An automated micro-method for the enzymatic determination of D-B-hydroxybutyrate in ruminant plasma

1. An enzymatic ‘reaction rate’ micro-method for the rapid routine estimation of D-B-hydroxybutyrate (D-B-HOB) in ruminant plasma, using an I.L. Multistat III centrifugal analyzer, is described.
2. Reaction conditions were optimized to give a linear response for plasma D-B-HOB concentrations between 100 and 2500 μmoles per litre, at 30°C and pH 9.0.
3. For the standardized method the within-run and between-run coefficients of variation for deproteinised ovine plasma were consistently less than 3.5%.
4. There was good agreement between plasma concentrations obtained by the present method and both original U.V. end-point technique (r=0.927b=0.950) and a colorimetric end-point procedure (r=0.937. b=0.879).
5. Utreated ovine and bovine plasma consistently exhibited high ‘blank’ activity and this was directly correlated with plasma lactate dehydrogenase (LDH) activity in both species (r=0.971; p<0.001 and r=0.949; p<0.001 respectively). The distribution of LDH activity in man was similar to sheep but, contrastingly, non-specific interference was extremely low in human plasma and unrelated to LDH. Horse, chicken and rat had negligible ‘blank” activity and comparatively low LDH levels. In both cattle and sheep non-specific interference was abolished by perchloric acid precipitation. In the sheep subtraction of ‘blank’ activity gave D-H-HOB concentrations for untreated plasma comparable to those in deproteinised samples. However, in the bovine, D-B-HOB levels remained significantly (t=6.44; p<0.001) higher even after ‘blank’ correlation. In contrast to man and other non-ruminants, perchloric acid precipitation is essential in ruminants to avoid false overestimation of plasma D-B-HOB levels.
6. Plasma with EDTA as anticoagulant and serum gave concentrations of D-B-HOB approximately 60% lower, than samples containing heparin or oxalate/fluoride. However, heparin was associated with much higher (up to 50%) non-specific NAD rduction than oxalate/fluoride.
7. High levels of acetoacetate (400–1000 μmoles per litre) reduced the recovery of D-B-HOB from ovine plasma by less than 10%. This effect was negated by the inclusion of hydrazine hydrates in the reaction mixture. Perchlorate ion concentrations above 25 μmoles per litre per test dramatically inhibited the assay in ovine plasma, and therefore precipitation conditions must be carefully controlled.
8. Plasma with oxalate/fluoride as anticoagulant showed the greatest stability in storage; 24 hours at room temperature, one week at +4°C and at least one month at ?20°C.

     

The overwintering mode ofBemisia tabaci and its parasitoids in Israel

Wild and cultivated plants in the vicinity of Kibbutz Nahshon and a few additional locations in Israel were sampled for the presence ofBemisia tabaci Genna-dius (Homoptera: Aleyrodidae). The whiteflies, together with their parasites,Eretmocerus mundus andEncarsia lutea, were found to develop on numerous host species throughout the winter. Especially high levels were reached onLan-tana camara, Abutilon grandifolium andIpomoea batatas. During late winter and spring the population on these hosts declined. From April onwards the populations increased on potatoes and sunflowers.     

The hymenopterous parasites of Nitidulidae

The known information on hymenopterous parasites of Nitidulidae was consolidated in this article. Species of parasites, nitidulid hosts and, where possible, plant host of the nitidulids are provided along with citations to sources of information. Thirty-three species of hymenopterous parasites are recorded in nine different families. Since nomenclature of several of the parasites has changed, it was updated with the assistance of specialists.     

Detection of beet yellows virus in sugarbeet plants by enzyme-linked immunosorbent assay (ELISA)

