Significant Difference In Sperm Motility Research Articles

Trehalose, Glycerol and Dimethyl Sulfoxide Supplementation for Cryopreservation Media of Honey Bee Drone Semen

The cryogenic storage of honey bee drone (Apis mellifera) semen is a rapidly developing practice, driven by the need to preserve bee colonies and the decline in gamete quality following cryopreservation. Advances in cryobiology, coupled with procedural refinements and the development of new methods for cryopreservation of male gametes at different temperatures, provide promising results for improving drone semen preservation. Innovative strategies involve the incorporation of additives within preservation media to enhance important spermatozoa parameters. Our study aimed to assess the effectiveness of cryoprotectants (CPAs), namely trehalose, glycerol, and dimethyl sulfoxide (DMSO), on the motion characteristics of honey bee drone sperm cells during cryopreservation and subsequent thawing. Using Sperm Class Analyzer (SCA) and Computer-Assisted Sperm Analysis (CASA), we evaluated the impact of these supplements on sperm motility and kinetic parameters. Our findings indicated significant differences in sperm motility (non-progressiveness and progressiveness) and kinetics (curvilinear velocity and linearity) across treatments with trehalose, DMSO, and glycerol. Trehalose supplementation retained significantly higher percentages of sperm motility and kinetic parameters post-cryopreservation compared to DMSO and glycerol. Nevertheless, despite the utilization of new protocols, media, and supplementation, the results obtained are often unsatisfactory. The research provides valuable information for enhancing the efficacy of cryopreservation media to optimize the successful outcome of honey bee drone spermatozoa storage.

Performance evaluation of sperm concentration, motility, and morphological analysis for GSA-810 series of sperm quality analysis system.

The performance evaluation of each computer-assisted sperm analysis (CASA) system may provide a basis for the interpretation of clinical results and further improvement of the CASA system. The accuracy of the GSA-810 CASA system was evaluated by detecting latex bead quality control products. The precision of sperm concentration, morphology, and percentages of progressively motile sperm (PR) were evaluated by coefficient of variation (CV). Three samples with sperm concentration of about 100 × 106 /mL were diluted to evaluate the linear range. The detection values of latex beads were within the range of target values. The CVs of sperm concentration and PR were significantly and negatively correlated with sperm concentration (r = -0.561, p = 0.001) and PR value (r = -0.621, p < 0.001), respectively. The R2 values of the linear range of sperm concentration were ≥0.99. There was no significant difference in sperm motility and PR within 1-10 min at 36.5°C ± 0.5°C. The coincidence rates of sperm morphology and sperm head morphology for 36 semen samples analyzed by the GSA-810 system and manual method were 99.40% and 99.67%, respectively. The CVs of the percentage of sperm with abnormal morphology and percentage of sperm with abnormal head morphology were less than 5%. The GSA-810 system can accurately analyze normal semen samples, but the repeatability of the results is poor for oligozoospermia and asthenozoospermia samples. The future CASA system for analyzing sperm morphology should focus on recognizing the middle and tail segments of a spermatozoon.

Antioxidant Activity of Fermented Red Bean Extract on Sperm Quality of Mice Exposed to Cigarette Smoke

Smoking has a negative effect on fertilization as it decreases sperm quality. The current research aimed to investigate the effect of fermented red bean (Phaseolus vulgaris L.) extract on sperm motility, viability, and plasma membrane integrity of white mice (Mus musculus) exposed to cigarette smoke. The red beans were subjected to a 36-hour fermentation process using Rhizopus spp. Then, methanol was extracted by maceration method for 24 hours until maceration was obtained. In this study, 25 male white mice aged 3 months were randomly divided into 5 groups of 5 mice. Group C (negative control) was given 0.5 mL of carboxymethyl cellulose natrium (CMC Na) 1% solution orally without unfiltered cigarette smoke exposure, and group C+ (positive control) was given 0.5 mL of CMC Na 1% solution orally and unfiltered cigarette smoke exposure. Treatment groups T1, T2, and T3 were orally given fermented red bean extract at doses of 26 mg/kg Body weight (BW), 52 mg/kg BW, and 104 mg/kg BW, respectively, and then were exposed to unfiltered cigarette smoke. For 36 days, treatment groups (except the negative control) were subjected to the inhalation of smoke from an unfiltered cigarette containing a nicotine dose of 2.2 mg. The exposure period lasted for 20 minutes each day. Each group was put into a cigarette smoke-exposing box. The sperm motility (observing the forward movement of spermatozoa), the sperm viability (examining the color of the sperm head), and the sperm plasma membrane integrity (observing the tail shape using the hypoosmotic swelling test) were then evaluated. The findings indicated significant differences in sperm motility, viability, and plasma membrane integrity of each group with positive control. A dose of 104 mg/kg BW of fermented red bean extract had the best potential to maintain sperm motility (70%), viability (82.13%), and plasma membrane integrity (61.93%) of mice exposed to unfiltered cigarette smoke.

