REMOVAL OF SALMONELLA TYPHIMURIUM ATTACHED TO CHICKEN SKIN BY RINSING WITH TRISODIUM PHOSPHATE SOLUTION: SCANNING ELECTRON MICROSCOPIC EXAMINATION

The effect of trisodium phosphate (TSP) on Salmonella typhimurium attached to chicken skin was investigated by using scanning electron microscopy (SEM). Chicken drumsticks were inoculated with Salmonella typhimurium (2 × 10⁸ CFU/mL) for 30 min. Both inoculated and non-inoculated drumsticks were rinsed with 10% TSP solution at 10 or 50C for 15 s, and skin pieces were cut and fixed for SEM examination. For inoculated skins, a significant difference was noticed between TSP-rinsed and control skins (water-rinsed) at both temperatures. While control skins were covered with salmonellae (4 × 10⁵∼ 1 × 10⁶ CFU/cm²) and miscellaneous debris, TSP-rinsed skins, either at 10 or 50C, showed clean skin surfaces (<8×10³ CFU/cm²). For non-inoculated skins, it was difficult to see the difference in the number of attached bacteria due to their low numbers, however, water-rinsed skins still showed the debris on the surface. Above observations suggest that one of the major mechanisms of TSP on salmonellae reduction is detachment of contaminants from the skin surface.     

MICROBIOLOGICAL QUALITY OF THE DAIRY PRODUCT PEDHA AND ITS IMPROVEMENT USING GAMMA RADIATION

Eighteen pedha samples procured from A and B grade retail shops were examined for their overall microbiological quality and for the presence of foodborne pathogens viz. Staphylococcus aureus, Salmonella sp., Coliforms , Listeria monocytogenes, Yersinia enterocolitica and Bacillus cereus. The microbiological quality of pedha samples from B grade shops was very poor as compared to pedha from A grade shop as evidenced by the very high total bacterial counts (6 × 10⁷ cfu/g), high counts of S. aureus (as high as 7 × 10⁶ cfu/g) and presence of coliforms and Listeria and Yersinia sp. in 33% of the samples. All the samples from A grade shops were also positive for S. aureus though negative for coliforms , Yersinia, Salmonella, Listeria and B. cereus. Gamma irradiation of pedha at a dose of 3kGy at 0C reduced overall bacterial load by five log cycles and S. aureus and coliforms could be totally eliminated. However, 5 kGy dose was necessary to eliminate S. aureus if the initial number exceed 1 × 10⁵ cfu/g. Inoculated pack studies confirmed that 3 kGy dose was sufficient for the complete elimination of up to 1 × 10⁵ cfu/g of S. aureus. A dose of 3 kGy had minimal effect on the sensory quality of pedha and even pedha samples irradiated at 5 kGy dose were acceptable. Treatment with 3 kGy dose of gamma radiation totally eliminated S. aureus and coliforms in pedha, thereby ensuring their microbial safety.     

MICROBIOLOGICAL STATUS OF EGYPTIAN SALTED MEAT (BASTERMA) AND FRESH SAUSAGE

The microbiological quality of 125 samples of the most popular Egyptian meat products (75 Egyptian fresh sausage "EPS" and 50 basterma) was determined. The aerobic plate count (APC) and Lactobacillaceae count of basterma ranged from 1 × 10⁴ to 9 × 10⁶ CFU/g, respectively . Enterobacteriaceae, mold and yeast counts for basterma were similar (<1 × 10² CFU/g). Artificially contaminated slices of meat and garlic paste of basterma showed that the paste inhibited growth of Salmonella typhimurium. APC and Enterobacteriaceae counts of EFS ranged from 1.1 × 10⁴ to 1 × 10⁸ and from 1 × 10² to 1 × 10⁷ CFU/g, respectively. Nearly 26% and 29% of EFS were positive for Clostridium perfringens and coagulase-positive Staphylococcus aureus, respectively . Salmonella could not be detected in any examined samples. Bacteriologically, EFS might pose a potential health hazard, making it imperative to institute sanitary measures during its production and sale .; Accepted for Publication July 2, 1997     

