The effect of different concentrations of laminarin on the quality of cryopreserved ram semen.

BACKGROUND: Increasingly, sheep breeders are using artificial insemination to produce lambs, so finding methods that preserve ram sperm can be useful. OBJECTIVE: To determine the protective effects of different concentrations of laminarin on ram sperm motility, viability, abnormalities, membrane, and DNA integrity, superoxide dismutase enzyme (SOD) activity, and malondialdehyde (MDA) production after freeze-thawing. MATERIALS AND METHODS: The ejaculates of four rams were collected and stored at 35 degree C. Semen samples were diluted with a tris-base extender containing 100, 200, 400, and 800 ug/mL of laminarin and a control extender containing no laminarin, then frozen in liquid nitrogen after 4 h in the refrigerator. RESULTS: In the treatment of frozen-thawed spermatozoa with 800 ug/mL laminarin, motility, viability, membrane integrity, and DNA integrity were significantly higher than in the control. In spermatozoa that were exposed to 800 ug/mL laminarin after thawing, MDA production was significantly lower than in the control group. The percentage of abnormal spermatozoa in 800 ug/mL laminarin was significantly lower than that in the control. CONCLUSION: The addition of 800 ug/mL laminarin to the freezing extender increases motility, viability, SOD activity, and plasma membrane integrity, while reducing abnormality and MDA production in freeze-thawed ram semen. https://doi.org/10.54680/fr24110110812.

Role of oleic acid and trehalose on frozen-thawed ram semen.

BACKGROUND: When sperm are cryopreserved, reactive oxygen species (ROS) are formed that are detrimental to the sperm. OBJECTIVE: To evaluate the effects of oleic acid and trehalose added to ram semen extender on sperm parameters, lipid peroxidation (MDA), and superoxide dismutase (SOD) enzyme levels of spermatozoa following the freeze/thawing processes. MATERIALS AND METHODS: Ejaculates were collected from four rams and pooled at 35 degree C. Pooled ejaculates were diluted with oleic acid at 0 mM and trehalose at 0 mM (O0 T0) as the control. The Tris-based extender was supplemented with either 0.5 (O0.5) or 1 (O1) mM of oleic acid or 25 (T25) or 50 (T50) mM of trehalose alone, and in combination [0.5 mM oleic acid + 25 mM trehalose (O0.5 T25), 0.5 mM oleic acid + 50 mM trehalose (O0.5 T50), 1 mM oleic acid + 25 mM trehalose (O1 T25) and 1 mM oleic acid + 50 mM trehalose (O1 T50)]. The semen was frozen by the traditional liquid nitrogen vapour method and stored at -196C in the liquid nitrogen tank. RESULTS: Semen extender containing O1T25 significantly improved the total motility, when compared with other treatment groups (P<0.05), except for O1 T50. O1 T50 had a higher viability rate than any other treatment. The addition of O1 T25 and O1 T50 increased DNA and membrane integrity of spermatozoa post-thawing compared to other treatments (P<0.05). The level of MDA was significantly (P<0.05) lower in extenders supplemented with O1, O0.5 T25, O0.5 T50, O1 T25 and O1T50 compared to the other treatment groups. In addition, SOD levels were higher in groups treated with O1 T25 and O1 T50 than the other treatment groups (P<0.05). CONCLUSION: The addition of a combination of oleic acid and trehalose concentrations to Tris-based extender improved the quality of ram semen post-thawing. Doi.org/10.54680/fr23610110712.

[Congenital glaucoma: Intraocular pressure and visual prognosis after trabeculectomy and functional rehabilitation for amblyopia]. / Glaucome congénital : pronostic pressionnel et visuel après trabéculectomie et rééducation de la part fonctionnelle de l'amblyopie.

