118 Semen quality and fertilization ability of myostatin-knockout boars

2020 ◽  
Vol 32 (2) ◽  
pp. 186
Author(s):  
M.-F. Xuan ◽  
S.-Z. Han ◽  
B.-H. Quan ◽  
X.-J. Yin ◽  
J.-D. Kang
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Mitochondrial Activity ◽  
Human Consumption ◽  
Computer Assisted ◽  
Plasma Membrane Integrity ◽  
Sperm Plasma Membrane ◽  
Student’S T

Myostatin-knockout (MSTN−/−) pigs may provide a source of healthy lean protein for human consumption. However, little is known about the effect of this knockout on semen quality, which is important if these pigs are used for production. The purpose of this study is to evaluate the semen quality and fertility of MSTN−/− boars. We generated MSTN−/− boars from Duroc-Landrace-Yorkshire hybrid pig cell lines by somatic cell nuclear transfer, and all 12 boars showed sexual maturation with an obvious “double muscling” phenotype. Semen was collected randomly from three MSTN−/− boars using the gloved-hand technique by one technician and then tested by computer-assisted semen analysis. Semen acrosomal integrity and deformity were measured using Coomassie blue- and eosin-stained smears, respectively. Sperm plasma membrane integrity and mitochondrial activity were evaluated by Hoechst 33342, propidium iodide, and JC-1 multiple staining. The reproductive performance of MSTN−/− boars was evaluated by IVF and by AI. All data were analysed by Student's t-tests. The results showed that the semen color, odor, and pH had no abnormalities. The concentration, motility, plasma membrane integrity, deformity, acrosome integrity, and mitochondrial activity of the semen presented no significant differences from those of the control semen (Duroc). The ejaculation volume of the MSTN−/− boars was significantly lower than that of the control (168.78±6.70 and 223.11±21.21mL, respectively), although the total sperm number was not significantly different. The rate of cleavage and blastocyst formation (247 to 254 oocytes per boar) was not significantly different from those of the control (69.1±0.7 vs. 65.2±1.6%, and 20.2±1.2 vs. 22.8±1.4%, respectively). Seventeen healthy offspring were successfully produced from three sows through AI using semen from one MSTN−/− boar. However, the genotype of piglets has not been tested at present. Thus, MSTN−/− boar may be used as sires, and these pigs are expected to be developed to provide new super-lean meat varieties in the future.

Plasma membrane changes during the liquid storage of boar spermatozoa: A comparison of methods

2010 ◽  
Vol 58 (1) ◽  
pp. 105-116 ◽  
Author(s):  
Dariusz Gączarzewicz ◽  
Małgorzata Piasecka ◽  
Jan Udała ◽  
Barbara Błaszczyk ◽  
Tomasz Stankiewicz ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Live Cells ◽  
Lower Percentage ◽  
Loss Of Function ◽  
Plasma Membrane Integrity ◽  
Sperm Plasma Membrane ◽  
Semen Preservation

Studies were performed on boar semen routinely used at the local artificial insemination (AI) centre. The semen was stored in a Safe Cell Plus commercial extender at 17 °C for nine days. The aim of our research was focused on changes in sperm plasma membrane integrity. The integrity of the sperm plasma membrane and acrosome as well as sperm motility decreased after dilution and during storage of the semen. The highest percentage of live sperm was identified by the eosin-nigrosin method, a lower percentage by the SYBR-14/PI test, and the lowest percentage of live cells was discovered by the hypoosmotic swelling (HOS) test (P < 0.01). There were significant differences between the results of staining methods and sperm motility (P < 0.01). No significant differences were found between the HOS test results and sperm motility. The plasma membrane integrity parameters positively correlated (P < 0.001) with each other and with sperm motility but negatively with aspartate aminotransferase activity. Our findings confirmed that the boar sperm aging changes, which increased during liquid semen preservation, were connected with the loss of function and integrity of the sperm plasma membrane. The employed complementary tests are comprehensive indicators of sperm membrane integrity during long-term semen preservation, and they can help establish the actual number of ‘healthy’ cells. The assays may be used in AI laboratories and should be incorporated into the routine of semen analysis.

scholarly journals Semen Quality of Garut Rams feed by Different Protein Sources and Their Implementation Potential in Small Farms of West Java

2020 ◽  
Vol 20 (1) ◽  
pp. 47-55
Author(s):  
Kurnia Bagus Ariyanto ◽  
Lilis Khotijah ◽  
Dewi Apri Astuti ◽  
Raden Iis Arifiantini ◽  
Jean-Baptise Menassol
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Black Soldier Fly ◽  
Hermetia Illucens ◽  
Brachiaria Humidicola ◽  
Plasma Membrane Integrity ◽  
Protein Sources ◽  
Sperm Plasma Membrane