Comparison of Pregnancy Outcomes in Thyroid Antibody-Positive Women with Different Fertilization Methods

<i>Objective</i> To compare the pregnancy outcomes of thyroid antibody (ATA) positive women treated by different fertilization methods during in vitro fertilization-embryo transfer. Methods A total of 204 female patients with normal thyroid function and positive ATA in the reproductive Medicine Center of the First Hospital of Shanxi Medical University from January 2020 to January 2022 were selected. According to different fertilization methods, they were divided into IVF group and ICSI group, including 115 IVF cycles and 89 ICSI cycles. The general data, clinical indicators after ovulation induction, fertilization, embryo status, and pregnancy outcome of the two groups were compared. <i>Results</i> There were no significant differences in age, BMI, bE₂, FSH, FT₃, FT₄, AFC, Gn dosage, Gn stimulation time, E₂ on HCG day, P on HCG day, intrauterine thickness on HCG day, number of retrieved oocytes and average number of embryos transferred between the two groups (P>0.05). The fertilization rate of the ICSI group was higher than that of IVF group within the group, while the high-quality embryo rate was significantly lower than that of the IVF group, with statistical significance (P<0.05). There were significant differences in sperm motility and sperm density between the two groups (P<0.05), and there was no significant difference between sperm volume and normal morphology rate (P>0.05). There were no significant differences in embryo implantation rate, biochemical pregnancy rate, clinical pregnancy rate, early abortion rate and live birth rate between the two groups (P>0.05). Logistic regression analysis showed that the method of assisted pregnancy and semen quality were not independent factors affecting high-quality embryo (P>0.05). In IVF group, TGAb was negatively correlated with fertilization rate (r=-0.202, P<0.05); and in ICSI group, TPOAb was negatively correlated with high quality embryo rate (r=-0.349, P<0.05). <i>Conclusion</i> Thyroid autoimmunity (TAI) in infertile women undergoing assisted reproductive technology (ART), the semen quality should be considered as the priority criteria for ATA. ICSI should not be recommended as routine fertilization.

Safety Evaluation of bioactive Sub-Fraction of Lawsonia inermis Linn leaves in Male Wistar Rats

This study evaluated the safety of bioactive labelled sub-fraction of Lawsonia inermis in male Wistar rats. Twenty male rats ‎were used for this study of five rats per group. Three groups were administered methanol bioactive sub-fraction of Lawsonia ‎inermis at 2.5, 5 and 10 mg/kg for fourteen days while control received distilled water orally. Physiological weight increased in ‎all treatment groups. The group fed with 10 mg/kg had the highest weight gain compared to other treatment groups and the ‎control. Relative organ weight of the heart, liver, kidneys, testes and pancreas did not increase (p&gt;0.05) in all the treatment ‎groups except the pancreas in the group fed with 5 mg/kg that increased significantly (p&lt;0.05). The sperm count in rats fed at ‎dose rate of 10 mg/kg increased significantly (p&lt;0.01) compared to other treatment groups. There were no significant ‎differences in sperm motility, volume and live/dead ratio in the treatment groups compared to control. Haematological ‎parameters such as, PCV, RBC, WBC and haemoglobin concentration were not significantly different compared to control. ‎The AST increased significantly (p&lt;0.01) while urea and creatinine decreased significantly (p&lt;0.001) in group fed with 2.5 ‎mg/kg. 5 mg/kg showed significant reduction (p&lt;0.05) in total protein, urea and ALP while ALT increased significantly. Serum ‎chloride ion at the dose rate of 2.5 mg/kg decreased significantly (p&lt;0.05) when compared with other treatment groups. ‎Sodium ion (Na+) decreased significantly (p&lt;0.01), while Bicarbonates (HCO3-) increased significantly in the 5 mg/kg group. It ‎was concluded that the bioactive sub-fraction of Lawsonia. inermis was safe at the administered dosages.‎

Effects of preconception exposure to phthalates on mouse sperm capacitation parameters.

