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Moellerella wisconsensis: identification, natural antibiotic susceptibility and its dependency on the medium applied.

The present study establishes a data compilation on biochemical features and natural antibiotic susceptibilities of Moellerella wisconsensis strains. 17 moellerellae isolated from humans (n = 11), food (n = 5) and water (n = 1) were tested. Identification was carried out using two commercially available systems and conventional tests. MIC determinations of 74 antibiotics were performed applying a microdilution procedure in Cation-adjusted Mueller Hinton broth and IsoSensitest broth. M. wisconsensis was naturally sensitive to doxycycline, minocycline, all tested aminoglycosides, numerous beta-lactams, all fluoroquinolones, folate-pathway inhibitors, chloramphenicol and nitrofurantoin. Natural resistance was found with oxacillin, penicillin G, all tested macrolides, lincomycin, streptogramins, ketolides, glycopeptides, fusidic acid, linezolid and rifampicin. Medium-dependent differences in susceptibility affecting clinical assessment criteria were seen with tetracycline, clindamycin and fosfomycin. From the data of the present study it is possible that some moellerellae are misidentified as Klebsiella pneumoniae subsp. ozaenae.

Anti-Bacterial Agents↗

Feed additives and contaminants as a cause of equine disease.

The equine practitioner often encounters serious diagnostic and therapeutic challenges regarding the specific origin of a disease. Such challenges may occur when horses become unaccountably ill after consuming what was thought to be acceptable feed but which in fact was contaminated or contained additives intended for other species. Examples of such additives and contaminants are monensin, lasalocid, salinomycin, blister beetles, and such antimicrobics as lincomycin and clindamycin.

Animal Feed↗

Phenotypic and genetic diversity among Bacillus sphaericus strains isolated in Brazil, potentially useful as biological control agents against mosquito larvae.

Thirty mosquitocidal strains of Bacillus sphaericus isolated from different sources and localities in Brazil were characterized phenotypically and genetically to determine their relationship. Among the strains tested, 93.3% were shown to be resistant to lincomycin, 96.6% to novobiocin, 60% to chloramphenicol and all strains were resistant to streptomycin. Resistance to HgCl2, NiSO4.6H2O and CuSO4 was observed in 83.3, 86.6 and 100% of the strains, respectively. All strains were inhibited by the presence of CoSO4. Tolerance to ethanol and variable responses to different amounts of creolin, phenol and xylol was also observed. Amplification of DNA of each of 30 isolates using repetitive primers allowed the identification of 5 groups of similar strains in BOX-PCR and 8 groups in REP-PCR. Using cloned toxin genes from B. sphaericus as probes in hybridization studies, 83% of the strains studied hybridized to the bin probe and 90% to the mtx probe. A comparison of the 30 strains by similarity matrix analysis using the data obtained in all approaches used in this study resulted in 22 groups (16 groups among the 24 high-toxicity strains) at 100% similarity, indicating a high degree of diversity among the strains tested. Some of the strains studied here, which are resistant to different stress conditions, should be considered for further ecological studies.

Animals↗

Antimicrobial susceptibility of potentially pathogenic halophilic vibrios isolated from seafood.

Susceptibility patterns to 27 antimicrobial agents and beta-lactamase production were investigated in potentially pathogenic halophilic vibrios from seafood. The effect of salinity on the response to the drugs in vitro was also studied. All isolates were uniformly sensitive to choramphenicol, imipenem, meropenem but resistant to lincomycin. All were highly sensitive to oxolinic acid, trimethoprim-sulphamethoxazole, doxycycline, flumequine, cefotaxime, nalidixic acid and ciprofloxacin. Some strains of V. harveyi, V. alginolyticus and V. parahaemolyticus apparently had mechanisms of resistance to several beta-lactam antibiotics other than by the production of beta-lactamases. Sixty-nine strains produced penicillinase but a low correlation between beta-lactamase activity and resistance to beta-lactam antibiotics was noted. The salt concentration affected the in vitro susceptibility of halophilic vibrios and the effect of salinity depended on both the individual strains and the antimicrobial tested.

