scholarly journals Correlation between the sperm characteristics evaluated by a computerized system (CASA) and the reproductive performance of sows

2020 ◽  
Vol 9 (6) ◽  
pp. e09963389
Author(s):  
Emanuelle Maria Gottardi ◽  
Thaisa Campos Marques ◽  
Karen Martins Leão ◽  
Francisco Ribeiro de Araújo Neto ◽  
Leidiane Gonçalves Fernandes
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Head Displacement ◽  
Sperm Characteristics ◽  
Flagellar Beat ◽  
Straight Line ◽  
Computerized System ◽  
Farrowing Rate

Computer-assisted semen analysis (CASA) systems have been one the most used tools to evaluate sperm kinetics. The objective of this research was to estimate the correlation between sperm motility characteristics evaluated by CASA during 72 hours of cooling with the farrowing rate (FR) and total number of piglets born (TNB) after artificial insemination. Multiparous sows (n=464) were inseminated with semen from seven boars (19.6 ± 1.3 ejaculates/male). Sperm motility parameters were determined immediately after dilution and after 24, 48 and 72 hours of cooling at 15°C: total motility (TM-%), progressive motility (PM-%), curvilinear velocity (VCL-μm/s), straight line velocity (VSL-μm/s), average path velocity (VAP-µm/s), amplitude of lateral head displacement (ALH-µm), flagellar beat cross frequency (BCF-Hz), straightness (STR-%) and linearity (LIN-%). Pearson’s correlation coefficient was applied to analyze the data and the comparison of the means of the sperm characteristics between the boars was done by Tukey’s test at 5% probability. TM and PM at time zero (T0) were significant and had a moderate to high correlation with FR and TNB. After 72 hours of refrigeration, the semen quality was reduced and showed a significant and low correlation of the TM and PM with these same parameters. The boar presenting the lowest value of TM and PM after dilution obtained lower FR and TNB. In conclusion, computer-assisted semen analysis soon after dilution can be used to predict fertility of boars.

149 The second isoform of gonadotrophin-releasing hormone and its receptor affect boar semen quality

2020 ◽  
Vol 32 (2) ◽  
pp. 201
Author(s):  
C. E. Ross ◽  
F. H. Choat ◽  
K. N. Plager ◽  
A. T. Desaulniers ◽  
R. A. Cederberg ◽  
...  
Keyword(s):  
Semen Quality ◽  
Computer Assisted ◽  
Functional Protein ◽  
Littermate Control ◽  
Normal Morphology ◽  
Releasing Hormone ◽  
Sperm Analysis ◽  
Sperm Characteristics ◽  
Straight Line ◽  
Gonadotrophin Releasing Hormone

