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.     

Teaching animal welfare in the land grant universities

Colleges and universities have an obligation to teach the basis of animal husbandry and welfare and must prepare students so that they can respond effectively to challenges by proponents of the animal welfare and animal rights movements. Veterinary curricula must now contain formal instruction in professional ethics and humane stewardship of animals for accreditation by the American Veterinary Medical Association. It is helpful if students have an understanding of farm animal behavior, stress physiology and methods of assessing welfare prior to learning about the animal welfare/rights movement's philosophies and issues. A review of early judicial practices, "classical" Judeo-Christian philosophy, the philosophy of Rene Descartes, Jeremy Bentham, Albert Schweitzer, and current philosophers and the entertainment media places the movements in perspective. Students should be familiar with such concepts as the mind-body controversy, equality of suffering, self-awareness or intelligence, and speciesism. After acquiring an appreciation of the basics, a knowledge of the issues facing animal agriculture and the arguments for and against each issue are necessary. Graduates of colleges of agriculture need to realize the potential effects the movements can have and take the initiative to improve the image of animal agriculture.     

Detection of a novel chloramphenicol resistance plasmid from "equine" Staphylococcus sciuri

A small chloramphenicol resistance (Cm) plasmid of 4.65 kB could be detected in an "equine" Staphylococcus sciuri-culture. This plasmid, designated as pSC3, was identified by interspecific protoplast transformation. On the basis of restriction endonuclease analyses a detailed restriction map of pSC3 could be constructed. This allowed structural comparisons of pSC3 with Cm-plasmids of other staphylococcal species from infections of humans and animals and identification of pSC3 as a member of the pC 221-family of staphylococcal Cm-plasmids. The pSC3-plasmid encoded an inducible chloramphenicol acetyltransferase as confirmed by enzymatic assays. This enzyme could be demonstrated in cell-free lysates of Cm-induced pSC3-transformants.     

30 years of swine health services in Bavaria

A Pig Health Service in Bavaria was established in 1958, there was one veterinary-officer working. The department Pig-Health Service of the "Tiergesundheitsdienst Bayern" has twenty-one veterinary-officers now. They mostly are working in the pig health scheme, doing stress susceptibility test, giving advice and expert evidence.     

Plasmid-encoded antibiotic resistance in Staphylococcus hyicus

A total of 33 Staphylococcus hyicus-cultures from piglets with exudative epidermatitis were analyzed for the presence of antibiotic resistance plasmids. Four small plasmids encoding resistances to chloramphenicol, macrolide-lincosamide-antibiotics, streptomycin or tetracyclines could be identified in plasmid-curing and plasmid-transformation experiments. For further characterization these plasmids were digested with restriction endonucleases. This led to the construction of a specific restriction map for each of the 4 plasmids. On the basis of their restriction maps, these 4 antibiotic resistance plasmids from "porcine" S. hyicus-cultures were compared with the respective resistance plasmids of other staphylococcal species from infections of humans and animals.     

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.     

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.

     

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.