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Biomedical subjects

D G White

Publications and source records attributed to D G White.

At least 55 records · Page 3Linked to original sources

Chromobacteriosis in a Chinese red panda (Ailurus fulgens styani).

An adult Chinese red panda (Ailurus fulgens styani) transported by airplane from Florida to a North Dakota zoo died 1 week after arrival. Grossly, an interscapular abscess, subcutaneous inflammation, lymphadenitis, and pulmonary abscesses were observed. Microscopic findings included necrotizing inflammation in liver, lung, lymph node, and spleen. Chromobacterium violaceum was cultured from the interscapular abscess, liver, lung, and spleen and was injected into Swiss Webster mice. These mice died 18 hours postinoculation, and C. violaceum was cultured from liver, lung, and spleen. Chromobacterium violaceum is a sporadically reported but highly virulent pathogenic bacterium of both animals and humans typically found as a soil and water inhabitant of tropical and subtropical regions.

Animals↗

Abdominal distension in collapsed diarrhoeic calves: biochemical findings and treatment.

Thirty-seven of 53 diarrhoeic calves hospitalised for intravenous fluid therapy were classified as very severely acidotic (total carbon dioxide less than 8 mmol/litre) by using a Harleco apparatus. All the calves were given intravenously 10 to 20 litres of electrolyte solution which contained 144 mmol/litre sodium, 4 mmol/litre potassium, 113 mmol/litre chloride and 35 mmol/litre bicarbonate, and in addition the 37 very severely acidotic calves received 400 ml of 1M sodium bicarbonate in the first 5 litres of fluid administered. Sixteen of the 37 very severely acidotic calves had a distended right flank, suggesting the presence of a dilated fluid-filled viscus. Neither their history nor other clinical signs were useful predictors of the distension. The distended calves had significantly higher plasma concentrations of sodium and chloride, and significantly lower plasma creatinine concentrations than the calves which were not distended. Treatment was successful in all the 21 non-distended calves but four of the distended calves died despite treatment. The resolution of the distension in the successfully treated calves, coincided with a significant increase in plasma bicarbonate concentration and the passage of large amounts of malodorous mucoid faeces.

Abomasum↗

Characterization of eae+ Escherichia coli isolated from healthy and diarrheic calves.

Strains of Escherichia coli from 101 healthy and 114 diarrheic calves were screened by PCR for the eae (intimin) gene and Shiga toxin genes (stx). Each eae+ and eae/stx+ strain was examined for antimicrobial susceptibility, enterohemolysin activity, and the somatic O antigen was determined. An immunoassay was used to detect Shiga toxin antigens for the eae/stx+ E. coli. Significantly more (p = 0.005) of the healthy calves carried eae+ and eae/stx+ E. coli in their feces when compared to strains from diarrheic calves. Moreover, Shiga toxin antigens were detected significantly more (p = 0.001) often among the eae/stx+ strains from healthy calves when compared to eae/stx+ strains from diarrheic calves. However, significantly more (p = 0.001) of the eae+ and eae/stx+ strains from diarrheic calves were resistant to at least one of the antimicrobials tested, and the strains from diarrheic calves had a significantly (p = 0.05) higher rate of antimicrobial resistance to at least two different antimicrobial classes. No significant difference (p> or =0.05) was detected among the eae+ and eae/stx+ strains from healthy and diarrheic calves for enterohemolysin production. Serogroups O-negative, O5, O26, and O111 were predominate among both healthy and diarrheic calves.

Adhesins, Bacterial↗

Characterization of expanded-spectrum cephalosporin resistance in E. coli isolates associated with bovine calf diarrhoeal disease.

Antibiotic resistance among Escherichia coli isolates from diarrhoeal disease in cattle was studied. Many of the isolates were multiply resistant to beta-lactams, including expanded-spectrum cephalosporins, aminoglycosides, sulphonamides, tetracycline and fluoroquinolones. In many of the isolates, IEF revealed a strong beta-lactamase band compatible with overexpression of the AmpC beta-lactamase, either alone or in addition to TEM-type enzymes. Several of the isolates also possessed genes encoding virulence factors associated with animal and human diarrhoeal diseases. These results suggest that the use of antibiotics in animals could lead to a reservoir of antibiotic-resistant bacteria that could potentially infect humans.

Animals↗

Incidence and characterization of integrons, genetic elements mediating multiple-drug resistance, in avian Escherichia coli.

