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[Methanogenic sarcina from an anaerobic microbial community degrading p-toluene sulfonate].

The methanogenic strain MM isolated from an anaerobic microbial community degrading p-toluene sulfonate showed optimal values of temperature and pH for growth equal to 37 degrees C and 6.3-6.9, respectively. The doubling times of the isolate grown on methanol, acetate, and methylamines under the optimal conditions were 8.8, 19.1, and 10.3-28.1 h, respectively. The growth of strain MM was observed only when the cultivation medium contained casamino acids or p-toluene sulfonate. The G + C content of the DNA of the isolate was 40.3 mol%. This, together with DNA-DNA hybridization data, allowed the new isolate to be identified as a strain of the species Methanosarcina mazei. The new isolate differed from the known representatives of this species in that it was resistant to alkylbenzene sulfonates and able to demethylate p-toluene sulfonate when grown on acetate.

Acetates↗
PubMed1975

Investigations on the efficacy of surface disinfection and surface cleaning procedures. 1. Tests under real-life conditions.

The suitability of disinfection preparations is assessed on the basis of laboratory tests, different methods being used in the various countries. Since such model tests are rather inadequate when it comes to judging surface disinfectants, additional in-use tests are desirable. They might, in any case, serve as a reference system for judging the evaluation criteria which still differ widely at the moment. The experiments described in this study were chiefly designed to establish the effect of cleaning and disinfection measures on bacteria normally present on surfaces and on the artificial contamination of surfaces with Sarcinae as model germs. The tests were carried out in the halls on 5 floors of a medical (lift landings). "Rodac" plates were used to identify the germs. 3 disinfectants (aldehydes, phenol derivative), 3 disinfectant cleaning agents and soft soap were used. The preparations reduced the normal germ count by approx. 80 per cent. The reduction was mainly due to the cleaning effect (soft soap was as effective as the preparations with disinfectant properties). The effect on the "normal germ count" cannot, therefore, be used as sole criterion of disinfectant action. When the various preparations were applied in twice the concentration recommended for Staphylococcus hospitalism, the Sarcina count was reduced by 99 to 99.9 per cent within 2 hours. The efficacy of disinfectants and disinfectant cleaning agents was practically the same. Additional laboratory tests are necessary before the effect of soft soap can be finally assessed. In actual practice the unit count of pathogenic germs- such as Staphylococci and Klebsiellae- is too low to enable an objective assessment of a disinfectant to be made. On the other hand, artificial contamination with such pathogens is not possible because of the risk involved. The use of Sarcina lutea as test germ is therefore subjects to certain limitations. One of the prerequisites for using it is, for example, prior reduction of the normal germ count to values of less than 500/100 cm2. The second communication will report on investigations into the chemoresistance of Sarcina and how this compares with that of Staphylococcus aureus and Klebsiella. The need for such studies arose from our present investigation.

Cell Count↗
PubMed1978

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↗
PubMed1975

Investigations on the efficacy of surface disinfection and surface cleaning procedures. 2. Laboratory testing of the efficacy under conditions simulating those of real-life.

Using a standardised method (impression method by means of 'Rodac' plates and glass as well as ceramic surfaces as germ carriers) we tested for the efficacy of 3 surface disinfectants (aldehydes, aldehydes + detergent substances, phenol derivatives) and 3 disinfectant cleaning agents (aldehydes + detergent substances + wax) and 2 per cent soft soap solution on Staphylococcus aureus, Klebsiella pneumoniae and Sarcina lutea. The method enabled us to calculate the actual germ count reduction and also took account of the natural dying-rate of the test organisms. Their resistance to the different types of active substances varied greatly. Thus the 3 disinfectants had a better effect on Klebsiellae and Staphylococci than on Sarcinae. 2 of the 3 disinfectant cleaning agents, on the other hand, turned out to be less suitable for the elimination of Klebsiellae. It thus follows that Sarcina cannot be used as test organism for in-use tests (see Communication 1) unless the resistance of Staphylococci and Klebsiellae to the test preparation has first been established in laboratory experiments. The general question, namely whether in-use tests provide meaningful and accurate results remain to be elucidated. On the basis of our test results, the exclusive use of soap for cleaning floors, as a substitute of disinfection, should definitely be ruled out. Although soap reduces Gram-positive cocci well in actual practice, it is almost ineffective against problem germs such as Klebsiellae. What should be stipulated is the use of broad-spectrum disinfectants, i.e. they should, if possible, act equally well against different types of germs. From the commercially-available preparations that were tested, a phenol-base product gave the best results. On the basis of these results we venture to doubt that the use of aldehyde preparations which is customary at the moment, is the right approach to fight hospital infections.

