A detailed study of the ontogenesis of deer stomach has not been undertaken to date, and our aim was to sequence several histological phenomena that occur during the ontogenesis of one of the gastric compartments, the rumen. Histomorphometric and immunohistochemical analyses were carried out on 50 embryos and fetuses of deer from the initial stages of prenatal life until birth. For the purposes of testing, the animals were divided into five experimental groups: group I, 1.4 -3.6 cm crown-rump length, 30-60 days, 1-25% of gestation; group II, 4.5- in its configuration the stratum basale, the lamina propria and the submucosa. From the pluripotential blastemic tissue at 60 days we witnessed the histodifferentiation of the primitive tunica muscularis, composed of two layers of myoblasts with a defined arrangement. It was also from the pluripotential blastemic tissue, at 97 days, that the lamina propria and the submucosa were differentiated. The serosa showed continuity in growth as well as differentiation, already detected in the undifferentiated outline phase. The tegumentary mucosa of deer rumen was shown without secretory capacity in the initial embryonic phases; neutral mucopolysaccharides appeared from 67 days. The presence of neuroendocrine cells (non-neuronal enolase) in the ruminal wall of deer during development was not detected until 97 days. The glial cells were detected at 142 days for glial fibrillary acidic protein and at 67 days for vimentin. The immunodetection of neuropeptides vasointestinal peptide and neuropeptide Y progressively increased with gestation period, starting from 97 days. In terms of the structure of the rumen of the primitive gastric tube, our observations revealed that the deer is less precocious than small and large domestic ruminants. Thus its secretory capacity, detected by the presence of neutral mucopolysaccharides, and its neuroendocrine nature, determined by the presence of positive non-neuronal enolase cells, were evident in more advanced stages of prenatal development than those detected in the sheep, goat and cow.

Iberian red deer need to be conserved for their economic role and for their genetic importance as an important component of the ecosystem. Modifications currently being made to traditional management systems require a better understanding of the structure, function and development of their alimentary system. Here we describe a histomorphometric and immunohistochemical analysis of the stomach of 25 red deer embryos and fetuses from 30 days of gestation until birth (235 days). Differentiation of the reticular compartment from the primitive gastric tube begins at 67 days, forming a three-layered structure: epithelium, pluripotential blastemal tissue and serosa.

The reduced lifespan of cryopreserved spermatozoa in the mare reproductive tract has been attributed to both capacitative and apoptotic changes. However, there is a lack of studies investigating both phenomena simultaneously. In order to improve our knowledge in this particular point, we studied in raw and frozen-thawed samples apoptotic and capacitative markers using a wide battery of test based in flow cytometry. Apoptotic markers evaluated were caspase 3 activity, externalization of phosphatidylserine (PS), and mitochondrial membrane potential. Markers of changes resembling capacitation were membrane fluidity, tyrosine phosphorylation, and intracellular sodium. Conventional and computational flow cytometry using nonlinear dimensionally reduction techniques (t-distributed stochastic neighbor embedding (t-SNE)) and automatic classification of cellular expression by nonlinear stochastic embedding (ACCENSE) were used. Most of the changes induced by cryopreservation were apoptotic, with increase in caspase 3 activation (P < 0.01), PS translocation to the outer membrane (P < 0.001), loss of mitochondrial membrane potential (P < 0.05), and increase in intracellular Na+ (P < 0.01). Average values of markers of capacitative changes were not affected by cryopreservation; however, the analysis of the phenotype of individual spermatozoa using computational flow cytometry revealed the presence of subpopulations of spermatozoa experiencing capacitative changes. For the first time advanced computational techniques were applied to the analysis of spermatozoa, and these techniques were able to disclose relevant information of the ejaculate that remained hidden using conventional flow cytometry.

