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PubMed1975

Systematic difference in the methylation of ribosomal ribonucleic acid from gram-positive and gram-negative bacteria.

A survey of gram-positive and gram-negative organisms was performed to compare the distributionof N6-methylated adenine. It was found that (i) all the gram-positive strains tested, Staphylococcus aureus, Sarcina lutea, Bacillus stearothermophilus, Bacillus subtilis, and Bacillus megaterium, contain neither N6-monomethyl adenine (m6A) nor N6-dimethyladenine (m26A) in 23S ribosomal ribonucleic acid (rRNA). In the case of S. aureus and Streptococcus pyogenes, strains which are clinically resistant to erythromycin contain m26A. (ii) The gram-negative strains Salmonella typhimurium, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, and Klebsiella pneumoniae all contain m6A but not m23A in 23S rRNA. These observations suggest the existence of at least one systematic structural difference between the ribosomes of the two classes of bacteria. Because of the demonstrated relationship between N6-dimethylation of adenine in 23S rRNA and clinical resistance to macrolide, lincosamide, and streptogramin B-type antibiotics in staphylococci and streptococci, the observed systematic differences found in rRNA methylation combined with greater cellular permeability may be related to the relatively greater efficacy of macrolide, lincosamide, and streptogramin B-type antibiotics in treating infections caused by gram-positive organisms.

Adenine↗
PubMed1967

Effect of azasteroids on gram-positive bacteria.

A group of nitrogen-containing steroids closely related in structure was screened for antibacterial activity, by use of Bacillus subtilis and Sarcina lutea as the test organisms. The most active compounds were cholesterol derivatives containing a tertiary or quaternary nitrogen in, or attached to, the A ring. Similar methyltestosterone or progesterone derivatives were inactive. All of the cholesterol derivatives that inhibited growth were surfactant, and, structurally, they would be classified as cationic detergents. Some of the inactive compounds were surfactant, but, structurally, they would be classified as nonionic detergents. Certain features of the antibacterial activity of one of the active steroids-ND 212 (4-dimethylaminoethyl-4-aza-5-cholesten-3-one methiodide)-were studied. Growth of a culture of B. subtilis containing 5 x 10(7) cells per milliliter was inhibited by 1 mug/ml (1.7 x 10(-6)m) of ND 212. The amount of growth inhibition was directly related to both cell and steroid concentration. Loss of viability was rapid and irreversible. With B. subtilis, cell lysis was observed. With S. lutea grown in C(14)-glucose, ND 212 caused release into the media of up to 25% of the cellular radioactivity. Extensive leakage occurred before loss of viability was observed. At bacteriostatic azasteroid concentrations, there was little leakage. ND 212 was readily bound in large amounts to B. subtilis cells. Inactive azasteroids were bound poorly. C(14)-cholestanone was also bound, whereas C(14)-methyltestosterone and C(14)-progesterone were not bound in significant amounts. At least 50% of the bound C(14)-cholestanone was associated with the membrane fraction.

Bacillus subtilis↗
PubMed1968

Lytic effects of staphylococcal alpha-toxin and delta-hemolysin.

Several preparations of staphylococcal alpha-toxin and delta-lysin were studied in order to compare hemolytic activity with capacity to lyse bacterial protoplasts. delta-Lysin in relatively low concentration lysed protoplasts of Sarcina lutea, protoplasts of Streptococcus faecalis, and spheroplasts of Escherichia coli. Lysis of bacterial protoplasts by preparations of alpha-toxin appeared to be due to contamination of the preparations with delta-lysin. Data comparing the protoplast-lysing activity of various lytic agents are presented.

Blood↗
PubMed1980

Comparison of methods available for assay of chloramphenicol in clinical specimens.

Eight methods for the assay of chloramphenicol in clinical samples were compared with our own modification of a plate diffusion technique using Sarcina lutea and yeast extract agar. Six of the eight methods were less sensitive than originally reported, and five of them were considered unsuitable for use in clinical microbiology practice. The remaining three methods together with the S. lutea/yeast extract modification were used to assay chloramphenicol in 20 samples of serum. Twenty samples of cerebrospinal fluid were also assayed by the S. lutea/yeast extract method. Our results indicate that only the Bacillus subtilis (sensitivity 6x0 mg/l) and the S. lutea (sensitivity 2x5 mg/l) diffusion methods are suitable for use with clinical samples in routine practice. The problems of chloramphenicol toxicity, appropriate dosage regimens, and the need for assay of the drugs are considered.

