Increasing importance of plasmid-mediated trimethoprim resistance in enterobacteria: two six-month clinical surveys.

All clinical isolates of enterobacteria received at the laboratory were monitored for trimethoprim resistance over six months in 1978. The survey was repeated in 1979 and the incidence of trimethoprim resistance showed a slight decrease, but the proportion of resistant strains owing their trimethoprim resistance to transferable R plasmids had almost trebled. There was also a large increase in the proportion of resistant strains exhibiting high-level non-transferable trimethoprim resistance. These findings suggest transposition of genes conferring trimethoprim resistance from plasmids to the bacterial chromosome.     

Trimethoprim resistance in Finland after five years' use of plain trimethoprim.

A total of 1388 urinary bacterial pathogens were tested for resistance to plain trimethoprim after five years'' use of this drug for prophylaxis against urinary tract infections. Samples were obtained in Turku, Finland, where use of the drug is much greater than in other parts of Finland. Resistance to trimethoprim (greater than 8 mg/l; agar-dilution method) occurred in 20.3% of strains isolated from outpatients and 39.8% of strains isolated from inpatients. Escherichia coli and Micrococcus showed low incidences of resistance (11% and 13% respectively in ouptatients and 23% and 19% respectively in inpatients); Enterobacter, Streptococcus faecalis, and Staphylococcus epidermidis occupied an intermediate position; and Proteus mirabilis and Klebsiella were resistant in 41-76% of cases. Similar incidences of resistance were observed to sulphamethoxazole-trimethoprim, sulphamethoxazole, ampicillin, and nitrofurantoin. These findings together with the rare occurrence of side effects and convenient dosage confirm the usefulness of plain trimethoprim for urinary tract infection.     

变性梯度凝胶电泳法研究断奶仔猪粪样细菌区系变化

利用PCR和DGGE技术分析了12头仔猪在断奶后其粪样细菌区系的变化。粪样细菌16S rDNA的V6～V8可变区经PCR扩增,扩增产物经DGGE电泳后再进行相似性分析。结果表明,仔猪断奶当天粪样 DGGE谱带少,同窝仔猪间图谱相似。断奶后,随着断奶时间的推移,每头仔猪的DGGE图谱带逐渐增多,变得复杂和多样,仔猪个体间DGGE图谱差异逐渐增大。仔猪是否同窝以及所采食日粮类型对DGGE图谱没有明显影响。相似性分析还表明,日粮中添加寡果糖的仔猪在断奶后第1周,其粪样微生物区系变化迅速,而后缓慢。     

Study of trimethoprim sensitivity in strains of Yersinia pestis isolated from various natural foci of plague

The effect of exogenic addition of purines and pyrimidines on trimethoprim sensitivity was studied in 134 vaccinal and virulent strains of the plague causative agent isolated at various periods in 12 natural plague foci. 74 strains proved to be sensitive to low concentrations of trimethoprim in the cultivation media irrespective of the presence or absence of thymine or thymidine in them. In this respect the strains differed from many other bacterial species which in the presence of thymidine or thymine were resistant to high concentrations of trimethoprim. 60 natural arginine deficient strains of the plague causative agent from Transcaucasia and Mongolia showed high levels of resistance to trimethoprim on media with thymine or thymidine. The possible mechanism of the plague microbe sensitivity to trimethoprim in the presence of thymine or thymidine is discussed with an account of the literature and original data.     

Genetic analysis of chromosomal resistance to trimethoprim derived from clinical isolates of Escherichia coli

Chromosomal genes conferring resistance to trimethoprim were transferred from three independently isolated thy+ clinical strains of Escherichia coli to Escherichia coli K12 by using P1 transduction. Trimethoprim-resistant transductants were obtained less frequently than transductants of other chromosomal markers, suggesting that there were problems related to the expression of the trimethoprim resistance genes in E. coli K12. Mapping studies revealed that one of the resistance determinants was located at a similar position on the chromosome (1 min) to the fol-type mutations previously described in E. coli K12. The two remaining resistance determinants mapped at separate positions between 2.5 and 3 min on the chromosome. The presence of one of these determinants reduced the efficiency with which either donor or recipient cells carrying it could participate in conjugation mediated by the sex factor F and also resulted in phenotypic interaction with the azi gene. The mechanisms of trimethoprim resistance in the three clinical E. coli isolates studied were more complex and diverse than was expected from previous studies of E. coli K12 mutants.     

