Antioxidant effect of lemon balm (Melissa officinalis) and mate tea (Ilex paraguensys) on quality, lipid peroxidation and DNA oxidation of cryopreserved boar epididymal spermatozoa.

In this study, we investigated the protective ability of the addition of two antioxidant herb extracts, mate tea and lemon balm, on boar epididymal frozen-thawed spermatozoa quality. Testes from mature boars were collected at local slaughterhouse, and sperm samples from epididymis were recovered by flushing. Spermatozoa were cryopreserved in lactose-egg yolk buffer supplemented with various concentrations of lemon balm and mate tea (0, 2.5, 5 and 10 g l(-1) ) using the straw-freezing procedure. Motion parameters, acrosome and plasma membrane integrity, lipoperoxidation levels and DNA oxidative damage (8-hydroxy-2'-deoxyguanosine base lesion) were evaluated. There were no differences among experimental groups with regard to motility characteristics, viability, acrosome and plasma membrane integrity; however, the highest concentration of lemon balm produced significant (P < 0.05) improvement in curvilinear trajectory, straightness and amplitude of lateral head displacement after thawing. The supplementation of freezing extender with mate tea and lemon balm reduced sperm lipid membrane peroxidation, and only mate tea protected DNA against oxidative damage during cryopreservation at 120 min post-thawing (P < 0.05). Mate tea experimental extender at concentration of 10 g l(-1) showed the lowest percentage of sperm oxidised DNA and malondialdehyde generation; thus, mate tea is a potential candidate such as antioxidant compound on boar sperm cryopreservation medium.

Cryopreservation of epididymal stallion sperm.

Any event that makes semen collection or mating impossible, such as death, castration, or injury, may terminate a stallion's breeding career. Fortunately, stallion sperm which are capable of fertilization can be harvested from the epididymis, and frozen for future use. However, the fertility of frozen-thawed epididymal sperm has been found to be lower than that of ejaculated sperm. Therefore, this study aimed to optimize the fertility of frozen epididymal stallion sperm by investigating the effects of different cryoprotectants and freezing protocols on sperm quality. Dimethylformamide was tested alone or combination with pasteurized egg yolk as substitute of fresh egg yolk. In addition, the effect of the pre-freeze stabilization on sperm quality was analyzed. Heterospermic samples obtained from stallion epididymis were collected and cryopreserved in lactose-egg-yolk extender or in the same extender with varying content of cryoprotectant and content of egg yolk, stabilized and no-stabilized. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. No improvement was observed on the replacement of fresh yolk by pasteurized egg yolk, whereas the results suggest that dimethylformamide is a cryoprotectant suitable for cryopreservation of equine epididymal semen, even better than glycerol. In addition, we found that the stabilization before freezing on epididymal stallion sperm, can improve sperm quality parameters.

Determination of ovulation time in sows based on skin temperature and genital electrical resistance changes.

Different physical and physiological parameters may be used to determine ovulation time in sows. In the present study, we analysed the ear and vulvar skin temperature fluctuations, and the changes in genital electrical resistance, at a distance of 4, 8 and 12 cm from the vulva during oestrus in order to predict the time of ovulation. Multiparous sows were checked by transrectal real-time ultrasonography and luteinising hormone (LH) plasma concentration was determined. Temperature was measured using a thermoprecision infrared thermometer, and the electrical resistance was measured with a commercial resistance probe. All measurements were carried out every 12 hours from one day after the weaning to three days after oestrus onset. Skin temperature showed significant difference around periovulatory period. The electrical resistance at 4 cm from the vulva showed marked changes during oestrus, which were different from those described at 8 and 12 cm from the vulva. At 12 hours before ovulation time, skin temperature decreased significantly, and negative correlation (P<0.05) was found between vulvar skin temperature and vaginal resistance. There was no relationship between skin temperature, electrical resistance and LH plasma concentration. The measurement of several physiological traits may provide more accurate predictions of the moment of ovulation.

Crystallisation pattern of vestibular mucus and its relation to vestibular electrical resistance in cycling sow.

Changes in the genital mucus around the oestrus are used by different diagnostic methods to determine optimal fertilisation time. In the current study, the authors evaluated the different arborisation patterns found in vestibular mucus, and also established its relationship with vestibular resistance changes during oestrus. Thirty multiparous sows were checked by transrectal ultrasonography to determine ovulation time every 12 hours. Vestibular resistance was measured with a commercial resistance probe, and vestibular mucus ferning was also evaluated every 12 hours during the oestrus. Significant changes (P < 0.05) in vestibular resistance were detected, registering high variation among individuals. Maximum resistance data was reached between 12 and 24 hours after ovulation time in 83 per cent of the sows. Crystallisation samples were classified into three different patterns according to the fern-like crystal degree. Arborisation peak occurred from 48 to 36 hours before the moment of ovulation, when vestibular resistance values increased gradually. In the optimal insemination moment, vestibular resistance increased significantly (P < 0.05) and vestibular mucus showed a low crystallisation pattern (P < 0.05). Combining several methods to measure genital mucus changes may predict the ovulation time and the best insemination moment.

Dimethylformamide is not better than glycerol for cryopreservation of boar semen.

To improve the boar sperm cryopreservation process, the influence of the sugar (lactose, trehalose) source and the cryoprotectant [glycerol, dimethylformamide (DMF)] on the success of freezing was investigated. Sperm samples were frozen in one of six extenders: lactose plus 3% glycerol (LG); lactose plus 1.5% glycerol and 1.5% DMF (LGD); lactose plus 3% DMF (LD); trehalose plus 3% glycerol (TG); trehalose plus 1.5% glycerol and 1.5% DMF (TGD); trehalose plus 3% DMF (TD). Effects on motility, viability, acrosome integrity and hypoosmotic test (HOST) were measured. The results showed that extender containing 3% glycerol retained the highest motility percentages. In regard to viability and acrosome integrity, all extenders yielded similar rates except for the decreasing values of TD. Endosmosis was diminished in TD and LD at 2 h (P = 0.0018), as compared with the others. The results of the study demonstrated that the use of DMF as a cryoprotectant adversely affected boar sperm quality after cryopreservation.