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PubMed2006

Mutations conferring aminoglycoside and spectinomycin resistance in Borrelia burgdorferi.

We have isolated and characterized in vitro mutants of the Lyme disease agent Borrelia burgdorferi that are resistant to spectinomycin, kanamycin, gentamicin, or streptomycin, antibiotics that target the small subunit of the ribosome. 16S rRNA mutations A1185G and C1186U, homologous to Escherichia coli nucleotides A1191 and C1192, conferred >2,200-fold and 1,300-fold resistance to spectinomycin, respectively. A 16S rRNA A1402G mutation, homologous to E. coli A1408, conferred >90-fold resistance to kanamycin and >240-fold resistance to gentamicin. Two mutations were identified in the gene for ribosomal protein S12, at a site homologous to E. coli residue Lys-87, in mutants selected in streptomycin. Substitutions at codon 88, K88R and K88E, conferred 7-fold resistance and 10-fold resistance, respectively, to streptomycin on B. burgdorferi. The 16S rRNA A1185G and C1186U mutations, associated with spectinomycin resistance, appeared in a population of B. burgdorferi parental strain B31 at a high frequency of 6 x 10(-6). These spectinomycin-resistant mutants successfully competed with the wild-type strain during 100 generations of coculture in vitro. The aminoglycoside-resistant mutants appeared at a frequency of 3 x 10(-9) to 1 x10(-7) in a population and were unable to compete with wild-type strain B31 after 100 generations. This is the first description of mutations in the B. burgdorferi ribosome that confer resistance to antibiotics. These results have implications for the evolution of antibiotic resistance, because the 16S rRNA mutations conferring spectinomycin resistance have no significant fitness cost in vitro, and for the development of new selectable markers.

Aminoglycosides↗

PubMed1976

Sucrose-dependent spectinomycin-resistant mutants of Escherichia coli.

Spectinomycin-resistant (Spcr) mutants of Escherichia coli were isolated from nutrient agar plates containing 20% sucrose and 100 mug of spectinomycin per ml. About one-third of the Spcr mutants thus obtained were sucrose dependent (Sucd) and were classified into two types: I, those unable to grow on sucrose-free medium in the presence of spectinomycin; and II, those unable to grow on sucrose-free medium irrespective of the presence of spectinomycin. Most of these mutants were hypersensitive to antibiotics, dyes, and detergents and were abnormal in cell morphology, suggesting changes in cell envelopes. Reversion experiments indicated that the sucrose-dependent spectinomycin resistance and hypersensitivity to various chemicals were not independently induced properties. The Sucd-Spcr mutations of type I mutants were transducible by phage P1 and were mapped at the strA-aroE region.

Acriflavine↗

PubMed1977

Interaction of cytoplasmic membrane and ribosomes in Escherichia coli: spectinomycin-induced disappearance of membrane protein I-19.

Incubation of Escherichia coli with spectinomycin caused the disappearance of a major protein from the cytoplasmic membrane. This protein, called "I-19", was not a ribosomal protein. Its disappearance was not a result of the direct action of spectinomycin on the cytoplasmic membrane, but a result of its action on ribosomes. The disappearance was specifically induced by spectinomycin, and other antibiotics such as neomycin, erythromycin, and chloramphenicol had no effect. Although growth was not required for spectinomycin-induced disappearance of protein I-19 from the cytoplasmic membrane, the disappearance was not observed under conditions where protein synthesis was inhibited completely either by the addition of chloramphenicol or by cooling in ice. It is suggested that at least some ribosomes interact with the cytoplasmic membrane and that a modification of the mode of interaction through the action of spectinomycin on ribosomes caused the deletion of membrane protein I-19.

Bacterial Proteins↗

PubMed1984

Spectinomycin as initial treatment for gonorrhoea.

