scholarly journals Vitamin C ameliorates tetrahydrocannabinol-induced spermatotoxicity in-vitro

2020 ◽  
Author(s):  
Abdullateef Isiaka Alagbonsi ◽  
Luqman Aribidesi Olayaki
Keyword(s):  
Vitamin C ◽  
Cannabinoid Receptors ◽  
Group Iv ◽  
Group Vi ◽  
Spermatozoa Motility ◽  
Progressive Motility ◽  
Straight Line ◽  
Lateral Head

Abstract Background: We investigated the in-vitro effects of vitamin C on delta-9-tetrahydrocannabinol (THC) -induced reduction in spermatozoa motility and kinematics. Methods: Six rats were used for the study. Semen from each of the 6 rats was randomly divided into 6 groups such that each rat’s semen was in all of the groups. Groups I-III received placebo, THC (1 mM), and vitamin C (5 mM) respectively. Group IV was pretreated with cannabinoid receptors’ blockers (CBs-) 1 and 2, followed by THC. Groups V and VI received THC and vitamin C, but group VI was additionally pre-treated with CBs-. Results: The spermatozoa progressive motility, average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), amplitude of lateral head (ALH) and beat cross frequency (BCF) were reduced by THC (6.08±1.16%; 5.64±0.82 µm/s; 6.96±0.74 µm/s; 2.75±0.23 µm/s; 0.31±0.02 µm; and 0.78±0.08 Hz respectively) but increased by vitamin C (51.20±1.32 %; 17.90±0.21 µm/s; 25.11±0.96 µm/s; 8.80±0.27 µm/s; 0.75±0.01 µm; and 3.15±0.03 Hz respectively) when compared to control (39.72±0.38 %; 13.70±0.29 µm/s; 18.04±0.58 µm/s; 7.54±0.34 µm/s; 0.65±0.02 µm; and 2.79±0.01 Hz respectively). Vitamin C inhibited the THC-induced reduction in these parameters (37.36±0.73 %; 10.98±0.45 µm/s; 13.58±0.30 µm/s; 7.11±0.22 µm/s; 0.58±0.01 µm; and 2.60±0.01 Hz respectively) in the absence of CBs- 1 and 2, and even caused additional increases in progressive motility (49.54±1.01 %), VAP (15.70±0.38 µm/s) and VCL (22.53±0.29 µm/s) above the control levels with CBs-.Conclusion: Vitamin C ameliorates the THC-induced reduction in spermatozoa motility in-vitro by modulation of their kinematics.

scholarly journals Vitamin C ameliorates tetrahydrocannabinol-induced spermatotoxicity in-vitro

2020 ◽  
Author(s):  
Abdullateef Isiaka Alagbonsi ◽  
Luqman Aribidesi Olayaki
Keyword(s):  
Vitamin C ◽  
Cannabinoid Receptors ◽  
Group Iv ◽  
Group Vi ◽  
Spermatozoa Motility ◽  
Progressive Motility ◽  
Straight Line ◽  
Lateral Head

Abstract Background: We investigated the in-vitro effects of vitamin C on delta-9-tetrahydrocannabinol (THC)-induced reduction in spermatozoa motility and kinematics. Methods: Six rats were used for the study. Semen from each of the 6 rats was randomly divided into 6 groups such that each rat’s semen was in all of the groups. Groups I-III received placebo, THC (1 mM), and vitamin C (5 mM) respectively. Group IV was pretreated with cannabinoid receptors’ blockers (CBs-) 1 and 2, followed by THC. Groups V and VI received THC and vitamin C, but group VI was additionally pre-treated with CBs-. Results: The spermatozoa progressive motility, average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), amplitude of lateral head (ALH) and beat cross frequency (BCF) were reduced by THC (6.08±1.16%; 5.64±0.82 µm/s; 6.96±0.74 µm/s; 2.75±0.23 µm/s; 0.31±0.02 µm; and 0.78±0.08 Hz respectively) but increased by vitamin C (51.20±1.32 %; 17.90±0.21 µm/s; 25.11±0.96 µm/s; 8.80±0.27 µm/s; 0.75±0.01 µm; and 3.15±0.03 Hz respectively) when compared to control (39.72±0.38 %; 13.70±0.29 µm/s; 18.04±0.58 µm/s; 7.54±0.34 µm/s; 0.65±0.02 µm; and 2.79±0.01 Hz respectively). Vitamin C inhibited the THC-induced reduction in these parameters (37.36±0.73 %; 10.98±0.45 µm/s; 13.58±0.30 µm/s; 7.11±0.22 µm/s; 0.58±0.01 µm; and 2.60±0.01 Hz respectively) in the absence of CBs- 1 and 2, and even caused additional increases in progressive motility (49.54±1.01 %), VAP (15.70±0.38 µm/s) and VCL (22.53±0.29 µm/s) above the control levels with CBs-.Conclusion: Vitamin C ameliorates the THC-induced reduction in spermatozoa motility in-vitro by modulation of their kinematics.

