The heterogenicity of Cowdria ruminantium stocks: cross-immunity and serology in sheep and pathogenicity to mice

Ten stocks of Cowdria ruminantium (Ball 3, Breed, Comoro, Germishuys, Kümm, Kwanyanga, Mali, Mara, Nonile and Welgevonden) were compared from a cross-immunity, serological and mouse pathogenicity point of view. They were found to differ in varying degrees. Except for the Ball 3, Comoro and Germishuys stocks that were similar but not identical, there was no pattern in the antigenic diversity of the 10 stocks. The Welgevonden stock emerged as the stock that elicits an immunity against most of the South African stocks. The inability of the reference Ball 3 stock to protect sheep against no fewer than 6 other stocks questions the advisability of retaining this stock as the vaccine stock. The antigenic diversity of the 10 stocks could not be correlated with the antibody levels detected with the indirect fluorescent antibody test, since the sera against all 10 stocks reacted positively to the Kümm stock antigen and the variation in titres was not stock-related.     

Toxicity of Haemophilus pleuropneumoniae to porcine lung macrophages

Viable Haemophilus pleuropneumoniae bacteria were toxic for porcine alveolar macrophages in vitro. This cytotoxic effect proved to be dose-related. A cell-free extract of H. pleuropneumoniae, heat-killed bacteria, and a Pasteurella multocida field strain were nontoxic. When macrophages were cultured with H. pleuropneumoniae bacteria in a ratio of 100 macrophages to six bacteria, ultrastructural signs of cellular degeneration were observed within 1 h. This degeneration was observed in macrophages with or without phagosomes containing H. pleuropneumoniae. A cytotoxic substance was filtered from a H. pleuropneumoniae culture in Eagle's minimal essential medium supplemented with Earle's salts (EMEM) and 10% foetal calf serum that was incubated for 10 h at 37 degrees C. This substance was destroyed by heating at 65 degrees C for 30 min. Macrophages were less susceptible to the toxic effect of H. pleuropneumoniae when serum of convalescent pigs was added.     

A COMPARISON OF DIATRIZOATE* AND IOXAGLATE† FOR POSITIVE CONTRAST SHOULDER ARTHROGRAPHY IN DOGS

In six experimental dogs, arthrographic quality and synovial inflammatory response with shoulder arthrography comparing meglumine-sodium diatrizoate (Urovison) and the monoacidic dimer, meglumine-sodium ioxaglate (Hexabrix) was evaluated. In our study initial films were of equally high diagnostic quality for both contrast media, but delayed films significantly favored ioxaglate for diagnostic quality. The rise in white blood cells in synovial fluid samples collected 24 hours after the arthrographic procedure was significantly lower after the use of ioxaglate. Histologic examination performed 14 days after the intra-articular injection revealed no drug related lesions.     

Controversial issues in drug treatment during cardiopulmonary resuscitation.

The goal of advanced life support in CPR must be to restore and maintain respiratory and hemodynamic effectiveness, and to correct the underlying dysrhythmia. Optimal basic life-support techniques must be continued to meet these goals. Many drugs have been suggested in the treatment of cardiac arrest, but unfortunately, drug effects are inconsistent and resuscitation rates remain low. Epinephrine, atropine, lidocaine, bretylium, and naloxone remain important drugs for consideration in CPR in most animals with cardiac arrest. The best chance of survival remains in early recognition of animals susceptible to arrest and in treatment of the underlying cause.     

Flow cytometric analysis of T cell response in mice infected with Cowdria ruminantium.

A 3-fold increase in the numbers of Lyt-2+ T cells in the circulating blood of mice infected and re-infected with the Welgevonden stock of Cowdria ruminantium, as determined by flow cytometry, is supportive evidence that immunity in heartwater is cell-mediated. The rise in Lyt-2+ cells only after re-infection of the mice is further evidence that the development of immunity in heartwater is dependent on the unhindered and adequate replication of C. ruminantium.     

Uterine motility of the sow during the oestrous cycle and early pregnancy

With electromyography and strain gauges the uterine motility of the sow during the oestrous cycle and early pregnancy was studied. Special attention was paid to characterization of myometrial activity at the time of intrauterine migration of blastocysts.From recordings of 4 animals (3 became pregnant) 3 types of electrical spiking activity (each could be correlated with an elevation of the strain gauge tension) were discerned. Two of them appeared regularly and were suitable for analysis: High Voltage Slow Acticity (with relative high amplitude and low frequency of spikes) and Low Voltage Fast Activity (with relative low aplitude and high frequency of spikes).The sexual status markedly influenced myometrial activity: during prooestrus and oestrus only one type of electrical activity was present whereas the myoelectrical complex (an episode of this activity and the subsequent interval of inactivity) was shorter than 10 min. During dioestrus the myoelectrical complex was longer than 10 min and High Voltage Slow Activity (solely on the cervix and bifurcation) and Low Voltage Fast Activity occurred simultaneously in episodes which mainly appeared to originate on the bifurcation.The characteristics of uterine activity during pregnancy were similar to those of a cyclic sow until day 12. It was only on day 12 that gestation appeared from an increased frequency of myoelectrical complexes.It is concluded that Low Voltage Fast Activity as it was found on the uterine horn at days 8–9 might be involved in the process of intra-uterine migration of blastocysts. In cyclic and in pregnant animals the patterns of Low Voltage Fast Activity were similar. Therefore, the occurrence of Low Voltage Fast Activity is independent of the actual presence of blastocysts. It seems to be exhibited in dependence on the ovarian hormones.     