ABSTRACT. Maggot Hermetia illucens (Maggot Black Soldier Fly, MBSF) is an alternative protein source besides soybean meal (SBM) which may be used as a feed for improving the quality of semen particularly in Garut rams to support prolific nature. The aims of this study were to analyzed and compare the impact of different protein sources in feed on semen quality of Garut rams, and to assess the prediction ability of Garut rams to serve ewe in small-scale breeders in West Java, Indonesia. This study was conducted using a completely randomized design with 3 treatments and 4 replications, consisted of Brachiaria humidicola (BH) grass and T1 (concentrate contains 20% of SBM), T2 (concentrate contains 10% of SBM and 10% of MBSF), and T3 (concentrate contains 20% of MBSF). The parameters measured were feed consumption, semen quality (macroscopic and microscopic characteristics), also a potential ability of rams to serve ewe. The results showed there were no significant effect on protein consumption, semen volume, semen pH, semen color and consistency, sperm mass movement, sperm motility, sperm concentration, sperm morphology, and prediction potential ability to serve ewe. However, the result showed a significant effect (P0.05) on sperm viability and sperm plasma membrane integrity. Sperm plasma membrane integrity of ram feed with T3 was better than T1 and T2 (P0.05). The prediction potential ability rams to serve ewes on MBSF treatment was 38 heads, while in T1 and T2 were 43 and 57 heads, respectively. In conclusion, MBSF can be an alternative source of protein besides SBM to improve the semen quality of Garut rams.  ABSTRAK. Maggot Hermetia illucens (Maggot Black Soldier Fly; MBSF) adalah sumber protein alternatif selain bungkil kedelai (SBM) yang dapat dipergunakan sebagai pakan untuk memperbaiki kualitas semen terutama pada domba Garut untuk mendukung sifat prolifik. Tujuan penelitian ini adalah menganalisis dan membandingkan dampak pemberian sumber protein berbeda terhadap kualitas semen domba Garut dan untuk menilai kemampuan domba Garut pejantan dalam melayani betina pada peternakan rakyat di Jawa Barat, Indonesia. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan 3 perlakuan dan 4 ulangan yang terdiri dari rumput Brachiaria humidicola (BH) dan T1 (konsentrat mengandung 20% SBM), T2 (konsentrat mengandung 10% SBM dan 10% MBSF), dan T3 (konsentrat mengandung 20% MBSF). Parameter yang diukur adalah konsumsi pakan, karakteristik semen (makroskopis dan mikroskopis) serta potensi domba jantan melayani betina. Hasil Penelitian menunjukkan tidak ada perbedaan signifikan pada konsumsi protein pakan, volume semen, pH semen, warna dan konsistensi semen, gerakan massa sperma, motilitas sperma, konsentrasi sperma, morfologi sperma, dan prediksi potensi pejantan dalam melayani betina. Namun, hasil penelitian menunjukkan terdapat perbedaan (P0.05) pada viabilitas sperma dan membran plasma utuh sperma. Membran plasma utuh pada perlakuan T3 lebih baik dibandingkan perlakuan T1 dan T2 (P0.05). Prediksi potensi betina terlayani dari pejantan yang diberi pakan MBSF adalah 38 ekor, sedangkan yang diberi SBM dan kombinasinya adalah 43 dan 57 ekor. Kesimpulan penelitian ini adalah MBSF dapat menjadi alternatif sumber protein selain bungkil kedelai dalam memperbaiki kualitas sperma domba Garut.

95 EFFECT OF CRYOPROTECTANT ADDITION AND REMOVAL AT 4°C ON MOTILITY AND PLASMA MEMBRANE INTEGRITY OF BOVINE CAUDAL EPDIDYMAL SPERMATOZOA

2006 ◽  
Vol 18 (2) ◽  
pp. 156
Author(s):  
C. Guerrero ◽  
S. Leibo ◽  
D. Paccamonti ◽  
B. Eilts ◽  
K. Bondioli ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Single Step ◽  
Freezing Process ◽  
Computer Assisted ◽  
Chemical Toxicity ◽  
Plasma Membrane Integrity ◽  
Epididymal Spermatozoa

