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A separation of staphylococci and micrococci based on serological reactivity with antiserum specific for polyglycerophosphate.

A serological reaction with the antiserum against heterophile polyglycerophosphate (PGP) was evaluated for genus level differentiation among strains of Staphylococcus and Micrococcus-Sarcina spp.. Hot saline extracts from whole cells of Staphylococcus spp. strongly reacted with the PGP antiserum, whereas those of Micrococcus-Sarcina spp. did not. Likewise, phenol-water extracts from whole cells of Micrococcus-Sarcina spp. were not reactive with the PGP antiserum, although the extracts of staphylococcal cells again gave a strong reaction with the antiserum. This study indicates that extracts from Micrococcus-Sarcina spp. have no antigen reactive with the PGP antiserum and can thus be differentiated from extracts of Staphylococcus spp. which react strongly with the PGP antiserum.

Antibodies, Bacterial↗

Mitochondrial DNA sequences reveal extensive cryptic diversity within a western American springsnail.

We analysed cytochrome c oxidase subunit I and NADH dehydrogenase subunit I sequence variation among 29 populations of a widely ranging southwestern springsnail (Pyrgulopsis micrococcus) and 18 regional congeners. Cladistic analyses of these sequences depict P. micrococcus as a polyphyletic composite of five well-supported clades. Sequence divergences among these clades and subclades imply the possible occurrence of as many as seven or eight cryptic species in addition to P. micrococcus. Our finding that P. micrococcus contains multiple, genetically distinct and geographically restricted lineages suggests that diversification within this highly speciose aquatic genus has been structured in large part by the operation of terrestrial barriers to gene flow. However, these sequence data also indicate that recent dispersal among hydrographically separated areas has occurred within one of these lineages, which we attribute to passive transport on migratory waterbirds.

Animals↗

Bacterial populations associated with the dirty area of a South African poultry abattoir.

Bacterial populations associated with three sample types from the neck region of poultry carcasses in the dirty area of an abattoir were characterized. Sample types before and after scalding were skin only, feathers only, and a skin and feather combination. The neck skin of carcasses after the defeathering processing stage was also sampled. Bacterial populations associated with water from the scald tank, rubber fingers at the exit of the defeathering machine, and air in the dirty area were also characterized. Bacterial colonies (751) were randomly isolated from yeast extract-supplemented tryptone soya agar plates exhibiting 30 to 300 colonies. Micrococcus spp. were isolated in the highest proportion from pre-and postscalded carcass samples (63.5 to 86.1% of isolates), regardless of the sample type. Conversely, Enterobacteriaceae (40.3%), Acinetobacter (19.4%), and Aeromonas/Vibrio (12.5%) species predominated on neck skin samples taken from mechanically defeathered carcasses. Isolates from the rubber fingers were, however, predominantly Micrococcus spp. (94.4%). Bacterial groups isolated in the highest proportion from scald tank water samples were Micrococcus spp. (38.3%), species of Enterobacteriaceae (29.1%), and lactic acid bacteria (17.0%). Corynebacterium spp., species of Enterobacteriaceae, and Micrococcus spp. were dominant on air settle plates.

Abattoirs↗

Quantitative and qualitative aspects of bacterial flora of Karachi coastal water shrimp (Penaeus merguiensis and Metapenaeus monoceros).

Bacterial counts were made over a period of two years of two important commercial shrimp varieties of Karachi coastal waters. Bacteria were also isolated and identified. Total number of bacteria were found to be remarkably equal at 37 degrees, 30 degrees and 25 degrees C. Aerobic plate count of Penaeus merguiensis at 37 degrees C ranged from 1.2 X 10(5) to 6.0 X 10(7) CFU/g (Median 1.8 X 10(6) CFU/g), and were predominantly Vibrio, Micrococcus, Pseudomonas, Staphylococcus, Bacillus, and Flavobacterium. The corresponding count at 30 degrees C ranged from 3.2 X 10(5) to 4.7 X 10(7) CFU/g (Median 2.6 X 10(6) CFU/g). The bacterial flora in order of predominance were Vibrio, Micrococcus, Pseudomonas, Moraxella, Flavobacterium, Bacillus, Alteromonas, and Acinetobacter. The 25 degrees C counts ranged from 5.3 X 10(5) to 8.5 X 10(7) CFU/g (Median 3.1 X 10(6) CFU/g), the flora was composed of Vibrio, Moraxella, Micrococcus, Pseudomonas, Flavobacterium, Bacillus, Alteromonas, and Acinetobacter in order of predominance. The aerobic plate count of Metapenaeus monoceros at 30 degrees C ranged from 8.4 x 10(5) to 3.8 x 10(7) CFU/g (Median 2.9 x 10(6) CFU/g). The bacterial flora in order of predominance were Vibrio, Micrococcus, Moraxella, Pseudomonas, Alteromonas, Flavobacterium and Staphylococcus. No significant qualitative or quantitative difference was obtained between the two shrimp species. The presence of Staphylococcus at 37 degrees C was attributed to favourable incubation temperature as well as to excessive unsanitary handling while the absence of Moraxella and Alteromonas putrefaciens at this temperature was considered due to the psychotrophic nature of these organisms.

