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

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.

scholarly journals 2016 Laboratory guidelines for postvasectomy semen analysis: Association of Biomedical Andrologists, the British Andrology Society and the British Association of Urological Surgeons

2016 ◽  
Vol 69 (7) ◽  
pp. 655-660 ◽  
Author(s):  
P Hancock ◽  
B J Woodward ◽  
A Muneer ◽  
J C Kirkman-Brown
Keyword(s):  
United Kingdom ◽  
General Practitioner ◽  
Semen Analysis ◽  
Single Sample ◽  
British Association ◽  
Motile Sperm ◽  
Iso 15189 ◽  
The United Kingdom ◽  
Two Samples ◽  
The Uk

Post-vasectomy semen analysis (PVSA) is the procedure used to establish whether sperm are present in the semen following a vasectomy. PVSA is presently carried out by a wide variety of individuals, ranging from doctors and nurses in general practitioner (GP) surgeries to specialist scientists in andrology laboratories, with highly variable results.Key recommendations are that: (1) PVSA should take place a minimum of 12 weeks after surgery and after a minimum of 20 ejaculations. (2) Laboratories should routinely examine samples within 4 h of production if assessing for the presence of sperm. If non-motile sperm are observed, further samples must be examined within 1 h of production. (3) Assessment of a single sample is acceptable to confirm vasectomy success if all recommendations and laboratory methodology are met and no sperm are observed. Clearance can then be given. (4) The level for special clearance should be <100 000/mL non-motile sperm. Special clearance cannot be provided if any motile sperm are observed and should only be given after assessment of two samples in full accordance with the methods contained within these guidelines. Surgeons are responsible both preoperatively and postoperatively for the counselling of patients and their partners regarding complications and the possibility of late recanalisation after clearance. These 2016 guidelines replace the 2002 British Andrology Society (BAS) laboratory guidelines and should be regarded as definitive for the UK in the provision of a quality PVSA service, accredited to ISO 15189:2012, as overseen by the United Kingdom Accreditation Service (UKAS).

scholarly journals Effects of Apigenin and Astragalus Polysaccharide on the Cryopreservation of Bull Semen

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1506
Author(s):  
Hongtao Wang ◽  
Ping Lu ◽  
Chongshan Yuan ◽  
Jing Zhao ◽  
Hongyu Liu ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Computer Assisted ◽  
Astragalus Polysaccharides ◽  
Bovine Semen ◽  
Bull Semen ◽  
Straight Line ◽  
Frozen Semen ◽  
Path Distance ◽  
Straight Line Distance

The purpose of this study was to determine the effects of apigenin and astragalus polysaccharides on the cryopreservation of bovine semen. Apigenin, astragalus polysaccharides, or their combination were added to a frozen diluent of bovine semen. Afterwards, Computer Assisted Semen Analysis (CASA), membrane functionality, acrosome integrity, mitochondrial integrity, CAT, SOD, GSH-Px, MDA, and ROS detection were conducted. The results showed that adding 0.2 mmol/L AP or 0.5 mg/mL APS could improve the quality of frozen sperm. Compared to 0.2 mmol/L AP alone, the combination of 0.2 mmol/L AP and 0.3 mg/mL APS significantly increased the total motility (TM), average path distance (DAP), straight line distance (DSL), average path velocity (VAP), curvilinear velocity (VCL), wobble (WOB), and sperm CAT and SOD levels (p < 0.05), while reducing the ROS and MDA levels (p < 0.05). These results indicated that the addition of 0.2 mmol/L AP or 0.5 mg/mL APS alone has a protective effect on the freezing of bovine semen. Compared to the addition of 0.2 mmol/L AP, a combination of 0.2 mmol/L AP and 0.3 mg/mL APS could further improve the quality of frozen semen.

scholarly journals The potential of sperm bovine protamine as a protein marker of semen production and quality at the National Artificial Insemination Center of Indonesia

2021 ◽  
pp. 2473-2481
Author(s):  
Berlin Pandapotan Pardede ◽  
Tulus Maulana ◽  
Ekayanti Mulyawati Kaiin ◽  
Muhammad Agil ◽  
Ni Wayan Kurniani Karja ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Artificial Insemination ◽  
Semen Quality ◽  
Semen Analysis ◽  
Normal Function ◽  
Computer Assisted ◽  
Major Protein ◽  
Protein Marker ◽  
Acridine Orange Staining ◽  
Frozen Semen

