Effect of transfection and co-incubation of bovine sperm with exogenous DNA on sperm quality and functional parameters for its use in sperm-mediated gene transfer

Zygote ◽  
2016 ◽  
Vol 25 (1) ◽  
pp. 85-97 ◽  
Author(s):  
María Elena Arias ◽  
Esther Sánchez-Villalba ◽  
Andrea Delgado ◽  
Ricardo Felmer
Keyword(s):  
Gene Transfer ◽  
Sperm Quality ◽  
Computer Assisted ◽  
Exogenous Dna ◽  
Sperm Analysis ◽  
Functional Parameters ◽  
Transgenic Embryos ◽  
Fertilization Capacity ◽  
Motility Parameters

SummarySperm-mediated gene transfer (SMGT) is based on the capacity of sperm to bind exogenous DNA and transfer it into the oocyte during fertilization. In bovines, the progress of this technology has been slow due to the poor reproducibility and efficiency of the production of transgenic embryos. The aim of the present study was to evaluate the effects of different sperm transfection systems on the quality and functional parameters of sperm. Additionally, the ability of sperm to bind and incorporate exogenous DNA was assessed. These analyses were carried out by flow cytometry and confocal fluorescence microscopy, and motility parameters were also evaluated by computer-assisted sperm analysis (CASA). Transfection was carried out using complexes of plasmid DNA with Lipofectamine, SuperFect and TurboFect for 0.5, 1, 2 or 4 h. The results showed that all of the transfection treatments promoted sperm binding and incorporation of exogenous DNA, similar to sperm incorporation of DNA alone, without affecting the viability. Nevertheless, the treatments and incubation times significantly affected the motility parameters, although no effect on the integrity of DNA or the levels of reactive oxygen species (ROS) was observed. Additionally, we observed that transfection using SuperFect and TurboFect negatively affected the acrosome integrity, and TurboFect affected the mitochondrial membrane potential of sperm. In conclusion, we demonstrated binding and incorporation of exogenous DNA by sperm after transfection and confirmed the capacity of sperm to spontaneously incorporate exogenous DNA. These findings will allow the establishment of the most appropriate method [intracytoplasmic sperm injection (ICSI) orin vitrofertilization (IVF)] of generating transgenic embryos via SMGT based on the fertilization capacity of transfected sperm.

12 COMPUTER-ASSISTED SPERM ANALYSIS OF FRESH EPIDIDYMAL CAT SPERMATOZOA AND THE IMPACT OF COOLED STORAGE (4°C) ON SPERM QUALITY

2008 ◽  
Vol 20 (1) ◽  
pp. 86
Author(s):  
M. Filliers ◽  
T. Rijsselaere ◽  
P. Bossaert ◽  
V. De Causmaecker ◽  
J. Dewulf ◽  
...  
Keyword(s):  
Reference Values ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
The Impact ◽  
Motility Parameters

