Use of monoclonal antibodies against human HLA II antigens for the detection of bovine B lymphocytes and macrophages]

The crossreactivity of mouse monoclonal antibodies (MoAbs) (Tab. I) prepared against human HLA-DR and HLA-DP antigens was studied in various bovine cells: lymphocytes from lymph nodes and peripheral blood, adherent (B) and nonadherent (T) lymphocytes, monocytes, granulocytes and platelets. In the immunofluorescence test, MoAbs Bra13, Bra14, Bra20, Bra22, Bra30, Bra70, HL-38 reacted with bovine B lymphocytes and monocytes, but not with other tested cells (Tab. III, IV). These antibodies, except Bra22, were positive with B lymphocytes in the complement dependent cytotoxic test (Tab. II). The similarity of the bovine antigens and HLA-DR antigens determined by used MoAbs was also proved by immunoblotting. Monoclonal antibodies Bra38 and BraFB6 did not react with the bovine cells and separated antigens. The epitope (HLA-DR) recognized by the antibody Bra38 is probably absent in cattle. The presence of HLA-DP analogue determined by the antibody BraFB6 has not been confirmed. The crossreactive MoAbs could be used for the detection of B lymphocytes and macrophages in veterinary immunology.     

An evaluation of the Olsen test as a measure of plant-available phosphorus in grassland soils derived from basalt parent material

Abstract. Anecdotal and circumstantial evidence have suggested that the Olsen test underestimates plant-available phosphorus (P) in basaltic soils in Northern Ireland. Therefore, the ability of this test to predict plant-available P in basaltic (and non-basaltic) soils was investigated by regressing Olsen-P data against herbage P indices calculated from plant tissue test data using the diagnosis and recommendation integrated system. The average Olsen-P concentration for a range of fields situated on basaltic soils was considerably lower than the average Olsen-P concentration for a range of fields situated on non-basaltic soils, and yet mean sward P status, as given by the herbage P indices, was similar for both groups of fields. Herbage P indices were also much better correlated with Olsen-P measurements in non-basaltic soils than in basaltic soils. Furthermore, at low Olsen-P values (≶9mgPL⁻¹) some swards on basaltic soils were genuinely deficient in P, while others were sufficient or even in surplus for this nutrient. The results confirm that Olsen-P is inadequate as a predictor of plant-available P in basaltic soils. It is concluded that an alternative soil test is needed to provide a reliable assessment of plant-available P in basaltic soils, to prevent overuse of fertilizer and manure P and to minimize the amounts of P entering local watercourses.     

Identification of the porcine cytomegalovirus major capsid protein gene.

A major capsid protein (MCP) gene homologue of porcine cytomegalovirus (PCMV) was identified. Sequence analysis indicated that the PCMV MCP gene is 4,026 nucleotides in length encoding a protein of 1,341 amino acid residues. The predicted molecular weight of the PCMV MCP is 151,456 Da, equivalent to those of other herpesvirus MCP counterparts. Phylogenetic analysis using herpesviral MCP gene sequences confirmed that PCMV is a betaherpesvirus with higher homology with human herpesvirus-6 and -7 than human and mouse cytomegaloviruses. The serum of pig experimentally infected with PCMV did not react with bacterially expressed MCP, suggesting that the PCMV MCP may not be related to the humoral immune response in the course of PCMV infection. Also, we established polymerase chain reaction (PCR) protocols using primers corresponding to MCP gene sequences for detection of PCMV infection. The PCR protocol would be effective for the diagnosis of slow-growing PCMV infection, for which traditional methods involving virus-isolation are not useful.     

Doxorubicin affects the cardiac muscarinic system in the rat.

