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Salmonella is the most burdensome foodborne pathogen in the USA and a major causal agent of foodborne outbreaks. Detection of a pathogen such as Salmonella can be achieved within a few hours using commercially available rapid methods, but the sample preparation is time consuming and may require multiple days. We have developed and successfully tested an accelerated sample preparation method based on microfiltration, in some cases preceded by a short enrichment step, for the rapid detection of selected pathogens. The time-frame of the overall process, from sample preparation (i.e., food rinse or homogenate preparation, microbial enrichment, and filtration steps) to detection is 8 h or less. While microfiltration has been practiced for 70 years, the complex interactions between food substances and filter membrane surfaces have shown that food pretreatment methods need to be developed on a case by case basis for the recovery of bacteria from food homogenates and/or rinses. We have also demonstrated that addition of protease to treat homogenates of different poultry products prior to microfiltration avoids the rapid decrease in flux that otherwise occurs during microfiltration. This protease treatment minimizes filter clogging, so that the microbial concentration, recovery and detection of 1 to 10 CFU/g of Salmonella in poultry products is possible in less than 8 h.

Salmonella-contaminated foods, especially poultry-derived foods (eggs, chicken meat), are the major source of salmonellosis. Not only in the European Union (EU), but also in the United States, Japan, and other countries, has salmonellosis been an issue of concern for food safety control agencies. In 2005, EU regulation 1003/2005 set a target for the control and reduction of five target Salmonella enterica serovars-S. Typhimurium, S. Enteritidis, S. Infantis, S. Hadar, and S. Virchow-in breeding flocks. Thus, a simple biochip for the rapid detection of any of these five Salmonella serovars in poultry products may be required. The objectives of this study were to design S. Virchow-specific primers and to develop a biochip for the simultaneous identification of all or any of these five Salmonella serovars in poultry and poultry products. Experimentally, we designed novel polymerase chain reaction (PCR) primers for the specific detection of S. Virchow, S. Infantis, and S. Hadar. The specificity of all these primers and two known primer sets for S. Typhimurium and S. Enteritidis was then confirmed under the same PCR conditions using 57 target strains and 112 nontarget Salmonella strains as well as 103 non-Salmonella strains. Following multiplex PCR, strains of any of these five Salmonella serovars could be detected by a chromogenic biochip deployed with DNA probes specific to these five Salmonella serovars. In comparison with the multiplex PCR methods, the biochip assay could improve the detection limit of each of the Salmonella serovars from N×10 3 cfu/mL to N×10 2 cfu/mL sample in either the pure culture or the chicken meat samples. With an 8-hour enrichment step, the detection limit could reach up to N×10 0 cfu/mL., (Copyright © 2017. Published by Elsevier B.V.)

This observational study aims to investigate the microbiological quality of commercially prepared lightly cooked foods with a major component of food of animal origin and collected as would be served to a consumer. A total of 356 samples were collected from catering (92%), retail (7%) or producers (1%) and all were independent of known incidents of foodborne illness. Using standard methods, all samples were tested for: the presence of Campylobacter spp. and Salmonella spp. and enumerated for levels of, Bacillus spp. including B. cereus, Clostridium perfringens, Listeria spp. including L. monocytogenes, Staphylococcus aureus, Escherichia coli, Enterobacteriacea and aerobic colony count (ACC). Results were interpreted as unsatisfactory, borderline or satisfactory according to the Health Protection Agency guidelines for assessing the microbiological safety of ready-to-eat foods placed on the market. Amongst all samples, 70% were classified as satisfactory, 18% were borderline and 12% were of unsatisfactory microbiological quality. Amongst the unsatisfactory samples, six (2%) were potentially injurious to health due to the presence of: Salmonella spp. (one duck breast); Campylobacter spp. (two duck breast and one chicken liver pâté); L. monocytogenes at 4·3 × 103 cfu (colony-forming units)/g (one duck confit with foie gras ballotin) and C. perfringens at 2·5 × 105 cfu/g (one chicken liver pâté). The remaining unsatisfactory samples were due to high levels of indicator E. coli, Enterobacteriaceae or ACC.

