Tris Extender Research Articles

Studies on the Quality and Antioxidant Status of Cryopreserved Pantja Buck Semen Supplemented with Aqueous Extract of Rosemary and Sericin in Tris Extender.

Introduction: Rosemary shrub/plant and Sericin have been documented to show antioxdative properties, however their role in improving post thaw semen quality has not been well established. Objectives: The present study was conducted to investigate the effect of Rosemary leaves extract and Sericin protein on post-thaw quality of Pantja buck semen. Methods: In the first experiment, 32 ejaculates were collected from 4 sexually mature Pantja bucks and pooled to form 8 pooled samples. The pooled samples were evaluated for seminal attributes (sperm motility, viability, morphology, plasma membrane integrity, and acrosomal integrity), status of antioxidative enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) and lipid-peroxidation (malondialdehyde) of sperm plasma membrane before dilution. In the second experiment, pooled samples were divided into three equal aliquots as group-C (Control), group-R (Rosemary), and group-S (Sericin). Aliquots of group-C were diluted in glycerolated Egg Yolk Tris (EYT) extender, whereas aliquots of group-R and group-S were additionally supplemented with 4.0% v/v Rosemary leaves extract and 0.25% w/v Sericin, respectively, and again examined for the above parameters at the post-dilution, post-equilibration, and post-thawing stages of semen freezing. Results: Significant differences (p < 0.05) were observed in the values of seminal attributes, level of enzymatic antioxidants, and lipid per-oxidation between group C and R and between group C and S at the post-thaw stage of semen freezing. However Sericin was nonsignificantly better than Rosemary in improving post-thaw semen quality. Conclusion: The results of the present study on limited semen samples in Pantja buck demonstrated that compared to the control group, both Rosemary aqueous extract (4%) and Sericin protein (0.25%) as an additive in TRIS extender, showed better cryoprotective effects resulting in improved post-thaw seminal characteristics.

Impacts of adding sucrose or trehalose to extenders with different glycerol concentrations on freezablility and fertility of buffalo bull semen

This experiment was conducted to determine the most suitable glycerol concentration (3 or 6%) and/or non-penetrating cryoprotectants (trehalose and sucrose) for the cryopreservation of buffalo semen, with the aim of enhancing the cryopreservation protocol. Semen of Egyptian buffalo were pooled and diluted with eight Tris extenders supplemented with either 6% glycerol (control group, GL6), 3% (low level, GL3), sucrose (SU, 50 mM), trehalose (TR, 50 mM), 6% glycerol together with 50 mM of sucrose (GL6SU) or 50 mM of trehalose (GL6TR), and 3% of glycerol together with 50 mM of sucrose (GL3SU) or 50 mM of trehalose (GL3TR), then frozen following the standard protocol. Findings indicated that GL3 extender resulted in the highest values of progressive motility, sperm kinematics, sperm membrane integrity, and viability of post-thawed semen (37 °C for 30 s). On the contrary, the Tris extender enriched only with SU and TR groups had the lowest values of sperm quality compared to the other groups (p < 0.05). All GL supplemented groups showed higher intact acrosome levels and lower detached acrosome and dead sperm with intact acrosome compared to those with TR and SU alone (p < 0.05). A significant increase in viable sperm was observed in the GL3, GL6, and GL3SU groups compared to the other groups (p < 0.05). The Tris extender supplemented with low glycerol (3%) significantly reduced the levels of MDA. In the in vivo fertility trial, it was shown that the pregnancy rate was higher in the GL6SU group (72%) than in the GL3SU group (68%; p > 0.05). Collectively, these results suggest that there is potential in using low glycerol (3%) as a cryoprotective agent in the medium for buffalo sperm cryopreservation without significant adverse effects compared to the addition of 6% glycerol. This study supported the sustainability of materials used in assisted reproductive technology by reducing the glycerol content in the freezing medium. Further research is needed to confirm this hypothesis.

