HEAT PASTEURIZATION OF CRAB AND SHRIMP FROM THE PACIFIC COAST OF THE UNITED STATES: PUBLIC HEALTH ASPECTS

ABSTRACT— A study of the potential public health hazard presented by coagulase-positive staphy-lococci, salmonellae and Clostridium botulinum in the meat of Dungeness crab and Pacific Coast shrimp pasteurized in flexible plastic containers revealed potentially toxinogenic staphylococci on the commercial nonpasteurized product, in 15% of the shrimp and 9% of the crab samples. No salmonellae or Cl. botulinum were isolated from, respectively. 26 and 54 samples of shrimp or 74 samples of crab. Pasteurization for 1 min at 180°F destroyed large inocula (10⁷ and 10⁸ cells) of staphylococci and salmonellae introduced into packages of the products, but processing for 5 min at 180°F allowed some members of an inoculum containing 10³ spores of Cl. botulinum type E to survive. While storage at 40°F prevented the growth on crab and shrimp meat of all staphylococci and salmonellae tested, it permitted growth and toxin formation by Cl. botulinum type E after 30–40 days. No toxin could be detected in packages inoculated with type A and proteolytic B spores and held at 50°F or lower. A 0.1% dip of sodium benzoate, with or without fumaric acid, did not prevent growth and toxinogenesis by Cl. botulinum types A, proteolytic B or E. It was concluded that for complete safety a holding temperature of 36°F or lower at all times would be required, but that it could not be expected to be maintained in commercial channels.     

Postharvest Microbiology of Farm-reared, Tropical Freshwater Prawn (Macrobrachium Rosenbergii)

ABSTRACT: The microbiological quality of farm-reared, tropical freshwater prawn ( Macrobrachium rosenbergii ) stored at 2 different temperatures was studied. The prawn muscle was found to have the initial bacterial load of 10⁴ cfu/g. The lactics and vibrios were in the range of 10² cfu/g, while the E. coli , aeromonads, staphylococci, anaerobes, and molds were in the level of 10¹ cfu/g. Salmonella and Vibrio cholerae were present in the prawn muscle. The prawn muscle held at room temperature (28 ± 2 °C) was organoleptically acceptable up to 8 h, when the bacterial load was more than 10⁶ cfu/g. However, the prawn muscle stored at freezer temperatures (−10 to −15 °C) was found to be in acceptable condition even after 30 d of storage and the bacterial load was fluctuating in the range of 10³ to 10⁴ cfu/g.     

GROWTH OF STAPHYLOCOCCUS AUREUS AND ENTEROTOXIN A PRODUCTION IN FOODS CONTAINING POLYPHOSPHATES

Effects of pyro-, tripoly- and hexametaphosphates (0.5 and 1%, w/w) on growth of Staphylococcus aureus strain 196E and enterotoxin A (SEA) production were studied in cooked custard and beef at 22 and 30°C. No effect was observed in custard, where cell numbers/g increased from 10³ to 10⁸ and SEA reached 4.2 ng/g after 48 h at 22°C, irrespective of treatment. Cell numbers in cooked beef were ca. 10⁹/g after 48 h, and reduced numbers (by 1.5–2 log cycles) were found in samples containing 0.5 and 1% pyrophosphate during incubation at 22%, but not at 30°C. SEA concentrations in beef were 28 ng/g after 48 h at 22°C, and 93 and 184 ng/g after 24 and 48 h, respectively, at 30°C. SEA concentration correlated with amount of growth, and was nondetectable when cell numbers were ± 10⁶/g. Reduction of the meat pH by sodium acid pyrophosphate contributed to the observed inhibitory effect.     

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.     

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.     