The prevalence of penicillinase producing Neisseria gonorrhoeae at this hospital increased exponentially from less than 0.5% in 1978 to 6.5% of all isolates in 1982. In January 1983 first line treatment for uncomplicated heterosexual anogenital gonorrhoea was therefore changed from ampicillin and probenecid to spectinomycin. This subsequently cured 95% of cases seen at the Praed Street Clinic. Although there was an initial fall in the monthly isolation rate of penicillinase producing N gonorrhoeae after the introduction of spectinomycin, this was not maintained. The exponential increase in the prevalence of the strain did slow in 1983, rising to only 8.7%. This, however, may have reflected a general decline in the rate of increase of penicillinase producing N gonorrhoeae throughout Britain. The failure to influence the prevalence of penicillinase producing N gonorrhoeae to any great degree may have been due in part to spectinomycin resistance in both penicillinase producing and non-penicillinase-producing N gonorrhoeae. All of the isolates appeared identical, apart from the presence of the 4.4 megadalton plasmid in penicillinase producing N gonorrhoeae, but they could not be linked epidemiologically. Changing treatment in only one of the many venereal diseases clinics in London, where patients have open access to all such clinics, is unlikely to affect the prevalence of penicillinase producing N gonorrhoeae. This has probably been more important than spectinomycin resistance in limiting the effectiveness of spectinomycin in reducing the prevalence of the strain.

Adolescent↗

PubMed1987

Plasmid profile of Neisseria gonorrhoeae in Nigeria and efficacy of spectinomycin in treating gonorrhoea.

The prevalence of penicillinase producing Neisseria gonorrhoeae (PPNG) strains has been steadily rising in Nigeria since 1979, and now about 80% of the strains of gonococci isolated in Ibadan are found to produce penicillinase. Spectinomycin has consequently become widely used in treating these infections. To ascertain the emergence of spectinomycin resistance, this study was undertaken to assess the in vivo susceptibilities of gonococcal strains to spectinomycin and other common antibiotics. Five hundred and twenty seven isolates were tested, of which 452 (85.5%) were PPNG strains. None of the strains were found to be resistant to 100 micrograms spectinomycin discs in vitro, whereas all 370 patients treated with the antibiotic were bacteriologically cured. Plasmid analysis shows that both "Asian" and "African" PPNG types are circulating in Nigeria. For the moment spectinomycin remains highly effective in treating gonococcal infections in west Africa.

Drug Resistance, Microbial↗

PubMed1980

The effects of spectinomycin and ethidium bromide on the synthesis of organelle rRNA and on ultrastructure in Ochromonas danica.

The effects of 24-h exposure to spectinomycin (100 microgram/ml) and ethidium bromide (1 microgram/ml) on the accumulation of chloroplast and mitochondrial rRNAs and on organelle ultrastructure were studied in greening cells of Ochromonas danica. Cells treated with ethidium bromide for 24 h divide at the same rate as controls but contain less than one third the normal amount of mitochondrial rRNA. Ultrastructural observations showed that these cells contain only 10% the number of mitochondrial ribosomes found in controls as well as fewer mitochondrial cristae. Ethidium bromide has no effect on chloroplast ultrastructure in Ochromonas. Greening cells treated with spectinomycin grow at close to control rates but contain 30-40% less chloroplast rRNA than do controls. Electron microscopy showed that spectinomycin disrupts the organization of chloroplast membranes and reduces the number of chloroplast ribosomes by 30%. Under these conditions, spectinomycin has no effect on mitochondrial rRNA or ultrastructure. Since spectinomycin is a specific inhibitor of translation on 70S ribosomes, these results are consistent with the possibility that at least some chloroplast ribosomal proteins are synthesized in the chloroplast of Ochromonas.

Cell Division↗

PubMed2003

The effects of lincomycin-spectinomycin and sulfamethoxazole-trimethoprim on hyaluronidase activities and sperm characteristics of rams.

