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PubMed2000

[Results of the German investigation in the EU Project "Salmonella in Pork (Salinpork)"--2. Investigations in a slaughterhouse].

In 1997 bacteriological examinations for the distribution of Salmonella in slaughterhouses were carried out in Germany within the framework of an international study "Salmonella in Pork (Salinpork)". During 6 days, 1,200 swab and water samples from slaughtered pigs and the environment were taken. 4.4% of the samples (n = 53) were Salmonella positive. S. typhimurium was isolated mainly (69.8%; n = 37), and 6 phagetypes were differentiated. In addition, S. derby and S. panama could be demonstrated. The resistance pattern of the different isolated S. typhimurium-phagetypes are presented. The phagetype DT 104 was multiresistant to ampicillin, spectinomycin, streptomycin, sulphonamide and tetracycline. In comparison with the serological prevalence of 7.3% of the fattening pigs in the farms (Part 1), only 1.0% of the samples taken from the surface of the carcass were Salmonella-positive. Swabs taken from the liver were in 2.7% positive and samples from the tongue gave in 5.3% of the cases Salmonella-positive results. In the examination of the environment Salmonella was demonstrated mainly from the water outlets, whereas Salmonella could not be isolated from water of the scalding tank. There was only one case (0.7%) in which Salmonella could be isolated from the hands of the personnel, and also only one swab of the polishing machine was positive (1.1%). But 6.7% samples of the saw were Salmonella-positive. A comparison of repeated, at intervals taken samples showed that the number of Salmonella-positive samples was higher in the last examination round of the particular slaughter days. The reason is suspected in the increasing number of slaughtered pigs and supplying farms, which may increase the probability of bringing in Salmonella.

Abattoirs↗
PubMedDate not supplied

[Genetic map and structure in "Escherichia coli" K12 of a resistance plasmid isolated from "Salmonella ordonez" (author's transl)].

A resistance plasmid called R IP173 has been transferred into E. coli K12 from a multiresistant strain of S. ordonez isolated during an epidemic in Dakar. This plasmid mediates for colicine Ib production and resistance to ampicillin, streptomycin, spectinomycin, kanamycin, chloramphenicol, tetracycline and sulfonamides. It is transducible "en bloc" by the phage P1-kc between strains of E. coli K12. Compatibility studies have shown that R IP173 belongs to the fi- class, I1 group. It is transferred "en bloc" in conjugation experiments between E. coli K12 strains. But during transfers from S. ordonez into E. coli, incomplete variants are obtained, lacking different markers. A deletion map was obtained after analysis of 19 different variants, and it is suggested that the loss of markers results from the loss of genetic material during transfer. In this particular case, the deletions observed in transduction or conjugation experiments lead to identical genetic maps.

Ampicillin↗
PubMed1979

[Auxotypes and sensitivity to 6 antibiotics of strains of Neisseria gonorrhoeae isolated at Strasbourg in 1977-1978 (author's transl)].

We have studied 386 strains of Neisseria gonorrhoeae isolated in our laboratory from July 1977 to October 1978, with the Catlin's auxotyping method. The distribution of the four main auxotypes, i. e. (+), (PRO-), (ARG-), (ARG-, HYX-, URA-), is different from those reported by others (in U. S. A. and Sweden). In Strasbourg, the auxotype (+) is prevalent. We have noticed some differences among classes of population. For example (ARG-) and (ARG-, HYX-, URA-) are scarcely isolated from prostitutes and we have found less (ARG-) strains in women than in men. The determination of the minimum inhibitory concentration of 6 antibiotics has shown there was no differences between the auxotypes except for the (ARG-, HYX-, URA-) strains which are more susceptible to all the antibiotics, but spectinomycin. Since August 1978, we have also noticed an increasing of the resistance which is mainly due to (PRO-) strains.

Adult↗
PubMed2001

Gene lmrB of Corynebacterium glutamicum confers efflux-mediated resistance to lincomycin.

The lmrB gene of Corynebacterium glutamicum, which confers specific resistance to lincosamides, such as lincomycin and clindamycin, was isolated. C. glutamicum cells, carrying the lmrB gene in a multicopy plasmid, showed increased resistance to lincomycin with a MIC of 230 microg/ml, which is a 9-fold increase compared to that of the wild type. The lmrB-disrupted mutant became sensitive to the compound. No difference in sensitivity to erythromycin, penicillin G, tetracycline, chloramphenicol, spectinomycin, nalidixic acid, gentamicin, streptomycin, ethidium bromide, and sodium dodecyl sulfate was observed. The protonophore carbonyl cyanide m-chlorophenylhydrazone abolished the lincomycin-resistance of lmrB-carrying cells. The putative protein product of the gene contained 14-transmembrane regions and showed high amino acid-sequence homology to the drug efflux pumps of other organisms. In addition, the putative protein contained a motif for major facilitators, suggesting a role in efflux-mediated resistance to lincomycin.