Development of a semi-selective medium and an immunofluorescence colony-staining procedure for the detection of Clavibacter michiganensis subsp. sepedonicus in cattle manure slurry

Various compounds and basal media were tested for their suitability to create a semi-selective medium for isolation ofClavibacter michiganensis subsp.sepedonicus (Cms) from cattle manure slurry containing c. 10⁸ colony forming units (cfu) per ml.Plating efficiency of Cms in yeast glucose mineral medium (YGM) was 104% compared with yeast peptone glucose medium. Nalidixic acid, polymyxin B sulphate and the experimental disinfectant S-0208 inhibited colony growth of cattle slurry bacteria as compared with Cms in YGM. The optimal concentration of these inhibitors in combination was determined by modified agar diffusion tests and by pour plating in 24-well tissue culture plates. The semi-selective medium YGMI consisted of YGM supplemented with nalidixic acid (2 mg/l), polymyxin B sulphate (30 mg/l) and S-0208 (125 mg/l). Plating efficiency varied for Cms between 50.9 and 69.6%, for cattle slurry bacteria between 1.8 and 2.5% and for saprophytes from potato heel end extracts between 11.5 and 27.4%.Differentiation of Cms colonies from other colonies was based on their small and bluish colony morphology in pour plates and on immunofluorescence colony-staining (IFC). IFC of a pure culture of micro colonies of Cms in YGM was possible after one day incubation (colonies c. 5 cells). Green background fluorescence in the agar gels was prevented by addition of Tween 20 (0.1%) to the washing buffer and the use of 1% agar gels. IFC of macro colonies of Cms in YGMI, visible with 4x objective magnification, was possible after 4 days. The detection level of the target organism in artificially inoculated cattle slurry in YGMI based on colony morphology varied between 1.4×10³ and 2.3×10⁴ cfu per ml of cattle slurry. Miniaturized plating combined with IFC, using wells in tissue culture plates (=16 mm), proved suitable for detection, but was c. 30 times les sensitive. The recovery of Cms was negatively correlated with the number of saprophytic colonies in the agar plates (R ²=0.74).     

Spring invasion of Heterodera schachtii in sugarbeet; a simulation study

A simulation model was developed for the spring invasion of the beet cyst nematode,Heterodera schachtii Schmidt, into sugarbeet roots, according to the state variable approach. This model describes the processes of egghatch, emergence of second stage larvae from cysts, migration to roots and penetration into roots quantitatively, using published data.In 1983 a field experiment was conducted to test this model.H. schachtii cysts were introduced at depths 6–29 cm in PVC-cylinders, buried in the soil. The rooting depth of sugarbeet seedlings, growing in these cylinders, was limited to 5 cm by 50 m mesh nylon gauze. Every 10 days the second stage larvae, which had penetrated into the roots of these seedlings were counted. After 50 days, about 40% of the eggs had hatched. More than 20% of the emerged larvae penetrated if the cysts had been buried undeeply, and only 4% if the cysts had been buried at 29 cm depth.The model predicted the course of penetration into the root during the first 40 days with reasonable accuracy (r²=0.79), but in the 5th period of 10 days the model made an overestimation of more than 100%. Egghatch after 50 days was correctly simulated. The differences in penetration into the root between the model and the experiment might result from an oversimplified simulation of the penetration success or the neglection of mortality of second stage larvae. Detailed experiments should be done to provide better parameters for these factors.Samenvatting Volgens de toestandsvariabele-benadering werd een simulatiemodel ontwikkeld van de voorjaarspenetratie van het bietecystenaaltje. Het model beschrijft aan de hand van literatuurgegevens het uikomen van de eieren, het verlaten van de cyst door de larven, de migratie naar en de penetratie in de wortel.In 1983 werd een veldproef uitgevoerd om het model te toetsen. Cysten vanH. schachtii werden op 5 dieptes tussen 6 en 29 cm ingegraven in PVC-cylinders, welke waren verzonken in de bodem. De bewortelingsdiepte van de suikerbiete-zaailingen die hierin groeiden werd beperkt tot 5 cm door nylon gaas van 50 m maaswijdte. Elke 10 dagen werden de larven geteld die in de wortels van deze plantjes waren gepenetreerd. Na 50 dagen was 40% van de eieren uitgekomen. Meer dan 20% van de gelokte larven penetreerden als de cysten ondiep waren ingegraven, en slechts 4% als de cysten op 29 cm diepte waren ingegraven.Gedurende de eerste 40 dagen werd het verloop van de penetratie in de wortel met redelijke nauwkeurigheid door het model voorspeld (r²=0.79). In de 5e periode van 10 dagen maakte het model echter een overschatting van meer dan 100%. Het uitkomen van de eieren werd correct gesimuleerd. De verschillen in penetratie tussen het model en de proef zouden het gevolg kunnen zijn van een oververeenvoudigde simulatie van het penetratiesucces of van het verwaarlozen van de mortaliteit van de migrerende larven. Betere gegevens hierover zullen moeten komen uit detailproeven.