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Biomedical subjects

A J Hedges

Publications and source records attributed to A J Hedges.

At least 19 recordsLinked to original sources
PubMed2003

Estimating the precision of serial dilutions and colony counts: contribution of laboratory re-calibration of pipettes.

The basic (inherent) precision of serial dilutions and of colony counts made from them may be reliably estimated by reference solely to the pipette-manufacturer's specifications. Such estimates do not include external sources of variation and may be regarded as minima. The quality of estimation can be improved by using information gained by laboratory ('in-house') re-calibration of the pipettes. The degree of improvement was assessed by comparison with similar series made without re-calibration. It was found that improvement was minimal for colony counts but worthwhile for homogeneous solutions.

Bacteria↗
PubMed2002

Estimating the precision of serial dilutions and viable bacterial counts.

The propagation of error in serial dilutions was investigated theoretically and by means of computer simulations. The principal aim of the study was, given only the pipette manufacturer's specification, to estimate the variance of any step in a dilution series both of pure solutions and of homogeneous bacterial suspensions by means of simple formulae. The study was extended to include bacterial plate counts by both the standard and the Miles and Misra methods. It was found that such estimation was possible and that the distributions approximated the normal sufficiently for the construction of confidence intervals (Cls) by the usual method. Such intervals can be regarded as minima which could be inflated by other, possibly undetermined, factors. It is suggested that laboratories could construct tables such as that reported here for pipettes and methods in common use to facilitate estimation. While replication of the final sampling step of a plate count increases the precision of estimation, averaging across dilutions may decrease precision and is not recommended for the standard pour-plate count.

Bacteria↗
PubMed2001

Antimicrobial activity of fluoroquinolone photodegradation products determined by parallel-line bioassay and high performance liquid chromatography.

The fluoroquinolones produce multiple photodegradation products. Little is known about these products, particularly whether any possess antimicrobial activity. To investigate this, we used the parallel-line bioassay to investigate discrepancies in zone of inhibition size in conjunction with high performance liquid chromatography (HPLC) analysis. A continuous flow photochemical reaction unit ('Beam-Boost') was used to partially photodegrade the fluoroquinolones ofloxacin, levofloxacin, ciprofloxacin and moxifloxacin (0.02 mM) by between 15 and 89%, as confirmed by HPLC. The concentration of residual parent fluoroquinolone in each irradiated sample was measured by HPLC and a non-irradiated control solution was prepared at the same concentration. These were compared by parallel-line bioassays using Escherichia coli, Enterobacter cloacae and Klebsiella oxytoca. With ofloxacin and levofloxacin, the zone size for the control solution was significantly less than that of the irradiated solutions, with >15% photodegradation in at least two of the indicator organisms, indicating that the photodegradation products possess antimicrobial activity. No difference was seen with ciprofloxacin at any level of photodegradation with any of the indicator organisms, nor with moxifloxacin at 30 and 54% photodegradation. A significant difference was observed with E. cloacae only, at 83% photodegradation.

Anti-Infective Agents↗
PubMed1997

The pharmacokinetics of intravenous ciprofloxacin 400 mg 12 hourly in patients with severe sepsis: the effect of renal function and intra-abdominal disease.

Serum concentrations of ciprofloxacin were reviewed in 22 patients given ciprofloxacin 400 mg intravenously 12 hourly for severe infection. No dosage modifications were made in patients with renal impairment. Patients who had either bowel or liver pathology in addition to renal failure had significantly higher serum concentrations than all other patients. Dosage reduction of ciprofloxacin in patients with severe sepsis and impaired renal function is not required unless they have co-existent intra-abdominal disease.

Abdomen↗
PubMed1996

A new time-kill method of assessing the relative efficacy of antimicrobial agents alone and in combination developed using a representative beta-lactam, aminoglycoside and fluoroquinolone.

