Storage of poultry meat under modified atmospheres or vacuum packs: possible role of microbial metabolites as indicator of spoilage

The effect of carbon dioxide (100%), nitrogen (100%), carbon dioxide/oxygen (20% : 80%) or vacuum pack at 3 and 10°C was studied on the microbial flora, in skinless poultry breast fillets or thigh meat. Lactic acid bacteria and Brochothrix thermosphacta were the predominant organisms in samples stored in vacuum packs, carbon dioxide and nitrogen. Pseudomonads grew only in oxygen/carbon dioxide packaging systems. The concentration of lactate diminished in both thigh and breast meat during storage at 3 and 10°C. This decrease was more pronounced in thigh meat stored under 20% : 80% carbon dioxide/oxygen. Acetate increased to varying degrees in all samples regardless of the storage conditions.     

The characteristics of a new non-spore-forming cellulolytic mesophilic anaerobe strain CM126 isolated from municipal sewage sludge

A new mesophilic anaerobic cellulolytic bacterium, CM126, was isolated from an anaerobic sewage sludge digester. The organism was non-spore-forming, rod-shaped, Gram-negative and motile with peritrichous flagella. It fermented microcrystalline Avicel cellulose, xylan, Solka floc cellulose, filter paper, L-arabinose, D-xylose, β-methyl xyloside, D-glucose, cellobiose and xylitol and produced indole. The % G + C content was 36. Acetic acid, ethanol, lactic acid, pyruvic acid, carbon dioxide and hydrogen were produced as metabolic products. This strain could grow at 20–44·5°C and at pH values 5·2–7·4 with optimal growth at 37–41·5°C and pH 7. Both endoglucanase and xylanase were detected in the supernatant fluid of a culture grown on medium containing Avicel cellulose and cellobiose. Exoglucanase could not be found in either supernatant fluid or the cell lysate. When cellulose and cellobiose fermentation were compared, the enzyme production rate in cellobiose fermentation was higher than in cellulose fermentation. The optimum pH for both enzyme activities was 5·0, the optimum temperature was 40°C for the endoglucanase and 50°C for the xylanase. Both enzyme activities were inhibited at 70°C. Co-culture of this organism with a Methanosarcina sp. (A145) had no effect on cellulose degradation and both endoglucanase and xylanase were stable in the co-culture.     

Incidence and Spoilage Potential of Isolates from Vacuum-packaged Meat of High pH Value

Meat of high pH value (6·6) showing dark-cutting characteristics was vacuum-packaged and stored for up to 8 weeks at 0–2°C. 'Off'-odours were detected on opening the packages after 6 weeks of storage. Total counts at this stage were ca. 10⁷/cm² of which lactobacilli were the major component, with ca. 10⁶/cm² Gram negative organisms. Psychrotrophic Enterobacteriaceae represented a major proportion of the microflora only after the full 8 weeks of storage and were not detected previously. Aerobic storage of steaks cut from the vacuum packaged meat stored for 8 weeks resulted in a predominantly Gram negative spoilage flora.
Inoculation studies on meat of normal pH value (5·4) and appearance using representative isolates from the vacuum-packaged meat microflora indicated that most of the test organisms were capable of causing spoilage under aerobic conditions but few under vacuum-packaging when incubated at 4°C. On meat of higher pH value (6·15) many of the Gram negative isolates did not grow as well, whereas the Gram positive isolates grew better than on meat of normal pH value when held under aerobic conditions. Under vacuum-packaging all but one isolate grew as well or better on meat of high pH value than on normal meat at 4°C and objectionable odours were more marked.     

The Effects of Sulphite Preservative in British Fresh Sausages

S ummary . The bacteriology of pork and beef sausages, with and without metabisulphite preservative, when stored at room temperature and 3—5° is described. The preservative has a marked effect on microbial growth in sausages stored at room temperature, particularly with regard to the organisms found on plates incubated at 37°. Of the latter, coli-aerogenes and other Gram negative organisms were the most inhibited by the preservative.     