Pigs are the only livestock species encoding a functional protein for both the second isoform of gonadotrophin-releasing hormone (GnRH-II) and its cognate receptor (GnRHR-II). Unlike the classical GnRH system (GnRH-I and GnRHR-I), GnRH-II and GnRHR-II are abundantly produced in porcine testes. Moreover, GnRH-II binding its receptor on Leydig cells stimulates luteinizing hormone-independent testosterone secretion. Interestingly, GnRHR-II is also localised to the connecting piece of mature, ejaculated spermatozoa, whereas GnRH-II is detected in seminal plasma, an interaction possibly influencing the function of sperm. To examine the role of GnRH-II and its receptor in the testis, we produced a swine line with reduced endogenous GnRHR-II levels (GnRHR-II KD). The objectives of this study were to (1) compare sperm characteristics between mature GnRHR-II KD and littermate control boars on the day of collection and following semen extension and (2) determine whether a GnRHR-I and GnRHR-II antagonist alters sperm characteristics after storage of extended semen. In Experiment 1, GnRHR-II KD (n=3) and littermate control (n=3) ejaculates were collected (Day 1) and computer-assisted sperm analysis (CASA) was performed (IVOS II Animal; Hamilton Thorne) to determine measures of sperm motion (motility, progressive motility, slow, and static), morphology (normal morphology, bent tail, coiled tail, distal droplet, proximal droplet (PD), distal midpiece reflex, elongation, and area), and kinematics (length of average path (DAP), length of straight line path (DSL), length of curvilinear path (DCL), average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), straightness (STR), linearity (LIN), amplitude of lateral head displacement (ALH), beat-cross frequency, and wobble (WOB)). Next, 3 billion sperm were extended with Androstar Plus (80-mL doses; Minitube) and stored at 17°C until Day 7 CASA. Data were analysed with the MIXED procedure of SAS (SAS Institute Inc.). On Day 1, semen doses from GnRHR-II KD boars had reduced DSL, VSL, STR, LIN, and WOB (P<0.05), whereas sperm from control boars possessed more PD (P<0.01). Day 7 CASA revealed that transgenic sperm had reduced DAP, DCL, VAP, and VCL, although sperm from control boars were slower (P<0.05). In Experiment 2, control ejaculates (n=3) were extended as above, treated with increasing concentrations (0, 0.0001, 0.001, 0.01, 0.1, 1, and 10μM) of a GnRH antagonist inhibiting both GnRHR-I and GnRHR-II (SB-75, cetrorelix), and stored at 17°C until Day 7 and 9 CASA. On Day 7, only sperm characteristics in doses treated with 10μM SB-75 were significantly lower (normal morphology, DAP, DCL, VAP, VCL, and ALH) or higher (PD, WOB, and area) than controls. Similar differences (except ALH; P<0.10) for the 10μM SB-75 treatment were detected on Day 9; however, motility, slow, static, STR, and LIN were also reduced (P<0.05). Thus, these data suggest that GnRH-II and its receptor are important to sperm function, representing a potential avenue to improve semen preservation. This research was funded by USDA/NIFA AFRI (2017-67015-26508; BRW).

scholarly journals Seasonal variations in quantitative and qualitative sperm characteristics in fertile and subfertile stallions

2020 ◽  
Vol 63 (1) ◽  
pp. 145-154 ◽  
Author(s):  
Yara Suliman ◽  
Frank Becker ◽  
Armin Tuchscherer ◽  
Klaus Wimmers
Keyword(s):  
Breeding Season ◽  
Seasonal Variations ◽  
Semen Quality ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Quality Parameters ◽  
Computer Assisted ◽  
Normal Sperm ◽  
Straight Line ◽  
Significant Seasonal Variation

Abstract. Horses are seasonal breeders with a natural breeding season beginning in spring and extending through midsummer. In this study, quantitative and qualitative parameters of chilled stallion semen were compared between fertile and subfertile stallions and between the breeding and the non-breeding season. Semen quality parameters compared included ejaculate volume, sperm concentration, total sperm number, sperm morphology, and computer-assisted semen analysis (CASA)-derived sperm movement characteristics obtained from two groups of warmblood stallions (n=8; four fertile stallions and four subfertile stallions), which differ in the seasonal pregnancy rate 80 %–90 % (fertile) vs. 40 %–60 % (subfertile). A total of 64 ejaculates were collected from the stallions (n=8; four in the breeding season and four in the non-breeding season of each stallion). No significant differences in the semen quality parameters between the fertile and the subfertile stallions in the non-breeding season were observed. However, in the breeding season the proportion of morphologically normal sperm, total motility, progressive motility, average path velocity (VAP), and curvilinear velocity (VCL) were significantly higher in the fertile group (P<0.05) when compared with the subfertile group. In addition, a significant seasonal variation in the proportion of morphological normal sperm was found in the fertile group between the breeding and the non-breeding season (P<0.05). Moreover, significant seasonal variations (P<0.05) in CASA parameters of mean VAP, straight line velocity (VSL), and beat-cross frequency (BCF) were observed in the fertile and the subfertile stallions, which tended to be lower in the non-breeding season. In conclusion, differences between the fertile and the subfertile stallions were observed only in the breeding season, and a few of CASA-derived parameters seemed to be significantly lower during the non-breeding season in both the fertile and the subfertile stallions.