The effects of lincomycin-spectinomycin and sulfamethoxazole-trimethoprim combinations on the hyaluronidase enzyme of serum and semen and on sperm characteristics in rams were determined. Thirthy-two Akkaraman rams were used. The rams were randomly divided into four groups. Group A and group B were determined as control groups of group C (lincomycin-spectinomycin) and D (sulfamethoxazole-trimethoprim), respectively. Combinations of lincomycin-spectinomycin and sulfamethoxazole-trimethoprim were administered at doses of 15 mg.kg(-1) intramuscularly and 12 mg.kg(-1) body weights orally, respectively. Blood and semen samples were collected at 4, 12, 24, 48, 72, 192 and 384 hr. Semen hyaluronidase activities of rams in group C increased significantly (p<0.001, <0.05) compared with the control group at 24 and 48 hr, respectively. Semen hyaluronidase activities in group D rams also increased significantly (p<0.001) in comparison with the control group at all times except 72 and 384 hr. Serum hyaluronidase activities increased significantly (p<0.01, <0.001) at 24 and 48 hr after treatment of lincomycin-spectinomycin. Additionally, significant (p<0.05, <0.001) increases were detected in the serum hyaluronidase activities of group D at 48 and 72 hr, respectively. No significant correlation was found between serum and semen hyaluronidase activities. Furthermore, significant increases (p<0.05) were observed in the percentages of motile sperm in the rams of group C and D compared with the control groups. The values of sperm concentration and total number of sperm in group C and D rams decreased significantly (p<0.001) in comparison with control groups. No significant correlations were found between the semen hyaluronidase activities and sperm characteristics. In conclusion, these findings show that the combinations of lincomycin-spectinomycin and sulfamethoxazole-trimethoprim do not have any harmful effects on hyaluronidase activities and sperm motility. However, the use of both antibiotic combinations in breeding rams during the ramming season is not advisable due to the decrease of sperm concentration.

Animals↗

PubMed1976

Drugs five years later. Spectinomycin.

A single intramuscular injection of 0.2 g of spectinomycin hydrochloride is highly effective for the treatment of uncomplicated anogenital infections with Neisseria gonorrhoeae. Spectinomycin hydrochloride is indicated for uncomplicated anogenital gonococcal infections in men and women who cannot receive penicillin or probenecid, and is the drug of choice for the retreatment of patients with uncomplicated anogenital gonococcal infection who have not responded to other antibiotics. A single dose of spectinomycin does not appear to be reliably effective in the treatment of gonococcal pharyngitis. Preliminary reports indicate that multiple-dose schedules of spectinomycin will prove to be effective in gonococcal pelvic inflammatory disease and disseminated gonococcal infection. In the dosage used for gonococcal infections, spectinomycin has little effect on infection with Treponema pallidum. Evaluation of this antibiotic against other microorganisms suggests little utility for this agent in conditions other than gonococcal infections.

Animals↗

PubMedDate not supplied

Application of microbial receptor assay to quantitation of residues of spectinomycin in bovine milk and comparison with liquid chromatographic analysis.

Spectinomycin-contaminated bovine milk samples were assayed by liquid chromatographic (LC) and microbial receptor methods. LC involved a newly developed analytical method to quantitate the concentration of spectinomycin in the contaminated milk samples. The receptor assay used reagents and the reaction system used for the Charm II spectinomycin assay. Three standard curves (selected range, full range, and second-order polynomial) were plotted for the receptor assay and used to quantitate spectinomycin in contaminated milk samples. The levels of spectinomycin obtained by the receptor assay, using only the standard curve in the selected range, were comparable to the results obtained by LC analysis.

Animals↗

PubMed1976

Spectinomycin - experimental investigations on possible side-effects.

Experimental investigations were performed to elaborate the immunogenic and anaphylactoid properties of spectinomycin. In guinea-pigs, no allergy against spectinomycin could be elicited. Furthermore, the absence of immunologic cross-reactions between spectinomycin and penicillin could be confirmed. The anaphylactoid activity of spectinomycin was found to be of very low order. No mast cell degranulating action could be observed. The data collected in this study permit the statement that - as far as acute unwanted actions of antibiotics are concerned - spectinomycin is an extremely safe drug.

Anaphylaxis↗

PubMed1974

Therapeutic effect of lincomycin and spectinomycin water medication on swine dysentery.

The therapeutic effects of various water medications on swine dysentery were determined in 223 pigs under controlled conditions. Carrier pigs were mixed with test animals until the disease was established. Lincomycin (22 mg/liter), spectinomycin (44 mg/liter) alone and lincomycin and spectinomycin in combination (66 mg/liter) and sodium arsanilate (161 mg/liter) in drinking water for seven days were the drugs evaluated. Negative and positive controls were also included. The experiment was terminated 41 to 43 days after initial medication. Mortality, mean value for stool consistency, incidence of dysenteric days and gross lesions of swine dysentery were the parameters measured for each treatment group.The lincomycin-spectinomycin water medication was effective for the treatment of swine dysentery. Pigs treated with lincomycin-spectinomycin had a higher survival rate, a lower incidence of dysenteric days and fewer gross lesions of swine dysentery than pigs treated with sodium arsanilate, lincomycin or spectinomycin alone or the infected controls (P < 0.05).

