Using videotaped specimens to test quality control in a computer-assisted semen analysis system

The objective of the study was a comparison of two protocols of goat semen cryopreservation with the use of computer-assisted semen analysis system. Twenty ejaculates obtained with electroejaculation method were assessed. Each ejaculate was divided in half and frozen according to two protocols. In protocol I semen was centrifuged in order to remove its plasma and diluted in Tris buffer extender containing glucose, citric acid and glycerol with 20% addition of egg yolk. Protocol II did not include removal of plasma and the extender contained 1.5% egg yolk. It was shown that the removal of semen plasma improved motility of goat spermatozoa following freezing/thawing with respect to the following motility indicators: motility, average path velocity, amplitude of lateral head displacement at p < 0.05, and straight velocity, straightness and linearity at p < 0.01. In conclusion, the removal of semen plasma through centrifugation improved motility properties of goat semen following the freezing/thawing procedure.

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57 EFFECT OF CHOLESTANOL OR CHOLESTEROL ON THE MOTILITY OF FROZEN GOAT SPERMATOZOA AFTER A THERMAL RESISTANCE TEST

Reproduction Fertility and Development ◽

10.1071/rdv26n1ab57 ◽

2014 ◽

Vol 26 (1) ◽

pp. 142

Author(s):

B. G. Silva ◽

E. A. Moraes ◽

W. C. G. Matos ◽

C. S. Oliveira ◽

W. D. Ferrari Junior ◽

...

Keyword(s):

Thermal Resistance ◽

Liquid Nitrogen ◽

Semen Analysis ◽

Egg Yolk ◽

Water Bath ◽

Resistance Test ◽

Computer Assisted ◽

Progressive Motility ◽

Working Solution ◽

Analysis System

The objective of the present study was to determine the concentration of cholesterol or cholestanol-loaded-cyclodextrin that needs to be added to goat sperm before cryopreservation to optimize its survival. The cholesterol or cholestanol loaded methyl-β-cyclodextrin was prepared as described by Moraes et al. (2010 Anim. Reprod. Sci. 118, 148–154). A working solution of the cholesterol or cholestanol-loaded cyclodextrin was prepared by adding 50 mg of each one to 1 mL of TALP at 37°C and mixing the solution briefly using a vortex mixer. Ejaculates (n = 24) from 5 bucks were used for this experiment. Sperm from each ejaculate were diluted 1 : 1 (vol : vol) in Tris diluent (200 mM Tris, 65 mM citric acid, and 55 mM glucose) and centrifuged at 800 × g for 10 min. The pellets were resuspended to a concentration of 120 × 106 sperm mL–1 in Tris and subdivided into 7 aliquots of 5 mL each (600 × 106 total sperm). Sperm were treated in 7 treatment groups that received no additive (0 mg; control) or different levels of cholesterol or cholestanol (0.75, 1.5, or 3.0 mg/120 × 106 sperm). All treatments were incubated for 15 min at room temperature and then cooled to 4°C over 2 h. The samples were diluted with Tris-egg yolk diluent containing 2% glycerol. The sperm were packaged into 0.5-cc straws and frozen in static liquid nitrogen vapor for 20 min and then straws were plunged into liquid nitrogen and stored until analysed for motility and thermal resistance test using a computer-assisted semen analysis system (CASA). Two straws from each treatment were thawed in a 37°C water bath for 30 s and extended in Tris. For the thermal resistance test, after thawing, 0.5 mL of semen from each treatment was placed in 1.5-mL tubes in a water bath at 37°C for 3 h. At 0, 60, 120, and 180 min, subsamples were evaluated for sperm total and progressive motility using a computer-assisted sperm motion analyzer. A total of 200 spermatozoa were counted in at least 5 different fields. Data were analysed using ANOVA and treatment means were separated, using the SNK test at 5% probability. Cholesterol (0.75 mg; 46.7%) and cholestanol (1.5 mg; 40.5%) produced an increase in progressive motility compared with other treatments after 1 h of incubation (P < 0.05). However, cholestanol (0.75 mg; 39.5 and 31%) was higher for total and progressive motility after 3 h of sperm incubation compared with the control (27 and 17.8%; P < 0.05), respectively. The addition of 0.75 mg of cholestanol in fresh sperm before cryopreservation improved the motility of freeze-thawed goat sperm compared with cholesterol. Therefore, adding cholestanol to goat sperm membranes improved cell cryosurvival. Supported by Fundação de Amparo à Ciência e Tecnologia de Pernambuco (FACEPE) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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Validation of a smartphone-based, computer-assisted sperm analysis system compared with laboratory-based manual microscopic semen analysis and computer-assisted semen analysis

Investigative and Clinical Urology ◽

10.4111/icu.2019.60.5.380 ◽

2019 ◽

Vol 60 (5) ◽

pp. 380 ◽

Cited By ~ 3

Author(s):

Won Hee Cheon ◽

Hyun Jun Park ◽

Min Jung Park ◽

Mi Young Lim ◽

Jih Hoon Park ◽

...