Animals↗

A membrane-associated lipomannan in micrococci.

Membranes of Micrococcus lysodeikticus, Micrococcus flavus and Micrococcus sodonensis contain acidic lipomannans. Lipoteichoic acids could not be detected in these organisms, and the suggestion that they are substituted for by the lipomannans is strengthened by the chemical and physical resemblances between the two polymers. The mannans contain glycerol, ester-linked fatty acids and mono-esterified succinic acid residues, giving them both hydrophobic and charged properties. The M. lysodeikticus mannan has a chain of about 60 hexose units with two branch points, and is joined at its reducing end to the 1-position of a glycerol moiety bearing two fatty acid residues. Succinic acid on the mannan enables it to bind Mg2+ efficiently, and the polymer is firmly associated with the cytoplasmic membrane, probably by intercalation of its fatty acids with those of the membrane lipids.

Borohydrides↗

Classification of micrococci on the basis of deoxyribonucleic acid homology.

The DNA homology relationships of 25 micrococci (15 strains of Micrococcus, eight strains of Sarcina and two strains of Staphylococcus) were studied by the deoxyribonucleic acid hybridization method using nuclease S1, an endonuclease specific for single-stranded DNA molecules. Nineteen of the strains were classified into three groups. Group I contained Micrococcus lysodeikticus IAMI056, M. luteus IAMI1010, M. flavus IAMI2005 and IAMI2006, Sarcina flava IAMI2007 and IAMI1006. S. subflava IAMI2009, S. lutea ATCC381, and ATCC382, and M. luteus IAMI1006. Group II contained M. roseus IAMI315, ATCC412, ATCC185 and IAMI295. Group III contained S. lutea IAMI099, IFO3232 and ATCC383, M. varians ATCC399 and Staphylococcus lactis ATCC15306. Micrococcus luteus IAMI097, M. varians ATCC19099 and ATCC19100, M. conglomeratus IAMI459 and IAMI470, and St. aureus IAMI011 could not be assigned to any of the three groups. The grouping corresponds to that derived from the results of differential lysis by lysozyme, 'lytic enzyme 2' from Cytophaga sp., or Streptomyces albus G enzyme; and to types of peptidoglycan in the cell walls and genetic transformation. The usefulness of classification based on sensitivity to various lytic enzymes was demonstrated. Group I probably coincides with M. luteus of Bergey's Manual of Determinative Bacteriology (1974), and groups II and III with M. roseus and M. varians respectively.

DNA, Bacterial↗

Temporal study of the staphylococci and micrococci of normal infant skin.

Staphylococcus and Micrococcus populations were collected from the healthy skin of 10 infant subjects. Infants were sampled from 1 day to 32 weeks of age. Species were characterized by approximately 30 different morphological, physiological, and biochemical characters. Staphylococci were the predominant inhabitants of normal skin, whereas micrococci were found only occasionally in this environment. Staphylococcus epidermidid, S. haemolyticus, and S. hominis were the predominant and persistent staphylococci. These species constituted a high percentage of the total aerobic bacterial flora of infant skin. Micrococcus luteus and M. kristinae were the prevalent micrococci found on infant skin. Only limited correlation between Staphyloccus and Micrococcus populations and infant age or body area sampled was indicated by this study.

Age Factors↗

Identification of Micrococcaceae in clinical bacteriology.

