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Bactericidal effect of 15-deoxyspergualin on Staphylococcus aureus.

15-Deoxyspergualin (DSG), an immunosuppressive agent used in organ transplantation, exerts metabolic and antiproliferative effects on methicillin- and gentamicin-resistant Staphylococcus aureus and other bacteria (e.g. Sarcina lutea, Bacillus subtilis, Shigella sonnei, Salmonella typhi and Citrobacter freundii). DSG, at the concentration of 20 mg/l, depleted intracellular putrescine and spermidine in S. aureus to 43 and 40% of the controls, respectively. In these polyamine-depleted S. aureus cells, the synthesis of protein, DNA and RNA was decreased to 20, 85, 78% of the controls. The minimal inhibitory concentrations (MIC) of DSG for growth of S. aureus, S. lutea, B. subtilis, S. sonnei, S. typhi and C. freundii were 17, 13, 7, 15, 4, and 29 mg/l, respectively.

Bacteria↗

Characterization of lactic acid bacteria and other gut bacteria in pigs by a macroarraying method.

Lactic acid bacteria (LAB) consist of many genera, Gram-positive, and nonspore-forming micro-organisms; some members being used as probiotics while some others have negative effects on pig health. Bacterial species in the gastrointestinal tract can produce antibacterial substances, reduce serum cholesterol in their host, or can be responsible for growth reduction, diarrhea, and intestinal epithelial damage. It is therefore important for the pig industry to evaluate the impact of food and farm management on the presence of "good" or "bad" bacteria and the risk for consumers. This articles focuses on the molecular identification of gut microflora species following different diets given to pigs in UK and correlating the data on growth, health, and welfare. First of all, pig feces were individually collected from sows before and after farrowing and also from piglets before and after weaning over several months. Bacteria colonies were grown on MRS agar plates from feces and DNA was extracted (QIAamp DNA stool kit) and amplified using 16S rDNA (27f and 519r) primers. DNA sequencing and sequence alignment allowed us to identify species-specific zones, which were used as probes in a macroarray system also known as reverse line blot hybridization. Some probes were found to be species specific for the following species: Lactobacillus acidophilus, L. animalis, L. gallinarum, L. kitasanotis, L salivarius, Streptococcus alactolyticus, S. hyointestinalis, and Sarcina ventriculi. Actual studies are now focusing on the impact of diets of the microflora in different gut parts and at different stages of the animal's life.

Animals↗

Prospective evaluation of the incidence of bacteremia after protected specimen brushing in ICU patients with and without pneumonia.

To test the hypothesis that the use of protected specimen brushing (PSB) via flexible bronchoscopy does not predispose to bacteremia in ICU patients, we prospectively performed aerobic and anaerobic blood cultures immediately following bronchoscopy with PSB. A total of 123 episodes in 68 consecutive patients with suspected pneumonia were analyzed. Blood cultures were negative in 110 cases (89 percent) and positive in 13 cases (11 percent) (p < 0.001). Twelve of these 13 patients with positive blood cultures had quantitative PSB specimen cultures showing nonsignificant growth (< 10(3) CFU/ml). In nine patients, the bacteria recovered from blood cultures (coagulase-negative staphylococci or sarcina) were considered nonpathogenic according to conventional criteria. Blood cultures grew a Staphylococcus aureus in two patients with previously documented staphylococcal septicemia. In one patient with no identifiable site of infection, the blood culture yielded Enterococcus faecalis. The only patient with both a positive blood culture and PSB culture results indicating pneumonia had different organisms recovered from the two samples. Blood cultures taken after PSB in the 17 other episodes of pneumonia (PSB specimen cultures > or = 10(3) CFU/ml) were negative. At the time of brushing and blood sampling for culture, none of these patients was receiving antibiotics active on the organisms found. In conclusion, the incidence of bacteremia after PSB in ICU patients seems very low even in patients with documented pneumonia. Substantial savings would result from not performing routine blood cultures after PSB.

Aged↗

Production of a macromomycin (MCR)-monoclonal antibody conjugate and its biological activity.