Phthalates have been linked to adverse male reproductive health, including poor sperm quality and embryo quality as well as a longer time to pregnancy (months of unprotected intercourse before conception occurs). The present study aimed to evaluate the effect of preconception exposure to two ubiquitous phthalate chemicals, di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DBP), and their mixture on sperm function, fertilization, and embryo development in mice. Adult male C57BL/6Jmice aged 8-9weeks were exposed to di(2-ethylhexyl) phthalate, di-n-butyl phthalate, or their mixture (di-n-butyl phthalate+di(2-ethylhexyl) phthalate) at 2.5mg/kg/day or vehicle for 40days (equivalent to one spermatogenic cycle) via surgically implanted osmotic pumps. Caudal epididymal spermatozoa were extracted and analyzed for motility using computer-assisted sperm analyses. Sperm phosphorylation of protein kinase A substrates and tyrosine phosphorylation, markers of early and late capacitation events, respectively, were analyzed by Western blots. In vitro fertilization was used to evaluate the sperm fertilizing capacity. While the study did not reveal any significant differences in sperm motility and fertilization potential, abnormal sperm morphology was observed in all phthalate exposures, particularly in the phthalate mixture group. In addition, the study revealed significant differences in sperm concentration between control and exposed groups. Moreover, protein phosphorylation of protein kinase A substrates was decreased in the di(2-ethylhexyl) phthalate and mixture exposure groups, while no significant changes in protein tyrosine phosphorylation were observed in any of the groups. Assessment of the reproductive functionality did not reveal significant effects on in vitro fertilization and early embryo development rates but showed wide variability in the phthalate mixture group. Our findings suggest that preconception phthalate exposure affects sperm numbers and phosphorylation of protein kinase A substrates involved in capacitation. Future research is warranted to examine the associations between phthalate exposure and capacitation in human spermatozoa.

The effect of individual selection on sperm motility in tropical abalone Haliotis squamata

High-quality broodstock is required to support successful cultivation activities of the tropical abalone. This study aimed to find out the structure of the sperm under the influence of individual selection. A total of 50 eggs per batch were measured in egg diameter, calculated by the degree of fertilization and hatching rate. Sperm collection is carried out from the natural spawning of male abalone from the wild (F0) and the selected offspring from individual Fillial-1, F2, and F3 selections. Samples of sperm were collected by artificial spawning and analyzed using the Sperm Class Analyzer® CASA System. The results showed that the wild abalone (F0) had the highest hatching rate but was not significantly different from other derivatives (P &gt; 0.05). The average spermatozoa was 2.39 million cells/m, with the highest spermatozoa (F0) of 7.110 million cells/ml. The most increased sperm motility and rapid velocity were in the wild (89.78%; 34.00%) and the lowest of F2 (22.00; 4.00%). A significant difference in sperm motility and rapid velocity were found between F0 and other treatment. The consequences of individual selection will aid in studying reproductive factors and sperm motility, which are most likely crucial in tropical abalone fertilization.

Polymorphisms in the HSF2, LRRC6, MEIG1 and PTIP genes correlate with sperm motility in idiopathic infertility.

The aim of this study was to investigate the association of 24 functionally important single nucleotide polymorphisms (SNPs) with male infertility. In this cross-sectional study, we genotyped 24 functionally important single nucleotide polymorphisms in 24 infertility candidate genes in 500 oligo-/astheno-/oligoastheno-/normo-zoospermic infertile men with idiopathic infertility. Sequenom iPlex gold assay was used for genotyping. Sperm count and motility were compared between prevalent genotypes at each test locus. We did not observe any significant difference in the average sperm count between the alternate genotypes for the loci in the KLK3, LRRC6, MEIG1, HSF2, ESR2 and PTIP genes. However, we observed a significant difference in sperm motility between the alternate genotypes for the loci in the LRRC6, MEIG1, HSF2 and PTIP genes. Polymorphisms in the LRRC6 (rs200321595), MEIG1 (rs150031795), HSF2 (rs143986686) and PTIP (rs61752013) genes show association with sperm motility.