Feline epididymal sperm is commonly used for in vitro fertilization. It also yields the opportunity to conserve genetic material from valuable males that suddenly die. Epididymal sperm quality parameters vary considerably among laboratories, implicating the need for objective evaluation methods. The aim of the present study was to describe reference values of computer-assisted sperm analysis (CASA) parameters of fresh epididymal cat sperm and to assess the effect of prolonged cooled storage (4�C) on various sample characteristics. Epididymides obtained from tomcats after routine orchiectomy (2–4 pairs/replicate) were sliced to release spermatozoa. The sperm suspension was placed on a 2-layer gradient and, after centrifugation, the sperm pellet was recovered. In Experiment 1 (20 replicates), sperm motility parameters were assessed immediately after retrieval (T0) using the Hamilton Thorne analyzer Ceros 12.1 (HTR; Hamilton Thorne Biosciences, Beverly, MA, USA). In Experiment 2, fresh (T0) sperm samples (4 replicates) were evaluated for motility parameters (HTR), acrosomal status (FITC-Pisum sativum agglutinin staining), morphology (eosin/nigrosin (E/N) staining), and membrane integrity (E/N and SYBR�-14-propidium iodide staining; Molecular Probes, Inc., Eugene, OR, USA). After addition (1:2) of a Tris-glucose-citrate diluent containing 20% egg yolk, samples were cooled and reassessed on Days 1 (T1), 3 (T3), 5 (T5), 7 (T7), and 10 (T10). Results were analyzed in a mixed linear model, with replicate as random factor and time as fixed effect (S-PLUS 7.0; Insightful Corp., Seattle, WA, USA). Results of Experiment 1 were as follows (mean � SD): motility (MOT): 80.8% � 23.5; progressive motility (PMOT): 69.9% � 23.2; velocity average pathway (VAP): 98.7 µm s–1 � 24.2; velocity straight line (VSL): 89.3 µm s–1 � 25.4; velocity curved line (VCL): 134.8 µm s–1 � 31.9; amplitude lateral head (ALH): 4.3 µm � 2.0; beat cross frequency (BCF): 34.6 Hz � 7.0; and straightness (STR): 89.6% � 6.6. In Experiment 2, MOT, PMOT, VAP, VSL, VCL, BCF, and the percentage of normal spermatozoa showed a decrease over time (P < 0.05) compared to fresh samples, starting from T1, T3, T5, T7, T5, T3, and T1, respectively. In contrast, STR, ALH, membrane integrity, and the percentage of acrosome-intact spermatozoa were not affected (P > 0.05) by cooled storage. To summarize, we have presented a set of reference values for CASA-parameters of fresh, epididymal cat spermatozoa. Cooled storage impaired most motility parameters and lowered the percentage of normal spermatozoa, but did not influence membrane integrity or acrosomal status. The effect of cooled storage on DNA fragmentation of sperm and its subsequent influence on in vitro embryo development require further investigation.

scholarly journals Paternal effect on embryo quality in diabetic mice is related to poor sperm quality and associated with decreased glucose transporter expression

Reproduction ◽  
2008 ◽  
Vol 136 (3) ◽  
pp. 313-322 ◽  
Author(s):  
Sung Tae Kim ◽  
Kelle H Moley
Keyword(s):  
Glucose Transporter ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Diabetic Mouse ◽  
Blastocyst Stage ◽  
Computer Assisted ◽  
Diabetic Mice ◽  
Sperm Analysis ◽  
Fertilization Capacity

The objective of this study was to determine whether sperm quality, fertilization capacity, and subsequent embryo development are altered in diabetic male mice and whether differences in facilitative glucose transporter (GLUT; now known as solute carrier family 2, SLC2A) expression in the testis and sperm exist. Using two type 1 diabetic mouse models, SLC2A expression in the testis and sperm was determined by western immunoblotting and immunofluorescence staining. To address sperm quality and fertilization capacity, computer-assisted sperm analysis andin vitrofertilization were performed. SLC2A1, SLC2A3, and SLC2A5 did not change in expression in the testes or sperm between diabetic and non-diabetic mice. SLC2A8 and SLC2A9b were less expressed in the testes of both diabetic models versus controls. SLC2A9a was not expressed in the Akita testis or sperm when compared with strain-matched controls. 3β-hydroxysteroid dehydrogenase (HSD3B) expression was significantly decreased in the Leydig cells from the diabetic mice. Sperm concentration and motility were significantly lower in both the diabetics when compared with the control. These parameters normalized in Akita diabetic males treated with insulin. In addition, fertilization rates were significantly lower in the Akita group (17.9%) and the streptozotocin (STZ)-injected male group (43.6%) when compared with the normal group (88.8%). Interestingly, of the fertilized zygotes, embryo developmental rates to the blastocyst stage were lower in both diabetic models (7.1% Akita and 50.0% STZ) when compared with controls (71.7%). Male diabetes may cause male subfertility by altering steroidogenesis, sperm motility, and SLC2A expression. This is the first study to link a paternal metabolic abnormality to a sperm effect on cell division and subsequent embryonic development.

