Use of an indirect enzyme-linked immunosorbent assay (elisa) in the detection of channel catfish, Ictalurus punctatus (Rafinesque), antibodies to Edwardsiella ictaluri

Abstract. The use of an indirect enzyme-linked immunosorbent assay ( elisa ) for the detection of channel catfish antibody to Edwardsiella ictaluri is described. Changes in agglutination titre in fish immunized with Edwardsiella ictaluri heat killed whole bacterins or lipopolysaccharides were reflected by corresponding changes in elisa readings. Relatively high correlations were observed among elisa OD readings, computed elisa titres and corresponding agglutination titres.     

Serum amyloid A protein (SAA) in horses: objective measurement of the acute phase response

A sensitive and precise immunoassay for equine serum amyloid A protein (SAA) was established and used to determine, for the first time, the circulating concentration of this protein in health and disease. As in other species, equine SAA was present only at trace levels in healthy animals but behaved as an extremely sensitive and rapidly responding acute phase reactant following most forms of tissue injury, infection and inflammation, objectively reflecting the extent and activity of disease. Measurements of SAA should make a significant contribution to diagnosis and management of viral and bacterial infection in horses, and routine serial assays could provide an objective criterion for monitoring prospectively the general health of horses in training and racing.     

Host immune response to northern fowl mite: immunoblot and lectin blot identification of mite antigens

White leghorn hens were experimentally infested with northern fowl mites (Ornithonyssus sylviarum) and antibody responses to mite immunogens were monitored over 12 weeks. Mite burdens increased during the early phase of infestation and declined over the latter weeks of the study. Antigen was prepared from homogenized whole mites, which were then sonicated and extracted with non-ionic detergent. Antigen extract was fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and antibody-reactive polypeptides were identified by immunoblotting. At the start of infestation, hens had natural, pre-existing antibodies that reacted with several mite-extract components. Individual hens had different natural antibody reactivities; however, all birds had immunoglobulins reactive with extract polypeptides of 117,000, 77,000 and 36,000 molecular weight. A variety of mite extract components reacted with hen antibodies generated in response to experimental infestation. The number of antibody-reactive mite polypeptides increased through week 8 of infestation and then decreased by week 12. Fifteen polypeptides of northern fowl mite extract were reactive with antibodies developed by the majority of infested birds. These commonly reactive polypeptides had molecular weights ranging from 40,000 to 160,000. Glycoconjugates of fractionated mite extract were identified by blotting with lectins that have different carbohydrate binding specificities. Also identified were lectins that bound extract components with the same molecular weights as those moieties complexed by immunoglobulins of infested birds.     

Impact of postovulatory food deprivation on the ova transport, hormonal profiles and metabolic changes in sows.

The effect of food deprivation on ova transport, hormonal profiles and metabolic changes was studied in 20 crossbred multiparous sows during their second oestrus after weaning. To determine the time of ovulation, transrectal ultrasonographic examination was performed. The sows were divided into 2 groups, one control group (C-group), which was fed according to Swedish standards, and one experimental group (E-group). The E-group sows were deprived of food from the first morning meal after ovulation until slaughter. Blood samples were collected every second hour from about 12 h before expected ovulation in the second oestrus after weaning until slaughter and were analysed for progesterone, prostaglandin F2 alpha-metabolite, insulin, glucose, free fatty acids and triglycerides. All sows were slaughtered approximately 48 h after ovulation and the genital tract was recovered. The isthmic part of the oviduct was divided into 3 equally long segments and flushed separately with phosphate buffered saline (PBS). Uterine horns were also flushed with PBS. A significantly greater number of ova were found in the first and second part of the isthmus in the E-group (p = 0.05) while in the C-group most of the ova were found in the third part of the isthmus or the uterus (p = 0.01). The level of prostaglandin F2 alpha-metabolite was significantly higher in the E-group compared with the C-group. The concentration of progesterone increased in both groups after ovulation but there were no significant differences between the groups. The other blood parameters showed that the food-deprived sows were in a catabolic state. The 48 h period of fasting results, directly or indirectly in an delayed ova transport, which may be due to a delayed relaxation in the smooth circular muscle layer of the isthmus.     

Plasma digoxin concentration in dogs with mitral regurgitation.