Semen characteristics and second successful artificial insemination of Asian elephant (Elephas maximus) in Thailand.

Background and Aim:As the number of wild Asian elephants (Elephas maximus) continues to decline, maintaining healthy populations under human care is vital. Male fertility assessment is essential for understanding the reproductive status, which can help to uncover underlying problems and improve the rate of pregnancy success. The objectives of this study in Asian elephants were as follows: (1) To investigate the semen characteristics; (2) to compare the relative seminal vesicle size and semen characteristics; (3) to compare the semen characteristics between good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates; and (4) to investigate the pregnancy success rate after artificial insemination (AI) with combined chilled and frozen semen.Materials and Methods:In total, 153 ejaculates were collected by manual rectal stimulation from 25 bulls. The volume, pH, sperm concentration, progressive motility, viability, morphology, and membrane integrity were investigated in each ejaculate. Assessment of accessory sex glands was conducted using transrectal ultrasonography to compare the relative seminal vesicle size and semen characteristics, and the bulls were divided into two groups according to the size of the ampulla (<7 or ≥7 cm2). For the comparison of good and poor-motile ejaculates and semen characteristics, the samples were divided into two groups: Good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates. Semen ejaculates for AI were collected from three bulls. The estrous cycles of four females were monitored using an enzyme immunoassay. Seven AI attempts were conducted using frozen and/or chilled semen by endoscopic visualization. AI was repeated 1 day before the luteinizing hormone surge, on the day of the surge, and 1 day after the surge. Pregnancy was confirmed by monitoring the serum progesterone profile and the abdomen and mammary glands changes.Results:From 153 ejaculates, the mean±standard error values of progressive motility, semen volume, sperm concentration, pH, and viability were 40.18%±2.28%, 40.94±3.86 mL, 1,205.58±62.26×106 sperm/mL, 7.50±0.10, and 56.17%±1.96%, respectively. Comparing ampulla size and semen characteristics revealed that the bulls with ampullae of ≥7 cm2 yielded significantly larger volume ejaculates. However, there were no significant differences in sperm motility and concentration. The comparison of semen characteristics between good- and poor-motile ejaculates revealed that the former had significantly higher pH, viability, normal acrosomes, intact membranes, and normal head and tail morphology but often had a significantly lower volume and sperm concentration. From seven AI attempts in four females, one female had a confirmed pregnancy (14.3% pregnancy rate), and delivered a healthy live female baby weighing 128 kg at 21 months and 12 days of gestation. The baby is now 3 years old and in a healthy condition, with normally developing growth and behavior.Conclusion:The semen characteristics of Asian elephants can be used as the baseline reference for further applications. The ampullae size indicates semen quantity but not quality. Our success in producing an elephant calf from AI using frozen and chilled semen demonstrated that AI can be used as an alternative approach for the breeding management of Asian elephants. However, the semen of Asian elephants is of poor quality, especially in terms of membrane integrity; thus, the improvement in semen quality through intensive and careful management of elephant health and fertility remains a challenge for the future. Furthermore, a sperm bank should be established to develop sperm cryopreservation, which will be invaluable for improving the genetic diversity of the Asian elephant.

Interferon γ, IL-17, and IL-1β impair sperm motility and viability and induce sperm apoptosis.

