Fresh semen characteristics in captive accipitrid and falconid birds of prey

Artificial insemination (AI) is the most frequently used assisted reproductive technique for captive propagation of rare avian species. As semen quality is critical for reproductive success, baseline data are needed for evaluating and selecting the best male bird donors. To this end, we used computer-assisted semen analysis to assess male eastern imperial eagles (n = 7), northern goshawks (n = 24) and peregrine falcons (n = 20). While imperial eagles and northern goshawks donate ejaculate voluntarily, peregrine falcons required cloacal massage. Eight peregrine falcon females were inseminated with semen from eight males, with fresh ejaculates (15 to 50 µl) applied to the pars uterina of the oviduct immediately after collection and examination. All females were inseminated within 2 h of laying an egg. A fertilization rate of 70% was achieved using this method. Minimum semen characteristics associated with egg fertilization included a semen concentration of 115.12 × 106/ml, 33.52% total motility, 1.92% spermatozoa with progressive motility and 0.17% with rapid motility. Comparative data on spermatozoa concentration and kinematics suggest that eastern imperial eagles concentrate on high quality semen investment at the start of the breeding season, northern goshawks compensate for a decrease in motility-associated parameters with increased semen concentration and peregrine falcons maintain semen production standards throughout the breeding season. Our data show that, in birds of prey, levels of egg fertilization following AI with fresh semen can be almost as successful as after natural mating.

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

Archives Animal Breeding ◽

10.5194/aab-63-145-2020 ◽

2020 ◽

Vol 63 (1) ◽

pp. 145-154 ◽

Cited By ~ 1

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.

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105 Increasing fertility of interspecific hybrid males using biotechnological approaches

Reproduction Fertility and Development ◽

10.1071/rdv33n2ab105 ◽

2021 ◽

Vol 33 (2) ◽

pp. 159

Author(s):

A. Vetokh ◽

A. Tadzhieva ◽

B. Iolchiev ◽

N. Volkova ◽

V. Bagirov

Keyword(s):

Dna Fragmentation ◽

Interspecific Hybrid ◽

Semen Quality ◽

Sperm Quality ◽

Semen Analysis ◽

Differential Staining ◽

Computer Assisted ◽

Sperm Dna Fragmentation ◽

Russian Science

The results of AI depend on many factors, with the quality of semen being one of the most important. Not all male hybrids can meet the requirements for semen quality, because they often have reduced fertility following cryopreservation. Thus, it is necessary to improve semen processing before use in AI. The aim of the study was to evaluate the effectiveness of using the “swim-up” flotation method to improve sperm quality of hybrid males of the Ovis genus. Semen from interspecific hybrid rams (1/4 Argali×3/4 Romanov, n=15; 1/8 Argali×7/8 Romanov, n=15) was freshly obtained, frozen–thawed, and processed by the swim-up method. Evaluation of sperm motility was determined using computer-assisted semen analysis. Statistical analysis was performed using SPSS vs.15.0 (ANOVA and t-test; SPSS Inc.). Semen was collected during the breeding season (October–December) via artificial vagina. Assessment of acrosome integrity was determined using differential staining with a Diachem diff-quick kit (NPF ABRIS+). The degree of sperm DNA fragmentation was determined using the acridine-orange test. The sperm freezing/thawing cycle was accompanied by sperm damage and an increase in the proportion of immobile sperm from 10 to 58%, with non-progressive movement increasing from 9 to 19.3%. The number of spermatozoa with abnormal morphology doubled, and the DNA fragmentation index increased from 16 to 26%. Use of the swim-up procedure allowed us to sort progressively motile spermatozoa. The content of progressively motile spermatozoa in the samples obtained from the supernatant was 86%, which was 2.3 times higher than in frozen–thawed sperm (P≤0,01). The obtained results show the effective use of the swim-up procedure to determine the quality of semen in hybrid rams. These studies were carried out with financial support from the Russian Science Foundation, grant No. 18-16-00079 and the Ministry of Science and Higher Education of the Russian Federation.

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69 Invitro production of ovine embryos using either fresh or frozen-thawed semen

Reproduction Fertility and Development ◽

10.1071/rdv33n2ab69 ◽

2021 ◽

Vol 33 (2) ◽

pp. 141

Author(s):

G. Márquez-Márquez ◽

A. Velázquez-Roque ◽

F. Villaseñor-González ◽

M. Kjelland ◽

H. Álvarez-Gallardo ◽

...