Macromomycin (MCR), an unique membrane-reactive anticancer antibiotic, was incubated with murine monoclonal anti-HLA IgG1 antibody (H-1) in the presence of carbodiimide. The resulting mixture was fractionated with a Sephadex G-200 column. The first and second fractions were shown to contain MCR-(H-1) conjugate by the elution profile, as well as by the Sarcina lutea growth inhibition assay and Ouchterlony double-diffusion method. A membrane immunofluorescence test with anti-MCR and anti-mouse IgG antibodies demonstrated specific localization of MCR-(H-1) on the surface of HLA-bearing NALL -1 cells. MCR-(H-1) inhibited the growth of HLA-lacking NS-1 cells statistically less effectively than MCR alone (p less than 0.01). On the other hand, the conjugate and free MCR equally inhibited the growth and 3H-TdR incorporation of HLA-bearing NALL -1 cells. These results indicate that the antibody-bound MCR retained both MCR and antibody activities, and thus exerted antibody-targeting MCR cytotoxicity in vitro.

Anti-Bacterial Agents↗

Kinetic profile of miocamycin in middle ear fluid and mucosa.

A group of 18 patients, before undergoing otoplastic surgery, was treated with miocamycin 600 mg in a single dose; middle ear tissue samples were collected at 2, 3, 4 and 6 h. A group of 20 patients suffering from secretory otitis media (SOM) was also treated with 600 mg of miocamycin in a single dose; collections of middle ear fluid (MEF) were performed at 1, 2, 4 and 6 h. Blood withdrawals from all patients occurred at the same time intervals. The miocamycin determination was performed by means of a microbiological method employing Sarcina lutea ATCC 9341. The highest serum levels (1.06 +/- 0.13 mg/l) were found at 2 h in the group from which middle ear mucosa had been withdrawn and at 1 h (2.2 +/- 0.5 mg/l) in the group from which the MEF had been collected; at 6 h miocamycin could be determined in both groups, with values ranging from 0.1 to 0.2 mg/l. In middle ear mucosa the miocamycin concentration was 1.52 +/- 0.27 mg/kg at 2 h and 0.2 +/- 0.09 mg/kg at 6 h. In MEF, the miocamycin concentration was 2.1 +/- 0.27 mg/l at 1 h and 0.24 +/- 0.05 mg/l at 6 h. The miocamycin concentration determined at 1 h in MEF was virtually equal to that in serum at the same time. At the following experimental times of withdrawal, the miocamycin concentration in mucosa specimens, as well as in MEF samples, proved to be markedly higher than values simultaneously found in serum.

Adult↗

Persistence of antibiotics in milk from cows treated late in the dry period.

Nineteen cows were treated via intramammary infusion at various times prior to calving with one of two dry cow antibiotic preparations (cloxacillin or penicillin). Milk samples from these cows after calving were analyzed for antibiotic residues by the Sarcina lutea cylinder plate method. Eight cows were treated with cloxacillin was detected in the milk of only two cows after calving and then for only one milking, so no problem should exist with cloxacillin if the manufacturer's recommendations are followed. Penicillin residues were not detected in milk samples taken from cows treated more than 9 days prior to calving with the dry cow penicillin preparation. However, seven of nine cows treated with penicillin up to 9 days prior to calving had detectable penicillin residues in the milk from treated quarters after calving. Only one cow had detectable penicillin residues longer than the manufacturer's recommended milk withholding time of 96 h, and this cow was treated only 3 days prior to parturition. Normal routine on a dairy farm and adherence to manufacturer's recommendations should result in no antibiotic residue problems in milk from cows treated during the dry period for mastitis control with either cloxacillin or penicillin.

Animals↗

Mechanism of DNA degradation induced by neocarzinostatin in Bacillus subtilis.

When logarithmically growing Bacillus subtilis cells were exposed to the antitumor protein, neocarzinostatin (NCS), at a concentration of 50 mug/ml, cellular DNA was gradually degraded into an acid-soluble form (up to 60 percent of total DNA). The degradation appeared to initiate at the growing regions of DNA and to proceed sequentially from the nascent regions to preexistent DNA. Concomitantly with, or perhaps as a consequence of, the degradation of growing regions, DNA detached from the cell membrane and started to show single-strand nicks within 30 minutes after exposure of the cells to NCS, whereas double strand scission in the DNA became detectable in about 90 minutes. Such endonucleolytic breaks in DNA eventually gave rise to the formation of double-stranded DNA fragments of a single-size class (30-S) as determined by sedimentation in either neutral or alkaline sucrose gradients. In contrast to previous results with Sarcina lutea, the NCS-induced DNA degradation was stimulated by chloramphenicol in B. subtilis and the DNA fragment were not the final breakdown products, but were further degraded into acid-soluble materials.

Antibiotics, Antineoplastic↗

[Examination of market carps for residues of antimicrobial agents].