One of the leading causes of foodborne illness in poultry products is Salmonella enterica. Salmonella hazards in poultry may be estimated and possible control methods modeled and evaluated through the use of quantitative microbiological risk assessment (QMRA) models and tools. From farm to table, there are many possible routes of Salmonella dissemination and contamination in poultry. From the time chicks are hatched through growth, transportation, processing, storage, preparation, and finally consumption, the product could be contaminated through exposure to different materials and sources. Examination of each step of the process is necessary as well as an examination of the overall picture to create effective countermeasures against contamination and prevent disease. QMRA simulation models can use either point estimates or probability distributions to examine variables such as Salmonella concentrations at retail or at any given point of processing to gain insight on the chance of illness due to Salmonella ingestion. For modeling Salmonella risk in poultry, it is important to look at variables such as Salmonella transfer and cross contamination during processing. QMRA results may be useful for the identification and control of critical sources of Salmonella contamination.

Aims: To determine the effect of hygiene measures on cross-contamination of Campylobacter jejuni at home and to select a safe tracer organism for C. jejuni., Methods and Results: Comparative tests were conducted with nonpathogenic Escherichia coli and Lactobacillus casei and L. casei was chosen as the safe tracer organism. Salads containing chicken breast fillet contaminated with a known number of C. jejuni and L. casei were prepared according to different cross-contamination scenarios and contamination levels of salads were determined. Cross-contamination could be strongly reduced when cleaning cutting board and cutlery with hot water (68 degrees C), but generally was not prevented using consumer-style cleaning methods for hands and cutting board., Conclusions: Dish-washing does not sufficiently prevent cross-contamination, thus different cutting boards for raw meat and other ingredients should be used and meat-hand contact should be avoided or hands should be thoroughly cleaned with soap. Lactobacillus casei can be used as a safe tracer organism for C. jejuni in consumer observational studies., Significance and Impact of the Study: Cross-contamination plays an important role in the transmission of food-borne illness, especially for C. jejuni. This study delivers suitable data to quantitatively assess the risk of campylobacteriosis caused by cross-contamination and it shows the effect of different preventive hygiene measures.

Changes in food production and societal pressures have led to a continuing increase in the incidence of foodborne illness. Many pathogens are associated with specific foods, e.g., E. coli O157:H7 with hamburgers or Salmonella with eggs. The U.S. Food and Drug Administration has recently approved irradiation for sterilization of meat, but public acceptance of irradiated food is low. Because contaminated foods are seldom detected before they reach store shelves, care in food preparation by professional and home cooks is crucial.

Survival and growth of Listeria monocytogenes isolates during sous vide processing and storage, and the applicability of predictive modelling in determining the potential for growth of L. monocytogenes in broth models and in sous vide products was investigated. L. monocytogenes grew in anaerobic tryptose phosphate broth and in chicken and beef samples by 2 log cycles in 8 days at 3 degrees C and 4-5 log cycles in 6 days at 8 degrees C. However, heating to an internal temperature of 70 degrees C resulted in a 4-5 log reduction and 70 degrees C/2 min resulted in a reduction greater than 7 log cycles. Lowering the product pH to 5.0 was effective in inhibiting L. monocytogenes growth, whereas a sodium chloride concentration of 2% had a negligible effect on growth rates. The square root model (Ratkowsky et al., 1983) predicted L. monocytogenes growth rates at 0-25 degrees C with a coefficient of determination (R2 value) of 98.36-99.63% and a bias factor of 1.08 to 1.21 in beef, chicken and broth substrates of unmodified pH. In addition, the Response Surface Polynomial Model (Version 3.1, Buchanan et al., 1989) predicted generation times at 5-25 degrees C with a 0-17.4% difference between observed and expected generation times in tryptose phosphate broth at pH 7.3. There were however, large differences (25.5 vs. 5.3 h) between observed generation times at pH 5.6 (8 degrees C) and those predicted by the Pathogen Modelling Program in tryptose phosphate broth. A divergence from predicted values was also noted at lower temperatures (0-3.5 degrees C) in the square root model.

Microbiological contamination of foods and drinking water is a global problem, and a significant amount of expense is being incurred as a result of such contamination. The microorganisms associated with almost half of all disease outbreaks still go unidentified, primarily as a result of inadequate monitoring and surveillance. Though significant improvements have been made in refining molecular methods for detecting infectious agents, a majority of these methods are being employed only on clinical samples where pathogen densities are much higher than those found in environmental and food samples. Comparative evaluations of the various protocols in terms of cost, sensitivity, specificity, speed, and reproducibility need to be undertaken so that the true applicability of these methods can determined. In the future, molecular methods, especially gene amplifications and in situ hybridizations, will find increasing applications in the differentiation of viable and non-viable organisms, in predicting antimicrobial resistance, and in the identification and characterization of unculturable microorganisms. Though molecular detection methods will not totally replace conventional methods, they will significantly enhance our ability to detect microbial pathogens rapidly.

