Pulsed-field gel electrophoresis typing of Campylobacter fetus subsp. fetus isolated from sheep abortions in New Zealand

AIMS: To genotype Campylobacter fetus subsp. fetus isolates cultured from sheep abortions submitted to diagnostic laboratories in New Zealand during the year 2000 breeding season. To compare the types found nationally with those found in the Hawke' Bay region in 1999, and strains held in the New Zealand Reference Culture Collection, Medical Section (NZRM) from a study published in 1987.

METHODS: Campylobacter fetus subsp. fetus isolates cultured by veterinary diagnostic laboratories in the year 2000 breeding season, from sheep abortions from throughout New Zealand, were typed using pulsed-field gel electrophoresis (PFGE). In addition, seven freeze-dried C. fetus subsp. fetus isolates (strain numbers 2939–2945) from the NZRM, representing restriction types a–g found amongst sheep abortion isolates in a study published in 1987, were typed using PFGE.

RESULTS: In total, 293 C. fetus subsp. fetus isolates from 200 farms were obtained from veterinary diagnostic laboratories. Twenty-two distinct PFGE profiles were identified amongst the isolates. PFGE type B1 was predominant in each region of New Zealand and was identified from 66% of farms overall. Of the C. fetus subsp. fetus restriction types a–g lodged with the NZRM, 3/7 had PFGE profiles indistinguishable from profiles found in the current study. The other four restriction types had PFGE profiles that were unique but similar to those found in the current study.

CONCLUSIONS: PFGE type B1 was predominant amongst the C. fetus subsp. fetus isolates cultured from sheep abortions in each region of New Zealand in the year 2000, as was found in Hawke' Bay in 1999. The similarity between PFGE profiles of C. fetus subsp. fetus sheep abortion isolates from 1987 and 2000, and the relative prevalence of the PFGE groups, suggests that there has been no major genotypic shift in the population of C. fetus subsp. fetus implicated in sheep abortion in New Zealand during this time.     

Effects of Retinoids on the In Vitro Development of Capra Hircus Embryos to Blastocysts in Two Different Culture Systems

The objective of this study was to evaluate the effect of retinol (RT) and retinoic acid (RA) on the in vitro development of pre‐implantation goat embryos cultured in potassium simplex optimized medium or synthetic oviduct fluid or cocultured in oviductal cells monolayer either in potassium simplex optimized medium or synthetic oviduct fluid. A total of 2407 cumulus‐oocyte complexes were aspirated from 2 to 6 mm ovarian follicles from slaughtered animals. Selected cumulus‐oocyte complexes were subjected to in vitro maturation in TCM 199 for 24 h at 39°C in an atmosphere of 5% (v/v) CO₂ in humidified air. In vitro fertilization was performed in modified defined medium. Eighteen hours after in vitro fertilization, cumulus cells were removed and presumptive zygotes were randomly distributed into experimental groups. In Experiment 1, presumptive zygotes were cultured in potassium simplex optimized medium, potassium simplex optimized medium + RT, potassium simplex optimized medium + retinoic acid, synthetic oviduct fluid, synthetic oviduct fluid + RT and synthetic oviduct fluid + RA at 39°C in a humidified atmosphere of 5% (v/v) CO₂, 5% (v/v) O₂ and 90% (v/v) N₂. In Experiment 2, presumptive zygotes were cocultured in potassium simplex optimized medium + oviductal cells monolayer, potassium simplex optimized medium + RT + oviductal cells monolayer, potassium simplex optimized medium + RA + oviductal cells monolayer, synthetic oviduct fluid + oviductal cells monolayer, synthetic oviduct fluid + RT + oviductal cells monolayer and synthetic oviduct fluid + RA + oviductal cells monolayer in an atmosphere of 5% (v/v) CO₂ in humidified air. In both experiments, media were partially changed on day 2 after in vitro fertilization and unfertilized oocytes were excluded from the experiment. Embryos were cultured or cocultured for 8 days. In Experiment 1, there was no effect of RT or RA supplementation on the proportion of oocytes that reached the morula or blastocyst stages. By contrast, Experiment 2 demonstrated that the addition of 0.28 μg/ml RT and 0.5 μm RA to the embryo culture media stimulated (p < 0.05) development to the morula and blastocyst stages under the coculture conditions tested. In conclusion, retinoids play an important role in pre‐implantation development of goat embryos and can be used to enhance in vitro embryo production.     