Keyword(s):

Semen Analysis ◽

Computer Assisted ◽

Sperm Analysis ◽

Computer Assisted Sperm Analysis ◽

Analysis System

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Optimization of a continuous real-time computerized semen analysis system for ram sperm motility assessment, and evaluation of four methods of semen preparation

Reproduction Fertility and Development ◽

10.1071/rd9960219 ◽

1996 ◽

Vol 8 (2) ◽

pp. 219 ◽

Cited By ~ 21

Author(s):

WV Holt ◽

MJ Palomo

Keyword(s):

Real Time ◽

Semen Analysis ◽

Computer Assisted ◽

Assessment And Evaluation ◽

Analysis System ◽

Set Up ◽

Assisted Analysis ◽

Ram Sperm ◽

Preparation Techniques

Sampling conditions that affect the biological validity of computer-assisted analysis of ram sperm motion were examined using a continuous real-time computerized semen analysis system (Hobson Sperm Tracker). Search radius (SR, 10 settings) and minimum track point (MTP, 10 settings) were varied factorially to evaluate their effects on the inclusion of sperm subpopulations within derived datasets. Low SR (< 12 microns) or high MTP values (> 26 frames) precluded measurements of rapidly moving cells, whereas high SR (> 17 microns) and low MTP settings (< 22 frames) led to erroneous tracking and poor data quality. Suitable settings for these set-up parameters were derived and tested for their biological consistency with four methods of preparing ram semen for computerized analysis. The preparation techniques tested were: centrifugation through sucrose-based Ficoll and Percoll media, a swim-up technique and simple dilution in Tyrode's media. The 'selective' Percoll and swim-up methods generated sperm populations with significantly higher linear velocities and a lower tendency to deviate from linear trajectories than from either the Ficoll method or dilution technique. Deleterious effects of centrifugation were evident, particularly on sperm survival in vitro over several hours. It is concluded that computer-assisted semen analysis provides useful information about the behaviour of ram spermatozoa in vitro, but the measurement conditions must be defined carefully.

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Влияние биологической полноценности спермы быков-производителей канадской селекции на репродуктивные показатели коров отдельного стада

Аграрная Россия ◽

10.30906/1999-5636-2017-2-18-21 ◽

2017 ◽

pp. 18-21

Author(s):

Александр Александрович Некрасов ◽

Николай Александрович Попов ◽

Байлар Садраддинович Иолчиев

Keyword(s):

Semen Analysis ◽

Computer Assisted

Исследовано влияние качества спермы быков-производителей на репродуктивные показатели стада. Материалом для исследования послужила замороженно-оттаянная сперма быков-производителей канадской селекции (n = 6), которую использовали для осеменения коров и телок на ферме «Дубровицы» ФГУП «Кленово-Чегодаево». Концентрацию и подвижность сперматозоидов оценивали с помощью компьютерной технологии Computer-assisted semen analysis (CASA). Состояние хроматина в сперматозоидах изучали методом акридин-оранжевого теста с флуоресцентной микроскопией. Морфологию и целостность акросом оценивали с использованием Дифф-Квик окраски. Всего на ферме «Дубровицы» плодотворно были осеменены 161 корова и 29 телок, от них получены 180 телят, из них 97 бычков и 83 телки. Установлено, что одним из биотических факторов, влияющих на репродуктивные показатели, является состояние ДНК в хроматине. От степени фрагментации ДНК в хроматине сперматозоидов зависит их оплодотворяющая способность и другие показатели фертильности быков-производителей. Уровень воспроизведения обусловлен репродуктивным потенциалом как коров, так и быков-производителей. Для комплексной оценки качества спермы быков-производителей необходимо использовать компьютерные технологии с привлечением максимального числа показателей, в том числе наследственного материала, содержащегося в сперматозоидах. Целесообразно при оценке быков-производителей вести учет благополучия отелов, выбытия коров после патологических отелов, абортов, мертворождаемости. Благополучие отелов способствует увеличению сроков последующего производственного использования коров, осемененных спермой быков-производителей, характеризующихся легкими отелами. Для достоверной оценки биологической полноценности спермы необходимо использовать дополнительный показатель, а именно степень фрагментации ДНК в хроматине. У быков-производителей с высокой степенью фрагментации ДНК получены низкие результаты оплодотворяемости.

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Post-Thaw Sperm Quality and Functionality in the Autochthonous Pig Breed Gochu Asturcelta

Animals ◽

10.3390/ani11071885 ◽

2021 ◽

Vol 11 (7) ◽

pp. 1885

Author(s):

José Néstor Caamaño ◽

Carolina Tamargo ◽

Inmaculada Parrilla ◽

Felipe Martínez-Pastor ◽

Lorena Padilla ◽

...

Keyword(s):

Genetic Resource ◽

Sperm Quality ◽

Genetic Material ◽

Mitochondrial Activity ◽

Computer Assisted ◽

Kinematic Parameters ◽

Linear Mixed Effects ◽

Sperm Analysis ◽

Computer Assisted Sperm Analysis ◽

Analysis System

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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