Effect of protoplasting on antibiotic activity and resistance in the strain Streptomyces hygroscopicus 155]

The influence of protoplasting and protoplast regeneration in the presence of polyethylene glycol on antibiotic activity, components of antibiotic complexes and antibiotic resistance in Streptomyces hygroscopicus 155 was studied. It was shown that the protoplasting and protoplast regeneration influenced the antibiotic activity. The protoplast fusion resulted in increased isolation of variants with higher antibiotic activity. The processes also affected the components of the antibiotic complexes but had no effect on the strain resistance to some antibiotics.     

Hypotension caused by intravenous infusion of rifampicin and its relation to the administration schedule

It was shown in studies on animals that bolus administration of rifampicin induced hypotension whose severity depended on the rate of the antibiotic administration. When the antibiotic was administered in the 5-, 10- or 15-minute regimen in a dose of 10 mg/kg the maximum decrease in blood pressure was 44, 34 or 21% of the initial level and the maximum antibiotic concentration attained in the blood was 34.4, 27.2 or 22.6 micrograms/ml, respectively. With the infusion for 30 minutes, the maximum antibiotic concentration in the blood was 17.6 micrograms/ml and the blood pressure did not undergo any significant changes. When the rate of the antibiotic infusion was high there was pharmacokinetic heterogeneity of the blood serum and biophase which could lead to unpredictable results. After repeated administrations of rifampicin to the same animals pronounced tachyphylaxis to the antibiotic was noted, which manifested itself in decreasing of hypotension, though the serum antibiotic level was 1.5 to 2 times higher that the initial one. It was concluded that administration of rifampicin in the therapeutic dose equal to 10 mg/kg for 30 minutes was the most sparing regimen for the antibiotic bolus intravenous infusion. Gradual increase in the antibiotic dose and administration rate in patients is possible under careful control of blood pressure and pharmacokinetic studies.     

Antibiotics in surgical treatment of acute abscesses.

A four-way, double-blind, prospective trial of treatment of abscesses by incision, curettage, and primary closure with and without antibiotic cover (clindamycin injection before operation or capsules after operation, or both) was conducted. There was no appreciable difference in mean healing time between the patients given both the antibiotic injection and the antibiotic capsules and those given the injection and placebo capsules, whereas healing times in those given the placebo injection and antibiotic capsules or placebo only were appreciably longer. Four of the patients who were not given the antibiotic injection developed bacteraemia; one patient who was given the antibiotic injection also developed a bacteraemia, but this was caused by clindamycin-resistant bacteria. These results show that a single injection of an effective antibiotic before operation is sufficient to protect the patient against bacteraemia and permit optimum healing.     

Bacereutin, an antifungal antibiotic isolated from metabolites of Bacillus cereus CHU 130

An antifungal metabolite, bacereutin, was isolated from culture filtrate of Bacillus cereus CHU 130. The bacterium was isolated from soils collected in Changhwa County, Taiwan, and was grown in soybean meal-mannitol broth for production of the antibiotic metabolite. The antibiotic metabolite was isolated by adsorption column chromatography of Amberite XAD-2 and was purified by passing through the chromatographic columns packed with Dowex 50W-X8, Sephadex LH 20 and Biogel P-2. The antibiotic metabolite was soluble in water and 87% acetone, and was slightly soluble in methanol, but was not dissolved in n-propanol, n-butanol, acetone, benzene and ethyl acetate. The antibiotic metabolite was a heat-stable and ninhydrin-positive substance. The antibiotic activities of bacereutin were tested by means of the agar-diffusion plate method. The antibiotic metabolite inhibited the growth of Saccharomyces cerevisiae CHU 1, Paecilomyces variotii CHU 6, Rhizomucor miehei CHU 40 and Fusarium oxysportum CHU 98. Bacereutin was a ninhydrin-positive antifungal antibiotic.     