During the study on the mechanism of doxorubicin-induced cardiotoxicity, we observed that a long incubation (4 hr) with doxorubicin reduced the maximal negative inotropic effects of a muscarinic receptor agonist, carbachol. The mechanism responsible for this doxorubicin-induced reduction of the efficacy of carbachol was examined in isolated guinea pig hearts. In isolated left atrial muscle preparations, 1 hr incubation with 100 microM doxorubicin caused a parallel right-ward shift of the concentration-response curves for carbachol, but a longer (4 hr) incubation with this agent (30, 100 or 200 microM), caused a significant reduction of the magnitude of the negative inotropic effect of carbachol in addition to the concentration-dependent parallel right-ward shift. The 4-hr incubation with these concentrations of doxorubicin also reduced the maximal negative inotropic effect of an adenosine A1 receptor agonist, R-phenylisopropyl adenosine (R-PIA), without affecting the potency of this agonist. Doxorubicin (1 to 100 microM) reduced [3H]quinuclidinyl benzilate (QNB) binding in a concentration dependent manner, but failed to alter [3HIR-PIA binding. The decrease in the magnitude of the maximal negative inotropic effect by doxorubicin was caused by changes in the muscarinic system at steps common to the transduction of muscarinic and adenosine A1 receptor mechanisms.     

The extent of genetic diversity among Vanilla species: Comparative results for RAPD and ISSR

Vanilla is a large genus of about 110 species in the orchid family (Orchidaceae), including the species Vanilla planifolia from which commercial vanilla flavoring is derived. Since most species of vanilla are considered rare and endangered there is an urgent need to conserve them through genetic analysis and propagation/conservation studies on this crop.The present study investigated the genetic diversity among nine leafy- and leaf-less Vanilla species employing 30 decamer RAPD primers and 10 ISSR primers. The species under study were diverse and displayed a range of variability (0–66% and 0–81% for RAPD and ISSR, respectively). A total of 154 RAPD polymorphic markers (83.24%, h = 0.378) and 93 ISSR polymorphic markers (86.11%, h = 0.363) were used to generate a genetic similarity matrix followed by the cluster analysis. Specific groupings were revealed by each cluster analysis with slight variation between two different markers. Among the nine species studied, V. planifolia, Vanilla aphylla and Vanilla tahitensis revealed very low level of variation within their collections, thus indicating a narrow genetic base. The large genetic distance of Vanilla andamanica from other species suggests its different origin. A close genetic affinity was observed between the pairs V. planifolia, V. tahitensis and Vanilla albida, V. aphylla. These are the first comparative results for RAPD and ISSR reporting inter-relationship among nine cultivated, wild and hybrid Vanilla species.     

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%.     

Comparison of three in vitro techniques to estimate benzimidazole resistance in Haemonchus contortus of sheep

Three in vitro assays to detect benzimidazole resistance, namely, the egg-hatch assay, tubulin-binding assay, and a larval-development assay, were evaluated by estimating the level of benzimidazole resistance in three field isolates of Haemonchus contortus compared with a susceptible reference strain. Comparisons were also made with estimates of benzimidazole resistance of the three field strains obtained from an in vivo controlled anthelmintic efficacy test. All three in vitro tests showed similar, consistent results which also suggested greater sensitivity than the in vivo assay. These results indicate that selection of an in vitro technique to determine benzimidazole resistance should therefore be based on considerations other than precision, such as technical expertise, availability of equipment, cost and speed in which diagnosis is required.     

Anesthesia for patients with severe systemic debilitation.

Any patient presenting for anesthesia and surgery who has a severe debilitating problem is at great risk for a prolonged recovery, renal failure, or hypotension. A stable hemodynamic state should be maintained. Fluid homeostasis should be ensured. Anesthetic induction techniques using narcotics and benzodiazepines are my preferred approach, with maintenance of anesthesia incorporating isoflurane, narcotics, or both or the use of regional anesthesia.     

Diagnosis of sarcocystosis in sheep using the indirect fluorescence test and ELISA]

The indirect fluorescent antibody test (IFAT) was compared with the enzyme-linked immunosorbent assay (ELISA) for the detection of specific antibodies to Sarcocystis sp. A set of 275 ovine blood samples was examined by both reactions. Cystozoites of Sarcocystis gigantea were used as the corpuscular antigen for the IFAT. For the diagnostics of sarcocystosis by the ELISA technique used the sandwich test of the antibody titration with a soluble antigen which was also prepared from S. gigantea macrocysts. Our studies confirmed that this antigen did not cross-react with Toxoplasma gondii. Titre 40 was determined as the limit one for the IFAT and titre 80 for the ELISA; which was confirmed by the direct detection of cysts in the muscles (Svobodová, 1989). The results of both methods are shown in Table I. 76.7% of the blood samples reacted positively in the IFAT and 83.6% in the ELISA. These methods were found to be suitable and can be utilized for the intravital routine diagnostics.