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Biomedical subjects

J E Moore

Publications and source records attributed to J E Moore.

At least 73 records · Page 4Linked to original sources

Novel employment of lactate dehydrogenase release from porcine aortic endothelial cells (PAEC) as a quantitative marker of cytotoxic activity in thermophilic Campylobacter spp. from human faecal isolates, poultry and environmental sources.

The aim of this study was to employ a novel cytotoxicity assay based on primary porcine aortic endothelial cells in combination with a lactate dehydrogenase release assay to quantitatively determine differences in cytotoxin production between Campylobacter jejuni, C. coli, C. lari and urease-positive thermophilic campylobacters (UPTC), isolated from human faeces, animals and environmental sources. Campylobacter isolates totalling 34 and comprising of C. jejuni (n = 24) C. coli (n = 5) and UPTC (n = 4) and C. lari (n = 1) were analysed. The cytotoxic response ranged from 32.15 to 64.47% and 33.08 to 59.41%, for C. jejuni from chicken and human isolates, respectively and there was no statistically significant difference (P > 0.05) in cytotoxic response between C. jejuni isolated from humans and chicken isolates (50.78% versus 50.55% cytotoxicity, respectively). However, there was a difference in response between C. jejuni and C. coli isolated from chickens (50.78% versus 33.22% cytotoxicity, respectively). The greatest cytotoxic response was obtained with the UPTC group of organisms examined (n = 4 isolates) (mean cytotoxic response = 57.11% cytotoxicity. Employment of this cytotoxin assay may help identify virulent strains in poultry that could potentially proceed to cause clinical problems for humans and thus intervention measures targeted at the reduction or elimination of such specific strains, may be sought.

Animals↗

Haemophilus segnis: a rare cause of endocarditis.

This report presents a case of endocarditis due to Haemophilus segnis, which represents a speciation difficulty for the routine laboratory. In this study, a molecular approach provided speciation, which was confirmed phenotypically by a reference laboratory. The use of molecular genotypic analysis is an additional strategy in the investigation of endocarditis. It has applications not only in isolate identification but also in primary detection of infection, particularly in patients whose blood is culture negative by conventional methodologies.

Base Sequence↗

Prevalence of bacterial faecal pathogens in separated and unseparated stored pig slurry.

AIMS: To examine the prevalence and diversity of bacterial faecal pathogens in unseparated slurry, separated solids and liquid fractions from a commercial pig farm. METHODS: A total of 43 stored slurry specimens originating from a fattening house over the period February-April 2002 were analysed, consisting of unseparated (n = 14) slurry, separated solids (n = 16) and separated liquid (n = 13). Specimens were examined for the presence of five bacterial pathogens including Salmonella spp., Shigella spp., Campylobacter spp., Escherichia coli O157 and Yersinia enterocolitica. Selective enrichment and plating methods were employed for detection of Salmonella spp. and Campylobacter spp. and conventional selective plating techniques for the remaining genera. Antibiogram profiles to 12 antibiotic agents were obtained for all Salmonella isolates obtained. RESULTS: Salmonella spp. were identified in all components of the slurry specimens, whereas Campylobacter spp. was only recovered from the unseparated and separated liquid fractions. In both cases, the separated liquid fraction had the highest prevalence of pathogens and the separated solid fraction had the lowest prevalence. None of the slurry specimens examined were positive for E. coli O157:H7, Shigella spp. or Y. enterocolitica. Twenty-nine isolates of Salmonella were recovered from the slurry specimens, comprising seven serovars, of which Salmonella manhattan was the most prevalent, accounting for over half [15 of 29 (51.7%)] of all Salmonella isolates. Salmonella anatum, Salm. derby, Salm. give, Salm. heidelberg, Salm. simi and Salm. stanley serovars were also recovered. All Salmonella isolates were sensitive to ampicillin, augmentin (amoxicillin/clavulanic acid), chloramphenicol, ciprofloxacin, gentamicin, kanamycin and trimethoprim, but has variable resistance to tetracycline (100%), sulphonamides (84.6%), furazolidone (38.5%), nalidixic acid (15.4%) and streptomycin (15.4%). The majority (57.7%) of isolates displayed antibiotic resistance to at least two antibiotic agents, followed by 34.6% of isolates being resistant to three agents and the remainder (7.7%) being resistant to four antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated a marked reduction in the prevalence of Campylobacter and Salmonella in the solids component of separated pig slurry. The adoption of control processes such as aeration of slurry prior to its spread onto agricultural land and newer approaches to pathogen reduction should be investigated, to reduce the transmission of pathogens from pig slurry to the environment.

