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.

     

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.     

Age of the earliest african anthropoids

The earliest fossil record of African anthropoid primates (monkeys and apes) comes from the Jebel Qatrani Formation in the Fayum depression of Egypt. Reevaluation of both geologic and faunal evidence indicates that this formation was deposited in the early part of the Oligocene Epoch, more than 31 million years ago, earlier than previous estimates. The great antiquity of the fossil higher primates from Egypt accords well with their primitive morphology compared with later Old World higher primates. Thus, the anthropoid primates and hystricomorph rodents from Fayum are also considerably older than the earliest higher primates and rodents from South America.