Ultrastructure of porcine circovirus in persistently infected PK-15 cells.

The ultrastructure of porcine circovirus was examined in persistently infected porcine kidney (PK)-15 cells. Virus-infected PK-15 cells had large numbers of intracytoplasmic inclusions, and a few cells had intranuclear inclusions. Intracytoplasmic inclusions were dispersed throughout the cytoplasm but were most numerous in the perinuclear cytoplasm. Inclusion were of various sizes, round to oval, and electron dense and were of two general types. Inclusions of the first type were small (0.1-0.5 microm diameter), not surrounded by trilaminar membranes, and granular with indistinct margins that blended with surrounding cytoplasm. Some contained 12+/-2-nm-diameter icosahedral virions in loose aggregates or rarely forming paracrystalline arrays. Small inclusions could be sites of viral assembly or maturation. Intracytoplasmic inclusions of the second type were larger (0.5-5.0 microm diameter) and more numerous and had abrupt margins surrounded by trilaminar membranes. They were more electron dense than small inclusions and were heterogeneous, containing various proportions of aggregated virions, electron-dense crystalline lamellae of 5 nm periodicity, and/or whorls of myelinoid membranes. Virions usually formed paracrystalline arrays and occasionally were loosely aggregated. Larger inclusions were typical of autophagolysosomes. Intranuclear inclusions were not membrane bound and were often associated with reticulated nucleoli or aggregates of heterochromatin. Some inclusions were irregularly shaped aggregates of indistinct, circular 10-12-nm-diameter viruslike particles. Others were 0.1-1.0 microm in diameter, round or ring shaped, dense, and finely granular, with sharply demarcated margins.     

Experimental infection of vaccinated slaughter ostriches in a natural, open-air feedlot facility with virulent Newcastle disease virus.

The presence of virulent Newcastle disease virus (NDV) since the 1993-94 epidemic in southern Africa holds major implications for the export of ostrich products from this region. A challenge experiment with this field strain was conducted in open-air feedlot facilities under strict biosecurity measures. The experiment was designed to follow vaccination and preslaughter quarantine regulations currently enforced in South African export ostrich facilities in order to determine the viremia period and immune response under these specific circumstances. One hundred forty-three slaughter ostriches were allocated into three test groups, according to the time period between pretrial vaccination and challenge (1-2 mo, 2-4 mo, 4-6 mo), and an unchallenged control group. All birds in the test groups were challenged by oral, tracheal, and ocular routes with a field isolate of NDV. They were slaughtered over the next 4 wk on nine separate occasions and bled on 12 occasions. Virus isolation was attempted from seven sets of pooled samples from each bird to determine the viremia period and the serum antibody concentrations were measured by hemagglutination inhibition (HI) and enzyme-linked immunosorbent assay (ELISA) methods to establish an immune response curve. NDV could be back-isolated only up to day 9 postinfection and from only six ostriches with poor immune response titers and corresponding to a rise in antibody levels above an indirect ELISA optical density reading of 0.33. Virus could be recovered only from brain and respiratory tract tissue. The HI test was less sensitive than the ELISA. Immune response curves did not differ significantly between the groups and peaked on day 14 post-infection. From these data, ELISA titers would appear to be a good indicator of the probability that an ostrich will be clinically infected after velogenic NDV challenge. These results also suggest that the current vaccination schedule enforced by the South African Veterinary Authorities results in protective immunity in up to 95% of slaughter ostriches from export approved facilities. The standard 30-day preslaughter quarantine period introduced as part of Crimean-Congo hemorrhagic fever virus control measures also appears sufficient to encompass the determined NDV viremia period of 9-11 days in slaughter ostriches.     

Survival of Salmonella in the crop contents of market-age broilers during feed withdrawal.

Recent studies have indicated that crop contamination increases during preslaughter feed withdrawal and that contaminated crop contents may serve as an important source of Salmonella entry into poultry processing plants. During the present study, we evaluated the effect of preslaughter feed withdrawal on crop pH and Salmonella crop contamination in broilers from three commercial broiler flocks. The effect of experimental feed withdrawal on crop pH, lactic acid concentration, and Salmonella crop contamination was also evaluated in market-age broilers challenged experimentally with Salmonella typhimurium. Crop pH increased significantly (P < 0.05) from 3.64 +/- 0.25 before feed removal to 5.14 +/- 0.72 after 8 hr of feed withdrawal in broilers from commercial flocks. The incidence of Salmonella crop contamination in the commercial broilers increased (P < 0.05) from 3.3% before feed removal to 12.6% after 8 hr of feed withdrawal. Similarly, crop pH increased (P < 0.05) by a magnitude of approximately 1 unit in broilers after 8 hr of experimental feed withdrawal. The population of S. typhimurium in the crops of the experimentally challenged broilers increased (P < 0.05) by approximately 1 log unit during the 8-hr experimental feed withdrawal. The concentration of lactic acid in the crop of the broilers during experimental feed withdrawal decreased (P < 0.01) from a range of 119-135 mumol/ml before feed removal to a range of 22-32 mumol/ml after 8 hr of feed withdrawal. The results indicated that feed withdrawal resulted in a decrease in lactic acid in the crop, accompanied by an increase in crop pH, and an increase in Salmonella crop contamination.     