The yeast flora of stored ready-to-use carrots and their role in spoilage

Spoilage of ready-to-use grated raw carrots packaged in polymeric films and stored at 10°C was investigated for involvement of yeasts. Cryptococcus albidus was only isolated during the first 3 days of storage, increasing to levels of 10⁵–10⁶g^-1. Candida lambica was more commonly isolated after 3–7 days of storage, and reached 10⁷–10⁸g^-1 after 12 days. Candida sake was present throughout storage, increasing from 10⁵–10⁶ after 3 days to 10⁷–10⁸ after 12 days. In some samples, Candida parapsilosis and Candida tropicalis were also isolated at levels similars to C. sake . All the yeasts isolated at the end of storage were fermentative species and their metabolism was characterized with a Warburg apparatus. Neither the number of yeasts nor the composition of the yeast flora were related to the deterioration of the product. Although Candida lambica inoculated on grated carrots caused spoilage after 12 days at 10°C, the high O₂ permeable film was most effective in reducing exudate.     

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 .     

PROBABILITY OF GROWTH OF CLOSTRIDIUM BOTULINUM AS AFFECTED BY STRAIN, CELL AND SEROLOGIC TYPE, INOCULUM SIZE AND TEMPERATURE AND TIME OF INCUBATION IN A MODEL BROTH SYSTEM

Using factorial design experiments and MPN methodology, we evaluated the probability (P) of growth initiation of 17 individual proteolytic (A,B,F) and non-proteolytic (B,E,F) C. botulinum strains (spores and cells) in a model broth as it is affected by strain, cell and serologic type, inoculum size (10⁰–10⁶), temperature (4–47°C) and time of incubation (up to 28 days). Regression analyses of the P of growth of the most capable strains for all conditions tested, allowed the development of quantitative equations relating P of growth initiation by one spore or cell to the variables studied. The close agreement between observed and predicted Ps demonstrated the potential usefullness of the modeling approach in studying microbial interactions with food environments.     

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.     

Factors controlling the growth of Clostridium botulinum types A and B in pasteurized, cured meats III. The effect of potassium sorbate

The growth of Clostridium botulinum types A and B spores, at 10¹ or 10³ per container, was studied in a pork slurry system containing nitrite (40 μg/g), sodium chloride (2.5, 3.5, 4.5% w/v) sodium isoascorbate (550 μg/g) at varying pH levels, with or without potassium sorbate (0.26% w/v), without heating and after two heat treatments (80°C for 7 min, and 80°C for 7 min + 70°C for 1 hr) followed by storage at 15, 17.5, 20 or 35°C for up to 6 months. At a given spore inoculum, potassium sorbate significantly decreased toxin production, as did increasing NaCl, decreasing pH or decreasing storage temperature. Heat treatment did not significantly affect spoilage or toxin production overall, but interacted significantly with some factors. The effect of sorbate was greater at 3.5% NaCl than at 2.5%, at pH values below 6.0, and at low storage temperature.     

Direct gas-fired heating of a pilot-scale spray drier using natural gas for milk powder manufacture

A pilot-scale spray drier was converted to direct gas-fired heating using natural gas to determine its effects on the chemical and physical properties of the skimmed milk powders dried by this technique. There were no significant differences in acidity, bulk density, solubility index and moisture content compared with the control samples ('indirect' heating of drier) during the course of 11 trials. The heated drying air (200°C) was analysed for the presence of NO + NO₂, CO and CO₂, and some increases at ppm levels in the nitrate (NO₃-) and nitrite (NO₂-) content of the milk powders were observed. Colour and flavour characteristics of the samples were not affected.     