SHELF-LIFE OF FRESH FILLED PASTA. HAZARD ANALYSIS AND CRITICAL CONTROL POINTS OF THE MANUFACTURING PROCESS AND HOUSEHOLD PRACTICES

Hazard Analysis and Critical Control Point (HACCP) approach was used to assess the safety of a fresh filling pasta (ricotta-filled ravioli) manufacturing plant in La Plata region, Argentina. Household practices like cooking and holding meals before serving were also evaluated. Samples of ricotta, main raw material of the filling, showed colony counts of Enterobacteriaceae total microorganisms, molds and yeasts of (5 × 10³-1 × 10⁵), (1x10⁵−1x10⁸) and 1 × 10⁵ CFU/g, respectively. E. coli was also detected in one out of five samples, suggesting a hazardous condition of this raw material. Salmonella spp. was not isolated from any of the dough samples tested. Ricotta filling showed high colony counts of mesophilic microorganisms (6 × 10⁶ CFU/g), Enterobacteriaceae (1 × 10⁵ - 1 × 10⁶ CFU/g), while E. Coli was detected in 20% of the samples. Counts of B. cereus, S. aureus were less than 1 × 10² CFU/g in the analyzed materials. In the finished product (ricotta-filled ravioli), the colony counts of mesophilic microorganisms and Enterobacteriaceae were 3 × 10⁸ and 8 × 10⁵ CFU/g, respectively. Critical Control Points found were cooking and holding time before serving. The implementation of suggested corrections allowed the microbial quality of the final product (ricotta-filled ravioli) to be improved considerably. The growth of total microbial counts during refrigerated storage (0, 4, 8 and 10C) were measured and the shelf-life of ricotta and ricotta-filled ravioli was calculated.     

EFFECT OF ELECTRICAL STIMULATION ON KILLING SALMONELLA TYPHIMURIUM IN VARIOUS SALT SOLUTIONS

Electrical stimulation was evaluated as a method to kill Salmonella typhimurium in various salt solutions at different concentration . Salmonella typhimurium at 2 × 10⁵ CFU/ml was treated at 22–24C for 60 min in each salt solution using electricity at 10 mA/cm² current, 1 kHz frequency, and 50% duty cycle. Samples taken at various times were serially diluted, plated on tryptic soy agar and xylose lysine desoxycholate agar, and incubated at 37C for 18–24 h. To detect injured cells, samples were also pre-enriched in buffered peptone water at 37C for 4–5h before being plated. Results indicated all salmonellae were electrically killed at 5 min in NaCl, at 30 min in NaNO₃, and at 45 min in NaC₂H₃O₂ at 0.15 and 0.015 M concentrations. Salmonellae were also killed at 45 min in Na₃PO₄ and at 60 min in Na₂CO₃ at 0.0015 M concentration by electricity in combination with high pH .     

QUANTITATIVE STUDIES OF PROTEOLYTIC AND LIPOLYTIC BACTERIA IN FROZEN MINCED BEEF AND HAMBURGER

Sixty frozen samples (30 hamburger patties and 30 minced meat) purchased from different retail markets in Ismailia, Egypt were examined for incidence of proteolytic and lipolytic bacteria. Mean values of proteolytic psychrophiles in the examined samples of hamburger and minced meat were 2×10⁵ and 6×10⁴, respectively. Lipolytic psychrophiles in hamburger were 8×10⁵ and 3×10⁵ in minced meat. Proteolytic mesophiles in hamburger and minced meat were 6×10⁵ and 5×10⁴, respectively. Mean values of lipolytic mesophiles were 5×10⁴ for hamburger and 5×10³ for minced meat. Proteolytic thermophiles in hamburger and minced meat were 3×10³ and 10³, respectively. Both proteolytic and lipolytic activities were exhibited by various bacteria that were identified.     