Amino Acid Sequence↗
PubMedDate not supplied

Antimicrobial susceptibility pattern of Neisseria gonorrhoeae isolated at the Jos University Teaching Hospital, Jos, Central Nigeria.

Widespread, inadequate and inappropriate treatment of gonococcal infections are facilitating antibiotic resistance in Jos Nigeria. We present an antimicrobial susceptibility pattern of Neisseria gonorrhoeae strains in this study. Thirty-one strains of Neisseria gonorrhoeae were isolated by inoculating swabs onto prewarmed heated blood agar and Thayer Martins medium and incubated overnight at 35 to 37 degrees C. The isolates were then screened for antimicrobial susceptibility by the disk diffusion method. About 93.6% were susceptible to azithromycin, 90.3% to ciprofloxacin, 87.1% to ceftriaxone, 80.7% and 64.3% to erythromycin and ofloxacin respectively. Less than half were each susceptible to spectinomycin (38.7%) and gentamicin (32.8%), while 12.9% and 4.6% were susceptible to tetracycline and penicillin respectively. These findings suggest that gentamicin, tetracycline and penicillin may not be recommended in this environment for treating gonococcal infections. Azithromycin, ciprofloxacin and ceftriaxone are the suggested alternatives. Prompt diagnosis, treatment and indeed continued surveillance are hereby recommended for an effective management of gonococcal infections.

Anti-Bacterial Agents↗
PubMed2001

Expression of an antimicrobial peptide via the chloroplast genome to control phytopathogenic bacteria and fungi.

The antimicrobial peptide MSI-99, an analog of magainin 2, was expressed via the chloroplast genome to obtain high levels of expression in transgenic tobacco (Nicotiana tabacum var. Petit Havana) plants. Polymerase chain reaction products and Southern blots confirmed integration of MSI-99 into the chloroplast genome and achievement of homoplasmy, whereas northern blots confirmed transcription. Contrary to previous predictions, accumulation of MSI-99 in transgenic chloroplasts did not affect normal growth and development of the transgenic plants. This may be due to differences in the lipid composition of plastid membranes compared with the membranes of susceptible target microbes. In vitro assays with protein extracts from T(1) and T(2) plants confirmed that MSI-99 was expressed at high levels to provide 88% (T(1)) and 96% (T(2)) inhibition of growth against Pseudomonas syringae pv tabaci, a major plant pathogen. When germinated in the absence of spectinomycin selection, leaf extracts from T(2) generation plants showed 96% inhibition of growth against P. syringae pv tabaci. In addition, leaf extracts from transgenic plants (T(1)) inhibited the growth of pregerminated spores of three fungal species, Aspergillus flavus, Fusarium moniliforme, and Verticillium dahliae, by more than 95% compared with non-transformed control plant extracts. In planta assays with the bacterial pathogen P. syringae pv tabaci resulted in areas of necrosis around the point of inoculation in control leaves, whereas transformed leaves showed no signs of necrosis, demonstrating high-dose release of the peptide at the site of infection by chloroplast lysis. In planta assays with the fungal pathogen, Colletotrichum destructivum, showed necrotic anthracnose lesions in non-transformed control leaves, whereas transformed leaves showed no lesions. Genetically engineering crop plants for disease resistance via the chloroplast genome instead of the nuclear genome is desirable to achieve high levels of expression and to prevent pollen-mediated escape of transgenes.

Adaptation, Physiological↗
PubMed2001

Increased sensitivity to protein synthesis inhibitors in cells lacking tmRNA.

