A note on the microbial spoilage of undercooked chub-packed luncheon meat

Contrary to expectations slight undercooking (68.5°C instead of 70°C for 90 min) dramatically increased the shelf-life of chub-packed luncheon meat stored at 25°C. The pH of undercooked chubs fell rapidly to below 5.0 as a result of the growth of enterococci. The accumulated acid prevented the growth of Bacillus spores and gave the luncheon meat a not unpleasant tangy flavour. Degradative changes associated with the spoilage of commercially cooked chub-packed luncheon meat did not occur, even after 42 d storage. Apparently, post-cooking fermentation by enterococci can effectively convert a perishable product into a 'shelf stable' one by lowering the pH below 5.0.     

The role of oxygen in the microbial spoilage of luncheon meat cooked in a plastic casing

The shelf life at 25°C of chub-packed luncheon meat was inversely related to oxygen (O₂) availability within the casing. With fibrous casings that are freely permeable to O₂, shelf life was less than 3 d. With plastic casings of low O₂ permeability, the shelf life was 7 d when air was trapped in the emulsion during the casing filling process, 14 d when air was not trapped (normal vacuum-stuffing) and greater than 28 d when vacuum-stuffed chubs were stored in hydrogen (H₂). The initial spoilage bacteria, Bacillus spp., grew only at the surface unless air was trapped in the emulsion when growth occurred throughout the luncheon meat. Bacillus spp. failed to grow on luncheon meat stored under H₂. Oxygen availability within the casing determined both the site and rate of microbial spoilage of chub-packed luncheon meat.     

Heat injury and recovery of Streptococcus faecium associated with the souring of chub-packed luncheon meat

The presence of NaCl in the heating medium provided some protection from lethal heat damage for cells of a Streptococcus faecium strain isolated from luncheon meat whereas the presence of NaNO₂ either alone or in addition to NaCl, had no significant effect on cell survival. Subsequent recovery and growth of heat-damaged cells was retarded by the presence of NaCl. When NaNO₂ was present in addition to NaCl the inhibitory effect of the latter was reduced. These principal components of the luncheon-meat-cure are apparently opposed in their activities on post-heating recovery and growth of Strep. faecium . Product stability, i.e. duration of the lag before growth occurs, is directly related to the severity of the heat treatment and to the concentration of NaCl in the product. Therefore the resistance of pasteurized chub-packed luncheon meat to streptococcal spoilage during storage at temperatures conducive to microbial growth results from a prolonged heat-induced salt-maintained pre-growth adjustment phase rather than to any inherent inhibitory property of the luncheon meat to the growth of non-heat-damaged Strep. faecium cells.     

Heat injury and recovery of Streptococcus faecium associated with the souring of chub-packed luncheon meat

The presence of NaCl in the heating medium provided some protection from lethal heat damage for cells of a Streptococcus faecium strain isolated from luncheon meat whereas the presence of NaNO2 either alone or in addition to NaCl, had no significant effect on cell survival. Subsequent recovery and growth of heat-damaged cells was retarded by the presence of NaCl. When NaNO2 was present in addition to NaCl the inhibitory effect of the latter was reduced. These principal components of the luncheon-meat-cure are apparently opposed in their activities on post-heating recovery and growth of Strep. faecium. Product stability, i.e. duration of the lag before growth occurs, is directly related to the severity of the heat treatment and to the concentration of NaCl in the product. Therefore the resistance of pasteurized chub-packed luncheon meat to streptococcal spoilage during storage at temperatures conducive to microbial growth results from a prolonged heat-induced salt-maintained pre-growth adjustment phase rather than to any inherent inhibitory property of the luncheon meat to the growth of non-heat-damaged Strep. faecium cells.     