ELIMINATION OF SALMONELLA FROM FROZEN SHRIMP BY GAMMA RADIATION

Shrimp samples were found to be contaminated with Salmonella. The D₁₀ values of S. typhimurium and S. enteritidis were between 0.225 to 0.250 kGy when cells were irradiated in frozen 0.1 M phosphate buffer. The radiation resistance of these Salmonella spp. increased marginally when cells were irradiated in shrimp homogenate. A radiation dose of 4.0 kGy completely eliminate Salmonella from frozen prepackaged shrimp, which was confirmed by inoculated pack studies .     

SURVIVAL OF THREE SALMONELLA SEROTYPES ON BEEF TRIMMINGS DURING SIMULATED COMMERCIAL FREEZING AND FROZEN STORAGE

This study investigated the survival of three Salmonella serotypes (S. Brandenberg, S. Dublin and S. Typhimurium) on beef trimmings during simulated commercial freezing, frozen storage for 9 months and subsequent abusive slow thawing and refreezing conditions. This was achieved by plating samples monthly and after thawing and refreezing on nonselective Tryptic Soy Agar (TSA) and selective Xylose Lysine Desoxycholate Agar (XLD) and incubating both at 37C for 24 h to determine Salmonella counts, aerobic counts and the presence, if any, of sublethal injury of this pathogen. Two freezing temperatures (−18C or −35C) to simulate slow or rapid freezing respectively, and two inoculation levels (10³ cfu g⁻¹ or 10⁵ cfu g⁻¹) were used. Aerobic counts and counts of all the Salmonella serotypes did not change significantly (p > 0.05) during frozen storage or for any of the other treatments applied in this study. This finding was attributed to the insulating nature of the subcutaneous fat layer in this manufacturing cut. These results are important with respect to food safety associated with ground beef processing.     

DETECTION OF SALMONELLAE IN CHICKEN AND FISH SAMPLES USING DNA PROBE

The use of DNA probes for rapid identification of foodborne pathogens is an emerging field. We have standardized a method for the detection of Salmonella in food samples by using a specific insertion sequence (IS-200) of Salmonella chromosomal DNA as the probe. The procedure for colony hybridization, and washing conditions for the stringency were established. All the ten Salmonella strains, belonging to various serotypes tested were positive while 18 other bacterial cultures, mainly belonging to the family Enterobacteriaceae such as E. coli Shigella, Proteus etc., were negative in colony hybridization. The sensitivity of the detection was found to be 10⁵ cfu using ³²P-labeled probe. We surveyed 16 chicken and 6 fish samples from local market using IS-200 probe after preenrichment and selective enrichment. All the samples tested were found positive with DNA probe. The major strains of Salmonella isolated from these samples were S. gallinarum, S. typhimurium, S. enteritidis, S. choleraesuis and S. paratyphi A. These results were confirmed by standard Salmonella identification method.     

MICROBIOLOGICAL QUALITY OF TRADITIONAL MARKET CURED FISH IN TANZANIA

The microbiological quality of six varieties of retail market traditionally cured fish in Morogoro, Tanzania was investigated over a five-month period. The fish were contaminated with bacteria and molds at levels of: total aerobes, 10⁶ - 1.7 × 10⁷ c.f.u/g; faecal coliforms, 1.1 × 10¹ - 2.5 × 10³ MPN/g; faecal streptococci, 1.4 × 10¹ - 1.3 × 10³ MPN/g; Staphylococcus aureus, 1.3 × 10³ - 8.6 × 10³ c.f.u/g; Aspergillus flavus group, 2.1 × 10¹ - 2 × 10² c.f.u/g of fish. Of faecal coliform, 45% of the isolates were Escherichia coli. Twenty-five percent of the S. aureus isolates were coagulase positive. Sixteen percent of A. flavus isolates were aflatoxigenic. Aflatoxin contamination ranged from O to 18.5 μg/kg of fish. Insect infestation by Dermestes spp. and mites was observed. The results of this preliminary study emphasize the importance of proper processing and handling offish in the tropics in order to safeguard public health .     