Animal Husbandry↗

Structural analysis and genetic variation of the 16S-23S rDNA internal spacer region from Micrococcus luteus strains.

AIMS: To clone and sequence the 16S-23S ribosomal DNA (rDNA) internal spacer region (ISR) from Micrococcus luteus. METHODS AND RESULTS: The primer pair for 16S-23S rDNA ISR amplified a fragment of about 850 bp in length for two strains, JCM3347 and JCM3348 and a fragment of about 790 bp for a strain, ATCC9341. After sequencing the ISRs were identified by the comparison of the ISRs and the flanking regions of ISR. CONCLUSIONS: Although the sequence difference of the ISR occurred at only one position between the two JCM strains, the highly variable length (440 and 370 bp) and sequence similarity (about 40%) were demonstrated between the ISRs of the two JCM strains and a ATCC strain. SIGNIFICANCE AND IMPACT OF THE STUDY: A CCTCCT sequence was first detected at the 3'-end of the 16S rDNA of the three strains. Moreover, highly similar sequence to the 21-bp region containing a putative rRNA processing site was observed in the ISR of the three strains. Interestingly, no intercistronic tRNAs were demonstrated in the ISRs from the three strains.

Base Sequence↗

A comparison of pulmonary exacerbations with single and multiple organisms in patients with cystic fibrosis and chronic Burkholderia cepacia infection.

STUDY OBJECTIVES: To compare pulmonary exacerbations with single and multiple organisms in patients with chronic Burkholderia cepacia infection. DESIGN: Data was collected over a 23-month period and for each of the patients two episodes of pulmonary exacerbation were identified, one with B. cepacia isolated as the sole respiratory pathogen and the other with one or more coinfecting organisms grown from the sputum culture. SETTING: Regional tertiary referral center for Respiratory Medicine and Adult Cystic Fibrosis Center for Northern Ireland, Belfast City Hospital. PATIENTS: Adult (over 16 years) CF patients with chronic (more than 4 years) B. cepacia complex infection who attended the Belfast City Hospital between January 1997 and November 1998. MEASUREMENTS: Forced expiratory volume in 1s, forced vital capacity, forced expiratory flow rate 25-75, C-reactive protein, leucocyte count and body mass index were measured before and after two weeks of treatment and analysed for any differences in change between the two episodes. RESULTS: All variables in both groups improved following treatment except for body mass index, which remained unchanged. No significant differences between the measurements were identified on comparison of single (monomicrobial) and multiple (polymicrobial) isolate episodes. CONCLUSION: Patients with chronic B. cepacia infection who develop a pulmonary exacerbation improve, as reflected by clinical and biochemical markers, following IV antibiotic treatment. Pulmonary exacerbations with multiple organisms are no more severe than those where only B. cepacia is isolated.

Adolescent↗

Comparison of three techniques for estimating the forage intake of lactating dairy cows on pasture.

Quantifying DMI is necessary for estimation of nutrient consumption by ruminants, but it is inherently difficult on grazed pastures and even more so when supplements are fed. Our objectives were to compare three methods of estimating forage DMI (inference from animal performance, evaluation from fecal output using a pulse-dose marker, and estimation from herbage disappearance methods) and to identify the most useful approach or combination of approaches for estimating pasture intake by lactating dairy cows. During three continuous 28-d periods in the winter season, Holstein cows (Bos taurus; n = 32) grazed a cool-season grass or a cool-season grass-clover mixture at two stocking rates (SR; 5 vs. 2.5 cows/ha) and were fed two rates of concentrate supplementation (CS; 1 kg of concentrate [as-fed] per 2.5 or 3.5 kg of milk produced). Animal response data used in computations for the animal performance method were obtained from the latter 14 d of each period. For the pulse-dose marker method, chromium-mordanted fiber was used. Pasture sampling to determine herbage disappearance was done weekly throughout the study. Forage DMI estimated by the animal performance method was different among periods (P < 0.001; 6.5, 6.4, and 9.6 kg/d for Periods 1, 2, and 3, respectively), between SR (P < 0.001; 8.7 [low SR] vs. 6.3 kg/d [high SR]) and between CS (P < 0.01; 8.4 [low CS] vs. 6.6 kg/d [high CS]). The period and SR effect seemed to be related to forage mass. The pulse-dose marker method generally provided greater estimates of forage DMI (as much as 11.0 kg/d more than the animal performance method) and was not correlated with the other methods. Estimates of forage DMI by the herbage disappearance method were correlated with the animal performance method. The difference between estimates from these two methods, ranging from -4.7 to 5.4 kg/d, were much lower than their difference from pulse-dose marker estimates. The results of this study suggest that, when appropriate for the research objectives, the animal performance or herbage disappearance methods may be useful and less costly alternatives to using the pulse-dose method.

