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Biomedical subjects

R W Johnston

Publications and source records attributed to R W Johnston.

At least 19 recordsLinked to original sources
PubMed1990

The isolation of salmonellae from poultry environmental samples by several enrichment procedures using plating media with and without novobiocin.

A two-part study was conducted to examine the efficacy of several enrichment-broth techniques and of plating media for detecting salmonellae from poultry environmental samples. The data are reported on pooled samples collected from five poultry houses. The samples were cultured for salmonellae, using up to four different enrichment procedures and employing plating media with and without novobiocin. The primary enrichment-broth procedures were: 1) buffered peptone water preenrichment to Hajna's tetrathionate (TT) broth; and 2) direct inoculation in TT broth. The delayed secondary-enrichment procedure involved prolonged incubation at room temperature and transfer of the primary broths. The plating media consisted of: 1) xylose lysine desoxycholate agar (XLD); 2) xylose lysine desoxycholate agar containing 15 or 20 micrograms per mL of novobiocin (XLDN); 3) brilliant green sulfapyridine agar (BGSP); and 4) brillant green agar containing 20 micrograms per mL of novobiocin (BGN). Of the 94 Salmonella-positive recoveries from the enrichment broths in which complete comparisons could be made, an average of 75% were recovered from the primary enrichment broths and an average of 86% were recovered from the delayed secondary-enrichment broths. Of the 254 Salmonella-positive isolations in which complete comparisons could be made, an average of 65% were isolated on the plating media without novobiocin and an average of 97% were isolated on the plating media containing novobiocin. Overall, the delayed secondary enrichment and enteric plates supplemented with novobiocin significantly improved Salmonella detection from the farm environmental samples.

Animals↗
PubMed1984

Detection and quantitation of chloramphenicol by competitive enzyme-linked immunoassay.

A competitive enzyme-linked immunoassay for the detection and quantitation of chloramphenicol has been developed. The binding of specific rabbit antibody to solid-phase-bound chloramphenicol was competitively inhibited by free chloramphenicol in the sample to be assayed. Antibody not displaced was indicated by using a commercially available, enzyme-linked, anti-rabbit immunoglobulin preparation and reacted with added substrate. Enzyme activity, measured spectrophotometrically, was inversely proportional to the concentration of chloramphenicol in the sample. Quantitation of the antibiotic was linear to 100 ng/ml, with a lower limit of detection of 1 ng/ml (P less than 0.05). Specificity was demonstrated by the lack of inhibition by any of 31 selected antimicrobial agents or chemicals tested in the assay. Chloramphenicol sodium succinate and thiamphenicol, an experimental antibiotic similar in structure to chloramphenicol, were the only drugs found to produce cross-reactions. In addition to excellent sensitivity and specificity, the assay was shown to have good precision and economy and could be completed in approximately 24h.

Animals↗
PubMed1984

Effects of the evisceration and cooling processes on the incidence of Salmonella in fresh dressed turkeys grown under Salmonella-controlled and uncontrolled environments.

In July and December 1981, turkeys grown in a Salmonella-free environment were sampled at the end of the slaughter and evisceration processes to determine the effect of slaughter and evisceration on the incidence of Salmonella in raw, eviscerated turkeys. A total of 109 eviscerated turkeys from the Salmonella-free flocks and 79 control turkeys were sampled. A total of 75 environmental swab samples and 5 chiller water samples were collected. Less than 1% of the eviscerated Salmonella-free turkeys were Salmonella positive and 6.3% of the eviscerated control turkeys were Salmonella positive. The results of the swab samples indicated generally good processing equipment sanitation.

Abattoirs↗
PubMed1983

The microbiology of raw, eviscerated chickens: a ten year comparison.

In 1979 a survey of selected chicken eviscerating plants was conducted to determine the levels of coliforms, Escherichia coli, and Salmonellae sp. on eviscerated chickens under current manufacturing practices. A comparison was made of the data from this survey and one conducted in 1969. The 1979 survey found that the chickens did not have a statistically significant reduced incidence of salmonella compared to chickens analyzed in 1969. Also, much of the bacterial contamination on a carcass at the end of the evisceration line was transient and was readily removed by the final spray washer. The addition of chlorine to chiller water did not appear to have an unusually beneficial effect on the microbiological quality of the chickens. In 1969, in the nine plants studied, the incidence of salmonella on chickens at the exit of the chillers was 20.5%. In 1979, in the nine plants studied, the incidence of salmonella on chickens at the exit of the chillers was 11.6%. In 1979 there was about a 45% reduction in the incidence of salmonella in eviscerated chickens in the nine plants studied compared to the incidence of salmonella in eviscerated chickens in 1969.

Abattoirs↗
PubMed1982

The incidence of Salmonella species and serotypes in young whole chicken carcasses in 1979 as compared with 1967.