Drug resistance in Shigella dysenteriae,S flexneri and S boydii in England and Wales: increasing incidence of resistance to trimethoprim.

A total of 2753 strains of shigella belonging to subgroups A, B, and C that were isolated from patients in England and Wales during the period from 1979 to mid-1983 were studied. Of these, 1690 (61%) were from patients recently returned from abroad or in contact with recent travellers, and 760 (45%) of these affected travellers from the Indian subcontinent. The number of strains resistant to sulphonamides and streptomycin remained at a high level throughout (average 76% and 72% respectively). Resistance to tetracyclines, ampicillin, and chloramphenicol rose, reaching 63%, 51%, and 48%, respectively, in 1982. Strains resistant to trimethoprim were seen in substantial numbers for the first time and increased from 1.3% of all strains in 1979 to 9.9% in 1982 and 16.8% in the first half of 1983. The proportion of patients with recent foreign contact was notably smaller among those with strains resistant to trimethoprim than among those with strains sensitive to trimethoprim. The increase in resistance to trimethoprim might partly result from the use in Britain of compounds containing trimethoprim for the treatment of shigellosis.     

Regulatory changes in the formation of chromosomal dihydrofolate reductase causing resistance to trimethoprim

High resistance to trimethoprim mediated by the several hundredfold overproduction of the drug target enzyme, dihyrofolate reductase, in a clinically isolated Escherichia coli strain, 1810, was cloned onto several vector plasmids and seemed to be comprised of a single dihydrofolate reductase gene, which by DNA-DNA hybridization and restriction enzyme digestion mapping was very similar to the corresponding gene of E. coli K-12. Determination of mRNA formation in the originally isolated resistant strain and strains with cloned trimethoprim resistance determinant demonstrated an about 15-fold increase in production of dihydrofolate reductase mRNA compared with that in E. coli K-12. This was explained by the occurrence of a promoter up mutation in the resistant isolate accompanied by changes in the restriction enzyme digestion pattern found by comparison with the corresponding pattern from E. coli K-12.     

Yeast mutants sensitive to trimethoprim

Wild-type strains of Saccharomyces cerevisiae are resistant to growth inhibition by the folate antagonist trimethoprim. A mutant strain sensitive to trimethoprim was isolated. It was found to be sensitive to both ultraviolet light and X-irradiation. Genetic tests revealed that it was allelic with a known radiation-sensitive strain of Saccharomyces cerevisiae, rad 6-1. Strains harbouring a variety of mutant alleles conferring radiation-sensitivity were tested for sensitivity to trimetroprim. It was found that rad 6-1 and each of the four known alleles of rad 18 conferred sensitivity to the drug, but all other rad mutants tested were trimethoprim-resistant. All trimethoprim-sensitive strains, including double mutants of rad 6 rad 18, gave rise to trimethoprim-resistant outgrowths at a rather high frequency (∼ 10⁻⁵). Several resistant outgrowths were analysed. A wide variation in phenotype with respect to UV-sensitivity was found. Genetical analysis revealed that resistance to trimethoprim resulted from torward mutations at separate loci rather than back mutations of rad 6 or rad 18 alleles.     

A genotyping investigation of the colonization of piglets by Campylobacter coli in the first 10 weeks of life

Aims:  To examine the Campylobacter genotypes colonizing a litter of piglets during the first 10 weeks of life and compare them with those of the sow.
Methods and Results:  Campylobacters were isolated by direct plating of anal swabs. Piglets ( n  = 6) were sampled six times and five isolates per piglet obtained each time. The sow was also sampled but 20 isolates per sampling obtained. Isolates were genotyped by random amplification of polymorphic DNA, pulsed field gel electrophoresis and polymerase chain reaction/restriction fragment length polymorphism of the flagellin gene.
Initially piglets were colonized by Campylobacter coli genotypes from the mother but after 66 days 33% of piglet isolates (based on a mean of the three-genotyping methods) were from other sources. The sow died after 14 days and the initial Campylobacter flora of the foster sow was subsequently replaced by genotypes from the piglets mother. However these constituted only a minor part of her flora after 52 days. Both foster sow and piglets carried multiple genotypes of Camp. coli : up to four in a single piglet sample and seven in one from the sow.
Significance and Impact of the Study:  Piglets are initially colonized by Camp. coli genotypes from their mother but later other genotypes displace them. Colonization is dynamic with the sow able to acquire genotypes from the piglets.
Conclusions:  The large numbers of Camp. coli genotypes carried by pigs, and frequent successions of dominant types, will render epidemiological studies problematic.     