Aldehydes↗
PubMed1975

Investigations on the efficacy of surface disinfection and surface cleaning procedures. 3. Evaluation of the results of in-use and laboratory tests.

3 disinfectants and 3 disinfectant cleaning agents were subjected to comparative tests as to their germicidal activity. These were (a) laboratory tests with germ carriers and (b) in-use tests with Sarcina-contaminated floor surfaces in a medical institute. The results, details of which are given in the two preceding publications, were assessed under the following angles: Validity of the test methods, requisite changes of the test procedures and practical usefulness of the different types of preparations. This assessment led to the following conclusions: New preparations should first be tested for their bacteriostatic activity according to DGHM and the neutralising agents should be determined. The suspensions tests - which remain to be standardised- provide information on the sensitivity to protein, hard water and detergent surface active substances. The germ carrier tests with operating-theatre tiles, carried out with at least 5 test germs, must result in a germ count reduction of more than 5 powers of ten (regardless of the elimination rate due to drying). In-use tests should stand at the end of the test series. Regular epidemiological studies are hardly feasible in all individual cases. An assessment on the basis of the behaviour of normal bacterial spores is likewise impracticable. The use of test germs cannot be dispensed with. Sarcina is a suitable species provided germ-carrier tests have first been carried out and have shown that Staphylococci, Klebsiellae and other high-risk germs are more sensitive to the preparation than Sarcina lutea. In the in-use tests, we consider a reduction by 2 to 3 powers of ten to be sufficient.

Cell Count↗
PubMed1994

16S ribosomal DNA sequences of anaerobic cocci and proposal of Ruminococcus hansenii comb. nov. and Ruminococcus productus comb. nov.

The 16S ribosomal DNA sequences of representative members of the family Peptococcaceae were determined. The members of the family examined were divided into the following four phylogenetic groups: Peptococcus niger ATCC 27731T (T = type strain), the Sarcina-Peptostreptococcus anaerobius group, the ruminococcus-coprococcus group, and the peptostreptococcus group. Peptococcus niger, the type species of the family, was not related to other members of the family. Peptostreptococcus anaerobius ATCC 27337T, the type strain of the type species of the genus Peptostreptococcus, was closely related to Clostridium sordellii NCIB 10717T (level of sequence similarity, 85%). Sarcina ventriculi GIFU 7886, a spore-forming anaerobic gram-positive coccus, clustered with Clostridium perfringens ATCC 13124T at a similarity value of 91%. Members of the Sarcina-Peptostreptococcus anaerobius group clustered with clostridia at similarity values ranging from 85 to 91%. The type strains of Peptostreptococcus prevotii, Peptostreptococcus asaccharolyticus, Peptostreptococcus micros, and Peptostreptococcus magnus clustered at levels of sequence homology of 84 to 93%. This cluster was not included in the Peptococcus niger group or the Peptostreptococcus anaerobius group. Thus, these members of the genus Peptostreptococcus should be separated from the other members of the genus and also from members of the family Peptococcaceae. The sequence of Peptostreptococcus productus ATCC 27340T was different from the sequences of Peptostreptococcus anaerobius and Peptococcus niger. The sequence of Streptococcus hansenii ATCC 27752T, a strictly anaerobic strain, was different from the sequences of other streptococci; this strain clustered with Peptostreptococcus productus, coprococci, and ruminococci. Several phenotypic characteristics of Streptococcus hansenii ATCC 27752T were similar to characteristics of ruminococci.(ABSTRACT TRUNCATED AT 250 WORDS)

Anaerobiosis↗
PubMed1968

Effect of gamma irradiation on the microflora of freshwater fish. 3. Spoilage patterns and extension of refrigerated storage life of yellow perch fillets irradiated to 0.1 and 0.2 megarad.

