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.     

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.     

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 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 .     

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 .     

EFFECTS OF RAW MATERIALS AND PROCESS VARIABLES ON THE HEAT PENETRATION TIMES, FIRMNESS, AND PECTIC ENZYME ACTIVITY OF DICED TOMATOES (HALLEY BOS 3155 CV)

The effects of raw materials and process variables on the heat penetration times into diced tomatoes (Halley Bos 3155 cv) were evaluated. Variables included dice size (1.27 and 2.54 cm), maturity at harvest (red and red+2 weeks), and processing temperature (88 and 92C). Heat penetration times between dice sizes were significantly different, but not between maturities or processing temperatures. Tomatoes were also evaluated for firmness, pectin-methylesterase (PME) and polygalacturonase (PG) activities. Half-inch size diced tomatoes were processed at 88 and 92C, and evaluated for firmness using the shear-compression method. Firmness decreased to 60% of the initial raw firmness from 8.8 × 10⁵ to 5.3 × 10⁵ g-mm after 15 s at 88C, and to 50% from 8.8 × 10⁵ to 4.4 × 10⁵ g-mm after 15 s at 92C. Diced tomato firmness showed a slight firming trend after 150 s at both temperatures. PME was inactivated after 45 s, while 5% residual PG activity remained after 3 min.     

MOLD GROWTH AND PRESENCE OF AFLATOXIN IN SOME TURKISH CHEESES

Twenty-five random fresh market samples of Van herby cheese and pickled white cheese were examined for molds and aflatoxins. The mean total mold count in Van herby cheese was 2.50 × 10⁵/g; in pickled white cheese it was 4.95 × 10⁴/g. The mycoflora on the cheeses were determined. In all cheeses, over 65% of molds were Penicillium species . Aspergillus made up 0 to 1.6 % and 2.6 % to 4.0 % of the mold on pickled white cheese and Van herby cheese, respectively. Other isolated molds belonged to Mucor, Geotrichum and Trichoderma genera. None of the samples contained aflatoxins and none of the 6 Aspergillus isolates was an aflatoxin producer .     

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.     

Effect of technological parameters on the characteristics of kasseri cheese made from raw or pasteurized ewes' milk

Two groups of kasseri cheese (pasta filata type) were manufactured from raw or pasteurized ewes' milk, without starter cultures. Cheeses of each group were divided into two subgroups: the first was ripened and stored at 4°C and packaged in plastic film; the second ripened and stored at 15°C and coated with paraffin wax. Milk pasteurization and technological parameters had a significant effect on the pH ( P  < 0.05), while only technological parameters had an effect on the total solids content. At day 120, the range of mean cfu/g counts for the mesophilic aerobic flora was 9.5 × 10⁷−1.4 × 10⁸; for the thermophilic streptococci, the range was 2.6 × 10⁷−7.6 × 10⁷; and for the thermophilic bacilli, 9.8 × 10⁶−1.7 × 10⁷. Changes in the N fractions became significant after 30 days of ripening. For mature 120-day-old cheeses, the percentage of total N soluble at pH 4.6 was 22.7%–22.9% in raw milk cheeses and 19.0%–21.7% in pasteurized milk cheeses. The percentage of total N soluble at 12% TCA was 10.1%–12.2% in raw milk cheeses and 7.3%–11.5% in pasteurized milk cheeses; the percentages of total N soluble at 5% PTA were 3.1%–4.0% and 2.6%–3.6%, respectively. The residual α_s-casein percentages at day 120 ranged between 63% and 78% of the respective area at day 1; the residual β-casein ranged between 67% and 75%. There were some characteristic differences in the reverse phase-HPLC peptide profiles of the four cheeses. In general, the effect of the different ripening conditions was more pronounced in cheeses made from pasteurized milk.     

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 .     

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     

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.     

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.     

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.     

Influence of immersion freezing in NaCl solutions and of frozen storage on the viscoelastic behavior of mozzarella cheese

ABSTRACT:  The freezing of Mozzarella cheese by immersion in NaCl solutions may be an innovative procedure for the dairy industry because it combines conveniently salting and freezing processes. In this work, the influence of this type of freezing method and of the frozen storage of samples on the viscoelastic behavior of Mozzarella cheese was studied. Slabs (2 × 10 × 10 cm³) were immersed in 23% w/w NaCl solutions (control samples: 4 °C, 90 min; frozen samples: −15 °C, 180 min). Half of the frozen samples were immediately thawed at 4 °C . The other half was stored at −20 °C for 2 mo and then was thawed at 4 °C (frozen-stored samples). Samples were stored at 4 °C and assayed at 1, 7, 14, 20, 27, 34, and 41 d. Rheological tests were carried out in oscillatory mode (parallel-plate geometry, diameter: 20 mm, gap: 1 mm, frequency: 1 Hz). Strain sweeps were run (0.001 ≤γ₀≤ 0.1) at 20, 40, and 60 °C, and temperature sweeps were run from 20 to 65 °C (1.33 °C/min, γ₀= 0.005). Similar crossover temperatures were observed after 20 d of ripening. The influence of temperature on complex viscosity was studied by an Arrhenius-type equation. Activation energy values of 15.9 ± 0.4, 14.1 ± 0.5, and 13.8 ± 0.6 kcal/mol were obtained at 41 d for control, frozen, and frozen-stored samples, respectively. Although the immersion freezing of Mozzarella cheese affects some of the studied parameters, the differences observed between frozen and frozen-stored samples with control samples were small. Therefore, it was considered that the immersion freezing might be useful for the manufacture and commercialization of Mozzarella cheese.     

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-STRUCTURE RELATIONSHIPS IN HEAT-INDUCED SOY PROTEIN GELS

Structural factors responsible for textural properties of heat-induced soy protein gels were investigated using microscopic and mechanical testing techniques. The gels were prepared by heating 20% soy protein pastes for 30 min at temperatures ranging from 25 to 130°C . Gel hardness increased linearly with the heating temperature up to 80°C , and decreased when the gels were heated at over 90°C , especially over 120°C . The equilibrium modulus estimated by tensile stress relaxation experiments was of the order of 10⁴ - 10⁵ dyne/cm², suggesting the presence of crosslinks, and there was a good correlation between the equilibrium modulus and the hardness of the gels. Solubility in phosphate buffer containing 2 -mercaptoethanol and/or urea suggested that the gel network was formed through crosslinking of the disulfide-, hydrogen- and hydrophobic-bonding types, and that the textural properties were governed by the degree of the network formation controlled by the heating temperature. SEM images of the 80°C -induced hard gels revealed a porous structure having membranous walls of thin compact film. With the 40°C -induced soft gels, the formation of the porous structure was not yet adequate, while a partial collapse of this structure was observed in the 120°C -induced fragile gels.     

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.