Bovine Leukocyte Adhesion Deficiency in Danish Holstein-Friesian Cattle: I. PCR Screening and Allele Frequency Estimation

Jergensen, C. B., J. S. Agerholm, J. Pedersen and P. D. Thomsen: Bovine leukocyte adhesion deficiency in Danish Holstein-Friesian Cattle I. PCR screening and allele frequency estimation. Acta vet. Scand. 1993,34,231-236.–A screening program for bovine leukocyte adhesion deficiency (BLAD) in Danish Holstein-Friesian cattle has been initiated. During the first months 1611 animals were tested by a PCR based assay. Of these animals 1256, 346, and 8 were assigned normal, BLAD carriers, and BLAD affected animals, respectively One bull, born as a co-twin, showed weak reaction for the BLAD allele on DNA isolated from leukocytes, but a normal genotype on DNA isolated from semen. Chromosome analysis showed that this bull was a blood chimaera. Estimation of the BLAD allele frequency upon the PCR test results showed that around 450 Danish calves born in 1991 might have been affected with the recessive disorder.     

Cerebellar cortical abiotrophy in Wiltshire sheep

AIM: To investigate the nature of a neurological disease in Wiltshire sheep. METHODS: Three affected lambs were examined, humanely killed and necropsied. Selected neurological tissues were examined by light and electron microscopy. RESULTS: Primary neurological lesions were confined to the cerebellum and were characterised by loss of Purkinje cells and the presence of large hypertrophied dendrites of surviving Purkinje cells. These contained stacks of smooth endoplasmic reticulum. There was hyperplasia and cell swelling of Bergmann glia. Mild Wallerian-type degeneration affected white matter in the cerebellum and spinal cord. CONCLUSION: The cerebellar lesions were of a degenerative and reactive rather than hypoplastic nature. These, and the history, suggest a genetic cause with putative inheritance as an autosomal recessive trait. Accordingly, the disorder is described as a cerebellar abiotrophy.     

Outbreak of clinical listeriosis in sheep: evaluation from possible contamination routes from feed to raw produce and humans

We report the results of clinical and microbiological investigations on Listeria monocytogenes infections in a flock of 55 sheep and describe the implications for the safety of the raw milk and raw-milk cheeses produced in the on-farm dairy. The outbreak was caused by feeding grass silage, which was contaminated with 5 log10 CFU L. monocytogenes/g. Clinically, although having been fed from the same batch of silage, abortive (nine ewes), encephalitic (one ewe) and septicaemic (four ewes) forms of listeriosis were observed during the outbreak phase. As the starting point of feeding the contaminated silage was known we could calculate an incubation period of 18+/-2 and 26 days for the abortive and the encephalitic form of listeriosis, respectively. Pathologically, the septicaemic cases suffered from Listeria accumulation at comparable numbers in visceral organs but not in the brain. Only a single ewe developed central nervous symptoms and a rhomb-encephalitis was immunohistologically confirmed. In this case the infection proceeded from the nasal mucosa into the brain, with no infections of the liver, spleen and other visceral organs. Sampling of the cheese production chain, the farm environment and the persons living at the farm revealed the exposure of a farm-worker to an isolate genetically indistinguishable from the outbreak clone, obviously through the consumption of faecally contaminated bovine raw milk. The cheese under processing was free of Listeria because, as a result of intensive consultations, the farmer ensured a proper acidification of the cheese. The epidemiological findings suggest that food safety matters should be assessed in any case where infection of food-producing animals with potential human pathogens is observed.     

Construction and characterization of a fowlpox virus field isolate whose genome lacks reticuloendotheliosis provirus nucleotide sequences

Fowlpox virus (FWPV) has been isolated from vaccinated chicken flocks during subsequent fowlpox outbreaks that were characterized by a high degree of mortality and significant economic losses. This inability of current vaccines to induce adequate immunity in poultry could be reflective of an antigenic and/or biologic distinctiveness of FWPV field isolates. In this regard, whereas an infectious reticuloendotheliosis virus (REV) provirus is present in the majority of the field viruses' genomes, only remnants of REV long terminal repeats (LTR) have been retained in the DNAs of each vaccine strain. Although it has not been demonstrated whether the partial LTRs can provide an avenue for FWPV to reacquire the REV provirus by homologous recombination, utilizing viruses of which genomes lack any known integrated retroviral sequences could resolve concern over this issue. Therefore, such an entity was created by genetically modifying a recently isolated field strain of FWPV. This selection, in lieu of a commercial vaccine virus, as the progenitor was based on the probability that a virus circulating in the environment would be more antigenically similar to others in this locale and thus might be a better candidate for vaccine development. A comparison in vivo of the pathogenic traits of the parental wild-type field isolate, its genetically modified progeny, and a rescue mutant in whose genome the REV provirus was inserted at its previous location, indicated that elimination of the provirus sequence correlated with reduced virulence. However, even with elimination of the parasitic REV, the modified FWPV was still slightly more invasive than a commercial vaccine virus. Interestingly, both types of attenuated FWPV elicited a similar degree of antibody production in inoculated chickens and afforded them protection against a subsequent challenge by a field virus, the origin of which was temporally and geographically distinct from that of the progenitor strain. Due to its antigenicity being retained despite a decrease in virulence, this REV-less FWPV could potentially be developed as a vaccine against fowlpox.     

