Computer assisted sperm analysis: Relationship between the movement characteristics of buffalo spermatozoa and sire fertility

Author(s):  
Raushan K Singh ◽  
A. Kumaresan ◽  
M. A. Mir ◽  
P. Kumar ◽  
S. Chhillar ◽  
...  

The present study was undertaken to identify the differences in sperm kinematics between buffalo bulls with different fertility ratings. Murrah buffalo bulls (n=9) that were routinely used for breeding purpose under progeny testing programme were utilized for the study. Bull fertility was determined based on in vivo fertility trials and the conception rates (CR) were adjusted for different non-genetic parameters. Based on the adjusted CR, bulls were classified into high, medium and low fertile group. Frozen semen samples of these bulls were obtained and sperm kinematic parameters were assessed using a computer assisted sperm analyzer. The kinematic parameters analyzed included the curvilinear velocity (VCL), the linear velocity (VSL), the average path velocity (VAP), the amplitude of lateral head displacement (ALH), the linearity (LIN), the straightness coefficient (STR) and the beat cross frequency (BCF). In high fertile bulls, the proportion of motile spermatozoa was higher (p<0.001) than the medium and low fertile bulls. The VAP and VCL of sperm motion were significantly higher (P<0.05) in high fertile bulls compared to either medium or low fertile bulls. The VSL was significantly lower in low fertile bulls (P<0.005) compared to either high or medium fertile bulls. Spermatozoa from high fertile bulls had significantly higher (P<0.05) BCF, STR, ALH and LIN compared to either medium or low fertile bulls. Buffalo bull fertility was significantly and positively correlated with sperm motility, VAP, VSL, VCL and ALH.

2019 ◽  
Vol 67 (3) ◽  
pp. 430-444
Author(s):  
Ander Arando ◽  
Juan Vicente Delgado ◽  
José Manuel León ◽  
Sergio Nogales ◽  
Francisco Javier Navas-González ◽  
...  

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.


2013 ◽  
Vol 25 (1) ◽  
pp. 183 ◽  
Author(s):  
I. Ortiz ◽  
J. Dorado ◽  
D. Acha ◽  
L. Ramirez ◽  
M. Urbano ◽  
...  

Single-layer centrifugation (SLC) with EquipureTM Bottom Layer has been used to enhance the quality of stallion semen samples; however, no studies have been performed on donkeys. The aim of this study was to determine if SLC with EquipureTM Bottom Layer improves kinematic parameters on frozen–thawed donkey sperm. Semen was collected from 4 Andalusian donkeys by artificial vagina. Three ejaculates from each donkey were centrifuged with EquiproTM, supernatant was removed, and pellet was re-extended in the freezing medium GentTM to a final concentration of 200 × 106 spermatozoa per milliliter. Sperm were slowly cooled to 5°C for 2 h, loaded in 0.5-mL plastic straws, and frozen in liquid-nitrogen vapors. After at least one week of storage, straws were thawed in a water bath at 37°C for 30 s. After thawing, semen samples were divided in 2 aliquots: aliquot 1 was used as such (control) and aliquot 2 was processed by SLC using EquipureTM Bottom Layer. Computer-assisted sperm analysis was performed, and sperm kinematics total motility (%), progressive motility (%), curvilinear velocity (VCL; µm s–1), velocity straight line (VSL; µm s–1), velocity average path (VAP; µm s–1), linearity (LIN; %), straightness (STR; %), wobble (WOB; %), lateral head displacement (ALH; µm), and beat cross frequency (BCF; Hz) were statistically compared using GLM model between frozen–thawed semen samples processed or not with EquipureTM. Results were expressed as mean ± standard error. Significant differences (P < 0.05) were found between SLC-selected and unselected semen for total motility (77.44 ± 5.83 v. 58.89 ± 6.07), progressive motility (76.88 ± 4.52 v. 56.59 ± 5.44), VCL (137.50 ± 0.75 v. 133.0 ± 0.99), LIN (69.43 ± 0.31 v. 68.23 ± 0.41), STR (78.45 ± 0.29 v. 76.90 ± 0.37), WOB (85.06 ± 0.18 v. 83.91 ± 0.26), ALH (2.76 ± 0.01 v. 2.44 ± 0.01), and BCF (9.13 ± 0.05 v. 8.53 ± 0.06), respectively. No significant differences were observed for VSL (102.89 ± 0.70 v. 104.32 ± 0.95) and VAP (123.21 ± 0.71 v. 121.50 ± 0.98). Most of the computer-assisted sperm analysis parameters used in the present study have been previously identified as reliable markers of sperm motility in relation to sperm quality and fertility. It has also been reported that VCL appears to be critical for the formation of the sperm reservoir and penetration of the zona pellucida. In addition, other variables improved in the SLC-selected samples have been described as measure of progressivity (LIN, STR) and spermatozoa vigor (BCF, ALH). These preliminary results suggest an additional option for improving sperm quality in donkey semen doses. In conclusion, SLC with EquipureTM can be used to enhance kinematic parameters on frozen–thawed donkey sperm.


