Lincomycin at Subinhibitory Concentrations Potentiates Secondary Metabolite Production by Streptomyces spp.

Antibiotics have either bactericidal or bacteriostatic activity. However, they also induce considerable gene expression in bacteria when used at subinhibitory concentrations (below the MIC). We found that lincomycin, which inhibits protein synthesis by binding to the ribosomes of Gram-positive bacteria, was effective for inducing the expression of genes involved in secondary metabolism in Streptomyces strains when added to medium at subinhibitory concentrations. In Streptomyces coelicolor A3(2), lincomycin at 1/10 of its MIC markedly increased the expression of the pathway-specific regulatory gene actII-ORF4 in the blue-pigmented antibiotic actinorhodin (ACT) biosynthetic gene cluster, which resulted in ACT overproduction. Intriguingly, S. lividans 1326 grown in the presence of lincomycin at a subinhibitory concentration (1/12 or 1/3 of its MIC) produced abundant antibacterial compounds that were not detected in cells grown in lincomycin-free medium. Bioassay and mass spectrometry analysis revealed that some antibacterial compounds were novel congeners of calcium-dependent antibiotics. Our results indicate that lincomycin at subinhibitory concentrations potentiates the production of secondary metabolites in Streptomyces strains and suggest that activating these strains by utilizing the dose-response effects of lincomycin could be used to effectively induce the production of cryptic secondary metabolites. In addition to these findings, we also report that lincomycin used at concentrations for markedly increased ACT production resulted in alteration of the cytoplasmic protein (F_oF₁ ATP synthase α and β subunits, etc.) profile and increased intracellular ATP levels. A fundamental mechanism for these unique phenomena is also discussed.     

Lincomycin,clindamycin and their applications

Lincomycin and clindamycin are lincosamide antibiotics used in clinical practice. Both antibiotics are bacteriostatic and inhibit protein synthesis in sensitive bacteria. They may even be bactericidal at the higher concentrations that can be reached in vivo . Clindamycin is usually more active than lincomycin in the treatment of bacterial infections, in particular those caused by anaerobic species; and it can also be used for the treatment of important protozoal diseases, e.g. malaria, most effectively in combination with primaquine. Resistance to lincomycin and clindamycin may be caused by methylation of 23S ribosomal RNA, modification of the antibiotics by specific enzymes or active efflux from the periplasmic space.     

Further similarities between chloroplast and bacterial ribosomes

Summary Protein synthesis by chloroplasts isolated under aseptic conditions from Phaseolus vulgaris leaves is inhibited by the bacterial antibiotics spectinomycin, lincomycin, and erythromycin; that by chloroplasts from Nicotiana tabacum leaves is inhibited by spectinomycin and lincomycin but not by erythromycin. Protein synthesis by cytoplasmic ribosomes from plants and animals is not inhibited by these compounds, nor is amino acid activation by the soluble fraction from bean chloroplasts. These results suggest that chloroplast ribosomes possess sites which bind several unrelated bacterial antibiotics and support the idea that chloroplasts originated from prokaryotic cells. These antibiotics may be useful in studying the process of chloroplast formation in intact cells.     

Antibiotic sensitivity of plague microbe strains from foreign countries]

The method of serial dilutions on the Hottinger agar was applied to comparative assay of antibiotic sensitivity in 50 strains of the plague microbe isolated abroad and in 5 strains isolated in the plague focus in the Central Caucasus. The antibiotics used in the assay were the following: streptomycin, gentamicin, doxycycline, monomycin, kanamycin, tetracycline, erythromycin, ristomycin, lincomycin and polymyxin M. Irrespective of the origin, all the isolates were resistant to erythromycin, lincomycin and polymyxin M. The levels of the sensitivity to the other antibiotics were different. The data serve as a ground for the statement that there is no tendency to development of antibiotic resistance in the plague microbe in patients treated with high doses of the antibiotics and mainly streptomycin. Along with streptomycin, such antibiotics as gentamicin, tetracycline, doxycycline and kanamycin are useful in the therapy of plague and require further investigation.     

