Versuche zur Zyklussynchronisation bei Kühen aus Beständen rnit Fertilitätsproblemen und bei Hybriden zwischen Wisent und Rind mit der PRID-Spirale*

Inhalt: An 81 Versuchs- und 92 Kontrolltieren der Rasse “Potnische Schwarzbunte” aus 3 Problembetrieben wurde ein Versuch zur Sterilitätsprophylaxe rnit der PRID-Spirale durchgeführt. Anhand klinischer Befunde und Milchprogesteronwerten wurde die Untergruppe aus Kühen mit Ovardystrophie (18 Versuchs- und 22 Kontrolltiere) gebildet und extra ausgewertet. Der Behandlungsbeginn lag zwischen 60 und 80 Tage post partum. Die Versuchstiere wurden mit der PRID-Spirale 12 Tage lang behandelt, die Kontrolltiere erhielten Injcktionen rnit Kochsalzlösung und solche rnit Ovardystrophie wurden zusätzlich mit einer Eierstocksmassage behandelt. Die Versuchstiere wurden 56 und 72 Stunden nach Entfernung der Spirale blind besamt. Die Brunstin-duktionsrate betrug insgesamt 90,1%, bei denen rnit Ovardystrophie 77,7%. Die An-wendung der PRID-Spirale führte zur Verbesserung der Fertilitätslage. Die Gesamt-trächtigkeitsrate lag mit 87,6% bei den Versuchskühenüber derjenigen der Kontrollkü-he mit 79,3%. Die Behandlung der Tiere rnit Ovardystrophie erbrachte eine Gesamt-trächtigkeitsrate von 88,8% bei den Versuchs- und 72,7% bei den Kontrolltieren. Die Zwischentragezeit betrug bei den Versuchstieren 101,2 Tage und bei den Kontrolltieren 113,3 Tage (p ≤ 0,05). Bei den azyklischen Tieren lag die Zwischentragezeit bei 104,6 Tage bzw. 134,7 Tage (p ≤ 0,05). In einem anderen Versuch wurde 20 Hybriden zwischen Wisent und Hausrind rwecks Zyklussynchronisation die PRID-Spirale verabreicht. Die Brunstinduktionsrate betrug 66,6%, das Erstbesamungsergebnis nach Doppelbesamung 30%, die Gesamtträchtig-keitsrate nach dem Decken der umrindernden Kreuzungstiere rnit einem Bullen 90%. Die Deckperiode konnte verkürzt werden. Contents: Investigations on the synchronization of estrus cycle in cattle from farms with fertility problems and in cross-breeds between bison and cattle with a PRID-de-vice In 81 experimental and 92 control animals of the “Polish Black and White” breed derived from 3 farms with fertility problems an investigation was carried out for sterility prophylaxis using the PRID-device. Based on clinical findings and milk progesterone values cattle with ovarian dystrophy (18 experimental and 22 control animals) from a special group were used. The begin of treatment was between 60 and 80 days post partum. All experimental animals had received a PRID-device for a duration of 12 days. Control animals were injected with physiological saline solution and animals with ovarian dystrophy were treated additionally with ovarian massage. Experimental animals were inseminated 56 and 72 hrs after removal of the device. All animals taken, estrus was induced in 90.1% of the animals, while only 77.7% of animals with ovarian dystrophy came to estrus. Application of the PRID-device led to an improvement of fertility. Pregnancy rates in experimental animals were 87.6%, while only 79.3% of the control animals were pregnant. Treatment of animals with ovarian dystrophy resulted in 88.8% pregnancies for treated and 72.7% for control animals. Experimental animals were non-pregnant for a duration of 101.2 days, while control animals remained non-pregnant for 113.3 days (p < 0.05). Acyclic cattle had a non-pregnant period of 104.6 days and 134.7 days (p < 0.05) respectively. In a second experiment 20 cross-breeds between bison and domestic cattle were treated with a PRID-device for synchronization of the estrus cycle. The rate of estrus induction was 66.6%, the fertility after double insemination 30%, the total pregnancy rate improved after mating with a bull of animals that had not conceived to 90%.     