A time-kill curve employing nine sampling times over 6 h was used to provide data which were then used to develop a theoretical (best-fit) curve. From the theoretical curve parameters describing the rate of kill (alpha), time from addition of antibiotic to initiation of killing (d) and a function of the degree of killing observed (Ym/Yo) were defined. The area-under-the-curve (AUC) was calculated from the theoretical curve. The variability of each parameter was assessed using a theoretical curve to fit the data from experiments done on three occasions and in triplicate. In terms of the parameters alpha, d, Ym/Yo and AUC, no synergy was demonstrated with combinations of piperacillin/tazobactam plus ciprofloxacin or gentamicin when compared with single antibiotics. The AUC represents the best summary parameter of a time-kill curve but should be supported by other parameters describing the best-fit curve.

Acinetobacter↗
PubMed1995

A critical assessment of the agar dilution chequerboard technique for studying in-vitro antimicrobial interactions using a representative beta-lactam, aminoglycoside and fluoroquinolone.

The agar dilution chequerboard technique of studying antimicrobial interactions was assessed by testing a representative beta-lactam (piperacillin/tazobactam), aminoglycoside (gentamicin) and fluoroquinolone (ciprofloxacin) against themselves, that is piperacillin/tazobactam plus piperacillin/tazobactam, gentamicin plus gentamicin and ciprofloxacin plus ciprofloxacin. In addition, combinations of piperacillin/tazobactam plus gentamicin or ciprofloxacin were also tested against Enterobacteriaceae and Acinetobacter spp. in triplicate. The agar dilution chequerboard technique did not reliably show addition when agents were combined with themselves, and there was also considerable variation when beta-lactam plus aminoglycoside or fluoroquinolone combinations when tested in triplicate. These observations, and problems with the design of the method, indicate that the chequerboard technique should be used only with adequate controls and replication, and then interpreted with extreme caution; ideally, it should not be used as a method of assessing antimicrobial interactions.

Acinetobacter↗
PubMed1994

An antimicrobial effect associated with rabbit primary aqueous humour.

Viability curves were recorded for a variety of bacteria suspended in rabbit primary aqueous humour. Controls included neat rabbit serum, dilute rabbit serum (1/50), dilute heat treated fetal bovine serum (1/50), and balanced salt solution. Rabbit aqueous humour was bactericidal in vitro for Staphylococcus aureus and Pseudomonas aeruginosa. Micrococcus spp, Streptococcus pneumoniae and Escherichia coli were not affected. The factor mediating the bactericidal effect survives filtration (0.2 micron porosity) and freezing in liquid nitrogen (-196 degrees C) but is labile when stored in air at temperatures of 4 degrees C, 25 degrees C, and 37 degrees C for 1 hour. The bactericidal effect was stable when aqueous humour was stored in an atmosphere of 5% CO2 at 37 degrees C for 3 hours. The antibacterial component in rabbit aqueous humour has yet to be identified.

Animals↗
PubMed1988

Monitoring for the development of antimicrobial resistance during the use of olaquindox as a feed additive on commercial pig farms.

Since 1982, when olaquindox was introduced as a pig-feed additive in the UK, about 12 commercial farms in Suffolk have been monitored annually to check for the possible emergence of resistance to olaquindox and chloramphenicol among the coliform flora of the pigs and their environment. In spite of the sampling variability and the impossibility of controlling the use of feed additives and management on the farms, the overall results obtained were consistent and, it is suggested, the method is widely applicable. A steady, albeit low, increasing incidence and level of resistance to olaquindox was recorded (1982-1984) on farms using it and, to a lesser degree, on neighbouring farms that did not. No significant increase in the level of chloramphenicol resistance was observed. Genetical studies on a selection of olaquindox-resistant isolates suggested that the genes determining resistance were likely to be borne on the chromosome.

Animal Feed↗
PubMed1988

Olaquindox resistance in the coliform flora of pigs and their environment: an ecological study.