Results of 10 years of use of lincomycin (1966-1976) in the clinics of the N. N. Priorov Central Research Institute of Traumatology and Orthopedics]

During 10 years 1063 patients were treated with lincomycin used parentally or orally at the N. N. Priorov Central Research Institute of Traumatology and Orthopedy. The doses and the rate of its use depended on the state of the patient, its age and weight. Lincomycin was used for the treatment of patients with osteomyelitis or purulent wound infection, as well as for prophylaxis of suppuration. The drug was used for a long period of time under conditions of the same hospital, and it was shown that it remained up to the present days highly effective in therapy of infections and especially bone infections caused by staphylococci sensitive to it. The 10-year study of staphylococcal sensitivity to lincomycin revealed an insignificant increase in the development of resistance to it. The paper presents data on the importance of adequate surgical interventions in addition to the antibiotic therapy in cases with bone infections. A possibility of lincomycin combined use with other antibiotics and gentamicin or kanamycin in particular was shown. Complications, such as diarrhea and urticaria were registered in 11 patients.     

Effect of antibiotic and vaccine therapies on the succinate dehydrogenase and phosphatase activity of liver cells of mice infected with Staphylococcus

Changes in the activity of succinate dehydrogenase (SDH), total and acid phosphatase (TP and AP) were studied in treatment of laboratory animals with rifampicin, lincomycin and with inactivated staphylococcal vaccine used alone or in combinations. It was shown that immunization of the animals with inactivated staphylococcal vaccine under conditions of experimental staphylococcal infection promoted stimulation of the enzyme activity. Rifampicin and lincomycin used for the treatment of such animals lowered the activity of the enzymes. The suppressing effect of the antibiotics increased with an increase in the period of their use. It should be noted that the inhibitory effect of rifampicin on the activity of SDH, TP and AP was less pronounced than that of lincomycin. The combined use of the vaccine and antibiotics for the treatment of the animals promoted an increase in the enzyme activity as compared to the use of the antibiotics alone. Sometimes the activity of SDH, TP and AP reached the control levels in such animals or the levels observed in the animals treated with the vaccine alone. Stimulation of the enzyme activity was more pronounced when the vaccine was used in combination with rifampicin.     

Effect of antibiotic therapy and vaccine therapy on the phagocytic reaction in mice infected with Staphylococcus

The time course of changes in the activity, intensity and completeness of phagocytosis with leukocytes of the peritoneal exudate was studied on mice with experimental staphylococcal infection treated with rifampicin, lincomycin and inactivated staphylococcal vaccine used alone or in combination. It was shown that immunization of the animals with inactivated staphylococcal vaccine promoted stimulation of the phagocytic defense. Rifampicin and lincomycin applied therapeutically induced a decrease in the activity, intensity and completeness of phagocytosis. It should be noted that rifampicin had a less pronounced inhibitory effect than lincomycin. The combined use of vaccine and antibiotics with therapeutic purposes promoted an increase in phagocytosis as compared to the use of the antibiotics alone. The combined therapy sometimes resulted in completeness of phagocytosis making it reach the control values (the 10th and 15th days, rifampicin and vaccine). It should be noted that a more pronounced stimulation of the activity, intensity and completeness of the phagocytosis was observed with the use of the combination of rifampicin and the vaccine.     

Antibiotic sensitivity of Staphylococci populating the breast skin of nursing women

Sensitivity to penicillins, tetracycline, erythromycin and lincomycin was tested in 1513 staphylococcal strains isolated from the skin of various anatomic areas of the breast of nursing mothers. It was shown that 82.9, 78.4 and 33.9 per cent of the isolated cultures were highly sensitive to lincomycin, erythromycin and methicillin respectively. Sensitivity to tetracycline and benzylpenicillin was detected in 33.1 and 19.8 per cent of the cultures respectively. The study of the resistance spectra of 1343 strains resistant to certain antibiotics revealed that 498 cultures (37.1 per cent) were polyresistant. None of the tested antibiotics could be used for selective decontamination of the breast skin with respect to S. aureus in prophylaxis of lactic mastitis.     

