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.     

MAGNETIC RESONANCE IMAGING OF THE EQUINE STIFLE

The purpose of this study was to define normal gross anatomic structures in the equine stifle with magnetic resonance images. Magnetic resonance (MR) images were made in sagittal, 15° supinated, transverse, and dorsal planes of two equine stifles. The MR images were scrutinized by comparing MR images to dissection specimens and frozen cross sections of stifle joints. Sagittal and 15° supinated images were the most valuable in assessing articular cartilage, subchondral bone, and soft tissue structures within the joint. Cranial and caudal cruciate ligaments, medial and lateral menisci, meniscotibial and meniscofemoral ligaments, long digital extensor tendon, and patellar ligaments were easily evaluated. MR images provided substantially more gross anatomical information than the currently available imaging modalities.     

Evaluation of Sperm-Activating Solutions in Atlantic Salmon Salmo salar Fertilization Tests

The use of saline solutions was tested to determine their efficacy as replacements for ovarian fluid as sperm activators and to eliminate variability encountered with the use of Ovarian fluid. We tested fertilization rate of semen from eight males on Atlantic salmon Salmo salar eggs after five sperm-activating solutions and a non-activating saline were substituted for ovarian fluid. We used solutions shown acceptable for use with other salmonid species. The six solutions tested were a non sperm-activating phosphate-buffered saline (PBS, 7.2 g/L NaCl, 1.48 g/L Na₂HPO₄, 0.43 g/L K H₂PO₄), a Tris buffer (6.99 g/L NaCl, 3.63 g/L Tris and 2.42 g/L glycine), a Borax buffer (12.2 g boric acid/L in solution 1, 76 g disodium tetraborate/L combined 100:118, then 1 L combined with 3.7 L water and 18 g NaCl), and three solutions of 9.25 g/L (125 mM) NaCl buffered to pH 6.0, 7.5, and 8.9. The latter five solutions activated sperm immediately on contact, while PBS required additional water to activate sperm. The PBS solution was the least effective (mean percent eyed eggs 37.6%) for egg fertilization. The mean percent eyed eggs for the other five saline solutions (range 78% to 86%) were not significantly different. Sperm from one male provided significantly lower egg fertilization (39.6%) when compared with the other seven males (67.2–87.4% egg fertilization). Percent egg fertilization was not related to number of live sperm cells per egg. Our results show that osmotically-balanced sperm-activation solutions, even those with a pH range from 6.0 to 8.9 provide adequate media for fertilization of Atlantic salmon eggs. Fertilization in a deactivation saline and water reactivation of sperm resulted in low egg fertilization.     

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.     

Caprine beta-Mannosidosis in Kids from an Ontario Herd

Caprine β-mannosidosis, a fatal inherited deficiency of the lysosomal enzyme β-mannosidase, was diagnosed in neonatal female Nubian crossbred twin kids from a small herd near Guelph, Ontario. The kids had been tetraplegic since birth, with whole body tremors, abnormal nystagmus and an intention tremor of the head.     

Resistance to yellow rust in Triticum dicoccoides. II. Crosses with resistant Triticum dicoccoides sel. G-25

A comparison was made between the genes in 29 new selections of wild emmer wheat resistant to yellow rust over wide geographic areas and the previously extensively studied selectionTriticum dicoccoides G-25. In 23 selections the resistance may be conferred by 1 dominant gene; these include 11 selections in which the gene is different from the dominant gene in sel. G-25 and two others in which the genes were closely linked or allelic to the gene in G-25, differing from sel. G-25 by race-specificity. Two dominant genes different from the gene in sel. G-25, seem to be present in one selection. In five selections the resistance may be conferred by one or two recessive genes, including three instances in which the recessive gene was associated with a dominat gene. Our findings show that at least 19 out of the 29 selections studied possess genes which are different from the gene inT. dicoccoides sel. G-25.Samenvatting In dit onderzoek werden 29 nieuwe resistente wilde-emmer selecties (Triticum dicoccoides) gekruist met de reeds uitvoerig bestudeerde resistente selectie G-25, om na te gaan of de resistentie van de nieuwe selecties wordt veroorzaakt door genen op dezelfde locus als het dominante gen in sel. G-25 of dat er andere loci bij zijn betrokken. De ouders, de F₁-en F₂-populaties van een bepaalade selectie werden in het kiemplantstadium getoetst met één Israëlisch gele-roest isolaat van fysio 2E0 of van fysio 2E18. In de uitsplitsende F₂-populaties werden de niet-sporulerende planten als resistent beschouwd en de sporulerende als vatbaar.In de F₂-populaties van 12 herkomsten werden geen vatbare planten gevonden, hetgeen er op duidt dat de resistentie wordt veroorzaakt door een gen op dezelfde locus als het gen in G-25 of door een gen dat neuw gekoppeld is aan het gen in G-25. Voor twee van deze herkomsten kan op basis van een fysio-specifieke interactie worden vastgesteld dat de resistentie berust op allelen die verschillen van het allel in sel. G-25. In 11 herkomsten werd een uitsplitsing voor twee dominante gene gevonden (RS=151), waarbij het tweede dominante gen uit de getoetste nieuwe selectie afkomstig is. De aanwezigheid van twee dominante genen verschillend van het gen in sel. G-25 werd gevonden in één herkomst (631). In de overige vijf selecties bleek de resistentie te worden veroorzaakt door één of twee recessieve genen waarnaast in drie gevallen ook nog een dominant gen werd gevonden.De resultaten tonen aan dat tenminste 19 van de 29 bestudeerde selecties resistentiegenen bezitten die verschillen van het gen inT. dicoccoides sel. G-25. Slechts in twee van deze selecties kan het gen allel zijn met het gen in sel. G-25.