Sensitivity of bacteria to irradiation on poultry meat under various atmospheres

The sensitivity of seven bacterial species inoculated on to sterile poultry meat and irradiated under various atmospheres (air, CO₂, vacuum and nitrogen) was investigated. The irradiation sensitivities of Streptococcus faecalis and Staphylococcus aureus were unaffected by atmosphere. The other micro-organisms ( Pseudomonas putida, Salmonella typhimurium, Escherichia coli, Moraxella phenylpyruvica and a Lactobacillus sp.) were more sensitive (their D₁₀ values decreased) when irradiated in atmospheres other than air. In general, a vacuum or CO₂ atmosphere during irradiation was found to have the most lethal effect. Plating media were found to have a significant effect on the recovery of irradiated E. coli and Salm. typhirium.     

Growth/survival of Salmonella enteritidis on fresh poultry and fish stored under vacuum or modified atmosphere

G.-J.E. NYCHAS AND C.C. TASSOU. 1996. The effect of vacuum and modified atomosphere packaging on the growth/survival of Samonella entertidis on fresh poultry and fish (Boops boos) is described. Salmonella enteritidis survived but did not grow significantly in all samples (poultry or fish) at 3^oC. At 10^oC the numbers of Salm. enteritidis increased rapidly in vacuum-packed samples and in samples flushed with 100% N₂, 20%, CO²/80% O² of both types of proteinaceous food. Growth was also evident in fish and poultry flushed with 100% CO²; however the rate of growth was greater in fish samples rather than in poultry.     

Microbial interaction in cooked cured meat products under vacuum or modified atmosphere at 4 degrees C

AIMS: To investigate the antagonistic activity of two lactic acid strains against the spoilage microflora in cooked cured meat products, vacuum or modified atmosphere packed at 4 degrees C and to determine the inhibitory capacity of their bacteriocins. METHODS AND RESULTS: Frankfurter-type sausages and sliced cooked cured pork shoulder were inoculated with Leuconostoc mesenteroides L124 and Lactobacillus curvatus L442 or with their bacteriocins. The microbial, physico-chemical (pH, L- and D-lactate, acetate and ammonia) and colour changes were studied. Results under vacuum packaging showed that in the uninoculated samples of the pork product the spoilage microflora grew but in the inoculated ones the spoilage microorganisms (e.g. Brochothrix thermosphacta and enterococci) reduced during the storage. This observation was more pronounced in the samples with the addition of bacteriocins. In the frankfurter-type sausages the spoilage microflora did not grow in the uninoculated and inoculated samples. In the modified atmosphere enriched in CO2 the population of spoilage microflora remained at low levels in both products, indicating that CO2 has an effect on the spoilage microorganisms' growth. In the pork product the concentrations of acetate and d-lactate increased while L-lactate decreased, but in the frankfurter-type sausages increase of acetate and D-lactate was not observed. CONCLUSIONS: Lactic acid strains had an effect on the spoilage microflora growth but did not affect, negatively, the organoleptic properties of the products. These strains may be used as biopreservative cultures or their bacteriocins could be an important contribution to microbiological quality of meat products. SIGNIFICANCE AND IMPACT OF STUDY: Establishment of biopreservation as a method for extension of shelf life of meat products.     

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.     

The role of thiol antioxidants in restoring mitochondrial functions modified by microbial metabolites

The effects of phenolic acids of microbial origin on mitochondrial functions and the possibility of removing their effects by thiol antioxidants dithiotreitol and N-acetylcysteine were studied. The action of some phenolic acids on the redox state of NADH, the membrane potential and calcium capacity of mitochondria is due to their interaction with thiol groups. Partial restoration of mitochondrial functions occurred in the presence of dithiotreitol and N-acetylcysteine, full recovery (short-term duration) was promoted by the combined action of dithiotreitol and menadione (vitamin K₃). It was found that the protective effect of thiol antioxidants became prooxidant if the medium contained free iron and compounds with quinone structure, capable of entering into a redox cycle with thiols. It is shown that the interaction of thiols with iron and menadione is accompanied by absorption of oxygen to form superoxide anion. Prooxidant effect of thiol antioxidants may explain the absence of the protective effect at the later stages of sepsis and systemic inflammatory syndrome.     