Antibiotic resistance among avian bacterial isolates is common and is of great concern to the poultry industry. Approximately 36% (n = 100) of avian, pathogenic Escherichia coli isolates obtained from diseased poultry exhibited multiple-antibiotic resistance to tetracycline, oxytetracycline, streptomycin, sulfonamides, and gentamicin. Clinical avian E. coli isolates were further screened for the presence of markers for class 1 integrons, the integron recombinase intI1 and the quaternary ammonium resistance gene qacEDelta1, in order to determine the contribution of integrons to the observed multiple-antibiotic resistance phenotypes. Sixty-three percent of the clinical isolates were positive for the class 1 integron markers intI1 and qacEDelta1. PCR analysis with the conserved class 1 integron primers yielded amplicons of approximately 1 kb from E. coli isolates positive for intI1 and qacEDelta1. These PCR amplicons contained the spectinomycin-streptomycin resistance gene aadA1. Further characterization of the identified integrons revealed that many were part of the transposon Tn21, a genetic element that encodes both antibiotic resistance and heavy-metal resistance to mercuric compounds. Fifty percent of the clinical isolates positive for the integron marker gene intI1 as well as for the qacEDelta1 and aadA1 cassettes also contained the mercury reductase gene merA. The correlation between the presence of the merA gene with that of the integrase and antibiotic resistance genes suggests that these integrons are located in Tn21. The presence of these elements among avian E. coli isolates of diverse genetic makeup as well as in Salmonella suggests the mobility of Tn21 among pathogens in humans as well as poultry.

Animals↗

Self-collection of group B Streptococcus cultures in pregnant women.

OBJECTIVE: This study assesses the sensitivity of self-collected rectovaginal culture specimens for group B Streptococcus by pregnant patients. METHODS: A volunteer sample of 240 pregnant women at 28 weeks gestation self-collected rectovaginal culture swabs to screen for the presence of group B Streptococcus. The patients' physicians collected second specimens for comparison. RESULTS: Twenty-four of 240 women grew group B Streptococcus on at least one culture (incidence, 10%). Twenty physician-collected specimens and 19 patient-collected specimens were positive (83 and 79% sensitivity, respectively). Fifteen patients (62.5%) had both physician-collected and patient-collected cultures grow group B Streptococcus. Cohen's kappa (kappa = 0.75) indicates a high degree of agreement between patient-collected and physician-collected cultures. CONCLUSIONS: Pregnant women are as likely as their attending physicians to obtain positive cultures for group B Streptococcus by self-collection of rectovaginal swabs.

Adult↗

Comparison of danofloxacin with baquiloprim/sulphadimidine for the treatment of experimentally induced Escherichia coli diarrhoea in calves.

Thirty-eight, one- to two-week-old calves with experimentally induced Escherichia coli diarrhoea were randomly assigned to three treatment groups. Two groups of 15 calves were treated intramuscularly once daily for three days with either danofloxacin mesylate at 1.25 mg/kg bodyweight, or with baquiloprim/sulphadimidine as a positive control (10 mg of combined active ingredient/kg); eight calves were treated with 0.9 per cent sodium chloride solution as a negative control (1 ml/20 kg). Faecal consistency, demeanour, hydration status, appetite and bodyweight were monitored before, during, and for four days after treatment by an investigator unaware of the animals' treatment. Before treatment, the clinical, biochemical, and faecal indices were similar among the groups. By 24 hours after treatment began, the proportion of observations of faeces recorded as of normal consistency was highest in the danofloxacin-treated group (26 of 60), compared with 16 of 60 in the baquiloprim/sulphadimidine treated groups and four of 32 in the control group. The proportion of calves with a normal demeanour was highest in the danofloxacin-treated group at all the evaluations and these calves gained significantly (P < 0.05) more weight (1.6 [0.27] kg) than the calves treated with baquiloprim/sulphadimidine (0.67 [0.36] kg). The calves in the danofloxacin-treated group maintained relatively normal blood pH values, whereas the calves in the control group became progressively acidotic. By the end of treatment, the mean bicarbonate concentration was significantly (P < 0.05) higher in the danofloxacin-treated calves than in the control group. The pH of the calves in the baquiloprim/sulphadimidine-treated group changed little during treatment, but by three days after the last treatment their mean pH had dropped to the level of the calves in the control group. The mean bicarbonate concentration of the baquiloprim/sulphadimidine-treated calves, like that of the danofloxacin-treated calves, was significantly (P < 0.05) higher than that of the calves in the control group.

