Distribution of ASFV antigens in pig tissues experimentally infected with two different Spanish virus isolates.

Lymph nodes, spleen, liver, lung and kidney obtained from pigs experimentally infected with two African Swine Fever Virus (ASFV) isolates of differing virulence were fixed by perfusion with glutaraldehyde and embedded in paraffin. An immunoperoxidase technique using a polyclonal anti-ASFV serum was performed on tissue sections in order to detect ASFV antigen. The distribution of ASFV antigen in such infected organs is shown and the differences between both infections compared and discussed. Monocytes, macrophages, hepatocytes, endothelial cells, neutrophils and epithelial cells were found to contain ASFV antigens.     

Pathology of infectious diarrhea of infant rats (IDIR) induced by an antigenically distinct rotavirus

Suckling rats were inoculated with a group B rotavirus to determine the progression of the morphologic changes induced in the intestine by this virus. Several changes were observed by light microscopy 1 day after viral inoculation: shortening of small intestinal villi, villous epithelial necrosis, and villous epithelial syncytia. The lesions were most often present in the distal small intestine, although other small intestinal segments were affected to a lesser degree. By day 3 post-inoculation, epithelial necrosis, and syncytia were no longer present; however, the villous epithelium was disorganized and irregularly vacuolated, and intestinal crypt epithelium was hyperplastic. Alterations in villous height to crypt depth ratios were present in portions of the small intestine for the remainder of the 12-day study period. Epithelial syncytia appeared to form by the breakdown of the lateral interdigitating membranes of the absorptive villous epithelium. Viral particles, abundant in the syncytia, appeared to form from amorphous or reticular arrays of viral precursor material. Group B rotaviral antigens, as detected by indirect immunofluorescence, were present in large amounts in the small intestinal villous epithelium only on the first day after viral inoculation. These studies show that two important diagnostic features of group B rotaviral infections of rats, epithelial syncytia and viral antigen as determined by immunofluorescence, are present only on the first day of disease. These findings should be taken into consideration when attempting to diagnose disease induced by this agent.     

Insulin-like growth factor I receptor analysis of satellite cell-derived myotube membranes established from two lines of Targhee rams selected for growth rate

Ovine-derived fibroblasts were used to validate an insulin-like growth factor I (IGF-I) membrane-receptor binding assay system. Competitive binding using fibroblasts revealed that half-maximal inhibition of 125I-IGF-I binding by IGF-I was 2.3 nM. SDS-polyacrylamide gel electrophoresis analysis of specific protein-associated 125I-IGF-I was consistent with the migration of 125I-IGF-I-labeled Type I IGF receptor alpha-subunits at Mr 133,000 daltons. Further, the efficiency of two cell solubilization methods was examined and time-dependent binding equilibrium was determined for the membrane assay system. Satellite cell-derived myotubes were subsequently isolated from primary satellite cell cultures established from the semimembranosus muscles of high and low efficiency-of-gain (EOG) Targhee rams, and IGF-I receptor dynamics were measured. A membrane competitive binding study revealed that half-maximal inhibition of 125I-IGF-I binding was achieved by 1-ng IGF-I for low, and 10-ng IGF-I for high, EOG myotube membrane preparations. Kd values were similar between the high EOG (4.78 nM) and low EOG (2.95 nM) groups; however, receptor concentrations (Bmax) appeared to differ between groups. High EOG membrane receptor Bmax was 3.88 pmole/micrograms protein (19.87 pmole/micrograms DNA), whereas low EOG membrane receptor Bmax was 1.22 pmole/micrograms protein (9.28 pmole/micrograms DNA). These preliminary findings support the hypothesis that genetic selection for EOG results in altered satellite cell responsiveness to IGF-I.     

Correlation of DNA ploidy to tumor histologic grade, clinical variables, and survival in dogs with mast cell tumors.

