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Biomedical subjects

R J Gilbert

Publications and source records attributed to R J Gilbert.

At least 73 records · Page 4Linked to original sources

Synthetic DNA probes for detection of enterotoxigenic Clostridium perfringens strains isolated from outbreaks of food poisoning.

Four synthetic oligonucleotides encoding different parts of the Clostridium perfringens enterotoxin gene were used to test the enterotoxigenicity of C. perfringens strains isolated from confirmed outbreaks of food poisoning. Of the 245 strains isolated from food and feces originating from 186 separate outbreaks, 145 (59%) gave hybridization reactions with each of the four DNA probes used, while 104 strains did not hybridize with any of the probes. There was no correlation between serotype and the presence of the enterotoxin gene, although the C. perfringens enterotoxin gene was rarely detected among nontypable strains (17%). Results show that DNA hybridization is a suitable method for the identification of C. perfringens strains which have the potential to produce enterotoxin.

Base Sequence↗

The survival of salmonellas in shell eggs cooked under simulated domestic conditions.

Strains of Salmonella enteritidis, S. typhimurium and S. senftenberg inoculated into the yolks of shell eggs were found to survive forms of cooking where some of the yolk remained liquid. Survival was largely independent of the size of the initial inoculum. The organisms also grew rapidly in eggs stored at room temperature and after 2 days the number of cells per gram of yolk exceeded log10 8.0. With this level of contamination viable cells could be recovered from eggs cooked in any manner.

Animals↗

A national outbreak of Salmonella typhimurium DT 124 caused by contaminated salami sticks.

An outbreak of Salmonella typhimurium DT 124 infection which affected 101 people in England in December 1987 and January 1988 was detected through surveillance of laboratory reports from medical microbiology laboratories of the NHS and PHLS. Within 1 week of noting the increase in reports, epidemiological and microbiological investigations identified a small German salami stick as the vehicle of infection and the product was withdrawn from sale. The epidemiological investigation highlighted the occurrence of a long incubation period, bloody diarrhoea. Prompt recognition and investigation of the outbreak prevented further cases of severe infection.

Adolescent↗

Infantile botulism.

A 4 month old boy presented with respiratory difficulty and hypotonia. Clostridium botulinum and its toxin were isolated from his faeces and he had electromyographic changes typical of infantile botulism. This is only the second case in the United Kingdom: unfamiliarity with the presentation could result in misdiagnosis.

Botulism↗

Effect of purified Clostridium difficile toxins on intestinal smooth muscle. I. Toxin A.

In these studies we determined the effects of purified Clostridium difficile toxin A, an enterotoxin, on the electrophysiological and contractile properties of rabbit intestinal circular smooth muscle and correlated these effects with changes of smooth muscle morphology. Simultaneous measurements of intracellular membrane potential and contractility were determined in excised ileal muscle strips after administration of toxin A in vivo (60 micrograms/ml) into an isolated rabbit ileal loop or directly in vitro (0.1-60 micrograms/ml) to a normal muscle strip. Toxin A injection in vivo resulted in membrane depolarization and increased slow wave and action potential frequency. Toxin A injection in vivo also caused increased amplitude of spontaneous and carbachol-induced phasic contractions. The electrophysiological effects of in vivo administration of toxin A were correlated with an inflammatory infiltrate of the lamina propria, but no light or electron microscopic evidence of injury to smooth muscle cells was seen. In contrast to the in vivo studies, direct in vitro exposure of normal ileal muscle strips to toxin A had no effect on either spontaneous or carbachol-induced electromechanical activity. Our results indicate that in vivo administration of C. difficile toxin A into a rabbit ileal loop, but not direct in vitro exposure, causes significant alterations of smooth muscle excitation-contraction coupling that may be mediated by products of local inflammatory cells.

Action Potentials↗

Effect of purified Clostridium difficile toxins on intestinal smooth muscle. II. Toxin B.

In the companion paper [Am. J. Physiol. 256 (Gastrointest. Liver Physiol. 19): G759-G766, 1989] we showed that highly purified Clostridium difficile toxin A had a profound effect on intestinal smooth muscle after in vivo but not in vitro exposure. In this study we assessed the effects of in vivo and in vitro exposure to C. difficile toxin B on simultaneous measurements of intracellular membrane potential and contractility in rabbit ileal smooth muscle. Direct exposure of ileal smooth muscle to toxin B (0.1-60 micrograms/ml) in vitro caused membrane depolarization and inhibition of action potential frequency, amplitude, and peak voltage, but no effect on slow wave frequency or amplitude was seen. Toxin exposure also resulted in inhibition of the amplitude of carbachol-induced contractions, with phasic contractions being significantly more sensitive to the effect of toxin B than tonic contractions over the complete dose range. The electromechanical effects of toxin B were not affected by prior administration of tetrodotoxin, atropine, hexamethonium, or phentolamine. In contrast, toxin B administered in vivo into an isolated ileal loop had no effect on spontaneous electromechanical properties of excised smooth muscle strips. Our results indicate that direct exposure in vitro of ileal smooth muscle to C. difficile toxin B causes membrane depolarization in association with inhibition of electromechanical activity. This effect, in combination with the indirect effects of toxin A, may contribute to altered intestinal motility during diarrhea caused by C. difficile.