Animals↗

PubMed1982

[Spectinomycin therapy in combination with ampicillin in female patients with acute gonorrhea].

Clinical effects of spectinomycin (Trobicin, 2 g. i.m., once a day) and ampicillin (1 g/day, p.o., in 4 divided doses, for 2 days) given in combination on acute gonorrhea in female patients were studied, and sensitivity of isolated gonococci to each of these antibiotics was determined. 1. Clinical effects were evaluated by cultures of gonococci from discharge collected at 72 hours after spectinomycin administration and 12 hours after the last dose of ampicillin and both subjective and objective clinical findings. Among 20 cases of gonorrhea treated, 12 cases (60.0%) proved excellent in therapeutic results, 6 cases (30.0%) good, 1 case fair and 1 case produced no response. The success rate comprising excellent and good was 90.0%. The only side effect reported was pain of injection site in 1 case on 3 days after initiation of treatment, but no induration was noted. 2. MICs of spectinomycin and ampicillin for gonococci collected from the 20 cases were determined. Spectinomycin showed MICs of 6.25 approximately 12.5 micrograms/ml against 10(8) CFU/ml and 3.13 approximately 12.5 micrograms/ml against 10(8) CFU/ml. These values were below the blood concentration obtained at 6 hours after intramuscular administration of 2 g of spectinomycin. Ampicillin had MICs of 0.1 approximately 25 micrograms/ml against 10(8) CFU/ml and 0.1 approximately 3.13 micrograms/ml against 10(6) CFU/ml. Five strains, MICs of which were 6.25 approximately 25 micrograms/ml against 10(8) CFU/ml, were beta-lactamase-producing.

Acute Disease↗

PubMedDate not supplied

Effect of spectinomycin on Escherichia coli infection in 1-day-old ducklings.

Two challenge trials and one confirmation trial were conducted to evaluate the efficacy of spectinomycin in the treatment of 1-day-old ducklings infected with Escherichia coli. In the challenge trials, ducklings were injected in the right posterior thoracic air sac with 0.2 cm3 of broth containing 10(8) colony-forming units E. coli (strain O78, E38)/ml. Spectinomycin at dosage levels of 2.5 mg, 5.0 mg, and 10.0 mg of activity was injected subcutaneously 6 hours following infection. The confirmation trial was conducted to confirm the challenge trials; procedures were similar to those used in the challenge trials, except that only the 5.0 mg of activity dosage of spectinomycin was used. In both types of trials, spectinomycin-treated ducklings had significantly lower mortality and higher average weight gain, average daily gain, and feed consumption than infected unmedicated controls. These results indicate that spectinomycin is effective in treating ducks for experimentally induced colibacillosis caused by E. coli (strain O78, E38).

Animals↗

PubMedDate not supplied

Easy enzyme-linked immunosorbent assay for spectinomycin in chicken plasma.

An easy, sensitive, competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) for spectinomycin in chicken plasma was developed. Preparation of a spectinomycin-bovine serum albumin conjugate in which the hapten is linked to the carrier protein through the C-4 position is described. Antibodies raised against antigens in rabbits had excellent specificity for spectinomycin, exhibiting a cross-reactivity of 44.0% with dihydrospectinomycin and 13.8% with tetrahydrospectinomycin. No cross-reactivity was observed with other antibiotics. The detection limit of the CI-ELISA was 2 ng/mL (equivalent into 40 ng/mL undiluted chicken plasma) spectinomycin. Known amounts (0.1-100 micrograms/mL) of spectinomycin were added to chicken plasma and then analyzed. Average recoveries were 97-110%. This procedure may be used without prior extraction of samples.

Animals↗

PubMed1991

Mutation proximal to the tRNA binding region of the Nicotiana plastid 16S rRNA confers resistance to spectinomycin.