Keyword(s):

Semen Quality ◽

Small Ruminants ◽

Physiological Saline ◽

Ovis Aries ◽

Penicillin G ◽

Domestic Sheep ◽

Assisted Reproductive Technique ◽

Physiological Saline Solution ◽

Α Level ◽

Fresh Semen

Invitro embryo production (IVP) is an important tool for genetic improvement in small ruminants. Semen quality is one of the most important aspects to consider for the success of this assisted reproductive technique. With ovine IVP, it is a common practice to use fresh semen for IVF, but this could be a problem because the differences between ejaculates from the same animal are well documented and a source of variation in IVP results. The objective of this research was to compare the effect of fresh and frozen–thawed domestic sheep (Ovis aries) semen on IVF for ovine IVP. The research was carried out in the reproduction laboratory at the Palominos Ranch (Jalisco, México). The IVP was performed with a continuous invitro culture system. Ovaries (n=186) were collected from a slaughterhouse (León, México) and transported to the laboratory within 2h in physiological saline solution (0.9% NaCl) supplemented with penicillin G (100IU mL−1) and streptomycin sulphate (100µg mL−1). For IVP, IVF-Bioscience™ media were used for IVM, IVF, and invitro culture (IVC). For IVM, the cumulus–oocyte complexes (COCs) were selected (only grades 1 and 2) and matured for 24h at 38.5°C in 5% CO2 in air and 100% humidity. Matured oocytes (n=1000) were invitro fertilized using either fresh or frozen–thawed semen (Triladyl™; Minitube) from the same sheep, at a concentration of 2×106 sperm mL−1, for 18h in 38.5°C, 5% CO2 in air, and 100% humidity. The presumptive zygotes were denuded by pipetting and set in IVC until Day 7 at 38.5°C, 5% CO2, 5% O2, and 90% N2 at 100% humidity. The percentages of cleavage, embryos with more than 6 cells, and blastocysts on Day 7 of culture were evaluated, based on the initial number of oocytes entering into IVM. Statistical analyses were carried out with the GLM procedure of SAS software (version 9.3; SAS Institute Inc.) to evaluate the results of fresh versus frozen–thawed (α level=0.05). Rates of cleavage, embryos with more than 6 cells, and blastocysts on Day 7 were similar (P>0.05): fresh 52.3±3.0%, 43.6±2.6%, and 34.3±2.9%, respectively; frozen–thawed: 53.3%±3.0, 41.1±2.6%, and 33.3±2.9%, respectively. In conclusion, under the conditions of this research, the use of fresh and frozen–thawed semen had similar results for ovine IVP.

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A Review on Animals Semen Characteristics: Fertility, Reproduction and Development

Asian Journal of Advances in Agricultural Research ◽

10.9734/ajaar/2019/v10i230024 ◽

2019 ◽

pp. 1-9

Author(s):

Farzad Moradpour

Keyword(s):

Structural Integrity ◽

Current Knowledge ◽

Semen Analysis ◽

Kinetic Property ◽

Genetic Material ◽

Computer Assisted ◽

Semen Characteristics ◽

The Individual

In this research, the goal of review was summarizing the current knowledge of the methods available to assess in vitro quality of frozen-thawed bovine spermatozoa also, a review on animal’s semen characteristics: fertility, reproduction and development after AI with that semen. Artificial insemination (AI) is the first generation reproductive biotechnology that has made a deep contribution to the genetics improvement in several animals. A fertile ejaculate must meet certain semen characteristics quality standards, such as: normal morphology, active energy metabolism, progressive motility, structural integrity and functionality of the membrane, penetration capacity and optimum transfer of genetic material. The percentage of total motile spermatozoa in normal canine ejaculates is between 70 to 90%. By the way, there are a lot of parameters that able to change on the composition and structure of various sperm plasma member domains, such as change temperature and sensitive to any theirs environments in vivo and vitro (tropical climates), season also nutrition. Computer-assisted semen analysis (CASA) is primarily used to obtain accurate and objective kinetic sperm measurements that gives extensive information about the kinetic property of the ejaculate based on measurements of the individual sperm cells.