The goal of our work is to analyze the clinical aspects, study the change in intraocular pressure and functional prognosis after trabeculectomy and amblyopia treatment of children followed for congenital glaucoma. The present study is a retrospective study including 86 patients (143 eyes) followed over a period from March 2009 to September 2015. The median age at diagnosis was 6 months. The mean intraocular pressure was 25±5mmHg preoperatively and 11.6±4mmHg at the conclusion. The average initial cup was 0.5. Trabeculectomy was performed in all cases. Twelve eyes were operated twice. After a follow-up of 4 years, normalization of the intraocular pressure was obtained in 35% after the first surgery, in 44% on mono or dual therapy, in 10% after a surgical revision. Cycloplegic refraction was performed; myopia was found in 55% of cases, the mean was -6.5 diopters. 20% of children were hyperopic with a mean of 2.5D. A mean 2D of astigmatism was found in 60 eyes (42%). Anisometropia was present in 10 children. Corrected visual acuity was quantified in 37% of children. The mean was 2/10±3 initially and 4/10 at the conclusion. It was≥4/10 in 41% of cases and≤1/10 in 56%. Unilateral amblyopia was found in 40% of the children. Seventy percent of the patients had strabismus. This study highlights the need for prolonged treatment of amblyopia in congenital glaucoma to achieve the best possible visual recovery.

Combination of oocyte and zygote selection by brilliant cresyl blue (BCB) test enhanced prediction of developmental potential to the blastocyst in cattle.

The cumulus oocyte complexes (COCs) were obtained from local abattoir. After aspiration, the COCs were allotted into four treatments to evaluation of brilliant cresyl blue (BCB) test. Control treatment (C): oocytes were cultured directly (without exposure to BCB) after recovery in in vitro production (IVP) process. Oocyte treatment (OBCB): immediately after aspiration, COCs were incubated in modified Dulbecco's phosphate-buffered saline (mDPBS) supplemented with 26µM of BCB for 90min and classified into two classes: oocytes with blue cytoplasm coloration (OBCB+: more competent oocytes) and oocytes without blue cytoplasm coloration (OBCB-: low competent oocytes). Directly after classification, the oocytes were maintained undisrupted in the IVP process. Zygote treatment (ZBCB): After oocyte collection, maturation and fertilization, zygotes were stained with BCB for 10min and categorized into three ways, according to whether they were highly stained (ZBCB++: low competent zygotes), moderately stained (ZBCB+: moderate competent zygotes) and unstained (ZBCB-: more competent zygotes). Directly after classification, the zygotes were maintained undisrupted in the culture process. Oocyte and zygote treatments (OBCB/ZBCB): COCs were stained with BCB after recovery and classified into two classes (OBCB+ and OBCB-). After fertilization, the zygotes produced from OBCB+ and OBCB- oocytes were further stained with BCB for 10min and categorized six ways (OBCB+/ZBCB++, OBCB+/ZBCB+, OBCB+/ZBCB-, OBCB-/ZBCB++, OBCB-/ZBCB+ and OBCB-/ZBCB-). Directly after classification, the zygotes were maintained undisrupted in the culture process. The selection rate produced from OBCB treatment (OBCB+; 54.3%) was greater (P<0.05) than ZBCB treatment (ZBCB-; 44.3%). In addition, the selection rate produced from double application (combination of oocyte and zygote selection) of BCB test (OBCB+/ZBCB-: 28.8%) was less (P<0.01) than single application of BCB test (ZBCB-: 44.3%or OBCB+: 54.3%). The percentage of blastocyst production from OBCB+ oocytes (35.7%) and ZBCB- zygotes (36.6%) were greater (P<0.05) than that from C oocytes (25.7%), OBCB- oocytes (16.5%), ZBCB++ (13.5%) and ZBCB+ zygotes (21.3%). However, there were no significant differences (P>0.05) in the percentages of blastocyst production between OBCB+ oocytes (35.7%) and ZBCB- zygotes (36.6%). The proportion of blastocyst production from double application of BCB test (OBCB+/ZBCB-: 48.0%) was greater (P<0.05) than that from single application of BCB test (OBCB+: 35.7% or ZBCB-: 36.6%). In conclusion, current results confirmed that combination of oocyte and zygote selection by BCB test enhanced the efficiency of selecting for high quality embryos, compared to the single BCB test.