Morphological Image of Fresh and Cryopreserved Dog Semen Evaluated by the Strict Analysis of Sperm Morphology Method, Using Sperm Quality Analyzer (SQA IIc) Evaluation

Thirty fresh ejaculates from 15 dogs were cryopreserved in Tris-fructose-citric acid-egg-yolk extender with a glycerol content of 6%. Semen samples were examined by the methods of routine sperm analysis and by the SQA IIc device. The routine semen examination focused on the evaluation of parameters determining the quality of sperm membranes. The significance of monitoring semen quality in the course of the short-term survival test for predicting dog semen quality after thawing was assessed. Relevance of the assessment of sperm morphology, and above all the percentage of sperm with membrane changes in the acrosomal region was documented. The fact that the SQA device analyses semen quality by evaluating the mass of moving cells was confirmed. The results provided by the SQA IIc device appear insufficient for the needs of deeper dog semen analysis, especially morphology assessment.

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Validation and Usefulness of the Sperm Quality Analyzer (SQA II-C) for Bull Semen Analysis

Reproduction in Domestic Animals ◽

10.1111/j.1439-0531.2005.00584.x ◽

2005 ◽

Vol 40 (3) ◽

pp. 237-244 ◽

Cited By ~ 10

Author(s):

G Hoflack ◽

T Rijsselaere ◽

D Maes ◽

J Dewulf ◽

G Opsomer ◽

...

Keyword(s):

Sperm Quality ◽

Semen Analysis ◽

Bull Semen ◽

Quality Analyzer ◽

Sperm Quality Analyzer

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Assessment of Sperm Quality - A Light Microscope Study

Journal of Evolution of Medical and Dental Sciences ◽

10.14260/jemds/2021/299 ◽

2021 ◽

Vol 10 (19) ◽

pp. 1417-1421

Author(s):

Jyothi A. Raj ◽

Heera Sankar ◽

Sagarika Mahapatra ◽

Ashima Binny

Keyword(s):

Light Microscope ◽

Sperm Morphology ◽

Clinical Laboratory ◽

Sperm Quality ◽

Semen Analysis ◽

Complete Evaluation ◽

Normal Sperm ◽

Middle Piece ◽

Sperm Counts ◽

Work Up

BACKGROUND Semen analysis is an integral part of work up for infertility in men, with sperm morphology being an important qualitative parameter. Qualitative defects can affect any part of the sperm and are classified as defects in the head, middle piece, and tail, based on morphology. The focus of the study was to assess qualitative defects in sperms by light microscopy, in semen with normal sperm counts. METHODS This study is hospital based, descriptive, retrospective study. Of the semen samples received in the clinical laboratory, fifty with normal sperm counts were included in the study and processed according to standard protocol. For evaluation of qualitative defects by sperm morphology, smears were fixed in ethanol, stained with Papanicolaou stain [PAP], and assessed under light microscope. RESULTS The 50 semen samples included in the study had sperm counts ranging from 15 to 80 million / ml. Thirty samples had less than 10 % abnormal forms, fourteen samples had 11 - 20 % abnormal forms, five samples had 21 - 30 % abnormal forms and one sample had 40 % abnormal sperms. Qualitative defects were classified as morphological abnormalities in head, neck, and tail. Of the fifty cases, most defects were found in the head, followed by those in the neck and tail. Common defects noted were double head (44 %), abnormal sized heads, and bent neck (48 %). Coiling was a common defect noted in the tail (10 %). Most sperms showed a combination of defects. CONCLUSIONS Qualitative defects in sperm morphology are often seen in samples with normal sperm counts. Assessment of microscopic characteristics of human spermatozoa is as important as count and motility in the complete evaluation and work-up of semen samples in cases of infertility. KEY WORDS Semen, Sperm, Quality, Microscopy, Morphology

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Validation and usefulness of the Sperm Quality Analyzer V equine for equine semen analysis

Theriogenology ◽

10.1016/j.theriogenology.2010.08.003 ◽

2011 ◽

Vol 75 (1) ◽

pp. 189-194 ◽

Cited By ~ 6

Author(s):

M. Hoogewijs ◽

S. De Vliegher ◽

C. De Schauwer ◽

J. Govaere ◽

K. Smits ◽

...