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18 SEMENRATE: THE USE OF COMPUTER-ASSISTED SEMEN ANALYSIS AND FLOW CYTOMETRY FOR OBJECTIVE BOVINE SEMEN ANALYSIS IN THE UNITED KINGDOM

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab18 ◽

2017 ◽

Vol 29 (1) ◽

pp. 116

Author(s):

M. W. Spilman ◽

K. L. Burton ◽

J. M. E. Statham

Keyword(s):

Flow Cytometry ◽

United Kingdom ◽

Semen Quality ◽

Semen Analysis ◽

Computer Assisted ◽

Motile Sperm ◽

Bovine Semen ◽

The United Kingdom ◽

Frozen Semen ◽

Viable Sperm

Routine assessment of bovine semen consists of a subjective assessment of morphology, motility and concentration. This subjective approach used during quality control at semen production centres (SPC) or investigations of poor reproductive performance in veterinary practice has been shown to be relatively inaccurate, imprecise, and operator dependent (Vincent, et al. 2012 Anim. Reprod. 9, 153–165). Assessment of frozen semen samples in a dedicated laboratory aimed to establish variations in multiple parameters associated with fertility using computer-assisted semen analysis and flow cytometry and evaluate their relationship to semen performance in the field. This has developed into a commercial service that is available to veterinarians and farmers across the United Kingdom. AI semen from 50 farms across Yorkshire, UK, that had been stored on farm was assessed for factors associated with fertility (motility, progressive motility, intact acrosome, viability, and polarised mitochondria). Data ranges and mean values for each parameter have been analysed. This analysis is ongoing as the dataset continues to expand and significance will be assessed. For frozen semen (n = 79), % viable sperm (max = 67.64, min = 0.00, mean = 43.44), % sperm with polarised mitochondria (max = 72.50, min = 0.26, mean = 38.56), % sperm with acrosome intact (max = 68.82, min = 0.06, mean = 35.29), % motile sperm (max = 66.90, min = 0.00, mean = 37.44) and % progressively motile sperm (max = 59.00, min = 0.00, mean = 26.11). 25% of the samples fell below the cut off for release of 30% motile sperm set by SPCs. For sexed AI semen (n = 9), % viable sperm (max = 66.31, min = 17.08, mean = 43.57), % polarised mitochondria (max = 26.74, min = 13.40, mean = 19.96), % intact acrosome (max = 52.62, min = 15.34, mean = 37.00), % motile (max = 38.00, min = 9.40, mean = 24.88) and % progressively motile (max = 22.80, min = 3.90, mean = 13.15). Objective semen analysis before beginning an embryo collection programme allows informed decisions to be made regarding semen choice and dosage depending on compensable v. non-compensable defects detected (Hudson et al. 2012 Dairy Herd Health 73–111; CABI Publishing). Use of semen that falls below the 30% cut off for SPCs is unlikely to perform as expected in the field (Phillips et al. 2004 Anim. Reprod. Sci. 80, 47–61). A European collaboration aims to establish correlations between semen quality parameters and fertility outcomes for UK cattle herds, providing unique data for the industry (Sellem et al. 2015 Theriogenology 84, 1447–1454.e5). These data should highlight to stakeholders in the industry how imperative optimal semen quality is and highlight the benefits to herd fertility and financial performance.

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

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab180 ◽

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.

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116. EFFECTS OF SUPPLEMENTAL VITAMIN ON THE SEMEN CHARACTERISTICS OF MARKHOZ GOATS DURING NON-BREEDING SEASON

Reproduction Fertility and Development ◽

10.1071/srb09abs116 ◽

2009 ◽

Vol 21 (9) ◽

pp. 35

Author(s):

A. Towhidi ◽

A. Farshad ◽

M. Dolatpanah ◽

R. Salehi

Keyword(s):