Animals↗

Antimicrobial susceptibility of Streptococcus species isolated from clinical mastitis in dairy cows.

The antimicrobial susceptibility was determined for 50 Streptococcus uberis, 42 S. dysgalactiae subsp. dysgalactiae and eight S. agalactiae strains isolated from cow mastitis. Only 27% of the strains were susceptible to all antimicrobial compounds tested. Resistance to tetracycline was most frequent (particularly for S. dysgalactiae strains), then macrolide and/or lincomycin resistance. High level resistance to streptomycin and kanamycin was detected. All S. dysgalactiae and S. agalactiae strains were susceptible to beta-lactams but 44% of the S. uberis strains showed an elevated penicillin G MIC. All strains were susceptible to chloramphenicol and rifampicin.

Animals↗

Antimicrobial susceptibility testing of Actinomyces pyogenes: comparison of disk diffusion test and Api ATB Strep system with the agar dilution method.

The use of the disk diffusion and the Api ATB Strep system or a related technique for a few antibiotics was compared with the agar dilution method for antimicrobial susceptibility testing of 103 clinical isolates of Actinomyces pyogenes. There was complete agreement between disk diffusion and MIC in 98.8%, with minor errors, in 0.7% and with very large errors, in 0.5% of cases. The common criteria for the interpretation of zones could be kept for penicillin G, amoxicillin, methicillin, cephalothin, erythromycin, lincomycin, pristinamycin, vancomycin, rifampin, streptomycin, kanamycin, gentamicin and chloramphenicol. The proposed breakpoints for a tetracycline 30 IU disk were > or = 22 mm for susceptibility and < 22 mm for resistance. There was complete agreement between the ATB strip system and MIC in 98.3% of cases, with minor errors in 0.9% and with very large errors, in 0.8% of cases. All the dissimilarities were due to streptomycin and tetracycline and results could not be reported for these 2 antibiotics when using this system. Wells with kanamycin and gentamicin at low concentrations as well as chloramphenicol should be included by the manufacturer.

Actinomyces↗

Pneumococcal carriage amongst Australian aborigines in Alice Springs, Northern Territory.

In Alice Springs and its vicinity, a single nasal swab was collected from 282 Australian aborigines in May 1981 to determine nasal carriage rates of pneumococci. Each swab was inoculated on blood agar and on gentamicin blood agar. The carriage rates were 89% in children, 39% in adolescents and 34% in adults. In all, 27 serotypes of pneumococci were met with and 15 (4%) of subjects yielded two or more serotypes. In children, types 23, 19, 6, 22 and 6 were predominant (in that order), whereas type 3 was commonest in older subjects. Approximately 25% children and 5% adults yielded drug-insensitive pneumococci. Resistance to benzylpenicillin, tetracycline and co-trimoxazole was met with, resistant pneumococci showed five resistance patterns and belonged to nine serotypes, predominantly types 19 and 23. All isolates were sensitive to chloramphenicol, erythromycin, lincomycin and rifampicin. The carriage rate of drug-insensitive pneumococci was 100-fold higher amongst children sampled than in non-aboriginal children in Australia.

Adolescent↗

A community outbreak of group A beta haemolytic streptococci with transferable resistance to erythromycin.

Erythromycin resistance amongst group A streptococci (GAS) in Great Britain is a relatively rare occurrence and outbreaks have been sporadically reported. Over an 8-month period in 1986 ten associated cases occurred in the town of Bridgewater in Somerset. Isolates were group A, type M4 and resistant to erythromycin (MIC 8 mg/l) but sensitive to lincomycin and clindamycin. Erythromycin resistance was transferable from all isolates to a group A recipient strain. No plasmid DNA could be detected in the original isolates or transconjugants.

Adult↗

Pneumococcal carriage amongst children in Adelaide, South Australia.