The cellular morphology, identifying physiological characteristics, and a key to the human genera of Micrococcaceae are presented with flow charts for identification of aerobic and anaerobic isolates. These flow charts can be amended as desired, depending upon the degree of accuracy desired. Micrococcaceae isolates in a 350-bed private general hospital during a 15-week period are tabulated to show relative numbers of the different genera and species, with their probable relationship to infection or contamination. Only 11 of the 220 Micrococcaceae isolates were not Staphylococcus; no Sarcina or Peptococcus were isolated. Of the Staphylococcus isolates, 61% were S. epidermidis. Almost 18% of the S. aureus isolates were coagulase-negative. Of the S. aureus isolates, 80% of the coagulase-positive isolates were infecting agents, as were 67% of the coagulase-negative S. aureus isolates, compared to only 48% of S. epidermidis isolates. Two of four Gaffkya isolates but only one of seven Micrococcus isolates were infecting agents. If coagulase production is used as the sole criterion for speciation of staphylococci, and Micrococcus is not differentiated from Staphylococcus, the term "coagulase-negative staphylococci" does not differentiate three distinct levels of pathogenicity. Coagulase-negative S. aureus is more virulent than S. epidermidis or Gaffkya, which are more virulent than Micrococcus or Sarcina.

Bacteriological Techniques↗

Use of lysostaphin in the isolation of highly polymerized deoxyribonucleic acid and in the taxonomy of aerobic Micrococcaceae.

By use of the staphylolytic enzyme lysostaphin, a method was devised for isolating and purifying highly polymerized deoxyribonucleic acid (DNA) from lysostaphinsusceptible Micrococcaceae. Staphylococcus aureus DNA isolated by this procedure gave an estimated molecular weight of ca. 2 x 10(8) and a residual protein content of 2.3%. The mole percentage of guanine + cytosine (GC) present in the DNA from 21 strains of aerobic Micrococceae was determined by buoyant density in cesium chloride. DNA from 12 biochemically typical members of the genus Staphylococcus gave a mean GC composition of 35.2 +/- 0.5 mole per cent. Four biochemically atypical Staphylococcus strains and one biochemically typical strain of the genus Micrococcus (M. candicans) were found to be susceptible to lysostaphin and gave typical Staphylococcus spp. GC base ratios. One biochemically atypical member of the genus Micrococcus (M. varians) was not susceptible to lysostaphin and gave a typical Micrococcus spp. GC base ratio. Lysostaphin susceptibility is an easy test to perform, and the results of this test appear to correlate with GC base ratio studies of the genera of Micrococcaceae.

Anti-Bacterial Agents↗

A serological method for distinguishing coagulase-negative staphylococci from micrococci.

Absorbed antisera prepared against heat-killed suspensions of a coagulase-negative staphylococcus and of a micrococcus were used in a slide-agglutination test. A collection of 176 coagulase-negative aerobic cocci, from which Staphylococcus aureus had been excluded, was classified into staphylococci and micrococci by means of the test for anaerobic utilization of glucose. Of 113 staphylococci, 99 reacted with the anti-staphylococcus serum, seven with the micrococcus serum, and seven with neither; of 63 micrococci, 55 reacted with the micrococcus serum and eight were untypable. Thus 161 of 176 strains (91%) were classifiable serologically, and in 154 of 161 instances (95%) the results of the serological and biochemical classification were in agreement.

Absorption↗

[Biosorption of heavy metals by bacteria isolated from activated sludge].

In this paper, biosorption efficiency of heavy metals by bacteria isolated from activated sludge was tested. It was shown that Pseudomonas pseudoalcaligenes and Micrococcus luteus GC subgroup B had high biosorption capacities for Cu(II) and Pb(II) ions. The adsorption of the two ions on Pseudomonas pseudoalcaligenes and Micrococcus luteus GC subgroup B exhibited Langmuir adsorption behavior with regression coefficient of 0.99. The pH value was the most important effect factor on biosorption of metal ions. The biosorption kinetic curves of Cu(II) and Pb(II) ions by cells showed two processes: rapid sorption on cell surface and slowly longer-term transformation into cells. Cu(II) ion can be desorbed effectively from cells of Micrococcus luteus GC subgroup B.

Absorption↗

Bacterial biosorbent for removing and recovering copper from electroplating effluents.