Light lamb meat quality packed under modified atmospheres: effect of stunning systems (electrically v. gas)

In the present study, the effect of type of stunning (electrical v. gas) and the gas mixtures in a modified atmosphere (MA) packing system (MA-A: 30% CO2 + 70% O2; MA-B: 30% CO2 + 69.3% N2 + 0.7% CO; MA-C: 40% CO2 + 60% N2) on light lamb meat quality, obtained by 20 males of Manchego Spanish breed, was evaluated at 7, 14 and 21 days post packing. For all ageing times, gas-stunned groups reached the highest pH (P < 0.001) and shear force (SF) values (P < 0.05), and at 21 days, water losses were highest; in contrast, meat colour coordinates were lower (P < 0.001). In general, MA-B showed the highest stability for colour coordinates whereas the lowest tenderness (high SF) was found on MA-A. In this study, the gas mixture with a low carbon monoxide level (MA-B) promoted a higher colour stability and good tenderness, in comparison to the other two blends. In contrast, the conventional packaging system (type A, high oxygen levels) had a reduced tenderness and major colour instability.     

A rapid chemical spot test for the detection of lactic acid as an indicator of microbial spoilage in preserved foods

A ckland , M.R. & R eeder , J.E. 1984. A rapid chemical spot test for the detection of lactic acid as an indicator of microbial spoilage in preserved foods. Journal of Applied Bacteriology 56 , 415–419.
A rapid method for the detection of lactic acid in preserved foods has been developed, based upon a chemical spot test which does not require solvent extraction or derivatisation of the lactic acid in the sample. The test can be completed within 5 min and was shown to confirm microbial spoilage detected by cultural techniques in a range of preserved foods. In addition the test was able to indicate microbial spoilage in samples where cultural techniques failed because the spoilage organisms were dead. The test may not be appropriate for products which have a naturally high lactic acid content.     

A rapid chemical spot test for the detection of lactic acid as an indicator of microbial spoilage in preserved foods

A rapid method for the detection of lactic acid in preserved foods has been developed, based upon a chemical spot test which does not require solvent extraction or derivatisation of the lactic acid in the sample. The test can be completed within 5 min and was shown to confirm microbial spoilage detected by cultural techniques in a range of preserved foods. In addition the test was able to indicate microbial spoilage in samples where cultural techniques failed because the spoilage organisms were dead. The test may not be appropriate for products which have a naturally high lactic acid content.     

Growth of Listeria monocytogenes in meat juice under a modified atmosphere at 4°C with or without members of a microbial association from chilled lamb under a modified atmosphere

Listeria monocytogenes together with Gram-positive and Gram-negative bacteria was grown in axenic cultures in meat juice under an atmosphere enriched with carbon dioxide at 4°C. The size of the final population of listeriae was determined by that of the initial inoculum. Growth of L. monocytogenes ceased with glucose depletion. Glucose-6-phosphate was not catabolized by listeriae. In aerobic axenic cultures generation times of L. monocytogenes were shorter and glucose depletion was faster than with a carbon dioxide-enriched atmosphere. Oxidation of L- and D-lactic to acetic acid was probably associated with the subsequent die-off of L. monocytogenes.     