Nitrite loss and spoilage microflora development in chub-packed luncheon meat

Residual nitrite was lost from chub-packed luncheon meat during storage through both chemical breakdown and microbial consumption. The relative importance of these mechanisms in this pasteurized product was determined by the speed of development of the spoilage microflora, which is influenced by storage conditions. The nitrite half-life due to chemical loss was 13 d at 25°C and 36 d at 10°C. When microbial growth occurred these half-lives were reduced to 2.6 d and 21 d, respectively. Qualitative differences in the microflora that developed at these two temperatures (denitrifying Bacillus spp. at 25°C and non-denitrifying Streptococcus spp. at 10°C) account for the large temperature effect. Growth of Streptococcus spp. increased the rate of chemical nitrite loss in chubs by reducing the pH value. Nitrite did not inhibit the aerobic growth of either Bacillus or Streptococcus species associated with spoilage but did inhibit the anaerobic growth of Bacillus spp. This bacteriostatic effect of residual nitrite in anaerobic conditions will decrease during storage as nitrite level falls and oxygen penetrates the chub pack. Nitrite-mediated bacteriostasis does not obviate the need for refrigerated storage but does afford a real, if ephemeral, safeguard against spoilage occurring during short periods of temperature abuse.     

Examination of bovine faeces for the isolation and identification of Clostridium species

Freshly cooked luncheon meat in a plastic (PVDC) casing had an aerobic plate count of about 10²/g. The flora was composed of approximately equal numbers of Bacillus and Micrococcus spp. Storage at 10°C for 42 d produced little increase in bacterial numbers, or changes in pH value or glucose content at either the surface or core of the luncheon meat. Storage at 25°C allowed Bacillus spp. to proliferate at the surface. The inhibitory effect of salt and nitrite on the growth of heated Bacillus spores at low redox potentials probably accounts for the absence of growth within the product. Growth at the surface was accompanied by a fall in pH (6.8 to 6.2) and an increase in glucose (1.6 to 3.6 mg/g) and L(+)-lactic acid (1.2 to 2.3 mg/g). By day 14 the Bacillus spp. had been displaced by a Streptococcus sp. (10⁷/g) which remained the dominant organism until the experiment ended on day 28. The pH continued to fall from 5.7 on day 14 to 5.2 on day 28, the L(+)-lactic acid rose to 6.1 mg/g, but the glucose remained constant at the day 7 level (3.6 mg/g). This indicates that glucose converted to lactic acid was largely replaced by hydrolysis of the starch portion of the luncheon meat mediated by amylases produced by the Bacillus microflora. It appears that growth of the Streptococcus is dependent upon the denitrifying activities of the initial Bacillus flora reducing the concentration of nitrite ion to non-inhibitory levels.     

Interactive effects of pH and temperature on the bacteriocin stability by response surface analysis

The combined influence of pH and temperature on bacteriocins produced by three lactic acid bacteria, Pediococcus pentosaceus MMZ26, Enterococcus faecium MMZ17 and Lactococcus lactis MMZ25, isolated from Tunisian traditional dry fermented meat was studied using a second order orthogonal factorial design and response-surface methodology (RSM). This method allows estimating the interactive effects of pH and temperature on the stability of each bacteriocin. The high heat stability of the three bacteriocins was demonstrated, with optimum values at light acidic pH around 5.0, temperature below 90 degrees C and short incubation times. This study contributes to a better understanding of relation between bacteriocins production and stability in order to enhance their, in situ, application as a food and feed biopreservative in fermented and/or heated food products.     

产β—葡聚糖酶菌种T199的选育及发酵条件

大麦为啤酒酿造原料 ,含有由葡萄糖残基通过β 1 ,3 和 1 ,4 糖甙键连接而成的β 葡聚糖。在麦芽汁制备过程中 ,热不稳定的大麦葡聚糖酶不能充分降解β 葡聚糖 ,残留的 β 葡聚糖不仅影响麦芽汁分离和啤酒过滤 ,而且将成为成品啤酒出现混浊和沉淀的因素之一。微生物 β 葡聚糖酶能改善啤酒加工工艺和提高产品质量[1,2 ] 。谷类饲料含有不同于纤维素的 β 葡聚糖[2 ] ,作为抗营养因子 ,β 葡聚糖使饲料具有粘性 ,不能很好的消化利用。β 葡聚糖酶作为饲料添加剂加入到饲料中 ,可以将 β 葡聚糖降解 ,从而提高饲料利用率 ,改善营养吸收。相关…     