Investigations were carried out to study the removal and recovery of Cu(II) ions from wastewater by Micrococcus sp. The Langmuir isotherm model described very well the equilibrium behavior of copper biosorption, with maximum biosorption capacity (q(max)) reaching 52.1 mg Cu2+/g dry cell at pH 6. Biomass prewashed with sulfuric acid (0.05 mol 1(-1)) and sodium sulfate (1 mol l(-1)) solutions were shown to increase the copper removal capabilities up to 27% and 16%, respectively. Copper uptake by cells was negligible at pH 2.0 and then increased quickly with increasing pH until 6.0. Cells of Micrococcus sp. were immobilized in 2% calcium alginate and 10% polyacrylamide gel beads. A counter-current process comprising a series of immobilized cell reactors was developed for removing and recovering copper from electroplating effluents. This process was capable of producing an effluent at low copper concentration, with only a minimum amount of desorbing agent used. The technique of scanning electron microscopy coupled with X-ray dispersion analysis shows that Cu2+ exchanged with K+ and Ca2+ on the cell wall of Micrococcus sp., thereby suggesting ion exchange as one of the dominant mechanisms of metal biosorption for this bacterial strain.

Adsorption↗

[Analysis of methods for optimizing lysozyme determination].

In order to optimize the detection of lysozyme in the clinic, studies on photometric and agar diffusion methods were performed with different preparations of micrococcus luteus (lysodeikticus). Living or living lyophilized preparations of micrococcus were suitable. The influence of temperature, test period and lysozyme concentration on the result was demonstrated. The lysoplate technique has several advantages in clinical routine tests, however an exact adjustment of the reagents and the test is necessary (living micrococcus, 3 hours incubation, 50 degrees C test temperature). The lysozyme measurements of different papers are transferable not without reserve and require a careful interpretation in respect of reproducibility and clinical relevance.

Animals↗

[Taxonomic position of the genus Mycococcus].

An attempt to elucidate the taxonomic position of the genera Mycococcus has been made. For this purpose the full range of the taxonomically important properties were investigated among microbial cultures preliminary identified as Mycococci. As a result of comparison Mycococcus with Micrococcus the taxonomic status of the genus Mycococcus was no longer confirmed. On the grounds of the published data and the author's experimental results it was concluded that genus Micrococcus was closely related to genus Arthrobacter. The possibility to place the genus Micrococcus within the group of coryneform bacteria is being discussed.

Actinomycetales↗

[Bacterial reducing qualities of copper-containing and non-copper-containing materials. I. Contamination and sedimentation in humid and dry conditions].

In this examination the bactericide qualities of different materials were tested. Particularly the effect of humidity was examined. 3 materials containing copper ("copper", "brass" and "brass sheet") and 2 materials not containing copper ("polyamide B" and "high-grade steel") were tested. The surfaces of the 5 types of material were contaminated with a suspension of 2 different bacteria species (Staphylococcus aureus and Micrococcus luteus). After defined exposure times the survival rate of the microorganisms on the surfaces was determined by the standardized method of the "Guidelines of the German Society of Hygiene and Microbiology". Additionally the sedimentation of airborne microorganisms on the materials was tested by determining the number of colony-forming-units after defined exposure times. In further examinations the materials were contaminated by hands, which were dipped into a suspension of different microorganisms (Micrococcus luteus, Staphylococcus aureus, Enterococcus faecium, Escherichia coli, Enterobacter cloacae and Pseudomonas aeruginosa). The following ways of contaminating the test materials were chosen: single contamination with wet hands, repeated contamination with wet and dry hands. After defined exposure times the survival rate of the microorganisms on the surfaces was determined. The results show, a bactericide quality of copper depends on the percent of copper-content of the tested materials, exposure time, humidity, frequency of contamination and bacteria species. Humidity and only single contamination cause a reduction of bacteria. Reduction of gram-negative bacteria is more easily than reduction of gram-positive bacteria. Enterococcus faecium shows the highest, Pseudomonas aeruginosa the lowest reduction rate of bacteria in the following sequence: Enterococcus faecium, Escherichia coli, Enterobacter cloacae, Micrococcus luteus, Staphylococcus aureus and Pseudomonas aeruginosa.

Air Microbiology↗

Cloning and expression of a gene encoding Moraxella sp. CK-1 autolysin.

