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Changes in the microflora of irradiated petrale sole (Eopsetta jordani) fillets stored aerobically at 0.5 C.

The microfloral changes on irradiated petrale sole fillets during aerobic (packaged with oxygen-permeable film), refrigerated storage were determined by the identification of bacterial and yeast isolates to the generic level. The samples were irradiated at 0.0, 0.1, 0.15, 0.2, 0.3, and 0.4 Mrad by use of a cobalt-60 gamma source, were stored at 0.5 C, and were examined periodically for spoilage, total microbial population, and composition. The preirradiation flora of the fresh fillets consisted of coryneforms, Achromobacter, Micrococcus, Flavobacterium, Pseudomonas, and Lactobacillus. Immediately after irradiation, Micrococcus, Achromobacter, coryneforms, and Bacillus were predominant. The flora of the nonirradiated fillets at the time of spoilage consisted of Pseudomonas and Achromobacter. The flora of the irradiated fillets at the time of spoilage consisted of Achromobacter and Trichosporon.

Alcaligenes↗

Selection of microbial population in chlortetracycline-treated ocean perch (Sebastodes alutus).

Microorganisms isolated from chlortetracycline (CTC)-treated ocean perch were individually tested to determine their relative sensitivity to CTC by use of CTC-impregnated paper discs. The CTC sensitivity varied among isolates obtained from untreated fish. The proportion of CTC-resistant species increased with higher CTC concentration and the length of storage at 7 C. Among individual generic groups, "coryneforms" and yeasts were more resistant to CTC than other groups. Pseudomonas, Achromobacter, Flavobacterium, Bacillus, and Lactobacillus all contained species either resistant or sensitive to CTC. More CTC-resistant species in these genera accumulated with the increased CTC concentrations and with the length of storage in the presence of CTC.

Alcaligenes↗

Roles of microorganisms in the deterioration of rockfish.

Seven species of microorganisms indigenous to fish were inoculated into sterile fish muscle, obtained from Sebastodes alutus and Sebastalobus alascanus, and the production of total volatile acids (TVA) and total volatile bases (TVB) was determined during storage at 1 C. No production of TVA and TVB was noted in sterile fish, fish inoculated with Pseudomonas type III species (PIII-985), or fish inoculated with Flavobacterium (F-1070). The inactivity of PIII-985 was due to its low maximal growth level, whereas the inactivity of F-1070 was due to its slow growth rate. Pseudomonas type I species (PI-950), Pseudomonas type II species (PII-952), and two Achromobacter (Acinetobacter) species (A-981 and A-70) grew and actively produced TVA and TVB. TVB levels increased steadily even after the microbial number reached a maximum, whereas TVA levels declined toward the end of the storage period. It appeared that a minimal microbial growth level of 10(8) cells/g was needed before detectable levels of TVA or TVB could be produced. Regardless of genera, microorganisms which grew well in fish were able to produce TVA and TVB.

Alcaligenes↗

Microbiological evaluation of Pacific shrimp processing.

Microbiological evaluation of Pacific shrimp (Pandalus jordani) processing was made from samples obtained at five key processing points. The microbial count of raw shrimp ranged from 1.3 x 10(6) to 3.0 x 10(6). The initial microbial flora, in order of predominance, was Acinetobacter-Moraxella, Flavobacterium, Pseudomonas, gram-positive cocci, and Bacillus species. No yeasts were isolated. Differences in processing practices influenced both microbial count and the shrimp flora. The microbial load, however, always increased after peeling and sorting operations and decreased after cooking, washing, and brining steps. Significantly, the gram-positive cocci were recovered with increasing frequency after each processing step, reaching 76% of the total load in a final product. Most of them, however, were coagulase-negative.

Alcaligenes↗

Microbiological study of water-softener resins.

