Antibody responses to avian encephalomyelitis virus vaccines when administered by different routes

Antibody responses to a commercial avian encephalomyelitis virus (AEV) vaccine administered by different routes were measured by an enzyme-linked immunosorbent assay (ELISA). Responses to single doses of vaccine administered by the ocular route to 10% of a flock were comparable with those obtained when all birds received a single dose in the drinking water. However, ocular vaccination of 5% of the flock resulted in significantly lower responses than those obtained when 10% were vaccinated. Maternal antibody was shown by the ELISA to persist in chickens from vaccinated flocks for up to 21 days after hatching. Day-old chickens with serum absorbances of < 0.3 at 492 nm, as determined by the ELISA, were shown to be susceptible to intracerebral challenge with the neurotropic Van Roekel strain of AEV.     

Plasma and extracellular volume in calves: comparison between isotopic and 'cold' techniques.

While isotopic techniques have largely superseded traditional markers for the determination of the volume of fluid compartments in vivo, they are not always convenient, especially with diarrhoeic animals. A direct comparison was therefore made in week-old calves between Evans blue and radio-iodinated serum albumin as measures of plasma volume and thiocyanate or 24sodium as measures of extracellular fluid space. The correlation coefficients were excellent (1.00, 0.96; P < 0.001) and the calves had plasma and extracellular fluid volumes of 72 +/- 2 and 438 +/- 2 ml kg-1, respectively. The latter value is, though high, typical of young animals and comparable with other data in calves.     

Evaluation of Tissue Adhesive to Contain Axonal Regeneration in Horses

Bilateral palmar and plantar digital neurectomies were completed in 10 horses (a total of 80 neurectomies) using one of three methods: (1) simple transection (guillotine method); (2) epineural capping; (3) n-butyl cyanoacrylate injected into the epineural sheath to act as a nerve sealant. Horses were regularly evaluated clinically for tenderness in and around the surgical site, as well as skin sensation at the coronary band in the heel region, during the 12-week course of the study. None of the surgical sites exhibited any signs of drainage or infection. Horses were then euthanatized, the nerve stumps were dissected from surrounding tissues, and the length and width of the tissue mass that had formed on the end of the nerve was recorded. Longitudinal and transverse sections of the nerve endings were examined histologically for numbers of proliferating axon sprouts (neuroma formation); whether the axons had penetrated the epineurium; degree of Schwann cell proliferation; degree of chronic inflammation; extent of foreign body reaction; extent of retrograde degeneration of the nerve bundles; and amount of fibro vascular proliferation. The proportion of legs exhibiting tenderness or heel sensation did not differ significantly between the three different treatments at any of the six different times they were examined. There was no difference between the three treatments in the length or width of the fibrous tissue scar on the ends of the nerves or in the number of sprouting axons from the ends of the nerves. Of 80 nerves examined, only two nerves were not confined to the epineurium. Both these nerves had been treated by simple transection. Statistically there was more chronic inflammation and foreign body reaction in the acrylic treated nerves, but no difference in Schwann cell proliferation or retrograde degeneration between the three treatments. There was slightly less fibrovascular proliferation in the transected nerves than in those subjected to epineural capping or acrylic, but the difference was not statistically significant. The use of the tissue adhesive n-butyl cyanoacrylate to prevent the continuous growth of axons after digital neurectomy seems to offer little advantage over more traditional methods of neurectomy.     

Detection of Ehrlichia risticii using an avidin-biotin-immunoperoxidase staining system

An indirect immunoperoxidase procedure using a specific anti-Ehrlichia risticii monoclonal antibody and an avidin-biotin-peroxidase staining method was used to detect E. risticii antigen in infected P388D1 murine monocytes. Several different methods of cytological fixation were used, including acetone (15 min), 95% ethanol (15 min), Bouin's fixative (5 hr), and 10% buffered neutral formalin (24 hr). The E. risticii organisms were labeled effectively and identified in cells fixed with acetone and ethanol. However, infected P388D1 cells fixed in 10% formalin or Bouin's fixative required enzymatic digestion with 1.0% trypsin for 15 min at 37 C before positive results were evident. This indirect immunoperoxidase avidin-biotin staining procedure proved to be a sensitive assay for the detection of intracellular E. risticii and may be an effective diagnostic procedure for formalin-fixed paraffin-embedded tissue.     

