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.     

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.     

RELEASE OF PROPYL PARABEN FROM A POLYMER COATING INTO WATER AND FOOD SIMULATING SOLVENTS FOR ANTIMICROBIAL PACKAGING APPLICATIONS

The release phenomena of propyl paraben from a polymer coating to water and three food simulating solvents (10% aqueous ethanol, 50% aqueous ethanol, n-heptane) were studied for antimicrobial packaging applications. The effects of food simulating solvent, initial concentration in the coating and temperature on the propyl paraben release were examined. The initial concentration of propyl paraben in the coating ranged from 1.26 × 10⁴ to 10.52 × 10⁴ g/m³ and the temperature from 5.5 to 30C. For water, the release was controlled by Fickian diffusion with constant diffusion coefficient (7±11 × 10^-11 cm²/s at 30C), and independent of the initial concentration. For 10% ethanol, the release followed again the Fickian model with constant diffusion coefficient (30±40 × 10^-11 cm²/s at 30C). For 50% ethanol and n-heptane, the release was instantaneous and not controlled by Fickian diffusion. For the release into water, the activation energy for diffusion from the Arrhenius relationship was around 88 kJ/mole.     

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.     

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.     

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.     

CHANGES IN SIZE OF GAS CELLS IN DOUGH AND BREAD DURING BREADMAKING AND CALCULATION OF CRITICAL SIZE OF GAS CELLS THAT EXPAND

Microscopic observation showed that a group of small air cells entrained during the early stage of mixing is the original cause of cell structure of bread. At the beginning of fermentation, about 3 × 10⁸/m² gas cells with diameters between 3 × 10⁻⁶ and 8 × 10⁻⁴ m were entrained in the dough. The distribution curve of cell size was approximately normal on a logarithmic scale. During fermentation and proofing, a great portion of carbon dioxide was released into cells larger than about 10⁻⁴ m in diameter that was equivalent to a few percentages of total number of gas cells. After baking, gas cells smaller than 10⁻⁴ m in diameter were not observed and the total number of cells in baked bread reduced to about 10⁶/m² with diameters between 10⁻⁴ and about 5 × 10⁻³ m. The critical cell size to expand generally agreed with the calculated value using an equation, rc'= 3s/E (re': critical radius to expand, s: surface tension, E: elasticity), and cited value of s and E.     

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.     

SIMULATING DIFFUSION MODEL AND DETERMINING DIFFUSIVITY OF POTASSIUM SORBATE THROUGH PLASTICS TO DEVELOP ANTIMICROBIAL PACKAGING FILMS

A diffusion model was simulated by computer programming and diffusivities of potassium sorbate through various plastic films were determined by a lag time method. The simulation showed that partition coefficient affected the flux of total penetration but did not affect the lag time. Therefore, the lag time method is appropriate in determining diffusivity, using any value for the partition coefficient. The diffusivities of potassium sorbate were 1.83 × 10⁻⁸ cm²/s, 4.26 × 10⁻¹³ cm²/s, 4.65 × 10⁻¹³ cm²/s, and 5.47 × 10⁻¹³ cm²/s through low density polyethylene (LDPE), high density polyethylene, polypropylene, and polyethylene terephthalate, respectively at 25 C. Arrhenius equation shows very good fit between the diffusivity and the temperature by the linear regression analysis. D₀ and E_a of potassium sorbate through LDPE film resulted in 1.98 × 10⁻⁶ cm²/s, and 11.83 KJ/mole K. The concentration of potassium sorbate was insignificant and did not contribute to the statistical model. This result verifies that the diffusion of potassium sorbate in LDPE film is typical case ofFickian diffusion.     

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 .     

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 .     