Development of a microbial model for the combined effect of temperature and pH on spoilage of ground meat, and validation of the model under dynamic temperature conditions

The changes in microbial flora and sensory characteristics of fresh ground meat (beef and pork) with pH values ranging from 5.34 to 6.13 were monitored at different isothermal storage temperatures (0 to 20 degrees C) under aerobic conditions. At all conditions tested, pseudomonads were the predominant bacteria, followed by Brochothrix thermosphacta, while the other members of the microbial association (e.g., lactic acid bacteria and Enterobacteriaceae) remained at lower levels. The results from microbiological and sensory analysis showed that changes in pseudomonad populations followed closely sensory changes during storage and could be used as a good index for spoilage of aerobically stored ground meat. The kinetic parameters (maximum specific growth rate [mu(max)] and the duration of lag phase [lambda]) of the spoilage bacteria were modeled by using a modified Arrhenius equation for the combined effect of temperature and pH. Meat pH affected growth of all spoilage bacteria except that of lactic acid bacteria. The "adaptation work," characterized by the product of mu(max) and lambda(mu(max) x lambda) was found to be unaffected by temperature for all tested bacteria but was affected by pH for pseudomonads and B. thermosphacta. For the latter bacteria, a negative linear correlation between ln(mu(max) x lambda) and meat pH was observed. The developed models were further validated under dynamic temperature conditions using different fluctuating temperatures. Graphical comparison between predicted and observed growth and the examination of the relative errors of predictions showed that the model predicted satisfactorily growth under dynamic conditions. Predicted shelf life based on pseudomonads growth was slightly shorter than shelf life observed by sensory analysis with a mean difference of 13.1%. The present study provides a "ready-to-use," well-validated model for predicting spoilage of aerobically stored ground meat. The use of the model by the meat industry can lead to effective management systems for the optimization of meat quality.     

The microbial association of Greek taverna sausage stored at 4 and 10 degrees C in air, vacuum or 100% carbon dioxide, and its spoilage potential

Strains of the Lactobacillus sakei/curvatus group, mainly non-slime-producing Lact. sakei, dominated the microbial flora of industrially manufactured taverna sausage, a traditional Greek cooked meat, stored at 4 degrees C and 10 degrees C in air, vacuum and 100% CO2. Atypical, arginine-positive and melibiose-negative strains of this group were isolated. The isolation frequency of Lact. sakei/curvatus from sausages stored anaerobically was as high as 92-96%, while other meat spoilage organisms were practically absent. Conversely, in air-stored sausages, leuconostocs, mainly Leuconostoc mesenteroides ssp. mesenteroides, had a considerable presence (14-21%), whereas Brochothrix thermosphacta, pseudomonads and Micrococcaceae grew, but failed to increase above 10(5) cfu g(-1) in all samples during storage. Only yeasts were able to compete against LAB and reached almost 10(7) cfu g(-1) after 30 d of aerobic storage at 10 degrees C. The great dominance (> 10(8) cfu g(-1)) of LAB caused a progressive decrease of pH and an increase of the concentration of L-lactate, D-lactate and acetate in all sausage packs. The growth of LAB and its associated chemical changes were more pronounced at 10 degrees C than 4 degrees C. At both storage temperatures, L-lactate and acetate increased more rapidly and to a higher concentration aerobically, unlike D-lactate, which formed in higher amounts anaerobically. Storage in air was the worst packaging method, resulting in greening and unpleasant off-odours associated with the high acetate content of the sausages. Carbon dioxide had no significant effect on extending shelf-life. The factors affecting the natural selection of Lact. sakei/curvatus in taverna sausage are discussed. Moreover, it was attempted to correlate the metabolic activity of this group with the physicochemical changes and the spoilage phenomena occurring in taverna sausage under the different storage conditions.     

Evaluating rRNA as an indicator of microbial activity in environmental communities: limitations and uses

Microbes exist in a range of metabolic states (for example, dormant, active and growing) and analysis of ribosomal RNA (rRNA) is frequently employed to identify the ‘active'' fraction of microbes in environmental samples. While rRNA analyses are no longer commonly used to quantify a population''s growth rate in mixed communities, due to rRNA concentration not scaling linearly with growth rate uniformly across taxa, rRNA analyses are still frequently used toward the more conservative goal of identifying populations that are currently active in a mixed community. Yet, evidence indicates that the general use of rRNA as a reliable indicator of metabolic state in microbial assemblages has serious limitations. This report highlights the complex and often contradictory relationships between rRNA, growth and activity. Potential mechanisms for confounding rRNA patterns are discussed, including differences in life histories, life strategies and non-growth activities. Ways in which rRNA data can be used for useful characterization of microbial assemblages are presented, along with questions to be addressed in future studies.     

