Competitive enzyme-linked immunosorbent assay for heartwater using monoclonal antibodies to a Cowdria ruminantium-specific 32-kilodalton protein

Hybridomas producing monoclonal antibodies (mAb) to Cowdria ruminantium were raised. Four mAbs of the IgG isotype reacted in western blots with a 32-kilodalton Cowdria protein (Cr32), which had previously been shown to be conserved and immunodominant. A fifth mAb of the IgM isotype recognized a 40-kDa Cowdria protein. The latter mAb was negative in an indirect fluorescent antibody test (IFA), whereas the other four were positive. mAb No. 4F10B4 showed the strongest signal in western blots using three different stocks of Cowdria. Immuno-gold labeling of Cowdria organisms in vitro using 4F10B4 showed that Cr32 has surface-exposed antigenic determinants. Using mAb 4F10B4, a competitive ELISA was developed which detected specific Cowdria antibodies in goat, sheep and cattle sera. Antibodies in animal sera competed with binding of mAb 4F10B4 to a crude sonicated Cowdria antigen obtained from infected endothelial cell cultures. The competition ELISA (CELISA) detected antibodies in 55 out of 70 (79%) goats experimentally infected with one of eight different Cowdria stocks. Fourteen out of the 15 sera which were shown negative in the CELISA were also negative in the IFA. Nevertheless, all 15 sera recognized some epitopes of the immunodominant Cowdria-specific 32 kDa protein as judged from their reaction with this protein in western blots. Overall, there was 89% agreement between CELISA and IFA considering all 70 goat sera. Moreover, antibodies were detected in nine out of nine sheep infected with one of three different stocks of Cowdria and in sera from calves experimentally infected by two different strains of heartwater. There were no cross-reactions with Ehrlichia phagocytophila antibodies in goat sera, nor with Anaplasma marginale antibodies in bovine sera. Lack of cross-reactivity and detection of antibodies to eight geographically widely distributed stocks of Cowdria, makes the competition ELISA a promising test for use in heartwater endemic areas.     

Influence of androgens on the genital tract of cyclic heifers

Cyclic heifers were implanted with the synthetic androgen trenbolone acetate (TBA = 17 beta-hydroxy-19-norandrosta-4, 9, 11-trien-3-on-17 beta-acetate, C2OH2403) (for formulas see Fig. 1). The influence of this androgen on the macroscopic and microscopic morphology of the genital tract was studied. The most striking result was the induction of polycystic ovaries. The clitoris was markedly enlarged and the cervix showed an increased amount of mucus. Microscopically, extensive folding of the cervical epithelium, consisting of tall swollen columnar cells with basal nuclei was seen. The vagina showed an increase of PAS-positive granules in the superficial layer of the epithelium, while there was no increase in the height of the epithelium. These effects are due to androgenic influence on the female genital tract and are in no case specific to one particular product. They could be of help in the detection of the illegal use of agents with androgenic activity.     

Serotypes of Erysipelothrix rhusiopathiae in Australian pigs, small ruminants, poultry, and captive wild birds and animals

Serotypes of 93 Australian isolates of Erysipelothrix rhusiopathiae from diseased domestic animals and poultry and a variety of captive wild birds and animals were determined by double diffusion gel precipitation. Two isolates, from the faeces of a swallow were also examined. Serotypes 1a, 1b and 2 were isolated from pigs and serotypes 1a, 1b, 2, 5, 15 and 21 from sheep or goats. Erysipelas in poultry was attributed to serotypes 1b, 5, 15 and 16. In captive wild birds serotypes 1b, 5, 6, 8, 14, 21 and an isolate reactive with antiserum to strain Seehecht were associated with septicaemic deaths. Single isolates from tissues of a bilby (Macrotis lagotis), black rat (Rattus rattus), brown snake (Pseudechis australis) and a bandicoot (Isoodon macrouris) were classified as serotypes 4, 4, 7, and 10 respectively. Six isolates were not able to be typed. Serotype 1b was the most widely distributed and most common (28%), being associated with disease in pigs, sheep, poultry and wild birds. Serotypes 1a or 2 were found in a more restricted range of animals, being commonly associated with erysipelas in pigs, less commonly in sheep and infrequently in other species. From diseased pigs, 26 of 33 isolates (79%) were serotypes 1a and 1b.     

Tetrachloroanisol: a source of musty taste in eggs and broilers

Ingestion by hens and broilers of specific chloroanisols present in some wood shavings used in poultry cages can result in a musty taste in poultry products.     

Investigation of changes in the middle latency auditory evoked potential during anesthesia with sevoflurane in dogs

OBJECTIVE: To investigate the middle latency auditory evoked potential (MLAEP) in awake dogs and dogs anesthetized with 2 concentrations of sevoflurane. ANIMALS: 10 adult Beagles. PROCEDURE: The MLAEP was recorded while dogs were awake and anesthetized with sevoflurane (end-tidal concentration, 2.7% or 3.5%).Three needle electrodes were inserted SC, and click stimuli were delivered biaurally. Signal acquisition, averaging, and analysis were performed by use of computer software developed in-house. Signals were recorded for 128 milliseconds, and the responses to 1,024 stimuli were averaged. Waveforms from 10 recordings in each dog were averaged, and latencies of peaks were measured. Data acquired for awake dogs and dogs anesthetized with high and low sevoflurane concentrations were compared statistically. RESULTS: Sevoflurane anesthesia attenuated the MLAEP so that only peaks P0, Na, and Pa could be identified. The MLAEP changes were maximal at the lower concentration of sevoflurane evaluated. The latencies of these peaks were significantly shorter in awake dogs, compared with values in anesthetized dogs. No difference in the peak latency was detected between the sevoflurane concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: In terms of CNS responsiveness, the effects of anesthesia with sevoflurane are similar to those of anesthesia with isoflurane. Data suggest that sevoflurane is not the inhalant agent of choice in a research setting where electroencephalographic measurements are to be recorded during anesthesia. The depression of the MLAEP waveform by sevoflurane also suggests that the MLAEP is not a suitable tool with which to monitor anesthetic depth during sevoflurane anesthesia in dogs.