Effect of the Parabens With and Without Nitrite on Clostridium botulinum and Toxin Production in Canned Pork Slurry

Antimiciobials were evaluated in thioglycollate broth at pH 6.5 for the ability to inhibit growth and toxin production by C. botulinum 12885A and ATCC 7949 (Type B). Methyl, ethyl, propyl, and butyl parabens (0.1%) and sorbic acid (0.2%) were effective in inhibiting growth of C. botulinum 12885A and ATCC 7949 in broth. Ethyl, propyl, and butyl parabens (0.1%) and sorbic acid (0.2%) inhibited toxin production by both strains in culture medium. Ethyl, propyl, butyl parabens (0.1%) and sorbic acid (0.2%) were individually added to a comminuted pork slurry having salt and sugar, with or without 40 ppm sodium nitrite. Cans were inoculated with a mixture of C. botulinum 12885A and ATCC 7949 spores. The canned product was abused by holding at 27°C and was observed over a 3-month period for swollen cans. Swollen cans were examined for botulinal toxin by the mouse bioassay. Propyl and butyl paraben did not inhibit or delay toxin production. Ethyl paraben with or without nitrite delayed toxin production for 4 wk. Sorbic acid inhibited toxin for 3 wk; when 40 ppm nitrite was added to the sorbic acid treatment, toxin production was delayed for 4 wk.     

Effect of Sodium Acid Pyrophosphate in Combination with Sodium Nitrite or Sodium Nitrite/Potassium Sorbate on Clostridium botulinum Growth and Toxin Production in Beef/Pork Frankfurter Emulsions

Combinations of sodium acid pyrophosphate (SAPP) with added sodium nitrite and/or potassium sorbate were tested at various pH levels to determine effectiveness in delaying Clostridium botulinum growth and toxin production in frankfurter emulsions. Formulations containing sodium nitrite (40 ppm), potassium sorbate (0.26%) and SAPP (0.4%) resulted in a greater delay of toxin production (12–18 days) than other combinations (6–12 days) having similar pH values. Treatments containing 0.4% SAPP appeared to be more inhibitory than their counterparts without SAPP, displaying less numbers of toxic samples during the 53-day storage period at 27°C. Aerobic mesophilic colony counts and residual nitrite data showed little difference among treatments.     

EFFECT OF NITRITE AND STORAGE TEMPERATURE ON THE ORGANOLEPTIC QUALITY AND TOXINOGENESIS BY Clostridium botulinum IN VACUUM-PACKAGED SIDE BACON

The effect of nitrite and storage temperature and toxinogenesis by Clostridium botulinum in vacuum-packed side bacon was investigated. In two series of experiments (A & B) bacon packs were prepared with levels of 0, 50, 100, 150 and 200 ppm nitrite and inoculated with C botulinum at 10² spores/g and 10⁴ spores/g. Packs A were incubated at 20 and 30° C and packs B at 30°C only. Both were held for a maximum of 32 days and analyzed for toxin at intervals of 2, 4, 8, 16 and 32 days. At 20°C none of the controls without nitrite was found to be toxic after 32 days. At 30°C inhibition of toxin formation at the higher nitrite levels was observed at 32 days. Organoleptic evaluation of the bacon packs stored at 30° C showed about one-third of the toxic samples examined were acceptable to the panel.     

Antibotulinal Activity of Methyl and Ethyl Fumarates in Comminuted Nitrite-Free Bacon

Mono- and dimethyl and ethyl esters of fumaric acid were evaluated for their antibotulinal efficacy in cans of comminuted nitrite-free bacon. At 0.125%, all were equal or superior to 120 ppm nitrite in preventing toxin formation in cans incubated at 30°C for 8 wk. No cans swelled or became toxic when mono- or dimethyl fumarate was added. With monoethyl fumarate, two cans out of twenty swelled but were nontoxic. The least effective ester was diethyl fumarate but its activity was equal to that of 120 ppm nitrite.     

