Higher Percentage Of Motility Research Articles

Do milt storage techniques affect the quality of carp spermatozoa during short-term storage?

Short-term storage and management of milt are crucial parts of hatchery practice. There is speculation that sperm stored in a small Eppendorf could not retain their optimal performance compared to those stored in a 50 ml cell culture flask (CCF), thus raising the question of discrepancy between laboratory and hatchery conditions. Therefore, this study aimed to compare sperm quality parameters, such as osmolality and the ionic composition of seminal plasma, spermatozoa motility and its ability to maintain the volume under various osmotic conditions (310, 230, 150, 90, and 20 mOsm/kg) in fresh sperm and after its storage for 24 h in 1.5 ml Eppendorf or 50 ml CCF at 4 °C without any extender. Freshly collected sperm had higher motility percentages at each osmotic condition than 24 h stored sperm in Eppendorf and CCF. There was no significant difference in spermatozoa motility at various osmotic conditions except 90 mOsm/kg osmolality. Besides, VCL, VSL, VAP, and LIN of spermatozoa, osmolality, and ionic composition of seminal plasma did not significantly differ between these two containers. This study suggested no difference between sperm stored in an Eppendorf or CCF regarding sperm functionality.

The Effect of Additional Tomato Extract in Diluting Sodium Citrate and Egg Yellow in Kampung Chicken on The Quality of Semen Cow Limousine

Undiluted sperm will reduce fertility, therefore dilution is performed to maintain the quality of spermatozoa during storage. Tomatoes contain carbohydrates, vitamin A, vitamin C, protein, and lycopene as antioxidants. Carbohydrates and antioxidants act as a source of energy to potentially prevent free radicals that can damage cells. This study aims to determine the effect of adding tomato juice in sodium citrate diluent and free-range chicken egg yolks to the semen quality of limousin cattle. This study used a completely randomized design (CRD) with 4 treatments and 5 replications. The treatment consisted of a solution of sodium citrate and egg yolk (P0), sodium citrate and egg yolk + 10% tomato extract (P1), sodium citrate and egg yolk + 20% tomato extract (P2), and egg yolk sodium citrate + 30% tomato extract. % (P3). Parameters observed were motility, viability and abnormalities of semen spermatozoa of Limousin cattle. Observations were made at 0, 24, 48, and 72 hours after cooling. The results showed that the addition of 10% tomato extract (P1) had a significant effect on the highest percentage of motility, namely 42.00 ± 2.73c for the 48th hour, the highest viability 62.10 ± 4.51c for the 72nd hour, and lowest abnormality 16.60 ± 1.81a until the 72nd hour. The addition of 10% tomato extract (P1) in sodium citrate diluent and egg yolk resulted in the best motility, viability, and abnormality percentage of Limousin cattle spermatozoa.

Some recent data on sperm morphology and motility kinetics in Atlantic cod (Gadus morhua L.).

Studying biology of sperm provides valuable information to optimize artificial reproduction and is crucial for sustainable aquaculture. Here, we investigated morphology of spermatozoon in Atlantic cod (Gadus morhua) using transmission and scanning electron microscopy. Furthermore, spermatozoa motility kinetics at different osmolalities were studied using computer-assisted sperm analysis software. The spermatozoon lacked an acrosome and consisted of a head, midpiece, and flagellum. The head of spermatozoa was round, oval, and rather elongated in shape, showing high variations in dimensions. There were up to 6 mitochondria that encircled the proximal part of the flagellum. The proximal and distal centrioles were located within the nuclear notch and arranged orthogonal to each other. The axoneme had a typical 9 + 2 microtubule structure. The flagellar length of spermatozoon was 66.94 ± 0.46μm. Spermatozoa were immotile in the seminal plasma. Dilution of sperm with natural seawater (1100mOsmol/kg) resulted in initiation of motility for 91.0 ± 3.4% of spermatozoa with average velocity of 86.2 ±2.3μm/s and beating frequency of 52Hz. The duration of spermatozoa motility was > 6min; however, the percentage of motile spermatozoa decreased at 60s post-activation. When osmolality of natural seawater was modified using distilled water or NaCl, spermatozoa motility was not initiated at ≤ 400 and ≥ 2500mOsmol/kg, and the highest percentage of motility was observed at 730-1580mOsmol/kg. In a sucrose solution, spermatozoa motility was initiated and suppressed at 600 and 1500mOsmol/kg, respectively, and highest percentage of motility was observed at 800-1100mOsmol/kg. Spermatozoon morphology comparisons within Gadiformes showed differences in dimensions of head and mitochondria, flagellar length, and number of mitochondria. The present study provides valuable data that can be used for phylogenetic implications based on spermatozoon morphology and for development of artificial fertilization and sperm cryopreservation protocols based on sperm motility.