Larval Apis mellifera L. (Hymenoptera: Apidae) mortality after topical application of antibiotics and dusts

Beekeepers apply various dusts to honey bee, Apis mellifera L., colonies to dislodge parasitic mites and control bacterial brood diseases. Anecdotal reports by beekeepers indicate that the antibiotic oxytetracycline (OTC) can be toxic when applied in powdered sugar to cells containing immature bee brood, but it was not known whether the purported toxicity is caused by the antibiotic or the sugar carrier. Additionally, the toxicity of various dusts, proposed for parasitic mite control, is poorly known. In the current studies, we tested OTC and two other antibiotics (tylosin and lincomycin, candidate compounds for use in honey bee colonies) in a powdered sugar carrier for larval toxicity. We also tested for larval toxicity, several dusts that have been proposed for mite control. OTC caused significant brood mortality of approximately 80% at the concentrations used in the hive (200 mg in 20 g sugar). In contrast, tylosin and lincomycin at the 200 mg dose were both similar to untreated controls, and only five times that concentration (1000 mg) caused significant brood mortality of approximately 65%. The addition of dusts, wheat flour, talc, and a commercially available protein supplement, BeePro, resulted in mortality levels between 65 and 80%, similar to that seen with OTC. The common antibiotic carrier, powered confectioners sugar, was nontoxic. The use of 100 unsealed brood cells was demonstrated to be a reliable means of assessing potential adverse affects of dry compounds on larval honey bees. Two new candidate antibiotics for use in honey bee colonies were less toxic to larval bees than the currently labeled antibiotic, OTC.     

Sensitivity of the wound microflora of oncological patients to some new antibiotics]

Sensitivity of the microflora of the oncological patients' wounds to the new antibiotics, such as gentamicin, kanamycin, oxacillin, ampicillin and lincomycin was studied with the help of the disc method. The discs with the above antibiotics were prepared under laboratory conditions in accordance with the respective instructions in the WHO. Sensitivity of 429 bacterial cultures, including 98 cultures of pathogenic staphylocci, 45 cultures of Enterococci, 43 hemolytic streptococci, 143 cultures of Escherichia, 50 cultures of Ps. aeruginosa and 50 cultures of Proteus was determined. The studies showed that gentamicin was the most active antibiotic aganist all the microbial species isolated from the surgical and other wounds of oncological patients. It may be used in treatment of the infections caused by association of the microbes belonging to different species, as well as in treatment of purulent processes before elucidating their etiology, 16.7 per cent of the Enterococcal isolates were resistant to gentamicin. Monomycin, kanamycin, oxacillin, lincomycin and macrolide antibiotics at present are sufficiently active against pathogenic staphylococci and hemolytic streptococci.     

Characterization of the neuromuscular block produced by clindamycin and lincomycin.

The site of neuromuscular blockade induced by clindamycin and lincomycin was studied on isolated nerve and nerve-muscle preparations. Clindamycin (3.6 X 10(-3) M) but not lincomycin (up to 1.5 X 10(-2) M) had a local anaesthetic effect on a frog desheathed nerve preparation. Clindamycin (8 X 10(-4) M) and lincomycin (4 X 10(-3) M) depressed the response of the rat diaphragm to nerve stimulation and to direct muscle stimulation in parallel. This indicated that the predominant neuromuscular blocking effect of these antibiotics was due to an effect on the muscle. Clindamycin was fivefold more potent than lincomycin in this effect, and the unionized form of both drugs was the active form. Lincomycin (4 X 10(-3) M) but not clindamycin (8 X 10(-4) M) also had some depressant effect on nerve-muscle transmission as indicated by the interaction of the effects of the antibiotics and d-tubocurarine. The significance of these findings is discussed in relation to the acute clinical toxicity of these antibiotics.     

Sensitivity of 56 strains of Streptococcus group B to 12 antibiotics: its determination by dilution and diffusion in agar

Sensitivity of 56 streptococcal strains of group B to 12 antibiotics was studied with the method of dilution in a special solid medium for cultivation of streptococci and with the method of agar diffusion in the same medium. All the strains were found to be sensitive to chloramphenicol, ristomycin and erythromycin. The predominating majority of the strains were sensitive to beta-lactam antibiotics and lincomycin. All the strains were resistant to aminoglycoside antibiotics, such as streptomycin and gentamicin. More than 85 per cent of the strains were resistant to tetracycline. Strains with multiple resistance to 2-7 antibiotics were detected. Satisfactory correlation between the two methods was observed. It was shown to be clinically advisable to determine the sensitivity of streptococci of group B to beta-lactam antibiotics, erythromycin and lincomycin.     

