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Comparative study of antibacterial activity of pancreatic juice in six mammalian species.

A comparative study of antibacterial activity of pancreatic juice was conducted on six mammalian species. Pancreatic juice collections were conducted as acute (rabbit, guinea pig, rat) and chronic (pig, sheep, cattle) experiments, in the former before and after stimulation [cholecystokinin (CCK) and secretin] and in the latter under basal conditions alone. Antibacterial activity was tested on Micrococcus pyogenes and compared with that of neomycin. The samples were tested under normal conditions and after heating and dilution. The pancreatic juice of rat showed no activity against Micrococcus pyogenes. The antibacterial activity of rabbit and guinea pig pancreatic juice under basal conditions was similar within the group but significantly higher than that of pig, sheep and cattle which also did not differ significantly within the group. On stimulation with CCK and secretin, no significant change could be observed in the potency of antimicrobial activity of pancreatic juice in the rabbit and guinea pig. The antibacterial activity remained unchanged after heating to 65 degrees C and upon dilution to 1:10.

Animals↗

Micrococci demonstrate a phosphatase activity which is repressed by phosphates and which can be differentiated from that of staphylococci.

The phosphatase activities of 114 micrococcal strains belonging to seven different species and of an additional 150 unspeciated micrococcal strains were evaluated on solid media at various pHs containing or not containing phosphates. In the presence of phosphates, only nine strains (five unspeciated strains, one Micrococcus luteus strain, and three Micrococcus varians strains) yielded a positive reaction on plates at pH 8. In media (at pH 8) deprived of phosphates, in contrast, all but 15 strains demonstrated clear-cut phosphatase activity. Acid phosphatase could not be evaluated on solid media since none of the strains grew satisfactorily on plates at pH 5. The phosphatase activities of seven (one or two for each species, which included phosphatase-negative strains) of the strains whose colonies proved phosphatase negative at pH 8 and of 18 (two or three strains per species) of those with phosphatase-positive colonies were evaluated at pH 5 and 8.5 in toluene-treated cells which had been grown in liquid media at pH 7 containing or not containing phosphates. All strains demonstrated distinct phosphatase activity at both pHs when grown in media not containing phosphates. In contrast, when strains were grown in the presence of such substances, virtually no activity was observed at pH 8.5, and, generally, a much reduced activity was observed at pH 5. The phosphatase activity of micrococci of the various species (three to eight strains per species) was also compared with that of staphylococci of different species (5 to 10 strains per species) by the methyl green-phenolphthalein diphosphate method, the sensitivity of which can be varied by using different enzyme substrates.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Phosphatase↗

Production of inhibitors of lytic activity in the Micrococcaceae.

Eight Staphylococcus xylosus strains lacking lytic activity (LA) were found to excrete agar-diffusible factors inhibiting the LA of LA-positive strains of the same species. The same eight strains, when tested against LA-positive indicator strains from other species of Micrococcaceae, caused marked LA inhibition only of strains of S, saprophyticus and S. cohnii, both species closely related to S. xylosus. Micrococci and planococci, unlike staphylococci, do not normally show LA, yet Micrococcus lylae and M. sedentarius strains produced inhibitors effective against the LA of almost all the indicator strains, and planococci inhibited the LA of a few indicator strains. Micrococcus luteus strains caused LA inhibition of homologous indicators only, i.e. exceptional M. luteus strains exhibiting LA. Thus, a specific and a non-specific type of LA inhibition can be distinguished, and it is suggested that the apparent lack of LA might, in some instances, be explained by the simultaneous production of a specific inhibitor.

Bacteriolysis↗

Phylogenetic and physiological diversity of Arthrobacter strains isolated from unconsolidated subsurface sediments.