Amongst 1267 healthy children 6 months to 4.5 years of age in Adelaide, the pneumococcal carriage rate from a single nasal swab sampling was 29% in the period 1980-1. Of 269 children, sampled monthly on five occasions, 91% carried a pneumococcus on one or more occasions: 55% carried a single type, 33% carried two types, 2% carried three types and 1% carried four types; 18% carried a pneumococcus on either 4 or 5 occasions. The commonest types encountered were types 6, 19 and 23 in that order, and these three types constituted 57% of the total: other common types (greater than 5% of the total) were types 14, 15 and 11, and the six commonest types constituted 77% of the total. Of these, types 6, 14, 19 and 23 commonly cause systemic disease in children; on the other hand types 11 and 15 cause disease infrequently. The number of strains showing antimicrobial drug resistance was low: on quantitative testing 0.7% of 291 isolates examined showed relative resistance to benzylpenicillin and 0.7% were resistant to tetracycline; 10.9% of 230 isolates examined showed resistance to co-trimoxazole; dual or multiple drug resistance was not detected, and all isolates tested were susceptible to chloramphenicol, erythromycin, lincomycin and rifampicin.

Carrier State↗

Systemic pneumococcal disease in Norway 1995-2001: capsular serotypes and antimicrobial resistance.

A total of 4624 pneumococcal isolates from episodes of systemic pneumococcal disease were received at the Norwegian Institute of Public Health during the period 1995-2001. All isolates were serotyped and tested for susceptibility to benzylpenicillin, lincomycin, erythromycin, tetracycline and trimethroprim sulphamethoxazole. The proportion of strains resistant to these antimicrobial agents remained stable at a low level, ranging from 0.1% for benzylpenicillin to 2.5% for erythromycin. The distribution of serotypes was also stable over the 7 years: serotypes 1, 4, 9, 14, 7, 6 and 23 were the most frequent, representing 70.5% of isolates. Overall, 95.8% of the isolates were of serotypes/groups included in the current 23-valent polysaccharide vaccine, 52.2% were of serotypes/groups included in the 7-valent conjugated vaccine and 85.5% were of serotypes/groups included in the 11-valent conjugated vaccine.

Adolescent↗

Peptidyl transferase antibiotics perturb the relative positioning of the 3'-terminal adenosine of P/P'-site-bound tRNA and 23S rRNA in the ribosome.

A range of antibiotic inhibitors that act within the peptidyl transferase center of the ribosome were examined for their capacity to perturb the relative positioning of the 3' end of P/P'-site-bound tRNA and the Escherichia coli ribosome. The 3'-terminal adenosines of deacylated tRNA and N-Ac-Phe-tRNA were derivatized at the 2 position with an azido group and the tRNAs were cross-linked to the ribosome on irradiation with ultraviolet light at 365 nm. The cross-links were localized on the rRNA within extended versions of three previously characterized 23S rRNA fragments F1', F2', and F4' at nucleotides C2601/A2602, U2584/U2585 (F1'), U2506 (F2'), and A2062/C2063 (F4'). Each of these nucleotides lies within the peptidyl transferase loop region of the 23S rRNA. Cross-links were also formed with ribosomal proteins L27 (strong) and L33 (weak), as shown earlier. The antibiotics sparsomycin, chloramphenicol, the streptogramins pristinamycin IA and IIA, gougerotin, lincomycin, and spiramycin were tested for their capacity to alter the identities or yields of each of the cross-links. Although no new cross-links were detected, each of the drugs produced major changes in cross-linking yields, mainly decreases, at one or more rRNA sites but, with the exception of chloramphenicol, did not affect cross-linking to the ribosomal proteins. Moreover, the effects were closely similar for both deacylated and N-Ac-Phe-tRNAs, indicating that the drugs selectively perturb the 3' terminus of the tRNA. The strongest decreases in the rRNA cross-links were observed with pristinamycin IIA and chloramphenicol, which correlates with their both producing complex chemical footprints on 23S rRNA within E. coli ribosomes. Furthermore, gougerotin and pristinamycin IA strongly increased the yields of fragments F2' (U2506) and F4' (U2062/C2063), respectively. The results obtained with an RNAse H approach correlate well with primer extension data implying that cross-linking occurs primarily to the bases. Both sets of data are also consistent with the results of earlier rRNA footprinting experiments on antibiotic-ribosome complexes. It is concluded that the antibiotics perturb the relative positioning of the 3' end of the P/P'-site-bound tRNA and the peptidyl transferase loop region of 23S rRNA.