Animals↗

Multidrug resistance following expression of the Escherichia coli marA gene in Mycobacterium smegmatis.

Expression of the Escherichia coli multiple antibiotic resistance marA gene cloned in Mycobacterium smegmatis produced increased resistance to multiple antimicrobial agents, including rifampin, isoniazid, ethambutol, tetracycline, and chloramphenicol. Cloned marR or marA cloned in the antisense direction had no effect. Resistance changes were lost with spontaneous loss of the plasmid bearing marA. A MarA mutant protein, having an insertional mutation within either of its two alpha-helices of the first putative helix-turn-helix domain, failed to produce the multiresistance phenotype in E. coli and M. smegmatis, indicating that this region is critical for MarA function. These results strongly suggest that E. coli marA functions in M. smegmatis and that a mar-like regulatory system exists in this organism.

Bacterial Proteins↗

Evaluation of a glutamine-containing oral rehydration solution for the treatment of calf diarrhoea using an Escherichia coli model.

A high-calorie oral rehydration solution (ORS) with glutamine (n=11) was more effective in correcting plasma, extracellular fluid and blood volume than solutions without (one WHO-type solution, n=6, and two high-glucose but glutamine-free solutions, n=7, n=12). It was the only solution to improve plasma volume significantly within 48 h and sustain the improvement throughout treatment; similarly, it was the only solution to correct packed-cell volume within 48 h and sustain the benefit to the end of treatment. At the end of treatment, the glutamine-treated calves were the only ones to avoid a significant weight loss compared with their pre-diarrhoeic values. The crucial difference between this solution and those used with glutamine previously is that it gave significant nutritional support whereas WHO type solutions did not. It also had more favourable effects on hyponatraemia and metabolic acidosis than a standard ORS. Use of a high-calorie ORS for 4 days (rather than 2 days of 50:50 admixture with milk replacer) brought additional beneficial effects on blood glucose and body weight.

Animals↗

Inhibition of the multiple antibiotic resistance (mar) operon in Escherichia coli by antisense DNA analogs.

The multiple antibiotic resistance operon (marORAB) in Escherichia coli controls intrinsic susceptibility and resistance to multiple, structurally different antibiotics and other noxious agents. A plasmid construct with marA cloned in the antisense direction reduced LacZ expression from a constitutively expressed marA::lacZ translational fusion and inhibited the induced expression of LacZ in cells bearing the wild-type repressed fusion. The marA antisense construction also decreased the multiple antibiotic resistance of a Mar mutant. Two antisense phosphorothioate oligonucleotides, one targeted to marO and the other targeted to marA of the mar operon, introduced by heat shock or electroporation reduced LacZ expression in the strain having the marA::lacZ fusion. One antisense oligonucleotide, tested against a Mar mutant of E. coli ML308-225, increased the bactericidal activity of norfloxacin. These studies demonstrate the efficacy of exogenously delivered antisense oligonucleotides targeted to the marRAB operon in inhibiting expression of this chromosomal regulatory locus.

Cloning, Molecular↗

Role of the acrAB locus in organic solvent tolerance mediated by expression of marA, soxS, or robA in Escherichia coli.

Escherichia coli K-12 strains are normally tolerant to n-hexane and susceptible to cyclohexane. Constitutive expression of marA of the multiple antibiotic resistance (mar) locus or of the soxS or robA gene product produced tolerance to cyclohexane. Inactivation of the mar locus or the robA locus, but not the soxRS locus, increased organic solvent susceptibility in the wild type and Mar mutants (to both n-hexane and cyclohexane). The organic solvent hypersusceptibility is a newly described phenotype for a robA-inactivated strain. Multicopy expression of mar, soxS, or robA induced cyclohexane tolerance in strains with a deleted or inactivated chromosomal mar, soxRS, or robA locus; thus, each transcriptional activator acts independently of the others. However, in a strain with 39 kb of chromosomal DNA, including the mar locus, deleted, only the multicopy complete mar locus, consisting of its two operons, produced cyclohexane tolerance. Deletion of acrAB from either wild-type E. coli K-12 or a Mar mutant resulted in loss of tolerance to both n-hexane and cyclohexane. Organic solvent tolerance mediated by mar, soxS, or robA was not restored in strains with acrAB deleted. These findings strongly suggest that active efflux specified by the acrAB locus is linked to intrinsic organic solvent tolerance and to tolerance mediated by the marA, soxS, or robA gene product in E. coli.