Maximal shelf life was determined and microbial flora were compared for irradiated (0.1 and 0.2 Mrad) and nonirradiated yellow perch fillets stored at 1 C. Shelf life was estimated by organoleptic determinations. Microbiological studies included determination of the effects of irradiation on the total aerobic microbial population, lag phase, and rate of growth. Genera of organisms isolated from fillets through the course of microbial spoilage were identified, and the proteolytic activity of the organisms was determined. Plate counts for fish prior to irradiation showed the presence of approximately 10(6) organisms per g of sample. Irradiation to 0.1 and 0.2 Mrad produced 1.4 and 3 logarithm reductions of the initial count, respectively. Irradiation to 0.1 and 0.2 Mrad approximately doubled the product's shelf life. Organisms initially isolated from the nonirradiated fillets, in order of decreasing number, consisted of Flavobacterium, Micrococcus-Sarcina, Achromobacter-Alcaligenes-Mima, Pseudomonas, Microbacterium, Vibrio, Bacillus, Corynebacterium, Lactobacillus, Brevibacterium, and Aeromonas. By the 6th and 9th days of fillet storage, Pseudomonas and the Achromobacter group were the predominant organisms. All members of the genus Flavobacterium, but not all members of the genus Pseudomonas, were proteolytically active on raw fish juice-agar and skim milk-agar media. The Achromobacter group was found to be nonproteolytic on both media. Residual flora of fillets irradiated to 0.1 and 0.2 Mrad consisted of the Achromobacter group, Lactobacillus, Micrococcus-Sarcina, and Bacillus. Their sequence in predominance, however, varied with dose level. Not all proteolytic bacteria in the fillets were eliminated by 0.1 and 0.2 Mrad; proteolytic Micrococcus-Sarcina survived these treatments.

Bacteria↗
PubMed2002

Screening of eight alkaloids and ten flavonoids isolated from four species of the genus Boronia (Rutaceae) for antimicrobial activities against seventeen clinical microbial strains.

Eight alkaloids and ten flavonoids isolated from four species of Boronia of the Rutaceae were screened against 17 clinical microbial strains. Of the test compounds, three acridone and one quinolone alkaloids and eight flavonoids were reported as novel natural products. Screening was carried out by the standard disc diffusion method. Of the tested compounds, six alkaloids and seven flavonoids including the novel products were active against six clinical strains. The active compounds showed mild to moderate activities against Bacillus subtilis, Staphylococcus aureus, Sarcina lutea, exterotoxigenic Escherichia coli, Salmonella typhi and Klebsiella sp. Of the active flavonoids, some exhibited fairly significant activity towards Staphylococcus aureus, Sarcina lutea, Salmonella typhi and Klebsiella sp. The flavonoids were observed to have higher spectrum and magnitude of activity than those of the alkaloids. A standard ampicillin disc was used to compare the results.

Alkaloids↗
PubMed1976

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↗
PubMed1965

Steroid lysis of protoplasts and effects of stabilizers and steroid antagonists.

Six synthetic antimicrobial steroids were examined for indications of their mechanism of action. Dequadin acetate, cetyl pyridinium chloride (CPC), and sodium deoxycholate were studied for comparison. Aerated cells of Sarcina lutea were washed, suspended in 1.06 M sucrose, and converted to protoplasts with 20 mug/ml of lysozyme. Lysis was measured optically at 650 mmu as a decrease in optical density. Screening tests with 50 mug/ml of each compound showed five steroids and CPC to be lytic. Protoplasts were strongly protected from lysis by pretreatment with 0.001 to 0.004 M spermine tetrahydrochloride. Other polyamines, such as spermidine phosphate, were less protective, and putrescine was ineffective. Uranyl nitrate (5 x 10(-4) M) rapidly agglutinated protoplasts and protected them from rupture by the lytic agents. Similar studies with 0.001 to 0.004 M Mg(++) showed varying degrees of protection, which, in most cases, was only temporary. Steroidal lysis did not appear to be related to chelation, since ethylenediaminetetraacetate did not cause lysis alone and antagonized some lytic compounds. Lecithin, Tween 80, Tween 20, and Span 20 at 0.05% exhibited certain effects on protoplast stability. Span 20 strongly prevented lysis by steroids. Tween 20 alone quickly caused protoplast rupture. Lecithin and Tween 80, which also caused lysis alone, interfered with lytic steroids and CPC. The test compounds were both inhibitory and lethal to cells of Sarcina lutea. The results suggest that direct action on cell membranes may be chiefly responsible for the antimicrobial properties of the steroids.

Bacteriolysis↗
PubMed1968

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↗
PubMed1970

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↗
PubMed1966

Deoxyribonucleic acid base composition of some members of the Micrococcaceae.