The red deer is an important study species because of its value in the national economy and because it provides a wealth of genetic material. To date, there has been little research into the prenatal development of the stomach of ruminants, and none of the red deer. We therefore performed a histological evaluation of the ontogenesis of the omasum in the red deer. Histomorphometric and immunohistochemical analyses were carried out on 50 embryos and fetuses of deer from the initial stages of prenatal life until birth. For test purposes, the animals were divided into five experimental groups: Group I (1.4 -3.6 cm crown-rump length, CRL; 30 -60 days, 1-25% of gestation); Group II (4.5 -7.2 cm CRL; 67-90 days, 25 -35% of gestation); Group III (8-19 cm CRL; 97-135 days, 35-50% of gestation);Group IV (21 -33 cm CRL; 142 -191 days, 50 -70% of gestation); and Group V (36 -40 cm CRL; 205-235 days, 75 -100% of gestation). At 67 embryonic days, the omasum wall was differentiated, and comprised three layers: the epithelial layer, pluripotential blastemic tissue and serosa. The stratification of the epithelial layer was accompanied by changes in its structure, with the appearance of four laminae of different sizes; in order of appearance these were: primary at 67 days, secondary at 90 days, tertiary at 97 days and quaternary at 135 days. At around mid-gestation, lateral evaginations were formed from the stratum basale of the primary and secondary smaller laminae. These were the primitive corneum papillae. From 205 days, the corneum papillae were present in all four sizes of laminae.The histodifferentiation of the lamina propia-submucosa, tunica muscularis and serosa showed patterns of development similar to those reported for the rumen and reticulum of red deer. The omasum of red deer during prenatal life, especially from 67 days of gestation, was shown to be an active structure with full secretory capacity. Its histological development, its secretory capacity (detected by the presence of neutral mucopolysaccharides) and its neuroendocrine nature (detected by the presence of positive non-neuronal enolase cells and the neuropeptides vasoactive intestinal peptide and neuropeptide Y) were parallel to the development of the rumen and the reticulum. However, its prenatal development was later than that of the omasum in sheep, goat and cow.

Ultrastructural and immunohistochemical techniques were used to study the second cell type in sheep embryo pineal glands. Thirty-two embryos were studied from day 54 of development through birth. Specimens were arranged in four age groups, defined in terms of the most relevant histological features: Group 1 (54-67 days of prenatal development), Group 2 (71-92 days), Group 3 (98-113 days), and Group 4 (118-150 days). At 98 days, a second cell type was observed which differed from pinealoblasts and showed uniform ultrastructural characteristics similar to those of astrocytes in the central nervous system. Ultrastructural homogeneity was not matched by the results of histochemical and immunohistochemical analysis: while all Type II cells stained positive to phosphotungstic acid hematoxylin, only 50% expressed glial fibrillary acidic protein. In the course of ovine intrauterine development, the vascular affinity of this second cell population, composed of glial-like or astrocytic cells at varying stages of maturity, leads to the formation of a limiting pineal barrier. This barrier may constitute the morphological expression of a hypothetical functional involvement in the exchange of substances between blood and pineal parenchyma.

No abstract

Summary An experimental model was designed to characterize lesions in the lung of lambs inoculated with bovine respiratory syncytial virus (BRSV). Twenty‐five Merino lambs of both sexes, with a live weight of 17 ± 3 kg, received an intratracheal inoculation of 20 ml saline solution containing 1.26 times 106 TCID50 BRSV (strain NMK‐7) per ml. Lambs were slaughtered 1, 3, 7, 11 and 15 days postinoculation (DPI), and samples were taken for analysis using light‐microscopic and immunohistochemical techniques. The results reflected the effect of the virus on airway epithelia. The presence of BRSV in ciliated bronchial and bronchiolar epithelial cells gave rise to cytopathological changes, including loss of cilia and cell necrosis; these changes might be expected to decrease the efficiency of mucociliary clearance, favouring the development of secondary bacterial bronchopneumonia. These results suggest a reduction in BRSV tropism for alveolar epithelia compared to bronchial and bronchiolar epithelia. Light‐microscopic analysis revealed a narrowing of alveolar and airway lumina and a considerable interstitial inflammatory reaction.

Taken together, these results indicate that treatment with melatonin in goat kids in situations of stress due to premature weaning could play an important role in the improvement of histo-physiological function of the pineal gland.