Fifteen and eight mature beagles, without (normal group) and with experimental mitral regurgitation (MR group), respectively, were given 0.02 mg/kg/day digoxin powder for 10 days orally. The optimum time for sample collection after administration of digoxin was observed to be 8-18 hr and 10-22 hr in the normal and MR groups, respectively. In both groups, a stable concentration was reached after 3-5 days of treatment. No differences in plasma level were observed between sexes. The optimum concentration of digoxin was attained at an earlier stage than has been previously reported for both dogs and humans.     

Seasonal changes in serum total thyroxine, free thyroxine, and canine thyroid-stimulating hormone in clinically healthy beagles in Hokkaido.

The purpose of this study was to evaluate seasonal influences on thyroid hormone levels of healthy outdoor dogs in Hokkaido. We surveyed serum basal total thyroxine (tT4), free thyroxine (fT4), and canine thyroid-stimulating hormone (cTSH) levels, and tT4 levels after administration of TSH for a year. Basal tT4 levels decreased in January, and increased in August and September. fT4 levels increased in January and November. No significant seasonal variation was found in cTSH. tT4 levels after administration of TSH in August and November increased. These results suggested that the thyroid gland may have been activated in November. We should take seasonal variation into consideration when thyroid function is tested.     

Neurotrophins and the Thymus: Morphological Analysis of Mice Carrying a Non-functional Mutation on the Trka and Trkb Genes

Veterinary Research Communications -     

Alfalfa Phytoestrogen Content: Impact of Plant Maturity and Herbage Components

Legumes contain a range of non‐nutritional phytochemicals that may have health‐promoting effects in humans. In this study, we determined the concentrations of four phytoestrogens (coumestrol, apigenin, luteolin and quercetin) in field‐grown alfalfa (Medicago sativa L.). Differences between plants of different stages of maturity, between plant parts, and different canopy segments were assessed. The concentration of individual phytoestrogen in whole herbage varied between 15 and 225 μg g^?1 dry matter (DM) and was strongly affected by stage of maturity. Coumestrol and apigenin concentrations were highest at early vegetative stages, luteolin and quercetin at early vegetative and late flowering stages. All phytoestrogens were found in lowest concentrations at the early flowering stage (average 68 μg g^?1 DM); stage at which alfalfa is usually harvested when used as a forage source for animals. At vegetative stages, apigenin was the predominant phytoestrogen in herbage followed by coumestrol, the reverse being observed upon initiation of flowering; luteolin and quercetin were found at all stages in similarly lower concentrations. Concentrations of luteolin, quercetin and apigenin were 225, 410 and 690 % greater, respectively, in flowers than in leaves or stems; coumestrol concentration was similar between plant parts. In flowers and stems the predominant phytoestrogens were apigenin and quercetin, followed by coumestrol and luteolin. Similar concentrations (average 26 μg g^?1 DM) of each of the four phytoestrogens were found in leaves. Concentrations through the herbage canopy varied and were greatest at >60 cm from the soil surface for apigenin and coumestrol, but greatest at >60 and 0–20 cm for quercetin and at 0–20 cm for luteolin. The results suggest that if alfalfa is to be used as a source of phytoestrogens and is harvested for the production of herbal supplements or nutraceuticals, management will need to be adapted.     

Hyperadrenocorticism associated with excessive sex hormone production by an adrenocortical tumor in two dogs.

An 11-year-old spayed female Labrador Retriever and a 9-year-old castrated male miniature Poodle were evaluated because of clinical signs of hyperadrenocorticism. Cortisol testing did not support a diagnosis of hypercortisolemia in either dog; however, imaging studies revealed unilateral adrenal tumors in both dogs. Serum concentrations of 17-hydroxyprogesterone, progesterone, and estradiol were high in both dogs, and androstenedione concentrations were also high in 1 dog. It is suspected that sex hormone secretion by the adrenal tumors in these dogs resulted in clinical signs of hyperadrenocorticism. Clinical signs and hormonal abnormalities resolved in the male dog after surgical resection of the tumor. There was no improvement in clinical signs after treatment with mitotane in the female dog, which died 2 months after diagnosis. Histologic evaluation confirmed the presence of adrenocortical carcinoma in both dogs.