Quantitative Determination of Proteolytic,Lipolytic and Haemolytic Activities in Semen of Some Animal Species

1. Methods for quantitative determination of proteolytic, lipolytic and haemolytic activities and egg yolk factors in semen are described.2. Comparative quantitative determinations of these enzyme activities in the semen of goat, ram, bull, boar, dog and horse have been carried out.3. Special interest attaches to the egg yolk turbidity factor which is found in goat semen and, in small concentrations, also in boar semen.4. As each species appears to have its specific enzyme pattern, regular enzymatic control does not seem to have great significance.     

Proteases of Clostridium botulinum. V. Studies on the serological relationship between proteases from Clostridium botulinum and other spore-forming bacteria

By the use of the electrophoretic casein precipitating inhibition test (CPI-test) the serological relationship between proteolytic enzymes produced by different species within the genera Clostridium and Bacillus has been tested. The proteases produced by Clostridium botulinum types A, B, C, D and F cross-reacted with each other. Clostridium botulinum strain 84 was inhibited by antiproteases produced against Clostridium sporogenes, Clostridium botulinum types C and F (protease F I and F II), but not by antiproteases against Clostridium botulinum types B and F (protease II), Clostridium bifermentans and Clostridium perfringens. The protease of the newly described Clostridium botulinum strain 89 (type G) was inhibited by Clostridium sporogenes antiprotease, but not by any of the other antiproteases. It is not possible to differentiate between Clostridium botulinum, Clostridium sporogenes and Clostridium perfringens by use of serological differentiation of their proteolytic enzymes. The protease of Clostridium bifermentans is not serologically related to any of the species tested in this investigation. Proteases produced by different Bacilli were not inhibited by antiproteases from Clostridium botulinum types B, C and F, Clostridium sporogenes, Clostridium bifermentans, and the two strains of Clostridium perfringens tested. This investigation indicates a serological relationship between proteases from different Clostridium species, but not a serological relationship between proteases produced by the Clostridium species and Bacillus species tested.     

Transvenous retrograde portography for identification and characterization of portosystemic shunts in dogs

Transvenous retrograde portography for identification and characterization of portosystemic shunts in dogs A method for transvenous retrograde portography (TRP) in dogs suspected to have a portosystemic shunt (PSS) and results in 20 dogs are described. For TRP, dogs were anesthetized and positioned in left lateral recumbency A dual-lumen balloon-tipped catheter was inserted into the right jugular vein and advanced into the azygos vein. The balloon was inflated to occlude the azygos vein, and contrast material was injected during fluoroscopic evaluation. The catheter was then positioned in the caudal vena cava just cranial to the diaphragm. The balloon was again inflated to occlude the vena cava, and contrast material was again injected. Once a shunt was identified, selective catheterization was attempted with a guide wire and angled catheter. A PSS was identified in 18 of the 20 dogs. In 10 of the 18, the shunt vessel could be selectively catheterized, allowing measurement of portal pressures while the shunt was occluded with the balloon. In 1 dog, results of TRP were normal, but subsequent exploratory celiotomy revealed a single extrahepatic PSS, which was surgically attenuated. The other dog in which results of TRP were normal did not have a macroscopic PSS. In dogs suspected to have a PSS, TRP may be a useful adjunctive diagnostic test that is less invasive than operative mesenteric vein portography and allows measurement of portal pressures before and after temporary shunt occlusion.     

Suppressor activity of bovine Fc gamma positive cells during a persistent infection with Mycobacterium avium

The cell mediated immune response (CMI) was measured in calves after experimental infection with Mycobacterium avium. Using the tuberculin skin test a CMI response could be measured from four to 14 weeks after infection, and with a lymphocyte stimulation (LS) test from six to 40 weeks. One year after infection no CMI response was detected by either of the tests, in spite of the fact that in such calves M avium bacteria could be found in the intestinal lymph nodes at autopsy. After removal of mononuclear cells bearing receptors for the Fc part of IgG, the peripheral blood lymphocytes obtained from a calf infected one year earlier responded to M avium pure protein derivative in the LS test in contrast to lymphocytes obtained from uninfected calves.     