The Effect of Glutathione on The Quality of Aceh Local Catfish (Clarias gariepinus) Spermatozoa After Cryopreservation

The cryopreservation process might reduce the quality of spermatozoa due to an increase in the production of reactive oxygen species (ROS) compounds during cooling, freezing, and thawing. The quality of spermatozoa can be maintained by adding glutathione as an exogenous antioxidant into cryo-diluent. This study aimed to examine the effect of the addition of different doses of glutathione in cryopreservation of Aceh Local catfish (Clarias gariepinus) spermatozoa after freezing. The method used was a completely randomized design (CRD) with four treatments and four replications. Fresh semen was diluted in Ringer, 15% DMSO, and 20% Fetal Bovine Serum (FBS) and then added with glutathione antioxidants of 0.0 mgL-1, 0.5 mgL-1, 1.0 mgL-1, and 2.0 mgL-1. The parameters observed in this study were motility, integrity of the plasma membrane, fertility, and DNA integrity. The results showed that the concentration of glutathione had no effect on motility, integrity ofthe plasma membrane, or fertility, but had an effect on DNA integrity. The highest percentage of motility and plasma membrane integrity respectively was 40.50% (P3) and 70.87% (P2). Furthermore, the assessment of DNA integrity showed that there was no DNA fragmentation both treatments and fresh spermatozoa. This research is the first study regarding glutathione supplementation in cryo-diluent of Aceh Local catfish spermatozoa. Finally, the results obtained can provide information about the exact concentration of glutathione in the extender on the quality of spermatozoa of Aceh Local catfish (C. gariepinus) after the cryopreservation process. These results can also increase the success of fertility be used by the seed hall unit and the aquaculture industry to increase the productivity and supply high quality seeds.

PENGARUH PENAMBAHAN EKSTRAK TEH HIJAU (Camellia sinensis) DALAM BAHAN PENGENCER SUSU SKIM KUNING TELUR TERHADAP KUALITAS SPERMATOZOA DOMBA SAPUDI YANG DISIMPAN PADA SUHU DINGIN

The purpose of this study was to determinated the effect and the best concentration of greentea extract in skim milk and egg yolk diluent for quality sapudi sheep spermatozoa were measured in motility, viability and intact plasma membrane that was stored on cold temperature. The semen was devided into four groups; skim milk and egg yolk diluent, 0,5% green tea extract in skim milk and egg yolk diluent, 0,1% green tea extract in skim milk and egg yolk diluent, and 0,15% green tea extract in skim milk and egg yolk diluent. Spermatozoa quality was observed day 1, day 2, day 3, day 4 and day 5 after being diluted. The data obtained was analyzed with the Analysis of Variant (ANOVA), followed by multiple range test duncan. The results showed that the highest percentage of motility, viability and intact plasma membrane derived from green tea extract 0,15% on skim milk and egg yolk diluent for 1 day storage is 86.66b ± 2.58, 92.00b ± 1.78, and 74.16b ± 3.25. The lowest percentage of motility, viability and intact plasma membranes was obtained from the skim milk and egg yolk diluent for 5 days storage is 43.33a ± 6.05, 56.50a ± 3.08 and 28.33a ± 2.80. The conclusion of this study is the addition of 0.15% green tea extract in a skim milk and egg yolk diluent can maintain the quality of sperm for up to 5 days of storage at cold temperatures.