180 ASSESSMENT OF BULL SEMEN QUALITY LOADED IN NEW SensiTemp STRAWS USING SEMEN AND IN VITRO PRODUCTION TECHNOLOGIES

2016 ◽  
Vol 28 (2) ◽  
pp. 221
Author(s):  
D. Le Bourhis ◽  
S. Camugli ◽  
P. Salvetti ◽  
L. Schibler ◽  
E. Schmitt
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Cleavage Rate ◽  
Fluid Medium ◽  
And Control

SensiTemp, a new in vitro maturation (IMV) bull straw concept, presents the advantage of colour changing while the straw is thawed. The colour of frozen straws is blue and straws start to become white when the temperature reaches 33°C, with a complete change of colour at 37°C. The objective of this study is to assess sperm quality after thawing of semen frozen in SensiTemp from 2 bulls, by analysing, in experiment 1, sperm motility and membrane integrity using computer-assisted semen analysis (CASA) and flow cytometry (FC), and, in experiment 2, the in vitro embryo production (IVP) using IVP technologies [IVM, IVF, and in vitro culture (IVC)]. The ejaculates of 2 bulls, selected during preliminary experiments on high in vitro fertility, were harvested at CIA L’Aigle, France, and split ejaculates were frozen in experimental (SensiTemp) and conventional (control) straws. In experiment 1 after thawing semen from the 2 types of straws (5 pooled straws each; 2 replicates), motility was assessed using the IVOS CASA system (Hamilton Thorne Inc., Beverly, MA, USA) and membrane integrity was evaluated through FC with Cytosoft software (Millipore-Guava Technologies Inc., Hayward, CA, USA). In experiment 2, IVF was used to evaluate the non-toxicity of SensiTemp and control straws. Cumulus-oocyte complexes (COC; n = 1178; 4 replicates) collected from slaughterhouse ovaries were matured in IVM medium (TCM-199 with bicarbonate, Sigma-Aldrich, Saint Quentin Fallavier, France; 10 µg mL–1 FSH-LH, Reprobiol, Liège, Belgium; and 10% FCS, Thermo Fisher, Illkirch, France) for 22 h. After fertilization, presumptive zygotes of each group (SensiTemp and control for each bull) were cultured in synthetic oviduct fluid medium (SOF, Minitube, Tiefenbach, Germany) with 1% estrous cow serum (ECS) and 0.6% BSA (Sigma-Aldrich, France) up to 8 days. All cultures were conducted at 38.5C in 5% CO2, and 5% O2. The cleavage and blastocysts rates were evaluated on Days 3 and 7, respectively, for each group. Embryo quality was recorded on Day 7 according to the IETS evaluation. Data from each bull were analysed separately using the chi-squared test (P < 0.05). In experiment 1, neither sperm motility from bull 1 (61.2 and 60.5%) and bull 2 (66.2 and 66.5%) nor membrane integrity from bull 1 (58.6 and 52.2%) and bull 2 (61.0 and 61.9%) were different between SensiTemp and control, respectively. Results from experiment 2 showed no difference (P > 0.05) in cleavage rate between SensiTemp and control for the 2 bulls: 92.1 and 91.7% for bull 1 and 94.2 and 94.6% for bull 2 respectively. The blastocysts rate on Day 7 did not differ (P > 0.05) among groups (47.5, 47.1 and 51.3, 50.4% for SensiTemp and control bull 1 and bull 2, respectively) nor the quality of embryos retrieved in the different groups: 25.4, 23.3, and 30.8, 29.6% in grade 1 embryo for SensiTemp and control bull 1 and bull 2, respectively. Those results demonstrate, in vitro, that the new SensiTemp straws were non-toxic and did not affect the semen quality after thawing nor did the SensiTemp straws affect the ability of sperm cells to fertilize oocytes and produce 8-day-old embryos.