tmRNA (also known as SsrA or 10Sa RNA) is involved in a trans-translation reaction that contributes to the recycling of stalled ribosomes at the 3' end of an mRNA lacking a stop codon or at an internal mRNA cluster of rare codons. Inactivation of the ssrA gene in most bacteria results in viable cells bearing subtle phenotypes, such as temperature-sensitive growth. Herein, we report on the functional characterization of the ssrA gene in the cyanobacterium Synechocystis sp. strain PCC6803. Deletion of the ssrA gene in Synechocystis resulted in viable cells with a growth rate identical to wild-type cells. However, null ssrA cells (deltassrA) were not viable in the presence of the protein synthesis inhibitors chloramphenicol, lincomycin, spiramycin, tylosin, erythromycin, and spectinomycin at low doses that do not significantly affect the growth of wild-type cells. Sensitivity of deltassrA cells similar to wild-type cells was observed with kasugamycin, fusidic acid, thiostrepton, and puromycin. Antibiotics unrelated to protein synthesis, such as ampicillin or rifampicin, had no differential effect on the deltassrA strain. Furthermore, deletion of the ssrA gene is sufficient to impair global protein synthesis when chloramphenicol is added at sublethal concentrations for the wild-type strain. These results indicate that ribosomes stalled by some protein synthesis inhibitors can be recycled by tmRNA. In addition, this suggests that the first elongation cycle with tmRNA, which incorporates a noncoded alanine on the growing peptide chain, may have mechanistic differences with the normal elongation cycles that bypasses the block produced by these specific antibiotics. tmRNA inactivation could be an useful therapeutic target to increase the sensitivity of pathogenic bacteria against antibiotics.

Chloramphenicol↗
PubMed1979

Disseminated gonococcal infections (DGI).

Neisseria gonorrhoeae infects superficial membranes of the eyes, oropharynx, genital tract, and rectum prior to dissemination. Gonococcal isolates cultured from patients with disseminated gonococcal infections (DGI) show resistance to serum bacteriolysis, are very sensitive to penicillin, and have characteristic growth requirements for certain amino acids. DGI is characterized by recurrent chills and fever, polyarthralgias and/or polyarthritis (with effusions), and skin lesions. The skin manifestations of DGI include vesicopustules, hemorrhagic bullae, and petechiae. These lesions are found over the juxta-articular areas of the hands (extensor surfaces) or the feet (dorsal aspects). Focal and disseminated gonococcal infections are now treated with several types of penicillin regimens, tetracycline, or spectinomycin.

Anti-Bacterial Agents↗
PubMed2002

[A novel resolution vector with Bacillus thuringiensis plasmid replicon ori44].

The resolution recognization sites of transposon Tn4430 of Bacillus thuringiensis was inserted into cloning vector pRSET B and pUC19, resulting recombinant plasmids pBMB1201 and pBMB1202. Both of the mini res fragments, BamHI/HindIII fragment in pBMB1201 and EcoRI/HindIII fragment in pBMB1202, were ligated to the 3.3 kb EcoRI/HindIII fragment of shuttle vector pHT3101, which contained the ori. Ec, ampr and emr antibiotic resistant genes, resulting recombinant plasmid pBMB1203. After deleted the BamHI and EcoRI sites which located ouside the two res sites, resolution vector pBMB1204 was resulted. There are multiple cloning sites between two copies of resolution sites which have the same direction. The plasmid replication origin ori44, which come from B. thuringiensis sub sp. kurstaki strain YBT-1520, was inserted into the multiple cloning sites of pBMB1204 and then resolution shuttle vector pBMB1205 was obtained. With spectinomycin resistant gene as target, it was found that the resolution rate is 100% and the stability of the resolved plasmid is 93%. Using this shuttle vector, antibiotic resistance markers and other non-B. thuringiensis DNA can be selectively eliminated after the selection of transformants by antibiotic resistance marker. This vector is very useful to solve the gene safety problem while has no effect on target gene expression.

Bacillus thuringiensis↗
PubMed2001

Antibiogram of Klebsiella pneumoniae isolates from Buea, Cameroon.

OBJECTIVE: To determine the antibiotic susceptibility of K. pneumoniae isolates from Buea, Cameroon. DESIGN: A prospective study of K. pneumoniae isolates from clinical samples of nosocomial origin. SETTING: A laboratory based investigative study at the Biotechnology Centres of the Universities of Buea and Yaounde 1, Cameroon, and three Buea based hospitals. K. pneumoniae isolates were obtained from sputum, wound swabs and urine and screened for their antibiogram using standard procedures. RESULTS: Results on the antibiogram showed seven distinct antibiotypes distinguished by different susceptibilities to aminoglycosides (Spectinomycin and Gentamicin), Chloramphenicol and Augmentin. All the isolates shared multi-resistance to Amoxicillin and Trimethoprim. However, the isolates showed marked susceptibilities to Norfloxacin (90.01%), Cefuroxime (95.45%) and Ciprofloxacin (86.36%). CONCLUSION: The study has revealed that K. pneumoniae isolates in the environment of Buea, Cameroon are multi-drug resistant. This finding is of clinical and epidemiological significance.

