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.     

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.     

Multiple cytopenias associated with monocytic proliferation in a dog

An unusual combination of blood cytopenias and monocytic proliferation was observed in a dog. Initial hematologic findings included severe thrombocytopenia, neutropenia, mild nonregenerative anemia and apparently normal bone marrow. Subsequently, a severe persistent monocytosis developed and the bone marrow became populated with monocytes and cytophagic macrophages. Splenomegaly was due to reticuloendothelial hyperplasia and extramedultary hematopoiesis. Treatment consisted of splenectomy and azathioprine but the response was poor and the dog was euthanized. Postmortem examination revealed a hypocellular bone marrow which contained moderate numbers of monocytes and plasma cells. Neoplastic proliferation was absent in visceral organs. No definite diagnosis was established; chronic blood cell consumption, perhaps immune-mediated, may have been responsible for the extensive reticuloendothelial hyperplasia and cytophagia.     

Congenital cardiac disease in dogs

Pulmonic stenosis is caused by a malformed pulmonic valve, stricture of the right ventricular outflow tract or stricture of the pulmonary artery. English Bulldogs, Beagles, Samoyeds, Fox Terriers and Chihuahuas are predisposed. Clinical signs in severely affected dogs include exercise intolerance, stunting, dyspnea, syncope and ascites. Auscultation reveals a high-frequency, crescendo-decrescendo murmur during systole, loudest over the left side of the thorax, near the sternal cardiac border. An ECG may reveal a right-axis deviation of greater than 120 degrees, S waves in leads I, II and III, deep S waves in CV6LL, CV6LU and V10, Q waves deeper than 0.5 mv in leads II, III and AVF, and positive T waves in lead V10. Plain film LAT thoracic radiographs reveal an elevated carina, increased sternal contact of the heart, loss of the cranial cardiac waist and a widened cardiac silhouette, with normal pulmonary vasculature. A DV projection reveals an inverted "D" shape of the right ventricle and a pulmonary artery bulge. A nonselective angiocardiogram reveals poststenotic dilation of the main pulmonary artery. Treatment involves surgical correction of the stenosis.