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.     

Exclusion of infectious diseases from sheep and goat farms

It is important for practitioners and owners to be aware of the risk associated with an open herd. This article has discussed some of the infectious diseases that might be inadvertently introduced. Many others--including most exotic diseases and some that are regulated by governmental agencies, such as tuberculosis and brucellosis--have been omitted. It should be obvious that a simple health certificate is an inadequate guarantee of freedom from infections. Instead, producers should be prepared to invest both time and money in evaluating the health status of the herd of origin. Some of the diseases discussed, such as foot rot, cause tremendous financial losses. Others, such as pinkeye, can be easily "lived with." The point to be made is that producers should consider all these and other diseases and then make informed decisions based on the probable cost of these diseases if they became established in the herd.     

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.     

Salmonella isolated from feeds and feed ingredients during the period 1982-1988: animal and public health implications

The prevalence of Salmonella in southern Africa in farm feeds and by-products of animal origin during 1982-1988 was determined. Salmonella occurred in 5.18% of the farm feed samples and in 9.54% of the by-product samples. Different serovars were isolated, some only once. The findings underestimate the true prevalence of Salmonella in farm feeds and by-products, and is representative of only the most severely contaminated products. The epidemiology of salmonellosis is discussed with special reference to the importance of multiple resistance to antibiotics, the increase in the number of cases of salmonellosis worldwide and "Salmonella free" feeds and foods. More detailed research on the role of farm feeds in the epidemiology of salmonellosis is required. Efforts should be made to increase awareness of the problem, to improve quality management at farm feed production plants and to develop efficient systems to monitor the hygienic safety of feeds and foods.     

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.