耐药淋球菌相关核苷酸序列的克隆与初步分析*

为克隆淋球菌染色体耐药相关核苷酸序列,我们利用抑制性消减杂交技术构建耐药性淋球菌与标准参考菌株差异DNA消减库,从中筛选淋球菌耐药相关核苷酸序列。通过初步筛选,对克隆得到的DNA片段进行测序,经GenBank和淋球菌基因组序列库检索分析,发现5个未知新核苷酸序列。这些核苷酸序列可能与淋球菌染色体耐药性相关,将为研究淋球菌耐药性提供新的实验对象。     

Continuous antibiotic production by an immobilized cyanobacterium in a seaweed-type bioreactor

The filamentous cyanobacterium,Scytonema sp. TISTR 8208, which produces a cyclic peptide antibiotic, was cultivated for 20 d in a seaweed-type bioreactor containing anchored polyurethan foam strips. Cells immobilized onto the foam strips produced the antibiotic for only several days, and the secreted antibiotic disappeared very rapidly from the medium. Cells accumulated the antibiotic intracellularly in a growth-related manner, and secreted it in the stationary phase. Since the antibiotic has a stable physico-chemical nature, the cells seem to take it up and metabolize it. When continuous cultivation was attempted, stable production of the antibiotic was maintained in the bioreactor for 16 d at a dilution rate of 0.01 h^–1. Three times more antibiotic was produced in the continuous culture than in the batch culture by the 16th day.     

Influence of the culture medium on the production of iturin A by Bacillus subtilis

The production of iturin A by Bacillus subtilis was studied with respect to the composition of the culture medium. Increasing phosphate concentrations did not modify the antibiotic yield. Fructose, sucrose and mannitol were better carbon sources than glucose for antibiotic production. The nature of the nitrogen source was an important factor in the production of antibiotic. Among the amino acids which are components of iturin A, L-asparagine was the best substrate for the biosynthesis of iturin A; L-glutamine and L-serine were rather poor substrates while L-proline and D-tyrosine gave no antibiotic. Ammonium salts permitted good synthesis of antibiotic but the addition of calcium ions to the culture medium inhibited the excretion of antibiotic from the cells.     

Structure of pentaene antibiotic lavendofuseomycin

By its UV spectrum lavendofuseomycin, a macrolide pentaene antibiotic, was referred to the subgroup of adeopentaenes with the spectral symmetrical patterns. The antibiotic contains a carbonyl, the end and 4 isolated double bonds and hemiketal ring. The molecule is lacking sugar. After the hydroantibiotic oxidation 2-methylhexadecane dicarboxylic and 4'-methyloctanoic acids were isolated. The antibiotic carbon skeleton was asserted on the basis of the mass spectral analysis of the products of the antibiotic complete reduction and the products of the antibiotic retroaldol cleavage. Determination of the position of the isolated double bonds, localization of chromophore, oxygen functions and the position of the amino group in the molecule resulted from investigation of the antibiotic azonolysis products.     

Sampling and Pooling Methods for Capturing Herd Level Antibiotic Resistance in Swine Feces using qPCR and CFU Approaches

The aim of this article was to define the sampling level and method combination that captures antibiotic resistance at pig herd level utilizing qPCR antibiotic resistance gene quantification and culture-based quantification of antibiotic resistant coliform indicator bacteria. Fourteen qPCR assays for commonly detected antibiotic resistance genes were developed, and used to quantify antibiotic resistance genes in total DNA from swine fecal samples that were obtained using different sampling and pooling methods. In parallel, the number of antibiotic resistant coliform indicator bacteria was determined in the same swine fecal samples. The results showed that the qPCR assays were capable of detecting differences in antibiotic resistance levels in individual animals that the coliform bacteria colony forming units (CFU) could not. Also, the qPCR assays more accurately quantified antibiotic resistance genes when comparing individual sampling and pooling methods. qPCR on pooled samples was found to be a good representative for the general resistance level in a pig herd compared to the coliform CFU counts. It had significantly reduced relative standard deviations compared to coliform CFU counts in the same samples, and therefore differences in antibiotic resistance levels between samples were more readily detected. To our knowledge, this is the first study to describe sampling and pooling methods for qPCR quantification of antibiotic resistance genes in total DNA extracted from swine feces.     

A non-polyene antifungal antibiotic fromStreptomyces albidoflavus PU 23

In all 312 actinomycete strains were isolated from water and soil samples from different regions. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising strain,Streptomyces albidoflavus PU 23 with strong antifungal activity against pathogenic fungi was selected for further studies. Antibiotic was extracted and purified from the isolate.Aspergillus spp. was most sensitive to the antibiotic followed by other molds and yeasts. The antibiotic was stable at different temperatures and pH tested and there was no significant loss of the antifungal activity after treatment with various detergents and enzymes. Synergistic effect was observed when the antibiotic was used in combination with hamycin. The antibiotic was fairly stable for a period of 12 months at 4°C. The mode of action of the antibiotic seems to be by binding to the ergosterol present in the fungal cell membrane resulting in the leakage of intracellular material and eventually death of the cell. The structure of the antibiotic was determined by elemental analysis and by ultraviolet (UV), Fourier transform infrared (FTIR), nuclear magnetic resonance (NMR) and liquid chromatography mass spectra (LCMS). The antibiotic was found to be a straight chain polyhydroxy, polyether, non-proteinic compound with a single double bond, indicating a nonpolyene antifungal antibiotic     