Keyword(s):

Sperm Quality ◽

Semen Analysis ◽

Quality Analyzer ◽

Sperm Quality Analyzer

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

Zygote ◽

10.1017/s0967199418000230 ◽

2018 ◽

Vol 26 (4) ◽

pp. 301-307 ◽

Cited By ~ 2

Author(s):

José A. B. Bezerra ◽

Andréia M. Silva ◽

Patrícia C Sousa ◽

Lívia B. Campos ◽

Érica C. G. Praxedes ◽

...

Keyword(s):

Sperm Quality ◽

Semen Analysis ◽

Membrane Integrity ◽

Egg Yolk ◽

Coconut Water ◽

Computer Assisted ◽

Epididymal Sperm ◽

Sperm Plasma Membrane ◽

Pecari Tajacu ◽

Functional Membrane

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

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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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Effect of whole and clarified egg yolk based extenders on post-thaw semen quality in bulls.

10.31220/agrirxiv.2021.00050 ◽

2021 ◽

Author(s):

Prosper Kamusasa ◽

Eddington Gororo ◽

Fungayi Primrose Chatiza

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

Egg Yolk ◽

Quality Parameters ◽

Normal Morphology ◽

Semen Cryopreservation ◽

Bull Semen ◽

Inclusion Level ◽

Whole Egg

Abstract This study was conducted to evaluate the comparative cryoprotective effects of whole egg yolk and clarified egg yolk on post thaw sperm quality parameters and to determine the optimum clarified egg yolk inclusion level (10-20%) in semen extenders for Mashona bull semen cryopreservation. It was shown that there was a significant decrease in sperm quality variables following cryopreservation. Semen quality increased with the concentration of clarified egg yolk, indicating a positive relationship between egg yolk LDL concentration and maintenance of in vitro sperm quality. The 20% clarified egg yolk (CEY20) extender treatment gave post-thaw motility, viability and normal morphology values which were comparable to the control (20% whole egg yolk, WEY20). The 10% clarified egg yolk concentration gave the least post-thaw quality values and the greatest proportion of defective spermatozoa. This experiment found no advantage of replacing whole egg yolk with up to 15% clarified egg yolk in Mashona bull semen cryopreservation. However, 20% clarified and 20% whole egg yolk performed similarly in the maintenance of post-thaw sperm motility, viability and normal morphology.

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Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo

Animals ◽

10.3390/ani9060304 ◽

2019 ◽

Vol 9 (6) ◽

pp. 304 ◽

Cited By ~ 1

Author(s):

Giusy Rusco ◽

Michele Di Iorio ◽

Pier Paolo Gibertoni ◽

Stefano Esposito ◽

Maurizio Penserini ◽

...

Keyword(s):

Brown Trout ◽

Semen Quality ◽

Sperm Quality ◽

Egg Yolk ◽

Dna Integrity ◽

Experimental Conditions ◽

C 30 ◽

Thawing Rate

The aim of our study was to test the effects of different non-permeating cryoprotectants (NP-CPAs), namely low-density lipoproteins (LDLs), sucrose, and egg yolk, and thawing rates on the post-thaw semen quality and fertilizing ability of the native Mediterranean brown trout. Pooled semen samples were diluted 1:3 (v:v) with 2.5%, 5%, 10%, or 15% LDL; 0.05, 0.1, or 0.3 M sucrose; or 10% egg yolk. At the moment of analysis, semen was thawed at 30 °C/10 s or 10 °C/30 s. The post-thaw semen quality was evaluated, considering motility, the duration of motility, viability, and DNA integrity. Significantly higher values of motility and viability were obtained using egg yolk/10 °C for 30 s, across all treatments. However, LDL and sucrose concentrations affected sperm cryosurvival, showing the highest post-thaw sperm quality at 5% LDL and 0.1 M sucrose. Based on the in vitro data, egg yolk, 5% LDL, and 0.1 M sucrose thawed at 10 °C or 30 °C were tested for the in vivo trial. The highest fertilization and hatching rates were recorded using egg yolk/10 °C (p < 0.05). According to these in vitro and in vivo results, egg yolk emerged as the most suitable NP-CPA and 10 °C/30 s as the best thawing rate for the cryopreservation of this trout sperm, under our experimental conditions.