Bacterial Proteins↗

Growth of an Aspergillus flavus transformant expressing Escherichia coli beta-glucuronidase in maize kernels resistant to aflatoxin production.

Kernels of a maize inbred that demonstrated resistance to aflatoxin production in previous studies were inoculated with an Aspergillus flavus strain containing the Escherichia coli beta-D-glucuronidase reporter gene linked to a beta-tubulin gene promoter and assessed for both fungal growth and aflatoxin accumulation. Prior to inoculation, kernels were pin-wounded through the pericarp to the endosperm, pin-wounded in the embryo region, or left unwounded. After 7 days incubation with the fungus, beta-glucuronidase activity (fungal growth) in the kernels was quantified using a fluorogenic assay and aflatoxin B content of the same kernels was analyzed. Kernels of a susceptible inbred, similarly treated, served as controls. Results indicate a positive relationship between aflatoxin levels and the amount of fungal growth. However, resistant kernels wounded through the pericarp to the endosperm before inoculation supported an increase in aflatoxin B over levels observed in nonwounded kernels, without an increase in fungal growth. Wounding kernels of the resistant inbred through the embryo resulted in both the greatest fungal growth and the highest levels of aflatoxin B1 for this genotype. Maintenance of resistance to aflatoxin B1 in endosperm-wounded kernels may be due to the action of a mechanism which limits fungal access to the kernel embryo.

Aflatoxin B1↗

Marshmallows cause an outbreak of infection with Salmonella enteritidis phage type 4.

Thirty-six cases of Salmonella enteritidis phage type (PT) 4 infection, mainly in children, were notified in one local authority in the two weeks beginning on 23 October 1995. Twenty-four cases and 42 controls nominated by cases were included in a case control study, which showed a significant association between illness and the consumption of marshmallow confectionery from a bakery. S. enteritidis PT4 was isolated from samples of the marshmallow, and all isolates had the same plasmid and pulsed field gel electrophoresis profiles. The marshmallow had been made using raw egg white, which shows that not all caterers follow the Department of Health's advice.

Case-Control Studies↗

Chick cartilage fibronectin differs in structure from the fibronectin in limb mesenchyme.

Fibronectin, present in the extracellular matrix of several tissues, is heterogeneous in structure. This heterogeneity is largely due to the alternative splicing of three exons (IIIB, IIIA, and V) during processing of the fibronectin primary transcript. Previously, we determined that the splicing patterns of fibronectin mRNAs change from B+A+ to B+A- during the differentiation of mesenchyme into cartilage (V. D. Bennett, K. M. Pallante, and S. L. Adams, J. Biol. Chem. 266, 5918-5924, 1991). Therefore, the structure of fibronectin at the protein level most likely changes during chondrogenesis as well. In order to characterize the fibronectin protein in chick limb prechondrogenic mesenchyme and cartilage, we generated a polyclonal antibody specific for the region encoded by exon IIIB in the chick fibronectin gene. Immunoblot and immunohistochemistry analyses with this antibody and an antibody specific for the region encoded by exon IIIA indicate that both antibodies react with the fibronectin present in prechondrogenic mesenchyme. In contrast, only the exon IIIB antibody reacts with the fibronectin present in chick cartilage. Quantitative ELISA assays with these antibodies indicate that approximately 96% of the fibronectin in chick embryonic cartilage contains exon IIIB while less than 3% of the fibronectin contains exon IIIA. These results corroborate the structures of the fibronectin isoforms present in prechondrogenic mesenchyme and cartilage that were predicted from our previous characterization of the fibronectin mRNAs in these tissues and indicate that essentially all of the fibronectin in cartilage is synthesized and secreted by cartilage chondrocytes.

Alternative Splicing↗

Fallibility of faecal consistency as a criterion of success in the evaluation of oral fluid therapy for calf diarrhoea.

It is often said that the success of oral rehydration in humans depends on the adequacy of the improvement in the composition and volume of extracellular fluid, not reduction of faecal output. Indeed, the latter may increase initially. Such increases do not prevent the treatment from being effective but they may, falsely, undermine its acceptability to patients or those caring for them. This paper provides data to show that standard oral rehydration solutions used to treat experimentally induced calf diarrhoea procure identical improvements in plasma volume during the first 48 h, whether faeces improve or not, and those calves whose faecal consistency improved actually showed greater deterioration of extracellular fluid volume. While it is important for this to be appreciated by clinicians and explained to owners, it is absolutely imperative that those responsible for the approval of new therapeutic products for registration understand and accept that faecal consistency offers no reliable insight into the effectiveness of oral rehydration therapy for calf diarrhoea. It was, however, interesting that there was some relationship with correction of acidosis--perhaps because some of the contributing factors arise from colonic dysfunction.