Adenosine↗

Biochemical and kinetic characteristics of the interaction of the antitumor antibiotic sparsomycin with prokaryotic and eukaryotic ribosomes.

Using 125I-labeled phenol-alanine sparsomycin, an analogue of sparsomycin having higher biological activity than the unmodified antibiotic, we studied the requirements and the characteristics of its interaction with the ribosome. The drug does not bind to either isolated ribosomal subunits or reconstituted whole ribosomes. For sparsomycin binding to 70S and 80S ribosomes, the occupation of the peptidyltransferase P-site by an N-blocked aminoacyl-tRNA is a definitive requirement. The sparsomycin analogue binds to bacterial and yeast ribosomes with Ka values of around 10(6) M-1 and 0.6 x 10(6) M-1, respectively, but its affinity is probably affected by the character of the peptidyl-tRNA bound to the P-site. Chloramphenicol, lincomycin, and 16-atom ring macrolides compete with sparsomycin for binding to bacterial ribosomes, but streptogramins and 14-atom ring macrolides do not. Considering the reported low affinity of puromycin for bacterial ribosomes, this antibiotic is also a surprisingly good competitor of sparsomycin binding to these particles. In the case of yeast ribosomes, blasticidin is a relatively good competitor of sparsomycin interaction, but anisomycin, trichodermin, and narciclasin are not. As expected, puromycin is a poor competitor of the binding in this case. The results from competition studies carried out with different sparsomycin analogues reveal, in some cases, a discrepancy between the drug ribosomal affinity and its biological effects. This suggests that some intermediate step, perhaps a ribosomal conformational change, is required for the inhibition to take place.

Binding, Competitive↗

Antibiotic effects on the photoinduced affinity labeling of Escherichia coli ribosomes by puromycin.

The effect of ribosomal antibiotics on the photoinduced affinity labeling of Escherichia coli ribosomes by puromycin [Cooperman, B.S., Jaynes, E.N., Brunswick, D.J., & Luddy, M.A. (1975) Proc. Natl. Acad. Sci. U.S.A. 72, 1974; Jaynes, E.N. Jr., Grant, P.G., Giangrande, G., Wieder, R., & Cooperman, B.S. (1978) Biochemistry 17, 561] has been studied. Although blasticidin S, sparsomycin, lincomycin, and erythromycin are essentially without effect, major changes are seen on addition of either chloramphenicol or tetracycline. The products of photoincorporation have been characterized by one- and two-dimensional gel electrophoresis and by specific immunoprecipitation with antibodies to ribosomal proteins. In the presence of chloramphenicol, protein S14 becomes the major labeled protein. In the presence of tetracycline, L23 remains the major labeled protein, but the yield of labeled ribosomes is enormously increased, and the labeling is more specific for L23. These results are discussed in terms of the known modes of action of these antibiotics and the photoreactivity of tetracycline.

Affinity Labels↗

Pyrrolo[1,4]benzodiazepine antibiotics. Biosynthesis of the antitumor antibiotic 11-demethyltomaymycin and its biologically inactive metabolite oxotomaymycin by Streptomyces achromogenes.