Mathematical modeling of growth of non-O157 Shiga toxin-producing Escherichia coli in raw ground beef

Abstract: The objective of this study was to investigate the growth of Shiga toxin‐producing Escherichia coli (STEC, including serogroups O45, O103, O111, O121, and O145) in raw ground beef and to develop mathematical models to describe the bacterial growth under different temperature conditions. Three primary growth models were evaluated, including the Baranyi model, the Huang 2008 model, and a new growth model that is based on the communication of messenger signals during bacterial growth. A 5 strain cocktail of freshly prepared STEC was inoculated to raw ground beef samples and incubated at temperatures ranging from 10 to 35 °C at 5 °C increments. Minimum relative growth (<1 log₁₀ cfu/g) was observed at 10 °C, whereas at other temperatures, all 3 phases of growth were observed. Analytical results showed that all 3 models were equally suitable for describing the bacterial growth under constant temperatures. The maximum cell density of STEC in raw ground beef increased exponentially with temperature, but reached a maximum of 8.53 log₁₀ cfu/g of ground beef. The specific growth rates estimated by the 3 primary models were practically identical and can be evaluated by either the Ratkowsky square‐root model or a Bělehrádek‐type model. The temperature dependence of lag phase development for all 3 primary models was also developed. The results of this study can be used to estimate the growth of STEC in raw ground beef at temperatures between 10 and 35 °C. Practical Application: Incidents of foodborne infections caused by non‐O157 Shiga toxin‐producing Escherichia coli (STEC) have increased in recent years. This study reports the growth kinetics and mathematical modeling of STEC in ground beef. The mathematical models can be used in risk assessment of STEC in ground beef.     

Compositional Factors that Influence Lipid Peroxidation in Beef Juice and Standard Sausages

In order to identify how different additives influenced lipid peroxidation formation, a sausage only using beef juice as pigment source and a standard beef–pork meat sausage were studied. The effects of different additives, including fish oil, myoglobin, nitrite, clove extract, and calcium sources on oxidation and sensory properties were examined. Both sausage systems were stored in 3 different manners prior to testing: (1) frozen immediately at ?80 °C; (2) chilled stored for 2.5 weeks followed by fluorescent light illumination at 4 °C for another 2 wk; (3) frozen at ?20 °C for 5 mo. The frozen group 3 showed the highest peroxide formation and thiobarbituric acid reactive substances (TBARS) for both sausage systems. Unpolar peroxides dominated in both systems. The clove extract could offset the peroxide formation from myoglobin/beef juice and/or fish oil, but the addition of clove flavor was recognized by the sensory panelists. Calcium addition reduced lipid peroxide formation. Added nitrite and fish oil seemed to interact to stimulate nitroso‐myoglobin formation. Nitrite was identified to interact with clove addition and thereby, relatively speaking, increased TBARS. The 2 sausage systems generally ranked the additives similarly as pro– and antioxidants.     

Listeria monocytogenes Survival Model Validated in Simulated Uncooked-Fermented Meat Products for Effects of Nitrite and pH

Previous modeling studies in broth cultures demonstrated that acidity and nitrite increased the inactivation rate of Listeria monocytogenes. To validate this effect during storage of simulated uncooked-fermented meat products, lean beef was ground with salt, adjusted to pH 4.0–5.1, and treated with nitrite at 0–300 μg/mL. Samples were immediately inoculated with L. monocytogenes (10⁷ CFU/g) and survivors were enumerated over 21 days storage at 37°C. The time to achieve a four log decline as greatly affected by pH, ranging from 21 days at pH 5.0 to < 1.0 day at pH 4.0. Growth occurred at pH 5.1 after a long lag period. Nitrite additions did not affect survival, suggesting that the effective concentration was the rapidly decreasing residual nitrite level.     

