Protein microanalysis of animal tears

Sub-microlitre volumes of normal koala, mouse, dog, rat and cat tears were fractionated using size exclusion-high performance liquid chromatography (SE - HPLC), giving reproducible profiles which were different for each species. Microlitre volumes of tears were also fractionated using sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS - PAGE), resulting in good separation of individual tear proteins with a species specific distribution. Tears from koalas with conjunctivitis and mice with keratitis were similarly examined and showed mostly quantitative changes. These simple, rapid techniques gave reproducible results and, in contrast to conventional separation techniques, used easily obtainable volumes (as little as 0.75 microl) of tears. Their expansion could allow isola tion, identification and quantitation of individual tear components, enabling effective investigation of changes occurring in disease.     

Endometrial Adenocarcinoma and Mucometra in a 6-year-old Alaska Malamute Dog

A 6-year-old female Alaska Malamute dog was presented for evaluation of abdominal enlargement referred by a local veterinarian. On the history, the owner complained of chronic abdominal enlargement initiated more than 4 months ago, reduced appetite, occasional vomiting and general dullness. He also complained of greenish mucous intermittent vaginal discharge starting 10 days ago. The bitch was chronically treated with medroxiprogesterone acetate. A laparatomy was performed and fluid in the abdomen was found and aspirated during the surgery. Also a very fluid-filled distended uterus and a mass in the distal part of the left uterine horn were found. The mass was encapsulated by the omentum, but areas of necrosis and calcification were identified. Histopathological diagnosis was endometrial adenocarcinoma.     

Oestradiol Induced Inhibition of Neuroendocrine Marker Expression in Leydig Cells of Adult Rats

The objectives of this work were to determine the changes in the expression of neuroendocrine markers in Leydig cell by oestradiol treatment, and to determine whether testosterone is able to recover partially the effects of hormonal suppression induced by oestradiol. Adult male rats were injected daily with either 50 microg of oestradiol or oestradiol plus testosterone propionate (25 mg every 3 days) for 15 days. The animals were sacrificed and testicles were dissected and processed by routine histological protocols. FSH and LH serum levels were determined by radioimmunoassay. The visualization of antigens was achieved by the streptavidin-peroxidase immunohistochemical method. Antibodies against chromogranin A (CrA), S-100 protein (S-100), P substance (PS), synaptofisin (SYN), neurofilament protein (NF), gliofibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were used. The mean LH and FSH serum concentrations were consistently suppressed with hormonal treatments. Intermediate filaments (NF and GFAP) showed no difference in their expression. The expression of S-100, NSE and SYN was significantly lower in both hormone-treated groups. In oestradiol-treated rats, the immunoreactivity of CrA and SP decreased significantly but was restored after testosterone supplementation. Although the nature and functions of many of these substances in Leydig cells remain unknown, these results are consistent with the hypothesis that the expression of some neuroendocrine markers is hormonally controlled.     

Malignant pheochromocytoma presenting as a bradyarrhythmia in a dog

Objective: To describe a case of atrioventricular block and syncope secondary to systemic hypertension in a dog with malignant pheochromocytoma. Case Summary: A 13‐year‐old spayed female mixed‐breed dog presented following an acute onset of collapse. The dog was hypertensive and had paradoxical high‐grade second‐degree atrioventricular block with concurrent syncope. Initial emergent therapy included a nitroprusside infusion for the systemic hypertension and vagolytic therapy for the bradyarrhythmia. A left adrenal mass was seen on abdominal ultrasound and was further characterized by MRI. Following medical stabilization with phenoxybenzamine and propantheline, a left adrenalectomy was performed. Histological diagnosis was consistent with a malignant pheochromocytoma. Following tumor removal there has been no further hypertension, atrioventricular block or syncope. Unique information provided: This is the first documented case of a malignant pheochromocytoma presenting as syncope secondary to hypertension induced atrioventricular block.     

Phosphorus cycling and soil P fractions in Douglas-fir and red alder stands

Nitrogen-fixing species can dramatically increase soil acidity and organic matter content, and potentially alter biogeochemical P dynamics. We compared ecosystem P cycling in adjacent stands of N₂-fixing red alder (Alnus rubra Bong.) and non-fixing Douglas-fir (Pseudotsuga menziesii Franco) in order to determine whether P-cycling rates within stands were related to soil P forms as measured by sequential P fractionation. Above-ground annual P uptake was 61% greater in the red alder stand, although soil available P, as measured by Bray (NH₄F–HCl) extraction, was only 10% of that found in the Douglas-fir stand. Total ecosystem P in the alder stand was only 69% of that found in the Douglas-fir stand, and could indicate a pre-establishment difference between stands. However, the percentage of total soil P released by Bray or NaOH extraction was also lower in the alder stand, which suggests that differences in total P alone did not control the patterns observed in P fractions. Concentrations of inorganic P sorbed to Fe and Al minerals and contained in Fe minerals and apatite were greater under Douglas-fir, while organic P was slightly greater under red alder. While fluxes of P in litterfall, uptake and resorption were 94, 60 and 292% higher in the alder stand, soil extractable fractions meant to represent available P were lower under alder. Static measures of available P do not appear to adequately reflect P supply, and the development of techniques to assess P turnover is needed to better understand cycling and plant availability of P.     

Relationship of blood corticosterone, immunoglobulin and haematological values in young crocodiles (Crocodylus porosus) to water temperature, clutch of origin and body weight

Objective To examine whether sub-optimal temperature induced stress and immunosuppression in farmed saltwater crocodile (Crocodylus porosus) hatchlings. Design A clinico-pathological study. Animals A total of 140 hatchlings were used. Procedure Body weight and length, plasma corticosterone and immunoglobulin concentrations and total and differential white blood cell counts were measured in 140 hatchlings from five clutches divided between five water temperature treatment groups. Initially all groups were housed at 32°C for 10 weeks, then two groups (L, LC) were changed to low temperature (28°C) and two groups (H, HC) to high temperature (36°C), while one group (C) remained at 32°C. The LC and HC groups were maintained at these temperatures for 10 days, after which the water temperature of both groups was returned to 32°C. Blood samples were collected twice (at 6 and 9 weeks of age) before the initial temperature change, and at 10 days and 4 weeks after the initial temperature change (at 11.5 and 14 weeks of age). Results Except for an increase in plasma corticosterone in the HC group and a decrease in the L group when the temperature change was first introduced, changes in plasma corticosterone were not significant. There were no significant changes in immunoglobulin concentrations. There were, however, significant decreases in the total white cell and lymphocyte counts in the LC group after the temperature was decreased to 28°C, and an increase in these counts after water temperature was returned to 32°C. Clutch of origin had significant effects on body weight and length gains, and there were negative relationships between body weight and corticosterone concentrations and between body weight and immunoglobulin concentrations. Conclusions As haematological changes indicative of stress were not associated with significant changes in serum corticosterone, immunosuppression in young crocodiles may be independent of the hypothalamic-pituitary-adrenal cortical axis.