Nicotiana tabacum lines carrying maternally inherited resistance to spectinomycin were obtained by selection for green callus in cultures bleached by spectinomycin. Two levels of resistance was found. SPC1 and SPC2 seedlings are resistant to high levels (500 micrograms/ml), SPC23 seedlings are resistant to low levels (50 micrograms/ml) of spectinomycin. Lines SPC2 and SPC23 are derivatives of the SR1 streptomycin-resistant plastome mutant. Spectinomycin resistance is due to mutations in the plastid 16S ribosomal RNA: SPC1, an A to C change at position 1138; SPC2, a C to U change at position 1139; SPC23, a G to A change at position 1333. Mutations similar to those in the SPC1 and SPC2 lines have been previously described, and disrupt a conserved 16S ribosomal RNA stem structure. The mutation in the SPC23 line is the first reported case of a mutation close to the region of the 16S rRNA involved in the formation of the initiation complex. The new mutants provide markers for selecting plastid transformants.

Chloroplasts↗

PubMed1994

Mutations conferring lincomycin, spectinomycin, and streptomycin resistance in Solanum nigrum are located in three different chloroplast genes.

A number of Solanum nigrum mutants resistant to the antibiotics spectinomycin, streptomycin and lincomycin have been isolated from regenerating leaf strips after mutagenesis with nitroso-methylurea. Selection of streptomycin- and spectinomycin-resistant mutants has been described earlier. Lincomycin-resistant mutants show resistance to higher levels of the antibiotic than used in the initial selection, and in the most resistant mutant (L17A1) maternal inheritance of the trait was demonstrated. The lincomycin-resistant mutant L17A1 and a streptomycin plus spectinomycin resistant double mutant (StSp1) were chosen for detailed molecular characterisation. Regions of the plastid DNA, within the genes encoding 16S and 23S rRNA and rps12 (3') were sequenced. For spectinomycin and lincomycin resistance, base changes identical to those in similar Nicotiana mutants were identified. Streptomycin resistance is associated with an A-->C change at codon 87 of rps12 (converting a lysine into a glutamine), three codons upstream from a mutation earlier reported for Nicotiana. This site has not previously been implicated in streptomycin resistance mutations of higher plants, but has been found in Escherichia coli. The value of these mutants for studies on plastid genetics is discussed.

Base Sequence↗

PubMed2006

Determination of spectinomycin hydrochloride and its related substances by HPLC-ELSD and HPLC-MSn.

A new and simple high-performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) method for the determination of spectinomycin hydrochloride and its related substances was developed. The column was Agilent SB-C(18) (250 mm x 4.6 mm, 5 microm). The mobile phase was 25 mM trifluoroacetic acid. The drift tube temperature was 40 degrees C. The pressure of nebulizing gas was 3.5 bar. Good separation of spectinomycin from main related substances could be achieved. The standard curve was rectilinear in the range of 0.07-3.8 mg/ml (r = 0.9997). Precision was 1.0% (R.S.D.). The limit of detection was 6 microg/ml. The method is simple and rapid, and the results are accurate and reproducible. The HPLC-MS(n) method was used to characterize the structures of impurities contained in the spectinomycin. In positive mode, impurities were elucidated by use of electrospray ion trap mass spectrometry in the multi-stage MS full scan mode. The possible structures of impurities C and D in spectinomycin were deduced based on the HPLC-MS(n) data.

Anti-Bacterial Agents↗

PubMed1998

Quantitation of spectinomycin residues in bovine tissues by ion-exchange high-performance liquid chromatography with post-column derivatization and confirmation by reversed-phase high-performance liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry.

Determinative and confirmatory methods of analysis for spectinomycin residue in bovine kidney, liver, muscle and fat have been developed. The determinative method is a single-column HPLC ion-exchange procedure that incorporates a two-step post-column oxidation of the secondary amines to primary amines followed by derivatization with o-phthalaldehyde. The method was validated in all tissues to a low-end concentration of 0.10 micrograms/g (limit of quantitation) and to a high-end of 10 micrograms/g for kidney, which is the rate-limiting tissue for residues of spectinomycin. The recovery of spectinomycin from all tissues was > 80% and the variability (R.S.D.) was generally < 10%. For liver, an alternative reversed-phase HPLC separation was required for incurred-residue samples. The confirmatory method employed an atmospheric pressure chemical ionization-MS-MS approach utilizing a rapid reversed-phase HPLC system with a mobile phase of methanol and 1% acetic acid. The protonated molecular ion for spectinomycin at m/z 333 produced four diagnostic reaction-product ions at 98, 116, 158 and 189 for confirmation. The method was validated to a lower limit of confirmation of 0.10 micrograms/g.

Animals↗