Cryopreservation of spermatozoa harvested from the epididymides would be a means of salvaging germplasm from genetically valuable males that die unexpectedly from injury, disease, or poaching. It is well known that the addition of cryoprotective agents (CPAs) is essential for sperm survival following the freezing process. However, CPAs can cause loss in sperm viability due to osmotic damage or chemical toxicity. The objective of this study was to determine the effects of single-step addition and/or removal of glycerol (GLY) or ethylene glycol (EG) on motility and plasma membrane integrity of bovine epididymal spermatozoa. Paired testes were obtained from mature bulls (n = 10) at a local abattoir and transported to the laboratory at 25–28°C within 4–6 h post-mortem. Epididymal spermatozoa were harvested by multiple incisions from the caudae epididymides of each bull, pooled, and washed in Brackett-Oliphant medium by centrifugation for 5 min at 500g. Pellets were resuspended in egg yolk-Tris-glucose-citric acid monohydrate medium (EYT-GC) at a concentration of 120 × 106 cells/mL and cooled to 4°C at a rate of 0.1°C/min. Specimens were allocated to each of five treatment groups: control (no CPA), 7% GLY, and 14% GLY, 7% EG, 14% EG. Then, replicate samples were diluted 1:1 in EYT-GC medium containing twice the final desired concentration of CPA. After being exposed for 10 min, each sample was diluted directly into EYT-GC at 4°C. Motility was assessed by means of a computer assisted semen analysis system and plasma membrane integrity was determined by SYBR 14 and propidium iodide staining followed by fluorescence microscopy. Differences among treatments were analyzed using one way ANOVA (P < 0.05). The results (Table 1) show that maximum survival, as assessed by measurements of motility and membrane integrity, was achieved with spermatozoa exposed to 7% EG. Almost identical intermediate levels of survival were observed with spermatozoa exposed to 7% GLY or 14% EG. The lowest survival was observed for spermatozoa exposed to 14% GLY. The results indicate that the use of EG as a cryoprotectant may minimize toxicity and osmotic damage to fresh bovine epididymal spermatozoa. Its efficacy as a CPA is currently being determined. Table 1. Sperm motility and membrane integrity (mean ± SEM) after addition of CPA to epididymal sperm

78 DIFFERENT EXTENDERS TO HARVEST EQUINE EPIDIDYMAL SPERM AND THEIR INFLUENCE ON FREEZABILITY

2013 ◽  
Vol 25 (1) ◽  
pp. 186
Author(s):  
R. F. Soares ◽  
F. O. Papa ◽  
L. C. O. Magalhães ◽  
G. A. Monteiro ◽  
I. Martin ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Freezing Process ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Traumatic Injuries ◽  
Plasma Membrane Integrity ◽  
Quarter Horse ◽  
Pipette Tip

Harvesting and freezing epididymal sperm is a technology that enables the preservation of the gene pool from animals that had died either unexpectedly or because of colic conditions. This technique may also be employed in animals that have to be euthanized because of traumatic injuries. Therefore, the objective of the present study was to improve the process of freezing epididymal sperm using a freezing extender without the prior centrifugation of the samples. Twelve stallions aging between 3 and 6 years and of different breeds were used (Quarter Horse, Mangalarga, and Brazilian Jumping Horse). Stallions were castrated, and the cauda epididymides were isolated from the testis. The connective tissue was carefully dissected, and the cauda epididymides were straightened. A 200-µL pipette tip was attached to a 20-mL syringe, and the cauda epididymides were flushed using 40 mL of either (A) BotuSemen® (Nidacon, Mölndal, Sweden) or (B) BotuCrio® (Nidacon). They were then immediately processed at room temperature (25°C). Samples flushed with B were randomly subjected to either of the 2 procedures: B1) directly loaded into 0.5-mL straws or B2) centrifuged at 600g for 10 min, the supernatant was discarded, and the pellet was resuspended with B and loaded into 0.5-mL straws. The straws were kept at 5°C for 20 minutes followed by another 20 min at 6 cm above liquid nitrogen before immersion. After thawing at 46°C for 20 s the samples were analyzed by computer-assisted semen analysis (HTM – IVOS 12) and plasma membrane integrity was assessed using fluorescent probes (carboxyfluorescein diacetate and propidium iodide). Data were analyzed by ANOVA followed by the Tukey test (P < 0.05). No differences were observed for the values of total motility (A: 57.1 ± 12.35, B1: 46.3 ± 10.0, B2: 47.2 ± 12.84), progressive motility (A: 25.5 ± 9.05, B1: 21.7 ± 9.02, B2: 20.7 ± 7.20), percentage of rapids (A: 40.6 ± 15.92, B1: 30.7 ± 10.51, B2: 32.6 ± 12.39), and plasma membrane integrity (A: 47.8 ± 9.56, B1: 45.0 ± 13.81, B2: 41.3 ± 8.74). It was found that the fluid derived from epididymal secretions, which composes seminal plasma, had no influence on sperm parameters, because there was no difference among freezing protocols. Therefore, flushing equine epididymal cauda with B and freezing the samples without centrifuging can be successfully used. Both extenders (A and B) were efficient in protecting epididymal sperm throughout the freezing process.