Factors controlling the growth of Clostridium botulinum types A and B in pasteurized, cured meats

The effect of combinations of sodium chloride (2.5, 3.5, 4.5% w/v on water), sodium nitrite (100, 200, 300 μg/g), sodium nitrate (0, 500 μg/g), sodium isoascorbate (0,1000 μg/g, or equimolar with nitrite level) and polyphosphate (Curaphos 700; 0, 0.3% w/v), on the growth of Clostridium botulinum types A and B was studied in an experimental pork slurry system, without heating and after two heat treatments (80°C for 7 min and 80°C for 7 min plus 70°C for 1 hr) followed by storage at: 15, 17.5, 20 or 35°C for up to 6 months.
Statistical analyses showed that increasing salt or nitrite levels, adding isoascorbate or nitrate, using the highest heat treatment or decreasing the storage temperature all significantly reduced toxin production by Cl. botulinum . The addition of 0.3% polyphosphate (Curaphos 700) significantly increased toxin production. There were many significant two-factor interactions; the effect of increasing nitrite was relatively less if isoascorbate was present, at 4.5% salt, or at low storage temperature. The presence of isoascorbate also counteracted the increase in toxin production attributed to the presence of polyphosphate.     

Dietary Plant Sterols Supplementation Increases In Vivo Nitrite and Nitrate Production in Healthy Adults: A Randomized,Controlled Study

A randomized, double‐blinded, placebo‐controlled and crossover study was conducted to simultaneously measure the effects, 3 h after consumption and after 4‐wk daily exposure to plant sterols‐enriched food product, on in vivo nitrite and nitrate production in healthy adults. Eighteen healthy participants (67% female, 35.3 [mean] ± 9.5 [SD] years, mean body mass index 22.8 kg/m²) received 2 soy milk (20 g) treatments daily: placebo and one containing 2.0 g free plant sterols equivalent of their palmityl esters (β‐sitosterol, 55%; campesterol, 29%; and stigmasterol, 23%). Nitrite and nitrate concentrations were measured in the blood plasma and urine, using stable isotope‐labeled gas chromatography‐mass spectrometry. L‐arginine and asymmetric dimethylarginine concentrations in blood serum were measured using commercially available enzyme immunoassays. Nitrite and nitrate concentrations in blood plasma (nitrite 5.83 ± 0.50 vs. 4.52 ± 0.27; nitrate 15.78 ± 0.96 vs. 13.43 ± 0.81 μmol/L) and urine (nitrite 1.12 ± 0.22 vs. 0.92 ± 0.36, nitrate 12.23 ± 1.15 vs. 9.71 ± 2.04 μmol/L) were significantly elevated after 4‐wk plant sterols supplementation Placebo and 3‐h treatments did not affect the blood plasma and urinary concentrations of nitrite and nitrate. Circulating levels of L‐arginine and asymmetric dimethylarginine were unchanged in the placebo and treatment arms. Total plant sterols, β‐Sitosterol, campesterol, and stigmasterol concentrations were significantly elevated after 4‐wk treatments compared to the placebo and 3‐h treatments. Blood plasma nitrite and nitrate concentrations correlated significantly with the plasma total and specific plant sterol concentrations. Our results suggest that dietary plant sterols, in the combination used, can upregulate nitrite, and nitrate production in vivo.     

Impact of nitrite on detection of Listeria monocytogenes in selected ready-to-eat (RTE) meat and seafood products