RADIATION STERILIZATION OF SPICES FOR HOSPITAL FOOD SERVICES AND PATIENT CARE

A survey of the commercial spices used by food services in a typical hospital environment revealed high contamination with microorganisms, i.e., 10⁴ to 10⁷ counts per gram. The predominant microorganisms were as followed (in colony counts/gram): (1) heat-resistant bacterial spores in black pepper, 1 × 10⁷; thyme, 2 × 10⁶; anise, 7 × 10⁴; curry powder, 4 × 10⁵; poultry seasoning, 8 × 10⁴; pickling spice, cardamom, and cumin, 1.5–3 × 10⁴; (2) mixed populations of vegetative cells and bacterial spores in cumin, 1 × 10⁶; (3) molds in cream of tartar, 2 × 10⁴. Sterility of food may be important in a hospital setting, especially in the care of immunocompromised patients. To eliminate the organisms, we recommend radiation treatment, accompanied by appropriate microbiological quality control. On the basis of radiation survival data, the composite natural flora would be reduced to the level of "commercial sterility" (defined as less than 10 organisms per gram((Kiss 1982) by the following minimum radiation doses (in kGy): black pepper, 13; thyme, 13; cumin, 12; anise, 10; curry, 7.3; pickling spice, 7; poultry seasoning, 6; cardamom, 9.4; cream of tartar, 4. For practical purposes, two dose levels can be recommended for treatment of spices in the hospital environment, low = 6–10 kGy and high = 10–15 kGy.     

EXAMINATION OF THE MICROBIAL STATUS OF SELECTED INDIGENOUS FERMENTED FOODS IN NIGERIA

Four Nigerian traditionally fermented foods (wara, nono, ogi and kununzaki) were evaluated for the presence of some microorganisms of public health concern. Among the dairy foods , Staphylococcus aureus and Klebsiella sp. were isolated from wara while Escherichia coli, Salmonella sp. and Klebsiella sp. were isolated from nono. The cereal-based fermented foods (ogi and kunu-zaki) contained Bacillus subtilis, E. coli, S. aureus, Klebsiella sp. and Enterococcus faecalis. The mesophilic aerobic counts were: 5 × 10⁵ for wara; nono, 1.53 × 10⁷; ogi, 3.6 × 10⁶ and kunu-zaki, 2.6 × 10⁶ cfu/mL. The enterobacteriaceae counts on nono, wara, ogi and kunu-zaki were 1.79 × 10⁷, 4.5 × 10⁵, 4.0 × 10⁵ and 1.2 × 10⁶ cfu/mL, respectively. No Vibrio count (detection limit: <10 cfu/mL) was recorded in all the food samples considered. The yeast and mold counts ranged from 1.0 × 10⁵– 3.31 × 10⁷ among the food products. The antimicrobial susceptibility patterns of the organisms isolated from dairy products (nono and wara) revealed that they were resistant to ampicillin (100%) and sensitive to gentamicin (100%) and nalidixic acid (100%). Most isolates from cereal based products (ogi and kunu-zaki) were 100% resistant to penicillin, ampicillin and chloramphenicol. This work highlights the need to maintain hygienic standards in the preparation of our locally fermented cereal and dairy foods.     

Effect of irradiation and modified atmosphere packaging on the microbiological safety of minced pork stored under temperature abuse conditions

The safety of irradiated pork packed in 25% CO₂:75% N₂ and stored at abuse temperature (10 or 15°C) was assessed by inoculation studies involving Salmonella typhimurium, Listeria monocytogenes, Escherichia coli, Yersinia enterocolitica and Clostridium perfringens . Irradiation to a dose of 1.75 kGy reduced pathogen numbers to below the detection limit of 10² cells g^-1. When higher inoculum levels were used (10⁶ cells g^-1) irradiation at 1.75 kGy reduced pathogen numbers by 1 –>5 log₁₀ cycles depending on strain. Clostridium perfringens was the most resistant, and Y. enterocolitica the most sensitive of the pathogens studied.
In all cases when high numbers (10⁶ to 10⁷g^-1) of spoilage and/or pathogenic bacteria were present initially on the pork the meat appeared spoiled, and although irradiation reduced the number of microorganisms, the meat was still unacceptable from a sensory viewpoint after treatment.
It was concluded that the microbiological safety of irradiated, modified atmosphere packaged (MAP) pork is better than that of unirradiated MAP pork.     