Urogenital inflammation is a known cause of male infertility. Increased levels of inflammatory cytokines, leukocyte counts and oxidative stress are highly detrimental for sperm quality thus compromising male fertility. Although cytokines affect sperm by recruiting and activating leukocytes consequently inducing tissue inflammation and oxidative stress, scarce to absent data have been reported about the putative direct effects of inflammatory cytokines on spermatozoa. Herein, we analyzed whether IFNγ, IL-17A, IL-1β, and IL-8 can alter human sperm motility and viability per se. Fractions of viable and motile spermatozoa from normospermic healthy donors were in vitro incubated with recombinant human IFNγ, IL-17A, IL-1β or IL-8 and sperm ROS production, motility, viability and apoptosis were analyzed. Sperm exposed to different concentrations of IFNγ, IL-17A and IL-1β, or a combination of them, for either 1 or 3h showed significantly increased levels of mitochondrial ROS production and reduced motility and viability with respect to sperm incubated with vehicle. Moreover, the exposure to IFNγ, IL-17A and IL-1β resulted in significantly higher levels of early and/or late apoptotic and/or necrotic spermatozoa. Interestingly, no significant differences in sperm motility, viability and apoptosis were observed in sperm incubated with the concentrations of IL-8 analyzed, for either 1 or 3h, with respect to sperm incubated with vehicle. In conclusion, our results indicate that IFNγ, IL-17A and IL-1β per se impair sperm motility and decreases viability by triggering increased mitochondrial ROS production and inducing sperm apoptosis. Our results suggest that screening inflammatory cytokines in semen would be an additional helpful tool for the diagnostic workup of male infertility.

Ultra-low temperature cryopreservation and −80 °C storage of sperm from normal-male and pseudo-male Siniperca chuatsi

The purpose of this study was to evaluate the effects of ultra-low temperature cryopreservation and − 80 °C storage of sperm from normal-male and pseudo-male mandarin fish (Siniperca chuatsi). In this study, sperm motility, plasma membrane integrity, mitochondrial membrane integrity, DNA integrity, fertilization, hatching rate and offspring growth were evaluated. After thawing, there was no significant difference in sperm motility and motility parameters of stripped sperm and testicular sperm of normal-males and pseudo-males. The motility of frozen sperm thawed in a 37 °C water bath was the highest, and there was no significant difference in the motility of frozen sperm of different tubes (n = 30) after thawing. The motility of frozen sperm of normal-males and pseudo-males were more than 60%, and the DNA fragmentation were less than 10%. The integrity of mitochondria and plasma membrane were higher than 84% and 75% respectively. There were no significant differences in plasma membrane, mitochondrial membrane integrity and DNA fragmentation between fresh sperm of normal-males and pseudo-males, nor between frozen sperm. The fertilization and hatching rate of normal-male frozen sperm were 65% and 85%, and values of pseudo-male frozen sperm were 57% and 83%, respectively. There was no significant difference in the body weight and length of offspring of frozen sperm between the normal-males and pseudo-males. The sperm motility, motility parameters and mitochondrial membrane integrity of the frozen sperm of normal-males and pseudo-males did not significantly decrease after −80 °C storage for 1, 3 and 5 days. The fertilization and hatching rate of normal-male and pseudo-male frozen sperm were still higher than 50% and 75%. Frozen sperm can successfully reproduce offspring after 5 days of −80 °C storage. The results indicated that testicular sperm of normal-male and pseudo-male S. chuatsi were successfully cryopreserved by ultra-low temperature cryopreservation, and the research of −80 °C storage showed that frozen sperm can be successfully stored for a short time at −80 °C which can provide a possibility for the dry ice transportation of frozen sperm.

Improving Sperm Cryopreservation With Type III Antifreeze Protein: Proteomic Profiling of Cynomolgus Macaque (Macaca fascicularis) Sperm.

Antifreeze protein III (AFP III) is used for the cryopreservation of germ cells in various animal species. However, the exact mechanism of its cryoprotection is largely unknown at the molecular level. In this study, we investigated the motility, acrosomal integrity, and mitochondrial membrane potential (MMP), as well as proteomic change, of cynomolgus macaque sperm after cryopreservation. Sperm motility, acrosomal integrity, and MMP were lower after cryopreservation (p < 0.001), but significant differences in sperm motility and MMP were observed between the AFP-treated sperm sample (Cryo+AFP) and the non-treated sample (Cryo–AFP) (p < 0.01). A total of 141 and 32 differentially expressed proteins were, respectively, identified in cynomolgus macaque sperm cryopreserved without and with 0.1 μg/ml AFP III compared with fresh sperm. These proteins were mainly involved in the mitochondrial production of reactive oxygen species (ROS), glutathione (GSH) synthesis, and cell apoptosis. The addition of AFP III in the sperm freezing medium resulted in significant stabilization of cellular molecular functions and/or biological processes in sperm, as illustrated by the extent of proteomic changes after freezing and thawing. According to the proteomic change of differentially expressed proteins, we hypothesized a novel molecular mechanism for cryoprotection that AFP III may reduce the release of cytochrome c and thereby reduce sperm apoptosis by modulating the production of ROS in mitochondria. The molecular mechanism that AFP III acts with sperm proteins for cellular protection against cryoinjuries needs further study.

