Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA)

Squash mosaic virus (SqMV, comovirus) is seed-transmitted in severalCucurbitaceae. Therefore, the use of virus-free seed is important to prevent establishment of this virus in the Netherlands and to avoid spread to other countries.This study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of SqMV in melon seeds. An antiserum was produced to a serotype 1 isolate from melon. Two ELISA variants were investigated viz. an ELISA variant with simultaneous incubation of sample and enzyme conjugate (ELISA 1) and an ELISA variant with successive incubation of sample and enzyme conjugate (ELISA 2). The sensitivity of ELISA was tested by mixing fluor of ground infected and non-infected seeds in different proportions. SqMV was detected by both ELISA variants at dilutions of 1 160 (1 part of infected flour mixed with 159 parts of non-infected flour) or higher after a substrate incubation period of 4 h. However, ELISA 1 gave relatively higher absorbance values than ELISA 2 for nearly all dilutions. Since ELISA 1 is also faster than ELISA 2, ELISA 1 is advised for routine testing. In these test, using subsamples of 100 melon seeds SqMV is detected reliably. ELISA 1 is now used in the Netherlands for routine-indexing of melon seed lots for SqMV.Samenvatting Het pompoenemozaïekvirus gaat over met het zaad van verscheideneCucurbitaceae. Het gebruik van virusvrij zaad is belangrijk om te voorkomen dat het virus zijn intrede doet in Nederland en zich naar andere landen verspreidt.Een antiserum werd geproduceerd tegen een serotype 1 isolaat van meloen. Met behulp van dit antiserum werd een ELISA ontwikkeld om pompoenemozaïekvirus in zaden van meloen aan te tonen. Twee varianten van ELISA werden vergeleken, namelijk een variant waarbij monster en enzymconjugaat gelijktijdig geïncubeerd werden (ELISA 1) en een variant waarbij monster en enzymconjugaat na elkaar geïncubeerd werden (ELISA 2). De gevoeligheid van de ELISA varianten werd uitgetoetst door meel van zieke zaden in verschillende verhoudingen te mengen met meel van gezonde zaden. Het pompoenemozaïekvirus werd met beide ELISA varianten aangetoond in verdunningen van 1 160 (1 deel meel van zieke zaden gemengd met 159 delen meel van gezonde zaden) of hoger na 4 uur incubatie met substraat. ELISA 1 gaf doorgaans hogere extinctiewaarden dan ELISA 2 voor bijna alle verdunningen. Omdat ELISA 1 ook nog sneller is dan ELISA 2, wordt ELISA 1 aanbevolen voor routinematig gebruik. Wanneer voor routinematig gebruik 100 meloenezaden per submonster getoetst worden, kan het pompoenemozaïek virus betrouwbaar worden aangetoond. In Nederland worden momenteel per zaadpartij 20 submonsters van 100 zaden getoetst.     

Taste aversion learning in horses

The ability of ponies to learn to avoid a relatively novel food associated with illness was tested in three situations: when illness occurred immediately after consuming a feed; when illness occurred 30 min after consuming a feed; and when illness was contingent upon eating one of three feeds offered simultaneously. Apomorphine was used to produce illness. The feeds associated with illness were corn, alfalfa pellets, sweet feed and a complete pelleted feed. The ponies learned to avoid all the fees except the complete feed when apomorphine injection immediately followed consumption of the feed. However, the ponies did not learn to avoid a feed if apomorphine was delayed 30 min after feed consumption. They could learn to avoid alfalfa pellets, but not corn, when these feeds were presented with the familiar "safe foods," oats and soybean meal. Ponies apparently are able to learn a taste aversion, but there were constraints on this learning ability. Under the conditions of this study, they did not learn to avoid a food that made them sick long after consumption of the food, and they had more difficulty learning to avoid highly palatable feeds.     

Determination of uric acid in canine serum and urine by high performance liquid chromatography

A reproducible high performance liquid chromatography (HPLC) method was developed for analysis of uric acid in canine serum and urine. The method consists of precipitating serum proteins with phosphotungstic acid prior to HPLC analysis. Urine is analyzed after dilution with buffer. Chromatography is performed on a reversed-phase C-18 column with UV detection at 292 nm. Sensitivity of the method will allow reproducible measurement of uric acid at concentrations of 0.05 mg/dl in serum and 0.1 mg/dl in urine. The HPLC method has been used to quantify hundreds of canine serum and urine samples. The method is superior to UV absorption or colorimetric methods because its lower limit of detection allows measurement of uric acid at concentrations found in canine serum and urine.     

