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.     

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.     

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.     

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.     

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.     

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     

HYGIENIZATION OF INDIAN CHICKEN MEAT BY IONIZING RADIATION

Both fresh and frozen chicken meat were evaluated for microbiological status by screening for total bacterial counts and for the presence of pathogens like Enterobacteria , Bacillus cereus, coagulase positive Staphylococci and Salmonella spp. Most of the samples exhibited heavy bacterial contamination (1.2 × 10⁵ - 2.6 × 10⁶/g), mainly with Staphylococcus spp. (1.5 × 10⁴ - 2.8 × 10⁵/g). All the chicken samples also showed the presence of Salmonellae (3 × 10¹ - 2.1 × 10²/g). Among the different serotypes observed in chickens . S. typhimurium was common in fresh as well as frozen chicken. Radicidation at 2 kGy at cryogenic conditions (−40°C) was efficient in eliminating the natural pathogenic contamination of the poultry . Salmonella spp. viz. S. seftenberg and S. typhimurium differed in radiation sensitivity, the D₁₀ values in phosphate buffer (pH 7.2) being 0.25 kGy and 0.12 kGy, respectively. Chicken homogenate (10%) offered approximately 2-fold protection to these cells. Chicken samples artificially inoculated with a heavy inoculum (10⁸ cells/g) of these 2 serotypes required higher gamma radiation doses of 4–5 kGy. The findings suggested that a dose of 2 kGy is adequate for normally contaminated chicken samples, but for the heavily contaminated chicken a dose of 4–5 kGy, depending upon the predominating Salmonella serotype present, is required .     

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 .     

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.     

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.     

EFFECT OF SEVERAL EXPERIMENTAL PARAMETERS ON COMBINATION OF RED KIDNEY BEAN (PHASEOLUS VULGARIS) α-AMYLASE INHIBITOR WITH PORCINE PANCREATIC α-AMYLASE

Purified red kidney bean (Phaseolus vulgaris) amylase inhibitor forms a 1:1 stoichiometric complex with porcine pancreatic α-amylase leading to complete loss of enzyme activity on starch. Rate of complex formation is pH dependent and is maximal at pH 5. The rate constants for complex formation, as measured by loss of amylase activity, were 2.85 × 10⁴ M^-1 sec^-1 at pH 6.9 (ionic strength of 0.918) and 2.55 × 10⁵ M^-1 sec^-1 at pH 5 at 30°C. At pH 6.9, rate of complex formation was 4.8 times faster at 0.918 ionic strength as compared with the rate at 0.138 ionic strength. At 30°C, pH 6.9 and ionic strength of 0.168 the dissociation constant of the enzyme-inhibitor complex was determined to be 3.5 × 10^-11 M. The rate constant for dissociation of the complex was calculated to be 8.7 × 10^-8 sec^-1 under the same conditions. The rate constant for complex formation, at ionic strength of 0.168, was 1.1 × 10⁴ M^-1 sec^-1 at 37⁰ and 9.77 × 10² M^-1 sec^-1 at 25.7°C. The calculated activation energy for complex formation is 39.5 kcal/mole suggesting a rate-controlling conformational change. Oxidation of the carbohydrate moiety of the glycoprotein inhibitor caused complete loss of activity. Maltose, a competitive inhibitor of α-amylase, bound as readily to the enzyme-inhibitor complex as to free α-amylase. Trypsinized α-amylase, although still able to bind to Sephadex, did not bind inhibitor. The experiments with maltose and trypsinized amylase suggest the inhibitor may not bind at the active site of α-amylase.     

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.     

Singlet Oxygen Oxidation Rates of ∝-, γ-, and δ-Tocopherols

ABSTRACT:  The reaction rate constants of 5 × 10⁻⁴ M, 10 × 10⁻⁴ M, and 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols with singlet oxygen in methylene chloride containing 1 × 10⁻⁵ M chlorophyll under light at 25 °C for 60 min were studied. The oxidation of tocopherols determined by a spectrophotometric method showed that the losses of 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols after 60 min under light were 21%, 16%, and 9%, respectively. The degradation of α-, γ-, and δ-tocopherols was undetectable in the absence of chlorophyll under light or in the presence of chlorophyll in dark. The losses of tocopherols under light were mainly due to singlet oxygen oxidation. The degradation rates of 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols were 6.6 ×10⁻⁶ M/min, 5.0 × 10⁻⁶ M/min, and 2.9 × 10⁻⁶ M/min, respectively. The reaction rates between α-, γ-, or δ-tocopherol and singlet oxygen were 4.1 ×10⁶/M s, 3.3 × 10⁶/M s, and 1.4 × 10⁶/M s, respectively. The singlet oxygen oxidation rate of δ-tocopherol was significantly lower than α- or γ-tocopherol at α= 0.05. As the electron density in the chromanol ring of tocopherol increased, the singlet oxygen oxidation was increased.     