Forty strains of Gram-positive, aerobic, heterotrophic bacteria isolated from saturated subsurface lacustrine, paleosol and fluvial sediments at the US Department of Energy's Hanford Site in south central Washington State were characterized by phylogenetic analysis of 16S rRNA gene sequences and by determination of selected morphological, physiological and biochemical traits. Phylogenetic analyses of 16S rDNA sequences from subsurface isolates in the context of similar sequences from previously described bacterial species indicated that 38 of the subsurface strains were most closely related to Arthrobacter: The other two strains appeared to be most closely related to Kocuria. The subsurface isolates fell into seven phylogenetically coherent and distinct clusters, indicating that there was a significant degree of diversity among them. Additional diversity was detected by analysis of cellular fatty acids and physiological traits. The general morphological, physiological and biochemical traits of the subsurface strains were consistent with those of Arthrobacter, Micrococcus and genera recently separated from Micrococcus, such as Kocuria. Some of the subsurface strains were phylogenetically closely related to certain species of Arthrobacter. (16S rDNA sequence similarities >99%). However, most of the subsurface isolates did not cluster with previously established species in phylogenetic analyses of 16S rRNA gene sequences or with hierarchical cluster analysis of cellular fatty acid profiles. Moreover, many of the subsurface isolates that were most closely related to Arthrobacter. also differed from all established species of that genus in several of their specific physiological characteristics. Most of the subsurface isolates, then, are likely to be novel strains or species of Arthrobacter.

Arthrobacter↗

The effect of variation of thermal processing on the microbial spoilage of chub-packed luncheon meat.

Process pasteurization values for reference temperature 70 degrees C (P70) were calculated from the temperature profiles of 250 g luncheon meat chubs cooked under experimental conditions. A simple equation relating Process P70-value and the time and temperature of cooking was derived. With minimal cooking (P70 = 40) the surviving microflora (10(3)/g) wad dominated by species of Lactobacillus, Brochothrix and Micrococcus. These organisms were destroyed by more intensive cooking (P70 = 105), leaving a flora (10(2)/g) composed of Bacillus and Micrococcus species. The spoilage that developed after 14 d storage at 25 degrees C reflected the severity of the heat treatment received by each chub: with P70 between 40 and 90, a Streptococcus spoilage sequence occurred; with P70 between 105 and 120, a Bacillus/Streptococcus spoilage sequence occurred; with P70 of 135 and above, a Bacillus spoilage sequence occurred. Cooking to a P70 = 75 was adequate to reduce the surviving microflora to the 10(2)/g level associated with current good manufacturing practice.

Bacillus↗

Scanning electron microscopy of dairy equipment surfaces contaminated by two milk-borne micro-organisms.

Ethanol dehydration followed by argon replacement induced drying (ARID) was found to be a suitable method for the preparation of glass, stainless steel and rubber surfaces which had been in contact with inoculated milk and which were to be examined using scanning electron microscopy (SEM). This technique was used to examine samples of all three materials which had been subjected to both single and repeated inoculation with whole milk containing a Pseudomonas sp. or a Micrococcus sp. and incubated for various periods. Some samples were also prepared for SEM using a cryofixation technique. The Pseudomonas sp. was found to proliferate on glass and stainless steel surfaces but not on rubber. Due to the clumping tendency of the Micrococcus sp. proliferation of this organism was more difficult to assess accurately. In general there was no difference in results obtained between single and repeated inoculation. Various factors which may have aided attachment of micro-organisms to surfaces were identified viz., surface channels present in stainless steel, milk deposits and the production of extracellular material. The value of using both the cryofixation and chemical preparatory techniques for the identification of artifacts is discussed.

Animals↗

Interaction of wheat-germ agglutinin with bacterial cells and cell-wall polymers.

