Early pathogenesis in chicks of infection with an enterotropic strain of infectious bronchitis virus

One-day-old specific-pathogen-free chicks were inoculated intranasally and intraocularly with infectious bronchitis virus (strain G). At days 1, 3, 5, 7, 10, and 14 postinfection, three birds were euthanatized, and the virus contents of both enteric tissues and some non-enteric tissues were assayed. Immunofluorescence and histopathological studies were also conducted. Six of 30 chicks died of nephritis between days 5-10 postinfection. Gross kidney lesions were the major pathological abnormalities. Inflammation was observed histologically in trachea, kidney, and rectum. High virus titers were found at various times in trachea, kidney, and all enteric tissues except for the jejunum. Relatively high titers of virus were still detectable at day 14 postinfection in the kidney, proventriculus, cecal tonsil, ileum, rectum, and bursa of Fabricius. Immunofluorescence staining showed viral antigens in enterocytes at the tips of villi in the ileum and rectum, and in the bursa. Viral antigens were also demonstrated in the epithelial cells of the trachea and in kidney tubules.     

Estimation of the developmental age of the bovine fetus and newborn calf

Jersey cows from several herds provided 97 fetuses and 24 calves at fixed gestational intervals between 80 and 290 days after conception. The fetuses and calves were killed, weighed and measured and, after dissection, the sizes and weights of a range of skeletal and soft tissues were recorded. Six morphological measurements emerged as most suitable for the determination of developmental age in the normal fetus. By plotting their mean values and 95 per cent tolerance limits, the rates of growth and the variability of each measurement were defined. Long bone length was the most useful single measurement for predicting the developmental age of the fetus. Brain weight, bodyweight, crown-anus length and long bone length showed curvilinear growth patterns; age prediction equations derived from these measurements are complicated to use and additional simplified formulae have been derived. The number of appendicular ossification centres also had predictive value, but it could not be used to determine fetal developmental age between 100 and 160 days gestation.     

Use of mononuclear leukocyte insulin receptor characteristics as predictors of carcass composition in heifers and steers

Total insulin specific binding (IB) and the number and affinity of insulin receptors on mononuclear leukocytes (MNL) were used to predict carcass composition of heifers and steers. Dependent variables were kidney fat, body cavity fat, s.c. fat, intermuscular fat, lean and bone. Independent variables were parameters that could be measured on the live animal, including insulin receptor characteristics, age, shrunk weight, breed and carcass s.c. rib fat thickness (SUB). All carcass fat characteristics and IB were greater for heifers than for steers, but the ability to predict either heifer or steer carcass fat characteristics was not improved by inclusion of IB in prediction equations. However, the number of low-affinity insulin receptors on MNL contributed significantly to the prediction of all heifer carcass characteristics except bone. Carcass s.c. rib fat thickness also entered the prediction equations for all heifer carcass characteristics except kidney fat. In the prediction of heifer kidney fat, the only significant independent variable was the number of low-affinity insulin receptors on MNL (R2 = .38). Carcass characteristics of steers were better predicted by SUB than were heifer carcass characteristics, and insulin receptor characteristics, when added to steer equations that contained SUB, improved R2 by .10 or less. Our results suggest that insulin receptor characteristics will be most useful in the prediction of carcass characteristics of heifers where there is a poor relationship between quantity of s.c. fat and other carcass fat depots.     

Comparative studies of the use of some injectable anaesthetic agents in birds

The effect of three injectable drugs and a combination of two of them was compared in birds. The agents were ketamine hydrochloride, xylazine hydrochloride and the steroid mixture alphaxalone-alphadolone. Ketamine and xylazine were used in combination. One hundred and fifty-four species, all maintained by major zoos in the United Kingdom, were involved in the study. The results indicate that the drugs used produced more widely differing effects in the range of species studied than have previously been reported.     

A lysosomal storage disorder in the BALB/c mouse: bone marrow transplantation

The morphological and biochemical consequences of transplanting affected bone marrow from donor BALB/c mice with a lysosomal storage disorder (BALB/c LSD) into normal recipient mice were studied. Bone marrow was removed from normal BALB/c and BALB/c LSD mice and transfused into normal BALB/c recipient mice four hours after the mice received 850 rads of irradiation. Tissues of the recipient mice were examined 240 days later. This study revealed that the defective cells that constituted the visceral lesions of BALB/c LSD could be transplanted to normal BALB/c mice by the use of bone marrow from affected BALB/c LSD homozygote; that the defective cells of BALB/c LSD proliferated and disseminated throughout the mononuclear phagocytic system of the recipient; that there were increases in cholesterol, sphingolipids, and cystine with decreases in sphingomyelinase and glucocerebrosidase activity in tissues of the recipients; and that the recipients survived substantially longer than BALB/c LSD homozygotes and their lifespan was compromised mainly by the secondary effects of irradiation. These lesions, although not as extensive as in homozygous BALB/c LSD, paralleled the lesions which develop in BALB/c LSD. Since the recipient mice were not compromised by the short life span (70 days) of the BALB/c LSD mice, they may be used to study the long-term chronic effects of these metabolic lesions.     

A practical technique for preparation of antiserum to canine factor VIII-related antigen

Identification and measurement of canine factor VIII-related antigen (VIIIR:Ag) has many clinical and research applications, including differential diagnosis of hemophilia A and von Willebrand's disease, use as a marker for specific cell types, and elucidation of the structure of this factor VIII component. We have developed a practical method for producing antibodies to canine VIIIR:Ag that uses 2% agarose filtration for purification and identifies the antigen by correlation with the elution of the peak A280 fraction in the void volume. Antisera for electroimmunoassay (EIA) can be produced in less than four weeks from simple starting materials and with commonly available laboratory equipment. This technique would be useful for either clinical veterinary or comparative research laboratories.     

Blood sampling techniques in reptiles

Disposition of the superficial blood vessels was studied in 38 species of reptiles with a view to examining their suitability for taking blood samples. It was concluded that the coccygeal veins were probably the most suitable for this purpose and the precise techniques for sampling from the different orders are described.     

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.