Nutrition of Coryneform Bacteria from Milk and Dairy Sources

S ummary . Growth requirements were determined for 112 cultures of coryneform bacteria including strains of Corynebacterium bovis, C. ulcerans and C. lacticum ; in general they were quite distinctive. Of 42 C. bovis strains all had an unsaturated fatty acid requirement fulfilled in chemically defined media by Tween 80, and, but for an atypically nonureolytic strain, all utilized ammonia and urea nitrogen. Whereas vitamins were unessential for the majority of C. bovis. 18 strains failed to grow in the absence of nicotinic acid; however, for 11 of these Casamino Acids replaced nicotinic acid in supporting growth. Thirteen C. ulcerans strains from aseptically drawn milk, as well as two Type Culture strains (NCTC 7907 and 7908) from the human throat required amino acid nitrogen sources, nicotinic acid, pantothenate and biotin. Some nutritional heterogeneity was shown by 46 strains collectively described as C. lacticum obtained from market milk, milk products and dairy utensils; nevertheless the nutritional differences between the groups of these strains correlated broadly with other distinguishing properties. A group of 9 markedly caseolytic strains required amino acid nitrogen sources, thiamine and biotin, whereas a second group of 30 strains, some utilizing inorganic nitrogen sources, required thiamine and pantothenate, with or without biotin. The remaining C. lacticum strains together with a number of unidentified strains from similar sources were generally slow growing and had widely differing growth requirements.     

Chlortetracycline and Oxytetracycline Residues in Poultry Tissues and Eggs

A pad-plate method of assaying residual amounts of chlortetracycline (CTC) and oxytetracycline (OTC) using Bacillus cereus 213 was used to determine amounts of antibiotic left in tissues and eggs of poultry fed 1,000 and 200 ppm of CTC and OTC in basal feed mixtures. The effects of various methods of cooking the tissues and eggs and the potentiating effect of terephthalic acid (TPA) were studied. It was found that normal methods of roasting, frying, and autoclaving poultry tissue destroyed all residual CTC and OTC, even with the potentiating effect of TPA. The largest amounts of residual antibiotic were found in the liver, then breast, and then thigh tissue when assayed for CTC. Tissue assays for CTC revealed that it was not taken up as extensively as CTC and the largest amounts were found in the liver, then breast. OTC residue was seldom found in the thigh tissue. Terephthalic acid in 0.5% concentration increased the concentration found in all cases. Cooking by poaching and scrambling eggs did not destroy the antibiotic in all cases.     

Diversity among Opine-Utilizing Bacteria: Identification of Coryneform Isolates

Bacteria were isolated from soil and crown gall tumors by selection in minimal medium with an opine, such as succinamopine or mannopine, as the sole carbon source. The isolates were characterized for the pattern of opine utilization and identified. They were classified as mannityl opine or imino diacid utilizers and exhibited specificity of utilization similar to that described previously for Agrobacterium species. A minority of isolates were gram negative and were identified as Agrobacterium or Pseudomonas species; most were gram positive and belonged to the coryneform group. These results indicate that any specific effect of opines on the ecology of Agrobacterium tumefaciens is modulated by activities of other types of soil- and plant-associated bacteria.     

Possible Role of Coryneform Bacteria in Age Gelation of Ultrahigh-Temperature-Processed Milk

Two strains of psychrotrophic, gram-positive, proteolytic bacteria isolated from raw milk were identified as Arthrobacter or closely related coryneform species. We inoculated raw-milk samples with the strains (one strain per sample) and compared rates of gelation after ultrahigh-temperature processing with that of ultrahigh-temperature-processed controls. The trial indicated that either organism could play a role in age gelation of ultrahigh-temperature-processed milk.     

Lipids in the Classification and Identification of Coryneform Bacteria Containing Peptidoglycans Based on 2, 4-diaminobutyric Acid

Strains of 2, 4-diaminobutyric acid-containing coryneform bacteria were degraded by acid methanolysis and the non-hydroxylated fatty acid esters released examined by thin-layer and gas chromatography. The major fatty acid structural types were straight-chain, anteiso - and iso -methyl branched-chain acids. Polar lipids of the test strains were examined by two-dimensional thin-layer chromatography. All strains possessed very characteristic polar lipid patterns consisting of diphosphatidylglycerol, phosphatidylglycerol and a number of uncharacterized glycolipids. Menaquinones (vitamin K) were the sole isoprenoid quinones detected in the test strains. Corynebacterium insidiosum, Cor. michiganense, Cor. nebraskense and Cor. sepedonicum contained unsaturated menaquinones with nine isoprene units, whereas unsaturated menaquinones with 10 isoprene units predominated in strains of Cor. iranicum and Cor. tritici and a strain labelled Arthrobacter sp. The single strain of Cor. aquaticum examined contained comparable amounts of menaquinones with 10 and 11 isoprene units whereas strains of Cor. mediolanum and Flavobacterium dehydrogenans contained major amounts of menaquinones with 11 and 12 isoprene units. The results of the present study indicate that lipid markers may be of considerable value in the classification and identification of 2, 4-diaminobutyric acid-containing phytopathogenic and saprophytic coryneform bacteria.     

Fate of Bacteria in Chicken Meat During Freeze-Dehydration, Rehydration, and Storage

Total plate counts were determined on boneless cooked, cubed chicken meat obtained from a commercial processor. Survival of the natural flora was determined after the meat was freeze-dehydrated and rehydrated at room temperature for 30 min and 50, 85, and 100 C for 10 min. Total counts of bacteria in the rehydrated samples were determined during storage of the meat at 4, 22, and 37 C until spoilage odor was detectable. Meat samples were inoculated with Staphylococcus aureus, then dried, rehydrated, and stored at the same temperatures. Numbers of surviving organisms in the inoculated samples were determined with use of both selective and nonselective media. Representative genera surviving the various rehydration treatments were determined. Approximately 32% of the bacteria in the meat survived during dehydration and rehydration at room temperature. Many numbers and types of vegetative bacteria also survived rehydration at 50 C. When meat was rehydrated at 85 or 100 C, the initial count was less than one per gram. The only organisms isolated from samples rehydrated at 85 or 100 C were of the genus Bacillus. S. aureus in inoculated samples survived dehydration and rehydration at 60 C. Storage of all rehydrated samples at 4 C gave a good shelf life (18 or more days). The study indicates that freeze-dehydrated meat should be produced with adequate microbiological control and that such meat should be rehydrated in very hot water.     

Extracellular Enzymic Activity of Poultry Spoilage Bacteria

The extracellular enzymic activity has been studied of 224 strains of bacteria isolated mainly at 1° from spoiling chickens and turkeys and from poultry processing plants. The isolates comprised 44 strains of pigmented Pseudomonas , 57 strains of nonpigmented Pseudomonas , 29 strains of Ps. putrefaciens , 50 strains of oxidase positive Acinetobacter and 44 strains of oxidase negative Acinetobacter. None of the strains showed any significant activity against dextrin, starch, glycogen, inulin, dextran, xylan or pectin. Proteolytic activity was found mainly amongst 2 groups of pigmented pseudo-monads, and Ps. putrefaciens. Nuclease activity was found particularly amongst strains of Ps. putrefaciens and the oxidase negative Acinetobacter strains isolated from spoiling poultry. Almost all of the strains showed lipolytic activity when tested with tributyrin and a proportion of strains could also attack chicken fat. This latter property was particularly evident amongst the nonpigmented Pseudomonas strains.