Influence of various feeding conditions, the migrating myoelectric complex and cholinergic drugs on antral slow waves in sheep.

The presented study was designed to elucidate whether the cholinergic mechanisms control ovine antral slow waves in various physiological conditions, including feeding and various phases of migrating myoelectric complex (MMC). The investigations were carried out on six adult sheep of Polish Merino breed with seven bipolar electrodes surgically implanted onto the antral and small intestinal wall. In the course of chronic experiments, the myoelectric activity was recorded from these regions using the multichannel electroencephalograph. Experiments were performed on 48 h fasted and non-fasted animals. During some of these experiments, sheep were fed with standard fodder. During control experiments 0.15 M NaCl was slowly administered i.v. through the indwelling catheter and during other experiment, hexamethonium bromide (2.0 and 5.0 mg/kg). atropine sulfate (0.02; 0.1; 0.5 and 1.5 mg/kg) and pirenzepine dihydrochloride (0.02; 0.5 and 2.0 mg/kg) were administered i.v. during phase 1-2a or 2b MMC. The drugs were also given in combinations. The recordings were analysed and the antral slow wave amplitudes and frequencies were calculated. Unlike the slow wave amplitude, either feeding or the anticholinergic drugs significantly increased slow wave frequency, especially when the given procedure was started during phase 2b MMC. The most pronounced effects were observed after hexamethonium given alone or in combinations. Thus, the cholinergic system modulates antral slow wave frequency in sheep.     

Differential effects of heparin on NO and tumor necrosis factor-alpha production in bovine blood mononuclear cells stimulated with Salmonella typhimurium lipopolisaccharide.

We investigated the influence of heparin, one of the extracellular matrix (ECM) components, on nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) production by bovine peripheral blood mononuclear cells (PBMC) and monocytes left to adhere for 2 (freshly adherent monocytes) and 48 h (resting monocytes), activated with Salmonella typhimurium lipopolysaccharide (LPS). After 24-h stimulation with LPS, heparin (100 microg/ml) increased (by about 40%) NO production by peripheral blood mononuclear cells and by freshly adherent monocytes. However, it did not change NO synthesis by the resting monocytes. Unlike its influence on NO level, heparin diminished TNF-alpha production by PBMC and monocytes stimulated with LPS. Microscopical examination of PBMC stained with biotin-labeled heparin, showed that both lymphocytes and monocytes were able to bind this glycosaminoglycan. We suggest that heparin, as a component of ECM, modulates the early response of monocytes to exogenous stimuli.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001     

Effect of 4-chloro-2-methylphenoxyacetic acid and 2,4-dimethylphenol on human erythrocytes

The effects of exposure of human erythrocytes to different concentrations of 4-chloro-2-methylphenoxyacetic acid (MCPA) and its metabolite—2,4-dimethylphenol (2,4-DMP) were studied. The investigations concerned mainly the content of glutathione (GSH and GSSG), glutathione peroxidase (GSH-Px), glutathione transferase (GST), and the level of adenine energy charge (AEC). Reactive oxygen species (ROS) such as hydroxyl radical, superoxide anion, hydrogen peroxide, and nitric oxide are produced during normal processes in the cell. Under normal conditions, antioxidant systems of the cell minimize damage caused by ROS. When ROS generation increases to an extent that it overcomes the cellular antioxidant systems, the result is oxidative stress. We observed that MCPA and 2,4-DMP decreased the level of GSH in erythrocytes in comparison with control. MCPA did not affect glutathione peroxidase and glutathione transferase activity, while 2,4-DMP increased their activity. 2,4-DMP decreased the level of ATP and increased the content of ADP and AMP, leading to the fall of the level of AEC. MCPA and 2,4-DMP transform hemoglobin into methemoglobin, thus preventing oxygen transport. Comparison of the toxicity of MCPA and 2,4-DMP revealed that the most prominent changes occurred in human erythrocytes incubated with 2,4-DMP.