Animal Feed↗

Pasture forages, supplementation rate, and stocking rate effects on dairy cow performance.

Objectives were to evaluate effects of forage species, stocking rate, and supplementation rate on performance and physiology of grazing lactating Holstein cows under intensive rotational stocking management during summer. Eight treatments were arranged in a 2 x 2 x 2 factorial design. Animals (n = 62) grazed pastures of Tifton 85 bermudagrass or Florigraze rhizoma peanut, a tropical legume. Low and high stocking rates were 7.5 and 10.0 cows/ha for bermudagrass and 5.0 and 7.5 cows/ha for rhizoma peanut. Within each forage-stocking rate combination, cows were fed supplement at 0.33 or 0.5 kg of supplement (as-fed basis)/kg daily milk production. Cows grazing rhizoma peanut pastures produced more milk (16.9 vs. 15.4 kg/d) but had higher rectal temperatures (39.4 vs. 39.1 degrees C). Milk production per cow was improved at the higher stocking rate for bermudagrass but was reduced at the higher stocking rate for peanuts. Increasing supplementation rate boosted plasma glucose, milk production, and milk protein percent. Increased supplementation rate had a greater positive impact on milk production of cows grazing bermudagrass compared to rhizoma peanut (21.9 vs. 10.6% increase) due to a lower substitution of grain for forage intake. Organic matter intakes of forage, supplement, and total diet were greatest by cows grazing rhizoma peanut pastures and averaged 12.4, 6.1, and 18.5 kg/d compared to 9.2, 5.4, and 14.6 kg/d for cows grazing bermudagrass. Despite lower individual feed intake and performance, production per unit land area was 29% greater (112 vs. 90 kg of milk/ha per d) for cows grazing bermudagrass due to the greater stocking rate possible with that forage. Only cows supplemented at the high rate and kept at the high stocking rate on bermudagrass maintained body weight. Cows on other treatments lost body weight. Tifton 85 bermudagrass appears to be an excellent summer forage for dairy cows grazing in the southeastern U.S. given its nutritive value characteristics and high yields. Optimum stocking rate may be as high as 10 cows/ha during times of peak growth of forage for low-to-moderately producing cows fed supplement. Furthermore, the positive milk production response to additional supplement when cows grazed Tifton 85 pastures (0.8 kg/kg of supplement), indicates the value of providing supplement to cows grazing this moderate quality forage.

Animal Nutritional Physiological Phenomena↗

Biohazards and ecotoxicological considerations of landspreading of spent compost wastes.

Spent mushroom compost (SMC) is a major waste of the mushroom industry with low economic value. SMC arises after mushroom production in phase II compost (pIIC), predominantly comprising straw and chicken litter as principal raw ingredients. The majority of SMC waste is disposed off by application to agricultural land. It is an attractive proposition for utilising SMC as soil inorganic fertiliser supplementation. However, there is limited data available as to the consequences of this method of disposal either in terms of microbiological loading of food-borne pathogens and those of concern to mushroom industry itself. The resulting imbalance of the natural flora of the agricultural land has not been properly audited. This study aims to initially examine SMC for prevalence of faecal bacterial pathogens including Campylobacter spp., Salmonella spp. and Listeria monocytogenes that may arise from chicken litter. At another level, it aims to ascertain the pathogenic bacteria (Pseudomonas syringae, pv phaseolicola or tolasii) and fungal populations (Trichoderma, Verticillium species) originating mainly from the straw component of the SMC, which are of concern to the mushroom industry. Lastly, the study would also qualitatively identify the diversity of bacterial populations within SMC. This was largely accomplished through employment of rDNA, PCR and direct sequencing strategies on the culturable microflora. However, for specific mushroom pathogens, nucleic acids (DNA or RNA) were directly extracted from composts before subjecting to sequence analysis. In accordance with the current legislation (ABP 02/02, Animal By Products wastes disposal EC No. 1774/2002), it is imperative to regulate the farm wastes carrying residues from animal sources including SMC before they are regarded safe for land spreading operations. The ecological microbe-microbe and plant-microbe interactions that potentially occur between the native bacterial soil flora and those added annually (approximately 10(18) cells) needs to be reviewed with caution. The above study highlights the ecological consequences involved in the disposal of SMC wastes on agricultural land and its implications for plant, animal and human health.