scholarly journals Vitamin C ameliorates tetrahydrocannabinol-induced spermatotoxicity in-vitro

BMC Nutrition ◽  
2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Abdullateef Isiaka Alagbonsi ◽  
Luqman Aribidesi Olayaki
Keyword(s):  
Vitamin C ◽  
Cannabinoid Receptors ◽  
Group Iv ◽  
Group Vi ◽  
Spermatozoa Motility ◽  
Progressive Motility ◽  
Straight Line ◽  
In Vitro Effects ◽  
Lateral Head

Abstract Background We investigated the in-vitro effects of vitamin C on delta-9-tetrahydrocannabinol (THC) -induced reduction in spermatozoa motility and kinematics. Methods Six rats were used for the study. Semen from each of the 6 rats was randomly divided into 6 groups such that each rat’s semen was in all of the groups. Groups I-III received placebo, THC (1 mM), and vitamin C (5 mM) respectively. Group IV was pre-treated with cannabinoid receptors’ blockers (CBs−) 1 and 2, followed by THC. Groups V and VI received THC and vitamin C, but group VI was additionally pre-treated with CBs−. Results The spermatozoa progressive motility, average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), amplitude of lateral head (ALH) and beat cross frequency (BCF) were reduced by THC (6.08 ± 1.16%; 5.64 ± 0.82 μm/s; 6.96 ± 0.74 μm/s; 2.75 ± 0.23 μm/s; 0.31 ± 0.02 μm; and 0.78 ± 0.08 Hz respectively) but increased by vitamin C (51.20 ± 1.32%; 17.90 ± 0.21 μm/s; 25.11 ± 0.96 μm/s; 8.80 ± 0.27 μm/s; 0.75 ± 0.01 μm; and 3.15 ± 0.03 Hz respectively) when compared to control (39.72 ± 0.38%; 13.70 ± 0.29 μm/s; 18.04 ± 0.58 μm/s; 7.54 ± 0.34 μm/s; 0.65 ± 0.02 μm; and 2.79 ± 0.01 Hz respectively). Vitamin C inhibited the THC-induced reduction in these parameters (37.36 ± 0.73%; 10.98 ± 0.45 μm/s; 13.58 ± 0.30 μm/s; 7.11 ± 0.22 μm/s; 0.58 ± 0.01 μm; and 2.60 ± 0.01 Hz respectively) in the absence of CBs− 1 and 2, and even caused additional increases in progressive motility (49.54 ± 1.01%), VAP (15.70 ± 0.38 μm/s) and VCL (22.53 ± 0.29 μm/s) above the control levels with CBs−. Conclusion Vitamin C ameliorates the THC-induced reduction in spermatozoa motility in-vitro by modulation of their kinematics.

scholarly journals Vitamin C Ameliorates Tetrahydrocannabinol-Induced Spermatotoxicity In-Vitro

2020 ◽  
Author(s):  
Abdullateef Isiaka Alagbonsi ◽  
Luqman Aribidesi Olayaki
Keyword(s):  
Vitamin C ◽  
Sperm Motility ◽  
Cannabinoid Receptors ◽  
Group V ◽  
Progressive Motility ◽  
Straight Line ◽  
Lateral Head

