Follicular microvasculature and angiogenic factors in the ovaries of domestic animals

The genetic and molecular mechanisms that control the development of capillary blood vessels during follicular development are beginning to be elucidated. Ovarian follicles contain and produce angiogenic factors that may act alone or in concert to regulate the process of thecal angiogenesis. These factors are ultimately controlled by endocrine, paracrine and autocrine regulation. A recent study indicated that vascular endothelial growth factor (VEGF) plays an important role in the process of thecal angiogenesis during follicular development. We are developing a novel technology for the induction of follicular development using the technique of in vivo gene administration. Here, we summarize the recent progress of our research.     

Pathology of listerial encephalitis in chickens in Japan

Neural signs (torticollis, drowsiness) and mortality were observed in five chickens of a native chicken flock (reared for meat) that included 450 male birds on a farm that had 2300 native chickens and 1120 layers. Histologic lesions were observed in the medulla oblongata, optic lobe, cerebellum, and spinal cord of the affected birds. The lesions, which were most severe in the medulla oblongata, were massive abscesses with rarefaction (demyelination and malacia) of the parenchyma with gram-positive bacteria. The degenerative and necrotic areas were characterized by fibrin thrombosis, hemorrhages, and congestion in the blood vessels. Immunohistochemically, the bacteria positive for L. monocytogenes antigen were observed in the medulla oblongata, cerebellum, and spinal cord. Ultrastructurally, the small rod-shaped and thin-cell-walled bacteria were observed in the parenchyma of the medulla oblongata. Listeria monocytogenes (serotype 4b) was isolated from the medulla oblongata and spinal cord. The pathogenesis of listerial encephalitis in chickens was discussed.     

Pancreatic necrosis and ventricular erosion in adenovirus-associated hydropericardium syndrome of broilers

Seven 19-day-old broiler chickens affected with hydropericardium syndrome (HPS) with pancreatic necrosis and gizzard erosions were investigated pathologically and virologically. Mortality increased after 13 days of age in a flock on a broiler farm. The mortality rate of the flock reached 10% by 19 days of age. Macroscopically, the chickens had hydropericardium (the characteristic gross change of HPS), pinpoint white foci in the pancreas, and ventricular erosions. Histologically, the chickens had multifocal hepatic necrosis with intranuclear inclusions in hepatocytes, a marked increase of macrophages in the spleen and lung, mild epicardial edema, multifocal necrosis of pancreatic acinar cells with intranuclear inclusions, focal necrosis of the ventricular koilin layer, and degeneration of the ventricular glandular epithelium with intranuclear inclusions. Immunohistochemically, intranuclear inclusions in the liver, pancreas, and ventriculi were stained positively against group I avian adenovirus (GIAAV) antigens. Ultrastructurally, 67-nm diameter viral particles were present in intranuclear inclusions. Virologically, serotype 4 of GIAAV was isolated from the liver, heart, and kidney of affected chickens. The pathologic changes of the present cases differ from previous cases of HPS; therefore, the present strain of GIAAV may have different pathogenicity for chickens than the previous virus strain of HPS.     

Expression of calbindin-D9k messenger ribonucleic acid in the gastrointestinal tract of dairy cattle

The calcium demands of pregnancy and lactation are known to up-regulate expression of Calbindin-D9k (CaBP-9k) mRNA in the intestines. The gastrointestinal CaBP-9k mRNA expressions has not been studied in dairy cows, which are bound to experience several pregnancies and lactation stages. In this study, the CaBP-9k mRNA expression were examined in the gastrointestinal tract of Holstein dairy cattle by Northern blot analysis. Detectable expression of CaBP-9k mRNA was localized in the proximal portion of the small intestines. These expressions were higher at the most proximal region of the duodenum and gradually decreased distally. The duodenal CaBP-9k mRNA was detected in all dairy cattle from 0.4 to 83.4 months old, but was not detectable in foetuses. There were no significant correlations between the age and the levels of CaBP-9k mRNA expression or between the plasma 1,25-(OH)2D3 concentrations and the levels of CaBP-9k mRNA expression.     