Eight hundred and eighty-three strains of Campylobacter spp. isolated between 1982 and 1989 from human stools and poultry products were screened for quinolone resistance. In this period the prevalence of resistant strains isolated from poultry products increased from 0% to 14%. During the same period the prevalence in man increased from 0% to 11%. The emergence of quinolone resistance has implications for the identification of campylobacter up to species level: the susceptibility for nalidixic acid can no longer be used as a criterion for identification in the laboratory. The rapid emergence of resistant campylobacter may also have important implications for the treatment and prophylaxis of diarrhoeal disease. The increase of quinolone resistance coincides with the increasing use of fluoroquinolones in human and veterinary medicine. Extensive use of enrofloxacin in poultry and the almost exclusive transmission route of campylobacter from chicken to man, in The Netherlands, suggests that the resistance observed is mainly due to the use of enrofloxacin in the poultry industry.

A direct selective enrichment procedure was developed for the isolation of Campylobacter jejuni from poultry products. The selective enrichment medium (ATB) consisted of (per liter) tryptose (20 g), yeast extract (2.5 g), sodium chloride (5 g), FBP supplement (ferrous sulfate [0.25 g], sodium metabisulfite [0.25 g], sodium pyruvate [0.25 g]), bicine (10 g), and agar (1 g). Hematin solution (6.25 ml; prepared by dissolving 0.032 g of bovine hemin in 10 ml of 0.15 N sodium hydroxide solution and autoclaving at 0.35 kg/cm2 for 30 min), rifampin (25 mg), cefsulodin (6.25 mg), and polymyxin B sulfate (20,000 IU) were added after the medium was sterilized. The pH was adjusted to 8.0. Samples were enriched in the above medium at 42 degrees C for 48 h under an atmosphere of 5% O2, 10% CO2, and 85% N2. Enrichment cultures were streaked on a plating medium composed of Brucella agar, hematin solution, FBP supplement, and the above antibiotics. Plates were incubated under the same conditions as above. Suspect colonies from the plates were confirmed to be C. jejuni by morphological examination, growth characteristics, and biochemical tests. The above method yielded 25 isolates of C. jejuni from 50 samples of retail cut-up chicken and chicken parts, whereas a more complex method involving filtration, centrifugation, selective enrichment under a flowing atmosphere, and membrane filtration yielded only 6 positives from the same samples. The new isolation procedure was particularly effective in isolating C. jejuni in the presence of large numbers of Pseudomonas aeruginosa.(ABSTRACT TRUNCATED AT 250 WORDS)

A collaborative study was performed in 13 laboratories to validate an enzyme immunoassay (EIA) procedure for rapid detection of Salmonella in foods. The EIA was compared with the standard culture procedure for detection of Salmonella in 6 food types: ground black pepper, soy flour, dried whole eggs, milk chocolate, nonfat dry milk, and raw deboned turkey. Uninoculated and inoculated samples were included in each food group analyzed. There was no significant difference in the proportion of samples positive by the EIA and culture procedures at the 5% level for any of the 6 foods. The enzyme immunoassay screening method has been adopted official first action as a rapid screening method for detection of Salmonella.

Summarized results of five-year microbiologic studies on the production of dried egg products (white of egg, yolk, and melange), are presented. Studied were a total of 1402 batches of dried products, 231 samples of raw material, and 2348 samples of dried semi-products. The effect was followed up of the technologic processes and the hygiene status on the quantitative and the qualitative composition of the residual microflora. It was found that dried egg products met to a large extent the requirements of the microbiologic standards of the hygiene state and the epidemiologic safety and could be used on a mass scale in the food industry.