Abstract Background: We investigated the in-vitro effects of vitamin C on delta-9-tetrahydrocannabinol (THC)-induced reduction in sperm motility and kinematics. Methods: Rats semen were randomly divided into 5 groups. Groups I-III received placebo, THC (1 mM), and vitamin C (5 mM) respectively. Groups IV and V received THC and vitamin C, but group V was additionally pre-treated with cannabinoid receptors’ blockers (CBs-) 1 (SR141716) and 2 (AM-630). Results: The sperm progressive motility, average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), amplitude of lateral head (ALH) and beat cross frequency (BCF) were reduced by THC but increased by vitamin C when compared to control. Vitamin C inhibited the THC-induced reduction in these parameters in the absence of CBs- 1 and 2, and even caused additional increases in progressive motility, VAP and VCL above the control levels with CBs-. Conclusion: In conclusion, vitamin C ameliorates THC-induced reduction in sperm motility in-vitro by modulation of sperm kinematics.

scholarly journals Estimation of vitamin C human protective dose for acetaminophen toxicity, using acute animal toxicity study

2017 ◽  
Vol 2 (2) ◽  
pp. 101-105 ◽  
Author(s):  
Mudhaffar I. Al-Farras ◽  
Khalid K. Abdul-Razzak ◽  
Mohammed Yacoub ◽  
Wael M. Hananeh
Keyword(s):  
Mortality Rate ◽  
Vitamin C ◽  
Lethal Dose ◽  
Group Iv ◽  
Group Vi ◽  
Group V ◽  
Liver Histopathology ◽  
Toxic Damage ◽  
Group I ◽  
Group Ii

Acetaminophen (APAP) is the main cause of fulminant hepatic failure. Vitamin C is a natural antioxidant with protective potentials against APAP toxic damage. In this animal study, and after an LD50 determination and selection of suitable lethal dose, the investigation was done to select a proper protective dose of vitamin C against lethal APAP dose. All 6 animal groups received a lethal dose of APAP (3250 mg/kg), group II, III and IV received 500, 1000 and 1500 mg/kg vitamin C respectively, group V received 1200 mg/kg N-Acetylcysteine (NAC), and group VI receive 1000 mg/kg vitamin C and 1200 mg/kg NAC. Mortality was recorded and liver histopathology was carried out. The results showed, the mortality rate in the group I was 68.75% and 37.5%, 31.25% in group II and III respectively, while group IV Showed a higher mortality rate and in group V and VI it was 25%. There was also a gradual reduction in the grade of histopathological damage in all groups, ranging from 2.4 ± 0.55 in group I to 0.4 ± 0.55 in group V and VI. In conclusion, vitamin C showed an increasing reduction in mortality and more histopathological protection, and it was more significant at 1000 mg/kg. NAC adds no more protection or reduction in mortality. The estimated protective dose of vitamin C was 700 to1127 mg for each gram of APAP. Incorporation of this dose of vitamin C with APAP preparations may be considered as a promising method for reducing mortality or severity of APAP intoxication.

scholarly journals Efecto de la composición racial sobre la calidad espermática de verracos

2018 ◽  
Vol 29 (3) ◽  
pp. 485 ◽  
Author(s):  
Anthony Valverde-Abarca ◽  
Mónica Madrigal-Valverde ◽  
Marlen Camacho-Calvo ◽  
Adones Zambrana-Jiménez ◽  
Leonardo López
Keyword(s):  
Quality Evaluation ◽  
Semen Quality ◽  
Sperm Quality ◽  
Active Movement ◽  
Racial Groups ◽  
Swine Farms ◽  
Progressive Motility ◽  
Head Displacement ◽  
Straight Line ◽  
Lateral Head

The assessment of the semen quality is an essential tool to improve the reproductive indexes in swine farms. The motility is the most important parameter that used in the sperm quality evaluation because it is associated with the energy of the spermatozoon. The aim of this research was to determine the influence of breed composition on reproductive variables of sperm quality, kinetics and semen motility in boars. During 2016, 240 ejaculates were collected from 63 boars with a mean age of 24.4±10.9 months. Six racial groups were identified: Duroc (D), Yorkshire (Y), Landrace (L), F1 Pietrain*Duroc (PD) and two genetic lines (LA and LB). The breeds with the highest ejaculate volume presented a higher total number of spermatozoa (P<0.05). The effect of the breed was significant (P<0.05) on sperm kinetics variables except for amplitude of lateral head displacement (ALH, μm). The Landrace breed presented the highest (P<0.05) percentage of static sperm (29.30±1.57). The most relevant differences (P<0.05) for total motility (MTOT) and progressive motility (MP), were presented between L and PD with values of 70.71±1.57; 77.48±1.09 and 51.80±1.97; 59.85±1.37% respectively. Adult boars (≥18 months) had higher volumes of ejaculate and total number of sperm than boars in the intermediate and young ages, however, for velocities (μm/s): curvilinear (VCL), straight line (VSL) and average path (VAP), adult boars only were different (P<0.05) of the boars in-between ages. Four sperm subpopulations (SP) were identified, SP1 (46.83%) characterized by a moderate speed, but of very progressive motility, SP2 (14.78%) with active movement, but not progressive, SP3 (8.45%) with low speeds and progressive path and SP4 (29.94%) with quick movement, but without progressivity.