Cameroon↗
PubMed1975

Epidemiology of methicillin-resistant Staphylococcus aureus in dairy herds.

Sixty-eight methicillin-resistant Staphylococcus aureus strains were isolated from mastitis milk samples originating from 20 Belgian dairyherds. All these strains appeared to be representatives of one single strain which was probably of human origin. Evidence is presented indicating a rapid in vivo evolutionary change in this strain. The following characteristics were found to be variable: the production of beta haemolysin inversely connected with fibrinolysin (staphylokinase) activity; the production of lipase, enterotoxin B and delta haemolysin; the resistance to neomycin, chloramphenicol, tetracyclines, methicillin and spectinomycin associated with constitutive or inducible macrolide resistance.

Animals↗
PubMedDate not supplied

[Obtaining and analysis of intergeneric somatic hybrids between Brassica napus and "albino" line of Orychophragmus violaceus].

The Orychophragmus violaceus chlorophylldefective line of "albino" type has been obtained by spectinomycin treatment. Somatic hybridization between Orychophragmus violaceus and Brassica napus was performed by fusion of green mesophyll protoplasts of rape and callus protoplasts of the O. violaceus "albino" line. Near two hundred of regenerant plants were selected according to the regeneration type and ability to become green, and were determined as hybrids. Chloroplast DNA in selected hybrids was identical to rape chlDNA, which was confirmed by the PCR-RFLP analysis of plastid DNA fragments. Fragments of hybrid mitochondrial DNA analyzed by the PCR-RFLP analysis were identical to fragments of O. violaceus. The nuclear genome of the majority of hybrids was represented by the O. violaceus genome, which was demonstrated by analyses of isoenzymes, DNA telomeric sequences, ribosomal and satellite DNAs, and the RAPD analysis. The cytogenetic analysis of a number of lines has shown variability in the number of chromosomes in the obtained lines.

Brassica napus↗
PubMedDate not supplied

Virulence factors of salmonella ser. enteritidis strains isolated in Italy from food-borne outbreaks.

Twelve Salmonella ser. Enteritidis strains phage type 4 isolated in Italy from different food-borne outbreaks were characterized for the expression of different virulence traits, for antibiotic resistance, and for plasmid DNA profile. All the twelve S. Enteritidis strains were able to invade and multiply within HeLa cell monolayers, even if at a lower efficiency if compared to an invasive Shigella flexneri strain. The strains were not hemolytic and produced only a moderate-level cytotoxic effect on HeLa cell monolayers. Moreover, all the strains examined produced mannose-sensitive hemagglutination with chicken erythrocytes but were not able to adhere to tissue culture cells. The strains did not produce the hydroxamate-type siderophore aerobactin or the specific ferric-aerobactin receptor. The S. Enteritidis strains were resistant only to spectinomycin, and eleven strains harbored a 38 MDa non-conjugative plasmid, while one strain harbored a 64 MDa conjugative plasmid which carried a colicinogenic activity-encoding locus. The uniformity of antibiotic resistance pattern, of the plasmid DNA content, and of the virulence factors produced indicated that the S. Enteritidis clinical isolates examined are clonally-related.

Journal Article↗
PubMed2003

[Development of a system of plasmid transfer in the strains Streptomyces avermitilis ATCC 31272 and Saccharopolyspora erythraea ATCC 11635 by the method of intergeneric conjugation].

A new method of plasmid DNA transfer from the donor strain Escherichia coli S17-1 to the erythomycin-producing strain Saccharopolyspora erythraea and avermectin-producing strain Streptomyces avermitilis via intergeneric conjugation was proposed. The optimal parameters of the method were chosen for increasing the efficiency of crosses and ensuring easily reproducible results. The behavior of the multicopy plasmid pPM803 and the integration vector pTO1 along with a number of new plasmids specially created by us, was examined in these strains. A new plasmid vector (pSI60) capable of integrating into the chromosome of actinomycetes at the integration site of the temperate actinophage phi C31 was constructed. This vector possesses unique sites convenient for cloning and may be stably maintained in exconjugants of S. avermitilis and in the model strain Streptomyces lividans. The gene encoding resistance to spectinomycin and streptomycin was cloned into the vector pSI60 in this strain. For cloning in strain Sac. erythraea, vectors pSI261-280, which integrate into the chromosome via homology with the cloned DNA and can be stably maintained in exconjugants, were constructed.

