RESUMO
Epididymal spermatozoa obtained post mortem are considered a valuable source of genetic material which is often irrevocably lost. This makes these gametes constitute a key element in protection and restitution programs. The wisent (Bison bonasus, Linnaeus 1758) is a species that survived in zoos after extinction from its natural habitat. This resulted in a narrowing of the genetic pool of the whole population, which is at present derived from only 12 ancestors. Currently, wisent protection programs are aimed at preserving the genetic diversity by establishing a germplasm bank. The objective of this study was to comprehensively characterize the morphology, morphometry and functionality of wisent epididymal spermatozoa and evaluate the effectiveness of their cryopreservation in extender based on Tris buffer and chicken egg yolk. The median total number of spermatozoa obtained from one individual was 1985.0 × 106 (62.5 × 106-7452.0 × 106). These gametes were characterized by median: 40.0% (0.5-70.0%) subjective motility, 69.8% (32.5-90.0%) viability and 54.3% (10.5-83.3%) normal morphology. The sperm head had a median size of 5.0 µm (3.5-6.7 µm) width, 8.5 µm (6.4-11.3 µm) length and 36.9 µm2 (23.7-48.6 µm2) surface area. The viable population of the obtained gametes was characterized by median values 53.2% (4.5-80.3%) of intact sperm membrane, 50.8 (26.0-76.6%) of intact acrosome, 0.4% (0-98.7%) of fragmented chromatin, 5.9% (0.0-88.8%) of cells with high mitochondrial potential and 42.1% (8.3-63.7%) without lipid peroxidation. The viable population of the frozen/thawed gametes was characterized by median values: 18.4% (2.4-57.9%) of intact sperm membrane, 35.1 (11.9-56.7%) of intact acrosome, 0.07% (0-89.2%) of fragmented chromatin, 12.8% (0.0-49.7%) of cells with high mitochondrial potential and 16.3% (2.2-53.6%) without lipid peroxidation. Due to the material originating from a relatively large number of wild individuals, the research presented here contributed to the description of certain species standards for the assessment of wisent epididymal spermatozoa. The presented effect of cryopreservation on these gametes justifies the use of an extender based on Tris buffer with the addition of chicken egg yolk. The obtained effects are satisfactory from the point of view of preserving valuable genetic material and their use in ART.
Assuntos
Bison , Masculino , Animais , Trometamina , Sêmen , Espermatozoides , Criopreservação , Cromatina , GalinhasRESUMO
The aim of this study was to determine the effect of supplementation with ethyl esters of linseed oil on the fatty acid profile in hair sebum, blood serum and erythrocyte membranes in healthy dogs. The material for the study included hair and blood samples of adult beagle dogs. The experiment was performed in two periods: summer and winter. Each time it lasted 16 weeks. During the first 8 weeks, the dogs received a supplement, the amount of which was determined individually so that the ratio of α-linolenic acid (ALA) to linoleic acid (LA) together in the feed and supplement was 1:1. Hair coat and blood samples were collected on days 0, 56 and 112; i.e., before the start of supplementation, after 8 weeks of supplementation and 8 weeks after supplementation was completed. The study included a determination of the fatty acid profile with a particular emphasis on polyunsaturated fatty acids (PUFAs) and the ratio of omega-6 to omega-3 in hair sebum, blood serum and erythrocyte membranes. As a result of supplementation, a significant decrease in saturated acids and an increase in unsaturated acids was observed in hair sebum both in summer and winter and especially in omega-3 fatty acids; i.e., α-linolenic (ALA) and its derivatives eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The same relationships were observed in blood serum and in erythrocyte cell membranes in all the studied periods. Additionally, 8 weeks after the end of supplementation, the level of polyunsaturated fatty acids was still higher compared to the period before supplementation. Moreover, the supplementation resulted in a favorable decrease in the ratio of omega-6 to omega-3 acids in the tested samples, which persisted even after the withdrawal period.