Comparative analysis of various step-dilution techniques on the quality of frozen Limousin bull semen.

Background and Aim:Indonesia has two National Artificial Insemination centers and 17 Regional Artificial Insemination Centers. The frozen semen production techniques differed between the centers, including the type of diluent and semen dilution technique. The aim of the research was to compare the quality of frozen Limousin bull semen diluted using different techniques.Materials and Methods:Semen was collected from three sexually mature Limousin bulls using an artificial vagina. Immediately after collection, the semen was evaluated macroscopically and microscopically. Semen that had >70% motile sperm and <20% sperm abnormality was divided into three tubes and diluted with skim milk-egg yolk (SMEY) using three different dilution techniques: One-step dilution (100% SMEY with 8% glycerol) at room temperature ([RT] 20°C until 25°C) two-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol after 1 h stored at 5°C); and three-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol added twice at 1 h and 1.5 h after being stored at 5°C). The diluted semen was loaded into 0.25 mL mini straws, equilibrated, and frozen using a freezing machine. Sperm motility, viability, membranes, DNA integrity, and concentrations of malondialdehyde (MDA) and aspartate aminotransferase (AST) enzymes were evaluated after thawing.Results:The results showed that there were no significant differences in sperm motility and DNA integrity between dilutions (p>0.05). However, sperm viability and membrane intactness of one-step dilutions were higher than those of three-step dilutions. The concentrations of MDA and AST enzymes of sperm in one-step dilutions were lower than those of three-step dilutions (p<0.05).Conclusion:It was concluded that the one-step-dilution technique was better than three-step dilution for cryopreservation of Limousin bull semen.

Porcine seminal protein-I and II mRNA expression in boar spermatozoa is significantly correlated with fertility

In recent years, genomic and proteomic biomarkers have been identified for the diagnosis of male fertility to overcome the limitations of conventional semen analysis. Owing to the limited genes available so far, the single gene approach is commonly adopted for analyzing the phenotype of interest. However, the single-gene approach is less effective than multiple-gene strategies for diagnosing a desirable phenotype. Herein, we investigate the ability of two fertility-related genomic markers (porcine seminal protein (PSP)-I and PSP-II) in spermatozoa to predict boar litter size in addition to conventional semen parameters. First, we examined different semen parameters (motility, motion kinematics, and capacitation status) and gene expression in high- and low-litter size boar spermatozoa. Then, we evaluated the correlation of these parameters with the fertility of 21 Yorkshire boars. Finally, we investigated the efficacy of single/combined markers to predict male fertility using a comprehensive statistical model. Our result showed that there were no significant differences in sperm motility, motion kinematics, or capacitation status, however, the mRNA expression of PSP-I and PSP-II in spermatozoa was significantly different in high- and low-litter size boars. In the individual screening test, the expression of both genes was negatively correlated with boar fertility (r = 0-0.578 and −0.456, respectively), whereas only hyperactivation (HYP) showed a positive correlation (r = 0.444) among the tested semen parameters. As single markers, PSP-I and PSP-II have a better diagnostic power to predict boar fertility, regardless of HYP, in quality assessment analyses. In addition, when these markers were combined, the positive predictive value, negative predictive value, and overall test effectiveness for fertility detection were improved. Surprisingly, when PSP-I and PSP-II were considered together, the deviation of the predicted average litter size between high- and low-litter size boars was 1.77. Based on the findings, we suggest that the use of genomic markers in spermatozoa rather than commonly analyzed semen parameters may be more accurate for evaluating male fertility. Moreover, using a combination of markers could increase the overall accuracy of (in)fertility predictions, and thus, could be considered for field application.