Survival or growth of Escherichia coli O157:H7 in a model system of fresh meat decontamination runoff waste fluids and its resistance to subsequent lactic acid stress

A potential may exist for survival of and resistance development by Escherichia coli O157:H7 in environmental niches of meat plants applying carcass decontamination interventions. This study evaluated (i) survival or growth of acid-adapted and nonadapted E. coli O157:H7 strain ATCC 43895 in acetic acid (pH 3.6 +/- 0.1) or in water (pH 7.2 +/- 0.2) fresh beef decontamination runoff fluids (washings) stored at 4, 10, 15, or 25 degrees C and (ii) resistance of cells recovered from the washings after 2 or 7 days of storage to a subsequent lactic acid (pH 3.5) stress. Corresponding cultures in sterile saline or in heat-sterilized water washings were used as controls. In acetic acid washings, acid-adapted cultures survived better than nonadapted cultures, with survival being greatest at 4 degrees C and lowest at 25 degrees C. The pathogen survived without growth in water washings at 4 and 10 degrees C, while it grew by 0.8 to 2.7 log cycles at 15 and 25 degrees C, and more in the absence of natural flora. E. coli O157:H7 cells habituated without growth in water washings at 4 or 10 degrees C were the most sensitive to pH 3.5, while cells grown in water washings at 15 or 25 degrees C were relatively the most resistant, irrespective of previous acid adaptation. Resistance to pH 3.5 of E. coli O157:H7 cells habituated in acetic acid washings for 7 days increased in the order 15 degrees C > 10 degrees C > 4 degrees C, while at 25 degrees C cells died off. These results indicate that growth inhibition by storage at low temperatures may be more important than competition by natural flora in inducing acid sensitization of E. coli O157:H7 in fresh meat environments. At ambient temperatures in meat plants, E. coli O157:H7 may grow to restore acid resistance, unless acid interventions are applied to inhibit growth and minimize survival of the pathogen. Acid-habituated E. coli O157:H7 at 10 to 15 degrees C may maintain a higher acid resistance than when acid habituated at 4 degrees C. These responses should be evaluated with fresh meat and may be useful for the optimization of decontamination programs and postdecontamination conditions of meat handling.     

Development of a Selective Enterococcus Medium Based on Manganese Ion Deficiency, Sodium Azide, and Alkaline pH

Rogosa broth, without its salt supplement and dissolved in deionized water, was adapted for the selective isolation and enumeration of enterococci. This medium supported good growth of enterococci, but it suppressed growth of other lactic acid bacteria. The sensitivity and specificity of the medium were tested after addition of various increasing concentrations of NaN(3) against known strains of enterococci and other bacteria. Many strains of Streptococcus faecium showed low azide tolerance; optimal growth was obtained at a concentration of 0.01% NaN(3), which totally or partially inhibited unrelated species of lactic acid bacteria. The selectivity of the medium was further increased by pH adjustment to 9.6. Carbonate and Tween 80 were added to overcome partial inhibition of enterococcal growth by the new combination of selective conditions. The final medium was evaluated in agar form in isolations from human and animal feces, polluted water, meat, and dairy products. Counts were obtained after 16 to 17 h of incubation at 37 C. The isolates satisfactorily conformed to the group characteristics of enterococci.     

Improvement of Raw Sausage Fermentation by Stress-Conditioning of the Starter Organism Lactobacillus sakei

Effective growth and high acidification activity during meat fermentation are key characteristics of starter lactobacilli to ensure hygienic safety and sensory quality of the product. In this study, we demonstrated that the performance of Lactobacillus sakei in sausage fermentation can be improved by preinoculation treatments with sublethal heat, cold, and salt stress. Sausages were produced and inoculated with stress-treated cells of L. sakei 23 K (pLPV111) and the isogenic mutant of the class III heat-shock repressor CtsR, which was previously shown to exhibit improved growth in fermenting sausages. The pH values of sausages fermented with stressed cells attained defined threshold values in a distinctly shorter time than those inoculated with unstressed cells. In particular, the cold-stressed cells (4 degrees C) reduced the pH to 5.0 within approximately 40 hours compared with approximately 70 hours for untreated cells. This enhanced acidification activity of the cold-stressed cells was consistent with an increased growth rate. Growth studies in culture medium showed that stress-treated cells with improved performance did not exhibit this advantage when exposed to curing salt, one of the major stressors at the beginning of sausage fermentation. Preinoculation stress treatment is a promising way to improve the effectiveness of meat starter lactobacilli.     