Vitamin E ◽

Breeding Season ◽

Semen Quality ◽

Sperm Concentration ◽

Progressive Motility ◽

Semen Volume ◽

Semen Characteristics ◽

Supplemental Vitamin ◽

Diet Control ◽

Live Sperm

The aim of the study was to investigate the effect of dietary supplementation of vitamin E on the semen characteristics of the Markhoz goats. Eighth bucks were randomly allocated into two groups, and received two different diets: unsupplemented diet (control) and supplemented diet with vitamin E (0.30 g/Kg DM). All experimental diets were formulated according to AFRC 1998. Semen was collected at 14-days intervals from June 17, 2006 to September 2, 2006 (non-breeding season) using artificial vagina. Semen characteristics were evaluated including semen volume, sperm concentration, live sperm percentage, percentage of motility and progressive motility. This characteristics were evaluated at two weeks intervals the trial. The total number of spermatozoa per ejaculation was calculated by multiplication of the semen volume with sperm concentration. Sperm motility was also analyzed by placing a sample on pre-warmed (37 º C) microscope slide covered with a cover slip, and examined under a high power microscope at a magnification × 200. Data was analyzed using proc MIXED in SAS program. Significant effect (P≤0.05) of the week (sampling time) was observed for all the parameters except for semen volume. Vitamin E supplementation significantly improved (P≤0.05) total number (Control: 263.7 ×107±17.506 vs. Vit E: 320.95 ×107±17.506) and sperm concentration (Control: 301.79 ×107±13.657 vs. Vit E: 386.57 ×107± 13.657), motility (Control: 77.27% ±0.89 vs. Vit E: 82.6% ±0.89) and progressive motility (Control: 4.208 ±0.138 vs. Vit E: 4.229 ±0.138), percentage of viability (Control: 80.57% ±0.89 vs. Vit E: 85.9% ±0.89). The results suggested that the supplemental Vitamin E may improve the semen quality and fertility in the Markhoz goats.

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P–011 Automated sperm morphology assessment using artificial intelligence technology

Human Reproduction ◽

10.1093/humrep/deab130.010 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

A Agarwal ◽

M K Panne. Selvam

Keyword(s):

Artificial Intelligence ◽

Predictive Value ◽

Sperm Morphology ◽

Semen Quality ◽

Semen Analysis ◽

Sperm Concentration ◽

Ease Of Use ◽

Computer Assisted ◽

Diagnostic Use ◽

Normal Sperm

Abstract Study question Can LensHooke X1 PRO semen analyzer be used to evaluate sperm morphology in men with infertility? Summary answer Morphology results generated by X1 PRO are highly reliable when normal sperm forms are ≥4% and therefore they can be reported in such cases . What is known already Most laboratories rely on manual evaluation of sperm morphology smears, which is a time-consuming procedure and its results are subjected to a relatively high variability. However, in recent years the computer-assisted semen analyzers are being increasingly used to evaluate sperm morphology. The X1 PRO semen quality analyzer was designed for in vitro diagnostic use to analyze sperm concentration, total, progressive and non-progressive motility as well as sperm morphology based on WHO 5th edition criteria. Evaluation of sperm morphology using X1 PRO based on AIOM (Artificial Intelligence Optical Microscopic)-based technology requires no fixation steps or staining unlike the manual method. Study design, size, duration This cross-sectional study used 31 semen samples from 8 normozoospermic healthy volunteers and 5 infertile men with a minimum abstinence period between 2 - 3 days. While the 8 healthy semen donors produced a total of 26 ejaculates, which were split into 88 aliquots, the 5 infertile patients produced 5 ejaculates that were split into 13 aliquots. Participants/materials, setting, methods A total of 101 aliquots were prepared from the native semen samples either by dilution or concentration using seminal plasma of the respective donors. Automated semen analysis was performed by the X1 PRO semen analyzer and the results of sperm morphology were compared with manual morphology results using Diff-Quik staining. Statistical analysis was carried out to calculate the positive predictive value (PPV) and negative predictive value (NPV) of X1 PRO semen analyzer. Main results and the role of chance The X1 PRO sperm morphology results show a weak non-significant (P = 0.2441) correlation (r = 0.119) with the manual results. However, X1 PRO demonstrated a high PPV (97.7%) and a low NPV (9.1%) for correct assessment of sperm morphology (≥4%) when compared to manual results. Due to its high PPV, laboratories can report the morphology results generated by X1 PRO in all such cases when normal sperm forms are ≥4%. However, a manual evaluation is necessary in patients with abnormal morphology (<4%). Limitations, reasons for caution One of the limitation of this study is that X1 PRO morphology values did not correlate with manual results. The low NPV seen in our study is due to the inclusion of very few samples with abnormal sperm forms (<4%) in the analysis. Wider implications of the findings: The X1 PRO’s combination of speed, ease of use, accuracy and portability makes it a good choice of device for small medical offices to large IVF centers. High PPV of X1 PRO allows it to correctly identify normal sperm forms for diagnostic use. Trial registration number 18–771

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