Faecal samples were taken weekly from a chosen pen on each of four commercial pig farms, two of which used olaquindox as a feed additive. Coliform bacteria were isolated from the samples and the incidence and level of resistance to olaquindox and to four therapeutic antibiotics determined. The coliforms isolated were biotyped to follow the emergence of drug-resistant sub-populations. The results showed that the coliform flora was complex and that a turnover of biotypes was associated with changes in the occupancy of the pen and, possibly, diet. This turnover led to large fluctuations in both incidence and level of resistance to olaquindox and the antibiotics. Olaquindox resistance was not specifically linked with resistance to any therapeutic antibiotic, and the possible origin of olaquindox-resistant strains is discussed in relation to the biotypes.

Ampicillin↗
PubMed1985

The ecology of Escherichia coli in market calves fed a milk-substitute diet.

Dynamic changes in the Escherichia coli population in the calf gut were studied over 21 days in a group of 18 intensively-reared market calves. Isolates were identified by O-serogrouping, biotyping and resistogram patterns. Seventy O-serogroups were identified among nearly 3000 E. coli isolates examined and these were subdivided into 416 strains by means of their biotype and resistogram. Seventy-five per cent of these strains were detected only once or twice, which points to the continual replacement of the E. coli flora with strains that showed low persistence in the gut. The rise in the frequency of antibiotic resistance observed during the study was not due to a change in the proportion of resistant to sensitive strains in the gut flora. It was a consequence of the displacement of the original flora by multiply-resistant strains, which presumably originated from the calves' environment.

Animal Feed↗
PubMed1985

The ecology of Escherichia coli in calves reared as dairy-cow replacements.

A continual turn-over in the strains forming the majority of faecal Escherichia coli flora was demonstrated in 16 calves reared as dairy cow replacements. The incidence of antibiotic resistance among isolates, as measured by an Antibiotic Resistance Index (ARI), changed markedly with the age of the calf. The value was low initially, when the calves were 1-2 days old and housed with adult animals. It then rose rapidly during the first week after the animals had been weaned and moved into nursery pens. This change in ARI was associated with the isolation of strains resistant to four or five of the six drugs included in the sensitivity test. The ARI then fell from the third week to low levels by the time that the calves were five months of age. This fall was due to the isolation of an increasing proportion of sensitive E. coli strains. These differed from the sensitive strains which had colonized the calves in the early days of life so demonstrating that the change was not due to the reemergence of strains identified several weeks previously. The source of E. coli strains was presumed to be the calfs' environment but further investigations are required to prove this conclusively.

Animals↗
PubMed1975

Distribution of levels of penicillin resistance among freshly isolated strains of N. gonorrhoeae. Application of a novel sensitivity assay.

A novel diffusion zone method of quantitative assay of the antibiotic sensitivity of bacterial strains was tested on freshly isolated gonococci. Smoothly variable estimates of the minimum inhibitory concentration of penicillin for these strains was obtained with sufficient accuracy and precision (coefficient of variation c. 10 per cent.) by means of a simple graphical analysis and without replication. Such estimates were free from the chief sources of error associated with the commonly applied 'incorporation' and 'diffusion' methods. The method revealed that 816 isolates of gonococci obtained in the Bristol area during a 6-month period fell into a large 'sensitive' group (MIC c. 0.02 unit per ml.) and three smaller more resistant groups, and that this pattern occurred in three widely spaced centres within the area. It is suggested that the method is capable of revealing details of distribution that may be masked by the usual techniques and that it is of wide applicability.

England↗
PubMed1973

Reversibility of the specific adsorption of colicin E2-P9 to cells of colicin-sensitive strains of Escherichia coli.

The adsorption of colicin E2-P9 to its specific receptors on cells of sensitive strains of Escherichia coli is reversible under normal experimental conditions. At temperatures above 20 C, colicin may desorb from one cell and be readsorbed by a second with potentially lethal consequences. However, desorption of colicin seems unable to rescue a cell once it has received a lethal dose. These findings have implications both for the nature and types of specific receptors, and for the assay of colicin by the survivor count (lethal unit) methods.

Adsorption↗