Vitrifikacija mišjih jajnika primjenom krioprotektora etilen glikola i DMSO: patohistološka procjena.

Ten, 4- to 6-week-old BALB/c mice were randomly assigned to either control (non-vitrified, n = 5) or treatment (vitrified, n = 5) groups. Ovaries in the vitrified group were frozen sequentially by immersion into two vitrification solutions VS1: 10% ethylene glycol (EG) + 10% DMSO in holding medium (TCM-199 + 20% FBS) and VS2: 20% EG + 20% DMSO in holding medium. After thawing at 37 °C in 1.0 M sucrose, vitrified as well as non-vitrified ovaries were serially sectioned and examined histopathologically. The proportion of atretic follicles between non-vitrified and vitrified samples was significantly different (36.5 vs. 78.9%, P<0.001). No statistical difference due to vitrification was observed for the percentage of small follicles between the two experimental groups. In contrast, the rate of atresia for the growing and antral follicles in the vitrified ovaries was statistically higher than in the non-vitrified group (70.1 vs. 30.6%, P<0.001). Although many antral follicles were atretic following vitrification, sufficient follicles, especially small class, survived. Therefore, vitrification using EG and DMSO is an efficient procedure for cryopreservation of ovaries.U istraživanje su uzete dvije skupine BALB/c miševa u dobi od četiri do šest tjedana. Ukupno je bilo 10 miševa podijeljenih u pokusnu (n=5) i kontrolnu skupinu (n=5). Jajnici vitrificirane skupine bili su smrznuti sekvencionirano koristeći se uranjanjem u dvije vitrifikacijske otopine. Jedna otopina sadržavala je 10% etilen glikola (EG) + 10% DMSO u mediju za održavanje (TCIM-199 + 20% fetalnoga goveđega seruma), a druga otopina sadržavala je 20% EG + 20% DMSO u mediju za održavanje). Nakon odmrzavanja na 37 °C u 1,0 M otopini saharoze, jajnici obje skupine (vitrificirani i nevitrificirani) bili su serijski rezani i patohistološki pretraženi. Udio atretičnih folikula između vitrificirane i nevitrificirane skupine značajno se razlikovao (36,5 prema 78,9%, P<0,001). Nije ustanovljena statistički značajna razlika s obzirom na vitrifikaciju za udio malih folikula između obje skupine. Suprotno tome, postotak atrezije za rastuće i antralne folikule u vitrificiranim jajnicima bio je značajno veći nego u skupini u kojoj nije provedena vitrifikacija (70,1 prema 30,6%, P<0,001). Usprkos tome što su mnogi antralni folikuli bili atretični nakon vitrifikacije, preživjela je dovoljna količina folikula, osobito malih. Stoga se zaključuje da je vitrifikacija primjenom EG i DMSO učinkovit postupak za čuvanje smrznutih jajnika

Effect of improved diet on semen quality and scrotal circumference in the ram

The aim of this investigation was to assess the effect of improved diet above maintenance requirement on reproductive parameters, including testicular size, semen volume, sperm concentration and viability. Twelve Bakhtiary rams were allocated to two groups of six animals and were fed during a 12-week experiment period with different diets which were designed to supply maintenance and above maintenance requirements (dry matter, energy and protein). Dry matter (DM), barley and soybean meal for control and treatment groups were 1.5 kg, 8%, 0% and 2.1 kg, 25%, 17%, respectively. Semen was collected by artificial vagina every other week. Scrotal circumference, sperm concentration and total number of spermatozoa per ejaculate significantly increased in treatment group (P0.05). Likewise, there was a significant correlation between scrotal circumference and sperm production in treatment group (P<0.01, r = 0.96). It was concluded that improved dietary intake with higher energy and protein supplementation in Bakhtiary rams can improve their reproductive performance during the breeding season

The effect of betaine administration on rat sperm quality following cadmium toxicity

Background: The aim of this study is to survey the protective effects of betaine against cadmium and on sperm quality including progressive motility, sperm membrane integrity, concentration as well as testicular weight. Materials and Methods: Thirty adult male rats were allocated to the following three groups (n=10 in each group): control-saline, cadmium-saline and cadmium-betaine. Induction of testicular injury was achieved by a single injection of cadmium chloride intraperotoneally, and betaine was given orally 1 day before Cd injection and continued for 10 consecutive days. Five rats from each group were sacrificed on days 5 and 10 after Cd toxicity and sperm was taken from the epididymal tail for the evaluation. Results: Testicular weight significantly decreased by cadmium toxicity. Likewise, the percentages of sperm progressive motility, membrane integrity and concentration significantly decreased in cadmium group compared to the control rats, whereas, betaine treatment could enhance membrane integrity on day 10. Although, progressive motility increased following betaine therapy by day 10, however the difference was not statistically significant. Conclusion: Betaine may prevent cadmium-induced oxidative stress probably due to its antioxidant properties. This resulted in increase of sperm membrane integrity in addition to partial recovery of sperm progressive movement

Ghrelin promotes antioxidant enzyme activity and reduces lipid peroxidation in the rat ovary. Regul Pept 162

Antioxidant properties of ghrelin have been recently reported on various oxidative stresses in limited tissues. This study was set to examine the possible antioxidative effects of ghrelin in rat ovarian tissue. Twenty eight female adult Wistar rats were randomly allocated into control and treatment groups. Treatment group (n = 14) received 2 nmol of ghrelin as subcutaneous injection for 14 consecutive days or vehicle (physiological saline) to the control rats. The animals from both groups were equally killed on days 9 and 14 after beginning of ghrelin injection (n = 7 from each group on each day) and their ovaries were taken for later antioxidant enzyme activity assays as well as measurement of glutathione content and thiobarbituric acid reactive substances (TBARS) level. Superoxide dismutase activity was significantly higher on days 9 (P b 0.05) and 14 (P b 0.01) in the treated group compared to the control rats. By contrast, lipid peroxidation, as TBARS value, reduced significantly on both experimental days in the ghrelin-exposed animals (P b 0.05). Although, the mean activity of catalase and glutathione content was greater in the treated rats, however, the differences were not statistically significant. Slight changes occurred in glutathione peroxidase activity during the experimental period and there were no differences either on day 9 or on day 14 between groups. In conclusion, the results of the present investigation indicate for the first time the novel evidence of antioxidant properties of ghrelin in the rat ovary