2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
P. Perumal ◽  
S. K. Srivastava ◽  
S. K. Ghosh ◽  
K. K. Baruah

The present study was undertaken to assess the motility and velocity parameters of sperm of freezable and nonfreezable ejaculates by computer-assisted sperm analyser (CASA) such as Hamilton-Thorne Semen Analyser IVOS 11 in mithun semen. Fifty ejaculates (twenty-five ejaculates each for freezable and nonfreezable semen ejaculates) were collected from ten matured mithun bulls. CASA parameters, motility parameters such as forward progressive motility (FPM) (%), nonprogressive motility (NPM) (%), total motility (TM) (%), and static sperms (SM) (%); velocity parameters such as curvilinear velocity (VCL) (μm/sec), straight line velocity (VSL) (μm/sec), average path velocity (VAP) (μm/sec), linearity (LIN) (%), straightness (STR) (%), wobble (WOB) (%), amplitude of lateral head displacement (ALH) (μm), and beat/cross-frequency (BCF) (Hz) were measured by CASA analyser. The result revealed that these parameters varied significantly (P<0.05) between the freezable and nonfreezable ejaculates and freezable ejaculates have significantly (P<0.05) higher value than nonfreezable ejaculates. It was concluded that most of the CASA parameters were significantly lower in nonfreezable ejaculates than in freezable ejaculates in mithun and confirmed that the CASA was effective for a quick and objective analysis of motility and velocity parameters in mithun semen.


1994 ◽  
Vol 61 (1) ◽  
pp. 75-80
Author(s):  
G. Bonanni ◽  
A. Colonna ◽  
S. Masiero ◽  
I. Mastrogiacomo

The recent introduction of various devices for computer-assisted sperm analysis allows us to have not only objective measurements of the seminal parameters but also to determine some characteristics of sperm motion such as curvilinear and linear velocity and amplitude of lateral head displacement that cannot be obtained by routine analysis. These new seminal parameters seem to be very useful in the diagnosis of male infertility and, among them, ALH seems to play a more important role; in one of our studies we found a significant correlation between this parameter and the results of the bovine cervical mucus penetration test (Penetrak). Furthermore it is possible to determine sperm subpopulations for each parameter and so it will be possible to express a more accurate judgement not only on the fertilizing capacity of semen but also to know the effects on spermatozoa of different activating and capacitating treatments.


Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1993
Author(s):  
Sabrina Gacem ◽  
Jaime Catalán ◽  
Anthony Valverde ◽  
Carles Soler ◽  
Jordi Miró

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 106 sperm/mL offers the best choice for donkey computerised sperm motility analysis.


2009 ◽  
Vol 21 (9) ◽  
pp. 102
Author(s):  
S. M. H. Andrabi ◽  
M. Shahab

The present study was designed to investigate the bacterial species incriminated in bubaline semen and to find out the effectiveness of antibiotics (GTLS; gentamycin, tylosin and linco-spectin or SP; streptomycin and penicillin) in cryodiluent on bacterial control and quality of buffalo bull spermatozoa. For this purpose four experiments were conducted. In experiment 1, a total of 11 bacterial species were identified from buffalo ejaculates. The predominant bacteria were Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa in the ejaculates. In experiment 2, total aerobic bacterial counts in post-thaw samples were lower (P<0.05) in GTLS than in SP or NC (negative control). Fewer bacterial genera were identified in semen samples having GTLS than SP. Majority of the bacterial isolates from ejaculates showed more sensitivity towards GTLS than SP. In experiment 3, motilities (visual and computer-assisted), velocities (straight-line, average path and curvilinear), amplitude of lateral head displacement and plasma membrane integrity in post-thaw semen samples did not differ (P>0.05) due to antibiotics. Spermatozoal abnormalities (acrosome, head, mid-piece and tail) were lower (P<0.05) in GTLS and SP than in NC. In experiment 4, the fertility rates for SP-based vs. GTLS-containing frozen semen of buffalo bull were 42.8 and 55.2%, respectively. The results for GTLS were significantly higher than SP. The fertility rates also differed significantly in the first and second batch of inseminations performed with SP or GTLS-based cryopreserved semen of buffalo bull. In conclusion, a number of bacterial species are isolated from bubaline semen. Bacterial and seminal quality measured by standard laboratory tests and field fertility trials indicate that GTLS is more suitable in extender for cryopreservation of buffalo bull spermatozoa.