scholarly journals Effects of chilled storage and pH of activating solution on different motility parameters in burbot (Lota lota) sperm

2018 ◽  
Vol 63 (No. 11) ◽  
pp. 429-434
Author(s):  
Zoltán Bokor ◽  
Balázs Csorbai ◽  
Levente Várkonyi ◽  
Zsolt Szári ◽  
Ferenc Fodor ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Chilled Storage ◽  
Straight Line ◽  
H 26 ◽  
Computer Assisted Sperm Analysis ◽  
Motility Parameters ◽  
Activating Solution

The effects of a simple saline solution prepared using two different pH (4.4 and 8.5) on sperm motility in burbot were investigated. Results were recorded during a 96-hour chilled storage (4°C) in 24-hour intervals. Measurements were focused on the detailed characteristics of motility using 12 parameters obtained from the Computer-assisted Sperm Analysis (CASA). Significantly higher progressive motility (pMOT), distance average path (DAP), distance curved line, distance straight line (DSL), average path velocity (VAP), curvilinear velocity, straight line velocity, and beat cross frequency (BCF) were observed with the activating solution buffered at pH 8.5 in comparison with pH 4.4. Already after 24 h a significant reduction was measured in pMOT (0 h: 49 ± 24%, 24 h: 12 ± 7%). Similar decreasing tendency was recorded only after 72 h in DAP (0 h: 26 ± 4 µm/s, 72 h: 19 ± 9 µm/s), DSL (0 h: 21 ± 5 µm/s, 72 h: 17 ± 8 µm/s), VAP (0 h: 59 ± 9 µm/s, 72 h: 43 ± 21 µm/s), and BCF (0 h: 28 ± 2 Hz, 72 h: 18 ± 10 Hz). The response of different investigated CASA parameters to different treatments varied in our experiments. According to our studies, numerous burbot sperm motility parameters are sensitive to chilled storage and to low pH of the activating solution. Our results could support the effective sperm quality assessment and successful artificial propagation process in burbot.

scholarly journals Melatonin improves rate of monospermic fertilization and early embryo development in a bovine IVF system

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0256701
Author(s):  
Juan Carlos Gutiérrez-Añez ◽  
Heiko Henning ◽  
Andrea Lucas-Hahn ◽  
Ulrich Baulain ◽  
Patrick Aldag ◽  
...  
Keyword(s):  
Embryo Development ◽  
Sperm Quality ◽  
Hierarchical Cluster ◽  
Early Embryo ◽  
Developmental Competence ◽  
Control Group ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Flow Cytometry Data

The developmental competence of male and female gametes is frequently reduced under in vitro conditions, mainly due to oxidative stress during handling. The amino-acid derived hormone melatonin has emerged as a potent non-enzymatic antioxidant in many biological systems. The goal of the present study was to evaluate the effects of melatonin on post-thaw sperm quality, fertilizing ability, and embryo development and competence in vitro after in vitro fertilization. Frozen-thawed bovine spermatozoa were incubated either in the presence of 10−11 M melatonin (MT), or its solvent (ethanol; Sham-Control), or plain Tyrode’s Albumin Lactate Pyruvate medium (TALP, Control). Computer-Assisted Sperm Analysis (CASA) and flow cytometry data after 30 min, 120 min, and 180 min incubation did not reveal any significant effects of melatonin on average motility parameters, sperm subpopulation structure as determined by hierarchical cluster, or on the percentage of viable, acrosome intact sperm, or viable sperm with active mitochondria. Nevertheless, in vitro matured cumulus-oocyte-complexes fertilized with spermatozoa which had been preincubated with 10−11 M melatonin (MT-Sperm) showed higher (P < 0.01) rates of monospermic fertilization, reduced (P < 0.05) polyspermy and enhanced (P < 0.05) embryo development compared to the Control group. Moreover, the relative abundance of MAPK13 in the in vitro-derived blastocysts was greater (P < 0.05) than observed in the Control group. In conclusion, adding melatonin to the sperm-preparation protocol for bovine IVF improved proper fertilization and enhanced embryonic development and competence in vitro.