11-Demethyltomaymycin, an antitumor antibiotic produced by Streptomyces achromogenes, and its biologically inactive metabolite oxotomaymycin are biosynthesized from L-tyrosine, DL-tryptophan, and L-methionine. The anthranilate part of 11-demethyltomaymycin is derived from tryptophan probably via the kynurenine pathway. The predominant loss of tritium from DL-[5-3H]tryptophan, during its conversion to 11-demethyltomaymycin and oxotomaymycin is interpreted to mean by NIH shift rules, that the main pathway to the 5-methoxy-4-hydroxy anthranilate moiety is through hydroxylation at C-8 prior to hydroxylation at C-7. The methoxy carbon is derived from the S-methyl group of methionine by transfer of an intact methyl group. The ethylideneproline moiety of 11-demethyltomaymycin is biosynthesized from tyrosine, without a 1-carbon unit from methionine. The results of biosynthetic feeding experiments with L-[1-14C, 3- or 5-3H]tyrosine are consistent with a "meta" or extradiol cleavage of 6,7-dihydroxycyclodopa as has also been demonstrated previously for anthramycin and lincomycin A. An experiment in which L-[1-14C, Ala-2,3-3H]tyrosine was fed showed that both the beta hydrogens of this amino acids are retained in 11-demethyltomaymycin. It has been demonstrated in cultures and washed cell preparations that 11-demethyltomaymycin is enzymatically converted to oxotomaymycin by an intracellular constitutive enzyme. Conversion of oxotomaymycin to 11-demethyltomaymycin by these same preparations could not be demonstrated. The enzymatic activity associated with the conversion of 11-demethyltomaymycin to oxotomaymycin is not limited to the 11-demethyltomaymycin to oxotomaymycin is not limited to the 11-demethyltomaymycin production phase, since trophophase cells and even cells from 11-demethyltomaymycin nonproducing cultures of S. achromogenes were equally active in converting 11-demethyltomaymycin to oxotomaymycin.

5-Hydroxytryptophan↗

UV-B radiation-induced donor- and acceptor-side modifications of photosystem II in the cyanobacterium Synechocystis sp. PCC 6803.

We studied the effect of UV-B radiation (280-320 nm) on the donor- and acceptor-side components of photosystem II in the cyanobacterium Synechocystis sp. PCC 6803 by measuring the relaxation of flash-induced variable chlorophyll fluorescence. UV-B irradiation increases the t(1/2) of the decay components assigned to reoxidation of Q(A)(-) by Q(B) from 220 to 330 micros in centers which have the Q(B) site occupied, and from 3 to 6 ms in centers with the Q(B) site empty. In contrast, the t(1/2) of the slow component arising from recombination of the Q(A)Q(B)(-) state with the S(2) state of the water-oxidizing complex decreases from 13 to 1-2 s. In the presence of DCMU, fluorescence relaxation in nonirradiated cells is dominated by a 0.5-0.6 s component, which reflects Q(A)(-) recombination with the S(2) state. After UV-B irradiation, this is partially replaced by much faster components (t(1/2) approximately 800-900 micros and 8-10 ms) arising from recombination of Q(A)(-) with stabilized intermediate photosystem II donors, P680(+) and Tyr-Z(+). Measurement of fluorescence relaxation in the presence of different concentrations of DCMU revealed a 4-6-fold increase in the half-inhibitory concentration for electron transfer from Q(A) to Q(B). UV-B irradiation in the presence of DCMU reduces Q(A) in the majority (60%) of centers, but does not enhance the extent of UV-B damage beyond the level seen in the absence of DCMU, when Q(A) is mostly oxidized. Illumination with white light during UV-B treatment retards the inactivation of PSII. However, this ameliorating effect is not observed if de novo protein synthesis is blocked by lincomycin. We conclude that in intact cyanobacterium cells UV-B light impairs electron transfer from the Mn cluster of water oxidation to Tyr-Z(+) and P680(+) in the same way that has been observed in isolated systems. The donor-side damage of PSII is accompanied by a modification of the Q(B) site, which affects the binding of plastoquinone and electron transport inhibitors, but is not related to the presence of Q(A)(-). White light, at the intensity applied for culturing the cells, provides protection against UV-B-induced damage by enhancing protein synthesis-dependent repair of PSII.