The residues of inhibitory substances in the muscle, hepatopancreas, bile, and pyloric caeca of 30 carps from ponds south of Brno were examined by the microbiological diffusion method. The following testing micro-organisms were used: B. cereus var. mycoides (ATCC 11778), B. subtilis (ATCC 6633), Sarcina lutea (ATCC 9341), and Micrococcus flavus (ATCC 10240). Residues of inhibitory substances were not found in any of the muscle samples tested. The rest of the tested tissues showed, in some cases, small inhibition zones which seem to be due to the presence of natural antimicrobially active substances in the bile and gastro-intestinal tract of the fish rather than to the persistence of the residues of drugs given to the fish during their life.

Animals↗

The effect of storage at 4 degrees C on antibiotic residues in kidney and meat tissues of dairy cows.

A method is described for the quantitative assay of antibiotic residues in the body of slaughtered farm animals by means of 3 types of agar media and 3 test organisms. With the help of programmed calculation procedures, data from large-scale tests for antibiotic residues could be analyzed accurately and rapidly. The concentrations of penicillin G, ampicillin, amoxycillin, cloxacillin, cephapirin, cephacetrile, neomycin, kanamycin, and oxytetracycline in the kidney, and also of tylosin in meat from the diaphragm muscle of cattle treated parenterally with these antibiotics were measured periodically in samples kept at 4 degrees C for up to 7 days after slaughter. The concentrations of penicillin G, ampicillin, amoxycillin and the cephalosporins in the kidney decreased rapidly upon storage whereas the levels of the other antibiotics remained essentially unchanged. Antibiotic stability in the meat was considerably greater than in the kidney upon storage for 4 days, and neomycin meat tissue levels were not reduced during storage for up to 144 hours. Results are discussed in relation to the conduct of the official qualitative Sarcina lutea Kidney Test and the most desirable procedure for preparing meat samples for assay.

Animals↗

[Distribution of erythromycin following intramuscular administration in laboratory animals].

Levels of erythromycin in the blood and organs of rabbits, rats and mice were determined after the i. m. implantation of the drug Erythromycin inj. ad usum vet. at doses of 10 mg kg-1 live weight (rabbits), and/or 25 mg kg-1 (rats) and 1 mg pro toto--approximately 40 mg kg-1 (mice). A diffusion plate method and the germ Sarcina lutea CCM 552 were used to find out the antibiotic in samples. Sensitivity of the method used was 0.025 microgram ml-1 (g). In rabbits the level of the antibiotic was maintained in the blood and examined organs and tissues (lungs, liver, kidney, muscles and the wall of the small intestine) by the above dose for more than 12 hours--except liver. The values obtained for the organs and tissues were mostly higher than those obtained for the blood. The highest levels were found in the kidneys and lungs. Neither did the dose of 20 mg kg-1 l. w. maintain the significant level of erythromycin in the blood serum in the course of 24 hours. The results obtained in rats and mice almost coincided with the distribution of erythromycin in the rabbit organisms, and the affinity of this antibiotic with the pulmonary tissue was proved.

Animals↗

[Passage into normal salivary, lacrimal and nasal secretions of ampicillin and erythromycin administered intramuscularly].

The authors have measured, with the agar diffusion technique, the antibiotic concentrations in the serum as well as in the normal nasal, lachrymal and salivary secretions, after an intramuscular injection of 500 mg of ampicillin and of 200 mg of erythromycin. Bacillus subtilis is used as test germ for dosage of ampicillin and Sarcina lutea for the dosage of erythromycin. An important quantity of ampicillin (0.5 to 2.5 microgram/ml) passes into the nasal secretion. Its passage into the saliva is very irregular and is usually very slight. Bacillus subtilis is not suitable for systematically studying the ampicillin concentration in the lachrymal secretion, which displays a strong, inherent inhibitory effect on this micro-organism. Important concentrations of erythromycin are found in the three secretions: 0.5 to 3.25 microgram/ml in the nasal mucus and in the lachrymal secretion, 0.3 to 4.5 microgram/ml in saliva.

Ampicillin↗

Pharmacokinetic interpretation of erythromycin and tylosin activity in serum after intravenous administration of a single dose to cows.

