samples contained L. monocytogenes in 25 g.     

Occurrence of Listeria monocytogenes in freshwater fish farms and fish-smoking plants

Three Swiss fish farms, farming rainbow trout (Oncorhynchus mykiss), and their affiliated smoking plants were analyzed for the presence of Listeria spp. 590 samples were collected from the farming environment (raceway water, sludge), faecal content and skin of the fish, fish during processing, and the processing environment.Listeria spp. were found at prevalences of 2·3% in plant A, 31·6% in plant B (mainly L. monocytogenes), and 13·8% in plant C (mainly L. innocua). This high contamination rate in plant B may be explained by the following facts: (i) farm B uses river water flowing through agricultural land; (ii) plant B rears fish in earth ponds instead of concrete ponds or raceways; (iii) fish from farm B had not been denied feed prior to slaughter; and (iv) total lack of regular mechanical and chemical cleaning in the fish farm B and processing plant B.In all three plants samples taken after smoking but before packaging did not contain Listeria spp., although in plant B and C the raw fish was contaminated. Hygienic defaults during packaging can lead to contaminated ready-to-eat products, detected in plant B (L. monocytogenes) and plant C (L. innocua) with one sample each. To minimize a possible health hazard to the consumer, it is of great importance to prevent postprocessing contamination of smoked fish.Finally, means of preventing Listeria contamination during farming, slaughtering, processing and storage are suggested.     

High incidence of Listeria monocytogenes in European red smear cheese

The incidence of Listeria and Listeria monocytogenes in European red smear cheese was determined in order to assess whether the lack of recent outbreaks of listeriosis associated with cheese is due to improved hygenic conditions in the dairies. Out of European red-smear cheese samples of various types, 15.8% contained organisms of the genus Listeria, 6.4% of the samples were contaminated with L. monocytogenes, 10.6% with L. innocua, and 1.2% with L. seeligeri. Six cheese samples contained two or more Listeria species, including at least one L. monocytogenes isolate. The incidences of L. monocytogenes in cheeses from various countries were: Italy 17.4%, Germany 9.2%, Austria 10%, and France 3.3%. Listeria were found most frequently in soft and semi-soft cheese. Eight samples contained more than 100 L. monocytogenes cfu/cm2 cheese surface, 2 samples had counts above 10(4) cfu/cm2 cheese surface. Surprisingly, a higher incidence of L. monocytogenes was observed in cheeses made from pasteurized milk (8.0%) than in cheeses manufactured from raw milk (4.8%). Phage-typing of isolated Listeria strains clearly confirmed that (i) contaminations within dairy plants were persistent over a period of several weeks to months and (ii) that cross-contamination within the dairy plant is and important factor. Comparison of our data with past surveys seems to indicate that contamination of red smear soft cheese with L. monocytogenes has not decreased sufficiently over the past 15 years. It is therefore strongly recommended that these products are monitored carefully by cheese-making companies.     

The occurrence of enteric pathogens and Aeromonas species in organic vegetables.

A range of commercially available organic vegetables (n = 86) was examined for the presence of Salmonella, Campylobacter, Escherichia coli, E. coli O 157. Listeria and Aeromonas spp., to provide information on the occurrence of such organisms in organic vegetables in Northern Ireland. The study was not designed to quantify such organisms or to compare occurrence with conventionally farmed vegetables. Standard enrichment techniques were used to isolate and identify enteric pathogens and Aeromonas species. No Salmonella, Campylobacter, E. coli. E. coli O 157, Listeria were found in any of the samples examined. Aeromonas species were isolated from 34% of the total number of organic vegetables examined. Many (64%) of the organic vegetables examined were "ready-to-eat" after minimal processing, i.e., washing. Aeromonas spp. was isolated from 41% of these vegetables. Aeromonas spp. was not recovered from certain vegetable types. The most commonly isolated species of Aeromonas was Aeromonas schubertii with 21.0% of all samples contaminated with this species; 5.8% of samples contained A. hydrophila, 5.8% A. trota, 3.5% A. caviae and 2.3% contained A. veronii biovar veronii. Although Aeromonas species are frequently detected in organic vegetables, the absence of accepted enteric pathogens was encouraging, and does not support the allegation of organic foods being of high risk due to the farming methods used.     

Environmental dust exposure as a factor contributing to an increase in Escherichia coli O157 and Salmonella populations on cattle hides in feedyards

A study was conducted to determine the impact of exposure to dust in the cattle load-out area in feedyards on pathogen contamination of cattle hides. A total of 250 cattle hides were sampled during summer and fall months, which are associated with elevated prevalence of Escherichia coli O157 in West Texas. Animals were removed from their home pens and restrained in a chute and sampled prior to exposure to dust generated as a result of a simulated loading exercise. The cattle hides were sampled again after exposure to the loading dust to determine total numbers of pathogens on cattle hides on leaving their home pen (before loading) and on cattle hides after exposure to the dust in the loading area. Air and dirt samples from the home pens and the cattle load-out area were also collected. The presence of E. coli O157 and Salmonella was determined in all the samples, and when a positive sample was identified, the total numbers of these bacteria present were enumerated. The total numbers of pathogens increased after dust exposure; Salmonella counts increased from 1.09 log most probable number (MPN)/cm2 to 1.74 log MPN/cm2 after exposure, and E. coli O157 counts increased from 0.80 to 2.35 log MPN/cm2 after sampling. E. coli O157 and Salmonella were recovered from the air samples during dust generation at 6.66 and 11.1%, respectively. Salmonella and E. coli O157 prevalence was not changed and was not associated with the exposure to the dust. Results indicate airborne dust generated as a result of cattle movement and loading could be an important determining factor in total numbers of pathogens recovered on cattle hides.     