A survey was conducted to determine the incidence of salmonellae in processed, ready-to-market, whole young chickens. Carcasses from 15 federally inspected chickens eviscerating plants were analyzed using a carcass washing technique for determining the presence of salmonellae. The results obtained during the 1979 incidence survey were compared to results obtained in an identical 1967 Salmonella survey. In the 1967 study, salmonellae were isolated from 171 ot the 597 (28.6%) whole chickens tetrathionate broth rinsings analysed. In the 1979 study, 222 of 601 (36.9%) of similarly analysed chicken samples were positive. Percentile positive findings from individual plants range from 7.5 to 73.7% in 1967 and from 2.5 to 87.5% in 1979.

Animals↗
PubMed1982

Incidence of Salmonella in fresh dressed turkeys raised under Salmonella-controlled and uncontrolled environments.

The incidence of salmonella in turkeys from experimental salmonella-controlled and uncontrolled, or normal, flocks processed at three turkey slaughter plants were compared. The results indicate that processing salmonella-controlled turkeys in a plant that routinely kills normal birds may result in the contamination of the salmonella-controlled birds, probably due to salmonella in the plant environment. The salmonella-controlled turkeys studied tended to have a lower incidence of salmonella than normal birds. These observation indicate that salmonella control practices in turkey raising can result in a salmonella reduction in market birds even under existing commercial slaughter, evisceration, and cooling procedures.

Abattoirs↗
PubMed1981

Evaluation of iron dextran and mucin for enhancement of the virulence of Yersinia enterocolitica serotype O:3 in mice.

The pathogenic role of Yersinia enterocolitica serotypes O:3, O:8, and O:9 in human infections is well documented. Whereas the virulence of the O:8 strains can be readily demonstrated in mice by 50% lethal dose determinations, the O:3 and O:9 strains have no lethal effect on mice by any route of inoculation. A mouse virulence test for the O:3 and O:9 strains is described. Y. enterocolitica strains were first tested for the presence of virulence-associated plasmid characteristics by auto-agglutination and gel electrophoresis procedures before mouse virulence determinations. The 50% lethal dose of the O:3 strains injected intraperitoneally with 2.5% mucin was about 10(7) colony-forming units. However, histological examinations showed that mucin allowed the growth of Y enterocolitica on the surface of the livers and spleens of the mice without internal lesions. The 50% lethal dose of the same O:3 strains injected intraperitoneally with 1 ml of 10% iron dextran in saline was about 10(5) to 10(6) colony-forming units, and the nonlethal infective dose with typical lesion development was 20 to 200 colony-forming units. The infected mice developed symptoms and extensive liver and spleen lesions which differed from those in mice infected intraperitoneally with the virulent O:8 strains. These results showed that the virulence of the O:3 Y. enterocolitica strains can be measured by intraperitoneal injection with iron dextran. This procedure was used to test the virulence of food isolates, plasmidless strains, and the effect of growth temperatures.

Agglutination↗
PubMed1980

Two modified selenite media for the recovery of Yersinia enterocolitica from meats.

Yersinia enterocolitica is one of the few human pathogens that grows at the proper food refrigeration temperatures of 0 to 5 degrees C. Although the isolation of environmental biotypes of Y. enterocolitica from many types of food and water has been reported in the literature, the recovery of the sensitive strains inoculated into foods has been slow and uncertain. Rapid recovery of several clinical strains inoculated into meats was accomplished by using two modified selenite broths without added nutrients. It was critical to restrict the sample size of the blended meat suspension at the 0.2-g/100 ml level, thereby restricting the growth of the total bacterial population in the selenite enrichment media. Otherwise, the slower growing Y. enterocolitica would be overwhelmed by the faster growing normal bacterial flora from the meat. Both the resistant serotype O:3 and the sensitive O:8 clinical isolates of Y. enterocolitica were recovered from the modified selenite enrichment media after 2 and 3 days of incubation at 22 degrees C.

Alkalies↗
PubMed1980

Rapid detection of staphylococcal thermonuclease on casings of naturally contaminated fermented sausages.

Staphylococcal food poisoning associated with fermented sausages has been a recurring problem. By testing for thermonuclease by direct application of sausage casing disks on the surface of thermonuclease assay agar plates, possible Staphylococcus aureus growth in fermented sausages could be detected simply and rapidly. Koupal-Deibel deoxyribonucleic acid agar was somewhat superior to toluidine blue deoxyribonucleic acid agar for thermonuclease assay of fermented sausage casings. The sensitivity of the thermonuclease casing test was comparable to that of the extraction procedure, and the thermonuclease casing test results were in complete agreement with the thermonuclease assay results by the extraction procedure. The thermonuclease casing test offers government and industry laboratories a useful screening tool which could significantly reduce the problem of staphylococcal enterotoxins in fermented sausages.

Fermentation↗
PubMed1979

Associative properties of butyryl-coenzyme A synthetase from ox liver mitochondria.

