No abstract

Summary A total of 70 puppies and their dams, distributed in 14 litters, were submitted to weekly fecal examinations for C. difficile during the first 10 weeks after birth. During the study, 94.3% of the puppies and 42.9% of the dams harboured C. difficile at least once in their feces. We calculated that 58% of the puppies carried toxigenic C. difficile at least once during the survey. In the puppies, C. difficile carriage rates ranging from 3.1% to 67.1% were observed at different moments of the observation period. In comparison, C. difficile carriage rate was 1.4% in a control group of healthy dogs more than 3 months old. Discrepancies in the toxigenic phenotype of the C. difficile strains isolated in the same litter showed that the neonate dogs were transiently infected with different strains, and that the dam is often not the source of infection with C. difficile. We could not demonstrate any pathogenicity of C. difficile for neonate dogs.

A new miniaturized 4-h method for the identification of anaerobic bacteria, ATB 32 A (API System SA, Montalieu Vercieu, France), was evaluated against conventional methods of identification. The evaluation was done by using 260 recent clinical isolates and 21 reference strains of anaerobic bacteria. All reference strains were correctly identified and did not figure in the detailed analysis. Of the 140 gram-negative bacilli, 90.6% of Bacteroides spp. and 95.5% of Fusobacterium spp. were correctly identified to the species level, with an additional 8.4% of the Bacteroides spp. being identified to the genus level. Clostridia were correctly identified in 85.9% of cases, with an additional 9.9% being identified to the genus level. Peptostreptococci were correctly identified in 91.6% of cases. The 4 strains of Actinomyces spp. were ail identified correctly, as were 10 of the 11 strains of Propionibacterium spp. A total of 3.1% of strains were not identified by ATB 32 A, while for 1.9% of strains, completely false identifications were obtained. Estimation of the individual preformed enzyme results may pose problems, although these decrease with familiarity with the system. With certain enzyme profiles, additional testing was necessary to arrive at an identification; however, there was no requirement for gas-liquid chromatography. If certain additions are made to the data base and the difficulties of determination of organisms to the species level among the non-Bacteroides fragilis (sensu stricto) members of the B. fragilis group can be reduced, this system holds promise as a reliable standardized alternative for the identification of anaerobic bacteria in clinical laboratories.

Immunoadsorbers based on 2.0 x 6.0 mm i.d., epoxy-bearing, methacrylate-based monolithic disks were developed in order to target myoglobin and N-terminal pro-natriuretic peptide (NT-proBNP), two biomarkers involved in cardiovascular disease. In both cases, antibodies were successfully coupled to the polymeric disk material. The developed immunoadsorbers permitted the selective isolation of myoglobin and NT-proBNP from human serum. Myoglobin was successfully isolated and detected from serum samples at concentrations down to 250 fmol microL(-1). However, the affinity of the antibodies was not sufficient for the analysis of low-concentration clinical samples. Frontal analysis of anti-NT-proBNP disks revealed the ability of the immunoadsorber to bind up to 250 pmol NT-proBNP, which is more than sufficient for the analysis of clinical samples. Anti-NT-proBNP disks showed good stability over more than 18 months and excellent batch-to-batch reproducibility. Moreover, anti-NT-proBNP disks permitted the isolation of NT-proBNP at concentrations down to 750 amol microL(-1) in serum, corresponding to concentrations of strongly diseased patients. Using reversed-phase trapping columns, the detection of NT-proBNP eluted from immunoadsorbers by mass spectrometry was achieved for concentrations down to 7.8 fmol microL(-1).

Development of an avidinbiotin dot enzyme-linked immunosorbent assay and its comparison with other serological tests for diagnosis of glanders in equines. Vet Microbiol 15:77-85.