scholarly journals Effect of three commercial extenders on sperm motility and fertility in liquid ram semen stored at 15 °C or 5 °C

2019 ◽  
Vol 67 (3) ◽  
pp. 430-444
Author(s):  
Ander Arando ◽  
Juan Vicente Delgado ◽  
José Manuel León ◽  
Sergio Nogales ◽  
Francisco Javier Navas-González ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Soybean Lecithin ◽  
Egg Yolk ◽  
Kinematic Parameters ◽  
Progressive Motility ◽  
Head Displacement ◽  
Liquid Storage ◽  
Lateral Head

The effect of different extenders on sperm motility and fertility was evaluated during liquid storage of ram semen at 5 °C and 15 °C. The semen was collected, pooled and diluted in three commercial extenders: Inra 96® (INRA) based on skimmed milk, Biladyl® A fraction (BIL) based on egg yolk, and Ovixcell® (OVIX) based on soybean lecithin. Then, sperm motility was evaluated at 0, 6, 24, 48, 72 and 96 h. In order to evaluate fertility, samples stored at 15 °C were used after dilution in INRA and OVIX. Results showed that progressive motility was significantly higher up to 72 h of storage in sperm samples maintained at 5 °C in comparison with 15 °C, similarly for each tested diluent. When samples were stored at 5 °C in OVIX, kinematic parameters such as velocity (except curvilinear velocity, VCL), trajectory [linearity (LIN), straightness (STR), wobble (WOB)], amplitude of lateral head displacement (ALH) and beat/cross frequency (BCF) were higher than in INRA and BIL. No significant differences in pregnancy rate were detected between INRA (62.6%) and OVIX (58.9%). In conclusion, liquid storage at 5 °C with OVIX extender is an interesting option since non-animal components are used, and this extender offers similar in vitro and in vivo efficacy as other extenders containing animal components.

scholarly journals The The Influence of Wharton Jelly Mesenchymal Stem Cell toward Matrix Metalloproteinase-13 and RELA Synoviocyte Gene Expression on Osteoarthritis

2019 ◽  
Vol 7 (5) ◽  
pp. 701-706 ◽  
Author(s):  
Vivi Sofia ◽  
Ellyza Nasrul ◽  
Menkher Manjas ◽  
Gusti Revilla
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Inflammatory Responses ◽  
Group Iv ◽  
Group Vi ◽  
Group V ◽  
Group Iii ◽  
Group I

BACKGROUND: Therapy for osteoarthritis (OA) with satisfactory results has not been found to date. In OA pathogenesis, RELA gene involved in cartilage degradation and MMP-13 in degrade cartilage, as a member family of NF-ĸβ genes, RELA serves to modulate inflammatory responses and activates pro-inflammatory cytokines. AIM: This study aims to identify the influence of Wharton Jelly Mesenchymal Stem Cell (MSC-WJ) on MMP-13 and RELA expression gene in synoviocyte by in vitro. MATERIAL AND METHODS: This research is pure experimental research. The sample used derived from synovial tissue of OA patients who underwent Total Knee Replacement (TKR) surgery. This study was divided into six groups treated with 4 replications. Group I and II (control groups) were synoviocyte of OA incubated for 24 and 48 hours, respectively. Group III and IV were MSC-WJ incubated for 24 and 48 hours, respectively. Group V and VI were Synoviocyte-MSC-WJ co-culture group incubated for 24 and 48 hours, respectively. Identification of MMP-13 and RELA gene expression in each group was performed by using qPCR. RESULT: The results showed that MSC-WJ reduced MMP-13 gene expression after co-culture for 24 and 48 hours in OA synoviocyte. The highest gene expression of MMP-13 was in Group I and II (1.00 ng/μl), followed by Group III (0.41 ng/μl), Group IV (0.24 ng/μl), Group V (0.13 ng/μl), and Group VI (0.04 ng/μl). MSC-WJ administration also decreased RELA gene expression. The highest gene expression of RELA gene was in Group I and II (1.00 ng/μl), Group V (0.67 ng/μl), Group III (0.58 ng/μl), Group IV (0.16 ng/μl), and Group VI (0.16 ng/μl). CONCLUSION: This study concluded that MSC-WJ in OA synoviocyte significantly reduced the expression of MMP-13 and RELA gene (p <0.05).