Comparative immunohistopathology in pigs infected with highly virulent or less virulent strains of hog cholera virus

Eight pigs were inoculated subcutaneously with a highly virulent hog cholera virus (HCV) strain ALD. The infected pigs developed severe illness and became moribund on postinoculation day (PID) 7 or PID 10. Histologic lesions were characterized by severe generalized vasculitis, necrosis of lymphocytes, and encephalitis. HCV antigen was detected in crypt tonsilar epithelial cells, macrophages, and reticular endothelial cells of lymphoid tissues. Antigen localization corresponded well with histologic lesions. Five pigs were inoculated with less virulent HCV Kanagawa/74 strain and were euthanatized on PID 30. All five infected pigs recovered from the illness but became stunted. They also had a slight follicular depletion of lymphocytes, histiocytic hyperplasia, and hematopoiesis in the spleen. Less virulent HCV antigen was observed in the tonsils, kidneys, pancreas, adrenal glands, and lungs. Although antigen localization was less associated with histologic lesions, immunoreactivity was stronger than that in the pigs infected with the ALD strain of HCV. An almost complete loss of B lymphocytes was recognized in pigs infected with the ALD strain and was correlated with follicular necrosis in lymphoid tissues. Loss of B lymphocytes was not prominent in the pigs infected with Kanagawa/74 strain. The number of CD4+ and CD8+ T lymphocytes was significantly higher than that in the noninfected control pigs.     

Immunohistochemical localization of neuropeptides in bovine pancreas

The occurrence and density of distribution of nerves and endocrine cells that are immunoreactive for neuropeptides in the bovine pancreas were studied by immunohistochemistry. The six neuropeptides localized were galanin (GAL), substance P (SP), methionine-enkephalin (MENK), neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP) and vasoactive intestinal polypeptide (VIP). The exocrine pancreas was shown to have an appreciable number of GAL- and SP-immunoreactive nerve fibres but few fibres showing immunoreactivity for VIP and CGRP. Numerous MENK-, GAL-, SP-, and NPY-immunoreactive nerve fibres were seen in the endocrine portion of the pancreas. Nerve cell bodies in the intrapancreatic ganglia showed immunoreactivity for all of the neuropeptides except CGRP. Endocrine cells showing immunoreactivity for GAL and SP were observed in the large islets and islets of Langerhans, respectively. The present results indicate a characteristic distribution of neuropeptides in the bovine pancreas, which may regulate both exocrine and endocrine secretions of pancreas.     

Effects of a canine Elizabethan collar on ambulatory electrocardiogram recorded by a Holter recording system and spontaneous activities measured continuously by an accelerometer in Beagle dogs

Ambulatory electrocardiogram (ECG) has been recorded in dogs wearing a jacket to protect a Holter recording system, but the jacket was often damaged by dogs. We compared ECG recorded by a Holter recording system and spontaneous activity measured by an accelerometer in Beagle dogs with or without an Elizabethan collar. There were few significant differences in mean values (per hr) of the heart rate and the amount of spontaneous activity between dogs with or without the Elizabethan collar. Mean values (per 23 hr) of them had no significant difference between them. We concluded that the Elizabethan collar did not have any effect on ambulatory ECG and canine movements and was effective to protect the recording apparatus.     

Immunocytochemical distribution of endocrine cells in the gastrointestinal tract of the horse

Endocrine cells immunoreactive for somatostatin, gastrin, glicentin, glucagon, secretin, cholecystokinin, motilin and neurotensin were identified immunocytochemically in the gastrointestinal mucosa of the horse. Somatostatin-, glicentin- and glucagon-immunoreactive cells were very numerous in the cardiac and fundic regions of the stomach, whereas most gastrin-immunoreactive cells were confined to the pyloric region. Somatostatin-immunoreactive cells also were detected in all portions of the small intestine while gastrin-immunoreactive cells were confined exclusively to the upper portion and glicentin-immunoreactive cells were limited to the lower portions of the small intestine. Secretin-, cholecystokinin- and motilin-immunoreactive cells were observed only in the duodenum, while neurotensin-immunoreactive cells were confined primarily to the ileum. In the large intestine, somatostatin- and glicentin-immunoreactive cells were detected in the colon and rectum. The preferential location of endocrine cells provides additional information for future studies on the physiological roles of gastrointestinal peptides in the gastrointestinal tract of the horse.     

Morphological Study of Bovine Pregnant Corpus luteum Cells

The bovine pregnant corpus luteum was examined morphologically, to clarify the appearance and properties of the intramitochondrial bodies (IMB) in mitochondria of the large luteum cell (LLC). The incidence and diameter of the IMB (200–900 nm) showed a tendency to increase with the advance of pregnancy. Histochemically, the IMB reacted positively with protein, and immunohistochemically, they reacted IMB with 11 α-hydroxyprogesterone-1-11 succinyl-bovine serum albumin (P-BSA). The IMB seemed to combine with protein, and these granules did not undergo exocytosis. The cytoplasm of LLC contained another type of small electron-dense granules that were 150–200 nm in diameter, had an orifice for secretion, and reacted with relaxin. Small luteum cells (SLC) revealed crystalline-like inclusions. It is suggested that these inclusion cells participate in the synthesis of steroids.