Piezoelectric Flow Injection Analysis Biosensor for the Detection of Salmonella Typhimurium

ABSTRACT: Piezoelectric biosensors have the potential to provide direct detection of food contaminants, such as pathogens. In this study, Protein A antibody immobilization was used for the activation of the piezoelectric biosensor to detect Salmonella typhimurium. The overall system consisted of a new design for a flow cell and flow injection analysis system. The flow cell made possible a baseline stability of ± 1 Hz out of 5 MHz for hours. The sensor had responses of 5 to 65 Hz in 30 min with R²= 0.95 for S. typhimurium concentrations of 10⁷to 10⁹ CFU/ml under continuous flow, and 3 to 75 Hz in 40 min with R²= 0.96 for S. typhimurium concentrations of 10⁶ to 10¹⁰ CFU/ ml under stop flow. Cross-reactivity tests were performed with nonpathogenic Escherichia coli, Escherichia coli O157:H7, Listeria monocytogenes , and Vibrio parahaemolyticus and showed less than 10% response.     

EFFECT OF HIGH HYDROSTATIC PRESSURE, GAMMA-IRRADIATION AND COMBINATION TREATMENTS ON THE MICROBIOLOGICAL QUALITY OF LAMB MEAT DURING CHILLED STORAGE

The effect of gamma irradiation (1.0 kGy) and high hydrostatic pressure (200 MPa for 30 min), either alone or in combination on the shelf-life of lamb mince meat at 0–3C was studied. Untreated control samples initially had total microbial counts of 10⁵ CFU/g, 10² CFU/g of coliforms and 10⁴ CFU/g of Staphylococcus spp. Coliforms were eliminated by all the treatments . Staphylococcus spp. however, were reduced only by 1 log cycle when treated with irradiation alone and high pressure alone. These species were a mixture of mannitol-fermenting and mannitol-nonfermenting strains. In samples subjected to the combination treatment , Staphylococcus spp. appeared only after 3 weeks of storage and all were mannitol-nonfermenting. On the basis of microbiological and sensory quality, the shelf-life of the control sample was less than 1 week. All treated meat samples had a shelf-life of 3 weeks, but only combination treated samples were free from potentially pathogenic Staphylococcus spp .     

INHIBITION OF STAPHYLOCOCCUS AUREUS AND BACILLUS CEREUS ON A VEGETARIAN FOOD TREATED WITH NISIN COMBINED WITH EITHER POTASSIUM SORBATE OR SODIUM BENZOATE

Various amounts of nisin (0, 10³ and 5 × 10³ IU/g) in combination with either potassium sorbate (0, 2, and 3%) or sodium benzoate (0, 0.06 and 0.12%) were tested for effectiveness in inhibiting growth of Staphylococcus aureus C10 and Bacillus cereus B7 inoculated on a vegetarian food. The strains used were isolated from vegetarian foods obtained commercially in Taiwan, and the test food, spice and dried bean curd, was selected for the study based on ability to support the growth of these organisms. After treatment with a preservative combination, the surfaces of sterilized food samples were inoculated, samples were stored in vacuum or nonvacuum packages at either 4C or 30C, and at appropriate times, tested for microbial growth. Growth of both isolates was unaffected by vacuum-packaging treatment; however, a bacteriostatic effect was found at 4C. Data indicated that during the 14-day storage at 4C, vacuum-packaged samples treated with 5 × 10³ IU/g nisin and 0.12% sodium benzoate significantly (p < 0.05) decreased the counts of S. aureus C10 and B. cereus B7 by 2.61 and 3.02 log₁₀ CFU/g, respectively. In the vacuum-packaged samples treated with 5 × 10³ IU/g nisin and 3% potassium sorbate, counts for C10 and B7 were decreased by 2.35 and 2.64 log₁₀ CFU/g, respectively. Thus, the combined treatment extended the shelf-life of the vegetarian food .     

