From the Semen Collection Method to the Hatchlings: The Use of Cryopreserved Sperm from Pheasants Fed an Antioxidant-Enriched Diet.

A widely used approach to preserving genetic diversity in birds involves the cryopreservation of semen. In this process, cells are subjected to physical and chemical stresses, but not all cell species respond equally. Many studies have been published on the freezing-thawing of sperm cells from a wide variety of domestic and wild species, on issues ranging from the sperm quality to different protocols, fertilisation success rates, etc. Nevertheless, very little information is available on the common pheasant. To fill this gap, the aim of this study was to describe the pheasant semen collection method, evaluate some qualitative parameters of sperm from males fed an antioxidant-enriched diet, and to test the in vivo fertilising capacity of the cryo-preserved semen. The freezing protocol employed involved pellets thawed by the hotplate method. Dimethylacetamide was used as a cryoprotectant at a final concentration of 6%. A total of six AIs were performed at 3-4-day intervals on a total of 40 females with doses of 35 × 106 of normal live thawed sperm. Males receiving the enriched diet produce more abundant and concentrated ejaculates. Freeze-thawed sperm lost 85% of their initial mobility, and diet influenced neither sperm mobility nor viability. The enriched diet did improve the number of normal freeze-thawed cells and was associated with a lower sperm fracture incidence. Regardless of the dietary group, frozen-thawed sperm resulted in a fertility rate of 30%, with 8-9 chicks hatching for every 100 eggs incubated.

Optimization of a Protocol for the Cryopreservation of Sperm in Pellets for the Common Pheasant (Phasianus colchicus mongolicus).

The sperm of each avian species and breed have unique characteristics that render them more or less susceptible to the freezing-thawing process; therefore, a suitable cryopreservation protocol that is specific for the sperm of each type of bird is needed. In this context, little information about the common pheasant's sperm is available. Therefore, the aim of this study was to test different parameters at each step of the process of freezing into pellets and thawing to detect the least deleterious parameter settings. Sixteen different protocols were tested by studying two levels in each of the four steps (dilution, equilibration at 5 °C, final dimethylacetamide concentration, and dimethylacetamide equilibration time) comprising the freezing process. The pheasant sperm exhibited a high susceptibility to the damage caused by freezing into pellets; however, the survival of the sperm reached 29%, and the greatest recovered mobility was 22%. The mobility of the sperm was affected by the dilution and the dimethylacetamide concentration, and the viability of the sperm was affected by the equilibration at 5 °C and the dimethylacetamide equilibration. The protocols that caused the least damage to the pheasant sperm were found to be those with higher dilution rates, 10 min of equilibration at 5 °C, and 6% dimethylacetamide equilibrated for 1 or 5 min. In the present study, we individualise some applicable parameters for certain critical steps of the freezing-thawing process; however, further investigations are needed in order to improve upon and complete a suitable protocol for the cryopreservation and thawing of pheasant sperm.

Histological Characteristics, Fatty Acid Composition of Lipid Fractions, and Cholesterol Content of Semimembranosus and Triceps Brachii Muscles in Maremmana and Limousine Bovine Breeds.

This study examined the histological properties of Semimembranosus and Triceps brachii muscle in two different bovine breeds, Maremmana (Ma) (an autochthonous breed from Tuscany, Italy) and Limousine (Lm). The animals were grazed in two adjoining pastures, received the same feed supplementation, and were weighed monthly. The experimental period lasted from weaning (6 months old) to slaughter (19 months old). Muscle samples were collected immediately after slaughter, before carcass cooling. Regarding the histological properties, the number of muscle fibers (TNF), mean sarcolemma perimeter (MSP), cross-sectional area, and total sarcolemma perimeter (TSP) were determined. Samples were also analyzed for proximate composition, fatty acid profile of total lipids (TLs), phospholipids (PLs), and neutral lipids (NLs), and for total cholesterol content. Breed was a significant variation factor for the performance parameter and histological muscle fiber properties. Interestingly, despite that Ma being a less extensively genetically improved breed than Lm, it showed higher weight at slaughter (+18%) and daily weight gain (+19%). Ma also showed smaller muscle fibers than Lm and, consequently, the TSP was higher. This difference affected the lipid fraction distribution (Lm was higher in PLs and lower in NLs than Ma) and, consequently, the fatty acid composition of TLs (Lm was high in polyunsaturated fatty acids, while Ma was high in monounsaturated fatty acids). The results of this experiment highlight the importance of environmental and management conditions on the full expression of genotypic potential.

Histochemical features of the Muscovy duck small intestine during development.

We demonstrated for the first time the distribution and morphology of argyrophil and of goblet cells in the mucosa of the small intestine of the Muscovy duck during development using the Grimelius silver staining and alcian blue/periodic acid-Schiff (AB/PAS) staining technique. The argyrophil cells distribution was variable over the length of the small intestine from embryonic day 24 (24E) to post-hatching day 13 (13d). In the villi most argyrophil cells belonged to the open-type, while in the crypts they belonged to the closed-type. In the duodenum the density of argyrophil cells was highest at hatching, while in the jejunum and in the ileum the highest density value was at hatching and 13d. AB/PAS-positive goblet cells appeared on the villi and crypts of the duodenum and jejunum at 30E, and in the ileum at hatching. The density of AB/PAS-positive cells was the highest in the three segments at hatching. The AB-positive cells, compared with the PAS-positive cells, predominated in villi and crypts of the three segments, moreover the rate of AB-positive cells to PAS-positive cells significantly decreased from 30E to 9d. An increase in argyrophil and goblet cells number during the later incubation and at hatching, could indicate the small intestine in that period is being prepared to face a new diet.

Pituitary adenylate cyclase-activating peptide (PACAP) immunoreactivity distribution in the small intestine of the adult New Hampshire chicken.

We conducted a study in which we demonstrated by means of immunoperoxidase and immunofluorescence methods the presence of pituitary adenylate cyclase-activating peptide 38 (PACAP-38) immunoreactivity in the small intestine of adult New Hampshire chickens and its co-localization with VIP. In particular we describe for the first time the presence of PACAP-positive cells in the epithelium of crypts and villi. Using double immunostaining, we observed that these two peptides were widely co-localized in the nerve structures of duodenum and jejunum with the exception of the ileum, where we noticed a faint co-localization regarding the nerve fibers of the lamina propria of the villi. Furthermore, the two peptides were occasionally co-stored in the epithelial cells of the mucosa. Our findings suggest that in the chicken small intestine, PACAP can be considered, not only as a neuromodulator released by nerve elements, but also as a gut hormone secreted by endocrine cells, and it appears likely to have a role in the regulation of important intestinal physiological functions.