Serum 5'nucleotidase activity in rats: a method for automated analysis and criteria for interpretation

A manual kit for determining serum 5'nucleotidase (5'NT, EC 3.1.3.5) activity was adapted for use with rat samples on a large discrete clinical chemistry analyzer. The precision of the method was good (within-run C.V. = 2.14%; between-run C.V. = 5.5%). A comparison of the new automated method with a manual and semi-automated method gave regression statistics of y = 1.18X -3.66 (Sy. x = 4.54), and y = 0.733X + 1.97 (Sy. x = 1.69), respectively. Temperature conversion factors provided by the kit manufacturer for human samples were determined to be inaccurate for converting results from rat samples. Analysis of components contributing to normal variation in rat serum 5'NT activity showed age and sex to be major factors. Increased serum 5'NT activity was observed in female rats when compared to male rats beginning at about 5 to 6 weeks of age. An analysis of variance of serum 5'NT, alkaline phosphatase, and GGT activities observed over a 9-week period in normal rats suggests several advantages for 5'NT as a predictor of biliary lesions in rats.     

Airway hypersensitivity induced by Ascaris suum extract in New Zealand Romney sheep

Sheep from local farms with and without previous exposure to pigs were tested for their skin and airway responses to a commercial Ascaris suum antigen. There was an immediate reaction to intradermal injection of the antigen in 90% of 101 sheep. A bronchial provocation test by aerosol of the same antigen was undertaken on 43 of the sheep with a positive skin reaction. About 70% of sheep showed an immediate airway response to the antigen as an aerosol, reflected as a significant increase in airway resistance and/or decrease of dynamic lung compliance. The mean peak airway resistance and mean lowest dynamic lung compliance were 165% above and 61% below their baselines, respectively. No significant changes were recorded when the same animals were given an aerosol of phosphate buffered saline. Similarly, no correlation was found between the degree of skin reaction and the magnitude of bronchoconstriction (p>0.05). The sheep with previous exposure to pigs showed no significant differences in airway responses to antigen challenge, although they showed significantly greater skin reactions than those without exposure to pigs. These results indicate that the majority of Romney sheep in the Manawatu have a natural skin and airway sensitivity to A. suum antigen and may therefore be used as an animal model to study human airway hypersensitivity. The origin of this sensitivity has yet to be determined.     

Boar culling in swine breeding herds in Minnesota

This investigation was conducted to study boar culling patterns in swine breeding herds. Data were obtained from 84 swine breeding herds and included 440 boars that were culled. Each producer was involved for 12 consecutive months, and recorded every boar that was removed from the herd, the date, and the reason for culling. The annual culling rate for the 84 farms averaged 59.4% ± 6.4 (SEM). The correlation coefficient between boar and sow culling was 0.52 (p <0.0001). Removal was the result of being overweight (47%), reproductive problems (18%), leg problems (12%), death (7%), and other diseases (4%). From the annual culling rate, the average breeding life of boars was estimated at 20 months. From this study, we concluded that the annual culling rate for boars in commercial herds was high and related to several different factors.     

Nephrotoxicity in dogs associated with methoxyflurane anesthesia and flunixin meglumine analgesia

Uremia unexpectedly developed in five dogs 24 hours after undergoing thoracotomy in a student laboratory. In all dogs general anesthesia had been maintained with methoxyflurane, muscle relaxation had been induced with gallamine, and each dog received a single intravenous dose of 1.0 mg/kg flunixin meglumine for analgesia upon termination of anesthesia. In a subsequent group of dogs undergoing an orthopedic procedure, we assessed the effects on renal function of methoxyflurane anesthesia plus oxymorphone, or of methoxyflurane or halothane anesthesia in combination with a single IM 1.0 mg/kg dose of flunixin meglumine. Significant elevations in serum urea and creatinine values, and necrosis of collecting ducts and loops of Henle, were noted only in the dogs receiving methoxyflurane and flunixin meglumine.

We conclude that the use of combination of methoxyflurane and flunixin meglumine is contraindicated in dogs.

     

An assessment of the copper status of dairy herds in the Waikato, Taranaki and Northland in spring and the effects of daily supplementation with copper sulphate

The copper status of dairy herds in the Waikato, Taranaki and Northland regions between 1 August and 30 September was assessed by copper determinations made on composite sera from ten cows in each herd. Comparisons were made between two major groups categorised as as received and supplemented. In the latter, the cows received 2.0-6.0 g/day of copper sulphate orally from late gestation to the end of lactation. Mean copper values were determined from pastures in Taranaki and Northland and from pastures from three soil types in the Waikato. The lowest were in Northland (134.3 micromol/kg) and the highest in Taranaki (173.3 micromol/kg). Calculated dietary available copper levels showed the lowest were from pastures grown on organic soils in the Waikato. These soils also had high molybdenum levels. Serum copper values as received were lowest in Northland (7.30 micromol/l), and were marginal for both Taranaki (9.91 micromol/l) and the Waikato (9.93 micromol/l). The levels in supplemented herds were considerably higher, i.e. from the Waikato 11.7 micromol/l and from Taranaki 11.5 micromol/l. When liver and serum copper levels from paired samples were compared there was a reasonable correlation (r = 0.64) but the standard deviation approximated the mean liver copper level, i.e. 155 +/- 141 micromol/l.