Clinical Evaluation of Co-trimoxazole and Furazolidone in Treatment of Shigellosis in Children

Co-trimoxazole (trimethoprim—sulphamethoxazole) was compared with furazolidone in the treatment of shigellosis in two groups of 33 and 30 patients respectively. Those treated with co-trimoxazole recovered more quickly; none had shigellae in the faeces four days after the start of treatment, whereas in the group given furazolidone eight still had positive stool cultures seven days after treatment.The susceptibility of 104 shigella strains to seven antimicrobial agents was studied by plate dilution technique. All agents but tetracycline and chloramphenicol were found highly effective against most of the strains tested. All shigella isolates were resistant to sulphamethoxazole, and 63% were sensitive to trimethoprim. Potentiation of trimethoprim by sulphamethoxazole was shown in that all strains tested became sensitive to the combination of trimethoprim and sulphamethoxazole in a ratio of 1:20.     

R-factor mediated dihydrofolate reductases which confer trimethoprim resistance.

Six different R-factors conferring trimethoprim resistance had been isolated from a variety of sources. The trimethoprim-resistant dihydrofolate reductases (EC 1.5.1.3) from strains containing these R-factors were purified by ammonium sulphate precipitation and DEAE-cellulose ion-exchange chromatography. The enzymes showed no significant differences in molecular weight, pH profile, substrate profile, heat sensitivity, inhibition profile and Michaelis-Menten kinetics. There was, however, considerable variation in the specific activity of these enzymes in the same bacterial host. When two Escherichia coli trimethoprimsensitive dihydrofolate reductases were examined as controls, considerable differences between their properties and those of the enzymes mediated by R-factors were detected. The data suggest that one trimethoprim resistance gene could be spreading through the bacterial population, possibly situated on a transposon.     

The transmission of campylobacter in piggeries; an epidemiological study

The campylobacter infection of 10 sows and their piglets was monitored. These pigs werekept on two multiplier farms. Rectal faeces samples were taken from the sows shortly beforelittering and at different intervals after littering. Swab samples of rectal content were taken fromsix piglets per sow at different intervals after birth. Nine sows were shown to be infected withcampylobacter before litter and all sows after litter, with an average colony count of 4·1in log N g^–1 of faeces. Half of the piglets became infected withcampylobacter during the first week of life and 85%, after four weeks. Two genetic subtypingmethods (ERIC-PCR and RFLP) were used to study the relationships between campylobacterisolates from sows and piglets. A large diversity of campylobacter subtypes was found.Nevertheless, piglets and their mothers often harboured campylobacter isolates with identicalgenetic subtyping profiles, suggesting that piglets become infected via their mothers. However,observed similarities in genetic subtyping profiles between campylobacters isolated on differentfarms made this difficult to prove.     

Persistence of antibiotic resistance: evaluation of a probiotic approach using antibiotic-sensitive Megasphaera elsdenii strains to prevent colonization of swine by antibiotic-resistant strains

Megasphaera elsdenii is a lactate-fermenting, obligately anaerobic bacterium commonly present in the gastrointestinal tracts of mammals, including humans. Swine M. elsdenii strains were previously shown to have high levels of tetracycline resistance (MIC=64 to >256 μg/ml) and to carry mosaic (recombinant) tetracycline resistance genes. Baby pigs inherit intestinal microbiota from the mother sow. In these investigations we addressed two questions. When do M. elsdenii strains from the sow colonize baby pigs? Can five antibiotic-sensitive M. elsdenii strains administered intragastrically to newborn pigs affect natural colonization of the piglets by antibiotic-resistant (AR) M. elsdenii strains from the mother? M. elsdenii natural colonization of newborn pigs was undetectable (<10⁴ CFU/g [wet weight] of feces) prior to weaning (20 days after birth). After weaning, all pigs became colonized (4 × 10⁵ to 2 × 10⁸ CFU/g feces). In a separate study, 61% (76/125) of M. elsdenii isolates from a gravid sow never exposed to antibiotics were resistant to chlortetracycline, ampicillin, or tylosin. The inoculation of the sow''s offspring with mixtures of M. elsdenii antibiotic-sensitive strains prevented colonization of the offspring by maternal AR strains until at least 11 days postweaning. At 25 and 53 days postweaning, however, AR strains predominated. Antibiotic susceptibility phenotypes and single nucleotide polymorphism (SNP)-based identities of M. elsdenii isolated from sow and offspring were unexpectedly diverse. These results suggest that dosing newborn piglets with M. elsdenii antibiotic-sensitive strains delays but does not prevent colonization by maternal resistant strains. M. elsdenii subspecies diversity offers an explanation for the persistence of resistant strains in the absence of antibiotic selection.     