Auletta, Angela E. (Catholic University, Washington, D.C.), and E. R. Kennedy. Deoxyribonucleic acid base composition of some members of the Micrococcaceae. J. Bacteriol. 92:28-34. 1966.-Thirty-seven strains from the genera Micrococcus, Staphylococcus, Gaffkya, and Sarcina were examined for deoxyribonucleic acid base composition and biochemical activity. Organisms were tested for production of catalase, coagulase, deoxyribonuclease, oxidase, phosphatase, hydrogen sulfide, indole, and acetoin; nitrate reduction; gelatin, starch, and urea hydrolysis; citrate and ammonium phosphate utilization; NaCl tolerance; growth at 10 and 45 C, and growth in litmus milk. They were tested for production of acid from dextrose and mannitol under anaerobic conditions, and for aerobic production of acid from dextrose, mannitol, lactose, sucrose, raffinose, maltose, xylose, and glycerol. Organisms could be divided into two groups on the basis of guanine-cytosine (GC) content. Group I had an average GC content of 32%, and included all organisms which produced acid from dextrose. Group II had an average GC content of 62%, and included those organisms incapable of producing acid from dextrose under anaerobic conditions. Sarcina ureae had a GC content of 43%.

Carbohydrate Metabolism↗
PubMed2006

Variation in chemical composition and antibacterial activities of essential oils from two species of Houttuynia THUNB.

Houttuynia THUNB. (Saururaceae) has been used for dozens of years in China for the treatment of cough, leucorrhea and ureteritis. The essential oils from the two species: Houttuynia emeiensis and Houttuynia cordata sold in China under one trade name 'Yuxingcao', obtained by hydrodistillation, were analyzed by GC-MS. The results show that fifty-five components were identified and methyl nonyl ketone (2.10-40.36%), bornyl acetate (0.4-8.61%) and beta-myrcene (2.58-18.47%) were the most abundant components in oil, but the percentage of most of compounds in different species and parts varied greatly. The two fold broth dilution and agar dilution method were used to study essential oil of two Houttuynia THUNB. species for their antibacterial properties against microorganisms, Staphylococcus aureus and Sarcina ureae. The two fold dilution method was allowed to determine the minimum inhibitory concentration (MIC) of essential oil from different parts and species. Results showed that all essential oils possessed antibacterial effect, with MIC values in the range of 0.0625 x 10(-3) to 4.0 x 10(-3) ml/ml. However, essential oil from different parts and species differed clearly in their antibacterial activities. The essential oil from the aboveground part of the cultivated Houttuynia emeiensis exhibited higher activity than both parts of the wild and cultivated Houttuynia cordata when used on Staphylococcus aureus (MIC = 0.25 x 10(-3) ml/ml) and Sarcina ureae (MIC = 0.0625 x 10(-3) ml/ml), and had the same activity as the positive control ampicillin sodium.

Anti-Bacterial Agents↗
PubMed1984

The rate of evolutionary divergence of initiation factors IF2 and IF3 in various bacterial species determined quantitatively by immunoblotting.

Antibodies to Escherichia coli translational initiation factors IF2 and IF3 were used for an immunological comparison of unpurified proteins from the following genera: Salmonella, Serratia, Proteus, Aeromonas, Pseudomonas, Streptococcus, Sarcina and Bacillus. Immunological relatedness was compared by Ouchterlony double diffusion experiments and immunoblotting analysis. Immunoblotting is a quantitative technique for measuring levels of specific proteins in crude cell lysates. We have used this technique to measure immunological distance with the assumption that the levels of the various translational components are essentially the same in the different bacterial cells examined. Both immunodiffusion and immunoblotting analysis showed a similar evolutionary relationship between the various species for the two initiation factors examined: (Escherichia = Salmonella greater than Serratia greater than Proteus greater than Aeromonas greater than Pseudomonas). Little or no crossreactivity was found using either analysis with genera: Streptococcus, Sarcina and Bacillus. Using the immunoblot distance, the two initiation factors were shown to diverge at similar rates. One advantage the immunoblotting analysis has over other immunological techniques is that the antigens can be analyzed structurally. We found, for example, that the two forms of IF2 were present in all bacterial species which cross-reacted with anti-IF2, suggesting that both forms are functionally important. Because of its sensitivity, the immunoblot analysis may be more useful than other immunological techniques in studying species that are more distantly related.

Antibodies, Bacterial↗