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.     

Amino acid digestibilities in different samples of wheat shorts for growing pigs.

An experiment was carried out to investigate the variability in apparent ileal amino acid (AA) digestibilities in simulated samples of wheat shorts consisting of different proportions of wheat bran (WB), wheat shorts (WS), and wheat flour (WF), hereafter referred to as wheat fractions. The proportions of WS, WB, and WF and the NDF content (DM basis) of the wheat fractions were as follows: A, 70% WS, 30% WB, and 42.3% NDF; B, 85% WS, 15% WB, and 41.8% NDF; C, 100% WS and 41.3% NDF; D, 85% WS, 15% WF, and 35.2% NDF; and E, 70% WS, 30% WF, and 29.5% NDF. Six barrows, average initial BW of 37.2 kg, fitted with a simple T-cannula at the distal ileum, were fed one of six experimental diets according to a 6 x 6 Latin square design. Six diets were formulated to contain 17% CP (as-fed basis). Diets A, B, C, D, and E contained 17.53% soybean meal (SBM), which contributed 50% of the CP in these diets. The wheat fractions contributed the remaining 50% of the CP in these diets. Diet F contained 35.05% SBM, which was the sole source of dietary CP. Chromic oxide was used as the digestibility marker. During the first experimental period, the daily dietary allowance was provided at a rate of 5% (wt/wt) of the average BW. Thereafter, the daily dietary allowance was increased by 100 g at each successive period. Each experimental period comprised 12 d. Following a 7-d adaptation period, feces were collected for 48 h and ileal digesta for a total of 24 h. The apparent ileal digestibilities of AA in the wheat fractions were calculated using the difference method. The digestibilities were usually lowest in the wheat fractions containing WB and highest in those containing no WB. The average of the digestibilities of the indispensable AA was 63.5% for wheat fraction A, which contained 30% WB, and 71.9% for wheat fraction C, which did not contain WB. There were no differences (P > .05) in lysine digestibilities among the wheat fractions, which ranged from 54.7 to 64.1%. Of the indispensable AA, with the exception of arginine, lysine, and methionine, the apparent ileal digestibility values of AA were negatively correlated (P < .05) with the NDF content in the wheat fractions.     

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.     

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.     

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.     

Evaluation of iodophor skin preparation techniques and factors influencing drainage from ventral midline incisions in horses.

OBJECTIVE: To document natural bacterial flora on the ventral aspect of the equine abdomen, to compare 2 preparation techniques, and to identify potential risk factors that may contribute to incisional drainage. DESIGN: Prospective study. ANIMALS: 53 horses undergoing exploratory celiotomy. PROCEDURE: Group-1 horses (n = 26) were prepared with povidone-iodine and alcohol. Group-2 horses (27) were prepared with a film-forming iodophor complex. Numbers of bacterial colony-forming units (CFU) were measured before and after surgical scrub, following skin closure, and after recovery from general anesthesia. Swab specimens to identify normal skin bacterial flora and potential pathogens were obtained by swabbing a 4 x 4-cm area. Variables that might affect incisional drainage were also investigated. RESULTS: For both techniques, there was a significant reduction in bacterial numbers after skin preparation. Incisional drainage was observed in 14 (26%) horses (8 group-1 and 6 group-2 horses). Preexisting dermatitis, poor intraoperative drape adherence, high number of bacterial CFU obtained after recovery from anesthesia, and high number of CFU obtained from the surgery room environment were the main risk factors associated with subsequent incisional drainage. Bacillus spp, nonhemolytic Staphylococcus spp, Micrococcus spp, Corynebacterium spp, Streptomyces spp, other nonenteric genera, and nonhemolytic Streptococcus spp were the most common isolates obtained before surgical scrub. CONCLUSIONS AND CLINICAL RELEVANCE: Both skin preparation techniques were equally effective in reducing numbers of bacterial CFU by 99%, and a significant difference was not found in incisional drainage rate between groups. Protection of the wound during recovery from anesthesia and the immediate postoperative period may reduce incisional drainage after abdominal surgery in horses.     

Binding of bovine IgG2a and IgG2b allotypes to protein A, protein G, and Haemophilus somnus IgBPs.

Immunoglobulin binding proteins (IgBPs) are thought to be virulence factors which enable pathogens to evade the host's immune response. Since bovine IgG2 is important in protection against pyogenic infections, the binding characteristics of Staphylococcus aureus protein A (PrA), streptococcal protein G (PrG), or Haemophilus somnus high molecular weight IgBPs to the two bovine IgG2 allotypes were examined. For PrA or PrG binding of IgG2, guinea pig red blood cells coated with specific IgG2a or IgG2b antibodies were used in a competitive binding inhibition assay with unlabeled and horseradish peroxidase-labeled PrA or PrG. To determine which sizes of H. somnus. IgBPs bind to the two IgG2 allotypes, immunoblots with H. somnus culture supernatant were probed with anti-DNP IgG2a and IgG2b. This detects only Fc binding because anti-DNP does not cross-react with H. somnus antigens. Both IgG2 allotypes bound equally well to PrA and PrG. However, IgG2b but not IgG2a bound to H. somnus high molecular weight IgBPs. The lack of differential binding of bovine IgG2 allotypes to PrA and PrG means that these IgBPs can be considered to be unbiased reagents in assays for detection of bovine IgG2 or for immunoaffinity purification. The differential binding of H. somnus IgBPs to the IgG2 allotypes indicates that animals having one allotype may be more resistant to H. somnus infection than animals having the other allotype.     

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.     

Serum total and ionized magnesium concentrations and urinary fractional excretion of magnesium in cats with diabetes mellitus and diabetic ketoacidosis

OBJECTIVE: To determine magnesium (Mg) status in cats with naturally acquired diabetes mellitus (DM) and diabetic ketoacidosis (DKA), evaluate changes in Mg status after treatment for DKA, and correlate Mg status with systemic blood pressure and degree of glycemic control. DESIGN: Case series and cohort study. ANIMALS: 12 healthy cats (controls), 21 cats with DM, and 7 cats with DKA. PROCEDURE: Serum total magnesium (tMg) and ionized magnesium (iMg) concentrations and spot urinary fractional excretion of magnesium (FEmg) were determined, using serum and urine samples obtained from all cats when they were entered in the study and from cats with DKA 12, 24, and 48 hours after initiating treatment. Indirect blood pressure and degree of glycemic control were determined in 10 and 21 cats with DM, respectively. RESULTS: Initially, 2 and 13 cats with DM and 1 and 4 cats with DKA had serum tMg and iMg concentrations, respectively, less than the low reference limit (mean-2 SD) determined for controls. In cats with DKA, serum tMg concentration decreased significantly over time after initiating treatment. Urinary FEmg was significantly higher in cats with DM or DKA, compared with controls. Systemic hypertension was not detected nor was there a correlation between Mg status and degree of glycemic control in cats with DM. CONCLUSIONS AND CLINICAL RELEVANCE: Hypomagnesemia was a common finding in cats with DM and DKA and was more readily identified by measuring serum iMg concentration than tMg concentration. The clinical ramifications of hypomagnesemia in such cats remain to be determined.