88 SELECTION OF A CRYOSURVIVAL SPERM POPULATION DOES NOT IMPROVE THE IN VITRO PENETRATING ABILITY OF FROZEN - THAWED BOAR SPERMATOZOA

2008 ◽  
Vol 20 (1) ◽  
pp. 124
Author(s):  
M. L. Perals ◽  
M. A. Gil ◽  
E. M. Garcia ◽  
J. Sanchez-Osorio ◽  
J. M. Vázquez ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Molecular Probes ◽  
Computer Assisted ◽  
Immature Oocytes ◽  
Sperm Analysis ◽  
Boar Spermatozoa ◽  
Casa System ◽  
Selection Of

Boars can be classified as good or bad sperm freezers according to their sperm cryosurvival. Different sperm selection techniques, such as PureSperm� (PS; MidAtlantic Diagnostics, Inc., Mount Laurel, NJ, USA), have been developed to improve functional competence of spermatozoa. The aim of this experimental study was to assess the ability of PS for improving the in vitro penetrating ability of frozen–thawed boar spermatozoa from good and bad sperm freezers. The sperm-rich fractions from two boars, good (Boar A) and bad (Boar B) freezers, were extended in a lactose/eggyolk/ glycerol/Equex Stem (Noba Chemical Sales, Inc., Scituate, ME, USA) mixture (1 � 109 sperm mL–1), dispensed into 0.5-mL straws, and frozen using a programmable cell freezer. After thawing (1.200�C min–1), semen from each boar was split into two aliquots of 500 µL. One aliquot was used as the control. The second was placed into a tube of PS gradient (90%/45%) and centrifuged at 425g for 20 min; the pellet re-suspended in 1 mL of BTS and re-centrifuged at 320g for 10 min (PS sample). Control and PS samples were diluted in supplemented TCM-199 (TCMm; Roca et al. 1998 Reprod. Fertil. Dev. 10, 479–485) at 200 � 106 sperm mL–1. Sperm survival (SV) was assessed afterTCMm dilution according to progressive sperm motility (PSM, %) using a computer-assisted sperm analysis (CASA) system (ISAS�), and plasma and acrosome membrane integrity (PMI; %) by flow cytometry (SYBR�-14/PE-PNA/PI; Molecular Probes, Leiden, The Netherlands). A homologous in vitro penetration (hIVP) assay, using immature oocytes (20 oocytes/2 mL TCMm supplemented with caffeine and calcium lactate), was used to assess sperm penetrating ability (Martinez et al. 1993 Theriogenology 40, 547–557). A total of 960 immature oocytes were inseminated (200 � 103 sperm/oocyte) in 3 batches. After 18 h of co-incubation at 39�C under 5% CO2 in air, the oocytes were washed, mounted on slides, fixed with ethanol:acetic acid (3:1, v/v) for 48 h, stained with 1% lacmoid, and examined under a phase contrast microscope (�400). Oocytes with swollen or unswollen heads of sperm found in the vitellus were considered as penetrated. Sperm penetrability ability (SPA) was assessed according to penetration rate (PR) and the mean number of sperm per oocyte (S/O). Data were analyzed using a PROMIXED model and expressed as mean � SEM. Boar A showed better (P ≤ 0.01) results for both SV and SPA parameters than boar B, independent of sperm treatment. PureSperm improved (P ≤ 0.05) PSM and PMI in both boar A (control v. PS: 48.0 � 5.8 v. 66.5 � 3.6 and 63.1 � 7.7 v. 88.4 � 1.3, respectively) and boar B (12.3 � 1.2 v. 22.2 � 3.7 and 44.3 � 3.5 v. 58.7 � 7.0, respectively). However, no differences (P ≥ 0.05) were observed in PR and S/O in either boar A (71.2 � 3.4 v. 78.3 � 3.1 and 5.0 � 0.4 v. 5.2 � 0.4, respectively) or boar B (34.3 � 3.6 v. 37.3 � 3.9 and 1.5 � 0.1 v. 1.5 � 0.1, respectively). In conclusion, under our laboratory conditions, PureSperm selection improves sperm quality but not in vitro penetrating ability of frozen–thawed spermatozoa of both good and bad sperm freezers. This work was supported by CICYT (AGF2005-00760), Madrid, Spain.