Attachment Sites, Microbiological↗
PubMed2003

[Infectious distribution and resistant of Neisseria gonorrhoeae, Mycoplasma, and Chlamydia trachomatis in the chronic prostatitis].

OBJECTIVE: To investigate the infectious distribution and resistant of Neisseria gonorrhoeae, Mycoplasma, and Chlamydia trachomatis in the chronic prostatitis. METHODS: The identification and susceptibility of Neisseria gonorrhoeaes and Mycoplasmas were detected by a cultural method. The nitrocefin test was used to detect the beta-lactamase in Neisseria gonorrhoeae strains. Chlamydia trachomatis was identificated by a monoclonal gold labeled antibody method. RESULTS: A total of 2,900 prostatic fluids were detected and the rates of isolation of Neisseria gonorrhoeae, Mycoplasma, and Chlamydia trachomatis were 3.3%, 12.8%, and 0.9%, respectively. The prevalence of beta-lactamase in Neisseria gonorrhoeae strains was 12.6%. The resistant percentages of Neisseria gonorrhoeae strains were 73.7% to penicillin and 91.6% to ciprofloxacin and ofloxacin respectively, but the susceptibility to spectinomycin, cephalosporin and cefoxitin was good. The resistant rates of Mycoplasma to tetracycline, acetylspiramycin, erythromycin, and ofloxacin were 50.0% or more, but the susceptibilities to roxithromycin, doxycyclin, levofloxacin, minocyclin, josamycin, and azithromycin were about 70.0%-80.0%. CONCLUSION: The isolation rate of Mycoplasma is higher than that of Neisseria gonorrhoeae and Chlamydia trachomatis in the chronic prostatitis. It is important to detect the susceptibility of Neisseria gonorrhoeae and Mycoplasma for the use of antibiotics in reason.

Adult↗
PubMed2003

[Male gonococcal urethritis in the region of Sfax (1996-2000)].

The sexually transmitted infections pose a real public health problem, mainly in developing countries. In Tunisia, the epidemiological state is not precise. The objective of our study was to determine the sociological, epidemiological and bacteriological characteristics of gonococcal urethritis in the area of Sfax. The study was performed on all patients attending at the CNSS of Sfax for urethritis for 5 years, from 1996 to 2000. A urethral swab was carried out for each patient with a classic bacteriological exam. 256 cases were collected. The mean age was 29.7 years. 74.3% were workers. The direct exam was positive in 51.2% of cases and the culture enabled us to isolate 72 strains, of which 15.5% were productive of beta-lactamase and 23.9% were resistant to tetracyclin. There was no resistance to pristinamycin, ofloxacin nor spectinomycin.

Adult↗
PubMedDate not supplied

In vitro susceptibility of avian Escherichia coli and Pasteurella multocida to danofloxacin and five other antimicrobials.

The in vitro susceptibility of Escherichia coli and Pasteurella multocida isolated from poultry was determined to danofloxacin, a novel fluoroquinolone, and five other commonly used antimicrobials. A total of 1737 E. coli field isolates and 107 P. multocida isolates were tested by veterinary diagnostic laboratories in Europe, Japan, South Africa, and North America during the period 1989-91. The antimicrobial susceptibility of these isolates was determined using the Sensititre broth microdilution technique. The minimum inhibitory concentrations (MIC) of danofloxacin, furaltadone, lincomycin, oxytetracycline, spectinomycin, and trimethoprim:sulfamethoxazole that prevented growth of 90% of the bacteria were 0.25 > 64, > 64, > 64, > 128, and > 16 micrograms/ml, respectively, against E. coli isolates and 0.25, 64, 64, 16, 128, and 8 micrograms/ml, respectively, against P. multocida isolates. Danofloxacin demonstrated considerable in vitro potency against these important poultry pathogens, many of which showed extensive resistance to the other antimicrobials tested.

Animals↗
PubMedDate not supplied

[Analysis of nuclear and mitochondrial genomes of transplastomic Salpiglossis sinuata plants obtained by transfer of transformed plastids from N. tabacum (+S. sinuata) cybrid].

New model system of plastid transformation has been proposed using a wild representative of Solanaceae family--S. sinuata. Earlier obtained cybrid plants N. tabacum (+ S. sinuata) were used for transformation experiments by PEG treatment of protoplasts with aadA gene that confers resistance to spectinomycin. Transformed S. sinuata plastome was transferred from N. tabacum (+ S. sinuata) cybrid to S. sinuata wild type plants by somatic hybridization. Molecular analysis of nuclear and mitochondrial DNA has been performed.

DNA, Mitochondrial↗