Beet yellows virus can be detected in leaf extracts of infected sugarbeet plants by ELISA. The use of discs was studied and proved to be a valuable and qualitatively reliable method. Leaf material could be stored at 4^o or 22°C for at least six days without affecting the detection of this virus by ELISA. A dramatic decrease in ELISA values was found when leaf extracts were frozen.In an analysis of the distribution of virus over the plant it was found that young leaves present at the moment of infection and those which had still to develop after infection will contain virus. Symptoms produced by systemic virus invasion occur on the oldest leaves containing virus.Samenvatting Het bietevergelingsvirus kan op betrouwbare wijze met de ELISA methode in geïnfecteerde bieteplanten worden aangetoond. Een aanzienlijke vereenvoudiging van de procedure kan worden bereikt met de zogenaamde disc-method, waarbij intacte ponsstukjes in de putjes van de ELISA-plaat worden geïncubeerd. Hierbij komt voldoende virus uit de ponsstukjes voor ELISA vrij. Bladmateriaal kon op verschillende wijzen bewaard worden zonder dat de mogelijkheid om het virus aan te tonen achteruitging. Met bladextracten die ingevroren waren, werden echter slechte resultaten verkregen.In een analyse naar de verdeling van het virus over het loof bleek het virus voor te komen in de geïnoculeerde bladeren, in die bladeren die op het tijdstip van inoculatie minder dan de helft van hun uiteindelijke lengte bereikt hadden en in de bladeren die nog moesten verschijnen. De symptomen ontwikkelden zich op de oudste systemisch geïnfecteerde bladeren.     

Ollulanus tricuspis in domestic cats - prevalence and methods of post-mortem diagnosis

Ollulanus tricuspus is a parasite of the stomach of domestic cats and other animals, with a worldwide distribution. It can sometimes cause severe gastritis. Fifty-five cat stomachs were examined for O.tricuspis using three techniques. O.tricuspis was found in seven stomachs (13%) from adult cats; the number of parasites recovered from individual cats ranged from 9 to 119 (mean 54). The prevalence was higher than that previously recorded in New Zealand. Repeated dilution and sedimentation of the stomach contents and mucosal washings was found to be the most reliable detection method, being positive in all of the detected infections. Pepsin/HCI digestion of the stomach mucosa detected only 71% and punch sampling of the mucosa only 29% of these infections. This contrasts with an overseas study indicating that, in heavily infected cats, these techniques are similar in sensitivity. No macroscopic lesions were seen in the stomachs of infected cats.     

The pharmacokinetics of fenbendazole and oxfendazole in cattle

The pharmacokinetics of fenbendazole and oxfendazole in cattle are described. The pharmacokinetics of oxfendazole were not significantly different when administered orally and by intra-ruminal injection. At a dose rate of 4.5 mg/kg, administered orally, fenbendazole gave rise to mean peak concentrations in plasma of fenbendazole and oxfendazole of 0.11 and 0.13 g/ml respectively. Oral administration of oxfendazole, at 4.5 mg/kg body weight, gave rise to plasma peak concentrations of fenbendazole and oxfendazole of 0.10 and 0.20 g/ml respectively. Following intra-ruminal administration of oxfendazole, the peak concentrations were 0.11 and 0.18 g/ml respectively.     

A study of the interaction between bromocresol green dye and bovine,ovine and equine serum globulins

In human medicine it has been shown that the bromocresol green (BCG) dye-binding method for the determination of serum albumin is not entirely specific, the dye reacting also with certain human serum globulins. This causes over-estimation of albumin when reaction times are prolonged beyond 30 seconds.In the present study, serum albumin values obtained from three animal species by the immediate, i.e. less than 30 seconds, BCG reaction were compared with those by the 10-minute BCG reaction. Albumin-depleted sera were prepared using an affinity chromatography technique and their reactions and those of purified gamma globulin preparations with the dye were studied.In cattle, sheep and horses, serum albumin values obtained by the 10-minute reaction were higher than those obtained by the immediate BCG reaction, the differences being statistically significant. Purified gamma globulin did not react with the BCG dye after 10 minutes, but other globulins did. There were differences between the species in the magnitude of the reaction of their globulins with BCG dye.     

Effects of three methods of restraint an in rhesus and african green monkeys intravenous glucose tolerance testing

The intravenous administration of 0.75 gm glucose per kg and the measurement of serum glucose pretest and at 10, 20, 30, 60, 90 and 120 minutes constitute a satisfactory protocol for intravenous glucose tolerance testing of Rhesus (Macaca mulatto) and African Green (Cercopithecus aethiops) monkeys. No significant differences were noted between animals restrained with ketamine hydrochloride and those restrained with sodium pentobarbital, but the African Green males and females and the male Rhesus monkeys yielded significantly different results while being manually restrained.     

Equine hydatidosis: a new record for New Zealand

Cysts found in the liver of a horse which had never been out of New Zealand were used to infect two dogs which were slaughtered 35 days after infection. Large numbers of Echinococcus granulosus were recovered. These cestodes were compared with mature dog-sheep cestodes, using light and scanning electron microscopy and identified as the dog-horse strain of E.granulosus.