ABSTRACT:  The impact of sodium nitrite (NaNO₂) on detection and recovery of Listeria monocytogenes from select ready-to-eat (RTE) foods including smoked salmon, smoked ham, beef frankfurters, and beef bologna was assessed. Nitrite-containing (NC; 100 to 200 ppm NaNO₂) or nitrite-free (NF) foods were inoculated with a 5-strain cocktail of L. monocytogenes by immersion into Butterfield's buffer solution containing 5.4 to 7.4 × 10³ L. monocytogenes per milliliter. Inoculated products were vacuum-packaged and stored at 5 °C. A weekly comparative analysis was performed for presence of L. monocytogenes using 5 detection methods on products held at 5 °C for up to 8 wk. L. monocytogenes initially present at <100 CFU/g during the first 2 wk of storage increased throughout the study, attaining final populations of approximately 1 × 10⁴ to 1 × 10⁵ CFU/g. Lactic acid bacteria predominated throughout the study in all products. Exposure to NaNO₂ (100 to 200 ppm) resulted in 83% to 99% injury to the L. monocytogenes strains tested. The genetic-based BAX^® System (DuPont™ Qualicon, Wilmington, Del., U.S.A.) and modified USDA/FSIS methods detected 98% to 100% of Listeria -positive food samples and were consistently superior to and significantly different ( P < 0.05) from conventional cultural methods in recovering Listeria from NC samples. Data show that nitrite-induced injury adversely affects detection and recovery of L. monocytogenes from NC food, confirming earlier findings that nitrite-induced injury masks L. monocytogenes detection in NC RTE food products. Nitrite-injured Listeria can subsequently repair upon nitrite depletion and grow to high levels over extended refrigerated storage.     

Microbiology of a Ham-Type Product Made from Soy-Extended Cured Beef and Inoculated with Clostridium sporogenes PA3679

The microbiological stability of a new ham-type product made from soy-extended cured beef was examined under wholesale (2–4°C) and retail (5°C) refrigerated storage conditions, and during abuse-temperature holding for 24 and 48 hr at 24-25°C after inoculation with Closrridium sporogenes PA3679. No microbiological effects (P >0.05) could be attributed to the level of soy protein isolate in the injection brine (0, 5, 7.5 or 9%) on the basis of mesophilic, psychrotrophic, anaerobic and lactic bacterial counts. Samples held at 2-4°C did not exceed 10⁴ CFU/g after 4 wk for any bacterial type examined. Product shelf-life was > 3 wk at 5°C and 2 wk at 8–10°C. Inoculated PA3679 did not grow in the product during 24 or 48 hr simulated mishandling (24-2°C).     

CONTROL OF CLOSTRIDIUM PERFRINGENS IN A MODEL ROAST BEEF BY SALTS OF ORGANIC ACIDS DURING CHILLING1

Control of Clostridium perfringens germination and outgrowth by the following salts of organic acids, sodium lactate [Purasal?S/SP (Purasal); 1.50, 3.00 and 4.80%], sodium lactate supplemented with sodium diacetate [Purasal? Opti.form? (Optiform), 1.50, 3.00 and 4.80%], buffered sodium citrate [Ional? (Ional), 0.75, 1.00 and 1.30]) and buffered sodium citrate supplemented with sodium diacetate [Ional Plus? (Ional Plus), 0.75, 1.00 and 1.30%] was evaluated during continuous chilling of a model roast beef product. Beef rounds were ground through an 1/8′’ plate and NaCl, potato starch and potassium tetra pyrophosphate were added to final concentrations of 0.85, 0.25 and 0.20%, respectively, and mixed. Portions (250 g) of the meat were mixed with either Purasal (1.5, 3.0 or 4.8%), Optiform (1.5, 3.0 or 4.8%), Ional (0.75, 1.0 or 1.3%) or Ional Plus (0.75, 1.0 or 1.3%) along with a control that did not have any added antimicrobials. Each product (10 g) inoculated with C. perfringens spores (ca. 2.2 log₁₀ spores/g) was packaged into vacuum bags (2 in. × 3 in.), vacuum sealed, heated to 60C within 1 h, and subsequently chilled from 54.4C to 7.2C in 18 or 21 h following exponential chilling rates. Products were sampled immediately after cooking to enumerate the C. perfringens populations (spores surviving heat treatment) and subsequent to chilling (total C. perfringens populations, including spores and vegetative cells resulting from germination and outgrowth of the spores). Chilling of cooked, model ground roast beef resulted in germination and outgrowth of C. perfringens spores; the population densities increased by 4.13 and 4.40 log₁₀ CFU/g, following 18 and 21 h chill rates, respectively. Incorporation of Purasal (1.5–4.8%), Optiform (1.5–4.8%), Ional and Ional Plus (0.75–1.3%) substantially (P ± 0.05) inhibited germination and outgrowth of C. perfringens spores. Incorporation of antimicrobial ingredients resulted in ± 1.0 log₁₀ CFU/g increase of the pathogen, except for model roast beef with Ional Plus at 0.75% concentration, following 18 h chilling rate. Similar results were obtained when 21 h chilling rate was followed, with roast beef containing ingredients (at all the concentrations) resulting in either reductions or ± 1.0 log₁₀ CFU/g growth in total C. perfringens populations, except for Purasal and Ional Plus at 1.5 and 0.75% concentrations, respectively. Use of sodium salts of organic acids in formulation of model roast beef can reduce the risk of C. perfringens spore germination and outgrowth during extended chilling rates.     