scholarly journals The Effect of Supplementation with Some Essential Oils on the Mobility and the Vitality of Human Sperm

2019 ◽  
Vol 2019 ◽  
pp. 1-6 ◽  
Author(s):  
Modou M. Mbaye ◽  
Bouchra El Khalfi ◽  
Boutaina Addoum ◽  
Papa D. Mar ◽  
Brahim Saadani ◽  
...  
Keyword(s):  
Essential Oils ◽  
Sperm Quality ◽  
Human Sperm ◽  
Optical Microscope ◽  
Computer Assisted ◽  
Vitro Fertilization ◽  
Sperm Analysis ◽  
Improvement Effect ◽  
Sperm Mobility

The objective of this work is to study the improvement effect of some essential oils of sage (Salvia officinalis), oregano (Origanum vulgare), and eucalyptus (eucalyptus globulus) on the physiological parameters characterizing the quality of human sperm (mobility and vitality). We find natural biomolecules to improve sperm quality to increase the chances of success of very low in vitro fertilization (IVF) that stagnate around 20%. Sperm samples were mixed with different concentrations of essential oils. The effect of these essential oils on the motility and vitality of spermatozoa has been analyzed. The mobility was determined using a Computer Assisted Sperm Analysis (CASA). In the other side, the evaluation of sperm vitality was performed by staining eosin 2% and the microscopic examination is carried out via optical microscope. A drop of sperm will be mixed with a drop of eosin solution 2%, spread between the slip and coverslip, then allowed to air dry, and examined under a microscope. A significant improvement in the mobility and vitality of human spermatozoa has been noted with oregano. Eucalyptus after 10 min of exposure also significantly improves the mobility and vitality of the spermatozoa. Sage does not improve mobility for these incubation times but significantly improves vitality.

scholarly journals 238 ASSESSMENT OF SPERM MOTILITY, VIABILITY, AND FERTILIZATION RATE OF TWO GROUPS OF BULLS SHOWING A DIFFERENT ABILITY TO PROMOTE EMBRYO DEVELOPMENT IN VITRO

2005 ◽  
Vol 17 (2) ◽  
pp. 269 ◽  
Author(s):  
M. Alomar ◽  
J. Mahieu ◽  
B. Verhaeghe ◽  
I. Donnay
Keyword(s):  
Low Fertility ◽  
Computer Assisted ◽  
High Fertility ◽  
In Vitro Production ◽  
Sperm Analysis ◽  
Fertilization Rates ◽  
Significant Difference ◽  
Sperm Nuclei ◽  
Motility Parameters