Bacillus subtilis↗
PubMed1976

Bacterial flora of the healthy skin in children.

The normal bacterial flora of the skin of 100 healthy children was studied in a paediatric practice with regard to the germ spectrum and the resistance constellation, by means of abrasion skin samples taken from the upper part of the back. Staphylococcus albus was by far the most predominant bacteria, followed by Sarcina bacteria and Corynebacteria. The high degree of pervasion of this physiological cutaneous flora with antibiotic-resistant strains increases the suspicion that in the case of florid bacterial skin infections in children, subsequent to traumatic or inflammatory changes of the integument, the antibiotic resistance of the normal resident flora can be transferred to the pathogenic flora. In view of the endemic resistance of pathogenic and saprobic strains of bacteria against practically all antibiotics at present used in paediatric practice, it is to be urgently recommended that instead of antibiotics topical braod-spectrum antimicrobial agents (antiseptics), e.g. clioquinol, triclosan, etc., should preferably be used.

Anti-Bacterial Agents↗
PubMed1996

In vitro antimicrobial activity of Pistacia lentiscus L. extracts: preliminary report.

The in vitro antimicrobial activity of Pistacia lentiscus L. extracts was determined. Pistacia lentiscus L. extracts were tested on bacteria (Sarcina lutea, Staphylococcus aureus and Escherichia coli) and fungi (Candida albicans, Candida parapsilosis, Torulopsis glabrata and Cryptococcus neoformans). Of the different plant extractions, decoctions showed the best antibacterial activity, but the activity against fungal cells appears to be much more interesting.

Anti-Bacterial Agents↗
PubMed1978

Anti-riboflavin activity of 8N-alkyl analogues of roseoflavin in some Gram-positive bacteria.

Three new 8N-alkyl analogues of roseoflavin (MM), i.e., 8-ethylamino- (EH), 8-methylethylamino- (ME), 8-diethylamino-8-demethyl-D-riboflavin (EE), their tetraacetates, and 8-amino-8-demethyl-D-riboflavin (HH) tetraacetate, were synthesized. A relation between the anti-riboflavin activity and the chemical structure of 8N-alkyl analogues (8N-methyl, ethyl) was studied by a restoration by riboflavin (RF) of inhibitory effect of the analogues on a growth of Gram-positive bacteria, i.e., Sarcina lutea, Bacillus cereus, and Staphylococcus aureus. The inhibitory effect of most of the analogues was restored by RF. But in some cases, i.e., 8-methylamino-8-demethyl-D-riboflavin (MH) in Sar. lutea and MM in Staph. aureus, the effect was not completely restored. Apparently, the inhibition in early phase of growth was restored, but the maximum growth was still suppressed. The non-alkylated amino analogue (HH) showed only unrestorable suppression of maximum growth in Sar. lutea. Of restorable effect by RF of N-alkyl analogues, approximate decreasing orders of anti-RF activity were as follows. Dialkylated analogue greater than monoalkylated. HH showed insignificant anti-RF activity. In each group, methylated analogue greater than ethylated. In B. cereus monoalkylated analogues, and in Staph. aureus monoalkylated and EE showed no significant inhibitory effect. Redox potentials of the N-alkyl analogues were measured, and a definite relation between the chemical structure and the potential was found (RF = EE greater than ME greater than MM greater than HH greater than EH greater than MH). But the anti-RF activity of the analogues was not completely explained by the difference of the redox potential from RF.

Anti-Bacterial Agents↗
PubMed1979

Anti-riboflavin activity of 8-O-alkyl derivatives of riboflavin in some Gram-positive bacteria.

Two new 8-O-alkyl derivatives of riboflavin (RF), i.e., 8-methoxy- (MOF), and 8-ethoxy-8-demethyl-D-riboflavin (EOF), their tetraacetate, and the tetraacetate of 8-hydroxy-8-demethyl-D-riboflavin (HOF) were synthesized. The anti-RF activity of MOF, EOF and HOF was estimated from the ratio CR/CI, where CI is the concentration of test flavin added to the culture medium and CR is the minimum concentration of RF needed to restore the growth inhibition. Their activity was also compared with that of roseoflavin (RoF). The decreasing order of anti-RF activity was as follows: MOF greater than RoF greater than EOF in Sarcina lutea: RoF greater than MOF greater than EOF in Bacillus cereus and Staphylococcus aureus. HOF showed no activity in any of the bacteria tested. The redox potential of these compounds decreases as follows: RF greater than RoF greater than EOF greater than MOF greater than HOF, and the RF activity of MOF and EOF could be explained by the redox potential difference between these compounds and RF.