Microbial identification using effluents backflushed from exhausted urban and rural tank resins and cleaned resins containing the sulfonated copolymer of styrene and divinylbenzene (SDB) were completed, along with microbial assessment of the concentrated stock salt brine. Forty-four different bacterial and fungal genera were identified. Extensive biochemical and animal virulence tests completed on one of the six bacterial salt brine isolates indicated a pathogenic staphylococcal strain. The retention of Staphylococcus aureus, a Flavobacterium sp, and Escherichia coli B bacteriophage was demonstrated both by using the nonexhausted sodium-regenerated resin and by using the same resin exchanged with different mono-, di-, and trivalent cations. Effluent counts completed after bacterial seepage through the resins indicated the Pb(++) exchanged resin removed 55% of the bacteria; Na(+), Fe(++), and Al(+++) removed 31 to 36% and Ca(++) and Cu(++) removed about 10 to 15%. Seventy per cent or more of the bacteriophage was removed by Fe(++), Cu(++), and Al(+++), whereas the Ca(++) and Na(++) cations removed 25 to 31%. Over a 77-day period, nonsterile tap water was passed through bacterial seeded and uninoculated SDB (Na) resin columns. Effluent and resin elution counts demonstrated the growth and survival of 2 different bacteria per column. Increased bacterial retention, survival, and multiplication occurred concomitantly with accumulation of organic and inorganic materials and the Ca(++) and Mg(++) cations from the tap water. Furthermore, microbial elution from resin particles taken from column depths of 1, 8, and 16 cm indicated a bacterial diminution with increasing depths.

Aluminum↗

Control of growth rate by initial substrate concentration at values below maximum rate.

The hyperbolic relationship between specific growth rate, mu, and substrate concentration, proposed by Monod and used since as the basis for the theory of steady-state growth in continuous-flow systems, was tested experimentally in batch cultures. Use of a Flavobacterium sp. exhibiting a high saturation constant for growth in glucose minimal medium allowed direct measurement of growth rate and substrate concentration throughout the growth cycle in medium containing a rate-limiting initial concentration of glucose. Specific growth rates were also measured for a wide range of initial glucose concentrations. A plot of specific growth rate versus initial substrate concentration was found to fit the hyperbolic equation. However, the instantaneous relationship between specific growth rate and substrate concentration during growth, which is stated by the equation, was not observed. Well defined exponential growth phases were developed at initial substrate concentrations below that required for support of the maximum exponential growth rate and a constant doubling time was maintained until 50% of the substrate had been used. It is suggested that the external substrate concentration initially present "sets" the specific growth rate by establishing a steady-state internal concentration of substrate, possibly through control of the number of permeation sites.

Culture Media↗

Bacterial flora of the hemolymph of the blue crab, Callinectes sapidus: numerical taxonomy.

Bacteria isolated from the hemolymph of normal blue crabs were found to be predominantly Vibrio sp., Pseudomonas sp., Acinetobacter sp., Bacillus sp., Flavobacterium sp., and coliforms. Vibrio parahaemolyticus, a cosmopolitan facultative pathogen widely implicated in outbreaks of gastroenteritis related to the consumption of improperly processed seafood, was present in crabs collected between the months of May to November and was identified in up to 21% of the hemolymphs sampled.

Acinetobacter↗

Microflora and invert sugars in juice from healthy tissue of stored sugarbeets.

Bacterial populations increased in juice of healthy tissue of sugarbeet roots stored at 5 C. Average counts showed a sixfold increase after 150 days of storage. Invert sugar levels increased over threefold in "American 4 Hybrid A" and remained fairly constant in "Mono-Hy D-2." The former cultivar also had significantly higher bacterial colony counts than the latter before 90 days of storage. Of 36 isolates identified, 16 were Pseudomonas spp. including P. chlororaphis; 6 Bacillus spp. including B. subtilis; 5 Arthrobacter spp. including A. globiformis; 4 yeasts; 2 Erwinia spp; 2 Flavobacterium spp. including F. aquatile; and Streptomyces longisporus. Isolates of all genera except S. longisporus were able to hydrolyze sucrose in vitro.

Arthrobacter↗

Protection of mice from lethal endotoxemia by use of an ornithine-containing lipid or a serine-containing lipid.