Action Potentials↗

A comparison of two procedures for the isolation of Listeria monocytogenes from raw chickens and soft cheese.

A cold enrichment method and a modified FDA procedure were compared for the isolation of Listeria monocytogenes from raw chickens and soft cheese. L. monocytogenes was isolated from a total of 23 of 222 cheese and 70 of 160 chicken samples by either one or both methods. Neither method alone yielded all isolates from the two food types. Only 12 cheese and 13 chicken samples were shown to be positive by both methods, although the serotypes isolated were not always identical. On some occasions one method yielded L. monocytogenes while the other produced a different Listeria sp. Reasons for differences in the performance of the two procedures and various points of technical interest are discussed.

Animals↗

The occurrence in the U.K. of Listeria species in raw chickens and soft cheeses.

Retail samples of 100 raw chickens and 222 U.K. and imported soft cheeses were examined for the presence of Listeria species. 60% of raw chickens (fresh and frozen) were contaminated with L. monocytogenes and 28% with other Listeria spp. including L. welshimeri, L. seeligeri and L. innocua. Six serotypes of L. monocytogenes were represented (1/2, 3a, 3b, 3c, 4b, 4d) of which more than one were isolated from some samples. 10% of the soft cheeses examined were found to contain L. monocytogenes at levels from less than 10(2) cfu/g to 10(5) cfu/g. The incidences in cheeses from various countries were Italy (16%), France (14%), Cyprus (10%) and the U.K. (4%). Only 2 serotypes (1/2 and 4b) were isolated, some samples containing both. L. innocua was the only other Listeria sp. found. There was no correlation between either the contamination with E. coli or the processing of the original milk used to make the cheeses (raw or pasteurized) and the presence of L. monocytogenes or other Listeria spp. The contribution of contaminated food to the epidemiology of listeriosis in the U.K. is discussed.

Animals↗

The oral disposition of zinc following the use of an anticalculus toothpaste containing 0.5% zinc citrate.

Zinc is retained in the mouth after use of a toothpaste containing 0.5% zinc citrate. More than one third of the dose was found to be retained after normal brushing. Elevated zinc levels were also found in plaque. Saliva zinc levels were significantly above background for at least 2 h after brushing. In-vitro experiments demonstrated that zinc can bind to the pellicle-coated tooth surface and can subsequently adsorb into saliva. Plaque can calcify to form calculus containing appreciable levels of hydroxyapatite. Zinc adsorbs to hydroxyapatite inhibiting crystal growth. Levels of zinc in plaque were found to be considerably higher than those taken up by hydroxyapatite in an in-vitro test of crystal growth inhibition indicating the potential of zinc to inhibit calculus formation.

Adsorption↗

Plasmid, serotypic, and enterotoxin analysis of Bacillus cereus in an outbreak setting.

Bacillus cereus is a recognized agent of food-borne disease. In this report we describe an outbreak of B. cereus gastroenteritis associated with consumption of beef stew among patients and staff at a Rhode Island nursing home. The beef had been improperly stored after preparation. The predominant symptoms of the illness were cramps and diarrhea; it lasted an average of 16 h. No deaths occurred. The organism was recovered from 10 of 23 stools collected from ill patients and 1 of 21 stools collected from controls (P = 0.0044, Fisher's two-tailed exact test). All isolates had the same biotype and serotype, newly designated H.26; all elaborated the diarrheal B. cereus enterotoxin when tested in rabbits by the vascular permeability reaction; and all had identical plasmid profiles, which differed from those of B. cereus strains selected randomly from other outbreaks. Plasmid analysis may prove to be a useful new tool in investigating outbreaks of B. cereus food poisoning.

Adult↗

Evaluation of ELISA, RPLA, and Vero cell assays for detecting Clostridium perfringens enterotoxin in faecal specimens.

Three hundred and ninety two faecal specimens from 70 separate outbreaks of suspected Clostridium perfringens food poisoning were examined by enzyme linked immunosorbent assay (ELISA), reversed passive latex agglutination (RPLA), and Vero cell assays for the presence of enterotoxin. Although the most time consuming method, ELISA was the most specific and reproducible. RPLA was slightly more sensitive than ELISA, but it showed some non-specific reactions. The Vero cell assay was the least sensitive and least reproducible method, being affected by some non-specific cytotoxic and cytotonic reactions. Normal rabbit serum should be included in the Vero cell assay as a control for the neutralisation of cytotoxic effects.

Animals↗

Salmonella ealing infections associated with consumption of infant dried milk.

In an outbreak of Salmonella ealing infections in November and December, 1985, the proportion of infants infected was in excess of the normal distribution pattern and the cases were geographically widespread. All the infected infants had been fed with a dried-milk product from one manufacturer. Despite intensive efforts to isolate S ealing from packets of the product, it was found only in low numbers in 4 of 267 sealed packets. The source of infection was traced to the factory spray-drier, which had a hole in its inner lining, allowing escape of powder and its return from contaminated insulation material. The plant was closed and the equipment scrapped and later replaced. It is recommended that raw milk and whey, which frequently contain salmonellae, should not be allowed onto the site of milk-drying plants.

Adult↗