Determination of Normal Values Using an Automated Coagulation Timer for Activated Coagulation Time and its Application in Dogs with Hemophilia

The present study was performed to determine normal values for the Medtronic HemoTec automated activated coagulation time (ACT) analyzer (Medtronic HemoTec Inc, Parker, CO, distributed in Switzerland by Convergenza AG, Vaduz, Liechtenstein), and to evaluate its ability to detect dogs with hemophilia. ACT was measured in 43 healthy dogs presented to the Companion Animal Hospital, University of Bern, Bern, Switzerland, with the Medtronic HemoTec ACT analyzer to determine normal values. The mean +/- 2 standard deviations (SDs) of the values obtained was defined as the normal range. ACT was measured 8-10 times on the same day in 6 dogs to determine repeatability. ACT also was measured in 11 dogs with hemophilia and compared with a conventional visual ACT measurement test and with the activated partial thromboplastin time (APTT). ACT values of the 43 dogs used to determine normal values ranged from 66.5 to 97.0 seconds (mean, 79.3 seconds; SD, 7.35 seconds; median, 78.5 seconds). A range of 64-95 seconds (mean +/- 2 SDs) was defined as the normal range for the tested device. Repeatability was poor (r = 0.256). ACT values measured with the automated device did not correlate with ACT values measured with a conventional visual test or with APTT Sensitivity of the test was 90.9%, specificity was 98.0%, and accuracy was 96.7%. Variability in the test results was large and may lead to incorrect results. The automated measurement device was not superior to the conventional visual method in evaluating dogs with hemophilia.     

Development of a polymerase chain reaction to detect Vietnamese isolates of duck virus enteritis.

A polymerase chain reaction (PCR) method for the detection of duck virus enteritis (DVE) virus in tissues of infected and affected ducks, and in cell culture was developed. This required us to obtain specific nucleotide sequence information as we could not find any specific data about the genome of the virus. We found the assay to be highly effective in detecting the virus under experimental conditions and to be easily transferred to laboratories in Vietnam where it is being used in studies on the epidemiology of the disease. We have applied this simple and rapid diagnostic method to the detection of DVE isolates grown in cell culture and tissues from infected birds. The assay was also able to differentiate DVE from other avian herpesviruses, such as Marek's disease, infectious laryngotracheitis virus and goose herpesvirus.     

Assessment of the respiratory actions of intramuscular morphine in conscious dogs.

The actions on the respiratory system of 0.25, 0.5 and 1.0 mg kg(-1) morphine given intramuscularly were studied in conscious dogs. Dogs breathed oxygen with 0, 2 and 4 per cent CO(2), in that order, through a mask attached to a flow sensor and connected to a respiratory mechanics monitor. When a steady state period of respiration was reached breathing pure oxygen, respiratory rate, tidal volume, respiratory minute volume, peak expiratory flow rate and end tidal CO(2)(PetCO(2)) were measured. The respiratory minute volume and PetCO(2) were measured when the dogs breathed 2 and 4 per cent CO(2) in oxygen, the points plotted onto a graph and the gradient of the line, describing the PCO(2)/ventilation response, plus the intercept with the y-axis were determined. Measurements for each morphine dose were taken before injection and at 30 minutes, 1, 2, 3, 4, 6 and 8 hours post injection.The incidence of panting after morphine was dose related and it occurred in all dogs given the high dose. Morphine reduced the gradients of the PCO(2)/ventilation response lines and raised the intercept. Other changes were increased respiratory minute volume and peak expiratory flow and decreased PetCO(2) and tidal volume.     

Interbreed variation of DLA-DRB1, DQA1 alleles and haplotypes in the dog.

Although 36 DLA-DRB1 and 10 DLA-DQA1 allele sequences have been published to date, no data on individual allele frequencies exists, either for specific breeds or cross breeds, and the full extent of the polymorphism at each of these loci is still not known. We have used sequence-specific oligonucleotide probing (SSOP) to characterise a series of 367 dogs for their DRB1 and DQA1 alleles. These included individual animals from over 60 different breeds, with numbers per breed ranging from 1 to 39. DLA types were generated from 218 dogs for DRB1 and from 330 dogs for DQA1, while 181 dogs were characterised for both these loci. The frequency of individual DRB1 and DQA1 alleles showed considerable interbreed variation, e.g. 83% of West Highland White Terriers were DRB1*01 as opposed to 9% of Collies. No breed had >9 of the 22 DRB1 types defined in this study; several breeds had only two DRB1 types. DLA-DQA1 showed less variation in allele numbers per breed, but also showed considerable interbreed frequency variation. Haplotype analysis revealed over 44 different DRB1/DQA1 combinations. Of these, 25 were in a number of animals, and also in an animal that was homozygous for one or both of these loci. Some DRB1 alleles could be found in combination with several different DQA1 alleles, while others were only present in one haplotypic combination. DLA allele frequency data in normal dogs will be critical for disease association studies. It may also be possible to use haplotype data to establish the genetic relationships between different dog breeds.     