Seasonal Effect on Zebu Embryo Quality as Determined by Their Degree of Apoptosis and Resistance to Cryopreservation

In order to optimize the production of embryos under tropical conditions and to test a possible seasonal effect on embryo quality, 40 Zebu cows were superovulated during the dry season (April to May) and during the rainy season (July to August). A total of 116 (average 2.7/cow) and 83 embryos (3.5 average/cow) were obtained during the respective seasons. After classification as good, fair or poor quality, embryos were tested based on their ultrastructural differences (n = 53 dry season 16 good, 20 fair and 17 poor and n = 61 rainy season 21 good, 20 fair and 20 poor) and their degree of apoptosis using the TUNEL technique (n = 30 during the dry season and n = 55 in the rainy season). Structural characteristics determining embryo quality varied between good and fair quality embryos. No difference, however, was observed between good, fair and poor quality embryos from the two seasons. The number of TUNEL-positive cells was different among embryos (p < 0.001), being lower in labelled cells of good quality embryos regardless of the season. Fewer apoptotic cells were observed in embryos assigned in all three quality levels during the rainy season (p < 0.001). Ultrastructural evaluations confirmed the results obtained by TUNEL. Cryopreserved embryos of good (n = 25 in each season) and fair quality (n = 11 dry season; n = 17 rainy season) showed a significant decrease of TUNEL-positive cells during the rainy season (p < 0.05). Results suggest that embryos collected in the dry season have more cellular damage in contrast; embryos cryopreserved in the rainy season appeared morphologically better equipped to result in a pregnancy following transfer.     

Partial characterization of phagocytic activity in neutrophils of the nine-banded armadillo Dasypus novemcinctus

Though the armadillo is important as a research model in leprosy studies, the activity of armadillo's neutrophils is an aspect of little research. The aim of this study was carried out to partially characterize the chemotaxis, endocytosis and bacteriocidal ability of the neutrophils found in the nine-banded armadillo (Dasypus novemcinctus). Results showed that the chemotactic activity of the neutrophils, evaluated by the movement of the neutrophils through a nitrocellulose membrane (5 microm) in response to a chemo-attractive substance, was greater towards the armadillo serum (5.16+/-1.35 migration index, p<0.05) than towards the formil methionyl leucil phenylalanine (fMLP, 1.43+/-0.18 migration index) or human serum (0.56+/-0.18 migration index). Regarding endocytic capacity of the neutrophils and the monocytes against Escherichia coli was evaluated by a flow cytometry and using opsonized and non-opsonized E. coli-FITC at the following incubation times: 5, 10, 20, 30 and 60 min. The largest percentage of endocytosis by the neutrophils was 92.32+/-0.12% with opsonized bacteria and 77.73+/-14.33% with non-opsonized bacteria at 10 min incubation time, while the largest percentage of endocytosis by monocytes was 89.94+/-1.40% with opsonized bacteria and 73.07+/-15.6% with non-opsonized bacteria at 20 min incubation time. Evaluation of the bacteriocidal capacity of neutrophils using the methyl-thiazol-tetrazolium salts (MTT) reduction color-measurement assay showed an 89.0+/-10% mortality rate of non-opsonized E. coli and 89.0+10% of opsonized E. coli. In conclusion, the armadillo neutrophils show a good phagocytosis and bacteriocidal activity; however, a deficiency in the migration towards the fMLP was observed. This deficiency could be a cause so that the armadillo neutrophils do not respond quickly to invading microorganism.     

Q fever (Coxiella burnetii) investigations in dairy cattle: persistence of antibodies after vaccination.

The immune response and persistence of antibodies were investigated in dairy cattle vaccinated with formalin-inactivated phase (ph) I Coxiella burnetii vaccine agglutinating antibody geometric mean titer (GMT) of 193.2 at 1 month after vaccination compared to a GMT of 2.0 for nonvaccinated calves. The agglutinating antibodies gradually decreased in vaccinated cattle, but the GMT remained approximately 4 times higher than that for the nonvaccinated group for at least 20 months. Results of serotests at 2 months after revaccination indicated a rapid increase in the GMT to 177.0 with agglutinating titers between 1:64 and 1:512.     

Cardiopulmonary effects of fentanyl-droperidol, nitrous oxide, and atropine sulfate in dogs.

The cardiopulmonary effects of droperidol-fentanyl, nitrous oxide, and atropine were evaluated in 12 adult male Beagle dogs. All dogs were surgically instrumented with a cardiac output thermistor and arterial and venous catheters and were prepared with a chronic tracheostomy. Each dog was used as its own control, and data obtained when dogs were nonanesthetized and nonmedicated were compared with data recorded after the test drugs were administered. The dogs were randomly allotted to 3 groups of 4 dogs each. Group I dogs were given droperidol-fentanyl alone intravenously (IV); group II dogs were given droperidol-fentanyl IV with 67% nitrous oxide; and group III dogs were given atropine sulfate intramuscularly followed by droperidol-fentanyl IV with 67% nitrous oxide. Minute volume was decreased in the 3 groups of dogs for 3 to 5 minutes after droperidol-fentanyl was injected. This resulted in respiratory and metabolic acidosis in all dogs, as indicated by increased arterial carbon dioxide tension, decreased pH, and increased base deficit. In addition, droperidol-fentanyl given alone caused a decrease in systolic pressure and a slight decrease in heart rate. Group 1 dogs were sensitive to auditory stimulation. Cardiovascular changes were not seen when nitrous oxide was added; however, analgesia and muscle relaxation were improved. Premedication with atropine sulfate resulted in increased cardiac output, heart rate, and diastolic pressure, and subsequent administration of droperidol-fentanyl with nitrous oxide caused a transient increase in mean arterial and systolic pressure. This last anesthetic regimen, along with assisted or controlled respiration, seems to provide an excellent anesthetic state with minimal cardiopulmonary depression.