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Influence of Lifestyle and Environmental Factors on Semen Quality in Ghanaian Men

International Journal of Reproductive Medicine ◽

10.1155/2020/6908458 ◽

2020 ◽

Vol 2020 ◽

pp. 1-7

Author(s):

Richard Michael Blay ◽

Abigail Duah Pinamang ◽

Augustine E. Sagoe ◽

Ewurama Dedea Ampadu Owusu ◽

Nii Koney-Kwaku Koney ◽

...

Keyword(s):

Environmental Factors ◽

Male Infertility ◽

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Sperm Count ◽

Semen Analysis ◽

Reproductive Age ◽

Adult Males ◽

Lifestyle Habits

Introduction. Male infertility is known to contribute about half of all infertility cases. In Ghana, the prevalence of male infertility is higher (15.8%) than in females (11.8%). Sperm quality is associated with the likelihood of pregnancy and known to be the cause of male fertility problems 90% of the time. Exposure to certain environmental factors reduces semen quality in men. The study examined the effects of environmental and lifestyle factors on semen quality in Ghanaian men. Materials and Methods. This was a cross-sectional study involving 80 apparent healthy adult males in their reproductive age. Participants were males referred to the laboratory (Immunology Unit of the Korle-Bu Teaching Hospital) for semen analysis test and/or culture and sensitivity. Participants were made to fill out a questionnaire which entailed selected environmental factors (accidents or trauma, exposure to chemicals, radiation, and heat) and lifestyle habits (including alcohol consumption, smoking, and whether participants sat more or less than 4 hours per day). Semen samples were then collected by masturbation into sterile containers and analysed in accordance with WHO guidance for semen analysis within 60 minutes after ejaculation and collection. Results. About 69% of participants had semen pH within the normal range compared to 15% whose pH were lower than 7.2. There was a significantly high number of immotile sperm cells ( p value = 0.017) in participants who sat for more than 4 hours as compared to those that sat for less than 4 hours in a day. Active sperm motility and viability showed significant increase ( p value = 0.002 and 0.009, respectively) in participants who kept their cell phones in their side pockets. Smoking produced a twofold decrease in sperm count as smokers had a significantly lower sperm count ( 12.28 ± 10.95 × 10 6 /ml) compared to the smoke-free ( 23.85 ± 22.14 × 10 6 /ml). For exposure to STDs, no significant differences were recorded among study groups concerning semen quality. Conclusion. Sperm quality in Ghanaian men is associated with lifestyle habits. Smoking and sitting for long hours influenced sperm motility and count, respectively. Knowledge of the factors that influence sperm quality in this geographical region can contribute to informed decisions on effective management of infertility in Ghanaian men.

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Effect of temperature and time after collection on buck sperm quality

BMC Veterinary Research ◽

10.1186/s12917-019-2135-y ◽

2019 ◽

Vol 15 (1) ◽

Author(s):

Kirsten Hahn ◽

Klaus Failing ◽

Axel Wehrend

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

Semen Analysis ◽

Effect Of Temperature ◽

Sperm Viability ◽

Factors Affecting ◽

Semen Collection ◽

Examination Time ◽

Collection Period ◽

Time Point

Abstract Background Different parameters are assessed as part of the semen analysis but a standard protocol for evaluation of goat semen is still missing. The aim of this study was to analyse two different factors affecting buck sperm quality in the post-collection period prior to adding the extender. Here we examined the effects of two handling temperatures (20 °C, 37 °C) and various examination time points (3–30 min) after semen collection. Results Examination time point had a significant influence on raw sperm viability (p < 0.05), motility (p < 0.05) and on semen pH (p < 0.05). The two different handling temperatures had no significant effect on sperm viability (p > 0.05), motility (p > 0.05), with the exception of fast moving sperm (p = 0.04), or on semen pH (p > 0.05). Conclusion Examination time point was identified as factor strongly influencing raw peacock buck semen after collection. Raw goat semen can tolerate room temperatures for at least 10 min without impacting overall semen quality. In order to obtain comparable results, semen samples should always be examined within 10 min after collection.

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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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