Riboflavin Photosensitized Singlet Oxygen Oxidation of Vitamin D

Samples containing various levels of vitamin D and riboflavin, with and without ascorbic acid or a-tocopherol were prepared in a model system. Samples were stored in the light or in the dark at 45®C for up to 8 h. Headspace oxygen was determined by gas chromatography with thermal conductivity detection. Oxidation of vitamin D was not observed in samples without riboflavin stored in the light nor in samples with riboflavin stored in the dark. Vitamin D with riboflavin was oxidized under light. α-Tocopherol quenched singlet oxygen at a rate of 2.52 × 10⁸ M^-1 sec^-1, whereas ascorbic acid quenched singlet oxygen at a rate of 2.23 × 10⁷ M^-1 sec^-1.     

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.     

Production of Enterotoxin-B in Cured Meats

SUMMARY— A variety of laboratory cured hams were inoculated with 10³−10⁶ cells of S. aureus strain S-6 and incubated at 10, 22 and 30°C anaerobically for up to 16 weeks. Enterotoxin-B was detected by gel-diffusion in hams with original pH over 5.30, up to 9.2% NaCl (brine) and 0.54 ppm undissociated nitrous acid. There was better toxin production at 30° than at 22° or 10°C. Toxin was detected at 10°C after at least 2 weeks incubation and in most samples after 8 weeks when pH was greater than 5.6. Toxic hams had more than 4 × 10⁶ cells/g. Contaminants were always less than 10⁵/g. Tween 80 inhibited toxin production at 30° but not at 10°C. Toxic hams looked normal even after 2 months incubation at 10°C.     

SPOILAGE AND HISTAMINE IN WHOLE PACIFIC HERRING (CLUPEA HARENGUS PALLASI) AND PINK SALMON (ONCORHYNCHUS GORBUSCHA) FILLETS

Fresh pink salmon (Oncorhynchus gorbuscha) fillets and whole Pacific herring (Clupea harengus pallasi) were stored for two weeks at 10C to determine if significant amounts of histamine were produced before the fish spoiled. Spoilage odors in salmon were moderate by day 4 and intense by day 7, while herring had detectable spoilage by day 4 and became potent by day 6. Aerobic bacterial counts increased from 4.0 × 10²/g initially to 3.6 × 10⁸/g in salmon fillets by day 14 and from 2.3 × 10³/g initially to 2.7 × 10⁷/g in whole herring by day 14. Total volatile nitrogen increased from 1.8 to 78.5 mg N/100 g in salmon and 2.2 to 23.6 mg N/100 g in herring. Histamine was not detected in salmon, while concentrations reached 54.9 ppm in herring at day 14. However, herring were considered spoiled by day 6.     

An Objective Evaluation of Changes in Firmness of Ripening Bananas Using a Sonic Technique

SUMMARY— Firmness in fruits is sometimes evaluated as being the force necessary to attain a given deformation within the product. Since modulus of elasticity is defined as the ratio of stress to strain, it should measure resistance to force and, hence, the firmness of a material. Using a sonic technique, the resonant frequencies of cylindrical specimens of flesh from Valery bananas were measured and Young's modulus of elasticity was calculated. Softening of the banana during ripening was associated with a decrease in Young's modulus of elasticity from 272 × 10⁵ dynes/cm² at ripeness corresponding to a light green peel color to 85 × 10⁵ dynes/cm² at the yellow stage. Modulus of elasticity was significantly and directly correlated with starch content, but inversely correlated with luminous reflectance and the logarithm of percent reducing sugars.     

Influence of pre-treatments on microbial load of stored dehydrated onion slices

Onions slices were pre-treated in potassium meta-bisulphite (KMS) and sodium chloride with different concentration levels to study the microbial load of tray and greenhouse-dried onion slices up to 6 months of storage. Data were analysed as per procedure of one-way classified anova using DMRT of AgRes statistical package for bacteria, yeast, fungi and Lactobacilli . Results revealed that in almost all samples, permissible levels of bacteria [18.33 × 10¹ colony-forming units (CFU) per gram], yeast (ND), fungi (0.5 × 10¹ CFU g⁻¹) and Lactobacilli (0.25 × 10¹ CFU g⁻¹) were observed after 5 months of storage. All the samples were also found to be free from Escherichia coli and no Mac Conkey growth was noticed. Onion slices pre-treated in 0.25% and 0.50% KMS and dried in tray and greenhouse, respectively, were found best after 6 months of storage period.     

Microbiological quality of ice creams commercialized in some cities in the state of Rio de Janeiro, Brazil

The microbiological quality of 60 ice cream samples of three commercial brands (A, B and C) of various flavours, commercialized in some towns in the State of Rio de Janeiro, Brazil, was evaluated. Total bacteria count (TCB), coliforms at 35°C (CT), coliforms at 44°C (CF) and presence of Staphylococcus aureus were performed on all samples. TCB ranged from 2.0 × 10 ² to 6.9 × 10 ⁵ cfu/mL, CT from < 3–≥ 2400 MPN/mL, CF from < 3–1100 MPN/mL and S. aureus from < 10–1.4 × 10 ⁶ cfu/mL. The level of bacterial contamination found in this study reflects the unhygienic conditions prevalent in manufacturing and storage of ice creams. Actions are thus necessary by the Brazilian regulatory agencies to require the ice cream processing plants to adopt quality guarantee systems, such as good manufacturing practices and hazard analysis and critical control points system.