Kısa Süreli Saklanan Koç Spermasına İlave Edilen Sığır Serum Albumininin (BSA) Farklı Dozlarının Sperma Kalitesi ve DNA Hasarı Üzerine Etkisi

In the present research the aim was emphasized to find out the effects of bovine serum albumin (BSA) on quality of sperm and DNA damage at 0, 24 th, 48 th and 72 th hours in Merino ram semen at short term preservation (5°C). Ejaculates were obtained from five Merino Rams which were collected by artificial vagina and all the samples were pooled at 37°C for detection. All the procedures were applied six times concurrently. Different concentrations (0, 2, 4 and 6 mg/ml) of BSA with Tris-based extender was used for dilution on semen samples. Sperm motility, abnormality, plasma membrane integrity, acrosome integrity and % DNA in tail (COMET assay) content were measured and analyzed. The extender with 4 mg/ml of BSA had both higher motility percentages and plasma membrane integrity, when compare with the control, up to 72 hours (P<0.05). At the 24th and 48th hours of the study, DNA damage was lower in all doses of BSA than in the control group. In addition, in terms of abnormal spermatozoon rates, all doses of BSA were lower than the control group at 24, 48 and 72 hours (P<0.05). In conclusion, it was found to be especially useful to add 4 mg / ml BSA for the ram semen parameters in short term storage.

Comparison of sperm selection techniques in donkeys: motile subpopulations from a practical point of view.

The aim of this study was to compare the post-thaw distribution of motile sperm subpopulations, following simple or colloid centrifugation. A new analysis was used to evaluate the available number of sperm from each subpopulation after each centrifugation protocol. Frozen/thawed semen samples were divided into the following after-thawing treatments: uncentrifuged control (UDC), sperm washing (SW) and two colloid centrifugation procedures (Equipure, SLC-E, and Androcoll, SLC-A). Percentage of total and progressive motility (TM and PM), as well as sperm motility kinematics, distribution of motile sperm subpopulations, and recovery rates, were statistically compared among treatments. The SLC treatments showed higher (P < 0.001) TM and PM than UDC and SW. Following each SLC procedure, different percentages of the subpopulation with the most vigorous and progressive sperm (sP4) were obtained. SLC-A recovered a larger number of sperm belonging to sP4 than SLC-E, but not significantly higher than SW. From a practical point of view, sperm washing, the standard centrifugation procedure for equine semen processing, recovered the same amount of fast and progressive sperm as colloid centrifugation, apparently the best treatment according to traditional analysis. In conclusion, samples processed by SLC have higher motility percentages than SW and UDC but, after combining the available number of sperm, SLC and SW techniques are equally efficient in recovering sperm from the most vigorous, fast and progressive motile subpopulation (sP4).

34 The effect of dilution method of beagle dog semen on the survival rate of cryopreserved spermatozoa after thawing

The successful cryopreservation of spermatozoa of the beagle dog for AI is essential for the establishment of the genetic banks of drug detection dogs. The beagle dog is widely used for drug testing and chosen for breeding by breeders. However, the use of cryopreserved beagle semen is limited by the lower number of offspring of dog species. In this study, 3 highly trained beagle dogs were chosen and their semen was cryopreserved for the next generation. The effects of dilution methods of beagle semen were tested using a direct dilution method at RT and a 2-step dilution method at 5°C. As a control group, the effects of a direct dilution method of semen on the percentage of motile sperm and progressive motility were analysed by computer-assisted semen analysis system (SAIS, Korea), and abnormality of spermatozoa was examined by Diff Quik staining. A total of 9 samples from 3 dogs were extended in 4% glycerol containing Tris-egg yolk diluents at approximately 22 to 25°C. The diluted semen was cooled to 5°C within 2h. The packed 0.5-mL straws were placed 5cm above the surface of LN for 10min and then plunged in. A 2-step dilution method was conducted using the same procedures of freezing, but the first dilution was done with glycerol-free diluent. After cooling to 5°C within 2h, the second diluent with 8% glycerol was added to the same volume of diluted semen at 5°C and stabilised for 1h. After thawing for 45s at 37°C, the semen from the 2-step dilution method showed the higher percentage of motile sperm (65.4±6% v. 45.3±8%; P&amp;lt;0.05) and progressive motility (41.6±5.3% v. 32.3±3.7%; P&amp;lt;0.05). However, the abnormalities between groups showed no differences. The results suggest that the optimal method for freezing beagle dog spermatozoa is a 2-step dilution process that consists of the first dilution at RT and the second dilution with glycerol at 5°C into diluted semen.