Effect of support materials on antibiotic MSW2000 production by immobilized Streptomyces violatus

The production of an antibiotic by free and immobilized cells of Streptomyces violatus through entrapment or adsorption on different materials was investigated. S. violatus entrapped in Ca-alginate beads gave low antibiotic activity compared to the free cell or adsorbed cell, while the adsorption of S. violatus on sponge cubes yielded the highest antibiotic concentration after 4 days of incubation in static cultures. A starch concentration of 10 g/L was optimum for the production of the antibiotic by adsorbed cells. The weight and size of the sponge cubes used for immobilization influenced production of the antibiotic and the optimum weight and size of the sponge were 0.8 g and 1.0 cm(3), respectively, yielding a maximum antibiotic production of 280 mg/ml. Maximum antibiotic production was obtained at an initial pH value of 7.5 and in an inoculum size of 3 ml (spore suspension) per 50 ml. The production of the antibiotic in a fixed-bed bioreactor reached a maximum value after 2 days of incubation at a circulation rate of 30 ml/h. The immobilized cells in the bioreactor were reused seven successive times over a period of 14 days.     

Biological activity of an antibiotic produced by Myxococcus coralloides.

A strain of Myxococcus coralloides was isolated which produced an antibiotic active against Gram-positive bacteria and also against Neisseria sp at high levels of antibiotic. The antibiotic was bactericidal on sensitive growing bacteria. It was inactive on non-growing cells and also on cells whose growth has been stopped by addition of chloramphenicol. Strains of S. aureus resistant to several antibiotics, including penicillin and ampicillin, were also sensitive to this antibiotic.     

Production of the Antibiotic Phenazine-1-Carboxylic Acid by Fluorescent Pseudomonas Species in the Rhizosphere of Wheat

Pseudomonas fluorescens 2-79 and P. aureofaciens 30-84 produce the antibiotic phenazine-1-carboxylic acid and suppress take-all, an important root disease of wheat caused by Gaeumannomyces graminis var. tritici. To determine whether the antibiotic is produced in situ, wheat seeds were treated with strain 2-79 or 30-84 or with phenazine-nonproducing mutants or were left untreated and then were sown in natural or steamed soil in the field or growth chamber. The antibiotic was isolated only from roots of wheat colonized by strain 2-79 or 30-84 in both growth chamber and field studies. No antibiotic was recovered from the roots of seedlings grown from seeds treated with phenazine-nonproducing mutants or left untreated. In natural soils, comparable amounts of antibiotic (27 to 43 ng/g of root with adhering soil) were recovered from roots colonized by strain 2-79 whether or not the pathogen was present. Roots of plants grown in steamed soil yielded larger bacterial populations and more antibiotic than roots from natural soils. In steamed and natural soils, roots from which the antibiotic was recovered had significantly less disease than roots with no antibiotic, indicating that suppression of take-all is related directly to the presence of the antibiotic in the rhizosphere.     

Initiation of antibiotic production by the stringent response of Bacillus subtilis Marburg

Bacillus subtilis Marburg was found to produce an appreciable amount of an antibiotic in a synthetic medium. Antibiotic activity was produced in parallel with cell growth, and production stopped at the end of exponential growth. When the synthetic medium was supplemented with a small amount of Casamino acids, however, antibiotic was made only at the end of growth and in lesser amounts. The ability of cells to produce the antibiotic increased when stringent (rel+ = wild-type) cells underwent a partial stringent response. These conditions also initiated extensive sporulation. An isogenic relaxed (rel) strain produced little antibiotic activity, which decreased under partial amino acid deprivation. In rel+ cells, the addition of a low concentration of chloramphenicol, which reduces ppGpp synthesis, also reduced antibiotic synthesis in both normal and amino acid-starved bacteria, without appreciably affecting their growth rate. Guanosine starvation of a gua mutant initiated sporulation, but decreased antibiotic production. The results show that the stringent response initiates both sporulation (differentiation) and antibiotic production (secondary metabolism), but by different mechanisms. It appears that sporulation results from a decrease of GTP, whereas antibiotic synthesis results from a different effect of the stringent response.     