Between the period of August 1973 and May 1974, 553 roasting chickens, soup chickens, ducks, and geese from seven different countries were examined for the presence of Salmonella. In 284 (= 51,4%) specimens 23 various Salmonella serotypes were discovered. Out of 475 Salmonella isolates 244 (= 51,4%) were resistant to streptomycin, the sulfonamide complex or their combination whereas 195 (= 41,0%) were sensitive to all eight tested antibiotics. Of greater importance for nutritive and therapeutic application of antibiotics is, however, the resistance of 36 (= 7,6%) Salmonella strains to tetracyclines, ampicillin and kanamycin, 28 (= 5,9%) of the salmonellae showing resistance exclusively to tetracyclines. The water tested from 10 thawed specimens 2 of which contained diphosphates, produced dubious results with the general inhibitor test. Regarding the bacterial yield, both the selenite and tetrathionate enrichments were of equal value. Nevertheless with each individual enrichment the isolated partial amount of the entire collection of positive samples was not identical so that the highest yield of Salmonella was obtained from a combination of both methods. Incubation of the selenite enrichment at a maximum of 43 degrees C definitely produced a higher percentage than at 37 degrees C. A selenite enrichment proved to be superior to that of tetrathionate for the isolation of Salmonella serotypes which rarely occur in fowl. The excellent selectivity of the Brilliant-green Phenol-red Lactose Sucrose Agar, as repeatedly described in the literature, is also confirmed by these tests.

During a Salmonella survey in a large integrated poultry organization it was observed that a significant correlation existed between Salmonella serotypes isolated from the raw feed ingredients and those from finished carcases. A number of serotypes hitherto unrecognized in the organization were detected in the raw feed ingredients, and were later recognized in live birds and carcases from the processing plant. It appears that a significant reduction in carcase contamination rate could be achieved by minimizing Salmonella in the meal and grain constituents of poultry feed.

1. Heat destruction of bacteria attached to the skin did not occur at a logarithmic rate provided the temperature was above 51 degrees C. 2. It was concluded that the bacteria are protected by their location in the skin surface rather than by polymers produced during attachment. 3. An analogous process is considered to take place during the scalding of broilers, since bacteria which survived were difficult to remove from the carcasses during further processing.

Two brands of frozen fried chicken products were purchased monthly from a local supermarket for six months. Microbiological qualities of these samples were studied. The log number of mesophilic counts ranged from 2.90/g. to 4.78/g; log psychrophilic counts varied from 2.74/g. to 4.66/g.; and log Staphylococcus- 110 medium counts ranged from 2.84/g. to 4.54/g. Mean log microbial counts were higher in brand A samples than in brand B samples. No yeast or mold was detected and all samples were Salmonella negative. Most samples were negative in coliform test, except five samples had coliform MPN ranging from 0.5/g to 0.8/g. A total of 144 isolates of psychrophiles from 12 samples was tentatively identified to be members of Staphylococcus species. About 82.2% of the isolates belong to the Subgroup VI Staphylococcus according to Baird-Parker's classification. Another 17.8% of the isolates resembled Subgroup vi Staphylococcus except in phosphatase test.

A study was made in order to improve a new Salmonella identification test (Mucap Test) in which umbelliferone is released, giving a blue fluorescent light under a Wood lamp, after contact with Salmonella colonies. The study concerned 354 colonies, previously isolated from 55 poultry meat samples. Two enrichment media [Tetrathionate Bile Broth (TBB) and Rappaport Vassiliadis (RV)] and two isolation media [Brilliant Green Agar (BGA) and Desoxycholate Agar (DA)] were used, and the results of the test obtained respectively with each association were compared. The sensitivity was consistently good, but the specificity of the test was generally poor. The best association seemed to be RV/DA which gave 85% specificity, against 39% for TBB/BGA, 58% for TBB/DA, and 77% for RV/BGA. The predominant genera responsible for false-positive results were Pseudomonas and Proteus Providencia.

A collaborative study was performed to validate the performance of the 1-2 TEST for detection of motile salmonellae in foods. Detection is based on observation of an immobilized band of cells. Twenty-three laboratories participated in the study. The 1-2 TEST (immunodiffusion test) was compared with the standard culture procedure (BAM/AOAC; FDA Bacteriological Analytical Manual) for detection of Salmonella in 6 food types: ground black pepper, soy flour, dried whole egg, milk chocolate, nonfat dry milk, and raw deboned turkey. Uninoculated and inoculated samples were included in each food group analyzed. After the tests on the 6 foods were completed, analysis of the data for turkey and soy flour showed that certain collaborators obtained data inconsistent with the data from the majority of collaborators. No specific method deviations to account for the inconsistencies were reported by those collaborators, so the collaborative testing of these 2 foods was repeated. Analysis of data for pepper, chocolate, nonfat dry milk, dried whole egg, and the second set of soy flour and turkey indicated 96.1% agreement between the BAM/AOAC and immunodiffusion test methods. The false negative rates for the immunodiffusion test and BAM/AOAC methods were 3.6 and 1.7%, respectively. There was no significant difference in the productivity of the immunodiffusion test and BAM/AOAC method at the 5% level for any of the 6 foods. The immunodiffusion screening method has been approved official first action for detection of motile Salmonella in foods.

