Plasmid encoded β-lactamases resistant to inhibition by clavulanic acid produced by calf faecal coliforms

Two new plasmid encoded β-lactamase enzymes produced by a strain of Escherichia coli and a strain of Citrobacter freundii isolated from calf faeces have been characterised. Both enzymes were similar to TEM-1 in terms of substrate and inhibition profiles and physical properties but differed from TEM-1 in being far less susceptible to the β-lactamase inhibitors clavulanic acid or tazobactam. In each case transfer of the the plasmid E coli K12 rendered it clinically resistant to the combination of amoxycillin and clavulanic acid. The β-lactamase from the E coli had an iso-electric point (pI) of 5·4 and was encoded on a plasmid of 95 Kbp which also mediated resistance to tetracycline, sulphonamides, apramycin, streptomycin and gentamicin. The β-lactamase from the C freundii had a pI of 5·2 and was encoded on a 75 Kbp plasmid which also mediated resistance to trimethoprim, chloramphenicol, apramycin, gentamicin and tobramycin.     

Pathogenicity of recent isolates of infectious bursal disease virus in specific-pathogen-free chickens: protection conferred by an intermediate vaccine strain

The pathogenicity of recent isolates of infectious bursal disease virus and the protection conferred against them by a commercial vaccine strain of intermediate virulence were examined in specific-pathogen-free chickens. Based on clinical signs, mortality, and macroscopic lesions in susceptible chickens, the isolates designated as A-Delmarva and U-28 were distinct from a previously known serotype I virulent isolate (Edgar). Histopathological analysis of the bursa of Fabricius did not establish differences between the field isolates. Although the vaccine strain produced some degree of bursal damage in antibody-free chickens, it was significantly less severe than the damage caused by the field isolates. The active immune response induced by vaccination was cross-protective against the pathological effects produced by the different isolates used in this study.     

Strangles in horse studs: Incidence, risk factors and effect of vaccination

A questionnaire survey of 179 horse studs in New South Wales was conducted to estimate the incidence of strangles during 1985 to 1988, to identify risk factors for strangles outbreaks and to assess the effect of strangles vaccination. Forty-nine of the studs (27.4%) had at least one strangles outbreak during this period and 62 studs (34.6%) had at least one case of strangles. The average incidence of strangles was 2.1 cases per 100 horses per year. The risk of strangles increased progressively with the total horse population and rose markedly when more than 100 mares had been served in the 1988-89 season. Certain types of feeders, fences and water sources were also significantly associated with outbreaks of strangles. Strangles vaccine was used on 63 studs (35.2%). Thirty-seven of these (58.7%) used the manufacturer's recommended vaccination regime. When other risk factors were taken into account, vaccination had no significant effect on the likelihood of a strangles outbreak.     

Comparison of colloid,thyroid follicular epithelium,and thyroid hormone concentrations in healthy and severely sick dogs

OBJECTIVES: To compare serum concentrations of total thyroxine (TT4), free thyroxine (fT4), and thyroid-stimulating hormone (TSH), as well as measures of thyroid follicular colloid and epithelium, between groups of healthy dogs and severely sick dogs. DESIGN: Cross-sectional study. ANIMALS: 61 healthy dogs and 66 severely sick dogs. PROCEDURE: Serum samples were obtained before euthanasia, and both thyroid lobes were removed immediately after euthanasia. Morphometric analyses were performed on each lobe, and serum TT4, fT4, and TSH concentrations were measured. RESULTS: In the sick group, serum TT4 and fT4 concentrations were less than reference range values in 39 (59%) and 21 (32%) dogs, respectively; only 5 (8%) dogs had high TSH concentrations. Mean serum TT4 and fT4 concentrations were significantly lower in the sick group, compared with the healthy group. In the healthy group, a significant negative correlation was found between volume percentage of colloid and TT4 or fT4 concentrations, and a significant positive correlation was found between volume percentage of follicular epithelium and TT4 or fT4 concentrations. A significant negative correlation was observed between volume percentages of colloid and follicular epithelium in both groups. CONCLUSIONS AND CLINICAL RELEVANCE: TT4 and fT4 concentrations are frequently less than reference range values in severely sick dogs. Therefore, thyroid status should not be evaluated during severe illness. The absence of any significant differences in mean volume percentages of follicular epithelium between healthy and severely sick dogs suggests that these 2 groups had similar potential for synthesizing and secreting thyroid hormones.     

Genotyping of Serratia marcescens Strains Associated with Cucurbit Yellow Vine Disease by Repetitive Elements-Based Polymerase Chain Reaction and DNA-DNA Hybridization

ABSTRACT The bacterium that causes cucurbit yellow vine disease (CYVD) has been placed in the species Serratia marcescens based on 16S rDNA and groE sequence analysis. However, phenotypic comparison of the organism with S. marcescens strains isolated from a variety of ecological niches showed significant heterogeneity. In this study, we compared the genomic DNA of S. marcescens strains from different niches as well as type strains of other Serratia spp. through repetitive elements-based polymerase chain reaction (rep-PCR) and DNA-DNA hybridization. With the former, CYVD strains showed identical banding patterns despite the fact that they were from different cucurbit hosts, geographic locations, and years of isolation. In the phylogenetic trees generated from rep-PCR banding patterns, CYVD strains clearly were differentiated from other strains but formed a loosely related group with S. marcescens strains from other niches. The homogeneity of CYVD strains was supported further by the DNA relatedness study, in that labeled DNA from the cantaloupe isolate, C01-A, showed an average relative binding ratio (RBR) of 99%, and 0.33% divergence to other CYVD strains. Used as a representative strain of CYVD, the labeled C01-A had a RBR of 76%, and a 4.5% divergence to the S. marcescens type strain. These data confirm the previous placement of CYVD strains in S. marcescens. Our investigations, including rep-PCR, DNA-DNA hybridization, and previous phenotyping experiments, have demonstrated that CYVD-associated strains of S. marcescens cluster together in a group significantly different from other strains of the species.     

Identification, Phylogenetic Analysis, and Biological Characterization of Serratia marcescens Strains Causing Cucurbit Yellow Vine Disease

ABSTRACT A serious vine decline of cucurbits known as cucurbit yellow vine disease (CYVD) is caused by rod-shaped bacteria that colonize the phloem elements. Sequence analysis of a CYVD-specific polymerase chain reaction (PCR)-amplified 16S rDNA product showed the microbe to be a gamma-proteobacterium related to the genus Serratia. To identify and characterize the bacteria, one strain each from watermelon and zucchini and several noncucurbit-derived reference strains were subjected to sequence analysis and biological function assays. Taxonomic and phylogenetic placement was investigated by analysis of the groE and 16S rDNA regions, which were amplified by PCR and directly sequenced. For comparison, eight other bacterial strains identified by others as Serratia spp. also were sequenced. These sequences clearly identified the CYVD strains as Serratia marcescens. However, evaluation of metabolic and biochemical features revealed that cucurbit-derived strains of S. marcescens differ substantially from strains of the same species isolated from other environmental niches. Cucurbit strains formed a distinct cluster, separate from other strains, when their fatty acid methyl ester profiles were analyzed. In substrate utilization assays (BIOLOG, Vitek, and API 20E), the CYVD strains lacked a number of metabolic functions characteristic for S. marcescens, failing to catabolize 25 to 30 compounds that were utilized by S. marcescens reference strains. These biological differences may reflect gene loss or repression that occurred as the bacterium adapted to life as an intracellular parasite and plant pathogen.