By using flow cytometry, a retrospective analysis of the DNA content of 40 primary canine mast cell tumors and seven lymph nodes that contained metastatic mast cell tumor from 44 dogs of various breed, sex, and age was performed on formalin-fixed, paraffin-embedded samples of the tumors and nodes. These samples were chosen according to the following criteria: samples contained sufficient well-preserved tumor tissue in the paraffin block for processing, sufficient patient history data were available, clean and homogeneous cell suspensions were obtained after processing, and interpretable DNA histograms were produced on analysis. The ploidy data obtained were compared with the histopathologic grade, the anatomical site of occurrence, the clinical stage of the tumors, and the survival of the dogs. Over 70% (29/40) of the mast cell tumors were diploid. Three metastatic mast cell tumors in lymph nodes had the same ploidy status as their corresponding primary tumors. In five dogs, mast cell tumors from multiple sites in each dog displayed similar ploidy status. Of 26 dogs evaluated for survival times, 69% (18/26) had diploid tumors and 31% (8/26) had aneuploid tumors. When numbers of diploid versus aneuploid tumors were compared, no significant difference was found between any two grades, clinical stages, or anatomic sites. A significant difference (P = 0.02) was found, however, between aneuploid and diploid tumors when comparing Stage I and non-Stage I disease. The Kaplan-Meier survival plot indicated a tendency towards an increased survival within the first year in dogs with diploid versus aneuploid tumors (P = 0.06).     

Selective measurement of lipoprotein lipase and hepatic triglyceride lipase in heparinized plasma from horses.

Affinity chromatography on heparin sepharose was used to identify 2 lipolytic enzymes in heparinized plasma from horses. One enzyme was typical of hepatic triglyceride lipase (HTGL), because it was resistant to inactivation by high concentrations of NaCl, and it did not require the addition of serum for activity. The other enzyme was identified as lipoprotein lipase (LPL), because of its inactivation at NaCl concentrations in excess of 0.2M, and its dependency on addition of serum as a source of apolipoprotein C-II activator. The enzymes were purified by 347-(HTGL) and 442- (LPL) fold, with yields of 54 and 58%, respectively. The partially purified enzymes were used to design incubation conditions that gave optimal activities for each enzyme in vitro. A selective assay was then developed for direct measurement of LPL and HTGL activities in heparinized plasma from horses. Analysis of HTGL took advantage of the almost complete inactivation of LPL when serum cofactor was excluded from the assay at the NaCl concentration that gave optimal HTGL activity. Prior incubation of heparinized plasma with sodium dodecyl sulfate to inhibit HTGL was necessary for measurement of LPL, because HTGL retained 67% of its activity at the NaCl concentration required for optimal LPL activity. Activity of each enzyme was measured in heparinized plasma from 12 Shetland ponies. The mean activity +/- SD for LPL was 3.22 +/- 1.04 mumol of fatty acids/ml of heparinized plasma/h (mumol of FA/ml/h. The mean activity for HTGL was 4.9 +/- 1.56 mumol of FA/ml/h. The performance of the assay was assessed by replicate analysis of pools of each enzyme with high and low activities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Use of 64Copper Measurements to Diagnose Canine Copper Toxicosis

Inherited canine copper toxicosis is a serious problem in Bedlington terriers and West Highland White terriers, and may also be a problem in other less-studied breeds. Affected dogs become ill at midlife with progressive and ultimately fatal liver disease. Treatments for removal of copper and prevention of copper accumulation are available, but are most effective if begun before the dog becomes ill. Until recently diagnosis has not been available until the dog is 1 year of age, and then only by an invasive liver biopsy with determination of liver copper concentration. The authors studied the use of 64copper for early diagnosis of canine copper toxicosis. Two procedures were evaluated. The first involved measuring the concentration of 64copper in blood 24 hours after oral administration of the radioisotope. At this time, 64copper was associated primarily with ceruloplasmin secreted into the blood by the liver. This procedure is useful in the diagnosis of the human counterpart, Wilson's disease. However, the authors found it to be nondiscriminatory between affected and unaffected dogs. In contrast, the second procedure, which involved measuring 64copper excreted in stool during 48 hours after an intravenous dose of radioisotope, yielded results that differentiated most affected and unaffected dogs.     