Behaviour of Listeria monocytogenes and autochthonous flora on meat stored under aerobic, vacuum and modified atmosphere packaging conditions with or without the presence of oregano essential oil at 5 degrees C

The effect of aerobic, modified atmosphere packaging (MAP; 40% CO2/30% O2/30% N2) and vacuum packaging (VP) on the growth/survival of Listeria monocytogenes on sterile and naturally contaminated beef meat fillets was studied in relation to film permeability and oregano essential oil. The dominant micro-organism(s) and the effect of the endogenous flora on the growth/survival of L. monocytogenes were dependent on the type of packaging film. The fact that L. monocytogenes increased whenever pseudomonads dominated, i.e. aerobic storage and MAP/VP in high-permeability film, and even earlier than on sterile tissue, suggests that this spoilage group enhanced growth of the pathogen. Brochothrix thermosphacta constituted the major proportion of the total microflora in MAP/VP within the low-permeability film, where no growth of L. monocytogenes was detected either on naturally contaminated or sterile meat fillets. The addition of 0.8% (v/w) oregano essential oil resulted in: (i) an initial reduction of 2-3 log10 of the majority of the bacterial population, with lactic acid bacteria and L. monocytogenes indicating the most apparent decrease in all gaseous environments, and (ii) limited growth aerobically and survival/death of L. monocytogenes in MAP/VP, regardless of film permeability.     

To survive or to slay: Resource-foraging role of metabolites implicated in allelopathy

The ecological relevance of allelopathy is highly debated due to the lack of phytotoxic concentrations of allelochemical in natural field conditions. Most of the putative allelochemicals are exuded at low concentrations, and subsequently undergo rapid chemical and biological degradation in soil matrices. At sub-toxic concentrations, due to hormesis effect, these compounds could possibly have a stimulatory effect on plant growth. Many of the suggested allelopathic compounds are chelants and can complex-with and mobilize metal ions in soil. These complexation reactions will detoxify the compound, but will increase the chemical-nutrient-foraging ability of the donor plant. The concentration in which these compounds are exuded matches with other similar secondary metabolites facilitating plant nutrient acquisition. Irrespective of whether the implicated PSMs facilitate donor plant in chemical nutrient-foraging or in poisoning the neighbors, the conferred advantage translates in terms of resource availability—in first case the donor enjoys uncontested nutrient uptake efficiency, where as in the latter the donor gain an uncontested access to resources. This further reaffirms the notion that resource competition and allelopathy are inextricable. Since most of the secondary metabolites could mobilize nutrients from soil, along with its phytotoxic effect, complementary self-facilitation roles of these compounds should be investigated.Key words: allelopathy, allelochemical, catechin, 8-hydoxyquinoline, hormesis, competition, resource foraging, chelant, plant invasion, phytosiderophorePlants are shown to produce a diverse array of secondary metabolites under various environmental conditions.¹ Some of these compounds elicit specific responses in other organisms, and may increase plant fitness, especially by defending plants against herbivores² or through toxicity to competitors.³ Allelopathy is plant secondary metabolite (PSM) mediated (predominantly negative) plant-to-plant interaction. With the advancement of analytical chemistry, coupled with our understanding in cellular and molecular biology, recently, a vast array of secondary compounds have been proposed to have allelochemical properties. This includes compounds identified from cultivated crops⁴ to invasive exotic plants.⁵ Allelopathy has been employed successfully in agricultural systems, where cover crops, following tissue maceration and incorporation into soil that releases PSM, have been successfully employed for resident pest control.⁴ In contrast, the significance of allelopathic interactions in natural ecosystems is highly debated; the main drawback being lack of sufficient concentrations of the PSM to elicit a phytotoxic response under field conditions.^6–11 The present article addresses the dilemma of allelopathic research in natural systems, and excludes agricultural systems.     