Computer assisted sperm analysis of fresh and frozen-thawed buffalo semen and their interrelationship

2016 ◽  
Vol 50 (1) ◽  
Author(s):  
J. B. Patel ◽  
A. J. Dhami
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Motile Sperm ◽  
Mean Values ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Straight Line ◽  
Buffalo Semen ◽  
Live Sperm ◽  
Fresh Semen

Sixty semen ejaculates from 10 mature bulls, 5 each of Jafarabadi and Mehsana breed, were studied for sperm motility and velocity parameters of fresh and frozen-thawed spermatozoa using computer assisted sperm analyzer (CASA). The mean values of motile and progressively motile spermatozoa observed in fresh semen of Jafarabadi and Mehsana bulls (79.77±1.62 and 61.80±1.85, and 78.90±1.22 and 61.37±1.58%) were highly significantly (P<0.01) reduced (51.20±1.57 and 33.20±1.45, and 52.10±1.70 and 34.30±1.54 %, respectively) in post-thawed semen. The average path velocity, straight line velocity and curvilinear velocity (μm/sec) of spermatozoa of Jafarabadi and Mehsana bulls noted in fresh semen were also reduced highly significantly (P<0.01) in frozen-thawed semen. Among the other velocity parameters, amplitude of lateral head displacement (μm), elongation (%) and medium motile sperm (%) increased, while beat-cross frequency (Hz), straightness (%), linearity (%), sperm area (μm<sup>2</sup>) and rapidly motile sperm (%) decreased significantly in post-thawed sperms when compared with the fresh sperm of both Jafarabadi and Mehsana bulls. The initial motility and live sperm per cent were significantly correlated with CASA traits of fresh and frozen-thawed semen, and all the sperm motility and velocity traits of fresh and frozen-thawed semen assessed by CASA were significantly interrelated among both the breeds. The interrelationships were stronger in Mehsana bulls as compared to Jafarabadi bulls.

162 FREEZING OF SEMEN FROM ENDANGERED ASTURIANA DE LA MONTAÑA BULLS IN ZWITTERONIC LIPIDS-BASED EXTENDERS

2008 ◽  
Vol 20 (1) ◽  
pp. 161 ◽  
Author(s):  
C. Tamargo Miguel ◽  
S. S. Pérez-Garnelo ◽  
P. Beltrán Breña ◽  
A. T. Palasz ◽  
J. De la Fuente ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Egg Yolk ◽  
Semen Parameters ◽  
Computer Assisted ◽  
Progressive Motility ◽  
Bull Semen ◽  
Straight Line ◽  
Before And After ◽  
Arcsine Transformation