scholarly journals Sorting and cryopreservation of goat sperm with or without phenolic compounds

2020 ◽  
Vol 72 (2) ◽  
pp. 295-304
Author(s):  
E.C.B. Silva ◽  
J.I.T. Vieira ◽  
I.H.A.V. Nery ◽  
R.A.J. Araújo Silva ◽  
V.F.M.H. Lima ◽  
...  
Keyword(s):  
Membrane Integrity ◽  
Living Cells ◽  
Phenolic Antioxidants ◽  
Density Gradients ◽  
Continuous Density ◽  
Progressive Motility ◽  
Head Displacement ◽  
Straight Line ◽  
Sperm Sorting ◽  
Lateral Head

ABSTRACT The objectives of this study were to evaluate goat sperm sorting in continuous Percoll® density gradients and gamete freezability, in the presence or absence of phenolic antioxidants. For this, semen pools were sorted, frozen, and evaluated. The non-selected group (NSg) presented lower progressive motility (PM), linearity (LIN), straightness (STR), and wobble (WOB) than the selected groups, and straight line velocity (VSL) compared to those with catechin or resveratrol. The amplitude of lateral head displacement (ALH) was higher in NSg, and quercetin reduced the mitochondrial membrane potential (MMP). After thawing, the NSg presented lower PM than the selected groups, VSL and VAP (average path velocity) than the selected group with or without catechin, LIN and WOB than the selected with or without catechin or resveratrol, and STR than the selected with catechin. Moreover, NSg presented higher ALH and BCF than the samples selected with or without catechin. Plasma membrane integrity and intact and living cells were higher in the selected groups, and MMP was lower in the NSg and the selected group with quercetin. Thus, centrifugation in Percoll® continuous density gradients is a viable methodology to select goat sperm compatible with the freezing, especially in the presence of catechin or resveratrol.

scholarly journals Mouse sperm delivery via cauda epididymis without transport medium

2016 ◽  
Author(s):  
Man-Xi Jiang ◽  
Mei-Shan Wang ◽  
Xiang-Hong Ou ◽  
Xue-Jin Chen ◽  
Yan Zhu
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Room Temperature ◽  
Sperm Quality ◽  
Mouse Sperm ◽  
Spermatozoa Motility ◽  
Transport Medium ◽  
Fertilization Rates ◽  
Progressive Motility

This study was aimed to investigate the effects of room temperature (RT, 20-25 oC) and absence of medium during cauda epididymis transport on spermatozoa quality, fertility and embryo development. In the first experiment, fresh sperm from one side of cauda epididymis was used for in vitro fertilization, and another side was delivered at RT or 4-8 oC either with or without M2. In the second experiment, each side of cauda epididymis obtained from the same mouse was individually delivered at RT or 4 oC with or without M2. Finally, sperm motility, progressive motility scores and fertility of fresh spermatozoa or those from transported cauda epididymis, and IVF embryo development were evaluated. Progressive motility scores and fertilization rates were higher in fresh spermatozoa than transported sperm; sperm motility of transported cauda epididymis at 4-8 oC was comparable to fresh spermatozoa, but spermatozoa motility of transported cauda epididymis at RT was inferior to fresh spermatozoa. Spermatozoa motillty of transported cauda epididymis at 4-8 oC with transport medium was much higher than that without transport medium; absence of transport medium did not affect sperm motility of transported cauda epididymis at 4-8 oC but affected sperm motility of transported cauda epididymis at RT. Sperm quality from transported cauda epididymis can be efficiently kept at 4-8 oC, and cauda epididymis transport at 4-8 oC without M2 is more beneficial on keeping their fertility. Moreover cauda epididymis transport at RT without medium could sufficiently produce embryos for obtainning live offsprings.

