Clostridium frigidicarnis sp. nov., a psychrotolerant bacterium associated with 'blown pack' spoilage of vacuum-packed meats.
Two strains of a psychrotolerant Clostridium, isolated from vacuum-packed, temperature-abused beef, were characterized using a multiphasic approach. The strains were Gram-positive motile rods producing elliptical subterminal spores during early stationary growth phase. The strains were psychrotolerant. At pH 7.0, they grew between 3.8 and 40.5 degrees C; their optimum growth temperature was 30.0-38.5 degrees C. At 30 degrees C, the pH range for growth was between 4.7 and 9.5; the optimum pH for growth was 6.4-7.2. The organisms were proteolytic and saccharolytic, lecithinase-positive and hydrolysed gelatin. The fermentation products formed in peptone/yeast extract/glucose/starch broth were acetate, ethanol, butyrate, isovalerate, butanol, isobutyrate, oxalacetate, lactate, hydrogen and carbon dioxide. The DNA G + C compositions of the two meat strains were 27.3 and 28.4 mol%. Phylogenetic analyses indicated that the strains belong to Cluster I of the genus Clostridium (sensu Collins et al., 1994). The new strains differed from phylogenetically related clostridia in terms of cellular fatty acid composition, soluble protein profiles and phenotypic properties. On the basis of phenotypic and genotypic characterization data, the strains were assigned to a new species for which the name Clostridium frigidicarnis is proposed; strain SPL77AT (= DSM 12271T) is the type strain. (+info)
Effects of a return chewing gum/packaging material mixture on in situ disappearance and on feed intake, nutrient digestibility, and ruminal characteristics of growing steers.
In situ and in vivo digestibility experiments were conducted to determine the acceptability, digestibility, and safety of a return chewing gum/packaging (G/P) material mixture when fed to steers. In the in situ experiment, both ruminal and intestinal disappearances were measured. Two ruminally and duodenally cannulated steers, which were given free access to alfalfa hay (AH), were used in this study. Duplicate Dacron bags containing the G/P were incubated in the rumen for 0, 3, 6, 12, 24, and 48 h. After ruminal incubation, the 12-, 24-, and 48-h bags were placed in the duodenum and collected in the feces to determine intestinal disappearance. In situ ruminal DM disappearance was greater than 70% for all substrates tested at 0 h, indicating high solubility of the substrates in water, and began to reach a plateau after 12 h of incubation. Intestinal in situ disappearance was not different (P>.25) from zero. In the digestion trial, four ruminally cannulated steers (337+/-21.3 kg BW; mean +/- SD) were used in a 4x4 Latin square design with the following treatments: 0) 50% corn (C), 50% AH; 10) 45% C, 45% AH, 10% G/P; 20) 40% C, 40% AH, 20% G/P; 30) 35% C, 35% AH, 30% G/P. Steers fed G/P-containing diets had greater (P<.01) DMI than the control steers. Increasing the G/P resulted in a linear (P<.05) increase in DMI. Apparent DM digestibility tended to be higher (P<.10) for the G/P-containing diets than for the control. A quadratic effect (P<.05) on digestible DMI was observed, with greater (P<.01) digestible DMI values for G/P-containing diets (4.8 vs. 5.8 kg/d). Digestible organic matter and total nonstructural carbohydrate intakes followed trends similar to those of DM. Apparent aluminum digestibility of G/P-containing diets was not different (P>.13) from zero. The level of G/P in the diet had no effect (P>.2) on total VFA concentration or ruminal pH. There was a linear decrease (P<.01) in the molar percentage of isobutyrate and isovalerate in addition to a linear increase (P<.01) in butyrate and valerate with increasing levels of G/P. There was a quadratic effect (P<.01) on molar proportions of acetate and propionate and on the acetate:propionate ratio. Results of both experiments suggest that G/P may be fed to safely replace up to 30% of corn-alfalfa hay diets for growing steers with advantages in improving DMI and digestibility. (+info)
Clostridium algidixylanolyticum sp. nov., a psychrotolerant, xylan-degrading, spore-forming bacterium.