Duration of pituitary and adrenocortical suppression after long-term administration of anti-inflammatory doses of prednisone in dogs.

Duration and magnitude of hypothalamic-pituitary-adrenal axis suppression caused by daily oral administration of a glucocorticoid was investigated, using an anti-inflammatory dose of prednisone. Twelve healthy adult male dogs were given prednisone orally for 35 days (0.55 mg/kg of body weight, q 12 h), and a control group of 6 dogs was given gelatin capsule vehicle. Plasma cortisol (baseline and 2-hour post-ACTH administration) and plasma ACTH and cortisol (baseline and 30-minutes post corticotropin-releasing hormone [CRH] administration) concentrations were monitored biweekly during and after the 35-day treatment period. Baseline plasma ACTH and cortisol and post-ACTH plasma cortisol concentrations were significantly (P less than 0.05) reduced in treated vs control dogs after 14 days of oral prednisone administration. By day 28, baseline ACTH and cortisol concentrations remained significantly (P less than 0.05) reduced and reserve function was markedly (P less than 0.0001) reduced as evidenced by mean post-CRH ACTH, post-CRH cortisol, and post-ACTH cortisol concentrations in treated vs control dogs. Two weeks after termination of daily prednisone administration, significant difference between group means was not evident in baseline ACTH or cortisol values, post-CRH ACTH or cortisol values, or post-ACTH cortisol values, compared with values in controls. Results indicate complete hypothalamic-pituitary-adrenal axis recovery 2 weeks after oral administration of an anti-inflammatory regimen of prednisone given daily for 5 weeks.     

Plant protection problems caused by phytonematodes in Greece1

The major nematode pests of cultivated plants in Greece include root-knot nematodes Meloidogyne spp. (in almost all plants except citrus), cyst-forming nematodes Globodera rostochiensis, G. pallida (in potato), Heterodera avenae (in cereals), Ditylenchus dipsaci (in vegetables and ornamental plants) and Tylenchulus semipenetrans (in citrus). Based on observations made on samples examined at Benaki Phytopathological Institute, it was confirmed that serious infestation of new kiwi orchards with Meloidogyne spp. originated from seedlings infested at the nursery. Also the wide distribution of the nematode Tylenchulus semipenetrans in citrus orchards was mainly attributed to infested seedlings. The above observations led us, in 1985, to the decision to study the nematological fauna of nurseries in Greece.     

Transforming growth factor type beta (TGF-beta) and adipogenesis in pigs

The present study was performed on s.c. adipose tissue of fetal pigs at 35 to 110 d of gestation to examine the distribution of TGF-beta-positive cells, to localize TGF-beta immunoreactivity at the cellular level using electron microscopy (EM), and to determine the effect of TGF-beta on primary cultures of pig adipose tissue cells. Tissues for EM were fixed and embedded in LR white resin. Sections then were incubated with a polyclonal antibody specific for TGF-beta and TGF-beta was located using 20 nm colloidal gold conjugated second antibody. Tissues were fixed and embedded in paraffin for localization of TGF-beta at the light microscope (LM) level. Tissues were incubated with anti-TGF-beta followed by localization using biotinylated second antibody. Using LM, only a few cells stained positively for TGF-beta within developing blood vessels at 35 d. By 50 d, more TGF-beta-positive cells were associated with forming capillary networks. Between 70 d and 110 d, positively stained adipocytes usually were clustered around blood vessels. Cells surrounding hair follicles stained positive for TGF-beta between 90 to 110 d. Electron microscopy revealed TGF-beta labeling within fat cells. Fibroblasts and endothelial cells did not exhibit TGF-beta immunoreactivity. The addition of TGF-beta to primary cultures of s.c. adipose tissue cells from newborn pigs prevented lipid filling in fat cells. This effect was dose-dependent, with half-maximal inhibition occurring at 3 pM maximum inhibition occurred at 40 pM. These results indicate that TGF-beta may regulate angiogenic activity and lipid filling in s.c. adipose tissue of fetal pigs. Although TGF-beta was present in adipocytes and in cells associated with developing capillary networks, the physiological role of TGF-beta during early adipose tissue development is not known.