9 Single Layer Centrifugation of Bull Semen Through Percoll Plus® Before Cryopreservation

2018 ◽  
Vol 30 (1) ◽  
pp. 144
Author(s):  
A. Martins ◽  
F. N. Marqui ◽  
T. E. Cruz ◽  
T. I. H. Berton ◽  
D. G. Souza ◽  
...  
Keyword(s):  
Semen Quality ◽  
Single Layer ◽  
Computer Assisted ◽  
Treatment Groups ◽  
Progressive Motility ◽  
Bull Semen ◽  
Head Displacement ◽  
Straight Line ◽  
Artificial Vagina ◽  
Lateral Head

We previously reported that single layer centrifugation (SLC) with Percoll® (GE Healthcare, Uppsala, Sweden) of fresh bovine semen resulted in improved sperm progressive motility and movement, as evidenced by computer-assisted sperm analyzer (CASA) after freezing-thawing. However, no report has been found in the literature on the use of Percoll Plus® (PP; GE Healthcare), a nontoxic colloid, for the same purpose. Thus, this study aimed to verify the effects of SLC-PP before bull sperm freezing on sperm kinematics after cryopreservation. Ejaculates were collected from 3 Nellore bulls (6 from each) using an artificial vagina. After collection, the semen was assessed and pooled, and then 1 billion spermatozoa either diluted [D; 1:2 (v/v)] in freezing extender (FE, without glycerol) or undiluted (UD) was layered on top of a 9-mL column of PP (in 15-mL centrifuge tubes) at concentrations of 70% or 90% to form the 70D, 70UD, 90D, and 90UD treatment groups. Following centrifugation for 13 min at 839 × g [except for the control (C) group], the supernatant was removed and the sperm pellet diluted to 50 × 106 sperm mL−1 in FE medium plus glycerol. Then, frozen–thawed sperm samples were analysed by CASA (MMC Sperm, St. Petersburg, Russia) for the following parameters: total motility (TM, %), progressive motility (PM, %), curvilinear velocity (VCL, µm−1), straight line velocity (VSL, µm s−1), average path velocity (VAP, µm s−1), amplitude of lateral head displacement (ALH, µm), beat cross frequency (BCF, Hz), linearity (LIN, %), and straightness (STR, %). For statistical analyses, ANOVA and Student-Newman-Keuls test were used. Data are presented as mean ± SEM with P < 0.05 taken as significant. No difference was found among the groups for TM, VSL, BCF, and STR. However, the percentage of PM was higher (P < 0.05) in the SLC-selected sperm samples (values ranging from 42.0 ± 7.0 to 47.4 ± 11.4) than in C (28.8 ± 5.0), and ALH was lower in 70UD (1.6 ± 0.12) and 70D (1.7 ± 0.10) than in C (1.9 ± 0.2). Moreover, 70UD (49.0 ± 1.0), 90UD (50.0 ± 3.0), and 90D (50.0 ± 4.0) displayed higher percentage of LIN (P < 0.05) compared with C (45.0 ± 2.0) and 70D (48.0 ± 3.0). On the other hand, similar results were obtained for VCL (from 126.3 ± 8.0 to 130.0 ± 20.5) and VAP (from 82.7 ± 14.5 to 85.1 ± 6.9) in C, 70UD, and 70D, but these values differed (P < 0.05) from those for VCL in 90UD (104.6 ± 10.3) and 90D (97.2 ± 22.0) as well as for VAP in 90UD (72.2 ± 11.0) and 90D (71.8 ± 9.6). These are the first data demonstrating favourable influences of SLC with 70% Percoll Plus® to select distinct sperm subpopulations as evidenced by enhanced PM, LIN, and ALH. Thus, SLC-PP could optimize the production of frozen bull semen by decreasing the number of sperm per insemination dose, and help to circumvent limitations associated with the poor semen quality sometimes found in bulls of high genetic merit. This research was funded by FAPESP # 2015/20986-3, MasterFertility and Tairana Artificial Insemination Station, Brazil.

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