The effects of an ornithine-containing lipid [alpha-N-(3-acyloxyacyl)-ornithine (Orn-L)] or a serine-containing lipid [alpha-N-(3-acyloxyacyl)-serine (Ser-L)] from Flavobacterium meningosepticum on lethal endotoxemia in mice were examined. When 500 micrograms of Orn-L was intravenously administered 1 h before intravenous administration of a lethal dose of endotoxin, none of the mice died. The protective effect of Ser-L was weaker than that of Orn-L. Light and electron microscopic studies demonstrated that necrosis of hepatocytes caused by endotoxin was prevented by pretreatment with Orn-L. Furthermore, Kupffer cells were activated morphologically 1 h after the administration of Orn-L or Ser-L, and the liposomes of the lipoamino acids were incorporated into phagolysosomes in activated Kupffer cells. The activity of tumor necrosis factor in sera of endotoxin-treated mice was decreased markedly by pretreatment of mice with Orn-L. In vitro, the lipoamino acids suppressed endotoxin-induced tumor necrosis factor generation but did not suppress tumor necrosis factor generation induced by zymosan and whole cells of Staphylococcus aureus. These results suggested that Orn-L and Ser-L can be used as specific blocking agents against endotoxin. The blocking mechanism may be antagonistic, because of the structural similarities between the lipoamino acids and endotoxin lipid A.

Animals↗

Nicotinamide deamidation by microorganisms in rat stomach.

We have extended our investigation of nicotinamide deamidation in the stomach of conventional rats. The bacterial species in the pars preventricularis were identified as Flavobacterium peregrinum, Escherichia coli, Streptococcus faecalis, and Lactobacillus acidophilus, listed in order of decreasing deamidase activity. Nicotinamide-7-(14)C ingested into rat stomach was rapidly deamidated to nicotinic acid. These results contribute to the accumulated evidence that microorganisms present in the pars preventricularis of rat stomach are responsible for the deamidation of nicotinamide to nicotinic acid, a known precursor of mammalian pyridine nucleotides.

Aminohydrolases↗

Surface-bound nuclease of Staphylococcus aureus: localization of the enzyme.

The cellular localization of staphylococcus nuclease, previously known as an exoenzyme, was investigated, and the following results were obtained. (i) When Staphylococcus aureus cells were converted to protoplasts by cell wall lytic enzyme L-11 (a bacteriolytic enzyme purified from Flavobacterium sp. which specifically hydrolyzes amide and peptide linkages of murein layers), over 80% of the cell-bound nuclease was released into the surrounding sucrose medium. (ii) The cell-bound nuclease was associated with the cell-wall membrane fraction of mechanically disrupted cells. (iii) The nuclease activity of cell-wall membrane fractions from cells during early and late stages of protoplast formation were compared. Less activity was found in the late stage. These results suggest that nuclease may be located at or near the surface of the cells. The distribution of cell-bound nuclease in the cell-wall membrane fraction varied with the growth conditions of S. aureus. The activity of alkaline phosphatase, another surface enzyme, was also investigated. Less of this enzyme than nuclease was released when the cells were converted to protoplasts.

Alkaline Phosphatase↗

Carboxypeptidase displaying differential velocity in hydrolysis of methotrexate, 5-methyltetrahydrofolic acid, and leucovorin.

An enzyme that catalyzes the hydrolysis of folic acid and the antifolate methotrexate nearly 20 times more rapidly than the hydrolysis of 5-methyltetrahydrofolate was extraced from a gram-negative bacterium tentatively identified as a Flavobacterium sp. The enzyme was purified 500-fold and found to have a molecular weight of about 53,000. Apparently a metallo-enzyme, it is inhibited by citrate and ethylenediaminetetraacetic acid (EDTA). Ca2+, Co2+, Mg2+, and Zn2+ reverse inhibition by EDTA, whereas Ca2+ and Zn2+ are weak activators in the absence of EDTA. The enzymatic reaction releases the carboxy-terminal glutamyl moiety of derivatives of pteroyl-mono-L-glutamic acid. Substituents on N5 of the pteridine ring decrease the velocity of hydrolysis. Some non-specificity for the terminal amino acid is expressed. The strikingly different rates of hydrolysis of methotrexate and 5-methyltetrahydrofolate have stimulated interest in this enzyme for its potential clinical value in improving the therapeutic index of methotrexate.

Carboxypeptidases↗

Bacteriophages of methanotrophic bacteria.