Bacillus cereus↗
PubMed1970

The effect of high vacuum on oxidative reactions in bacteria and the activity of certain enzymes.

Among the physical factors which might influence micro-organisms one of the most potentially interesting is high vacuum. The effect of high vacuum is less studied as compared with other physical factors. It is impossible to achieve, under laboratory conditions, a vacuum of the order 10(-16) mm Hg which is probably characteristic of space. Earlier, the effect of high vacuum was studied on different bacteria, yeasts, molds and algae. It appeared that spores and fungal conidia were not killed by high vacuum. Later, the effect of high vacuum on physiological processes in micro-organisms was studied. The ability to oxidize glucose or ethanol was studied with Sarcina flava and Bacillus simplex cells after they were subjected for 72 hr to vacuum (10(-8) to 10(-9) mm Hg). The oxidation rate was followed polarographically. The oxidative ability of S. flava cells diminished [correction of dimished] after their subjection to vacuum, while B. simplex spores were unchanged in that respect. The following crystalline enzymes were subjected for 72 hr to the same vacuum: alpha-amylase, catalase, ribonuclease, trypsine and urease. Then the activity of the above enzymes was tested on corresponding substrates. Not a single enzyme was totally inactivated. About 50% of activity was lost with alpha-amylase; 25--35% of activity with catalase, ribonuclease and urease. Trypsine retained its total activity. Thus, high vacuum cannot be listed among factors rapidly inactivating enzymes of micro-organisms.

Bacillus↗
PubMed1976

Antibacterial and antitumor activity of some derivatives of ureidosuccinic acid.

The effect of several derivatives of ureidosuccinic acid on the growth of Escherichia coli 387, Staphylococcus aureus strain 209 and its mutant UV-2, Sarcina lutea, Candida tropicalis and Neurospora crossa 9863 as pre-screening systems for antitumor activity was studied. It was found that dihydrazide of D,L-ureidosuccinic acid (DHUA) had a marked antibacterial activity. The inhibitory effect of DHUA on N. crassa could not be removed by aspartic acid, ureidosuccinic acid, dihydroorotic acid, orotic acid, uracil or cytosine. DHUA suppressed the growth of Myeloma P-8 by 38%, that of Sarcoma 180 by 12% and that of Yoshida sarcoma by 19%. No effect was found on the growth of Lymphosarcoma Pliss.

Antineoplastic Agents↗
PubMedDate not supplied

[Quantitative study and typization of aerobic bacteria in the conjunctival sacs of healthy eyes].

We carried out a qualitative and quantitative bacteriological research into the conjunctives of 220 subjects in good health mostly women (180) exempt from recent or previous eyes diseases. In order to gather conjunctival bacteria we employed with a few variations Hadley's et al. (1973 [12]) method. Among the calculated bacterial flora we found as the most numerous ones Staphylococcus epidermidis (mean valor 1960/ml), then Streptococci (480/ml), Micrococci (465/ml), Diphteroides (360/ml), Enterococci (77/ml) and numerous (700/ml) Gram-positives Cocci like Sarcina, Micrococcus tetragenes, Diplococcus, and very rarely bacterial genera Gram-negatives or Iphomycetes, also numerically inferior as regards the researches of other Authors, because owen study is carried by into eyes exempt from diseases.

Adolescent↗
PubMed1991

Some pharmacokinetic data on miocamycin II. Concentrations in gynaecological tissues.

The gynaecological tissue levels of miocamycin were studied in ten female patients after pre-operative administration. The patients were divided into two groups on the basis of the scheduled sampling time. All the patients received 4200 mg of the drug, divided in 7 tablets of 600 mg each every 8 h, last administration 2 or 3 h before surgery. The tissue levels, obtained by means of the microbiological method (Sarcina lutea 9341), were frequently higher than those obtained in the serum at the same time. Miocamycin reached the highest concentration in the endometrium (2.5 micrograms/g) and the lowest in the vagina (1.1-0.8 micrograms/g).

