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The effect of evaporative air chilling on quality of fresh chicken carcasses was compared with air chilling as reference method. Cooling efficiency and total heat loss were significantly higher for evaporative air chilling. The chilling method was of great importance for weight loss. Chicken chilled in cold air lost considerably more weight than chicken cooled by evaporative air chilling; the difference was 1.8%. The chilling method also affected the skin color and the amount of moisture on skin surface. After evaporative air chilling, the chicken carcasses had a lighter color and more water on the back and under the wings. The moisture content in skin and meat, cooking loss, and pH were not affected by chilling method. Odor attributes of raw chicken and odor and flavor attributes of cooked chicken did not show any significant differences between the two chilling methods. The shelf life of chicken stored at 4 and -1 C were not affected significantly by chilling method. Storage time and temperature appeared to be the decisive factors for sensory and microbiological quality of fresh chicken carcasses.

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Results of carbon source assimilation tests (17 carbon compounds) led to 88% of pseudomonads from cold-stored chicken carcasses being assigned to one of 17 groups. Of these groups, 13 had combinations of properties identical to, or with readily recognizable degrees of similarity to those of published species/biovars. Two of the four groups having carbon assimilation patterns dissimilar to any known species had cellular fatty acid composition corresponding to Pseudomonas fluorescens, and two to Pseudomonas lundensis or Pseudomonas fragi. The P. fluorescens biovars all had higher amounts of 16:1 cis 9 (21-37%) and 18:1 cis 11 (10-19%), than of 17:0 cyclo (1-17%) and 19:0 cyclo (0-1%). In contrast, for P. lundensis and P. fragi, the relative amounts of these unsaturated acids and cyclopropane acids were reversed. Both the carbon source assimilation tests and the cellular fatty acid composition led to the conclusion that none of the species were dominant, although the P. fluorescens biovars constituted about 50% of the isolated pseudomonads.

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Although sensory and/or microbiological analyses are widely relied on when assigning shelf-life of foods or trouble shooting problems with spoilage under storage, they do have drawbacks. Delay in obtaining results is one of them. The expense of the expert panels required to obtain meaningful sensory evaluations is another, while spoilage is not always of microbial origin. Even when it is, there are an increasing number of situations, including that of meats and fish packaged in modified atmospheres, where the relationships between microbial growth and spoilage onset is poorly defined. Chemical analysis has long been recognized as a means of circumventing at least some of the drawbacks and its potential is reviewed below. From the data presented it can be concluded that chemical characterization of spoilage processes is presently of most value in trouble shooting i.e. establishing the causes of spoilage. Its value in assigning total or remaining shelf-life requires more knowledge of the chemical processes leading to reduced acceptability/spoilage and of their correlations with sensory and microbiological changes.

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A Clostridium species causing spoilage of vacuum-packed refrigerated pork was isolated and characterized. The unknown organism differed phenotypically from other clostridial species usually associated with spoilage. Phylogenetic analyses based on 16S rRNA gene sequencing demonstrated that the psychrotroph represents a distinct line of descent within the genus Clostridium. It is proposed that the organism be classified as a new species of the genus Clostridium, Clostridium algidicarnis.

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Oligonucleotide probes specific for Carnobacterium and Leuconostoc species were constructed from the variable regions of 16S rRNA obtained from the literature and sequence data bases. The probes were hybridized with crude nucleic acid extract from 32 type strains of lactic acid bacteria (LAB) commonly found on meat. Two of the probes hybridized only to the four Carnobacterium species whereas the other two hybridized only to five of the six Leuconostoc species tested. The probes were also hybridized with nucleic acids from unknown strains of LAB. The identification was consistent with the results of biochemical tests used to characterize the two genera.

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Taxonomic studies were performed on an anaerobic Gram-positive, spore-forming, psychrophilic bacterium originally isolated from spoiled vacuum-packed refrigerated beef. Based on the present finding it is proposed that this unknown psychrophilic bacterium be classified as a new species of the genus Clostridium, as Clostridium estertheticum sp. nov. The type strain is NCIMB 12511.

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Volatile compounds produced by Pseudomonas fragi and mixed, natural floras on beef of normal pH (5.5-5.8; glucose greater than 1500 micrograms/g) and high pH (6.3-6.8; glucose less than 10 micrograms/g) included a range of alkyl esters and a number of sulphur-containing compounds including dimethylsulphide but not hydrogen sulphide. Production of the last was a property common to the other Gram-negative organisms tested viz. Hafnia alvei, Enterobacter agglomerans, Serratia liquefaciens, Alteromonas putrefaciens and Aeromonas hydrophila, all of which produced similar off-odours and, with the exception of E. agglomerans, 'greening' on high pH meat. Serratia liquefaciens also produced greening of normal pH meat. Acetoin and diacetyl were major end products of Brochothrix thermosphacta but the related 2,3-butanediol was formed only on normal pH meat. The Enterobacteriaceae produced the same compounds but only on normal pH meat and together with Br. thermosphacta were probable sources of these compounds and of the free and esterified branched-chain alcohols detected in the naturally contaminated samples.