Injury,Inhibition and Inactivation ofEscherichia coli O157:H7 by Potassium Sorbate and Sodium Nitrite as Affected by pH and Temperature

The effect of potassium sorbate (0–2 g litre⁻¹) and sodium nitrite (0–1 g litre⁻¹) on the growth of four strains of Escherichia coli O157: H7 in tryptic soya broth at various pH levels (pH 4·0–7·0 for sorbate, pH 5·0–8·0 for nitrite) were determined at 37°C and 4°C. Among the pH levels tested, sorbate and nitrite exhibited the highest antimicrobial activity at pH 4·0 and 5·0, respectively. At pH 5·0 and 37°C, the presence of 500 mg litre⁻¹ sorbate or 200 mg litre⁻¹ nitrite completely inhibited the growth of E coli O157: H7. While at higher pH levels, 2 g litre⁻¹ sorbate or 1 g litre⁻¹, nitrite, the highest concentration tested, did not show significant antimicrobial action against the test organisms. At 4°C and pH 5·0, the inoculated test organisms did not showed any significant growth in preservative-free control media. Different degree of inactivation and injury was observed when E coli O157: H7 strain 933 was stored in TSB (pH 5·0) containing 1 g litre⁻¹ sorbate or nitrite at 37°C. At 4°C, inactivation and injury of E coli O157: H7 cells was not observed in the medium containing sorbate or nitrite throughout the 24 h experimental period.     

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.     

The effects of heating and chemical treatment on the haem and non-haem iron content of heat-induced porcine blood curd

The effect of heating temperature (40–80°C), time (0–50 min), additions of ascorbic acid (0–5 g litre^?1) and sodium nitrite (0–200 mg litre^?1) on total, non-haem and haem iron of heat-induced porcine blood curd (a famous edible blood food in Taiwan) was investigated. The results show that non-haem iron content increased significantly (P < 0.05) when heated at above 55°C and was enhanced linearly (r = 0.96) with heating time at 80°C, while haem iron decreased relatively. In addition, a non-haem iron increase was observed in ascorbic acid, the maximum found with treatment at 4 g litre^?1. On the contrary, haem iron content increased with the presence of sodium nitrite and there was a significant change with addition above 50 mg litre^?1.     

Effects of Sodium Chloride Reduction and Polyphosphate Addition on Clostridium Botulinum Toxin Production in Turkey Frankfurters

Mechanically deboned turkey meat emulsions were made with 1.0, 1.5, 2.0, 2.5, or 3.0% salt (NaCl), or with combinations of 1.5% or 2.0% salt with 0.4% sodium tripolyphosphate (TPP), sodium hexametaphosphate (HMP), or sodium acid pyrophosphate (SAPP). Sodium nitrite levels were constant at 150 ppm. Emulsions were inoculated with a mixture of 10 strains of C. botulinum (10³/g) and incubated at 27°C. Increasing NaCl content from 1.0% to 3.0% delayed toxin production by 3 days on the average. Toxin production was detected earlier when TPP was added, HMP had no effect, and SAPP delayed toxin production.     