Effect of peptide antibacterial factor and changes in Staphylococcus susceptibility to antibiotics]

The susceptibility to antibiotics of the Staphylococcus epidermidis cells in the presence of low-molecular weight peptide factor was investigated. The factor was isolated from the Styaphylococcus warneri culture media. The factor enhanced the S. epidermidis susceptibility to ampicillin, chloramphenicol, lincomycin, rifampicin, tetracycline, cefalexine, erythromycin and fusidine. The same effect was demonstrated for the cells of S. epidermidis resistant to cadmium ions along with resistance to several antibiotics. However the cells reaction in this case was expressed to a lower extent. It was suggested that revealed alterations in staphylococci susceptibility to antibiotics was due to the peptide factor effect on cytoplasmic membrane permeability that resulted in nonspecific increase in accumulation of various compounds with low-molecular weight.     

Crystal and molecular structure and absolute configuration of lincomycin hydrochloride monohydrate

Lincomycin is a broad-spectrum antibiotic synthesized by Streptomyces lincolnensis that is particularly active against Gram-positive bacteria. It is widely used in human and veterinary applications. The crystal structure of lincomycin has been undertaken with a view to obtain the conformational and structural features of the drug in order to afford a comparison of its structural features with other aminoglycoside antibiotics. We report here the details of its structural and conformational features as determined by single-crystal X-ray crystallography. Crystals of lincomycin hydrochloride are orthorhombic, space group P2(1)2(1)2, with the cell dimensions a=18.5294(3) Angstroms, b=20.5980(4) Angstroms, c=6.17380(10) Angstroms, V=2356.35(7) Angstroms3. The structure was solved using X-ray diffraction data and refined to a final R-value of 0.0391 for 2321 reflections (I > or = 2sigma). The absolute configuration was established using the anomalous dispersion of the sulfur and chlorine atoms in the structure. The molecule consists of an amino acid linked by an amide group to a monosaccharide of galactose stereochemistry. A network of hydrogen-bonds stabilizes the crystal structure.     

Various methods of using lincomycin for treating acute and chronic hematogenic osteomyelitis in children]

Comparative data on the treatment of 209 children with acute and chronic hematogenic osteomyelitis are presented; 128 patients hospitalized before 1974 were treated with antibiotics, mainly penicillin and streptomycin without sensitivity testing. From 1974 81 children were treated with lincomycin; 80 per cent of the isolates were sensitive to this antibiotic. In lincomycin therapy the method of electrophoresis on the disease focus, intrabone administration of the drug and administration of the drug into the bone cavity together with the blood clot during surgical interventions in cases with chronic hematogenic osteomyelitis were used. A marked decrease in the rate of the chronic forms of the disease was registered (from 77.2 to 8.8 per cent).     

Peptidyl puromycin synthesis; effect of several antibiotics which act on 50 S ribosomal subunits

The effects of several antibiotics which are know to bind with 50 S ribosomal subunits, on the formation of several di- and tri-peptidyl puromycins have been examined. Tylosin and spiramycin inhibited the formation of phenylalanyl-(14)C-phenylalanyl-puromycin, glycyl-(14)C-phenyllalanyl-puromycin, leucyl-(14)C-phenylalanyl-puromycin, N(epsilon)-carbobenzoxylysyl-(14)C-phenylalanyl-puromycin, and valyl-glycyl-(14)C-phenylalanyl-puromycin as well as N-acetyl-(14)C-phenylalanyl-puromycin. Of these compounds, erythromycin and oleandomycin selectively inhibited the formation of phenylalanyl-(14)C-phenylalanyl-puromycin. Although chloramphenicol and lincomycin inhibited the formation of most of these peptidyl puromycins, the formation of phenylalanyl-(14)C-phenylalanyl-puromycin and leucyl-(14)C-phenylalanyl-puromycin was found to be resistant to these antibiotics. So far, no significant effect of siomycin has been observed on pepetidyl puromycin formation in the absence of G factor.     

