“Sperm mediated gene transfer” (SMGT) is based on the ability of sperm cells to bind exogenous DNA. The main objective of this study was to improve the production of transgenic pigs by SMGT. Taking into account that there is a lack of repeatability in studies of SMGT and that the mechanism of binding and internalization of exogenous DNA is a question that has not been solved, different factors involved in the production of transgenic animals by SMGT method were evaluated. Here we set out to: (1) evaluate the sperm capacity to bind exogenous DNA after DMSO treatment; (2) determine the location of the transgene–spermatozoa interaction; and (3) evaluate the efficiency of production of transgenic piglets by deep intrauterine artificial insemination (AI) with sperm incubated with DNA. The percentage of DNA binding was higher than 30% after 2 h of co-culture, but it was not affected by sperm treatment with DMSO (0.3% or 3%). The integrity of the sperm plasma membrane plays a critical role in DNA interaction, and altered plasma membranes facilitate interactions with exogenous DNA. DNA bound mainly to spermatozoa with reduced viability. DNA molecules were found to be mainly associated to the post-acrosomal region (61.9%). After deep intrauterine AI a total of 29 piglets were obtained, but none of them integrated the transgene. In conclusion, although it has been confirmed that DNA can associate with boar spermatozoa, the efficiency of producing transgenic pigs by AI was not confirmed by the present experiments, mainly due to a reduced DNA binding to functional spermatozoa. 相似文献
Forty eight 21 days old piglets were used to compare the effect of prebiotic or probiotic supplementation and ileo rectal anastomosis on the morphology of the small intestine. Half of the piglets were maintained intact and the other half was subjected to an ileo rectal anastomosis (IRA). Each group of piglets received one of the following diets: 1) basal diet (C), 2) basal diet supplemented with a Xylo-oligosaccharide (XOS), 3) basal diet supplemented with a Saccharomyces cerevisiae (SC) and 4) basal diet supplemented with XOS and SC. Villus height was greatest with XOS and with XOS + SC, only in the ileum, as compared to controls. In the duodenum, crypt width was highest in the control group, but no significant differences were found in the jejunum and ileum. The IRA piglets had longer villi in the jejunum and shorter villi in the ileum. The crypt depth was greater in the duodenum and in the ileum of IRA piglets. Villus height/crypt depth was lower in the duodenum and in the ileum, in the IRA piglets. In conclusion, the XOS, but not the SC, moderately modified the intestinal morphology. The IRA modified the intestinal villus and crypt architecture but its consequence on the absorption of nutrients needs to be investigated. 相似文献
Two maize based diets containing 2.3 g phytic P and 38 mg Zn/kg (P+) or 1.3 g phytic P and 25 mg Zn/kg (P−) were formulated. A third diet (P+/Phytase) was P+ supplemented with 3-phytase (500 FTU/kg). Each of these three basal diets, supplemented or not with 15 mg Zn/kg, was given during 20 days to weaned piglets and to 1-day-old chicks. Chicks fed the P− diets were removed from the data set because of an abnormally low feed intake.
In piglets, added Zn increased bone (P < 0.05) and plasma Zn (P < 0.01) irrespective of the basal diet (basal diet × Zn, P > 0.05). Supplementing P+ with phytase was more efficient in improving these indicators of Zn status than replacing P+ by P−. In chicks, supplemental Zn improved Zn status in a higher extent when added to P+ than to P+/Phytase (basal diet × Zn, P < 0.05). Phytase increased Zn status in a lesser extent than 15 ppm supplemental Zn. More Zn was soluble in gizzard than in stomach which corresponded to the differences in pH (4.2 vs 5.0). Phytase increased soluble Zn in stomach (P < 0.05) but not in gizzard. These results suggest a higher availability of Zn in chicks than in piglets in the absence of phytase. This may explain the higher efficacy of phytase for improved Zn availability in piglets than in chicks. 相似文献
At weaning, low feed intake level is a major factor responsible for gut disorders. However, less is known on adaptation of the antibacterial activity of pancreatic juice in response to variations in feed intake level. Moreover bovine colostrum has been demonstrated to decrease coliform counts in duodenal digesta and to improve health status of weaned piglets. Aims of the present study were to evaluate effects of feed intake level and of a bovine colostrum-supplemented diet on the pancreatic secretion of antibacterial activity in piglets at weaning. At 33 days of age, twenty-eight piglets fitted with two permanent catheters (for pancreatic juice collection and reintroduction) were weaned and allocated to one of the following three dietary treatments for 5 days: low feed allocation with a starter diet (L), gradually increased feed allocation with a starter diet (Ctrl), or gradually increased feed allocation with a starter diet supplemented with bovine colostrum (Col). In Ctrl piglets, the post-weaning secretion of antibacterial activity did not significantly differed from pre-weaning values. The pancreatic secretion of antibacterial activity in L piglets was higher than that in Ctrl piglets at day 5 (+ 300%; P < 0.05). Moreover in Col piglets the secretion of the antibacterial activity tended to increase by 55% (P = 0.08) compared to Ctrl piglets. Variations in the pancreatic secretion of antibacterial activity could constitute an adaptive phenomenon to maintain bacterial homeostasis in the upper small intestine during the early post-weaning period. 相似文献