Butyryl-coenzyme A synthetase (butyrate:CoA ligase (AMP-forming), EC 6.2.1.2) from an acetone-dried powder of ox liver mitochondria was found to have a molecular weight of approx. 40 000. The sedimentation equilibrium analysis suggested the presence in solution of higher molecular weight forms of the enzyme and these could also be obtained by extracting the enzyme from the mitochondrial powder in non-reducing conditions. The enzyme was inhibited by sulphydryl reagents, and was found to have at least one available thiol group/molecule. The relationship between enzymic activity and concentration was non-linear, and suggested that an inactive monomer-active dimer equilibrium was present. The 5--6-fold activation by bovine serum albumin required the presence of free thiol groups in the albumin and involved association of albumin with the enzyme.

Animals↗
PubMed1974

Effect of sodium nitrite on toxin production by Clostridium botulinum in bacon.

Pork bellies were formulated to 0, 30, 60, 120, 170, or 340 mug of nitrite per g of meat and inoculated with Clostridium botulinum via pickle or after processing and slicing. Processed bacon was stored at 7 or 27 C and assayed for nitrite, nitrate, and botulinal toxin at different intervals. Nitrite levels declined during processing and storage. The rate of decrease was more rapid at 27 than at 7 C. Although not added to the system, nitrate was detected in samples during processing and storage at 7 and 27 C. The amount of nitrate found was related to formulated nitrite levels. No toxin was found in samples incubated at 7 C throughout the 84-day test period. At 27 C, via pickle, inoculated samples with low inoculum (210 C. botulinum per g before processing and 52 per g after processing) became toxic if formulated with 120 mug of nitrite per g of meat or less. Toxin was not detected in bacon formulated with 170 or 340 mug of nitrite per g of meat under these same conditions. Toxin was detected at all formulated nitrite levels in bacon inoculated via the pickle with 19,000 C. botulinum per g (4,300 per g after processing) and in samples inoculated after slicing. However, increased levels of formulated nitrite decreased the probability of botulinal toxin formation in bacon inoculated by both methods.

Animals↗
PubMed1973

Effect of nitrite and nitrate on toxin production by Clostridium botulinum and on nitrosamine formation in perishable canned comminuted cured meat.

Comminuted ham was formulated with different levels of sodium nitrite and nitrate, inoculated with Clostridium botulinum, and pasteurized to an internal temperature of 68.5 C. When added to the meat, nitrite concentrations decreased, and cooking had little effect on them. Nitrite concentrations decreased more rapidly during storage at 27 than at 7 C; however they remained rather constant at formulated levels throughout the experiment at both incubation temperatures. The level of nitrite added to the meat greatly influenced growth and toxin production of C. botulinum. The concentration of nitrite necessary to effect complete inhibition was dependent on the inoculum level. With 90 C. botulinum spores/g of meat, botulinum toxin developed in samples formulated with 150 but not with 200 mug of nitrite per g of meat. At a spore level of 5,000/g, toxin was detected in samples with 400 but not with 500 mug of nitrite per g of the product incubated at 27 C. At lower concentrations of nitrite, growth was retarded at both spore levels. No toxin developed in samples incubated at 7 C. Nitrate showed a statistically significant inhibitory effect at a given nitrite level; however, the effect was insufficient to be of practical value. Analyses for 14 volatile nitrosamines from samples made with varying levels of nitrite and nitrate were negative at a detection level of 0.01 mug of nitrite or nitrate per g of meat.

Animals↗
PubMed1973

Effect of sodium nitrite and sodium nitrate on botulinal toxin production and nitrosamine formation in wieners.

Wieners were formulated and processed approximating commercial conditions as closely as possible. Twenty-four batches of product were made with the addition of six levels of sodium nitrite (0, 50, 100, 150, 200, and 300 mug/g), four levels of sodium nitrate (0, 50, 150, and 450 mug/g), and two levels of Clostridium botulinum (0 and 620 spores/g). After formulation, processing, and vacuum packaging, portions of each batch were incubated at 27 C or held for 21 days at 7 C followed by incubation at 27 C for 56 days. The latter storage condition approximated distribution of product through commercial channels and potential temperature abuse at the consumer level. Samples were analyzed for botulinal toxin, nitrite, and nitrate levels after 3, 7, 14, 21, 28, and 56 days of incubation. When nitrite was not added, toxic samples were detected after 14 days of incubation at 27 C. At the lowest level of nitrite added (50 mug/g), no toxic samples were observed until 56 days of incubation. Higher levels of nitrite completely inhibited toxin production throughout the incubation period. Nine uninoculated samples, representing various levels and combinations of nitrite and nitrate, were evaluated organoleptically. The flavor quality of wieners made with nitrite was judged significantly higher (P = 0.05) than of wieners made without nitrite. The nine samples were negative for 14 volatile nitrosamines at a sensitivity level of 10 ng/g. The results indicated that nitrite effectively inhibited botulinal toxin formation at commercially employed levels in wieners and that detectable quantities of nitrosamines were not produced during preparation and processing of the product for consumption.

Botulinum Toxins↗