A psychrotolerant Clostridium species was isolated from vacuum-packed, temperature-abused raw lamb. Colonies of this micro-organism on sheep-blood agar were circular with an entire margin, grey-white, translucent and beta-haemolytic. Cells were single, tapered, motile rods. Elliptical subterminal spores were produced in the late stationary growth phase. Spores did not cause swelling of the maternal cells. The micro-organism was obligately anaerobic. In peptone yeast extract glucose starch (PYGS) broth at pH 7.0, the micro-organism grew optimally between 25.5 and 30.0 degrees C. The temperature range for growth was 2.5-32.2 degrees C. At 26 degrees C, the micro-organism grew optimally at pH 6.8 to 7.0. The pH range for anaerobic growth was 4.7-9.1. The micro-organism was saccharoclastic, hydrolysed starch and degraded xylan. The fermentation products formed in PYGS broth were acetate, formate, lactate, ethanol, butyrate, butanol, hydrogen and carbon dioxide. The G + C content of the DNA was 38.4 mol%. Phylogenetic analyses indicated that the strain belongs to cluster XIVa of the genus Clostridium (sensu Collins et al. 1994). The new strain differed from phylogenetically related clostridia in terms of cellular fatty acid composition, soluble protein profiles and phenotypic properties. On the basis of phenotypic and genotypic characterization data, the strain was assigned to a new species, namely Clostridium algidixylanolyticum. The type strain is strain SPL73T (= DSM 12273T). (+info)
Clostridium gasigenes sp. nov., a psychrophile causing spoilage of vacuum-packed meat.
Two psychrophilic Clostridium strains, DB1AT and R26, were isolated from incidences of 'blown-pack' spoilage of vacuum-packed chilled lamb. Vacuum packs of meat inoculated with these strains developed gas bubbles and pack distension within 14 d storage at 2 degrees C. The two main gases responsible for pack distension were carbon dioxide and hydrogen. 1-Butanol, butyric and acetic acid and butyl esters were the major volatile compounds produced by the strains in the artificially inoculated packs. The unknown strains were Gram-positive motile rods producing elliptical subterminal spores during the late-stationary growth phase. At pH 7.0, they grew from -1.5 to 26 degrees C, and their optimum growth temperature was 20-22 degrees C. At 20 degrees C, the pH range for growth was 5.4-8.9 and the optimum pH for growth was 6.2-8.6. In peptone/yeast extract broth, the organisms grew little or not at all in the absence of fermentable carbohydrates. Both strains hydrolysed gelatin, aesculin and starch. The fermentation products formed in peptone yeast extract glucose starch broth were ethanol, acetate, butyrate, lactate, butanol, carbon dioxide and hydrogen. The G+C contents of the DNA of strains DB1AT and R26 were 29.4 and 28.3 mol%, respectively. Phylogenetic analyses indicated that the strains belong to cluster I of the genus Clostridium (sensu Collins et al. 1994). The new strains differed from the phylogenetically related clostridia in cellular fatty acid composition, soluble protein profiles and phenotypic properties. On the basis of rDNA analysis and phenotypic and phylogenetic characterization, the strains were assigned to a new species for which the name Clostridium gasigenes is proposed. Strain DB1AT (= DSM 12272T) is designated as the type strain. (+info)
Lactobacillus algidus sp. nov., a psychrophilic lactic acid bacterium isolated from vacuum-packaged refrigerated beef.
Lactobacillus algidus sp. nov. is described on the basis of 40 strains isolated as one of the predominant bacteria from five specimens of vacuum-packaged beef collected from different meat shops and stored at 2 degrees C for 3 weeks. These strains were quite uniform in the overall characteristics examined. They are facultatively anaerobic, psychrophilic, Gram-positive, non-spore-forming, non-motile, lactic acid-homofermentative rods. The cells occurred singly and in pairs on agar media and in rather long chains in broth media. They differed in several cultural and biochemical characteristics from the authentic meso-diaminopimelic acid-positive or psychrophilic lactic acid bacteria in the genera Lactobacillus, Carnobacterium and Brochothrix. The SDS-PAGE whole-cell protein pattern was clearly distinctive. DNA-DNA hybridization and phylogenetic analysis of 16S rDNA also failed to associate these strains closely with any of the validly described organisms used. The phylogenetic analysis showed that these strains are rather remotely but most closely related to Lactobacillus mali (93% sequence similarity), which belongs to the Lactobacillus casei/Pediococcus group. Therefore, these strains should be included in the genus Lactobacillus and considered to represent a new species, Lactobacillus algidus sp. nov. The type strain is M6A9T (= JCM 10491T). (+info)
Applicability of an Arrhenius model for the combined effect of temperature and CO(2) packaging on the spoilage microflora of fish.