Comparative study of Tris Turmeric, Tris Turmeric Dimethyl Sulfoxide and Tris Turmeric Ethylene glycol extenders on the cryosurvivability, sperm resistance, in- vivo fertility and antioxidant status in buffalo bull semen

The freeze-thaw process leads to structural and functional damage due to excessive accumulation of reactive oxygen species (ROS). The addition of exogenous antioxidants to sperm diluents is of great importance to overcome oxidative damage during freezing. The purpose of this study was to explore the effects of three diluents Tris Turmeric, Tris Turmeric Dimethyl Sulfoxide and Tris Turmeric Ethylene glycol on the cold survival ability of buffalo sperm. Semen was collected from five local adult male buffalo breeds. A base diluent of Tris-citric acid-fructose (TCF) was prepared, adding 20% whole egg yolk (TCFY). The Tris extender without turmeric, without DMSO, and without EG was kept as a control. Other extenders are Tris containing turmeric TT (100 ml/5 ml Tris), Tris containing turmeric dimethyl sulfoxide TTD (100 ml/5 ml Tris + 1.5% DMSO) and Tris containing turmeric and ethylene glycol TTE EG (100 ml/5 ml Tris + 1.5% EG). Semen samples were added and a pure sperm concentration of 60 × 106/ml was achieved. Frozen buffalo sperm after thawing showed significant improvements in all research parameters of the three breeding samples compared to the control. Tris Turmeric Ethylene was the type that best improved sperm survival under frozen conditions, followed by Tris Turmeric and Tris Turmeric Dimethyl Sulfoxide compared to the control. A significant decrease in sperm motility after thawing was evident as usage time increased in all expanders. There was a significant increase in total antioxidant content (TAC) and insignificant change in malondialdehyde (MDA) of the diluent used compared to the control. Conception rate (CR) was higher in Tris Turmeric Ethylene glycol (65.2%), followed by Tris Turmeric (60.3%) and Tris Turmeric Dimethyl Sulfoxide (55.9%) compared to the control (36, 7%). It can be concluded that Tris Turmeric Ethylene Glycol is considered the best agent for improving cold survival and sperm fertility, followed by Tris Turmeric and Tris Turmeric Dimethyl Sulfoxide.

Study the Different Antioxidants Effects on Serum Quality in Dogs

Dogs frequently have subfertility linked to poor semen quality (high percentage of morphologically defective spermatozoa, low rate of gradual mobility and total number of sperm .To examine the anti-oxidative effect of various concentratuons of NAC on cryopreservation in dogs sperm.The animals are divided into three groups: C0, C1 and C2. C0 receives the 0mM NAC, C1 receives 0.35mM and C2 receives 0.85mM. The physical characteristics studied of the spêrm include ejaculated volume, semen color, motility of sperms, percentage of dead sperms and abnormal sperms. After physical evaluation, the samples are diluted with Tri’s diluter. The tris diluter is egg-yolk glycerol. The samples are then cryopreserved in liquid nitrogen (-196 C) from October to December. A total of 50 samples are calculated and prepared for freezing in the following way:The motility of sperm is high for 0.35mM dose of NAC, and not very different in the control group and 0.85mM NAC group. The motility is significantly different for 0.35NAC as compared to the other two groups. The p-value (&lt; 0.05) shows the significant difference between the motility of 0.35 NAC and other groups. The sperm viability is highest in the 0.35mM NAC group (30.34). The viability of sperm is highest at 0.85mM. The membrane integrity is highest in 0.35mM NAC, while lowest in the control group( p&lt;0.05).In conclusion, dogs sperm cells can be shielded from oxidative stress by using moderate dosages of NAC supplied in Tris extender without negatively affecting dog semen's ability to freeze.

Effect of Adding Different Levels of Cerium Oxide Nanoparticles to Tris Extender on Some Parameters of Cooled Semen from Holstein Bulls

Effect of Adding Different Levels of Cerium Oxide Nanoparticles to Tris Extender on Some Parameters of Cooled Semen from Holstein Bulls

Asian elephant (Elephas maximus) seminal plasma: establishing the proteome and effect on spermatozoa when added to cryomedium.

Context The removal or supplementation of ejaculates with seminal plasma (SP) can affect cryotolerance and post-thaw survival of spermatozoa in many species. In the Asian elephant (Elephas maximus ), elucidation of the SP proteome and investigation of how it affects spermatozoa may enable improvement of cryopreservation protocols. Aims Herein, we characterise the Asian elephant SP proteome and investigate the impacts of SP on sperm cryotolerance in the presence of conspecific or heterospecific SP. Methods Proteomic analysis of Asian elephant SP was performed using mass spectrometry on nine samples from three individuals. In a separate study, SP was removed from six ejaculates and spermatozoa were resuspended in Tris extender supplemented with: no seminal plasma (NOSP), conspecific SP from ejaculates exhibiting 'good' (GSP, >60%) or mixed sperm total motility (MSP), or horse SP (HSP). Samples underwent cryopreservation, and sperm parameters were compared prior to cryopreservation and after thawing (0 and 2h). Key results Mass spectrometry identified 155 proteins from an array of families. Significant differences were observed in post-thaw sperm quality between SP treatments: high concentrations of MSP (25%, v/v) displayed greater average path and straight-line velocity immediately after thawing (P P P Conclusions and implications These preliminary findings suggest the potential of SP to enhance the cryosurvival of Asian elephant spermatozoa, with HSP showing particularly promising results compared to conspecific SP (GSP). Further research into the specific effects of Asian elephant SP proteins is warranted.