Animals↗

Evaluation of a nutritive oral rehydration solution for the treatment of calf diarrhoea.

The essential constituents of a conventional oral rehydration solution (ORS) are sodium, glucose and a bicarbonate precursor. The glucose promotes sodium uptake but because these solutions are isotonic, it is insufficient to sustain calorie requirements. This paper examines the performance of a novel ORS with over three times the conventional glucose concentration, by comparing it with two leading commercial ORSs in calves with induced Escherichia coli diarrhoea. This solution showed greater ability than the current market-leading ORS to repair extracellular fluid and plasma volume and to correct both hyponatraemia and metabolic acidosis, especially in more severely affected calves. In acidotic calves it was more effective in correcting hyperkalaemia, probably by supplying glucose to promote cellular potassium uptake as well as by correcting the acidosis. It therefore appears possible to depart from the traditional isotonic formulations for calf ORSs and gain significant nutritional support while retaining effective rehydration and correction of acid-base and electrolyte disturbances. This seems especially important in young animals where energy deprivation imposes a particular penalty; the use of hypertonic ORSs should not, however, be extended to other species without further research.

Acid-Base Equilibrium↗

Multiple antibiotic resistance (mar) locus protects Escherichia coli from rapid cell killing by fluoroquinolones.

The multiple antibiotic resistance (mar) locus in Escherichia coli consists of two divergently expressed operons (marC and marRAB), both of which contribute to the Mar phenotype. Overexpression of the marRAB operon protected E. coli against rapid cell killing by fluoroquinolones. Inactivation of the operon in mar mutants restored a wild-type bactericidal susceptibility. Both operons of the locus were required for protection from the quinolone-mediated bactericidal activity in mar locus deletion mutants. The effect was lost at high concentrations of fluoroquinolones, unlike the case for the previously described genes hipA and hipQ. The inducible mar locus appears to specify a novel antibactericidal mechanism which may play a role in the emergence of fluoroquinolone-resistant clinical E. coli isolates.

Anti-Infective Agents↗

Escherichia coli cellulitis in broiler chickens: clonal relationships among strains and analysis of virulence-associated factors of isolates from diseased birds.

Thirty-nine Escherichia coli isolates from broiler chickens with cellulitis were serotyped and analyzed for clonal relationships by multilocus enzyme electrophoresis. The isolates were further characterized with respect to hemagglutination (HA); serum resistance; antibiotic susceptibility; production of aerobactin, colicin V, and hemolysin; expression of K1 or K5 capsule; sensitivity to cloacin DF13 after treatment with diphenylamine; expression of iron-regulated outer membrane proteins; and virulence in 1-day-old chickens. In addition, the isolates were examined for the presence of DNA sequences related to F1A (fim) and P (pap) fimbriae, aerobactin synthesis (iuc) and transport (iut), hemolysin operon hly, and TraT lipoprotein-induced serum resistance (traT). Only 38.4% of the isolates were typeable with standard O antisera, and of these, serogroups O25 and O78 were the most frequently observed. Multilocus enzyme electrophoresis, based on 20 enzymes, resolved 17 electrophoretic types, forming seven clusters. Isolates from four of these clusters fell into E. coli clone complexes that have been previously reported to be commonly associated with avian colibacillosis. All isolates expressed two to five iron-regulated outer membrane proteins, were resistant to serum and cloacin DF13, and possessed DNA sequences homologous to fim and iuc/iut. Most isolates (72%) were positive for traT, and a majority produced colicin V and aerobactin (92 and 82%, respectively). Assays for the presence of fim and pap DNA sequences, for HA, and for virulence gave variable results but suggest that cellulitis isolates may express F1A and/or other mannose-resistant HA fimbriae different from P and may be virulent in 1-day-old chickens. Our results support the hypothesis that cellulitis in broilers in many cases is caused by E. coli clones identical to other pathogenic avian E. coli strains. Certain clones may be specific to cellulitis, because 25% of the isolates tested belong to clusters not related to known clone complexes.

Animals↗