Rifampicin action on the ultrastructure of E. coli cells]

The ultrastructure of Coli bacteria exposed to rifampicin was studied. Dependence of the level of the ultrastructural changes on the antibiotic concentration and the time of incubation with the antibiotic was shown. After exclusion of the antibiotic from the medium the organism growth with normal ultrastructure was observed.     

EXPERIMENTS ON THE MOVEMENT OF STREPTOMYCIN IN CHERRY TREES

Systemic activity of streptomycin in cherry trees was investigated by studying the penetration of the antibiotic into leaves and its subsequent translocation.
Antibiotic penetration appreciably suppressed infection with Pseudomonas morsprunorum. In detached leaves the uptake of the antibiotic was progressive with time.
Widespread and rapid distribution of the antibiotic in the transpiration stream resulted from petiolar injection. Immersion of the laminae of intact leaves in the antibiotic resulted in a much slower translocation, possibly occurring in the transpiration stream.
Evidence suggests that the antibacterial activity detected in fruiting spurs after field spraying is not dependent on the presence of leaves, but results from direct penetration of the antibiotic through the dermal tissues of the spur.
It was concluded that foliar sprays at field concentrations were unlikely to result in significant systemic distribution of the antibiotic, but that localized activity might be an important factor in the control of bacterial canker.     

A new pigmented antibiotic from a soil streptomycete

An antibiotic designated Ass was isolated from a soil streptomycete -which showed wide antibacterial activity. The antibiotic was extracted and purified into a yellow powder. Its physico-chemical and antimicrobial properties indicated that it is a novel peptide antibiotic.     

Characterization of an antibiotic produced by Alteromonas luteoviolacea Gauthier 1982,85 isolated from Kinko Bay, Japan

An antibiotic produced by Alteromonas luteoviolacea strain 9K-V10 was recovered after cold acetone precipitation of culture supernatant fluids or lysates that had beenfrozen and thawed. The precipitate obtained from cell-free lysates was fractionated by DEAE ion-exchange chromatography. Further purification by gel-filtration chromatography yielded a single peak of antibiotic activity that corresponded to a protein peak with a molecular mass of approximately 100 kDa. After non-denaturing polyacrylamide gel electrophoresis, antibiotic activity co-migrated with a protein band. The isoelectric point of the antibiotic was estimated to be 7·7. Treatment of the concentrated active fraction with proteinase K or heating at 70°C for 10 min resulted in total loss of antibiotic activity. These results show that the antibiotic produced by Alt. luteoviolacea 9K-V10 is of a proteinaceous nature.     

Kinetics of Streptomyces baarnensis growth and antibiotic synthesis in batch and dialysis cultures

The kinetics of biomass and antibiotic formation in batch and dialysis culture of Streptomyces baarnensis at various initial concentrations limiting the substrate growth (glucose) has been studied. The antibiotic substances were synthesized by actively growing culture, its concentration in the cultural media was maximum in the log-phase. In continuous dialysis culture on the background of biomass lianer growth in the course of time the constant antibiotic concentration in the media proportional to the glucose input concentration has been established. The inactivation (decomposition) of antibiotic was immediately initiated after discontinuation of substrate supply and followed first kinetics order. Observed features were used for construction of kinetical model of antibiotic biosynthesis. A conclusion has been made that the dialysis culture gives opportunity for more effective antibiotic synthesis as compared with the batch one.     

Mutagenic effect of mitomycin C on different strains of oleandomycin producer Streptomyces antibioticus

Mitomycin C, a DNA-tropic antibiotic, was shown to have a lethal effect on spore sprouts of two strains of Streptomyces antibioticus, an organism producing oleandomycin. When the time of exposure to the antibiotic increased there was an almost equal decrease in the survival rate. The mutagen action on the morphological variation and antibiotic production of the two closely related strains were diverse due to their genetic differences. The strain isolated after the culture treatment with a chemical mutagen and subjected to a more prolonged maintaining selection showed lower variation with respect to its colony morphology. The other strain isolated after treatment of the culture with high concentrations of its own antibiotic showed lower variation with respect to its antibiotic production property. The shift in the antibiotic production in the direction of the low active variants was characteristic of the both highly productive strains.