RESUMO
Numerous studies have shown that improvements in the sperm and semen quality of males of many species can be achieved with appropriate dietary supplements added to feed or fodder. Particularly promising seems to be the inclusion of omega polyunsaturated fatty acids in the diets of males. Among other things, it has been shown that linseed oil ethyl esters (EELO can be an excellent source of omega 3 polyunsaturated fatty acids in animal diets. These compounds are more durable and resistant to oxidation, epoxidation and resinification processes, and do not exhibit toxic properties in living organisms. At present, there is a lack of data in the literature on the enrichment of boar diets with EELO. The purpose of this study was to analyze the effects of the addition of EELO to boar diets on the properties of sperm in fresh semen. The study was conducted during the summer on semen collected from 12 boars of the line 990. Linseed oil ethyl esters were administered in each feeding at a rate of 3.0% (45 mL each) in basal diets for each boar on a daily basis for 16 weeks. Ejaculates were collected manually by the gloved-hand technique, at one-week intervals for eight-week periods, from the eighth week onwards after the start of feeding. Eight ejaculates were collected from each boar, totaling 96 samples. The addition of EELO to the diets of boars caused an increase in sperm viability (p < 0.001), semen volume (310 mL versus 216 mL, p < 0.001) and sperm concentration (331 versus 216 million per mL, p < 0.001). Furthermore, in the experimental animals, there was a decrease in the percentage of spermatozoa exhibiting DNA fragmentation. The experimental boars also showed an increase in the percentage of gametes without apoptosis and capacitation and an increase in the percentage of viable spermatozoa not showing lipid peroxidation membranes. Consequently, EELO nutritional supplementation resulted in the improved quality of the fresh semen of boars.
RESUMO
This article presents a retrospective study on dystocia cases in bitches that were unintentionally mated and carried an unwanted pregnancy in the last 39 years. The evaluated medical records include 76 cases of difficult labour, which is 8.3% of 914 dystocia cases recorded during the period. Of these bitches, 38.2% (29/76) were 8 years, and 18.4% (14/76) were younger than 12 months. In 67/76 cases (88.2%), conservative (pharmacological and manual) obstetrical assistance proved to be unsuccessful, and caesarian section (CS) had to be performed, in contrast to the remaining recorded cases of dystocia (in which the pregnancy was intended and expected) when CS was performed significantly less often, in 71.5% (599/838) of cases. In unplanned pregnancies, 46.6% (110/236) of delivered pups were dead compared to only 26.4% (864/3273) dead pups in planned pregnancies. p value < 0.05 was considered significant. Despite the widespread availability of the spaying procedure nowadays and its safety, unplanned and unwanted pregnancies in dogs are still a concern in clinical practice. However, throughout the years investigated here, we observed an apparent decrease in the occurrence of dystocia after unintended mating, with much less recorded cases from year 2004 (71 vs. 5). Most probably, this is due to the increasing popularity of surgical castration in both females and males, and rising societal awareness of its importance, giving hope that some improvement in the welfare of dogs has already been achieved.
RESUMO
BACKGROUND: There is some controversy about the extent of changes in different sperm cell features in stored boar semen, especially regarding the potential role of the DNA fragmentation assay for assessment of sperm fertilizing ability. The aim of this study was to assess the effect of time of storage and the dynamic changes in sperm cell characteristics in normospermic boar semen stored in long-term extender, in order to determine the susceptibility to damage of particular structures of spermatozoa during cooling and storage at 17 °C for 240 h post collection. The study included five ejaculates from each of seven boars of the Polish Large White breed (n = 35 ejaculates). The sperm characteristics were assessed using a flow cytometer and a computer assisted sperm analyzer on samples at 0, 48, 96, 168 and 240 h post collection. RESULTS: The sperm chromatin structure assay (SCSA) showed a significant abrupt increase (P < 0.01) in the DNA fragmentation index (%DFI) after 48 h of semen storage with only subtle changes thereafter, not exceeding 5% on average after 240 h of storage. The use of a combination of SYBR-14/PI stains did not reveal any significant changes in the percentage of live sperm cells up to 168 h of semen storage. A significant (P < 0.01) decrease in the percentage of live spermatozoa with intact acrosomes was observed after prolonged semen storage (168 h). A significant and progressive decrease in sperm motility was recorded during the whole period of semen storage. CONCLUSIONS: Storage of boar semen extended in long-term diluent at 17 °C for 48 h initially induced a decrease in the integrity of sperm DNA. This suggests that the structure of boar sperm DNA is susceptible to damage, especially during semen extension and at the beginning of sperm storage. These findings support the opinion that the SCSA test has only a low potential for routine assessment of boar semen preserved in the liquid state and for assessment of sperm quality changes during 10 days of semen preservation. Remarkably, the integrity of acrosomes and plasma membranes remained nearly unchanged for 7 days.