Biological Assay↗
PubMed1975

Survey of antibiotic residues in Canadian slaughter animals.

Kidneys and urine of cattle, swine, sheep and chickens were tested for bacterial growth inhibitors using Bacillus subtilis and Sarcina lutea as test organisms. Results were as follows: 211 beef kidneys four positive, 611 swine kidneys five positive, 27 sheep and 120 chicken kidneys all negative, 2108 beef urine 76 positive, 2409 swine urine 186 positive, 176 sheep urine 17 positive. Strongest reactions were obtained with B. subtilis on phosphate buffered pH 6.0 plates. Larger zones were produced by urine from injected animals than by tissue samples.

Abattoirs↗
PubMed1988

[The distribution of microorganisms in household kitchens. I. Problems, experiments, results].

Epidemiological investigations have demonstrated that insufficient hygiene in households results in increasing health hazards. In order to be able to recommend ways to disrupt infection chains it was necessary to explore the most important pathways of cross-contamination. Housewives without special training were instructed to prepare a complete meal, in the kitchen of a modern, vacant apartment. Among the raw products provided was minced meat, contaminated with Sarcinae (unbeknown to the housewives) in a quantitative manner. When cooking and cleaning procedures were completed we analysed household utensils and surfaces by Rodac impressions and swabs. The test organisms could be detected on all inspected surfaces and on the dining-table with albeit different frequencies. The following locations showed an especially high degree of cross-contamination: a) working-surfaces, especially boards of wood and plastics. b) cutting-machines, c) kitchen-machines. These results agree with literature data. By careful disinfection, i.e. by application of a 0.5% solution of hypochlorite, the contaminations could be removed. We assessed this when arranging the kitchen for the next test. Since it is impossible to practise disinfection procedures in a household kitchen on the same scale as in an operating room, we tried to achieve at least a limited disinfection by household cleansers with germicidal properties. In our opinion a minimum reduction of five log stages, demanded in the medical area, can not be achieved in a household kitchen and indeed it is not necessary. A reduction of the microbial counts to 10% of the original value would already be useful, as toxic levels of microbial counts will be reached later especially when there is simultaneous refrigeration. Correct dosage proved to be one of the main practical problems because a discrepancy exists between the low concentration of tensids, necessary for cleaning, and the relatively high dose necessary for germ-killing compounds. Diluted DOMESTOS proved to be a cleaning agent and germicide, but was, however, blamed for chlorine odour, especially when diluted with warm water. A greater acceptance level was reached with a peroxide-containing cleanser, which, however, was not sufficiently germicidal, when applied in diluted form. The concentrated formulation was more effective in everyday experience, but for this housewives had to wear rubber gloves. This was reported to be complicated and uncomfortable, and indeed the search for better formulations must be continued. (In communication II comparisons with the bibliography and hygienic consequences will be published.)

Cooking and Eating Utensils↗
PubMed1988

[The distribution of microorganisms in household kitchens. II. Evaluation of results and hygienic inferences].

The very high morbidity rates of Enteritis infectiosa diseases demand improved prophylactic measures. An important indication of the source of these illnesses is the fact that infections in private households are about three times more frequent than in canteens. Indeed, the rise in morbidity is undoubtedly caused by inadequate treatment of raw products, of meal rests and by insufficient heating processes. Furthermore, in household kitchens no efforts are made to interrupt infection chains, and disinfections are considered as superfluous and housewives are content if their kitchens appear to be clean. The aim of our study performed in a normal household kitchen, was to investigate cross-contamination caused by pathogens, introduced into the kitchen from outdoors. A further aim was to establish the main sources of contamination in order to be able to recommend practical disinfection procedures. The main fields of contamination discovered when 55 meals prepared were: a) working surfaces (including boards of wood and plastics) b) kitchen- and cutting-machines. The amount of test organisms (Sarcinae), introduced into the kitchen (unbeknown to the housewives) by experimentally contaminated minced meat was only reduced by common cleaning procedures, in sofar as nearly half of the original contaminations could be demonstrated to be still present. However, when the normal cleanser was replaced by one containing hypochlorite, and with retention of the same working routines, about 90% bacteriologically clean surfaces were determined. In this way it could be demonstrated that infection chains can be interrupted. It is, however, not correct to compare the efficiency of these procedures with the efficiency of disinfection, according to the Federal Infectious Diseases Act (Bundesseuchengesetz). On practical application of these experiences it must be borne in mind that housewives should not be forced to apply medical disinfection procedures: indeed, traditional and practised cleaning methods should be retained, as far as possible. We recommend therefore that manufacturers supply household cleansers with an anti-bacterial additive, after its application in the kitchens working surfaces and machines are bacteriologically clean. Additionally housewives should be appropriately informed about the necessity of these manipulations. We consider minimization of toxicity and a thorough environmental compatibility of formulations to be self-evident.