Inhibition of Clostridium botulinum by Antioxidants and Related Phenolic Compounds in Comminuted Pork

Antioxidants, esters of p-hydroxybenzoic acid and gallic acid, and related phenolic compounds were evaluated for their activity against growth and toxin production of C. botulinum types A and B in comminuted pork. With an inoculum level of 8,000 spores/g of meat most of the chemicals at a concentration of 1,000 ppm delayed first swell formation for less than 3 days beyond the control. For most compounds, the time to first swell formation beyond that of the control increased as the spore level per g of meat decreased. 8-Hydroxyquinoline at a concentration of 200 ppm or in combination with sodium nitrite (40 ppm) inhibited the growth and toxin production of C. botulinum in comminuted pork for 60 days at 27°C. 8-Hydroxyquinoline was more active in inhibition of growth and toxin production of C. botulinum in comminuted pork than in prereduced Thiotone-yeast extract-glucose broth.     

Evaluation of liquid smoke treated ready-to-eat (RTE) meat products for control of Listeria innocua M1

ABSTRACT:  Liquid smoke fractions (S1, S2, S3, and S4) were applied on ready-to-eat (RTE) meat products to control the growth of inoculated Listeria innocua M1. Turkey rolls and roast beef products were dipped in liquid smoke, surface inoculated with L. innocua M1 (10² CFU/25 cm² RTE meat surface), vacuum packaged, and stored at 4 °C. Section 8.5 of USDA's detection and isolation procedure for L. monocytogenes was employed in conjunction with a Micro-ID™ system for L. innocua M1 identification (ID). Products treated with smoke fractions S1, S2, and S3 were negative for L. innocua M1 at 2 and 4 wk during incubation at 4 °C. Products treated with S4 were positive for L. innocua M1 immediately following inoculation and after storage for 2 and 4 wk. Smoke fractions S1, S2, and S3 exhibited pH values lower than 4.6, acidity values higher than 1.5%, and carbonyl concentrations higher than 110 mg/mL. All liquid smoke fractions contained similar phenol concentrations (0.3 to 0.6 mg/mL), suggesting that phenols may have a limited role in the bactericidal effects of liquid smoke fractions against specific microorganisms.     

Prevention of Clostridium botulinum Type A, Proteolytic B and E Toxin Formation in Refrigerated Pea Soup by Lactobacillus plantarum ATCC 8014

The ability of Lactobacillus plantarum ATCC 8014 to inhibit Clostridium botulinum toxin production in pea soup was investigated. Soup containing C. botulinum spores (10³/g) with and without L. plantarum (10⁶/g) were evaluated. Soup containing only type A spores was toxic on days 1 and 2 when incubated at 35°C and 25°C, respectively. Soup containing only proteolytic type B spores was toxic on days 2 and 5 at 35°C and 25°C, respectively. Soup containing only type E spores was toxic at 25°C, 15°C, and 5°C in 7, 7, and 63 days respectively. No toxin was found in soup containing C. botulinum spores plus L. plantarum at any temperature studied.     

Thermal Inactivation of Clostridium botulinum Toxin Types F and G in Buffer and in Beef and Mushroom Patties

The time-temperature relationships for the inactivation of crude toxins prepared from Clostridium botulinum type F strain Wall 8 and type G strain G89 were determined in three different heating menstrua. Toxins dilated to 7,700–32,000 mouse LD₅₀ units/0.5 ml in beef and mushroom patties (pH 6.05), in 0.1M phosphate buffer (pH 6.05), and in 00.1M acetate buffer (pH 5.0) were heated at temperatures from 65.6–76.7°C. In the beef patties, inactivation times ranged from an average of 108.1–1.6 min for type F toxin, and from an average of 158.8–1.8 min for type G toxin. Less time was required in phosphate buffer.