A comparison of two types of scalders was conducted to determine their effectiveness in reducing bacterial contamination of poultry carcasses. A conventional hot-water scalder and a prototype model of a steam scalder were tested under commercial conditions. Total plate counts from steam-scalded birds were significantly lower than the counts of water-scalded birds immediately after scalding and again after picking. No differences in the two methods could be found after chilling. Coliform counts from steam-scalded birds were significantly lower than the counts from water-scalded birds immediately after scalding. No significant differences in coliform counts were detected when the two scald methods were compared after defeathering and chilling.

Bacteriological examination of 1,427 samples from a poultry-processing plant over a 2-year period yielded 202 (14.2%) cultures positive for salmonellae. The results indicate that contamination is reduced by washing procedures within the plant but that recontamination of the carcasses occurred in at least two different stages of processing, i.e., during evisceration and chilling. There was evidence of spread of salmonellae from flock to flock during the serial processing of flocks, but the spread was usually not extensive. The serotypes of salmonellae isolated in this study were similar to those of chicken origin reported from other areas of the country.

Studies were conducted to ascertain the bacteriological condition of commercially cooked Eastern-type (foil-wrapped-oven roasted) turkey rolls during processing and storage. After 2 weeks at 5 C, numbers of aerobes on the surface of rolls, in slices, and in whole rolls reached levels of from 1 to 10 million per cm(2) or per g. In stored whole rolls, coliform and enterococcus counts ranged, respectively, from about 10,000 to more than 1 million per g and from < 100 to more than 1 million per g. Postcooking processing operations in two plants did not significantly affect the total count of turkey rolls. Eight of 28 rolls obtained after handling and packaging contained coagulase-positive staphylococci.

Optimum blending fluids and blending times for use in quantifying bacteria on poultry carcass skin by the skin "blending" method were determined. Butterfield's buffered-phosphate diluent, physiological saline solution (0.85% NaCl), peptone water (0.1% peptone), and deionized water, each at four different skin blending times of 1, 2, 3, and 4 min, were compared. The comparison was based on relative numbers of bacteria per cm(2) of skin, enumerated by each combination on turkey carcasses. Peptone water and physiological saline solution each yielded significantly (P < 0.01) higher bacteria counts from turkey carcass skin samples than did Butterfield's buffered-phosphate diluent or deionized water. There were no significant differences among the four skin blending times and no significant interaction effect between the two factors tested.

Bacteriological analyses were made on 45 swab samples secured from hands of poultry workers on processing line, on 31 bruised and 15 normal poultry tissue samples, and on 15 swabs obtained from infected lacerations and exudates of abcesses on hands, arms, chest, and abdomen of poultry workers. A total of 170 Staphylococcus cultures were isolated from samples examined. These cultures were characterized morphologically and biochemically and then grouped into six distinct patterns. S. aureus was found in 86.6% of swab samples obtained from infected workers, in 40% of swabs from hands of workers who handle bruised birds, and in 38.7% of bruised tissues, and was absent from all samples obtained from hands of workers who do not handle bruised birds. All the coagulase-positive staphylococcal isolates were bacteriophage-typed, and the results showed that the same phage-type S. aureus was found in many poultry bruises and in infected lesions of poultry workers as well as on hands of workers who handle bruised birds. These results indicate that poultry bruises are a source of staphylococcal infection encountered among poultry workers.

A comparison was made of four methods of thawing frozen chickens and an average thaw time for each method was determined.Fully and partially thawed chickens, inoculated with salmonellas, Clostridium welchii and Staphylococcus aureus were cooked in a spit-roasting oven at different temperatures for different lengths of time. The chickens were examined freshly cooked and after storage under various conditions.Spit roasting fully thawed chickens until the outer skin was golden brown was sufficient heat-treatment to kill salmonellas and Staph. aureus but Cl. welchii could survive. Salmonellas could also survive if the chickens were not fully thawed before cooking.Incorrect storage after cooking was shown to encourage the growth of pathogens.The incidence of intestinal pathogens in frozen dressed chickens and environmental hazards in spit-roasting establishments were also studied. Of raw chickens examined 35% contained salmonellas (9 serotypes), 63% contained Cl. welchii and 63% Staph. aureus.