Investigation of the protective effects of different forms of selenium in freezing dog semen: Comparison of nanoparticle selenium and sodium selenite.

This study aimed to investigate the protective effects of nanoparticle selenium (SeNP) and sodium selenite (SS) on preventing oxidative stress during the freezing process of dog semen. A total of six dogs were used in the study. The ejaculate was collected from dogs three times at different times by massage method. A total of 18 ejaculates were used and each ejaculate was divided in five experimental groups. The experimental groups were designed to tris extender containing no antioxidants control, 1 μg/mL SeNP1, 2 μg/mL SeNP2, and 1 μg/mL SS1 and 2 μg/mL SS2. Extended semen were equilibrated for 1 h at 4°C, then frozen in liquid nitrogen vapour and stored in liquid nitrogen (~-196°C). After thawing, semen samples were evaluated in terms of CASA motility and kinematic parameters, spermatozoa plasma membrane integrity and viability (HE Test), spermatozoa morphology (SpermBlue) and DNA fragmentation (GoldCyto). Antioxidant enzyme activity (glutathione peroxidase; GPX, superoxide dismutase; SOD, catalase; CAT) and lipid peroxidation (malondialdehyde; MDA) were evaluated in frozen-thawed dog sperm. When the results were evaluated statistically, the progressive motility, VCL, and VAP kinematic parameters in the SeNP1 group were significantly higher than the control group after thawing (p < .05). The highest ratio of plasma membrane integrity and viable spermatozoa was observed in the SeNP1 group, but there was no statistical difference found between the groups (p > .05). Although the ratio of total morphological abnormality was observed to be lower in all groups to which different selenium forms were added, compared to the control group, no statistical difference was found. Spermatozoa tail abnormality was significantly lower in the SeNP1 group than in the control and SS2 group (p < .05). The lowest ratio of fragmented DNA was observed in the SeNP1 group, but there was no statistical difference was found between the groups (p > .05). Although there was no statistical difference between the groups in the evaluation of sperm antioxidant profile, the highest GPX, SOD and CAT values and the lowest lipid peroxidation values were obtained in the SeNP1 group. As a result, it was determined that 1 μg/mL dose of SeNP added to the tris-based extender in dog semen was beneficial on spermatological parameters, especially sperm kinematic properties and sperm morphology, and therefore nanoparticle selenium, a nanotechnology product, made a significant contribution to the freezing of dog semen.

Various Saccharides Addition of Extender Ram Efficiency to Improve Cryopreservation in Semen Etawah Crossbreed Bucks with Seminal Plasma Replacement

Phospholipase in buck seminal plasma will trigger a coagulation incident resulting in the demise of the spermatozoa during liquid semen processing using milk or egg yolk substrates diluent. A plasma replacement process is performed to avoid this possibility. The cryopreservation process leads to sperm cell damage due to the freezing process, therefore cryoprotectant agents such as saccharides are required as protective agents. This study aimed to investigate the effect of various types of saccharides on the quality of frozen semen of etawah crossbreed (PE) bucks during cryopreservation and thawing with plasma replacement using Priangan ram semen plasma. Semen was collected using an artificial vagina once a week. Fresh PE bucks semen centrifuged at 3,000 RPM for 30 min. The supernatant (seminal plasma) was removed and replaced in equal volume with Priangan ram seminal plasma. Semen was divided into four treatments: goat semen with sheep semen plasma in Tris diluent (control); control with 0.6% dextrose added (monosaccharides); control with 0.6% sucrose added (disaccharides); control with 0.6% raffinose added (trisaccharides). Semen quality including percentages of motile spermatozoa, live spermatozoa, and intact plasma membrane (IPM) were evaluated after diluted, equilibrated, and thawing, respectively. Results of this study showed that viability and IPM of monosaccharides, disaccharides, and trisaccharides were significantly (P&lt;0.05) higher than control (54.40 and 51.40; 55.00 and 53.60; 55.60 and 52.20 vs 48.40 and 52.20, respectively). The motility of disaccharides was significantly (P&lt;0.05) higher compared to control and other diluent (47.00 vs 41.00, respectively). In conclusion, different types of saccharides were effective in maintaining the quality of etawah crossbreed buck frozen semen. The addition of disaccharides seems more effective compared to monosaccharides and trisaccharides in tris extender on the quality of etawah crossbreed buck frozen semen.