1994 ◽  
Vol 74 (1) ◽  
pp. 53-58 ◽  
Author(s):  
J. L. Bailey ◽  
M. M. Buhr ◽  
L. Robertson

Correlations among computer-assisted spermatozoa motility analyses, Ca2+ fluxes and in vivo fertility of bovine spermatozoa based on a total of 4482 inseminations were investigated in each of four ejaculates from six bulls. The Ca2+ parameters assessed the rate of change in intra- and extracellular Ca + in fresh and cryopreserved spermatozoa from the same ejaculates and were described in another study. Of the seven motility parameters of cryopreserved semen investigated, all differed significantly among bulls but none were related to the in vivo fertility of cryopreserved semen. The amplitude of lateral head displacement, a motility parameter associated with hyperactivation, was positively correlated with the intracellular Ca2+ levels and the rate of Ca2+ accumulation of cyropreserved spermatozoa. The highest fertility was observed when initial extracellular Ca2+ for cryopreserved spermatozoa was high and when Ca2+ efflux rates of cryopreserved cells approached the higher efflux rates of fresh spermatozoa. Fertility was reduced when cryopreserved spermatozoa had initial internal Ca2+ levels greater than those of fresh spermatozoa or when cryopreserved spermatozoa internalized Ca2+ rapidly. Calcium flux, but not motility, may predict fertilizing capacity of cryopreserved bovine spermatozoa. Key words: Calcium, computer-analysed motility, fertility, spermatozoa, bull


Reproduction ◽  
2005 ◽  
Vol 129 (1) ◽  
pp. 39-49 ◽  
Author(s):  
Maria Albrizio ◽  
Antonio Ciro Guaricci ◽  
Filippo Maritato ◽  
Raffaele Luigi Sciorsci ◽  
Gaetano Mari ◽  
...  

The development of fertilizing ability in sperm cells is associated with changes in the plasma membrane. However, to date the exact nature of sequentially activated primary receptors and channels and the signal transduction pathways derived from these remains elusive. We analyzed the expression and localization of the μ-opioid receptossr in equine spermatozoa. A transcript corresponding to the third extracellular loop that selectively binds μ agonists was amplified, sequenced and compared with the known sequences in humans, rats and cattle. The amplification product showed a high degree of nucleotide conservation. By immunofluorescence, μ-opioid receptor labeling was found on the sperm head and on the tail and disappeared in the acrosomal region of acrosome-reacted sperm cells. Immunoblotting revealed two bands of 50 and 65 kDa. Effects of the opioid antagonist naloxone on motility and on viability and capacitation/acrosome reaction were investigated by computer-assisted sperm analysis and Hoechst 33258/chlortetracycline (H258/CTC) staining. Progressive motility was significantly reduced after 3 h incubation in 10−3 M naloxone (P < 0.05), whereas it increased significantly after 5 h in 10−8 M naloxone (P < 0.05). Sperm velocity at 5 h was significantly reduced by the addition of 10−3 M naloxone (P < 0.05), but increased significantly in the presence of 10−8 M (P < 0.001). Curvilinear velocity and amplitude of lateral head displacement in spermatozoa incubated in the presence of naloxone were not indicative of hyperactivation. H258/CTC staining showed that 10−8 M naloxone significantly stimulated capacitation (P < 0.01) after 3 h. However, it had no effect on sperm cell viability and acrosomal status. Overall, this study provides the first evidence that the μ-opioid receptor is expressed in equine spermatozoa and that naloxone significantly affects motility and capacitation.


2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 115-115
Author(s):  
Carl R Dahlen ◽  
Sarah R Underdahl ◽  
Matthew S Crouse ◽  
Kacie L McCarthy ◽  
Cierrah J Kassetas ◽  
...  

Abstract Fifteen mature beef bulls (BW = 800.4 ± 17.4 kg) were used in a 112-d experiment to evaluate effects of divergent planes of nutrition on motility and kinematic properties of fresh and frozen-thawed semen. Bulls were ranked by BW and randomly assigned to one of two treatments: 1) managed on a positive plane of nutrition (POS, n = 8), or 2) managed on a negative plane of nutrition (NEG, n = 7). Bulls were fed a common diet, adjusted biweekly to achieve targeted weight loss or gain of 12.5% of original BW. On d 112, electroejaculation was used to collect 2 ejaculates from each bull, which were combined. An aliquot of fresh semen was evaluated via computer-assisted semen analysis (CASA; IVOS II, Hamilton Thorne, Beverly, MA, USA) for motility and kinematic properties. Remaining semen was extended and frozen. Frozen semen was thawed for 40 s and held in a heating block at 37°C, then evaluated via CASA at 0 and 3 h post-thaw. Data were analyzed in the MIXED procedure of SAS, with data for post-thaw analysis evaluated as repeated measures in time. Treatment did not influence ejaculate volume or concentration (P ≤ 0.19). In fresh ejaculates no impacts (P ≤ 0.29) of treatment were observed for motility or kinematic properties. In frozen-thawed ejaculates, however, bulls in the NEG treatment had greater (P ≤ 0.02) proportions of motile and progressively motile sperm compared with POS. In sperm classified as motile or progressively motile, NEG had greater (P ≤ 0.002) average path and straight line velocities, and greater (P ≤ 0.05) amplitude of lateral head displacement than POS. Treatment impacts observed in frozen, but not fresh, indicate that sperm metabolism, mitochondrial function, antioxidant capacity, or other factors may be influenced by plane of nutrition resulting in altered motility and kinematic properties.


Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1885
Author(s):  
José Néstor Caamaño ◽  
Carolina Tamargo ◽  
Inmaculada Parrilla ◽  
Felipe Martínez-Pastor ◽  
Lorena Padilla ◽  
...  

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.


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