Test for linkage between the eumelanin dilution blue (B1), the extended black (E) allele at the E-locus and the linked pea comb (P) and eumelanin extension (M1) genes in the domestic fowl

1. A mating was made between a Blue Andalusian bantam male and a pea comb striped necked bantam female to produce a quadruple heterozygote of the linked pea comb (P) and eumelanin extension (Ml) genes and of the eunmelanin dilution blue (Bl) gene and the extended black (E) allele at the E-locus. 2. Two pea comb blue females so produced were mated to a single comb striped necked bantam male, thus providing a backcross to the quadruple recessive. 3. Bl segregated independently with each of the other three genes whilst 25 crossovers occurred amongst the 55 progeny assessed for linkage between P and Ml, confirming loose linkage between P and Ml which had previously been shown to be approximately 46 units. Linkage of 0.3% had been previously reported (Crawford, 1986) between P and a "melanotic" gene isolated from a segregating gene pool. Evidence is presented to demonstrate that two different "melanotic" genes were being considered, thus explaining the apparently contradictory reports. Charcoal (cha) is suggested for the recessive gene linked with P by 0.3%. 4. Thirty-four crossovers occurred among the 97 progeny between P and E suggesting linkage of approximately 35 units. A further mating was made from among segregants in the backcross which confirmed P-E linkage with 17 crossovers in 55 gametes. Combining the tests produced a linkage value of approximately 34 units.     

Influence of the sex-linked dwarfing gene (dw) on the contribution of dietary lipid to yolk lipid synthesis

1. The efficiency and time course of dietary fatty acid incorporation into lipids of egg yolk and abdominal adipose tissue was compared in "White Leghorn", normal (Dw) and dwarf (dw) laying hens at 56 weeks of age, using 14C labelled linoleic acid. 2. The sex-linked dwarfing gene, dw, was shown to reduce not only body weight and abdominal fat pad deposition, but also yolk production and the average clutch size. 3. Higher peak incorporation and total recovery of the linoleic acid radioactivity into yolk lipids, but lower label recovery into adipose tissue triglycerides were found in dwarf hens. 4. The higher esterification of the dietary linoleic acid in its native form into dwarf yolk triglycerides indicates that dwarf hens use more dietary lipids to synthesise yolk lipids but these results also suggest that the dw allele might reduce the lipogenic capacities of the liver and adipose tissue in laying hens.     

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.     

Calving patterns in seasonal dairy herds

Two field studies examined the calving patterns of cows in seasonal dairy herds in the Waikato (Field Study 1) and South Taranaki regions (Field Study 2). The first study examined patterns for cows commencing their second or subsequent lactation in herds which had used an inseminating service during the previous season. The second study included first lactation heifers only in 15 herds where animals had been naturally mated, and in 15 herds in which they had been synchronised and then artificially inseminated at the synchronised oestrus. The parameters describing calving patterns were based on the date for each herd's planned start of calving (PSC), which was 282 days from the date on which breeding commenced in the preceding season. The average interval from PSC to mean calving date for the 35 herds in Field Study 1 was 22 days, with individual herds ranging from 15 to 30 days. In herds with heifers which had been naturally mated (Field Study 2), it was 17.6 days compared to 11.0 days for previously synchronised animals. Calculating the intervals from PSC to median calving date and separately for the last two quartiles more effectively described a herd's calving pattern. The duration for the last quartile of the calving pattern was influenced by the extent and timing of induced calving. In Field Study 1, 88.6% of the 35 herd owners induced premature parturition in at least one cow. In these herds, 11.3% of cows were treated and calved prematurely. Only 61.7% of heifers which had previously been naturally mated calved by 3 weeks after PSC. Their calving dates were not evenly distributed over this 3-week period, with 9.8% in the first week and 25.6% in the third week. The calving pattern for heifers which had been previously synchronised showed several distinct peaks. Calvings to the synchronised mating were completed 15 days after PSC, by which time 64.7% of animals had calved. By 3 weeks after PSC, 72.9% of these heifers had calved. The results showed that there was considerable variation in calving patterns in seasonal dairy herds. This variation would have been due to differences in conception pattern, and the way induced calving had been applied. The calving pattern in heifers which had been naturally mated was less concentrated than had been expected. Synchronisation can significantly concentrate the calving pattern of these first lactation animals. The parameters used to describe calving patterns may be less applicable in herds in which a high proportion of animals is induced to calve prematurely, or where a whole herd is synchronised. Nonetheless, they do serve as an illustrative example of the variation in calving patterns among herds.     

Epizoologic studies of the detection of antibodies against Aujeszky's disease virus in sera and blood eluates of swine from Thailand using ELISA ("Enzygnost," Behring), serum neutralization test and "Aujeszky Latex Kit" (Iffa Merieux)

The results of three tests for Aujeszky's disease were analysed and compared. The presence of Aujeszky's antibodies was determined by "Enzyme-linked-Immunosorbent-Assays" (ELISA, "Enzygnost"), Behring company, Marburg; "Serum-Neutralization-Tests" (SNT); and "Latex Agglutination-Tests" (LT, "Aujeszky-Latex-Kit"), Iffa Merieux company, Laupheim. Whole blood and sera samples were taken from 805 swine from 26 of Thailand's provinces. These samples were analysed to determine if eluates of whole blood on filter paper were as effective as corresponding sera samples in determining the presence of Aujeszky's disease antibodies. From a total of 805 samples, 26% of the serum and 18% of the blood eluate samples showed a positive result when tested by the ELISA method. Clearly, testing whole blood eluates provides results which are inferior to results from sera samples. Therefore the ELISA whole blood eluates test can only be recommended with reservations. Further testing was done on 645 serum samples using SNT. Samples tested were those which gave negative, suspicious, or weakly positive results when tested by ELISA. Using SNT, 23% of these showed a positive result. Many serum and blood eluate samples were also tested by LT. Most of these test samples were chosen because they were deemed suspicious. Suspicious samples were defined as those which had deviant test results. According to these results the sensitivity of LT was between the sensitivity of SNT and ELISA. Owner survey responses tended to state that few animals had been vaccinated. This coupled with the frequency of antibody occurrence proves the high rate of infection among Thailand's swine population.