Pengaruh berbagai formulasi pengencer dasar air kelapa terhadap kualitas semen cair sapi PO (Peranakan Ongole) selama simpan dingin

Coconut water extender is the solution to the difficulty and high cots of procurement of diluents. The objective of this study was to examine the effect of various formulations of coconut water extender on quality of PO bull sperm during storage in 4-5 o C. This study was carried out at Animal Reproduction Laboratory of Faculty of Animal Husbandry Brawijaya University and Laboratory of Beef Cattle Research Center Grati Pasuruan on March to April, 2018. The study method used laboratory experimental. The experimental design was Factorial Randomized Completely Block Design and the data were analyzed by Analyze of Variance. There are four treatments in this study (P0 = CEP-3 + 20 % of yolk, P1 = Coconut Water + 20% of yolk, P2 = Coconut Water + 20% of yolk + 0,4% white egg + fructose 1 mg/ml, dan P3 = Coconut Water + 20% of yolk + 0,4% white egg + fructose 2 mg/ml) and ten replications of each treatment. There were significant differences in sperm motility between CEP-3 extender and coconut water-based with various formula extenders during chilled storage at 2-5 o C. But the treatment between coconut water-based formulas did not show a significant difference. Coconut water extenders are able to maintain sperm motility up to day 5 with values above 57%.

Semen characteristics and selected biochemical markers of canine seminal plasma in various seasons of the year

The aim of this study was to evaluate the influence of season on selected qualitative semen characteristics and biochemical markers of canine seminal plasma. Whole ejaculates were collected from 5 crossbred dogs aged 2-8 years. The study covered a period of one year divided into four seasons: spring (March, April, May), summer (June, July, August), autumn (September, October, November) and winter (December, January, February). Semen samples were subjected to macroscopic and microscopic analyses to determine semen volume, total sperm counts and sperm morphology parameters. The study also involved the determination of sperm motility parameters (CASA system), sperm plasma membrane integrity (SPMI, fluorescent staining SYBR-14/PI), sperm mitochondrial membrane potential (MMP, fluorescent staining JC-1/PI) and the ATP content of sperm cells. Total protein content (TPC) and the activity of alkaline phosphatase (AP) and acid phosphatase (AcP) were determined in biochemical analyses of seminal plasma. No significant differences in ejaculate volume, SMPI or ATP content of sperm cells were observed between seasons. The highest total sperm counts were reported in ejaculates acquired in summer and autumn. The lowest MMP values were determined in summer ejaculates. No significant differences in sperm motility (MOT) were observed throughout the experiment, but ejaculates collected in autumn and winter were characterized by the highest progressive motility (PMOT). AP activity and TPC were not significantly affected by season. However, AcP activity levels were significantly lower in autumn than in the remaining seasons. Seasonal variations in the analyzed macroscopic and microscopic parameters of ejaculates and biochemical markers of seminal plasma did not exert a clear negative effect on the quality of canine semen.

Use of cryotubes for the cryopreservation of tambaqui fish semen (Colossoma macropomum)

Tambaqui (Colossoma macropomum) is a freshwater fish of great importance to aquaculture in several South American countries. Recent studies have developed a protocol for semen cryopreservation in 0.25 and 0.5mL straws; however, this technique has limitations for fingerling production at a large scale due to the high fecundity of tambaqui. The main objective of this study was to evaluate the feasibility of using cryotubes (1.6 and 4.5mL) for tambaqui semen cryopreservation. Semen samples were diluted in freezing solution (5% glucose solution, 10% methylglycol, 5% egg yolk), stored in 1.6 and 4.5mL cryotubes, frozen in liquid nitrogen vapor at −175°C and transferred to a cryogenic container at −196°C. The cryotubes were thawed in a water bath at 60°C for 70 or 90s and the motility (total motility – TM; progressive motility – PM; curvilinear velocity – VCL; straight line velocity – VSL and average path velocity – VAP) and the viability of sperm were evaluated. There was no significant difference in sperm motility and viability post-thawing between 1.6 and 4.5mL cryotubes, except for TM (47% and 40%, respectively). Thawing for 90s provided better results, being used in fertilization trials. Although the fertilization rate did not differ between the cryotubes (41–45%), it was significantly lower than that for fresh semen (74%). A strong positive correlation was observed between the sperm motility and fertilization rate (r=0.69–0.89). We conclude that 1.6 and 4.5mL cryotubes have high potential for tambaqui semen cryopreservation when thawed for a minimum time of 90s at 60°C.