Dependence of Clostridium botulinum gas and protease production on culture conditions

Reports that Clostridium botulinum toxin can sometimes be detected in the absence of indicators of overt spoilage led to a systematic study of this phenomenon in a model system. Media with various combinations of pH (5.0 to 7.0) and glucose (0.0 to 1.0%) were inoculated with vegetative cells of C. botulinum 62A and incubated anaerobically at 35 degrees C. Although growth and toxin production occurred at all pH and glucose combinations, accumulation of gas was delayed or absent in media with low pH, low glucose levels, or both. Other proteolytic C. botulinum strains gave similar results. Trypsin activation was required to detect toxin in some low pH cultures. The trypsinization requirement correlated with low proteolytic activity in the cultures. Proteolytic activity of the strains examined was 5- to 500-fold lower in botulinal assay medium than in cooked meat medium. The results indicate that the absence of gas accumulation does not preclude the presence of botulinal toxin and that proteolytic cultures grown under adverse conditions may require trypsinization for the detection of toxin.     

Dependence of Clostridium botulinum gas and protease production on culture conditions.

Reports that Clostridium botulinum toxin can sometimes be detected in the absence of indicators of overt spoilage led to a systematic study of this phenomenon in a model system. Media with various combinations of pH (5.0 to 7.0) and glucose (0.0 to 1.0%) were inoculated with vegetative cells of C. botulinum 62A and incubated anaerobically at 35 degrees C. Although growth and toxin production occurred at all pH and glucose combinations, accumulation of gas was delayed or absent in media with low pH, low glucose levels, or both. Other proteolytic C. botulinum strains gave similar results. Trypsin activation was required to detect toxin in some low pH cultures. The trypsinization requirement correlated with low proteolytic activity in the cultures. Proteolytic activity of the strains examined was 5- to 500-fold lower in botulinal assay medium than in cooked meat medium. The results indicate that the absence of gas accumulation does not preclude the presence of botulinal toxin and that proteolytic cultures grown under adverse conditions may require trypsinization for the detection of toxin.     

Isolation and characteristics of trypsin from pyloric ceca of the starfish Asterina pectinifera

Trypsin was purified from pyloric ceca of the starfish Asterina Pectinifera by ammonium sulfate precipitation, gel filtration, and cation-exchange chromatography. Final enzyme preparation was nearly homogeneous in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its molecular weight was estimated as approximately 28000. Optimum pH and temperature of A. pectinifera trypsin for hydrolysis of N(alpha)-p-Tosyl-L-arginine methyl ester hydrochloride were approximately pH 8.0 and 55 degrees C, respectively. A. pectinifera trypsin was unstable at above 50 degrees C and below pH 5.0, and was not activated by adding Ca(2+). The N-terminal amino acid sequence of A. pectinifera trypsin, IVGGHEF, was found.     

Characterization of a thermostable endo-beta-1,4-D-galactanase from the hyperthermophile Thermotoga maritima

A putative endo-beta-1,4-D-galactanase gene of Thermotoga maritima was cloned and overexpressed in Escherichia coli. The recombinant enzyme hydrolyzed pectic galactans and produced D-galactose, beta-1,4-D-galactobiose, beta-1,4-D-galactotriose, and beta-1,4-D-galactotetraose. The enzyme displayed optimum activity at 90 degrees C and pH 7.0. It was slowly inactivated above pH 8.0 and below pH 5.0 and stable at temperatures up to 80 degrees C.     