Effect of Trehalose Supplementation in Egg-Yolk-Free Extender on Conventional Parameters and Gene Expression Related to Reactive Oxygen Species, Apoptosis, and Motility of Frozen Dog Spermatozoa.

The present study was conducted to evaluate the effects of trehalose supplementation in egg-yolk (EY)-free tris extender on dog spermatozoa. Pooled spermatozoa were diluted with extender 1 (EY-free tris extender supplemented with 0, 10, 15, 20, or 30 mM trehalose) and cooled (2 × 108 sperm/mL) for 1 hour at 4°C. After that, extender 2 (extender 1 containing 1 M glycerol) was added (v:v) to the diluted sperm, loaded in 0.5-mL straws (1 × 108 sperm/mL), and incubated at 4°C for 30 minutes. The sperm straws were frozen over liquid nitrogen (LN2) vapor for 20 minutes and then plunged directly into LN2. After thawing at 37°C for 25 seconds, sperm progressive motility (CASA), viability (SYBR-14/PI), apoptosis (Annexin V/PI), and reactive oxygen species (ROS; H2DCFDA/PI) were evaluated. Thereafter, the optimal concentrations of trehalose were selected, and the gene expression of BAX, BCL2, NOX5, SMOX, OGG1, and ROMO1 was evaluated after freeze-thawing. Supplementation with 20 and 30 mM trehalose significantly increased sperm progressive motility and viability compared to the control. However, trehalose had no significant effect on sperm ROS or phosphatidylserine translocation index. There were minor numerical increases and decreases in gene expression when the selected optimal concentrations of trehalose (20 and 30 mM) were compared to the control. However, there were no significant differences. We conclude that the addition of trehalose (20 and 30 mM) in EY-free extender could improve sperm motility and viability without significant effects on ROS, apoptosis, or gene expression.

SOME BIOCHEMICAL ATTRIBUTES IN SEMINAL PLASMA OF IRAQI BUFFALO BULLS AND THEIR RELATION TO THE SEMEN QUALITY

This study was undertaken to figure out the relationship between some biochemical characteristics in seminal plasma with the semen quality of Iraqi buffalo bulls (Bubalus bubalis). Semen was diluted with the Tris extender and divided based on the sperm cell's individual motility into two groups. The first group included good-semen quality (GS; n=3), whereas, the second group included poor-semen quality (PS; n=5). The differences between GS and BS groups in all seminal biochemical attributes lacked significance. A negative and significant (P≤0.05) correlation coefficient was noticed between cholesterol concentration and both principal and terminal sperms abnormality (r= -0.52), and total abnormalities (r= -0.53). A positive and significant (P≤0.05) correlation coefficient was shown between cholesterol concentration and normal sperm percentage (r= 0.53). Moreover, there was a negative and significant correlation (P≤0.05) between the triglyceride concentration and each of the percentages of individual sperm motility (r = -0.50) and of live sperm (r = -0.51). It can be concluded that some biochemical characteristics of seminal plasma can be used to predict the semen quality and fertility of Iraqi buffalo bulls.

Effect of Adding an Aqueous Extract of Aloe Vera (Aeav) to Tris Extender on some Characteristics of Goat Epididymal Spermatozoa at Different Cooling Times

Effect of Adding an Aqueous Extract of Aloe Vera (Aeav) to Tris Extender on some Characteristics of Goat Epididymal Spermatozoa at Different Cooling Times

Effect of Aqueous extract of Moringa Olivera leaves( AEMOL) on some characteristics of ram epididymal sperms under cooling conditions