TEXTURE PROPERTIES OF GELLAN GELS AS AFFECTED BY TEMPERATURE

Gellan gels can be made very brittle, similar to agar gels, or very flexible, like gelatin gels. The entropy or enthalpy nature governing those gellan gel behaviors was studied by mechanical testing at temperatures varying from 2 to 62C. Both failure stress and strain for 1% low acyl and low acyl/high acyl mixed gellan gels decreased with increasing temperature, indicating that the hydrogen bonding contributed significantly to the stabilization of gellan gels in addition to the polyanion-calcium-polyanion bonding. Hydrophobic interactions were less important. The initial Young's modulus for two mixed high and low acyl gellan gels containing 2 mM Ca⁺⁺ increased with temperature from 2–42C, indicating entropy elasticity. Average molecular weight between adjacent crosslinks for these two mixed gels was larger than 10⁴. For other gels, the entropy elasticity was not a dominant mechanism for elastic force because of molecular weights between crosslinks and from the observation of negative temperature dependence of the modulus.     

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.     

Texture and mechanical properties of prok backfat

A puncture test for measuring the firmness of backfat is described for use in studies of the effects of changes in animal production, and meat processing on the quality of fatty tissue. The operating and recording conditions were chosen from sensory deformation characteristics used in firmness evaluation when pressing fat with the index finger. On average, assessors exerted aaaximum force of about 30 N and caused a deformation of 4 mm in 2 sec and the best predictor of firmness was the maximum deformation they applied. Mechanical properties of the fats were determined by stress relaxation and the modulus 2.5 sec after compression was about 80 × 10⁴ for hard and < 3 × 10⁴ N/m² for soft fat at 3°C and the logarithm of the modulus was linearly related to the subjective firmness rating.
In the puncture test, the forces required to drive a stainless steel punch (3.5 mm in diameter) 2.5 or 4.0 mm into backfat at 3°C at 2 mm/sec correlated well (r > 0.8) with firmness rating and five replicate determinations using the punch proved more discriminating than a panel of four assessors. A combination of the forces required on inner and outer layers of fat accounted for 93% of variation in firmness assessments.     

Effect of simultaneous high intensity ultraviolet irradiation and hydrogen peroxide on bacterial spores

High intensity (1.8 × 10³ mW/cm²) ultraviolet light irradiation of spores of Bacillus subtilis in the presence of 2.5% (w/v) hydrogen peroxide resulted in a rapid kill. Higher concentrations of hydrogen peroxide protected spores against the effects of irradiation. The results are discussed in relation to the use of simultaneous ultraviolet irradiation and hydrogen peroxide treatment as a method of sterilization.     

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     

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.     

The relationship of bacterial numbers and types to diamine concentration in fresh and aerobically stored beef, pork and lamb

Bacterial numbers and putrescine and cadaverine concentrations were measured in intact beef, pork and lamb and minced beef at retail and during aerobic chill storage at 5°C. Putrescine concentrations increased consistently with 'total' aerobic viable count (TAVC) but cadaverine concentrations increased only when high numbers of presumptive Enterobacteriaceae were present. Significant changes in diamine concentration did not occur until the TAVC exceeded 4.2 × 10⁷/cm² or g when the meat was clearly spoiled. Changes prior to the onset of spoilage were not sufficient for their use as a predictive indicator of the acceptability of the meat.     

Ionizing Radiation Effects on Starch as Shown by Staudinger Index and Differential Thermal Analysis

SUMMARY— Corn starch in the form of raw granules at commercial moisture was irradiated at two levels: 3 × 10⁵ and 6 × 10⁶ rad from a Co⁶⁰ source. The irradiated samples were completely dissolved in alkali, indicating there was no cross linking induced in the starch molecules by irradiation. Viscosity determinations of starch solution diluted with distilled water exhibited the ion charge effect generally observed in other macromolecules.
The Staudinger indices of unirradiated, irradiated at 3 × 10⁶ rad and 6 × 10⁶ rad were 42, 22 and 16 respectively, which were an indication of depolymerization of starch macromolecules with increasing irradiation. The differential thermal analysis of the three samples also showed the depolymerization of the polymer with irradiation.
It is suggested that these two simple techniques–the Staudinger index and D.T.A.–could be usefully employed to characterize small differences in starches.