Growth limitation in hybridoma cell cultures: The role of inhibitory or toxic metabolites

Hybridoma cells usually grow to fairly low cell densities in batch cultures (1–3×10⁶ cells/ml). The reason for this is either that essential nutritional components of the medium are consumed, or that the cells produce some kind of inhibitory or toxic metabolite. This investigation presents evidence for the latter. Spent medium from cultures of hybridoma cells did not support growth of cells at lower cell densities (1–3×10⁵ cells/ml). The ability to support cell growth could not be restored by adding additional serum, energy sources (glucose, pyruvate) or L-glutamine. Furthermore, the consumption of amino acids could not account for this growth inhibition. On the contrary, the spent medium contained a substance that inhibited cell growth. This substance or metabolite was found in a fraction eluted from a gel filtration column when spent medium was applied to the column. This substance was found in the spent medium from all hybridoma and myeloma cell lines that were tested. The molecular weight of the substance was about 5 kD. Spent medium from two hybridoma cell lines also contained a substance that was eluted in the same fraction as albumin (67 kD). It is likely that this (or these) substance(s) is responsible for the growth limitation in hybridoma cell cultures.Abbreviations PBS phosphate buffered saline     

Rhizobia and bradyrhizobia under salt stress: possible role of trehalose in osmoregulation

Seven rhizobium fredii strains and seven Bradyrhizobium japonicum strains were grown in defined medium with or without 20m m trehalose in the presence or absence of NaCl. Trehalose had no effect on the growth rate of the strains in the absence of NaCl, but increased the growth rate of some strains in the presence of NaCl. Bradyrhizobium japonicum strain RCR 3827 was completely inhibited by 0·08 m NaCl in absence of trehalose, but multiplied when trehalose was added. The results indicate that trehalose may act as an osmoregulator in these strains of Rhizobium and Bradyrhizobium .     

The cytochrome composition of the meat spoilage bacterium Brochothrix thermosphacta: identification of cytochrome a 3- and d-type terminal oxidases under various conditions

Brochothrix thermosphacta, grown in batch culture in a yeast-dextrose broth, at temperatures from 30 °C to 10 °C, contained diverse membrane-bound respiratory cytochromes. Under conditions of moderate aeration, cytochromes of the a-, b- and d-type were detected at all growth temperatures, but the proportions changed as a function of temperature, with the spectra of cells grown at 10 or 15 °C being dominated by a-type cytochrome(s). Cytochrome a ₃ was detected by its reactions with CO and cyanide in cells from all growth conditions. An additional cytochrome a, which was not cyanide-reactive, was also detected, suggesting the presence of an aa ₃ oxidase complex. Cytochrome d was cyanide- and CO-reactive, but not detectable in photodissociation spectra, presumably because of the very rapid recombination of CO at the sub-zero temperatures used. Decreasing the oxygen transfer rates to batch cultures resulted in enhanced expression of cytochrome d and changed the proportion of the aa ₃-type oxidase that could be attributed to ligand-binding cytochrome a ₃; at the lowest oxygen transfer rates, no cytochrome a was detected, suggesting the presence of a cytochrome ba ₃ terminal oxidase complex. Intact cells showed no evidence of a c-type cytochrome and no haem C was detected in membrane preparations. After growth at 10°C, the cytochrome composition of B. campestris was essentially identical to that of B. thermosphacta. The multiplicity of putative terminal oxidases in B. thermosphacta is discussed.     

Acyclic tetraprenyltoluquinols from Cystoseira sauvageuana and their possible role as biogenetic precursors of the cyclic cystoseira metabolites

From the brown alga Cystoseira sauvageuana two novel tetraprenyltoluquinols with an acyclic terpenoid side chain have been isolated. Their chemical transformation into compounds containing a cyclopentane ring provides a clue to the biogenesis of the cyclic Cystoseira metabolites.