PENGARUH WAKTU INKUBASI PADA PROSES SEXING SPERMA BERBASIS GLUTATHIONE TERHADAP MOTILITAS DAN MEMBRAN PLASMA UTUH CHILLED SEMEN DOMBA LOKAL

The aim of this study was to examine the effect of incubation time on sexing process based on glutathione to the motility and Intact Plasma Membrane (IPM) of chilled ram semen. The separation of X and Y sperm conducted by using BSA method. This research was experimental research using completely randomized design (CRD) with three treatment namely, 45 minutes (T1), 60 minutes (T2) and 75 minutes (T3). Data were analyzed using Anova followed by Duncan's multiple range test. Material used in this research was the fresh semen from ram with sperm motility ≥ 70% and 5 mM of glutathione. The result of this research showed that the highest percentage of motility in upper and bottom fraction belong to T1 (69,7% dan 68,8%), followed by T2 (66,4% dan 64,5%) and T3 (57,9% dan 57,6%). In addition, the result of this research that the highest percentage of IPM in upper and bottom fraction belong to T1(75% dan 71,3%), followed by T2 (69,9% dan 68,4%) and T3 (66,9% dan 65,5%). It can be concluded that the incubation time of 45 minute is the optimum time on sexing process based on glutathione so that the motility and IPM of chilled ram semen can be maintained.

Sperm characteristics of wild and captive lebranche mullet Mugil liza (Valenciennes, 1836), subjected to sperm activation in different pH and salinity conditions

In this article we describe basic aspects of the sperm biology of lebranche mullet (Mugil liza) in the wild and in captivity, in particular assessing the effects of salinity (0, 10, 20, 30, 35, 40, 50 and 60 g L−1) and pH (6, 7, 8, 9 and 10) on sperm motility. Our results indicate that the highest percentage of motility was recorded with salinity 34.6 g L−1 (95 ± 10%) and the longest motility time was obtained with a salinity of 34.8 g L−1 (189 ± 15 s). Variations in the salinity between 30 and 35 g L−1 did not produce any significant alterations in sperm motility; however salinities of 20 and 50 g L−1 produced a significant loss of sperm motility. The highest percentage of motility was obtained at pH 8.5 (93 ± 12%), and the longest motility period at pH 8.7 (218 ± 13 s), while pH lower than or equal to 7 and equal to 10 both produced a significant loss in sperm motility. A positive correlation was found between pH/salinity and the motility percentage (R2 = 0.94 and R2 = 0.97) and motility time (R2 = 0.86 and R2 = 0.98). In seminal and morphometric parameters, statistically significant differences were observed in semen volume, sperm density, plasma membrane integrity and sperm morphometry between the groups studied, showing that the characteristics of the fish have a direct influence on sperm quality. The information generated in this research will be useful for developing biotechnology tools for the effective management of Mugil liza gametes.

Influence of Different Pulse Length towards Motility and Viability of Ornamental Japanese Carp (Cyprinus carpio Var. Koi) Sperm through Electroporation Method

Successful studies about implementation of electroporation method which sperm becomes gene transfer medium for fish transgenesis have reported. Motility and viability are two major factors in successful of electroporation. The objective of the study was to determine optimal pulse length of electroporation towards the motility and viability of Koi fish sperm. The study was conducted at the Central Laboratory of Biological Sciences and Reproduction of Brawijaya University. The data analysis was conducted using completely randomized design. The electroporation method was carried out using voltage of 30 volt with pulse number of 4 times. The pulse length of the electroporation (based on treatment) was 0.5, 1 and 1.5 ms. The result showed the highest percentage of motility was 3.67% in treatment A (pulse length 0.5 ms) with coefficient correlation R 2 = 0.9643 and the best percentage of viability was 79.67% in treatment A (pulse length 0.5 ms) with coefficient correlation R 2 = 1. Correlation between the pulse length treatment (electroporation) toward motility and viability of Koi fish sperm, in which longer duration of pulse length treatment (electroporation) would decrease the motility and viability percentage of the fish sperm. Keywords: Electroporation, Motility, Pulse length, Sperm, Viability.