This experiment was designed to test the efficacy of 2 different preparation protocols of zwitteronic soyabean-origin lipids for the production of lipidsglycerol liposomes for use in bull semen cryopreservation. Lipids liposomes were prepared at 10% concentration in Tris buffer by 1. highpressure homogenization (Panda 2K, Parma, Italy) and then 8% glycerol were added, extender-1 (E-1); Lipids were homogenized together with glycerol, extender-2 (E-2). Bioxcell extender (E-3) was used as control. Semen was collected 3 times from 3 endangered Asturiana de la Monta�a bulls by the means of an artificial vagina. Ejaculates with at least 70% motility were processed further by a standard freezing protocol used in our AI station. Semen was diluted at 37�C with each of the 4 extenders to a concentration of 92 � 106 spermatozoa per mL, cooled to 4�C over 4 h, aspirated into 0.25-mL plastic straws (IMV Technologies, Aigle, France), frozen in a bio-freezer (IMV Technologies) in 3 steps from 4 to –140�C, and then plunged into liquid N2. Straws were thawed in a water bath at 37�C for 30 s. Sperm motility was analyzed microscopically immediately after collection, after dilution, and after 4, 24, 48, and 72 h of storage at 4�C. Post-thaw semen progressive motility was assessed microscopically, and sperm movement characteristics were analyzed by computer-assisted semen analysis (CASA) (SCA�, Microptic, Barcelona, Spain). Data were compared between extenders and bulls by 2-way ANOVA; percentages were transformed by arcsine transformation before ANOVA. Total and progressive sperm motility at 0 h after dilution ranged from 90 to 70% and was not different between extenders and bulls. There was no difference between bulls in total and progressive motility after 24 h of cold storage; however, both were significantly greater (P < 0.05) for Control (62.4 � 14.7 and 41.4 � 14.9) and E-1 (70.1 � 12 and 33.8 � 7.0) extenders than for the E-2 extender (22.5 � 17 and 1.2 � 1.3). Average post-thaw sperm motility was not different between bulls for either extender, but motility for Bioxcell (Control, 48.1 � 14.6%) and E-1 extenders (43.2 � 13.0%) were significantly greater (P < 0.05) than for E-2 extender (18.7 � 8.8%). There were no differences between bulls for all kinematic semen parameters: curvilinear (VCL), straight line (VSL), average path (VAP) velocities, linearity (LIN) and straightness (STR), evaluated by CASA before and after freezing; however, all were lower (P < 0.05) for the E-2 extender and not different between Control and E-1 extenders. Sperm movements derived from heads (VCL) and linearity of sperm(LIN), both closely related to field fertility, were in the range of 90.9 � 2.1 and 63.0 � 5.5 for E-3 (Control) extender; 99.1 � 3.4 and 49.4 � 3.5 for E-1; and 21.8 � 2.2 and 29.9 � 4.0 for E-2. In summary, zwitteronic soyabean lipid liposomes are an effective egg yolk substitute for the cryopreservation of Asturiana de la Monta�a bull semen; however, the homogenization protocol of the lipids-glycerol mixture must be improved.

33 Caffeine improves equine sperm motility after thawing

2019 ◽  
Vol 31 (1) ◽  
pp. 142
Author(s):  
M. A. Lagares ◽  
N. C. Alves ◽  
A. L. A. Guimaraes ◽  
S. B. Luz ◽  
S. A. Diniz ◽  
...  
Keyword(s):  
Sperm Motility ◽  
The Other ◽  
Semen Sample ◽  
Computer Assisted ◽  
Sperm Transport ◽  
Head Displacement ◽  
Straight Line ◽  
Velocity Average ◽  
Lateral Head ◽  
Motility Parameters