Thymidine phosphorylase activity and its relationship to trimethoprim in initial strains and thymidine-, thymine-dependent and trimethoprim-resistant mutants of plague microbe]

The comparative study revealed thymidine phosphorylase activity in the initial strains of a plague microbe of the field variety and in thymidine-, thymine-dependent and trimethoprim-resistant mutants of the plague microbe of other varieties. The data fully conformed to the results of the microbiological investigation of the strains' ability to grow on the nutrient media with trimethoprim in the presence of thymine and thymidine. On the basis of these results it appeared possible to divide the initial and mutant strains of the plague microbe into four arbitrary groups: initial strains of the plague microbe of all the varieties except the field ones sensitive to trimethoprim under any temperature conditions of incubation on any medium with any supplements; initial strains of the plague microbe of the field variety resistant to trimethoprim at 28 degrees C in the presence of thymine or thymidine alone; Tmpr mutants whose resistance to trimethoprim at 28 degrees C did not depend on the presence of thymine or thymidine, purine and vitamins, but depended on the presence of these substances at a temperature of 37 degrees C.     

Iclaprim, a novel diaminopyrimidine with potent activity on trimethoprim sensitive and resistant bacteria

Iclaprim, a new selective dihydrofolate inhibitor was synthesized based on rational drug design. Iclaprim's interaction with a resistant Staphylococcus aureus dihydrofolate reductase (DHFR) is outlined in comparison to trimethoprim (TMP). This compound is active against methicillin, TMP and vancomycin resistant strains. Arpida Ltd. is developing Iclaprim for serious hospital infections from Gram-positive pathogens and respiratory tract infections.     

Prevention of drug-resistant Escherichia coli colonization in chickens by treatment with a faecal fluid

This study was performed to prove that intestinal colonization in chickens by resistant Escherichia coli strains present in the environment might be prevented when faeces in which sensitive E. coli strains were dominant was administered to newly hatched chicks. The appearance of resistant E. coli strains was markedly reduced. Escherichia coli O49:H12 was the sensitive E. coli strain which formed the major colonizer in the intestinal tract. In young chickens, this strain persisted as a major component, and even when it was a minor colonizer in the faecal fluid administered, it appeared as a major component soon afterwards. This strain is considered to be a good colonizer in the gut of young chickens.     

Establishment in the piglet gut of lactobacilli capable of degrading mixed-linked beta-glucans

The establishment in piglets of lactobacilli with ability to degrade mixed-linked (1----3), (1----4) beta-D-glucans was studied in faeces from 15 animals. The piglets had free access to creep feed with an estimated content of 2% beta-D-glucans from 5 days of age. On days 3 and 35, ca log 8 cfu/g of beta-glucan-degrading lactobacilli were found in a majority of the samples. On days 7, 14 and 21 such bacteria were only found occasionally. During establishment of the microflora in the neonate, the faecal lactobacilli of the piglet seemed related to those of the sow. Later, the metabolic activity of the lactobacilli in piglet faeces showed a connection to the composition of the diet. The possible relation of these bacteria to occurrence of beta-glucanases attributed to be endogenous in the pig is discussed.     

Prevention of drug-resistant Escherichia coli colonization in chickens by treatment with a faecal fluid

This study was performed to prove that intestinal colonization in chickens by resistant Escherichia coli strains present in the environment might be prevented when faeces in which sensitive E. coli strains were dominant was administered to newly hatched chicks. The appearance of resistant E. coli strains was markedly reduced. Escherichia coli O49:H12 was the sensitive E. coli strain which formed the major colonizer in the intestinal tract. In young chickens, this strain persisted as a major component, and even when it was a minor colonizer in the faecal fluid administered, it appeared as a major component soon afterwards. This strain is considered to be a good colonizer in the gut of young chickens.     

The combination of trimethoprim and sulfamethoxazole in the treatment of urinary tract infection

In a series of 52 patients the combination of trimethoprim and sulfamethoxazole was used for the treatment of urinary tract infection and the results are analyzed with respect to the clinical diagnosis and bacterial etiology. There was complete agreement between in vitro sensitivity and clinical response except in the case of one strain of Streptococcus fecalis. The combination of drugs was effective against some bacterial strains which were resistant to the commonly used antimicrobial agents.