Thawed human sperm quality is influenced by the volume of the cryopreserved specimen

To test the effect of sperm specimen volume in the freezing-thawing process on specimen quality. Experimental prospective study. Tertiary academic medical center. Fifty high-quality sperm donors donated ∼3 times each. Sperm samples were split into two aliquots and frozen in volumes of 0.25 mL and 0.5 mL. Semen analyses. Eight sperm quality parameters of thawed specimens. Thawed 0.5-mL specimens had a higher percentage of motility and viability, progressive motility concentration, percentage of cells with high mitochondrial membrane potential, and intact chromatin compared with 0.25-mL specimens. Although there were fewer cells with intact acrosomes in the 0.5-mL thawed samples, they had a similar ability to respond to ionophore by acrosome reaction as the 0.25-mL specimens. Both groups had similar percentages of cells with oxidative stress and numbers of cells that bound to the zona pellucida. The remaining air volume in the straw and freezing medium composition had a minimal effect on tested parameters. Better quality thawed human sperm was achieved after cryopreservation of high volumes compared with low volumes of specimens. Air volume in the straw had no influence on specimen quality.

Methionine supplementation improves ram sperm parameters during liquid storage at 5 °C

The aim of this study was to investigate the effects of methionine and lipoic acid on ram sperm parameters during liquid storage (5°C). Ejaculates collected from five Merino rams were pooled at 37°C. Each pooled ejaculate was divided into five equal aliquots and diluted (37°C) with five extenders, one of which was without additives, two of which contained methionine at two different doses, and the other two of which contained lipoic acid at two different doses. Sperm parameters were determined at 0, 24, 48, 72 and 96h of liquid storage at 5°C.The extenders containing 2 and 4mM of methionine resulted in higher motility percentages, in comparison to the control, up to 96h of storage. Methionine at doses of 2 and 4mM led to higher viability and sperm mitochondrial activity percentages, when compared to the controls during 48, 72 and 96h of liquid storage (P<0.05). The findings of this study showed that methionine was of greater benefit to ram sperm parameters during liquid storage.

Abstract 2424: Conditioned media enhances proliferation and mobility in triple negative breast cancer cells

Abstract Background. Triple negative breast cancer (TNBC) is one of the most aggressive forms of breast cancer and disproportionally affects African American women. This subtype of breast cancer is characterized by the absence of the estrogen, progesterone, and HER2/neu receptors. Our research has provided insight on understanding that the microenvironment, specifically noncancerous cells, play in enhancing cancer progression and metastasis. However, the role of adjacent noncancerous cells in the metastatic phenomenon is poorly understood. Therefore, the aims of this project were: 1) to determine if conditioned media from MCF-10A cells would enhance proliferation of MDA-MB-231 and MDA-MB-468 cells and 2) to evaluate the ability of the effects of migration of the conditioned media on the MDA-MB-231 and MDA-MB-468 cells. Methods. Crystal violet and alamar blue dye exclusion assays were performed to measure proliferation in the different medias. The trans well migration assay was used to evaluate motility enhancement based on the media used, either conditioned media and the Cellometer was used to evaluate whether the MDA-MB-468 and MCF-10A cells shared the same microenvironment. Results. Conditioned media is made from base medium with a concentration of 1.0 g/L glucose with .5% FBS exposed to non-cancerous MCF-10A cells that have been driven into crisis. This conditioned media encouraged robust and continuous proliferation of the tumorigenic MDA-MB-231 and MDA-MB-468 cell lines. HGCM was extracted in our lab using sterile technique and reused to analyze the proliferative effects on cancerous and non-cancerous cells. MDA-MB-231 cells demonstrated significant proliferation when compared to the control. MDA-MB-468 cells demonstrated a similar response. The trans well migration assay demonstrated that the GFP labeled MDA-MB-468 cells had a higher percentage of motility than with MCF-10A cells in standard media. Conclusions: Taken together, these data indicate that factors secreted into the microenvironment from noncancerous cells may contribute to the mobility of TNBC cells and the conditioned media contains unknown factors that promote the cell mobility of the MDA-MB-231 and MDA-MB-468 cells. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2424. doi:1538-7445.AM2012-2424

The effects of commercial preparations containing two different GnRH analogues and dopamine antagonists on spermiation and sperm characteristics in the European smelt Osmerus eperlanus (L.)