Serum Progesterone Concentrations and Blood Neutrophil Alkaline Phosphatase Activity in Cows Following Normal and Abnormal Parturition

Serum progesterone concentrations and alkaline phosphatase (ALP) activity of blood neutrophils were determined in 3 groups of cows (n = 5 each) on days 1 and 2 and then at 3-day intervals up to 32 days post-partum. Group I cows had a normal delivery, Group II cows had dexamethasone-induced parturition and Group III cows were subjected to a caesarian section. All cows in Group III and 2 cows in Group II retained their fetal membranes. Mean serum progesterone concentrations declined the second day after calving (to < 0.67 ng/ml) and remained at low levels (< 0.54 ng/ml) throughout the observation period, except for the values in Group III, which were elevated on day 16 (0.94 ng/ml), declined again on day 26 (0.46 ng/ml) and peaked (1.05 ng/ml) on day 32 portpartum. Significant (P < 0.01) differences were found between serum progesterone concentrations on day 1 and on each of the other sampling days in Groups I and III. Day X parturition group interaction was significant (P < 0.05) for the progesterone concentrations. No significant differences were found between the overall means of ALP activity of blood neutrophils in the 3 parturition groups nor between days of the experiment. No significant correlation was found between serum progesterone concentrations and ALP activity values of blood PMN during the first 32 days post-partum. Inhalt: Serum Progesteron Konzentrat und Aktivität der alkalischen Phosphatase in neutrophilen Blutzellen bei Küken mit normalem und abnormalen Geburtsverlauf Serum-Progesteronkonzentrationen und alkalische Phosphatase- (ALP) Aktivität von neutrophilen Granulozysten, nach der Abkalbung am Tag 1 und 2 und dann im Abstand von 3 Tagen bis zum 32. Tag p.p. bei Kühen untersucht, die in drei Untersuchungsgruppen (n = 5) eingeteilt waren. Gruppe I kalbte normal ab, in Gruppe II wurde die Geburt durch Dexamethason eingeleitet, und Kälber der Gruppe III wurden via Kaiserschnitt gewonnen. Alle Tiere der Gruppe III und 2 Kühe der Gruppe II wiesen Nachgeburtsverhaltungen auf Der mittlere Serum-Progesterongehalt sank am 2. Tag p.p. auf < 0,67 ng/ml ab und blieb auf niederigem Niveau (< 0,54 ng/ml) während der gesamten Untersuchungs-periode. Lediglich in Gruppe III, mit erhöhten Werten (0,94 ng/ml) an Tag 16 p.p., sanken die Werte wieder an Tag 26 p.p. (0,46 ng/ml) und erreichten einen Maximalwert an Tag 32 p.p. mit 1,05 ng/ml. Signifikante (P 0,01) Unterschiede in der Serum-Progesteronkonrentration wurden zwischen Tag 1 p.p. und allen anderen Untersuchungstagen in den Gruppen I und III gefunden. Interaktionen zwischen den Gruppen für Tag X waren signifikant (P 0,05). Keine signifikanten Unterschiede wurden für den Gesamtmittelwert der ALP-Aktivität in den neutrophilen Granulorysten zwischen den Gruppen und im Vergleich der Untersuchungstage gefunden. Es wurden auch keine signifikanten Korrelationen zwischen den Serum-Progesteronkonzentrationen und der ALP-Aktivität der PMN während der ersten 32 Tage p.p. nachgewiesen.     

Oral fowlpox vaccination in chickens.

Chickens were given various fowlpox vaccines on food pellets--a commercial vaccine (strain M), and the same strain after a single passage on chorio-allantoic membrane or in chicken embryo fibroblasts. All three oral vaccines induced antibodies at levels similar to those induced by commercial strain M administered to the wingweb. The oral vaccine derived from chorio-allantoic membrane gave protection similar to that obtained with vaccine administered by the wingweb, but this required a thousandfold more virus.