The current study was conducted in the Graduate Studies Laboratory at the College of Agriculture, Al-Muthanna University, for the period from December 3, 2022 to March 9, 2023, to determine the effect of an aqueous extract of Moringa leaves on some characteristics of epididymal sperm of rams under cooling conditions for different periods. The aqueous extract of Moringa leaves was prepared and stored in the refrigerator ( 5 °C) until the time of use, the testicles were transferred to the laboratory less than an hour after the slaughter of the animal, the caudal epididymis was diluted with a Tris extender containing 0.03 concentration of aqueous extract of moringa olivera leaves.The results of this study showed a significant increase (P&lt;0.01) in the percentage of live sperm, individual motility, and the plasma membrane integrity, while the adding of AEMOL leads to a decreasing of the percentage of abnormal sperms. Increasing the cooling hours of sperm leads to a significant decrease in the percentage of live sperm, individual motility, testing the integrity of the plasma membrane with an increase in the percentage of abnormal sperm. Using the aqueous extract of Moringa leaves at different times led to a significant effect on the individual motility, percentage of the plasma membrane integrity represented by the hypo-osmotic swelling test (HOST) , While the treatment did not significantly affect the percentage of live sperm and abnormal sperm. According to the results of this study we conclude that the aqueous extract of Moringa Oleifera leaves improves the characteristics of rams epididymal sperms preserved under cooling conditions for different periods

STUDY REGARDING EFFECT OF TRISS-BASED AND CANIPLUS EXTENDERS ON SEVERAL SPERM PARAMETERS IN MEDIUM-LARGE BREED OF DOGS

"The study was carried out in different breeds of dogs owned by breeders in the city of Cluj Napoca. Mostly medium-large breeds were used and all the participating animals have been in good health during the time of acquiring the samples. The purpose of the study was to make a comparison between our own Tris-based extender and the commercial extender CANIPLUS CHILL in accordance with semen parameters with particular regards to motility, progressive motility, and length of survival of the spermatozoa. The evaluation has been done using the Computer Aided Sperm Analysis (CASA) system at the Faculty of Veterinary Medicine of Cluj-Napoca. We have found that in medium large size breeds the commercia CaniPlus extender have shown better results on the majority of the parameters over the Tris-based extender and has the ability to preserve the integrity is spermatozoa more efficiently over time compared to Tris extender. Furthermore we identified an improvement in all parameters when comparing samples on the day of collection between large breed dogs and medium-large breed dogs in both extender types. Additionally, we had results shown increase values of most parameter in tris extender when comparing it with CaniPlus extender in medium-large size breeds on the day of collection."

QUALITY OF FROZEN SHEEP SPERM USING SOYBEAN LECITHIN EXTENDER

The study aimed to study the impact of optimum levels of soybean lecithin in Tris extenders on the quality of frozen sheep sperm. The observed variable was the quality of the post-freezing spermatozoa, which covers the progressive motility, viability, abnormality and integrity of the sperm membrane. The results of the study showed that the level of lecithin in soybeans had a real influence (P&lt;0,05) on all parameters of the quality of post-freezing sheep spermatozoa. The higher the level of soybean lecithin, the higher the percentage of progressive motility, viability, integrity of sperm membranes up to 3% soybean lecithin, then a decrease of 4% soybean lecithin and decreasing sperm abnormality percentages to 3% Soybeans Lecithin. The Average percentage of progressive motility (43.79%), viability (50.85%) and membrane integrity (51.65%) of post-freezing spermatozoa treated with 3% soya bean lecithin (L3) was significantly higher (P&lt;0.05), and sperm abnormalities (13.72%) were significantly lower (P &lt;0.05) than all treatments tried.&#x0D; &#x0D; Keywords: Membrane integrity, Motility, Frozen spermatozoa Quality, Soybean lecithin

Effect of adding different levels of Artichoke (Cynara scolymus) aqueous extract to Tris extender on the Motility, Viability and some characteristics of rams semen stored at 5 °C