The pattern of sperm transport and survival in the mare’s reproductive tract is different between fresh and frozen-thawed semen. A probable reason for this difference is the biophysiological changes in sperm during cryopreservation of equine semen. These changes can impair motility of stallion sperm after thawing. The aim of the present work was to test the effect of different caffeine concentrations on stallion sperm motility after thawing. One ejaculate of 9 stallions was frozen with the INRA82 frozen extender, and after thawing, different caffeine concentrations were added to the semen samples according to the treatments: control INRA82 without caffeine addition (T1), T1+1mM caffeine (T2), T1+2mM caffeine (T3), T1+3mM caffeine (T4), T1+5mM caffeine (T5), T1+7.5mM caffeine (T6), and T1+10mM caffeine (T7). The analysis of sperm motility parameters was performed with a computer-assisted semen analyser in 4 time periods: immediately after semen samples thawing (t0) and 15min (t15), 30min (t30), and 40min (t40) after semen sample thawing. One semen sample of each treatment was thawed, and an aliquot was analysed for the following computer-assisted semen analysis characteristics: velocity curvilinear (VCL; µm s−1), velocity straight line (µm s−1), velocity average path (µm s−1), linearity (%), straightness (%), wobble (%), amplitude of lateral head displacement (µm), beat cross frequency (BCF; Hz), and percentage of total sperm motility (TM) and progressive sperm motility. The statistical analysis was performed with ANOVA and Duncan’s test. The sperm parameters progressive sperm motility, linearity, wobble, and amplitude of lateral head displacement did not differ among the treatments (P&gt;0.05). Immediately after addition (t0) of 5, 7.5, and 10mM caffeine concentrations, an increase of TM was observed (T5: 53.1%; T6: 45.9%; and T7: 47.4%) compared with the other treatments (T1: 37.5%; T2: 36.0%; T3: 36.6%; and T4: 32.3%; P&lt;0.05). Although after 15min of incubation (t15) the TM decreased compared with t0 in T5, T6, and T7 treatments, the percentage was comparable with the other treatments at t15, t30, and t40. The mean value for TM was higher with 5mM caffeine compared with the control group (38.6% v. 34.7%; P&lt;0.05), whereas for the 10mM caffeine treatment velocity straight line (19.9v. 17.1µm s−1), velocity average path (25.6v. 22.9µm s−1), and straightness (75.4v. 72.3%) were higher than the control (P&lt;0.05). For the 5, 7.5, and 10mM caffeine treatments, VCL and BCF were higher than the control (VCL: 33.9, 34.5, 36.8, and 31.5µm s−1, respectively; BCF: 8.1, 8.6, 9.0, and 7.2Hz, respectively). The remaining motility parameters did not differ until 40min after the treatment (P&lt;0.05). In conclusion, the addition of 5, 7.5, and 10mM caffeine concentrations after semen thawing increased TM and most of the sperm motility characteristics. However, given the complexities of sperm transport, capacitation, and so on, further experiments are needed to test whether caffeine treatments could be used to improve the fertilization rate of frozen-thawed equine semen.

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.

scholarly journals Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1993
Author(s):  
Sabrina Gacem ◽  
Jaime Catalán ◽  
Anthony Valverde ◽  
Carles Soler ◽  
Jordi Miró
Keyword(s):  
High Resolution ◽  
Sperm Motility ◽  
Frame Rate ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Straight Line ◽  
Counting Chamber ◽  
Computer Assisted Sperm Analysis ◽  
Lateral Head

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.

scholarly journals Association between ambient particulate matter exposure and semen quality in fertile men

2022 ◽  
Vol 21 (1) ◽  
Author(s):  
Wei Wu ◽  
Yiqiu Chen ◽  
Yuting Cheng ◽  
Qiuqin Tang ◽  
Feng Pan ◽  
...  
Keyword(s):  
Particulate Matter ◽  
Sperm Motility ◽  
Semen Quality ◽  
Large Population ◽  
Sperm Concentration ◽  
Head Displacement ◽  
Straight Line ◽  
Distance Weighting ◽  
Sperm Development ◽  
Male Reproductive Health

Abstract Background Several studies have suggested adverse effects of particulate matter (PM) exposure on male reproductive health; few have investigated the association between PM exposure and semen quality in a large population of fertile men. Methods We evaluated 14 parameters of semen quality in 1554 fertile men in Nanjing from 2014 to 2016. Individual exposure to particular matter ≤10 μm in diameter (PM10) and ≤ 2.5 μm in diameter (PM2.5) during key periods of sperm development (0-90, 0-9, 10-14, 15-69, and 70-90 days before semen collection) were estimated by inverse distance weighting interpolation. Associations between PM exposure and semen quality were estimated using multivariable linear regression. Results Higher 90-days average PM2.5 was in association with decreased sperm motility (2.21% for total motility, 1.93% for progressive motility per 10 μg/m3 increase, P <  0.001) and four quantitative aspects of sperm motion (curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and amplitude of lateral head displacement (ALH), P <  0.01). The association between PM2.5 exposure and semen quality were generally stronger for the earlier exposure window (70-90 days prior to ejaculation) than for recent exposure (0-9, 10-14, or 15-69 days). In the subgroup of men who had normal sperm parameters (n = 1019), similar results were obtained. Ninety-days PM10 exposure was associated only with decreased VCL and VAP and was not related to sperm concentration. Conclusions Exposure to PM2.5 adversely affects semen quality, specifically lower sperm motility, in fertile men. Graphical abstract