Low developmental rates after IVF with some AI bulls are one of the most frequent problems encountered with in vitro production of bovine embryos. In this study, two groups of high and low in vitro fertility bulls (3 bulls per group) were selected based upon blastocyst rates. We first evaluated sperm motion characteristics using computer-assisted sperm analysis (CASA) before and after Percoll separation. The following parameters were compared between bulls: proportion of motile sperm (% MOT), curvilinear velocity (VCL), and straightness (STR), which allowed us to evaluate the proportion of progressive semen (VCL > 80 μm/s and STR > 60%). Concerning % MOT, differences between bulls were observed before Percoll separation (42 to 59% – ANOVA2: P = 0.017). Following separation, the values of % MOT were globally increased (ANOVA3: P < 0.001) and no further difference was observed between bulls (68 to 76%). The same pattern was obtained for the proportion of progressive spermatozoa (before: 43 to 55% vs. after: 48 to 57% – ANOVA3: P = 0.005). For viability measurements, two DNA-binding fluorochromes, propidium iodide and Hoechst 33342, were used, which respectively stain dead sperm nuclei in red and living sperm nuclei in blue. This viability evaluation was carried out after 0, 2, 6, and 18 h of incubation in TALP medium supplemented with heparin (10 μg/mL), under the same conditions as IVF but without oocytes. In the two groups of bulls, a sharp decrease in the proportion of living sperm was already observed at 2 h (only 50 to 20% of living sperm), while, at 18 h, 17 to 9% of sperm were still intact. Two bulls from the high in vitro fertility group showed a percentage of living sperm significantly more important at 6 and 18 h than the other 4 bulls (chi square – P < 0.05). In a third experiment, we assessed the fertilization (2 pronuclei) and the polyspermy (>2 pronuclei) rates after staining the zygotes with Hoechst at 18 hpi. The high fertility group showed higher fertilization rates than the low fertility one (75 to 80% vs. 59 to 67% – ANOVA2: P < 0.008). No significant difference was noticed between bulls concerning polyspermy. The present results confirm the efficiency of Percoll separation in enhancing some of the motility parameters such as the proportion of motile and progressive sperm. Moreover, this gradient allows us to abolish differences between bulls for those motility parameters. However, the viability and the rate of fertilization still varied between bulls after Percoll separation. In our study, only fertilization rates seemed related to the blastocyst rates. Further investigations are under way to better understand the sperm characteristics of these two groups. This work was supported by the Minestery of Agriculture of the Region wallonne de Belgique.

scholarly journals Effects of Various Incubation Conditions on Functional Parameters of Stallion Spermatozoa

2018 ◽  
Vol 49 (3) ◽  
pp. 201-208 ◽  
Author(s):  
O. Šimoník ◽  
J. Šichtař
Keyword(s):  
Cluster Analysis ◽  
Incubation Time ◽  
Computer Assisted ◽  
Free Atmosphere ◽  
Sperm Analysis ◽  
Functional Parameters ◽  
Mitochondrial Integrity ◽  
Computer Assisted Sperm Analysis ◽  
Sample Dilution

Abstract The objective of our study was to determine the effect of 5% of CO2 atmosphere and time of sample dilution on results of in vitro analysis of stallion semen. Frozen-thawed semen from 14 stallions was incubated either in incubator or in a water bath, diluted prior to analysis or immediatelly after thawing. The following qualitative parameters were assessed after thawing (T0) and after 30 min (T30): motility in 3 sperm subpopulations (slow, medium, fast) defined by cluster analysis of parameters obtained by Computer Assisted Sperm Analysis, viability, acrosome and mitochondrial integrity. The slow subpopulation was only significantly reduced in diluted samples in CO2 atmosphere at T0 (P < 0.05). In diluted samples the incubation time significantly affected distribution of fast, slow, and medium subpopulations in CO2 and CO2 free atmosphere (P < 0.05), respectively. Viability, acrosome and mitochondrial integrity were not affected by CO2 atmosphere (P > 0.05), however acrosome (at T0) and mitochondrial integrity (at T30) were significantly higher in CO2 atmosphere in non-diluted and diluted samples (P < 0.05), respectively. The results of the in vitro analysis of stallion semen were rather similar regardless of the atmosphere or dilution time.

scholarly journals Post-Thaw Sperm Quality and Functionality in the Autochthonous Pig Breed Gochu Asturcelta