Low Temperature Growth of Type E Clostridium botulinum Spores. 1. Effects of Sodium Chloride, Sodium Nitrite and pH

SUMMARY— The effects of pH and the addition of sodium chloride (NaCl) or sodium nitrite (NaNO₂) to trypticase-peptone-glucose (TPGI and trypticase-peptone-sucrose-yeast extract (TPSY) media upon spore outgrowth were investigated using seven Type E Clostridium botulinum strains. An inoculum of 1.0 × 10⁵ spores/ml was used and the pH was adjusted with hydrochloric acid to cover the range of 5.2 to 6.6. To define salt tolerance, NaCl was added to the media at intervals of 0.5% in the range of 2.0 to 5.0% and at 0.1% intervals in the 4.5 to 5.0% range. The effect of NaNO₂ was investigated with the addition of 100 and 200 ppm NaNO₂ to the media. Samples were incubated at 30, 15.6, 10, 7.2, 5.0 and 3.4°C. Outgrowth of all strains tested was inhibited at pH 5.2 at 15.6°C. Inhibition occurred at higher pH at lower temperatures. None of the strains showed outgrowth with 4.87% NaCl in the media at any of the incubation temperatures used. Addition of 100 and 200 ppm NaNO₂ to the media inhibited the outgrowth of the Minneapolis, 517, 26080 and A6247 strains but not the Kalamazoo and Seattle Forks strains.     

Osmotic Dehydration of Tomato in Sucrose Solutions: Fick's Law Classical Modeling

ABSTRACT:  Osmotic dehydration of tomato was modeled by the classical Fick's law including shrinkage, convective resistance at the interface and the presence of water bulk flow. Tomato slices having 8 mm thickness were osmotically dehydrated in sucrose solutions at 50, 60, and 70 °Brix and at 35, 45, and 55 °C. Other experiments were done in a 70 °Brix sucrose solution at 35 °C with tomato slices of 4, 6, and 8 mm thickness and at different motion levels (velocities 0, 0.053, and 0.107 m/s). Tomato weight, water content, and °Brix of the products were measured as a function of processing time (20, 40, 80, 160, and 320 min). Results showed that temperature, concentration, thickness, and solution movement significantly influenced water loss and sucrose gain during the osmotic dehydration of tomato. The model predicted the modifications of soluble solid content and water content as a function of time in close agreement with the experimental data. Experimental Sherwood number correlations for sucrose and water were determined as Sh _s = 1.3 Re ^0.5 Sc _s^0.15 and Sh _w = 0.11 Re ^0.5 Sc _w^0.5, respectively. The effective diffusion coefficients of water (4.97 10⁻¹¹– 2.10 10⁻¹⁰ m²/s) and sucrose (3.18 10⁻¹¹– 1.69 10⁻¹⁰ m²/s) depended only on temperature through an Arrhenius-type relationship.     

Mechanism of inactivation of heat-tolerant spores of Bacillus stearothermophilus IFO 12550 by Rapid Decompression

The effect of three rapid decompression methods to clear the mechanism of inactivation of heat tolerant spores of Bacillus stearothermophilus IFO 12550 was investigated. Pressurization of the spores at 200 MPa and 75°C for 60 min caused a kill of 10⁴ CFU/mL by the link-motion system but the nonrotational rod valve and E.G. seal methods gave a kill of about 10³ CFU/mL. Sterilization was due to the physical breakdown of spore coat, and was induced by its physical permeability of water at high pressure and temperature. Rapid decompression by the link-motion system at 200 MPa decreased the D-value of the spores from 3000 min (100°C, one atmosphere) to 6 min, 11 min, and 17 min at 95, 85, and 75°C, respectively.     

APPLICATION OF CHLORINE TO REDUCE POPULATIONS OF ESCHERICHIA COLI ON BEEF

The effects of chlorine against 2 strains of E. coli attached to the surface of beef carcass tissue (BCT) were examined using a model carcass washer. Lean and adipose BCT with approximately 5 log ₁₀ CFU/cm ² E. coli bacteria were spray-treated with water and sodium hypochlorite (NaOCl) to give chlorine concentrations of 50, 100, 250, 500, or 800ppm, incubated for 24 h, 4C, and E. coli populations enumerated. Spray treatments with water did significantly (P < 0.05) reduce the bacterial populations of either organism attached to lean or adipose BCT, as compared to populations of controls; however, reductions were less than 0.60 log ₁₀ CFU/cm ². Treatments with 500 and 800 ppm chlorine against E. coli ATCC 25922 attached to BCT resulted in the greatest reductions of 1.22 and 1.28 log ₁₀ CFU/cm ², respectively. At 800 ppm chlorine , E. coli O157:H7 ATCC 43895 attached to BCT was reduced by 1.04 log ₁₀ CFU/cm ², whereas spray treatments with 50, 100, 250, and 500 ppm chlorine resulted in reductions of < 1 log ₁₀ CFU/cm ². Spray treatments with chlorine from sodium hypochlorite solutions reduced populations of E. coli, however, these reductions were not sufficient to completely inactivate the bacteria attached to red meat .     