The temperature behavior of the natural microflora on the Mediterranean fish red mullet (Mullus barbatus) was examined as a case study. The growth of the spoilage bacteria Pseudomonas spp., Shewanella putrefaciens, Brochothrix thermosphacta, and lactic acid bacteria was modeled as a function of temperature and the concentration of carbon dioxide in modified atmosphere packaging. Combined models were developed and comparatively assessed based on polynomial, Belehradek, and Arrhenius equations. The activation energy parameter of the Arrhenius model, E(A), was independent of the packaging atmosphere and ranged from 75 to 85 kJ/mol for the different bacteria, whereas the preexponential constant decreased exponentially with the packaging CO(2) concentration. We evaluated the applicability of the models developed by using experimental bacterial growth rates obtained from 42 independent experiments performed with three Mediterranean fish species and growth rates predicted from the models under the same temperature and packaging conditions. The accuracy factor and bias factor were used as statistical tools for evaluation, and the developed Arrhenius model and the Belehradek model were judged satisfactory overall. (+info)
Characterization of Leuconostoc gasicomitatum sp. nov., associated with spoiled raw tomato-marinated broiler meat strips packaged under modified-atmosphere conditions.
Lactic acid bacteria (LAB) associated with gaseous spoilage of modified-atmosphere-packaged, raw, tomato-marinated broiler meat strips were identified on the basis of a restriction fragment length polymorphism (RFLP) (ribotyping) database containing DNAs coding for 16S and 23S rRNAs (rDNAs). A mixed LAB population dominated by a Leuconostoc species resembling Leuconostoc gelidum caused the spoilage of the product. Lactobacillus sakei, Lactobacillus curvatus, and a gram-positive rod phenotypically similar to heterofermentative Lactobacillus species were the other main organisms detected. An increase in pH together with the extreme bulging of packages suggested a rare LAB spoilage type called "protein swell." This spoilage is characterized by excessive production of gas due to amino acid decarboxylation, and the rise in pH is attributed to the subsequent deamination of amino acids. Protein swell has not previously been associated with any kind of meat product. A polyphasic approach, including classical phenotyping, whole-cell protein electrophoresis, 16 and 23S rDNA RFLP, 16S rDNA sequence analysis, and DNA-DNA reassociation analysis, was used for the identification of the dominant Leuconostoc species. In addition to the RFLP analysis, phenotyping, whole-cell protein analysis, and 16S rDNA sequence homology indicated that L. gelidum was most similar to the spoilage-associated species. The two spoilage strains studied possessed 98.8 and 99.0% 16S rDNA sequence homology with the L. gelidum type strain. DNA-DNA reassociation, however, clearly distinguished the two species. The same strains showed only 22 and 34% hybridization with the L. gelidum type strain. These results warrant a separate species status, and we propose the name Leuconostoc gasicomitatum sp. nov. for this spoilage-associated Leuconostoc species. (+info)
Rapid, quantitative PCR monitoring of growth of Clostridium botulinum type E in modified-atmosphere-packaged fish.
A rapid, quantitative PCR assay (TaqMan assay) which quantifies Clostridium botulinum type E by amplifying a 280-bp sequence from the botulinum neurotoxin type E (BoNT/E) gene is described. With this method, which uses the hydrolysis of an internal fluoregenic probe and monitors in real time the increase in the intensity of fluorescence during PCR by using the ABI Prism 7700 sequence detection system, it was possible to perform accurate and reproducible quantification of the C. botulinum type E toxin gene. The sensitivity and specificity of the assay were verified by using 6 strains of C. botulinum type E and 18 genera of 42 non-C. botulinum type E strains, including strains of C. botulinum types A, B, C, D, F, and G. In both pure cultures and modified-atmosphere-packaged fish samples (jack mackerel), the increase in amounts of C. botulinum DNA could be monitored (the quantifiable range was 10(2) to 10(8) CFU/ml or g) much earlier than toxin could be detected by mouse assay. The method was applied to a variety of seafood samples with a DNA extraction protocol using guanidine isothiocyanate. Overall, an efficient recovery of C. botulinum cells was obtained from all of the samples tested. These results suggested that quantification of BoNT/E DNA by the rapid, quantitative PCR method was a good method for the sensitive assessment of botulinal risk in the seafood samples tested. (+info)