Wheat germ agglutinin was found to agglutinate cells of Escherichia coli PAT 84, Micrococcus luteus, Staphylococcus aureus H, and of S. aureus 52A5, but not cells of S. aureus 52A2. Interaction of wheat germ agglutinin with a soluble linear peptidoglycan secreted by Micrococcus luteus and with the teichoic acid of S. aureus H was demonstrated by agar gel diffusion, quantitative precipitation and inhibition of hemagglutination of trypsinized rabbit erythrocytes. No interaction could be demonstrated with the teichoic acid from a phage-resistant mutant (S. aureus 52A2) which lacks A-acetyl-D-glucosamine residues. All interactions were specifically inhibited by low concentrations of chitotriose (GlcNAcbeta1 leads to 4GlcNAcbeta1 leads to 4GlcNAc) and the bacterial cell wall tetrasaccharide, GlcNAcbeta1 leads to 4MurNAcbeta1 leads to 4GlcNAcbeta1 leads to 4MurNAc. Hemagglutination-inhibition experiments showed that the linear peptidoglycan and the teichoic acid of S. aureus H were several thousand times more potent inhibitors of wheat germ agglutinin than was N-acetyl-D-glucosamine. Comparison of the efficiency of different saccharides in inhibition of hemagglutination and precipitation of polymers by wheat germ agglutinin, strongly suggests that secondary, non-specific interactions contribute to the binding of the lectin to the polymers.

Agglutination Tests↗

Hemolytic interactions of Dermatophilus congolensis.

The strains of Dermatophilus congolensis grew on blood agar with washed sheep erythrocytes with marked total hemolysis. In testing for hemolytic interactions they gave a significant synergistic effect of a characteristic shape with Rhodococcus equi and Streptococcus agalactiae, whereas with Staphylococcus aureus producing beta hemolysin and with Staphylococcus aureus producing delta hemolysin a simultaneous synergistic as well as antagonistic effect were observed. First of all a conspicuous inhibition of in the beta hemolysin zone began and then the hemolytic effect of D. congolensis was enhanced. A similar double reaction was also observed with Listeria ivanovii. With delta hemolysin there was an inhibition of the hemolytic effect of D. congolensis and at the same time a synergistic effect could be observed. Also D. congolensis gave a weak synergistic effect with Micrococcus lylae and Listeria monocytogenes, and a further weak antagonistic effect with alpha hemolysin of Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus chromogenes and Micrococcus luteus. No interaction of D. congolensis was established with Corynebacterium pseudotuberculosis.

Actinomycetales↗

Microbial uptake of lead.

Micrococcus luteus and Azotobacter sp. cells grown in broth in contact with a dialysis membrane containing lead bromide were found to immobilize 4.9 and 3.1 x 10(2) milligrams of lead per gram of whole cells, on a dry weight basis, respectively. Culture turbidity and cell count measurements on these and other cell cultures show that lead bromide, lead iodide, and lead bromochloride in concentrations approaching solubility limits have no detectable effect on overall growth rate and cell viability. Analyses of cellular subfractions reveal that fractions of cell wall plus membrane contain 99.3 and 99.1 percent of the lead found associated with Micrococcus luteus and Azotobacter sp., respectively. The remainder is found associated with the cytoplasmic fractions.

Azotobacter↗

Airborne bacteria in an urban environment.

Samples were taken at random intervals over a 2-year period from urban air and tested for viable bacteria. The number of bacteria in each sample was determined, and each organism isolated was identified by its morphological and biochemical characteristics. The number of bacteria found ranged from 0.013 to 1.88 organisms per liter of air sampled. Representatives of 19 different genera were found in 21 samples. The most frequently isolated organisms and their percent of occurence were Micrococcus (41%), Staphylococcus (11%), and Aerococcus (8%). The bacteria isolated were correlated with various weather and air pollution parameters using the Pearson product-moment correlation coefficient method. Statistically significant correlations were found between the number of viable bacteria isolated and the concentrations of nitric oxide (-0.45), nitrogen dioxide (+0.43), and suspended particulate pollutants (+0.56). Calculated individually, the total number of Micrococcus, Aerococcus, and Staphylococcus, number of rods, and number of cocci isolated showed negative correlations with nitric oxide and positive correlations with nitrogen dioxide and particulates. Statistically significant positive correlations were found between the total number of rods isolated and the concentration of nitrogen dioxide (+0.54) and the percent relative humidity (+0.43). The other parameters tested, sulfur dioxide, hydrocarbons, and temperature, showed no significant correlations.