Cyanobacteria↗

Effects of a complex mixture of therapeutic drugs at environmental levels on human embryonic cells.

The potential risk associated with the presence of low levels of pharmaceuticals in aquatic environments is currently under debate. In this study we investigated the effects of 13 drugs merged to mimic both the association and low concentration (ng/L) profiles detected in the environment. The mixture comprised atenolol, bezafibrate, carbamazepine, cyclophosphamide, ciprofloxacin, furosemide, hydrochlorothiazide, ibuprofen, lincomycin, ofloxacin, ranitidine, salbutamol, and sulfamethoxazole. At environmental exposure levels, the drug mix inhibited the growth of human embryonic cells HEK293, with the highest effect observed as a 30% decrease in cell proliferation compared to controls. Pharmaceuticals activated stress-response signaling protein kinases (ERK1/2), and induced overexpression of glutathione-S-transferase P1 gene. No evidence was found for apoptosis or necrosis in HEK293 cells, although morphological changes were observed. The drug mixture effectively stimulated the expression of cell-cycle progression-mediating genes p16 and p21, with a slight accumulation of cells in the G2/M phase of the cell-cycle. Our results suggest that a mixture of drugs at ng/L levels can inhibit cells proliferation by affecting their physiology and morphology. This also suggests that water-borne pharmaceuticals can be potential effectors on aquatic life.

Cell Cycle↗

Changes in chlorophyll fluorescence during exposure of Dunaliella tertiolecta to UV radiation indicate a dynamic interaction between damage and repair processes.

Photosynthesis in the green alga Dunaliella tertiolecta, as measured by chlorophyll fluorescence, is inhibited by ultraviolet radiation and specifically, under the conditions used, by UVB radiation (UVBR). The decline in the fluorescence parameters F(v)/F(m) and DeltaF/F'(m) under constant UVBR is a first-order function of time of exposure. The data are well-described by the Kok (1956) model, which assumes a dynamic interaction between damage and repair, with repair being proportional to the pool size of inactivated targets. The pattern of photoinhibition is also consistent with the Kok model, in that it shows an initial, approximately linear phase which is time-dependent (reciprocity holds), a transition phase and then an asymptotic phase, representing an equilibrium between damage and repair, which is determined by UVBR fluence rate (reciprocity fails). Photoinhibition in the presence of lincomycin, a protein synthesis inhibitor, is consistent with the cessation of repair processes and, under these conditions, photoinhibition is proportional to exposure time.

Journal Article↗

Suppression of zeaxanthin formation does not reduce photosynthesis and growth of transgenic tobacco under field conditions.

Tobacco (Nicotiana tabacum cv. Xanthi) transformed with an antisense cDNA construct of violaxanthin de-epoxidase (VDE) was examined for the effects of suppressed xanthophyll-cycle activity on photoinhibition, photosynthesis and growth under field conditions. De-epoxidation of violaxanthin and non-photochemical quenching were highly inhibited in antisense plants relative to vector-control and wild-type plants. However, no differences were observed between antisense and control plants in photosynthetic CO(2) uptake and maximum photochemical yield [(F(m)-F(o))/F(m)] measured at predawn or in actual photochemical yield [(F(m)'-F(s))/F(m)'] measured at midday. Moreover, growth rates of the plants were the same, as were the leaf area ratio, plant height and leaf number. Similarly, antisense plants did not exhibit greater susceptibility to photoinhibition than controls under field conditions. In contrast, when chloroplast protein (D1) synthesis was inhibited by lincomycin, antisense plants were more vulnerable to photoinhibition than wild-type plants. These results indicate that photoprotection under field conditions is not strictly dependent on the levels of the de-epoxidized xanthophylls, antheraxanthin and zeaxanthin.

Journal Article↗