14 FREEZING OF DONKEY SEMEN AFTER 24 HOURS OF COOL STORAGE: PRELIMINARY RESULTS

2013 ◽  
Vol 25 (1) ◽  
pp. 154 ◽  
Author(s):  
F. Qeusada ◽  
J. Dorado ◽  
D. Acha ◽  
I. Ortiz ◽  
M. Urbano ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Final Concentration ◽  
Computer Assisted ◽  
Preliminary Results ◽  
Cool Storage ◽  
Head Displacement ◽  
Straight Line ◽  
Negative Effect ◽  
Lateral Head ◽  
Motility Parameters

Several studies on sperm cooling and cryopreservation have been done in horses; however, only a few them have been developed in donkeys. In addition, no studies have been performed to freeze cooled stored donkey semen. Therefore, the aim of this study was to determine if it is possible to freeze donkey sperm after 24 h of cool storage. Semen was collected from 4 Andalusian donkeys by artificial vagina. After collection, each sample was separated into 2 aliquots; one of them was immediately frozen (t0) and the other one was cooled and stored before freezing (t24). The cryopreservation procedure consisted of a previous dilution of semen with EquiPro™. After that, semen was centrifuged and the sperm pellet resuspended with Gent® extender plus ethylene glycol (4%) to achieve a final concentration of 100 × 106 sperm mL–1. Sperm was slowly cooled to 5°C, loaded in 0.5-mL plastic straws and frozen in LN vapours. The second aliquot (t24) was diluted with Gent® extender to a final concentration of 50 × 106 sperm mL–1 and then cooled and stored at 5°C for 24 h. After that, cooled semen samples were cryopreserved following the same procedure as described above. Straws were thawed in a water bath at 37° for 30 s. Computer-assisted sperm motility analysis was performed. Total motility (TM), progressive motility (PM), and the following kinematic parameters: velocity curvilinear (VCL; µm s–1), velocity straight line (VSL; µm s–1), velocity average path (VAP; µm s–1), linearity (LIN; %), straightness (STR; %), wobble (WOB; %), amplitude of lateral head displacement (ALH; µm), and beat cross frequency (BCF; Hz) were compared between treatments by ANOVA. Results were expressed as mean ± standard error. Significant differences (P < 0.05) were found between treatments (t0 v. t24) for TM (63.76 ± 4.75 v. 51.67 ± 3.69), PM (36.01 ± 3.19 v. 27.24 ± 2.72), VCL (77.29 ± 0.65 v. 67.56 ± 0.78), VSL (58.50 ± 0.61 v. 52.11 ± 0.76), VAP (67.82 ± 0.64 v. 59.41 ± 0.79), LIN (57.90 ± 0.33 v. 59.53 ± 0.32), STR (70.39 ± 0.30 v. 72.43 ± 0.41), WOB (75.64 ± 0.22 v. 75.48 ± 0.32), ALH (1.88 ± 0.09 v. 1.69 ± 0.10), and BCF (6.28 ± 0.04 v. 6.51 ± 0.06). These preliminary results showed significant differences between cryopreservation at 0 and 24 h post-cooling; however, understanding that direct freezing is better in terms of sperm motility, cryopreservation of cooled stored semen could still be considered good according to the values obtained for sperm motility parameters after thawing. In our opinion, sperm centrifugation before cooling probably improve the results of cryopreservation 24 h post-cooling, due to the negative effect of seminal plasma on sperm viability during storage. In addition, the analysis of other sperm parameters would be useful to check more accurately differences between treatments. In conclusion, sperm motility parameters were higher in donkey semen samples immediately frozen after collection in comparison to semen samples cryopreserved after 24 h of cooling storage. Further studies are needed to improve cooling and cryopreservation procedures for freezing cooled stored donkey semen.

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