Influence of the natural microbial flora on the acid tolerance response of Listeria monocytogenes in a model system of fresh meat decontamination fluids

Depending on its composition and metabolic activity, the natural flora that may be established in a meat plant environment can affect the survival, growth, and acid tolerance response (ATR) of bacterial pathogens present in the same niche. To investigate this hypothesis, changes in populations and ATR of inoculated (10(5) CFU/ml) Listeria monocytogenes were evaluated at 35 degrees C in water (10 or 85 degrees C) or acidic (2% lactic or acetic acid) washings of beef with or without prior filter sterilization. The model experiments were performed at 35 degrees C rather than lower (8.0 log CFU/ml) by day 1. The pH of inoculated water washings decreased or increased depending on absence or presence of natural flora, respectively. These microbial and pH changes modulated the ATR of L. monocytogenes at 35 degrees C. In filter-sterilized water washings, inoculated L. monocytogenes increased its ATR by at least 1.0 log CFU/ml from days 1 to 8, while in unfiltered water washings the pathogen was acid tolerant at day 1 (0.3 to 1.4 log CFU/ml reduction) and became acid sensitive (3.0 to >5.0 log CFU/ml reduction) at day 8. These results suggest that the predominant gram-negative flora of an aerobic fresh meat plant environment may sensitize bacterial pathogens to acid.     

Purification and characterization of two proteolytic enzymes in the cotyledons of germinating lentils

Two proteases, one peptidehydrolase and one aminopeptidase, have been purified to homogeneity from cotyledons of germinating seeds of Lens culinaris Med. Peptidehydrolase has an apparent molecular weight of 89,000 and an isoelectric point of 4.7. Peptidehydrolase activity was not affected by metal chelators but it was affected by N-bromosuccinimide, phenylmethylsulfonyl fluoride and N-ethylmaleimide, suggesting the presence of tryptophan and serine residues together with free--SH groups in its active site. Peptidehydrolase activity was maximally active from pH 6.0 to 9.0 being practically zero below pH 5.0. It was stable at temperatures up to 40 degrees C, and complete inactivation was obtained at or over 70 degrees C. Aminopeptidase has an apparent molecular weight of 83,000 and an isoelectric point of 4.5. Its activity was affected by N-bromosuccinimide, suggesting the presence of tryptophan residues in its active site. The aminopeptidase presents its maximal activity at pH 5.5. It was stable at temperatures up to 40 degrees C, and complete inactivation was detected at over 70 degrees C.     

The Growth of Microbacterium thermosphactum on Beef

Microbacterium thermosphactum grew aerobically at 5°C on beef when the pH was as low as 5.4. It did not grow anaerobically if the pH of the meat was below 5.8, but grew readily at pH 6.0 or higher. On vacuum-packaged beef of pH 5.8 or lower, growth was dependent not only on pH but also on the oxygen permeability of the packaging film. Above pH 5.9 growth occurred with all the films tested.     

Production and characterization of tannase from Bacillus cereus KBR9

A tannase-producing soil bacteria has been isolated and identified as Bacillus cereus. It can degrade tannic acid and produce maximum tannase (0.22 U/ml) at stationary phases of growth (24 h). Maximum growth and enzyme production occurred with initial medium pH of 4.5-5.0. Partial purified tannase showed optimum activity at pH 4.5 and 40 degrees C. It remains stable up to 30 degrees C and pH 4.5 to 5.0. The enzyme is salt tolerant, stable up to 2 m of NaCl and retains 82% original activity in 3 m.     

枯草芽孢杆菌中性β—甘露聚糖酶的产生及性质

由土壤中分离出一株产中性β甘露聚糖酶的枯草芽孢杆菌（Ｂａｃｉｌｕｓｓｕｂｔｉｌｉｓ）,编号ＢＭ９６０２。该菌在液体培养条件下,产生中性β甘露聚糖酶。多糖能作为碳源,而单糖不能作为碳源;有机氮源优于无机氮源。产酶最适培养基组成：魔芋粉４％,牛肉蛋白胨和酵母膏各１％。产酶最适培养条件：培养基起始ｐＨ８５,３５℃,振荡培养３６ｈ。以槐豆胶为底物,培养滤液中性β甘露聚糖酶活力为９６ＩＵ／ｍＬ。酶在ｐＨ５０～１００和５０℃下稳定;作用最适条件为ｐＨ６０和５０℃;水解魔芋粉和槐豆胶均产生寡聚糖。