Microbiology of raw milk in New Zealand

The results of this study demonstrate the occurrence of the non-spore-forming pathogens, Staphylococcus aureus, Escherichia coli (total count and O157:H7), Listeria, Campylobacter and Salmonella, in New Zealand's raw milk supply. Samples of raw milk were collected monthly within five major dairying regions over one year. Each month, samples from five randomly selected farm vats in each region were collected for analysis (297 samples in total). Methods based on plate count techniques were used to enumerate S. aureus and E. coli. Enrichment methods in combination with a modified most probable number detection method were used to monitor samples for the presence of E. coli O157:H7, Listeria, Campylobacter and Salmonella. Salmonella was not detected in this study, and Campylobacter was isolated once (0.34%). E. coli was present at <100 cfu/ml in 99% of samples and exceeded 10(3)cfu/ml in 0.7% of samples. E. coli O157:H7 was not detected whereas non-pathogenic E. coli O157 strains (i.e. lacking genes for stx1, stx2, eae and Hly A) were detected in 1% of samples. S. aureus was not detected (<1 cfu/ml) in 21% of samples; levels were >1 but <100 cfu/ml in 60% of samples and on one occasion (0.34%) S. aureus exceeded 10(4)cfu/ml. L. monocytogenes was isolated from 0.68% of samples and L. innocua was present in 4% of samples. The results demonstrate that raw milk sampled from farm vats in New Zealand, as in other countries, inevitably contains recognised pathogens and, hence, control by pasteurisation or an equivalent treatment of raw milk remains paramount. Even so, the prevalence of most of these pathogens was lower than those reported in many of the studies performed in other countries.     

Prevalence of Listeria spp. in feces and carcasses at a lamb packing plant in Brazil

The objective of this work was to study the occurrence of Listeria species in feces and on dressed and cooled carcasses of lambs at a packing plant in Brazil. Listeria spp. were recovered on Oxford and Palcam agars. The 35 fecal samples yielded Listeria welshimeri (20%) and Listeria innocua (8.6%). The 69 carcass samples yielded L. innocua (34.8%), Listeria monocytogenes (4.3%), and Listeria ivanovii (1.5%). More Listeria spp. were recovered with two selective agars than with either agar alone.     

Low incidence of foodborne pathogens of concern in raw milk utilized for farmstead cheese production

Overall milk quality and prevalence of four target pathogens in raw milk destined for farmstead cheesemaking was examined. Raw milk samples were collected weekly from June to September 2006 from 11 farmstead cheese operations manufacturing raw milk cheese from cow's, goat's, and sheep's milk. Samples were screened for Listeria monocytogenes, Staphylococcus aureus, Salmonella, and Escherichia coli O157:H7 both quantitatively (direct plating) and qualitatively (PCR). Overall, 96.8% of samples had standard plate counts of < 100,000 CFU/ml, 42.7% of which were < 1,000 CFU/ml. Although no federal standards exist for coliforms in raw milk, 61% of samples tested conformed to pasteurized milk standards under the U.S. Pasteurized Milk Ordinance (PMO) at < 10 CFU/ml. All cow and sheep milk samples and 93.8% of goat milk samples were within the limits dictated by the PMO for somatic cell counts. Of the 11 farms, 8 (73%) produced samples that were positive for S. aureus, which was detected in 34.6% (46 of 133) of milk samples. L. monocytogenes was isolated from three milk samples (2.3%), two of which were from the same farm. E. coli O157:H7 was recovered from one sample of goat's milk for an overall incidence of 0.75%. Salmonella was not recovered from any of the 133 samples. The findings of this study suggest that most raw milk intended for farmstead cheesemaking is of high microbiological quality with a low incidence of pathogens. These data will help inform risk assessments associated with the microbiological safety of farmstead cheeses, particularly those manufactured from raw milk.     

Survey of retail alfalfa sprouts and mushrooms for the presence of Escherichia coil O157:H7, Salmonella,and Listeria with BAX,and evaluation of this polymerase chain reaction-based system with experimentally contaminated samples

BAX, a polymerase chain reaction (PCR)-based pathogen detection system, was used to survey retail sprouts and mushrooms for contamination with Escherichia coli O157:H7, Salmonella, Listeria spp., and Listeria monocytogenes. No Salmonella or E. coli O157:H7 was detected in the 202 mushroom and 206 alfalfa sprout samples screened. L. monocytogenes was detected in one sprout sample, and seven additional sprout samples tested positive for the genus Listeria. BAX also detected Listeria species in 17 of the mushroom samples. Only 6 of 850 PCR assays (0.7%) failed to amplify control DNA, and therefore reagent failures and the inhibition of PCR by plant compounds were rare. The sensitivity of the detection system was evaluated by assaying samples inoculated with 10 CFU of each of the pathogens. One hundred seventy-two alfalfa sprout samples were inoculated with E. coli O157:H7, and two sets of 130 samples were experimentally contaminated with Salmonella Enteritidis and L. monocytogenes. The frequency of detection depended on the protocols used for inoculation and culturing. Inoculation of samples with approximately 10 CFU from frozen stocks yielded detection rates of 87.5 and 94.5% for L. monocylogenes and Salmonella Enteritidis, respectively, in mushrooms. The corresponding rates for alfalfa sprouts were 94.5 and 76.3%. The E. coli O157:H7 detection rate was 100% for mushrooms but only 48.6% for sprouts when standard BAX culture protocols were used. The substitution of an overnight incubation in modified E. coli medium for the 3-h brain heart infusion incubation increased the rate of E. coli O157:H7 detection to 75% for experimentally contaminated sprouts. The detection rate was 100% when E. coli O157:H7 cells from a fresh overnight culture were used for the inoculation. Test sensitivity is therefore influenced by the type of produce involved and is probably related to the growth of pathogens in the resuscitation and enrichment media.