Agaricales↗

Molecular genotyping of isolates of Actinobacillus actinomycetemcomitans by pulsed-field gel electrophoresis after separate digestion with Sse8387I and XhoI.

Genomic DNA from 18 Japanese clinical isolates of Actinobacillus actinomycetemcomitans was obtained from six periodontitis patients and analyzed using pulsed-field gel electrophoresis (PFGE) after separate digestion with Sse8387I and with XhoI. Three isolates from an identical patient were found to share an identical PFGE profile, and isolates from distinct patients were found to have PFGE profiles distinctly different from each other. Consequently, the 18 Japanese clinical isolates were discriminated into six distinct genotypes by means of PFGE. The genomic DNA from the other six reference strains (ATCC33384, ATCC43717, ATCC 43718, JCM2434, JCM2435 and Nig-1) was discriminated into six genotypes by the same PFGE methodology, and these six genotypes were found to be distinctly different from the six genotypes of the 18 Japanese clinical isolates described above. Serotyping demonstrated three PFGE genotypes in the serotype a strains, four the serotype b strains and three the serotype c strains. The present results clearly suggest that the PFGE procedure after separate digestion with Sse8387I and with XhoI has an excellent discriminatory power amongst strains and has a good genotypability for A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗

Hemodynamics and wall mechanics of a compliance matching stent: in vitro and in vivo analysis.

PURPOSE: Evidence is emerging that the abrupt compliance mismatch that exists at the junction between the stent ends and the host arterial wall disturbs both the vascular hemodynamics and the natural wall stress distribution. These stent-induced alterations are greatly reduced by smoothing the compliance mismatch between the stent and host vessel. A stent that provides this smooth transition in compliance, the compliance matching stent (CMS), has been developed. This study attempts to evaluate the hemodynamics and wall mechanical consequences of the CMS both in vitro and in vivo. MATERIALS AND METHODS: Finite element analysis was used to assess the solid mechanical behavior (compliance and stress) of the CMS in a stent/artery hybrid structure. A similar analysis was performed with a Palmaz stent. In vivo hemodynamics and wall mechanical changes induced by the CMS were investigated in a swine model from direct measurements of flow, pressure, diameter, and histology in the stented segment of superficial femoral arteries after 7 days. RESULTS: Finite element analysis showed that the abrupt compliance mismatch was substantially smoothed between the vessel portions with and without the stent with CMS segments. Circumferential stress was also markedly reduced with the CMS compared to other stent. The in vivo results showed that the CMS was efficient in compliance matching and did not dampen flow or pressure waves downstream the stent. Concurrent histology showed limited thrombus and inflammatory cell accumulation around the stent struts. CONCLUSION: These results indicate that the stent/artery hybrid structure can be compliance matched with proper stent design and that this structure limits solid mechanical stress and hemodynamic disturbances. It remains to be seen whether compliance-matched vascular stents reduce in-stent restenosis.

Animals↗

Subspecies characterization of porcine Campylobacter coli and Campylobacter jejuni by multilocus enzyme electrophoresis typing.

Enteropathogenic Campylobacterjejuni, C. coli and C. lari are currently the most common causes of acute infectious diarrhoeal illness in the UK. Many domestic animals, including pigs, act as natural reservoirs of these organisms and infection may occur through the ingestion of contaminated foodstuffs. C jejuni and C. coli, isolated from the livers of bacon pigs, were examined at subspecies level by multilocus enzyme electrophoresis (MEE) typing with seven enzymic loci. Polymorphological variation was highest with indophenol oxidase, isocitrate dehydrogenase and L-phenylalanyl-L-leucine peptidase giving 5. 5 and 4 alleles at these loci, respectively. The 35 Campylobacter isolates examined in this study (12 C. jejuni and 23 C coli) represented 30 unique electrophoretic types (ETs). Of these ETs, 8 unique types were detected for the 12 C jejuni isolates and 19 unique ETs were detected for the 23 C coli isolates. In addition, 3 types (ETs 2, 5, 10) were shared in common among C. jejuni and C coli. The average number of alleles per enzyme locus was 3.28. The mean genetic diversity, i.e. arithmetic average over all loci assayed, including monomorphic values, was 0.5573 and 0.5350 for C jejuni and C coli. respectively. Alleles were shared by C jejuni and C coli, suggesting an exchange of genetic material between the species. MEE analyses of isolates showed that there was a wide range of subspecies types within both C. jejuni and C coli in porcine livers. In certain cases, up to four phenotypically different strains of C coli were isolated from one liver, indicating multiple infections.

Animals↗