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The amine content of fresh and vacuum-packaged beef of normal pH stored at 1 degree C was evaluated by high performance liquid chromatography of dansyl derivatives. Fresh samples contained five amines, viz. putrescine, cadaverine, histamine, spermine and spermidine. Development of a natural spoilage flora during storage led to increases in concentration of putrescine and cadaverine and the production of a sixth amine, tyramine. Pure culture meat inoculation experiments showed tyramine formation to be restricted to lactobacilli and to strains of Lactobacillus divergens and Lact. carnis in particular; strains of leuconostocs, Enterobacteriaceae, Pseudomonas spp. and Brochothrix thermosphacta were negative. Production of tyramine at cell densities less than log10 6/cm2 indicated its potential as an objective measure of acceptability/spoilage.

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Volatile compounds produced by 31 strains of pseudomonads and by reference strains of Pseudomonas fragi and Ps. fluorescens biotype 1 during growth on beef stored at 6 degrees C in air were analysed by gas chromatography-mass spectrometry of headspace gases. Compounds of major sensory significance were ethyl and methyl esters of C2-C8 fatty acids and sulphur-containing compounds which included methane- and isopropanethiols and their related sulphides and thioesters but not hydrogen sulphide. Ester production was mainly associated with growth of some, but not all, Ps. fragi and related meat strains but sulphur-containing compounds were produced by all but a single meat strain. A minority of other meat strains produced greater amounts of methyl ketones, secondary alcohols and unsaturated hydrocarbons believed to be of lipid origin.

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Of the meat strains of streptobacteria, leuconostocs, Enterobacteriaceae and Brochothrix thermosphacta tested, only Hafnia alvei and Serratia liquefaciens showed diamine-producing potential during growth in pure culture on beef stored in vacuum packs at 1 degree C. Both organisms produced cadaverine at concentrations similar to those reported previously in naturally contaminated beef stored under the same conditions. Putrescine concentrations produced by the two organisms, however, were an order of magnitude lower. During the growth on beef of either H. alvei or S. liquefaciens in mixed culture with arginine-utilizing strains of streptobacteria, putrescine as well as cadaverine concentrations were similar to those detected in naturally contaminated samples.

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The microbial flora of naturally contaminated beef stored in air was similar to that frequently recorded for meat stored under gas permeable films. Compounds produced as a result of microbial growth were acetoin, diacetyl, 3-methyl-1-butanol, 2-methyl-1-propanol, ethyl esters of acetic, propionic, butyric, isovaleric and hexanoic acids, methane thiol, dimethylsulphide, dimethyl disulphide, 1-undecene and 1,4-undecadiene. The first four compounds, which are known end-products of Brochothrix thermosphacta metabolism, were consistently detected at earlier stages of storage than the others, all of which have been shown to be produced by Pseudomonas spp. A pattern of odour development consistent with the chemical changes was also observed.

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Bacterial numbers, putrescine and cadaverine concentrations and pH were measured at regular intervals during the chill storage of vacuum packed beef. Odours on opening the packs were also assessed. Cadaverine concentration increased more rapidly than that of putrescine and measurable increases were evident before maximum bacterial numbers were attained and before any permanent off-odours were detected. Diamine concentrations correlated better with total viable count (TVC) than with counts of Gram negative organisms.

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Five strains representing four clusters of meat spoilage pseudomonads were grown on sterile beef at 5 degrees C. After 7 days incubation sensory assessments were made and the chemical composition of the headspace gases determined by gas chromatography-mass spectrometry. There was good correlation between odour descriptions and chemical data for three of the strains. The most numerous types of product were esters and sulphur-containing compounds. Of 45 compounds identified only 1-undecene was common to all the tested strains.

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Growth of the meat spoilage organism, Brochothrix thermosphacta, was supported by either glucose or glycerol with acetoin production. Similar yields of bacteria were obtained at growth-limiting concentrations of either substrate. The rate of uptake of each substrate was similar but glycerol supported higher rates of respiration than glucose. These findings are discussed in relation to metabolism of glycerol and the use of glycerol for meat storage.

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Four extracellular proteolytic enzymes (I-IV) (EC 3.4.22.-) were identified in static cultures of Chromobacterium lividum (NCIB 10926) by agar gel electrophoresis and isoelectric focusing. Proteinases I-III were freed of non-enzymic protein by chromatography on TEAE-cellulose and CM-cellulose. The enzyme mixture was then fractionated in a pH gradient by isoelectric focusing. All three enzymes were shown to be heat-labile metallo-enzymes. Optimal activity occurred at pH 5.6 for enzyme I and at pH 6.2 for enzymes II and III. Remazolbrilliant Blue-hide powder was a sensitive substrate for these enzymes. Proteinase I was also shown to degrade haemoglobin and casein effectively, but not myoglobin, ovalbumin or bovine serum albumin. Proteinases I-III exhibited molecular weight values of 75 000, 72 000 and 67 000 by exclusion chromatography and 71 000 and 66 000 by sodium dodecyl sulphate-poly-acrylamide-gel electrophoresis for enzyme I and II, respectively. The amino acid compositions of enzymes I and II were somewhat similar. Proteinase I was inhibited by EDTA, 1,2-di(2-aminoethoxy)ethane-N,N,N',N'-tetraacetic activity. Mg2+ could substitute for Ca2+ or Mn2+ for Co2+. The interrelationship of proteinases I-III is discussed.

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