LINKING MICROBIOLOGICAL CRITERIA FOR FOODS WITH QUANTITATIVE RISK ASSESSMENT

The current annual risk of acquiring a foodborne disease in the United States is estimated at 2.7 × 10^-2. The risk of associated death is estimated at 3.7 × 10^-5. These represent a health care burden >$3 billion. Using a risk assessment model one can identify levels of microbial contamination which may be unacceptable in foods and appropriate controls needed to reduce these levels. Salmonella bacteria continue to represent a large percentage of the identifiable infections. A model developed from human dose-response studies predicts the probability of infection for Salmonella at 7.5 × 10^-3 with exposure to a single CFU of the organism. Risks of severity (hospitalization), mortality, reactive arthritides, and mortality in the elderly are estimated at 3.1 × 10^-6, 7.5 × 10^-6, 1.7 × 10^-5, and 2.8 × 10^-4. Exposure to microbial contaminants needs to be evaluated on a single meal basis. For chicken, exposure may range from a single drum stick (38g) to a half broiler (176g) but averages around 80g. For beef between 51 and 85g may be consumed during a single meal. Therefore, methods for monitoring must be able to detect at least 1 CFU/80g. Risks for some pathogenic E. coli are estimated at 1,000 to 10,000 less than Salmonella. Therefore, use of coliforms as indicators needs to be assessed and related to occurrence and survival and regrowth potential of the enteric bacteria of greater public health concern. Because, 20% of the U.S. population may be considered to be in a special population category and at an increased risk of severe outcomes, no more than 20% failure of a standard should be acceptable .     

LACTIC ACID INACTIVATION OF SALMONELLA TYPHIMURIUM ATTACHED TO CATFISH SKIN

This study was conducted to test sanitizer efficacy of lactic acid on Salmonella typhimurium populations that were firmly attached or loosely attached on catfish skin with or without mucus. For both cell types, counts of S. typhimurium were reduced from 5.9 log¹⁰ CFU/skin to below the limit of detection of 2.0 log¹⁰ CFU/skin by a 5 min exposure to 2% lactic acid. Exposure for 1 min reduced a 10⁴ CFU/skin inoculum to below the limit of detection. Catfish skin mucus slightly decreased the antimicrobial effect of lactic acid against firmly attached S. typhimurium.     

RHEOLOGICAL AND CHEMICAL PROPERTIES OF MOZZARELLA CHEESE

Dynamic viscoelastic parameters and chemical properties of Mozzarella cheese produced using a "no-brine" cheese making method with 3 different cooking temperatures (38, 41, and 44C) were determined. Samples were stored for 3 weeks at 4C before dynamic mechanical analysis at 22C. G', G" and tan δ were 5.8 – 6.4 × 10⁵ dyne/cm², 1.9 – 2.1 × 10⁵ dyne/cm², and 0.33 – 0.35, respectively, at 1% strain and 10 rad/s. The percentage of intact α_s-casein and β-casein were 38–40% and 33–35% of total protein in the cheese, respectively. The range of cooking temperatures used in this experiment had little effect on dynamic viscoelastic properties or the amount of intact protein for the cheese.     

Rapid dye reduction tests for the determination of microbiological quality of meat

Rapid dye reduction tests have been developed to determine the quality of meat. Three chemical indicators, resazurin and two tetrazolium compounds, were used to correlate the microbial numbers and reduction times in meat samples. Twenty-five surface samples from sheep carcasses were subjected to each reduction test. Total viable counts given were obtained at 37°C. Resazurin reduction time was 90–120 min when the bacterial counts ranged from 1.5×10⁶ to 7.7×10⁶/cm². Samples showing bacterial counts between 1.5×10⁶ and 6.0×10⁶/cm² reduced tetrazolium (NBT) in 360–390 min whereas samples containing bacterial counts of 2.1×10⁶/cm² took 420–450 min to reduce iodophenyl nitrophenyl tetrazolium (INT) dye. Regression equations relating the number of organisms per cm² and reduction time were applied to predict the microbiological quality of meat samples from reduction time data. Among the three dyes, resazurin gave the lowest reduction time.