EARLY DETECTION OF CLOSTRIDIUM PERFRINGENS ENTEROTOXIN AND ITS RELATIONSHIP TO SENSORY QUALITY OF COOKED CHICKEN

Growth, sporulation, and enterotoxin formation by Clostridium perfringens NCTC 8239 were determined in chicken thigh meat incubated at 45°C for 1.5 h and 37°C for up to 12.5 h. With an inoculum of 10⁶ vegetative cells per g, the cell counts reached mean log ₁₀ 7.32/g after 6 h of incubation and remained in that range through 14 h. Heat-resistant spores (log₁₀ 2.48/g) were first detected at 4 h, and the number increased to log₁₀ 5.19/g at 14 h. Enterotoxin (0.19 μg/g) was first detected after 2 h of incubation (1.5 h at 45°C and 0.5 h at 37°C) in the absence of detectable sporulation, and the enterotoxin concentration increased to 0.76 μg/g after 14 h. Significant differences (p < 0.01) in the odor, color, and texture scores for inoculated versus uninoculated cooked chicken following 2 h incubation correlated with the production of enterotoxin and suggested that these parameters could be used as indices of chicken spoilage by C. perfringens.     

THE SURVIVAL OF MARINE VIBRIOS IN MERCENARIA MERCENARIA, THE HARDSHELL CLAM

A depuration chamber was used to study the persistence of marine vibrios in the hardshell clam, Mercenaria mercenaria. Specimens of M. mercenaria were incubated for two h in artificial seawater containing 10³ cells/ml each of the following bacterial species; Vibrio parahaemolyticus, Vibrio harveyi and Escherichia coli, and then transferred to the depuration chamber (a tank through which U. V.-sterilized artificial seawater was continually flowing). Numbers of the three bacterial species in tissues of M. mercenaira removed from the chamber at various times were determined by differential plating techniques. The number of each species ranged from 10² to 10³ colony-forming units/gram tissues immediately after transfer to the depuration chamber. After 24 h at 25°C the number of E coli cells detected had decreased over 100-fold. Generally, V. parahaemolyticus and V. harveyi were found in increased abundance after 24 h. The abundance of V. parahaemolyticus and V. harveyi in clams that had been incubated in the depuration chamber for 72 h at 25°C was approximately 10% of the abundance of these species immediately after transfer to the chamber. Similar results were obtained when the incubation temperature was 8 or 15°C and when initial cell concentrations were altered. Thus, V. harveyi and the potential human pathogen, V. parahaemolyticus which are both of marine origin were not removed from M. mercenaria at a rate comparable to the rate at which M. mercenaria depurated cells of E. coli.     

Inactivation of Escherichia coli O157:H7 and Salmonella enteritidis in Liquid Egg White Using Pulsed Electric Field