Bacteriophages of methanotrophic bacteria have been found in 16 out of 88 studied samples (underground waters, pond water, soil, gas and oil installation waters, fermentor cultural fluids, bacterial paste, and rumen of cattle) taken in different geographic zones of the Soviet Union. Altogether, 23 phage strains were isolated: 10 strains that specifically lysed only Methylosinus sporium strains, 2 strains that each lysed 1 of 5 Methylosinus trichosporium strains studied, and 11 strains that lysed Flavobacterium gasotypicum and, at the same time, 1 M. sporium strain. By fine structure, the phages were divided into two types (with very short or long noncontractile tails); by host range and serological properties, they fell into three types. One-step growth characteristics of the phages differed only slightly; the latent period varied from 6 to 8 h, the rise period varied from 4 to 6 h, and the average burst size was 100. All phages had guanine- and cytosine-rich double-stranded deoxyribonucleic acid consisting of common nitrogen bases. The molecular mass of the deoxyribonucleic acid as determined by restriction endonuclease analysis was 29.4 X 10(6) for M. sporium phages and 44 X 10(6) for F. gasotypicum phages. By all of the above-mentioned properties, all phages within each of the groups were completely identical to one another, but differed from phages of other groups. Bacteriophages lysing M. sporium and M. trichosporium GB2 were identical to phages M1 and M4, respectively, which were isolated earlier in the German Democratic Republic on the same methanotrophic species.

Antigens, Viral↗

Plasmid-determined enzymatic degradation of nylon oligomers.

The nylon oligomer (6-aminohexanoic acid cyclic dimer) degradation genes on plasmid pOAD2 of Flavobacterium sp. KI72 were cloned into Escherichia coli vector pBR322. The locus of one of the genes, the structural gene of 6-aminohexanoic acid linear oligomer hydrolase, was determined by constructing various deletion plasmids and inserting the lacUV5 promoter fragment of E. coli into the deletion plasmid. Two kinds of repeated sequences (RS-I and RS-II) were detected on pOAD2 by DNA-DNA hybridization experiments. These repeated sequences appeared five times (RS-I) or twice (RS-II) on pOAD2. One of the RS-II regions and the structural gene of the hydrolase overlapped.

Amidohydrolases↗

Natural relationship between bacteroides and flavobacteria.

Comparisons among 16S rRNA sequences from various eubacteria reveal a natural relationship between the bacteroides (represented by the Bacteroides fragilis sequence) and a phylogenetic unit that comprises the flavobacteria, cytophagae, flexibacteria, and others (represented by the Flavobacterium heparinum sequence). Although the relationship is not a close one, it is, nevertheless, specific. rRNAs from these two organisms are not only closer to one another in overall sequence than they are to outgroup species (such as Bacillus subtilis, Escherichia coli, Desulfovibrio desulfuricans, and Agrobacterium tumefaciens), but they show common idiosyncrasies (i.e., derived characteristics) in both rRNA sequences and higher-order structures.

Bacteroides↗

Low-molecular-weight thiols in streptomycetes and their potential role as antioxidants.

The intracellular low-molecular-weight thiols present in five gram-positive Streptomyces species and one Flavobacterium species were analyzed by high-performance liquid chromatography after fluorescence labeling with monobromobimane. Bacteria were chosen to include penicillin and cephalosporin beta-lactam producers and nonproducers. No significant amount of glutathione was found in any of the streptomycetes. Major intracellular thiols in all strains examined were cysteine, coenzyme A, sulfide, thiosulfate, and an unknown thiol designated U17. Those streptomycetes that make beta-lactam antibiotics also produce significant amounts of delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV), a key intermediate in their biosynthesis. In Streptomyces clavuligerus, a potent producer of beta-lactams, the level of ACV was low during the early phase of growth and increased rapidly toward the end of exponential growth, paralleling that of antibiotic production. These and other observations indicate that ACV does not function as a protective thiol in streptomycetes. U17 may have this role since it was the major thiol in all streptomycetes and appeared to occur at levels about 10-fold higher than those of the other thiols measured, including ACV. Purification and amino acid analysis of U17 indicated that it contains cysteine and an unusual amine that is not one of the common amino acids. This thiol is identical to an unknown thiol found previously in Micrococcus roseus and Streptomyces griseus. A high level of ergothioneine was found in Streptomyces lactamdurans, and several unidentified thiols were detected in this and other streptomycetes.

Amino Acids↗