Effect of Packaging under Carbon Dioxide, Nitrogen or Air on the Microbial Flora of Pork Stored at 4°C

Changes in the microbial flora of pork packed in laminated plastic bags and stored at 4 °C were studied in an initial atmosphere of carbon dioxide, nitrogen or air. The time needed for the total aerobic count at 28 °C to reach 5 × 10⁶ organisms/cm² was about 7 times longer in carbon dioxide than in air, whilst in nitrogen it was about twice as long.
The predominant organisms on fresh pork taken directly from the processing line were: Acinetobacter calcoaceticus , non-fluorescent Pseudomonas spp. and Flavobacterium spp. After storage in air for 7 d, more than 90% of the flora consisted of non-fluorescent Pseudomonas spp. After storage in nitrogen for 10 d, 70% of the flora consisted of non-fluorescent Pseudomonas spp. with lower levels of fluorescent Pseudomonas spp., Kurthia zopfii, Aeromonas hydrophila and Lactobacillus plantarum. The non-fluorescent Pseudomonas spp. could be divided into three different groups, on proteolytic and lipolytic ability; the distribution of the groups was markedly different between pork loins stored in air and nitrogen.
On pork stored in carbon dioxide for 21 d the flora consisted of L. plantarum together with lower levels of heterofermentative lactic acid bacteria. When the storage time in carbon doxide was prolonged to 35 d, the proportion of heterofermentative lactic acid bacteria increased to about 50% of the flora.     

Significance of temperature and preincubation temperature on survival of Listeriamonocytogenes at pH 4·8

Listeria monocytogenes is a food-borne pathogenic bacterium that can be found in softcheese. At the beginning of cheese ripening, the pH is about 4·85–4·90. The aimof this work was to study the influence of temperature, preincubation temperature (temperature atwhich the inoculum was cultivated) and initial bacterial concentration on the survival of L.monocytogenes (strain Scott A) at pH 4·8. It was demonstrated in an earlier study thatthese factors did influence growth kinetics. Survival studies of L. monocytogenes weredone in a laboratory broth simulating cheese composition. Four test temperatures (2, 6, 10 and14°C) and two preincubation temperatures were studied (30°C or the test temperature). Listeria monocytogenes (strain Scott A) was unable to grow at pH 4·8 under allconditions tested. The time for 10% survival was about 11 and 2 d, at 2°C with preincubationat 2°C and 30°C, respectively; 9 d at 6°C with preincubation at 6°C; 4 d at 6°Cwith preincubation at 30°C; and 1 d at 14°C with preincubation at 14°C or at 30°C.The results show that survival of L. monocytogenes (strain Scott A) at pH 4·8 is notdependent on initial bacterial concentration but on both the test and preincubation temperatures.     

Survival and growth of Cronobacter species (Enterobacter sakazakii) in wheat-based infant follow-on formulas

Aims:  To determine the survival and growth characteristics of Cronobacter species ( Enterobacter sakazakii ) in infant wheat-based formulas reconstituted with water, milk, grape juice or apple juice during storage.
Methods and Results:  Infant wheat-based formulas were reconstituted with water, ultra high temperature milk, pasteurized grape or apple juices. The reconstituted formulas were inoculated with Cronobacter sakazakii and Cronobacter muytjensii and stored at 4, 25 or 37°C for up to 24 h. At 25 and 37°C, Cronobacter grew more (>5 log₁₀) in formulas reconstituted with water or milk than those prepared with grape or apple juices ( c. 2–3 log₁₀). The organism persisted, but did not grow in any formulas stored at 4°C. Formulas reconstituted with water and milk decreased from pH 6·0 to 4·8–5·0 after 24 h, whereas the pH of the formulas reconstituted with fruit juices remained at their initial pH values, c. pH 4·8–5·0.
Conclusions:  Cronobacter sakazakii and C. muytjensii can grow in reconstituted wheat-based formulas. If not immediately consumed, these formulas should be stored at refrigeration temperatures to reduce the risk of infant infection.
Significance and Impact of the Study:  The results of this study will be of use to regulatory agencies and infant formula producers to recommend storage conditions that reduce the growth of Cronobacter in infant wheat-based formulas.     

Production of cell wall dissolving enzymes by Erwinia carotovora subsp. atroseptica in vitro at 27°C and 30°5C

Cultures of Erwinia carotovora subsp. atroseptica , grown at 27°C and 30·°C in different liquid media were assayed for activities of pectate lyase, polygalacturonase and cellulase. Total production of both pectate lyase and of polygalacturonase was 3–6 times less at 30·5°C than at 27°C; secretion of pectate lyase was similarly affected. Cellulase was cell bound and its production was not affected by the temperatures investigated. Growth, protein synthesis and protease activity were similar at the two temperatures and production of enzyme activity at 27°C and 30·5°C was independent of the growth medium.     