Factors affecting storage of Slovak native rabbit semen in the gene bank

Zygote ◽  
2017 ◽  
Vol 25 (5) ◽  
pp. 592-600
Author(s):  
Barbora Kulíková ◽  
Marta Oravcová ◽  
Andrej Baláži ◽  
Peter Supuka ◽  
Peter Chrenek
Keyword(s):  
Plasma Membrane ◽  
Liquid Nitrogen ◽  
Semen Quality ◽  
Sperm Quality ◽  
Objective Assessment ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Quality Traits ◽  
Plasma Membrane Integrity

SummaryIn this study, fresh and frozen–thawed semen of Nitra and Zobor rabbit breeds were evaluated for potential inter-breed or inter-male differences in sperm quality traits. Individual male semen from four rabbits of each breed were diluted (v:v; 1:1) in a freezing medium composed of a commercial diluent, 16% of dimethyl sulphoxide (DMSO), 4% of Ficoll 70 and 2% of sucrose and frozen in liquid nitrogen vapours before being plunged into liquid nitrogen. Different motility traits, viability and plasma membrane integrity of fresh and frozen–thawed semen were evaluated in vitro using computer-assisted sperm analysis and flow cytometry. To evaluate the sperm fertilization ability, artificial insemination of fresh and frozen–thawed sperm was performed. Our results showed the effect of breed (P ≤ 0.05) on frozen–thawed sperm viability and plasma membrane integrity. Moreover, individual variability in semen quality among the rabbits was revealed (0.31 to 0.71 among quality traits). Our results thereby confirmed that the cryopreservation procedure could not ensure comparable sperm post-thaw survival for different breeds or males. Nevertheless, correlations between numbers of fresh total motile and progressively moving sperm and several quality parameters measured post thawing were revealed. Therefore, we suggest that the objective assessment of fresh rabbit sperm motility may be an effective indicator of frozen–thawed semen quality. Consequently, regular semen assessment is required in order to preserve good-quality insemination doses from native breeds.

Cryopreservation of collared peccary (Pecari tajacu L., 1758) epididymal sperm using extenders based on Tris and powdered coconut water (ACP®-116c)

Zygote ◽  
2018 ◽  
Vol 26 (4) ◽  
pp. 301-307 ◽  
Author(s):  
José A. B. Bezerra ◽  
Andréia M. Silva ◽  
Patrícia C Sousa ◽  
Lívia B. Campos ◽  
Érica C. G. Praxedes ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Coconut Water ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Sperm Plasma Membrane ◽  
Pecari Tajacu ◽  
Functional Membrane

SummaryThe aim of this study was to establish a functional freezing–thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen–thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

Immunolocalisation and expression of oxytocin receptors and sex hormone-binding globulin in the testis and epididymis of dogs: correlation with sperm function

2019 ◽  
Vol 31 (9) ◽  
pp. 1434
Author(s):  
Andressa Dalmazzo ◽  
João D. A. Losano ◽  
Daniel S. R. Angrimani ◽  
Isabel V. A. Pereira ◽  
Marcelo D. Goissis ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Protein Expression ◽  
Membrane Integrity ◽  
Sex Hormone ◽  
Sex Hormone Binding Globulin ◽  
Mitochondrial Activity ◽  
Hormone Binding ◽  
Plasma Membrane Integrity ◽  
Gene And Protein Expression ◽  
Acrosome Integrity

The aim of this study was to confirm gene and protein expression of oxytocin receptor (OTR) and sex hormone-binding globulin (SHBG) in the testis and epididymis of dogs, correlating these data with sperm quality and production and testosterone concentrations. Positive correlations were found between OTR and SHBG expression in both the testis and epididymis. Testicular OTR expression was positively associated with plasma membrane and acrosome integrity in canine spermatozoa, whereas SHBG expression in the testis was positively correlated with various sperm characteristics, such as sperm concentration, total and progressive motility, plasma membrane integrity and acrosome integrity. Testicular expression of both OTR and SHBG was negatively correlated with low sperm mitochondrial activity. In the epididymis, SHBG expression was only positively correlated with plasma membrane integrity. Analysis of protein expression revealed that testicular OTR was positively correlated with testosterone concentrations and negatively correlated with the absence of sperm mitochondrial activity. In addition, SHBG expression in the testes was associated with epididymis SHBG expression and morphologically normal cells. Immunohistochemical (IHC) analysis revealed the presence of both OTR and SHBG in testicular smooth muscles and Leydig cells. However, in the epididymis, OTR was only located in smooth muscle cells, whereas neither IHC nor western blotting detected SHBG. Together, the results of this study suggest that OTR and SHBG play key roles in spermatogenesis and sperm maturation, being essential for male reproductive success.

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