Efficiency of Ovopel (mGnRHa + metaclopramide) and Ovaprim (sGnRHa + domperidone) treatment on the quantity and quality of European smelt milt were tested. Either preparation resulted in stimulation of smelt sperm production (100% spermiating males vs. 68% in control). At the dosage tested, Ovaprim treatment increased the volume of milt and total sperm production, whereas Ovopel treatment produced sperm with a higher percentage of motility. Among computer-assisted sperm motility analysis (CASA) parameters only amplitude of lateral head displacement (ALH) significantly differed from control in both hormonal treated groups. For most parameters no significant differences between control and stimulated males were found. Our data suggested that administration of two different GnRHa combined with two different dopamine antagonists would help in the future work for improving procedures of artificial reproduction and conservation of European smelt.

In vitro Production of Nitrite by Low and High Density Sperm Subpopulations of Human, Bull and Ram

The aim of present study was to characterize nitric oxide production via nitrite measurement as its stable metabolite in the different species of spermatozoa (human, bull and ram). Sperm samples were fractionated on the basis of density with the use of a three-layer Percoll gradient. Spermatozoa collected from the various percoll gradient interfaces (50/70 and 70/95%). Results showed that nitrite is produced by human, bull and ram spermatozoa, but in the same sample, there were no significant difference between high density (with higher percentage of motility) sperm subpopulation and low density (with lower percentage of motility) sperm subpopulation Therefore, it seems that spermatozoa with different motility in the one sperm population have the same production of NO in physiological conditionthat is different from pathological condition (e.g., in the subfertile men).

THE INFLUENCE OF DIFFERENT CENTRIFUGATION REGIMES ON DOG SPERMATOZOA

The present study has the aim of evaluating the cen trifugation effects over canine semen. The main objectives are establishment of the four centr ifugation regime (650xg/2min., 650xg/5min., 1500xg/2min., 1500xg/5 min.) effects upon spermatozoa by establis hing of quantitative and qualitative losses of sper matozoa in centrifugation stage. Centrifugation regime 1500xg for 5 minutes showed the lowest losses of spermatoz oa: 4,42%. A short semen centrifugation time (2 min) is correlated to a higher motility percentage. Centri fugation regime 650xg for 2 minutes is the most suitable und er the aspect of spermatozoa morphological integrit y maintaining. The functional integrity of the sperma tozoa membranes is less affected by the short centr ifugation time .

Electron paramagnetic resonance spectroscopy for the investigation of the fluidity of human spermatozoa plasma membranes: a feasibility study

Spermatozoal membrane perturbations may play a role in abnormal sperm functions. The objective of this investigation was to study the feasibility of measuring membrane fluidity of isolated human sperm by electron paramagnetic resonance (EPR) spectroscopy and to compare the order parameter of spectra obtained from the sperm plasma membranes of living sperm of fertile men with that of infertile men. Ejaculates of infertile and fertile men were washed and the spermatozoa labelled with 5-doxylstearic acid (5-DSA) and 16-doxylstearic-acid (16-DSA) (10 nmol per 4 x 10(7) sperm). The reporter group of 5-DSA partitions into the outer, hydrophilic part of the sperm plasma membrane, whereas that of 16-DSA is distributed in the inner hydrophobic part. The following results were obtained: (i) the lowest measurable cell count was 3.6 to 7 x 10(6) sperm and the interassay variance of the order-parameter s was < 1%; (ii) swim-up experiments revealed a higher fluidity of sperm with a higher percentage of motility; (iii) sperm membranes of infertile patients exhibited a decreased fluidity of their plasma membranes in the polar interface region of 5-DSA compared with volunteer semen donors and fertile men (P=0.002). No difference of membrane fluidity was found between the different groups using 16-DSA. It is concluded that EPR spectroscopy can be used to study the fluidity of sperm plasma membranes in fertile and infertile men.