English

Twelve indigenous bulls, allocated to two groups comprising six bulls each, were used in the study to determine the effect of dry season nutritional supplementation on scrotal circumference and semen characteristics. One group was supplemented with silage while the other acted as the control. Soundness evaluations were conducted in October, December and February. Breeding efficiency of bulls was measured by pregnancy rates. Month and nutrition significantly affected body condition and body weights in both groups (P &lt; 0.05). Mean scrotal circumference was significantly higher (P &gt; 0.05) for the supplemented bulls than the control group in October (36.3 &plusmn; 1.5 versus 32.0 &plusmn; 2.6 cm). Nutrition and month of collection did not affect scrotal circumference and semen characteristics in both groups (P &gt; 0.05). There were no significant differences in sperm motility between the two groups. Sperm concentration was higher in the control group in all months of collection (P &lt; 0.05). Body condition scores and body weights of cows increased initially but declined to a minimum in December (2.9 &plusmn; 0.34; 298 &plusmn; 15 kg, respectively). Highest mean pregnancy rate (42%) was recorded in Jese village in October while the lowest (13%) was recorded in Makomo village during the same month. It was concluded that supplementary feed did not influence semen characteristics and reproductive capacity of the bulls. &nbsp; Key words: Bull, nutrition, scrotal circumference, semen, breeding soundness evaluation, pregnancy rate, ovarian activity.&nbsp

A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone on the induction of spermiation and amplexus in the American toad (Anaxyrus americanus)

BackgroundCaptive breeding programs for endangered amphibian species often utilize exogenous hormones for species that are difficult to breed. The purpose of our study was to compare the efficacy of two different hormones at various concentrations on sperm production, quantity and quality over time in order to optimize assisted breeding.MethodsMale American toads (Anaxyrus americanus) were divided into three separate treatment groups, with animals in each group rotated through different concentrations of luteinizing hormone releasing hormone analog (LHRH; 0.1, 1.0, 4.0 and 32 micrograms/toad), human chorionic gonadotropin (hCG; 50, 100, 200, and 300 IU), or the control over 24 hours. We evaluated the number of males that respond by producing spermic urine, the sperm concentration, percent motility, and quality of forward progression. We also evaluated the effects of hCG and LHRH on reproductive behavior as assessed by amplexus. Data were analyzed using the Generalized Estimating Equations incorporating repeated measures over time and including the main effects of treatment and time, and the treatment by time interaction.ResultsThe hormone hCG was significantly more effective at stimulating spermiation in male Anaxyrus americanus than LHRH and showed a dose-dependent response in the number of animals producing sperm. At the most effective hCG dose (300 IU), 100% of the male toads produced sperm, compared to only 35% for the best LHRH dose tested (4.0 micrograms). In addition to having a greater number of responders (P < 0.05), the 300 IU hCG treatment group had a much higher average sperm concentration (P < 0.05) than the treatment group receiving 4.0 micrograms LHRH. In contrast, these two treatments did not result in significant differences in sperm motility or quality of forward progressive motility. However, more males went into amplexus when treated with LHRH vs. hCG (90% vs. 75%) by nine hours post-administration.ConclusionThere is a clear dichotomy between the two hormones’ physiological responses on gamete production and stimulation of amplexus. Understanding how these two hormones influence physiology and reproductive behaviors in amphibians will have direct bearing on establishing similar breeding protocols for endangered species.

Boar semen quality of the Přeštice black-pied breed (Short Communication)

Abstract. The objective of this study was to examine semen quality parameters for the Přeštice black-pied breed over a test period of 8 years while considering the potential effects of collection month and boar age. Ninety-nine ejaculates were collected using the gloved-hand technique from healthy and fertile mature boars from selected farms. Ejaculate volumes were relatively low because the boars were accustomed to natural mating. Sperm motility, sperm concentration, percentage of morphologically abnormal spermatozoa (MAS), and sperm motility after 24 h of storage in Androhep extender (dilution rate 1:1.5) were assessed. Significant differences were found in sperm concentration and MAS rate in relation to collection months and boar age in the monitored years (P&lt;0.05). A tendency for MAS to increase with monitored years was observed. Significant differences in sperm motility and motility after 24 h of storage were only observed in relation to collection months (P&lt;0.05). Results of this study detected effects due to collection month and boar age on boar semen quality during the monitored years.