Air Microbiology↗

Science of hemolytic activity of some radiation-resistant micrococci in food.

Micrococci resistant to 1 Mrad of gamma radiation were isolated from irradiated chicken. Three isolates were hemolytic on blood agar plates and were selected for further study. Two other radiation-resistant micrococci, Micrococcus radiodurans and Micrococcus radiophilus, were included in the study because there is only a very limited amount of information regarding hemolytic activity of these organisms and their potential role of public health importance. Tests to determine hemolytic patterns, hemolytic activity of extracellular substances, leukocytic activity, presence of enzymes commonly associated with pathogenicity (coagulase, deoxyribonuclease, phosphatase), and pathogenicity for laboratory animals all suggested that the organisms would not be of public health significance.

Agar↗

Effects of nitric oxide and nitrogen dioxide on bacterial growth.

The effects of low concentrations of nitric oxide (NO) and nitrogen dioxide (NO2) on actively dividing cultures of Staphylococcus aureus, Micrococcus luteus, Micrococcus roseus, Serratia marcescens, Bacillus subtilis, Bacillus circulans, Bacillus megaterium, and Bacillus cereus were studied. Fresh cultures of each organism were incubated for 24 h at 25 degrees C on both nutrient agar and mineral salts glucose agar plates under atmospheres containing various low concentrations of NO in air (0 to 1.9 ppm [0 to 2.0 micrograms/g of air]), NO2 in air (0 to 5.5 ppm [0 to 8.8 micrograms/g of air]), or NO and NO2 in air. Bacteria grown under air only were used as controls. After incubation, the colonies that developed on the plates were counted. None of the bacteria tested was affected by NO or NO2 at the indicated concentrations while growing on nutrient agar. Serratia marcescens, B. circulans, B. subtilis, B. megaterium, and B. cereus grown on mineral salts glucose agar were not significantly affected by NO or NO2. Low concentrations (0 to 1.9 ppm) of NO were bacteriostatic to log-phase cultures of M. roseus, M. luteus, and Staphylococcus aureus grown on mineral salts glucose agar. Bacteriostatic activity over a 24-h interval was maximal at an initial NO concentration of 1 ppm. Appreciable amounts of NO2 were produced in 24 h at initial NO concentrations greater than 1 ppm. These results suggest that NO2 may reduce the bacteriostatic activity of NO. Low concentrations (0 to 5.5 ppm) of NO2 in air did not affect any of the bacteria tested. At these low concentrations, NO affected bacterial growth, although NO2, NO2-, and NO3- did not. In addition, it was determined that the bacteriostatic activity observed in this study was not due to an increase in the acidity of the medium.

Bacillus↗

Effect of physiological age on radiation resistance of some bacteria that are highly radiation resistant.

Physiological age-dependent variation in radiation resistance was studied for three bacteria that are highly radiation resistant: Micrococcus radiodurans, Micrococcus sp. isolate C-3, and Moraxella sp. isolate 4. Stationary-phase cultures of M. radiodurans and isolate C-3 were much more resistant to gamma radiation than were log-phase cultures. This pattern of relative resistance was reversed for isolate 4. Resistance of isolate 4 to UV light was also greater during log phase, although heat resistance and NaCl tolerance after heat stress were greater during stationary phase. Radiation-induced injury of isolate 4 compared with injury of Escherichia coli B suggested that the injury process, as well as the lethal process, was affected by growth phase. The hypothesis that growth rate affects radiation resistance was tested, and results were interpreted in light of the probable confounding effect of methods used to alter growth rates of bacteria. These results indicate that dose-response experiments should be designed to measure survival during the most resistant growth phase of the organism under study. This timing is particularly important when extrapolations of survival results might be made to potential irradiation processes for foods.