ABSTRACT: The effects of temperature and pulsed electric field (PEF) intensity on inactivation of pathogens such as Escherichia coli O157:H7 and Salmonella enteritidis in egg white was investigated. Liquid egg white inoculated with 10⁸ colony-forming units (CFU)/mL of each pathogen was treated with up to 60 pulses (each of 2 JAS width) at electric field intensities of 20 and 30 kV/cm. The processing temperatures were 10°C, 20°C, and 30°C. After treatment, uninjured and total viable cells were enumerated in selective and nonselective agars, respectively. Maximum inactivations of 3.7 and 2.9 log units were obtained for S. enteritidis and E. coli O157:H7, respectively, while injured cells accounted for 0.5 and 0.9 logs for E. coli O157:H7 and S. enteritidis , respectively. For both bacteria, increasing treatment temperature tended to increase the inactivation rate. There was synergy between electric field intensity and processing temperature. The inactivation rate constant k _T values for E. coli O157:H7 on both selective and nonselective agars were 8.2 × 10^-3 and 6.6 × 10^-3/μS, whereas the values for S. enteritidis were 16.2 × 10^-3 and 12.6 × 10^-3/μS, respectively. The results suggest that E. coli O157:H7 was more resistant to heat-PEF treatment compared with S. enteritidis.     

The air drying behaviour of fresh and osmotically dehydrated banana slices

Ripe banana, cut to 10mm thick slabs were osmotically treated in sugar solutions of 35, 50 and 65° Brix for 36h. The initial moisture content fell from a value of 3.13kg H₂O DM to 2.19, 1.63 and 1.16kg H₂O kg⁻¹ for treatment in the three solutions, respectively. These slabs, with Total Soluble Solids (TSS) contents of 26, 34 and 39° Brix, respectively, as well as freshly cut but untreated slabs (15° Brix) were air dried in a cabinet type tray drier to near equilibrium conditions at fixed temperatures from 40 to 80°C and at a constant air speed of 0.62m s⁻¹. Drying was found to occur in the falling rate period only for both banana types and two drying constants K₁ and K₂ were established for a first and second falling rate period of drying. Increasing the drying air temperature significantly enhanced the drying rate and the K-values, except at 80°C when the rates fell, possibly because of case hardening of the slabs. Reducing the slab thickness also improved the drying rate, but increasing the air speed to 1.03m s⁻¹ did not have any profound effect. As the sugar content of the banana slabs increased through the osmotic treatment, drying rates fell. Calculated apparent moisture diffusivities at 60°C ranged from 34.8× 10⁻¹⁰ m² s⁻¹ (fresh slab) to 8.8×10⁻¹⁰ m² s⁻¹ for dried (39° Brix) slabs. The moisture diffusivity was significantly lowered as the moisture content dropped in drying and with increased levels of sugar. Previously osmosed and then air dried banana slabs showed appealing colour and texture compared to the fresh banana.     

Decontamination Efficacy of Combined Chlorine Dioxide with Ultrasonication on Apples and Lettuce

ABSTRACT:  The bacterial reduction of Salmonella and Escherichia coli O157:H7-inoculated apples and lettuce by ClO₂ at 0, 5, 10, 20, and 40 ppm with and without 170-kHz ultrasonic treatments for 3, 6, and 10 min, respectively, have been studied. The treatments of ClO₂ at 20 and 40 ppm for 3, 6, and 10 min or at 5 and 10 ppm for 6 and 10 min with 170-kHz ultrasonication caused 3.115 to 4.253 log reductions in Salmonella and 2.235 to 3.865 log reduction in E. coli O157:H7 on inoculated apples. Using combined ClO₂ and ultrasonication to treat 4.48 × 10⁴ CFU/g Salmonella and 1.07 × 10⁵ CFU/g E. coli O157:H7-inoculated lettuce, the bacterial reductions were 2.257 to 2.972 and 1.357 to 2.264 log, respectively. The residual ClO₂ decreased with increasing treatment times, over 80% of ClO₂ was detected after the 3-min treatment, and more than 70% remained after the 10-min treatment time. No bacteria were recovered from the posttreatment solutions of ClO₂ or ClO₂ combined with ultrasonication. The temperature of the ClO₂ treatment was 20.1 °C, and it increased to 40.1, 44.9, and 50.3 °C, with 170-kHz ultrasonic treatments for 3, 6, and 10 min, respectively, on apples.