The combined effect of sub-optimal temperature and sub-optimal pH on growth and toxin formation from spores of Clostridium botulinum

Low-acid foods (pH ≥ 4.5) are not sufficiently acidic to prevent growth of Clostridium botulinum in otherwise optimal conditions. The combination of sub-optimal pH and sub-optimal temperature may, however, result in a very significant reduction in the risk of growth of this bacterium compared with the risk in optimal conditions. The combined effect of incubation temperatures of 12° and 16°C and pH values between 5·2 and 5·5 on growth and toxin production from spores of Cl. botulinum during incubation for 28 d has been investigated. Growth and formation of toxin (type B) were detected only in medium at pH 5·5 and incubated at 16°C, corresponding to a probability of growth from a single spore within 14 d of 1·6 × 10^-5. The probability of growth in 28 d in the remaining conditions was <9 × 10^-6. After transfer of inoculated media from 12° to 30°C growth occurred at pH 5·2–5·5 within 19 d. After transfer of inoculated media from 12° to 20°C growth occurred at pH 5·5 and 5·4 but not at pH 5·3 or 5·2 in 40 d. Growth at pH 5·2–5·5 was accompanied by formation of toxin, in most cases of types A or B. In addition to the effect of sub-optimal temperature and pH, chelation of divalent metal ions by citrate may have contributed to inhibition.     

Effect of salt concentration and temperature on survival of Legionella pneumophila

The effects of various concentrations of sodium chloride solutions (0·1%–3%) and different temperatures (4, 10, 20, 30 and 37 °C) on survival of Legionella pneumophila were investigated. It was found that at temperatures between 4 °C and 20 °C, Legionella organisms survived in salt solutions up to 3% NaCl. Only the combination of high temperatures, i. e. 30 °C and 37 °C, with NaCl concentrations over 1·5%, reduced cell numbers significantly. It was interesting to note that the addition of small amounts of NaCl (0·1%–0·5%) enhanced survival of Leg. pneumophila , suggesting a protective effect of NaCl. In order to obtain information about conditions encountered in the environment, the survival experiments were repeated in sterile sea water from the Baltic Sea and the North Sea. The marked bacterial die-off, especially at higher temperatures, was not observed in natural sea water. All these results indicate that Leg. pneumophila can survive in the marine environment.     

The effects of temperature on muscle pH, adenylate and phosphogen concentrations in Oreochromis alcalicus grahami, a fish adapted to an alkaline hot-spring

The concentrations of phosphorylcreatine (PCr), adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), inorganic phosphate (Pi), pyruvate and lactate were determined in freeze-clamped fast muscle samples from Oreochromis alcalicus grahami a fish adapted to extreme alkalinity (∼ pH 10·0) and high temperatures (Lake Magadi, Kenya). Specimens were analysed from both geothermally heated hotsprings (35–37°C) and from isolated cool pools (28°C) and from stocks acclimated to 20°C in the laboratory. The ratios of (ATP)/(ADP) and (ATP)/(ADP) (Pi) decreased with increasing body temperature consistent with an increase in glycolysis and tissue respiration rates, respectively. The apparent equilibrium constant of creatine kinase (K_CK), (creatine) (ATP)/(phosphorylcreatine) (ADP) was found to decrease with increasing temperature: 20·2 (20°C), 13·9 (28°C), 8·0 (37°C). A near constant muscle and blood pH (or slight increase in alkalinity with higher temperatures) was found regardless of body temperature (Blood pH 7·64, 7·74, muscle pH 7·27, 7·51 at 20°C and 35°C, respectively). These results are consistent with an unusual pattern of acid-base regulation in this species.     

Growth and Sporulation of Clostridium welchii in Breast and Leg Muscle of Poultry

Growth of a heat resistant, food poisoning strain of Clostridium welchii was followed in raw, minced breast and leg muscle of the chicken. Within the range 22–50° growth was slightly more rapid in the leg (pH 6·5–6·7) than in the breast (pH 5·6–5·7) and was fastest in leg muscle at 50°. No growth occurred at 15 or 52°.
In a comparison between chicken and turkey, inoculated breast and leg muscle were cooked for 1 h at 85° and held at 37°. Multiplication of surviving organisms was initiated much more rapidly in chicken than in turkey meat, though the growth rates were comparable in each case.
Sporulation of several strains of CI. welchii , including other heat resistant, food poisoning types, was generally 10–100 times greater in leg than in breast muscle of the chicken. Differences in sporulation could be attributed both to differences in pH and type of meat.     