A gene of Moraxella sp. CK-1 encoding cell wall lytic activity has been cloned and expressed in E. coli. A genomic library of Moraxella sp. CK-1 was constructed in the multifunctional phagemid vector pT7T3 19u, with partial Sau3A digests of Moraxella sp. CK-1 genomic DNA inserted at the BamHI restriction site. Screening of about 5,000 transformants for cell lysis activity in LB agar plates containing Micrococcus Iuteus cells gave one positive clone harboring the 3.7 kb insert (pMXA282). Restriction mapping and deletion analysis of the recombinant plasmid carrying a 3.7 kb insert suggested that the autolysin gene was located within a 1.1 kb BamHI-PstI fragment. Analysis of extracts of E. coli clone harboring recombinant plasmids on renaturing SDS-polyacrylamide gels containing heat-killed Micrococcus luteus cells showed a clear zone around a polypeptide of about 32 kDa. Lytic activity against Micrococcus luteus cell walls by the cloned autolysin was maximum at pH 9.0. Even in conditions of over pH 10.0, this cloned autolysin showed a vigrous lytic activity. Southern blot analysis suggested the existence of other homologous regions in Moraxella sp. CK-1 genome.

Bacteriolysis↗

Insecticides: their effect on microorganisms and persistence in rice soil.

A field experiment was conducted to investigate the effect of four insecticides, HCH, phorate, carbofuran and fenvalerate, at recommended doses on the preponderance of bacteria, actinomycetes and fungi. We also measured the persistence of the insecticides in the rhizosphere soil of rice. HCH and fenvalerate stimulated the proliferation of all of the microorganisms significantly. Phorate increased the population of bacteria and actinomycetes. Carbofuran accentuated the preponderance of actinomycetes in soil. Insecticides, in general, did not have marked influence on the proliferation of Bacillus, Streptomyces, Aspergillus and Fusarium in soil. However, we observed a stimulation of growth of Staphylococcus, Proteus and Sarcina with HCH, Pseudomonas, Corynebacterium, Erysipelothrix and Rhizopus with phorate, Serratia, Corynebacterium, Klebsiella, Escherichia, Rhizopus and Humicola with carbofuran, and Staphylococcus, Sarcina, Klebsiella and Nocardia with fenvalerate. On the other hand, there was an inhibition in growth of Pseudomonas, Micrococcus, Nocardia and Penicillium with HCH, of Pseudomonas, Micrococcus and Penicillium with carbofuran, and of Pseudomonas, Micrococcus and Micromonospora with fenvalerate. Different types of insecticides exhibited differential patterns of dissipation in soil. HCH had the highest persistence followed by phorate, carbofuran and fenvalerate, respectively.

Actinomycetales↗

Citricoccus muralis gen. nov., sp. nov., a novel actinobacterium isolated from a medieval wall painting.

A Gram-positive, aerobic, spherical actinobacterium, designated strain 4-0(T), was isolated from a medieval wall painting and characterized to determine its taxonomic position. The peptidoglycan of strain 4-0(T) was of type A4alpha, with lysine as the diagnostic cell wall diamino acid and an interpeptide bridge of Lys-Gly-Glu. Its quinone system contained predominantly MK-9(H2) (64%) and its polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, four unknown glycolipids, two unknown phospholipids and an unknown lipid. The fatty acid profile of strain 4-0(T) was represented by significant amounts of ai-C15:0 and moderate amounts of ai-C17:0, i-C16:0 and i-C15:0 fatty acids. Spermidine was predominant in the polyamine pattern. The G+C content of the genomic DNA was 68 mol%. Comparative 16S rDNA sequence studies revealed the highest similarity values (95.4-96.1%) between strain 4-0(T) and species of the genus Micrococcus and certain species of the genus Arthrobacter, including Arthrobacter pascens DSM 20545(T), Arthrobacter ramosus DSM 20546(T), Arthrobacterprotophormiae DSM 20168(T), Arthrobacternicotianae DSM 20123(T) and Arthrobacter globiformis DSM 20124(T). Phylogenetic analyses demonstrated that strain 4-0(T) branches deeply on the Micrococcus lineage. Because it is almost equidistant phylogenetically from the genera Micrococcus and Arthrobacter and possesses significant differences in chemotaxonomic characteristics from members of these genera, it is suggested that strain 4-0(T) be classified as a novel species in a new genus; the name Citricoccus muralis gen. nov., sp. nov. is proposed. The type strain is strain 4-0(T) (= DSM 11442(T) CCM 4981(T)).

Austria↗