Fractionation of cock spermatozoa

An account is given of the methodology for fractionation of cock spermatozoa into head and tail fractions by ultrasonication, followed by sucrose density gradient centrifugation. Quantitative estimates of DNA attested to 89.4% purity of the head fraction and low contamination of tails with heads. Recovery of protein and malic dehydrogenase (MDH) activity, following sperm fractionation, averaged 94.3% and 95.7%, respectively. Contamination of the head fraction with tails, as assessed by MDH assay, was only 4.65%, and the purity of the tail fraction was 91%. Intensive succinic dehydrogenase (SDH) activity was histochemically localised in the separated tail fraction and in the tail portion of intact spermatozoa. However, SDH activity was discernible neither in the head fraction nor in the head of intact spermatozoa.     

Effect of xylazine on cerebrospinal fluid pressure in conscious horses.

Lumbosacral CSF pressure was measured in 6 horses via a catheter inserted through the lumbosacral space. Heart rate, facial artery pressure, central venous pressure, and CSF pressure were measured before IV injection of a saline solution control, for 15 minutes after saline solution injection, and for 60 minutes after the IV injection of 1.1 mg of xylazine/kg of body weight. Arterial pH and blood gases were analyzed before saline solution injection, 15 minutes after saline solution injection, and at 15, 30, and 60 minutes after xylazine injection. Constant craniocervical posture was maintained during sedation. Lumbosacral CSF pressure was significantly decreased for 15 minutes after xylazine injection. Diastolic arterial pressure was significantly increased 4 minutes after xylazine administration and diastolic and mean arterial pressure were increased at 6 and 8 minutes after xylazine administration. Small increases in systolic arterial blood pressure and central venous pressure, and a small decrease in heart rate were observed. There were no significant differences in the arterial blood gas values. It was concluded that IV injection of xylazine causes a decrease in intracranial pressure in healthy conscious horses. The effects may be different in horses with neurologic disease or cerebral trauma.     

Antibody responses to avian encephalomyelitis virus vaccines when administered by different routes

Antibody responses to a commercial avian encephalomyelitis virus (AEV) vaccine administered by different routes were measured by an enzyme-linked immunosorbent assay (ELISA). Responses to single doses of vaccine administered by the ocular route to 10% of a flock were comparable with those obtained when all birds received a single dose in the drinking water. However, ocular vaccination of 5% of the flock resulted in significantly lower responses than those obtained when 10% were vaccinated. Maternal antibody was shown by the ELISA to persist in chickens from vaccinated flocks for up to 21 days after hatching. Day-old chickens with serum absorbances of < 0.3 at 492 nm, as determined by the ELISA, were shown to be susceptible to intracerebral challenge with the neurotropic Van Roekel strain of AEV.     

Plant protection problems caused by phytonematodes in Greece1

The major nematode pests of cultivated plants in Greece include root-knot nematodes Meloidogyne spp. (in almost all plants except citrus), cyst-forming nematodes Globodera rostochiensis, G. pallida (in potato), Heterodera avenae (in cereals), Ditylenchus dipsaci (in vegetables and ornamental plants) and Tylenchulus semipenetrans (in citrus). Based on observations made on samples examined at Benaki Phytopathological Institute, it was confirmed that serious infestation of new kiwi orchards with Meloidogyne spp. originated from seedlings infested at the nursery. Also the wide distribution of the nematode Tylenchulus semipenetrans in citrus orchards was mainly attributed to infested seedlings. The above observations led us, in 1985, to the decision to study the nematological fauna of nurseries in Greece.     

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.     

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)     

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.