Influence of body condition score on live and carcass value of cull beef cows.

Mature beef cows (n = 88) were slaughtered to determine the influence of body condition score (BCS) on carcass and live animal value. Cows were weighed and assigned a BCS (9-point scale), 24 h before slaughter. Hide and by-products weights were recorded during harvest. After a 48-h chill period, the right side of each carcass was fabricated into boneless subprimal cuts, minor cuts, lean trim, fat, and bone. Weights were recorded at all stages of fabrication. Carcass values (U.S.$/100 kg of hot carcass weight) were calculated for U.S. Utility and U.S. Cutter grades, as well as for the Utility/Cutter mix for each BCS. Gross value included the carcass value and the value of the hide and byproducts, whereas net value was calculated after harvest and fabrication costs and by-product value were considered. Live value (U.S.$/100 kg of live weight) was computed by dividing the net value by the animal's live weight 24 h before harvest. The value of the hide and by-products for BCS-2 cows was greater (P<.05) than for cows assigned a BCS of 3 through 8. Even though U.S. Utility carcasses from BCS-8 cows produced the least (P<.05) valuable subprimal cuts from the chuck, loin, and round, the gross and net values of BCS-8 cows were greater (P<.05) than those of BCS-3, 4, 5, and 6. Within the grade of U.S. Cutter, carcasses from BCS-6 cows had the highest (P<.05), and BCS-2 cows had the lowest (P<.05), gross and net values. Across the U.S. Utility/Cutter mix, cows designated with a BCS of 7 and 8 had greater (P<.05) gross and net values than cows assigned a BCS of 6, or lower. Live value increased linearly (P = .0002) from a low of $76.10/100 kg for BCS-2 cows to a high of $90.84/100 kg for BCS-7 cows. Carcasses from BCS-6 cows were relatively lean (8.4 mm of fat opposite of the longissimus muscle), and approximately 73% of the carcasses achieved a quality grade of U.S. Utility. Moreover, carcasses from BCS-6 cows had the highest total carcass values and live values comparable (P>.05) to BCS-7 cows. Information from this study can be used by the non-fed beef industry to establish a value-based marketing system. Data from this study would indicate that marketing cull beef cows at a BCS of 6 could optimize economic returns to both cow-calf producers and non-fed beef packers.     

Estimation of Neospora caninum seroprevalence in dairy cattle from Normandy, France.

An epidemiological study was conducted in Orne (France) on randomly selected dairy herds (42 herds including 1,924 cows and heifers, which were at least 15 months old). The aim was primarily to estimate the seroprevalence of Neospora caninum infection from two blood samples per cow, using an enzyme-linked immunosorbent assay (ELISA) for N. caninum (one positive result indicating infection). The second aim was to test the association between some individual and herd factors and N. caninum seropositivity with a logistic model including a random term effect. The prevalence was estimated at 5.6% (107 seropositive animals). At least 27 of the 42 herds had one seropositive cow or heifer. The intra-herd seroprevalence varied from 1.1 to 8% for 18 positive herds (66.7%). Dogs were present in 36 farms and 104 of the 107 seropositive animals were exposed to them. The factors associated with individual seropositivity were the presence of cats (OR = 0.17; P < 0.001), dogs (OR = 4.35; P = 0.02), rabbits and/or ducks (OR = 2.10; P = 0.04), long calving periods (12 months) (OR = 0.44; P = 0.007), tethered housing (OR = 2.50; P = 0.01), somatic cell counts (200-400 x 10(3) cells/mL) (OR = 0.24; P < 0.001) and pond water supply (OR = 2.43; P = 0.04). In conclusion, the animal and intra-herd seroprevalences were low in dairy cows from Normandy, France.     

Effects of hemolysis on plasma electrolyte and chemistry values in the common green iguana (Iguana iguana).

Hemolysis of serum and plasma samples is a common problem in veterinary diagnostic laboratories. We measured the effects of hemolysis on nine plasma analytes in 10 clinically normal common green iguanas (Iguana iguana). Blood samples with moderate and marked hemolysis were produced from each iguana by freezing, centrifuging, and decanting plasma from a portion of each blood sample, and combining the nonhemolyzed plasma with different amounts of hemolyzed plasma from the same individual. Moderate hemolysis significantly increased plasma phosphorus levels. Marked hemolysis significantly increased plasma values of potassium, phosphorus, total protein, and aspartate aminotransferase. The severity of hemolysis must be considered when interpreting values for these analytes in iguana plasma.