Sensitivity of Mycoplasma hominis cultures isolated from clinical material to certain antibiotics]

Sensitivity to II antibiotics of 80 strains of M. hominis isolated from patients with various inflammatory processes of the urogenital tract was studied by the method of suppressing the metabolic activity. Inhibition of the arginine metabolism of mycoplasma was used as a test for determination of the growth suppression. All the strains tested were highly sensitive to tetracycline and lincomycin. Kanamycin and neomycin were less active against M. hominis. All the strains tested were resistant to erythromycin, oleandomycin, ristomycin, novobiocin and streptomycin. The inhibitory effect of tetracycline and lincomycin on M. hominis decreased by the 5th day.     

一株林可霉素降解菌的筛选鉴定及其降解机制

【背景】抗生素污染越来越引起人们的关注。利用微生物处理抗生素污染被认为是一种环境友好型的方法。【目的】筛选林可霉素高效降解菌并研究其降解机制。【方法】经形态学观察、生理生化鉴定和16S rRNA基因测序分析进行鉴定;通过PCR技术和质谱分析技术对该菌抗性基因和降解产物等进行分析。【结果】从林可霉素菌渣堆肥样本中获得一株高效降解林可霉素的假单胞菌(Pseudomonas RST-1),该菌在林可霉素浓度为3.0 g/L的牛肉膏蛋白胨培养基上培养40 h后,林可霉素降解率高达57.3%。该菌含有intI1、sul1、sul2等抗性基因,降解产物为去甲基林可霉素和2-丙基-N-甲基脯氨酸。【结论】菌株RST-1具有高效降解林可霉素的能力,推测可能的降解机制为去甲基化和酰胺键水解作用,该菌株降解特性及降解机制研究为林可霉素降解工程菌及其高效降解菌剂的研制奠定了基础。     

Therapeutic effect of some antibiotics on experimental staphylococcal infection and its correlation with in vitro activity of antibiotics in sub-inhibitory concentration against Staphylococcus aureus strains

The aim of the study was to demonstrate of whether the therapeutic effects of antibiotics depend on their in vitro activity in sub-inhibitory concentrations against staphylococci. Cloxacillin, gentamicin and lincomycin were used in the study. Groups of S. aureus strains, containing 6 strains with similar MIC values each but different sensitivity to sub-inhibitory antibiotic concentrations (sub-MIC) were selected (a total of 36 trains): i. strains increasing their sensitivity to phagocytosis and bactericidal activity of rabbit leukocytes after incubation with an antibiotic in 0.1 MIC concentration, ii. strains with sensitivity to the above factors unaffected by incubation with an antibiotic in 0.5 MIC concentration. The doses of staphylococci causing death of 90-100% of Swiss albino mice 10 days after i.p. infection were determined. The injected doses (LD 90-100) and various doses of antibiotics were used to determine ED50 values as well as the survival rate of the mice with experimental staphylococcal infections after treatment with these antibiotics. It was demonstrated that effective doses (ED 50) of the antiboitics were significantly lower when the antibiotics were administered once to mice infected with strains S. aureus sensitive to sub-MIC concentrations of the investigated antibiotics than for mice infected with strains resistant to their sub-MIC concentrations. Similar correlations were observed in mice which were given the antibiotics several times (for 7 days): the percentage of the surviving mice was higher in the group infected with sub-MIC sensitive strains. The therapeutic effect of cloxacillin, gentamicin and lincomycin demonstrated a significant correlation with the S. aureus strains used to induce the infections and their sensitivity, or lack of sensitivity in vitro, to phagocytosis and bactericdal activity of leukocytes in the presence of antibiotics in sub-MIC concentrations.     

Method of oblique illumination in studying the morphology of Staphylococcus aureus colonies sensitive and resistant to antibiotics]

The morphology of the colonies of Staph. aureus sensitive and resistant to antibiotics and the colonies with artificial resistance to benzylpenicillin, ampicillin, oxacillin, methicillin, cephaloridin and lincomycin was studied with the method of slanting light. Changes in the morphology of the colonies were most pronounced at the beginning of the microbial resistance development first of all in the cultures of Staph. aureus resistant to cephaloridin and lincomycin.