This study was conducted to show the effect of the aqueous extract of the leaves of Artichoke ( Cynara scolymus L.) on Tris extender and its effect on the semen traits of Awassi rams after preservation at a cooling of 5° C for different periods (0, 24, 48 and 72 hours). This study was conducted in the animal field and laboratories of the Department of Animal Production, College of Agricultural Engineering Sciences, University of Baghdad in Jadiriyah, from November 15, 2021 to April 25, 2022. Three local Awassi rams were used in this experiment. Their ages ranged from 2.5-3.0 years. The semen was collected from them by artificial vagina by one ejaculation/ram/week for 8 weeks. The semen of rams has been mixed in order to eliminate individual differences. The following tests were conducted to evaluate the semen characteristics for each motility, the percentage of viability sperm, total abnormalities, the integrity of the plasma membrane, and the acrosome's integrity. The current study showed that adding an aqueous extract from the leaves of the artichoke plant (Cynara scolymus L.) improved most of the studied traits when stored in preservation. The results indicated a highly significant (P&lt;0.01) for the treatments T1, T2 and T4, which amounted to (87.00 ± 1.86, 85.50 ± 2.02 and 86.87 ± 0.78%) compared to the control group (C), which amounted to (77.25 ± 2.11%), While the previous treatments did not differ significantly with treatment T3, which amounted to (81.87 ± 2.94%) in the percentage of live sperm during the 0 hour preservation period. At the same time, the results indicated no significant differences between treatments T1, T2, T3 and T4 compared to the control treatment and for all periods (0, 24, 48 and 72 hours) in the individual motility of the sperm. As for measuring the biochemical traits of seminal plasma, The results of AST and ALT enzymes indicated that there was no significant increase between treatments during one period, while an arithmetic superiority occurred after 72 hours of preservation for treatments T1, T2, T3 and T4 compared with the 0 hour preservation period. It is concluded from the current study that adding an aqueous extract of (Cynara Scolymus) artichoke leaves to Tris extender improved the semen quality of Awassi rams after preservation. Keywords: Ram; Semen; Antioxidants; Cooling; Artichoke; Cynara Scolymus.

Study the properties of ram semen preserved at 5°C for different periods after adding different levels of acai fruit extract.

This study was conducted to know the effect of adding an aqueous extract of Acai fruit (Euterpe oleracea Martius) to the semen extender of Awassi rams on semen Parameters after storage at cooling. This study was carried out in the animal field and laboratories of the Department of Animal Production-College of Agricultural Engineering University of Baghdad from November 15, 2021, to April 25, 2022. Three local Awassi rams were used in this experiment. Their ages were between 2.5 - 3 years, and they weighed Between 50-58 kg. Semen was collected from them using an artificial vagina to obtain one ejaculate from each ram per week. The samples were divided using Tris extender: 1.0, 1.5, 2.0, 2.5 mg Acai extract / 1 ml Tris in addition to the control group. The necessary tests were performed to evaluate the semen in terms of estimating individual motility, the percentage of sperm viability, the percentage of abnormal sperms, the integrity of the plasma membrane and the acrosome integrity. There was a significant increase (P&lt;0.05) in the percentage of sperm viability during the T3 treatment periods (48 and 72 hours) compared to the control group C for the same two periods; also there were no significant differences between all groups in the two periods (0 and 24 hours) of preservation in comparison with the control treatment C. Moreover T3 recorded a significant decrease (P &lt; 0.05) in the percentage of sperm abnormality after 24 hours of preservation when compared with the control group C in the same period, while there were no significant differences between groups T1, T2, T3 and T4 compared with the control group C in the periods 0, 48 and 72 hours. Keywords: ram semen, acai fruit, plant extracts

Addition of acai fruit extract to Tris extender and its effect on some parameters of ram semen preserved at cooling.

This study was conducted in the animal field and laboratories of the Department of Animal Production / College of Agricultural Engineering Sciences / University of Baghdad from November 15, 2021 until April 25, 2022. Three local Awassi rams were used in this experiment, their ages were between 2.5 - 3 years, and they weighed between 50-58 kg, and semen was collected from them using an artificial vagina to obtain one ejaculate from each ram weekly. The samples were divided using a Tris extender, as follows: 1.0, 1.5, 2.0, 2.5 mg Acai extract / 1 ml Tris in addition to the control group. Measurements of AST, ALT and MDA concentrations were performed in the seminal plasma. The results of the current study showed that it had no significant effect on the concentration of MDA, AST and ALT in seminal plasma for the two periods ( 0 and 72 hours) of preservation for treatments T1, T2, T3 and T4, although the concentration of ALT enzyme and the concentration of MDA in the seminal plasma of T3 treatment were mathematically lower than the rest of the treatments in the two periods (0 and 72 hours), while there was an arithmetic decrease in the concentration of AST enzyme for treatment T4 over the rest of the treatments in the period (0 hours) and treatment T1 over the rest of the treatments after (72 hours) of preservation. Keywords: Acai fruit extract, semen, ram, preservation, 5 °C.