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1885
Author(s):  
José Néstor Caamaño ◽  
Carolina Tamargo ◽  
Inmaculada Parrilla ◽  
Felipe Martínez-Pastor ◽  
Lorena Padilla ◽  
...  
Keyword(s):  
Genetic Resource ◽  
Sperm Quality ◽  
Genetic Material ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Linear Mixed Effects ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Analysis System

Genetic resource banks (GRB) preserve the genetic material of endangered, valuable individuals or genetically relevant breeds. Semen cryopreservation is a crucial technique to reach these goals. Thus, we aimed to assess the sperm parameters of semen doses from the native pig breed Gochu Asturcelta stored at the GRB of Principado de Asturias (GRB-PA, Gijón, Spain), focusing on intrinsic and extrinsic (boar, season) factors. Two straws per boar (n = 18, 8–71 months of age) were thawed, pooled, and assessed after 30 and 150 min at 37 °C by CASA (computer-assisted sperm analysis system; motility and kinematic parameters) and flow cytometry (viability, acrosomal status, mitochondrial activity, apoptosis, reactive oxygen species, and chromatin status). The effects of age, incubation, and season on post-thawing quality were determined using linear mixed-effects models. Parameters were on the range for commercial boar breeds, with chromatin status (SCSA: fragmentation and immaturity) being excellent. Incubation decreased sperm quality and functionality. The boar age did not have a significant effect (p > 0.05), but the between-boar variability was significant (p < 0.001). The season significantly affected many parameters (motility, kinematics, viability, acrosomal status, mitochondrial activity), especially after 150 min of incubation. In general, samples collected in spring and summer showed higher quality post-thawing, the lowest in winter. In conclusion, the sperm doses from the Gochu Asturcelta breed stored at the GRB-PA showed excellent chromatin status and acceptable characteristics after thawing. Therefore, boar and seasonal variability in this autochthonous breed could be relevant for cryobank management.

scholarly journals Effect of taurine on turkey (Meleagris gallopavo) spermatozoa viability and motility

2018 ◽  
Vol 63 (No. 4) ◽  
pp. 127-135 ◽  
Author(s):  
T. Slanina ◽  
M. Miškeje ◽  
F. Tirpák ◽  
M. Błaszczyk ◽  
R. Stawarz ◽  
...  
Keyword(s):  
Meleagris Gallopavo ◽  
Lower Percentage ◽  
Computer Assisted ◽  
Viability Assessment ◽  
Spermatozoa Motility ◽  
In Vitro Incubation ◽  
Time Periods ◽  
Casa System ◽  
Motility Parameters

The effect of taurine on the turkey spermatozoa motility and viability during the in vitro incubation was assessed. Experimental samples were prepared by diluting the raw semen in nine different concentrations of taurine – from 10 mg/ml to 0.078125 mg/ml. The motility parameters were evaluated by the CASA system (Computer Assisted Semen Analyser) using the program Sperm Vision<sup>®</sup> and for spermatozoa viability assessment the eosin-nigrosin staining was performed. Selected parameters were evaluated at six time periods: 0, 1, 2, 3, 4, and 5 h at 5°C and 41°C. At 5°C, a significantly lower percentage of motility and progressive motility was detected only in the samples with the highest concentration of taurine (10 mg/ml) at time 0 and 1. After 2 h of incubation a significant preventive effect of taurine on spermatozoa parameters was observed. The tendency of the taurine effect on motility parameters was different during the in vitro incubation at 41°C. Significantly lower values of motility parameters were detected in all experimental samples in comparison to the control after 5 h. The analysed concentrations of taurine did not significantly affect viability of turkey spermatozoa during all time periods. A higher percentage of dead spermatozoa were observed at 41°C (4.87–9.90%) if compared to 5°C (2.12–4.88%). The results indicated that the addition of taurine (from 2.5 to 7.5 mg/ml) to turkey spermatozoa positively affected the monitored spermatozoa parameters incubated at 5°C.

Sign in / Sign up

Export Citation Format

Share Document