The distribution and elimination kinetics of erythromycin and tylosin, which are macrolide antibiotics, were studied in healthy cows. A single dose (12-5 mg/kg) of drug was administered as an intravenous bolus, and blood samples were collected at precisely timed intervals. The standard cylinder plate bioassay method using Sarcina lutea as test organism was employed to determine antibiotic activity in the serum. The results suggested that these drugs are distributed in at least two kinetically distinct body compartments. By use of established mathematical techniques, values were assigned to the individual rate constants controlling distribution between the central and peripheral compartments and to the rate constant controlling overall elimination (beta) of each drug from the body. The calculated overall tissue to serum drug level ratios (k12/k21) after apparent distribution equilibrium was attained were 2-28 and 2-05 for erythromycin and tylosin, respectively. The half-life (mean+/-SD) of erythromycin was 3-16 h+/-0-44, while that of tylosin was 1-62 h+/-0-17. The total body clearance (ml/kg/min) values were 2-88+/-0-47 for erythromycin and 7-8+/-2-95 for tylosin. Analogue computer simulated curves of the antibiotic levels in the central and tissue compartments as wel as an elimination curve were generated. The tissue level of erythromycin reached a peak of 43 per cent of the dose at 67 min. At 6 h, the percentages of the dose of erythromycin in the central and tissue compartments and eliminated were 6, 19 and 75, respectively. The peak level of tylosin in the tissue compartment (26-5 per cent of the dose) was present at 30 min. At 4 h, 1 and 5 per cent of the dose were contained in the central and peripheral compartments, respectively, while 94 per cent had been eliminated. This single dose study provides information which is essential for the design of a satisfactory dosage regimen.

Animals↗

Diagnostic evaluation of gastric analysis (test meal) in Hungary.

On the basis of information received from 128 medical wards, the frequency of gastric analysis (test meal) used as a clinical diagnostic procedure, the spreading of the different methods and other problems connected with the examination are analysed. In about 3/4 of the institutions the test meal is performed in patients with all kinds of gastric complaint. This incidence decreases to 50% if the X-ray finding is positive. As acid-stimulants caffeine and a moderate dose of histamine are used most frequently. Of the data supplied by the test meal, the value for acid concentration (acidity) is registered by most of the examiners. The authors consider it as suprising that outdated methods such as a qualitative lactic acid assay (47%), isolation of Boas-Oppler's bacillus (25%) and of sarcinas (21%) are still in frequent use. The data are compared to the results of a similar survey made in England.

Gastric Acidity Determination↗

Female genital tissue concentrations of roxithromycin.

A new macrolide, roxithromycin, appears to have some interesting pharmacokinetic characteristics dissimilar to those of erythromycin, e.g. it has the unusual property of a long serum half-life and the possibility of once daily dosing. Moreover tissue levels are higher than those obtained with other macrolides, and the drug is still present in the tissues up to 24 h. after dosing. The pharmacokinetics of roxithromycin were studied in 36 women undergoing gynecological surgery, and divided by means of the time of sampling into six different groups. Each group received an initial 300 mg dose followed by 8 successive doses of 150 mg at 12 h intervals. Blood and tissues samples were taken during surgery at 2, 6, 9, 12, 18 and 24 h after the last dose of roxithromycin and the patients were then allocated to one of the six groups by means of the sampling time. Tissue fragments were obtained from ovary, fallopian tubes, endometrium, myometrium, cervix and vagina immediately after the surgical resection of the organs. Tissue and serum concentrations were determined by the microbiological method using Sarcina lutea ATCC 9341; the lowest limit of detection was 0.01 microg/ml or 0.01 microg/g. Roxithromycin reached the highest concentration at the 9th hour after last administration and its tissue levels would encourage a wide use of this drug as a satisfactory alternative to tetracyclines for the therapy of some gynecological infections.

Administration, Oral↗

The international standard for aureomycin.

In 1950, the Department of Biological Standards, National Institute for Medical Research, London, was authorized by the WHO Expert Committee on Biological Standardization to proceed with the establishment of an International Standard for Aureomycin. A 100-g batch of aureomycin was obtained and was compared with the Standard Preparation of Aureomycin of the United States Food and Drug Administration (FDA) in a collaborative assay in which six laboratories in five countries participated.In all, 30 assays were carried out; 26 of these were done by biological methods, using Sarcina lutea, Bacillus pumilus, Staphylococcus aureus, or Bacillus cereus, and the remaining four by physicochemical methods. The results were subjected to standard methods of analysis, and the overall weighted mean potency (calculated from the biological assays only) was 1.0139, with limits of error of 99.5% to 100.5%.Since the International Standard is 1.39% more potent than the FDA Standard Preparation, it is probable that the latter contains a small amount of inert material; it is also possible that the International Standard itself is not 100% pure. For most practical purposes, however, both preparations may be regarded as substantially pure, and it is considered that to alter the present practice of quoting aureomycin dosage in metric units of weight would be inadvisable. Nevertheless, since the International Standard may not be a pure substance, a unit notation-for use where required in bioassays-is desirable, and the International Unit of Aureomycin has therefore been defined as the activity contained in one microgram of the International Standard.