Africanized honeybee (Apis mellifera) semen freezing using Tris-based and Collins extenders.

This study is aimed at evaluating the effect of different extenders on the cryopreservation of semen from Africanized honeybees (A. mellifera). Semen from honeybee drones from 10 different colonies was obtained by endophallus exposure technique and immediately evaluated for motility, viability using fluorescent probes, functional membrane integrity using the water test, and morphology. Samples from each colony were divided in three aliquots and subjected to a dilution ratio of 12:1 (diluent: semen) using Tris, Tris + egg yolk (Tris+EY), and Collins extender. Samples were cryopreserved and stored in liquid nitrogen for one week and then rewarmed and reevaluated. Immediate dilution provoked no significant effect on sperm motility and functional membrane integrity, regardless of the extender used; however, the greatest values (P < 0.05) for normal sperm morphology were found at the use of isolate Tris (69.3 ± 1.9%). After thawing, there were no significant differences among extenders with relation to the preservation of sperm motility, viability, and functional membrane integrity, but the Tris extender provided the highest post-thawing values (P < 0.05) for sperm normal morphology (49.2 ± 4.9%) while the Collins extender provoked the highest amounts (P < 0.05) of curled tail defects (67.5 ± 3.2%). Moreover, the Tris was the only extender at preserving the proportion of normal sperm after thawing similar to what was verified for fresh samples. In summary, we suggest the use of a Tris-based extender for the cryopreservation of Africanized honeybee semen.

Effect of Water Extract of Maringa Olivera Leaves on some Characteristics of Goats Epididymal Sperms Under Cooling Conditions

The current study was conducted at the postgraduate laboratories, Agriculture College, Al-Muthanna University, during the period between January 5, 2021 to May 29, 2022, to determine the effect of the water extract of Moringa olivera leaves on some characteristics of goats epididymal sperms under cooling conditions for different periods. The water extract of Moringa, Olivera leaves was prepared and stored in the refrigerator until the time of uses, the epididymal sperms were obtained from the slaughtered goats testes in the local abattoir of Al-Muthanna Governorate, and transferred immediately to the laboratory to collect the epididymal sperms from the tail of epididymis and extended by Tris extender contains different concentrations of water extract of Maringa Olivera (0.02 and 0.03)%. The results showed that the water extract of Moringa led to a significant increase (P≤0.05) on the individual motility of sperms and the hyper osmosis swelling test (HOST), with significantly decreased on percentage of abnormal and dead sperms. The increase of the time of cooling leads to a significant decrease on the individual motility of sperms and the HOST, with a significant increase on the abnormal and dead sperm. The interaction between the level of water extract of Moringa olivera and the cooling periods, there was a significant effect on the individual motility of sperms, while the interaction did not significantly affect to the abnormal and dead sperm as well as the HOST. It was concluded that the water extract of Moringa Olivera leaves improved the characteristics of goats epididymal sperms under cooling conditions.

Enhancing post-thaw quality of ram epididymal sperm by supplementation of rutin in cryopreservation extender

The purpose of this study was to examine the effect of different rutin concentrations on rams epididymal sperm. A local slaughterhouse provided 50 pair of testes from 25 rams. The testes were sent to the lab at room temperature. Spermatozoa were extracted by suspending portions of cauda epididymis in tris solution. Ram sperm was cryopreserved (in liquid nitrogen) in a tris extender containing rutin at 0, 0.5, 0.75, 1, and 1.25 mM. Rutin showed superior sperm total and progressive motility, beat cross frequency, straight line velocity, velocity average pathway and membrane integrity values at 0.75 and 1 mM. The morphology of the sperm and the superoxide dismutase levels did not significantly change with different treatments. Moreover, rutin at 0.75 and 1 mM was also shown to have the highest level of mitochondrial activity. The results showed ATP, total antioxidant capacity, and glutathione peroxidase levels were significantly greater in the rutin 0.75 and 1 mM groups (P < 0.05). Rutin at 0.75 and 1 mM levels had the lowest reactive oxygen species concentrations. Rutin at 0.75 and 1 mM substantially increased the proportion of viable sperm (P < 0.05). The lowest amount of apoptosis was observed in 0.75 and 1 mM rutin. Rutin at 0.75 and 1 mM yielded the least significant percentage of dead sperm. It may be inferred that adding 0.75 and 1 mM to the sperm extender can enhance the quality of the epididymal sperm in rams after the cryopreservation process.