Cooking and Eating Utensils↗
PubMed1979

Determination of virginiamycin in feeds.

Virginiamycin was extracted from the feed by ethanol-pH 2.5 phosphate buffer (1 + 1). The pH during extraction was adjusted (when necessary) to between 4 and 5. Sample dilutions and the standard dose response line were prepared to contain ethanol pH 6 phosphate buffer (2 + 8), and the test organism was Sarcina lutea. Three feeds (a poultry ration, a swine finishing ration, and a swine starter ration) showed virginiamycin recovery of 88.8--108.9% when standard solutions were added at concentrations of 4.54--90.8 g/ton. The coefficient of variation (4--20%) was larger for low potency feeds (10 g/ton) compared to the higher feeds (100 g/ton). Similarly, excellent recovery was obtained when the swine starter feed was fortified by a commercial premix. Amprolium, roxarsone, and monensin can be present at 20 times the concentration of virginiamycin with little or no interference in the antibiotic determination. Lasalocid at 10 times the concentration of virginiamycin caused a slightly positive bias (recovery, 107.4%).

Animal Feed↗
PubMed1977

Enumeration and identification of human leukemic lymphocytes by their natural binding of bacteria.

The recently described property of bacteria to bind to human lymphocytes was used to distinguish between normal and chronic leukemic lymphocyte (CLL) populations. Strains of the following bacteria were used in this study: Arizona hinshawii, Escherichia coli strains 1 and 2, Bacillus globigii, Brucella melitensis, Corynebacterium diphtheriae strains 1 and 2, Corynebacterium xerosis, Sarcina lutea, Staphylococcus aureus, and Staphylococcus epidermidis. For identification of immunoglobulin-bearing lymphocytes, a strain of E. coli that did not bind to human lymphocytes was coated with anti-human light-chain antibody. Labeling of lymphocytes with bacteria was promoted by centrifugation. In the eight CLL patients studied, in which greater than 90% of the lymphocytes were leukemic cells, 52 to 77% were labeled by anti-human light-chain antibody-E. coli, 80 to 93% were labeled by Br. melitensis, and 78 to 95% were labeled by E. coli 1 compared to 11 to 24, 11 to 22, and 30 to 44%, respectively, in normal individuals, Thus, Br. melitensis, E. coli 1, and the anti-human light-chain antibody-E. coli may have diagnostic value for CLL. The percentage of the lymphocyte population that bound each of the other bacteria varied from patient to patient. Preliminary results obtained by studying the pattern of binding of E. coli 2, B. globigii, Sa. lutea, or S. aureus by leukemic lymphocytes suggest that categories of CLL patients may be distinguished by this method.

Antibodies, Anti-Idiotypic↗
PubMed1976

Radioimmunoassay of neocarzinostatin, an antitumor protein.

Antibodies directed toward the antitumor protein neocarzinostatin (NCS) have been produced in a rabbit by immunization with a highly purified NCS preparation. The antiserum was monospecific and reversed the antibacterial activity of NCS against Sarcina lutea. It cross-reacted with chemically modified derivatives of NCS and mitomalcin but failed to cross-react with macromomycin. A radioimmunoassay procedure has been developed utilizing the antiserum and a biologically active 125I-labeled derivative of NCS. The lower limit of detection by this radioimmunoassay, which involves a double antibody technique for the separation of antibody-bound and free antigen, was 1 X 10(-13) mole. The sensitivity of the assay is such that serum levels of NCS can be determined accurately after administration of the drug to rats at a single dose of 2 mg/kg. Since NCS is now undergoing clinical trial, the radioimmunoassay of the drug will be a valuable tool in clinical pharmacological studies.

Animals↗