Acinetobacter↗

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↗

Nuclease production and lysostaphin susceptibility of Staphylococcus aureus and other catalase-positive cocci.

Some strains of Staphylococcus epidermidis and Micrococcus sp. produce nucleases. However, thermal stability was shown to be unique to the nucleases of S. aureus. In addition, two micromethods for susceptibility testing to lysostaphin were more precise and convenient than anaerobic glucose fermentation in distinguishing between the genera Staphylococcus and Micrococcus.

Anti-Bacterial Agents↗

Amino acid sequence of the murein of Planococcus and other Micrococcaceae.

The amino acid composition and amino acid sequence of the murein (peptidoglycan) of 10 strains of planococci were studied. It is shown that the peptide subunit consists of muramyl-l-alanyl-gamma-d-glutamyl-l-lysyl-d -alanine. The cross-linking of two adjacent peptide subunits is mediated by d-glutamic acid which is bound to the epsilon-amino group of lysine by its gamma-carboxyl group and to the carboxyl group of d-alanine of an adjacent peptide subunit by its amino group. About 20 to 25% of the peptide subunits are not cross-linked. The murein structure of the different species and strains of Micrococcus, Staphylococcus, and Sarcina are compared. It is evident that the murein structure is a very good criterion for grouping the micrococci. In addition, some of these groups are fairly well defined by physiological properties as well as by their guanine + cytosine content of the deoxyribonucleic acid e.g., Micrococcus, Staphylococcus, Planococcus, Sarcina ureae. Other groups, represented by a single or a few strains only, such as M. varians NTCC 7281, M. radiodurans, M. freudenreichii ATCC 407, and M. luteus ATCC 398, need further investigation.

Amino Acid Sequence↗

Use of antibody to membrane adenosine triphosphatase in the study of bacterial relatioships.

An antiserum to Ca(2+)-activated adenosine triphosphatase from membranes of Micrococcus lysodeikticus cross-reacted in agar gels with membrane adenosine triphosphatases from other pigmented micrococci and related species. Species of Micrococcus and Sarcina showed different levels of inhibition of adenosine triphosphatase activities in heterologous reactions with antiserum. Inter- and intraspecific relationships based on the inhibition reaction were compared with an independent parameter, namely the quantitative and qualitative composition of the bacterial membrane phospholipids and fatty acids. The guanine plus cytosine contents in the deoxyribonucleic acid of the species studied correlated well with the serological cross-reactivity of adenosine triphosphatases from their membranes. The types of cross-bridges found in the peptidoglycans of these cocci were also compared with the other properties. The results suggest that an antiserum specific for a major membrane protein may be a reliable and most useful adjunct in studying bacterial serotaxonomy.

Adenosine Triphosphatases↗

In vitro synthesis of the unit that links teichoic acid to peptidoglycan.

The role of cytidine diphosphate (CDP)-glycerol in gram-positive bacteria whose walls lack poly(glycerol phosphate) was investigated. Membrane preparations from Staphylococcus aureus H, Bacillus subtilis W23, and Micrococcus sp. 2102 catalyzed the incorporation of glycerol phosphate residues from radioactive CDP-glycerol into a water-soluble polymer. In toluenized cells of Micrococcus sp. 2102, some of this product became linked to the wall. In each case, maximum incorporation of glycerol phosphate residues required the presence of the nucleotide precursors of wall teichoic acid and of uridine diphosphate-N-acetylglucosamine. In membrane preparations capable of synthesizing peptidoglycan, vancomycin caused a decrease in the incorporation of isotope from CDP-glycerol into polymer. Synthesis of the poly (glycerol phosphate) unit thus depended at an early stage on the concomitant synthesis of wall teichoic acid and later on the synthesis of peptidoglycan. It is concluded that CDP-glycerol is the biosynthetic precursor of the tri(glycerol phosphate) linkage unit between teichoic acid and peptidoglycan that has recently been characterized in S. aureus H.

Bacillus subtilis↗