Prevalence of Bartonella species,Rickettsia felis,haemoplasmas and the Ehrlichia group in the blood of cats and fleas in eastern Australia

Objectives To define the prevalence of Bartonella spp., Rickettsia felis, Mycoplasma haemofelis, ‘Candidatus Mycoplasma haemominutum’ (Mhm) and ‘Candidatus Mycoplasma turicensis’ (Mtc) in cats and their fleas in eastern Australia. Design and procedure Conventional PCR assays that detect Bartonella spp., M. haemofelis, Mhm, Mtc, Rickettsia spp., Ehrlichia spp., Anaplasma spp. and Neorickettsia spp. were performed on DNA extracted from blood and fleas collected from 111 cats. Cat sera were assayed by ELISA for IgG of Bartonella spp. Results DNA of M. haemofelis, Mtc and Mhm was amplified from 1 (0.9%), 1 (0.9%) and 17 cats (15.3%), respectively. Only DNA of Mhm was amplified from the 62 of 111 pooled flea samples (flea sets; 55.9%). Overall, the prevalence rates for Bartonella spp. DNA in the cats and the flea sets was 16.2% (18 cats) and 28.8% (32 flea sets), respectively. Bartonella spp. IgG was detected in 42 cats (37.8%), of which 11 (26.2%) were positive for Bartonella spp. DNA in their blood. R. felis DNA was amplified from 22 flea sets (19.8%), but not from cats. Overall, DNA of one or more of the organisms was amplified from 27% (30) of cats and 67.6% (75) of the flea sets. Conclusions This is the first Australian study to determine the prevalence of R. felis and B. clarridgeiae in both fleas and the cats from which they were collected. Flea-associated infectious agents are common in cats and fleas in eastern Australia and support the recommendation that stringent flea control be maintained on cats.     

Observations on diagnostic tests for paratuberculosis in a deer herd

Paratuberculosis was diagnosed in a herd of 62 fallow deer (Dama dama) and 38 sika deer (Cervus nipon) maintained in an animal park in Ohio. A comparison was made of delayed-type hypersensitivity responses, results of in vitro lymphocyte immunostimulation tests, and isolation of Mycobacterium paratuberculosis on fecal culture in 19 deer.     

Behavioural estimates of auditory thresholds in hens

Six hens were trained in a trial-by-trial auditory discrimination in which trials were signalled by the lighting of two key lights. Correct responses (left-key pecks when a tone was presented and right-key pecks when it was not) produced intermittent deliveries of food. Following training at one tone frequency and intensity, testing sessions at lower intensities were carried out. Training and testing were repeated for 10 frequencies between 260 Hz and 8000 Hz. Derived measures of detection performance allowed estimates of the intensities at which no discrimination would have occurred. The threshold measures are presented and suggest that the range of most sensitive hearing was between 3000 and 5000 Hz.     

Experimental exposure of llamas (Lama glama) to Brucella abortus: humoral antibody response

Positive antibody reactions to brucella were observed in the sera of four llamas receiving Brucella abortus Strain 19 subcutaneously at 2-3 weeks post-exposure (PE) using five of eight conventional brucella serologic tests and an ISU-ELISA. Positive brucella antibody reactions were detected in sera of four llamas exposed by intraocular instillation (IOI) of 1.02x10(8) (high dose) B. abortus Strain 2308 at 16-35 days PE using seven of eight serologic tests or an ISU-ELISA. Brucella antibody was also detected in sera of four llamas exposed by IOI of 9x10(5) (low dose) B. abortus using each of four agglutination tests, Complement Fixation test, PCFIA, the rivanol test and the ISU-ELISA at 16-35 days PE. Positive reactions were observed using the Card test, BAPA, SPT, STT, the rivanol test, the PCFIA, and the ISU-ELISA on sera collected on days 42-70 PE, except on one llama, given the low dose; that llama was negative on the PCFIA on day 42. Positive or suspicious reactions were not detected in sera of controls, receiving saline subcutaneously, using the routine tests, with the exception of the CFT. The B. abortus Strain 2308 was isolated from tissues of seven of eight llamas exposed to virulent B. abortus Strain 2308.     

Return-to-sensibility problems after penetrating captive bolt stunning of cattle in commercial beef slaughter plants

OBJECTIVE: To evaluate efficacy of penetrating captive bolt stunning of cattle in commercial beef slaughter plants and identify potential causes of a return to sensibility among stunned cattle. DESIGN: Observational study. SAMPLE POPULATION: 21 federally inspected commercial beef slaughter plants. PROCEDURE: In each plant, stunning of at least 100 cattle (19 large plants) or a minimum of 1 hour of production (2 small plants) was observed, and cattle were evaluated for signs of returning to sensibility on the bleed rail. Cattle with a limp, flaccid head, a lack of spontaneous blinking, and an absence of a righting reflex were considered insensible. RESULTS: In 17 of the 21 (81%) plants, all cattle were rendered insensible before they were hoisted onto the bleed rail. The remaining 4 plants had cattle that had signs of returning to sensibility; these cattle were restunned prior to skinning or leg removal. Of 1,826 fed steers and heifers, 3 (0.16%) had signs of returning to sensibility, whereas 8 of 692 (1.2%) bulls and cows did. Return-to-sensibility problems were attributed to storage of stunner cartridges in damp locations, poor maintenance of firing pins, inexperience of the stunner operator (ie, shooting cattle too high on the forehead), misfiring of the stunner because of a dirty trigger, and stunning of cattle with thick, heavy skulls. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that efficiency of captive bolt stunning of cattle in commercial slaughter plants can be safely and objectively assessed. Care should be taken to maintain stunners correctly, particularly when stunning bulls and cows with heavy skulls.