Biological Assay↗

Synthesis and biological activity of new heterocyclic structures: [1,3]thiazino[2,3-i]purine, thiazolo[3,2-c] [1,2,3]triazolo[4,5-e]pyrimidine and [1,2,3]triazolo [4', 5': 4,5] pyrimido[6,1-b][1,3]thiazine.

Some derivatives of thiazolo[3,2-c]pyrimidine, pyrimido[6,1-b][1,3]thiazine, thiazolo[2,3-i]purine, [1,3]thiazino[2,3-i]purine, thiazolo[3,2-c][1,2,3]triazolo[4,5-e]pyrimidine and [1,2,3]triazolo[4',5':4,5]pyrimido[6,1-b][1,3]thiazine were prepared. The compounds were tested for antimicrobial and antimycotic activity on a number of strains, namely, Escherichia coli, Proteus vulgaris, P. mirabilis, Pseudomonas aeruginosa, Salmonella sp., Staphylococcus aureus, Streptococcus faecalis, Bacillus subtilis, Sarcina lutea, Candida albicans, Aspergillus sp., and for antiviral activity on Herpes simplex virus, Type 1 (HSV-1), Vesicular stomatitis virus (VSV) and Coxsackievirus B5 (CoxB5). The compounds proved to be devoid of activity against viruses, mycetes and gram-negative bacteria, while some of them exhibited a modest activity against gram-positive bacterial strains.

Animals↗

Influence of nutrients on biomass evolution in an upflow anaerobic sludge blanket reactor degrading sulfate-laden organics.

This paper describes the effect of the nutrients iron (Fe), nickel (Ni), zinc (Zn), cobalt (Co), and molybdenum (Mo) on biomass evolution in an upflow anaerobic sludge blanket (UASB) reactor metabolizing synthetic sulfate-laden organics at varying operating conditions during a period of 540 days. A bench-scale model of a UASB reactor was operated at a temperature of 35 degrees C for a chemical oxygen demand-to-sulfate (COD/SO4(2-)) ratio of 8.59 to 2.0, a sulfate loading rate of 0.54 to 1.88 kg SO4(2-)/m3 x d, and an organic loading rate of 1.9 to 5.75 kg COD/m3 x d. Biomass was characterized in terms of total methanogenic activity, acetate-utilizing methanogenic activity, total sulfidogenic activity, acetate-utilizing sulfidogenic activity, and scanning electron microscopy (SEM). Nickel and cobalt limitation appears to affect the activity of hydrogen-utilizing methane-producing bacteria (HMPB) significantly without having an appreciable effect on the activity of acetate-utilizing methane-producing bacteria (AMPB). Nickel and cobalt supplementation resulted in increased availability and, consequently, restoration of biomass activity and process performance. Iron limitation and sulfidogenic conditions resulted in the growth of low-density, hollow, fragile granules that washed out, causing process instability and performance deterioration. Iron and cobalt supplementation indicated significant stimulation of AMPB with slight inhibition of HMPB. Examination of biomass through SEM indicated a population shift with dominance of sarcina-type organisms and the formation of hollow granules. Granule disintegration was observed toward the end of the study.

Bacteria, Anaerobic↗

Synthesis and biological activity of pyrimido[2,1-b][1,3]thiazine, [1,3]thiazino[3,2-a]purine and [1,2,3]triazolo[4,5-d][1,3]thiazino[3,2-a]pyrimidine derivatives and thiazole analogues.

Some series of thiazolo[3,2-a]pyrimidine, pyrimido[2,1-b] [1,3]thiazine, thiazolo[3,2-a]purine, [1,3]thiazino[3,2-a]purine, thiazolo[3,2-a][1,2,3]triazolo[4,5-d]pyrimidine and [1,2,3]triazolo[4,5-d][1,3]thiazino[3,2-a]pyrimidine derivatives, variously functionalized, were prepared. The compounds were tested for antimicrobial and antimycotic activity on a number of strains, namely: E. coli, Proteus vulgaris, P. mirabilis, Pseudomonas aeruginosa, Salmonella sp., Staphylococcus aureus, S. faecalis, Bacillus subtilis, Sarcina lutea, Candida albicans, C. tropicalis, Aspergillus sp., and for antiviral activity on Herpes simplex virus Type 1, Vesicular stomatitis virus and Coxsackievirus B5. The compounds proved to be devoid of activity against viruses and gram-negative bacteria, while some of them exhibited modest activity against gram-positive bacterial strains.

Anti-Bacterial Agents↗