Melatonin Supplementation During In Vitro Maturation and Development Supports the Development of Porcine Embryos

Melatonin has been reported to improve the in vitro development of embryos in some species. This study was conducted to investigate the effect of melatonin supplementation during in vitro maturation (IVM) and development culture on the development and quality of porcine embryos. In the first experiment, when the in vitro fertilized embryos were cultured with different concentrations of melatonin (0, 10, 25 and 50 ng/ml) for 8 days, the blastocyst formation rate of embryos cultured with 25 ng/ml melatonin (10.7%) was significantly increased (p < 0.05) compared to the control embryos cultured without melatonin (4.2%). The proportion of DNA‐fragmented nuclei in blastocysts derived from embryos cultured with 50 ng/ml melatonin was significantly lower (p < 0.05) than that of embryos cultured without melatonin (2.1% vs 7.2%). In the second experiment, when oocytes were cultured in the maturation medium supplemented with different concentrations of melatonin (0, 10, 25 and 50 ng/ml), fertilized and then cultured with 25 ng/ml melatonin for 8 days, there were no significant differences in the rates of cleavage and blastocyst formation among the groups. However, the proportions (2.7–5.4%) of DNA‐fragmented nuclei in blastocysts derived from oocytes matured with melatonin were significantly decreased (p < 0.05) compared to those (8.9%) from oocytes matured without melatonin, irrespective of the concentration of melatonin. Our results suggest that supplementation of the culture media with melatonin (25 ng/ml) during IVM and development has beneficial effects on the developmental competence and quality of porcine embryos.     

Low-Temperature Nonoxidative Activation of Methane over H-Galloaluminosilicate (MFI) Zeolite

Conversion of methane to higher hydrocarbons by its low-temperature activation without forming undesirable carbon oxides is of great scientific and practical importance. Methane can be highly activated, yielding high rates of conversion to higher hydrocarbons and aromatics (10 to 45 percent) at low temperatures (400° to 600°C), by its reaction over H-galloaluminosilicate ZSM-5 type (MFI) zeolite in the presence of alkenes or higher alkanes. The methane activation results from its hydrogen-transfer reaction with alkenes.     

Primary bovine viral diarrhoea virus infection in calves following direct contact with a persistently viraemic calf.

Six calves, aged 24 to 58 days and not previously exposed to bovine viral diarrhoea virus (BVDV), were infected with this agent by nose-to-nose contact with a persistently BVDV viraemic calf. The study was conducted in two trials, using 3 calves in each. All 6 calves showed a peak interferon level in serum at 4 days post infection (dpi), and they seroconverted to BVDV at 16-21 dpi. The calves in trial 1 had diarrhoea for 2 or 3 days between 2 and 6 dpi and one calf again from 9 to 11 dpi. During the periods of fever, the calves were slightly depressed. Those in trial 2 were more depressed and their oral and nasal mucous membranes were reddened but they never had diarrhoea. In both trials, fever (up to 41.3 degrees C) was a prominent symptom at 8 to 9 dpi and 2 calves showed a diphasic fever course. Respiratory affection was mild and no medical treatment was required. Haematological assessment demonstrated a transient but significant leukopenia and lymphopenia at 4 dpi (P less than 0.01 and P less than 0.05 respectively) and 11 dpi (P less than 0.05 and P less than 0.01 respectively). A significant decrease in thrombocyte count was seen at 4 dpi (P less than 0.05, n = 3). This study has demonstrated that nose-to-nose contact is an effective way of transmitting BVDV from persistently infected to susceptible cattle.     

Detection of porcine MHC class II antigens in different immunoassays

A monoclonal antibody (Mab), Mab 4-24-11, to human class II major histocompatibility antigens has been tested in commonly used immunoassays for detection of porcine class II major histocompatibility antigens (SLA-D). In a radioimmunoprecipitation assay, Mab 4-24-11 reacted with proteins from mitogen-stimulated porcine mononuclear cells (MNC). The molecular weights of the precipitated proteins were approximately 34 and 28 Kd. In indirect immunofluorescence, approximately 25% of porcine MNC in suspension reacted with Mab 4-24-11. This percentage diminished to 14% when Ig bearing MNC were removed, while it increased to 36% when adherent MNC were enriched. Thus, it was concluded that Mab 4-24-11 cross-reacts with SLA-D. In a immunohistochemical study, Mab 4-24-11 reacted with cells in acetone fixed cryostat sections from the gastric mucosa and endometrium. These properties of Mab 4-24-11 make it useful as a tool for further studies on the porcine immune system.