Effects of essential oil from mint (Mentha piperita) on Salmonella enteritidis and Listeria monocytogenes in model food systems at 4° and 10°C

The effect of mint ( Mentha piperita ) essential oil (0·5, 1·0, 1·5 and 2·0%, v/w) on Salmonella enteritidis and Listeria monocytogenes in a culture medium and three model foods; tzatziki (pH 4·5), taramosalata (pH 5·0) and pâté (pH 6·8), inoculated at 10⁷ cfu g^-1, at 4° and 10°C for ca 1 week was studied. In the culture medium supplemented with the essential oil, no growth was observed over 2 d at 30°C determined by a conductance method with a Malthus 2000 growth analyser. Salmonella enteritidis died in tzatziki in all treatments and declined in the other foods except for pâté at 10°C as judged with viable counts. Listeria monocytogenes populations showed a declining trend towards the end of the storage period but was increased in pâté. Mint essential oil antibacterial action depended mainly on its concentration, food pH, composition, storage temperature and the nature of the micro-organism.     

Tannin impacts on microbial diversity and the functioning of alpine soils: a multidisciplinary approach

In alpine ecosystems, tannin-rich-litter decomposition occurs mainly under snow. With global change, variations in snowfall might affect soil temperature and microbial diversity with biogeochemical consequences on ecosystem processes. However, the relationships linking soil temperature and tannin degradation with soil microorganisms and nutrients fluxes remain poorly understood. Here, we combined biogeochemical and molecular profiling approaches to monitor tannin degradation, nutrient cycling and microbial communities (Bacteria, Crenarcheotes, Fungi) in undisturbed wintertime soil cores exposed to low temperature (0°C/−6°C), amended or not with tannins, extracted from Dryas octopetala . No toxic effect of tannins on microbial populations was found, indicating that they withstand phenolics from alpine vegetation litter. Additionally at −6°C, higher carbon mineralization, higher protocatechuic acid concentration (intermediary metabolite of tannin catabolism), and changes in fungal phylogenetic composition showed that freezing temperatures may select fungi able to degrade D. octopetala 's tannins. In contrast, negative net nitrogen mineralization rates were observed at −6°C possibly due to a more efficient N immobilization by tannins than N production by microbial activities, and suggesting a decoupling between C and N mineralization. Our results confirmed tannins and soil temperatures as relevant controls of microbial catabolism which are crucial for alpine ecosystems functioning and carbon storage.     

Effects of environmental factors on survival, growth, and production of American shad larvae

Episodic increases in temperature of 5°C above 20° C, over 48 h or declines in pH of 1·0 unit from pH 7·0 reduced survival of yolk-sac and feeding-stage larvae of American shad Alosa sapidissima . Over 16 days all measures of survival, growth, and production were more favourable at each higher temperature in the 15–25° C range. More favourable responses were also obtained at the higher prey level (500 v . 50 Artemia nauplii l^-1) and at the higher pH (7·5 v . 6·5). Combinations of high temperature and high prey levels, at pH 7·5, led to highest larval production. Little growth or production occurred at 15° C, regardless of pH or prey level. The effect of pH was strong with respect to survival, but weak with respect to growth. In attempts to restore American shad populations by larval stocking, release times and sites can be critical to optimize survival and eventual returns. Releases of larvae potentially will be most effective when made at temperatures >20° C, pH>7·0, and prey levels >50 1^-1. These conditions are most likely to occur in Maryland tributaries of Chesapeake Bay between mid-May and early June.     

Isolation of Yersinia enterocolitica-resembling Organisms and Alteromonas putrefaciens from Vacuum-Packed Chilled Beef Cuts

Yersinia enterocolitica -resembling organisms were found at levels of 10⁷/g on a high pH (pH ≧ 6·0) vacuum-packaged beef striploin held for 6 weeks at 0·2°C, but did not exceed 10⁵/g on normal pH (pH < 6·0) striploins held for 10 weeks. Gram negative bacteria that produced H₂S on peptone iron agar were isolated from high pH vacuum packed striploins. These organisms were identified as Alteromonas putrefaciens . They attained levels of about 10⁷/g in 6 weeks at 0–2°C, at which time greening of the fat surface and 'drip'had occurred. On meat of normal pH, counts of A. putrefaciens were less than 10⁴/g after 6 weeks and no greening was evident.     

Scaling of enzyme activity in larval herring and plaice: effects of temperature and individual growth rate on aerobic and anaerobic capacity

Herring Clupea harengus and plaice Pleuronectes platessa were reared at 8 and 12° C from the fertilized egg to a larval age of up to 600 degree-days. Soluble protein as well as the activities of both citrate synthetase (CS) and lactate dehydrogenase (LDH) were measured in homogenate supernatants of individual larvae at 10° C. Scaling factors were calculated using the expression y = ax^b where y is the enzyme activity, x the protein content of the larva a is a constant and b the scaling factor. All scaling factors showed significant differences between the species. Within species, the scaling factors for CS activity were either small or not significantly different between the two rearing temperatures, but the scaling factors for the LDH activities were significantly different at the two temperatures for both species. Herring larvae, which had higher LDH activities when newly hatched, showed smaller scaling factors for LDH ( b =1·42 at 8°C and b =1·07 at 12° C) than plaice ( b =2·11 at 8°C and b =1·45 at 12° C). Activities converged as the larvae grew. The results of the current study together with reanalysis of data from the literature indicate an increasing aerobic and anaerobic capacity during the larval stage of fishes (positive allometry).     

Habituation to acid in Escherichia coli: conditions for habituation and its effects on plasmid transfer

Induction of acid resistance (habituation) in Escherichia coli at pH 5·0 took ca 5 min in broth at 37°C and 30–60 min in minimal medium. Induction occurred at a range of pH values from 4·0 to 6·0; it was dependent on continuing protein and RNA synthesis but substantial acid resistance appeared in the presence of nalidixic acid. Acid resistance was long-lasting; organisms grown at pH 5·0 retained most of their resistance after 2 h growth at pH 7·0. Organisms grown at pH 5·0 showed increased synthesis of a number of cytoplasmic proteins compared with the level in cells grown at pH 7·0. DNA repair-deficient strains carrying recA, uvrA or polAl mutations were more acid-sensitive than the repair-proficient parents but were able to habituate at pH 5·0. Organisms grown at pH 5·0 transferred the ColV plasmid much more effectively at acid pH than did those grown at pH 7·0 and habituated recipients appeared better able to repair incoming acid-damaged plasmid DNA than did those that were non-habituated. Induction of acid resistance at pH 5·0 may be significant for the survival of organisms exposed to periodic discharges of acid effluent in the aquatic environment and habituation may also allow plasmid transfer and repair of acid-damaged plasmid DNA during or after such exposure.     

Production and partial characterization of lipases from a newly isolated Penicillium sp. using experimental design

Aims:  The objective of this work was to investigate the lipase production by a newly isolated Penicillium sp . , using experimental design technique, in submerged fermentation using a medium based on peptone, yeast extract, NaCl and olive oil, as well as to characterize the crude enzymatic extracts obtained.
Methods and Results:  Lipase activity values of 9·5 U ml⁻¹ in 96 h of fermentation was obtained at the maximized operational conditions of peptone, yeast extract, NaCl and olive oil concentrations (g l⁻¹) of 20·0, 5·0, 5·0 and of 10·0 respectively. The partial characterization of crude enzymatic extract obtained by submerged fermentation showed optimum activity at pH range from 4·9 to 5·5 and temperature from 37°C to 42°C. The crude extract maintained its initial activity at freezing temperatures up to 100 days.
Conclusions:  A newly isolated strain of Penicillium sp . used in this work yielded good lipase activities compared to the literature.
Significance and Impact of the Study:  The growing interest in lipase production is related to the potential biotechnological applications that these enzymes present. New lipase producers are relevant to finding enzymes with different catalytic properties of commercial interest could be obtained, without using genetically modified organisms (GMO).     

Antimicrobial efficacy of potassium salts of four parabens

Summary The antimicrobial activity of the soluble potassium salts of methyl, ethyl, propyl, and butyl parabens were evaluated to determine whether they would be more effective than their respective parabens (esters ofp-hydroxybenzoic acids). The potassium salts of the methyl and ethyl parabens as well as methyl and ethyl parabens were microbiocidal against the fungusAspergillus niger and five bacteria, whereas the potassium salts of propyl and butyl parabens and their respective parabens were not microbiocidal against all the test organisms. In the presence of several ingredients frequently used in pharmaceutical and cosmetic formulations, ethylenediaminetetraacetate (EDTA) and magnesium hydroxide did not interfere with the antimicrobial activity of the potassium salts of parabens and appeared to be microbiocidal against three of four test organisms. Simethicone and Tween 80 interfered with the antimicrobial activity of the preservatives. At pH 4–6, the potassium salt of butyl paraben, the only preservative tested, was active against more organisms than at pH 7–8. Overall, the highly soluble potassium salts of parabens showed microbiocidal activity against more of the test organisms than the less soluble parabens.