Background and Aim: Protamine (PRM) is the major protein in the sperm nucleus and plays an essential role in its normal function. Moreover, PRM has great potential as a protein marker of semen production and quality. This study aimed to assess the potential of sperm bovine PRM as a protein marker of semen production and quality in bulls at the National Artificial Insemination (AI) Center of Indonesia. Materials and Methods: The semen production capacity of each bull was collected from frozen semen production data at the Singosari AI Center for 6 months, and was then divided into two groups (high and low). A total of 440 frozen semen straws from six Limousin (LIM), six Friesian Holstein (FH), six Peranakan Ongole (PO), and four Aceh bulls aged 4-5 years were used in the study. The frozen semen was used to measure the concentration of PRM1, PRM2, and PRM3 using the enzyme immunoassay method. The frozen semen was also used to assess the quality of the semen, including progressive motility (PM) through computer-assisted semen analysis, sperm viability through eosin–nigrosin analysis, and the DNA fragmentation index through Acridine Orange staining. Results: PRM1 was significantly higher in all bull breeds included in the study (p<0.00), followed by PRM2 (p<0.00) and PRM3 (p<0.00). PRM1 significantly affected semen production in LIM, FH, PO, and Aceh bulls (p<0.05). Moreover, PRM2 significantly affected semen production only in FH and Aceh bulls (p<0.05), whereas PRM3 affected this parameter in PO and Aceh bulls exclusively (p<0.05). Consistently and significantly, PRM1 was positively correlated with the PM and viability of sperm and negatively associated with its DNA fragmentation in LIM, FH, PO, and Aceh bulls (p<0.05; p<0.01). The correlation analysis between PRM2 and PRM3 and semen quality parameters varied across all bull breeds; some were positively and negatively correlated (p<0.05; p<0.01), and some were not correlated at all. Conclusion: PRM1 has excellent potential as a protein marker of semen production and quality in bulls at the National AI Center of Indonesia.

120 Use of fixable dyes to analyze equine sperm membrane integrity and acrosome reaction after A23187 treatment

2020 ◽  
Vol 32 (2) ◽  
pp. 187
Author(s):  
I. Ortiz ◽  
M. Felix ◽  
H. Resende ◽  
C. Love ◽  
K. Hinrichs
Keyword(s):  
Flow Cytometry ◽  
Repeated Measures ◽  
Semen Analysis ◽  
Flow Cytometric Analysis ◽  
Membrane Integrity ◽  
Fluorescein Isothiocyanate ◽  
Computer Assisted ◽  
Cell Agglutination ◽  
Staining Protocol ◽  
Viable Sperm

Conventional IVF is not successful in the horse, and current work is focused on factors affecting sperm capacitation in this species. Challenges arise in assessing equine sperm incubated in media containing capacitation promotors, as some of these factors cause sperm head-to-head binding (aggregation). Our preliminary microscopic findings showed that sperm aggregates are largely of viable sperm, whereas nonviable sperm individualize. Thus, data obtained using technologies that analyse only individual cells and gate out aggregates, such as flow cytometry, may not accurately represent the study population. We developed a fixable live/dead/acrosome staining protocol (LD-PSA) that minimizes sperm aggregation. The aim of this study was to evaluate the percentage of viable and acrosome-reacted equine sperm after A23187 treatment, using either LD-PSA or a standard staining protocol (PI-PSA). Sperm from 9 ejaculates were suspended in Hanks’ balanced salt solution (HBSS) and exposed for 10min at 37°C to vehicle (V) or to 10 µM A23187 (C10). The sperm were washed, resuspended in HBSS medium with added lactate and pyruvate and containing 7mgmL−1 of bovine serum albumin (BSA), and assessed immediately (0h) or incubated at 37°C for 2h (the period needed for equine sperm to respond to A23187). Motility was analysed using computer-assisted semen analysis. Each treatment was stained by PI-PSA: propidium iodide and fluorescein isothiocyanate-Pisum sativum agglutinin (PSA) in DPBS; and by LD-PSA: Live/Dead Fixable Red, paraformaldehyde 2%, Triton×1%, and FITC-PSA in Dulbecco's phosphate-buffered saline with Accumax (Stem Cell Technologies), a commercial proprietary cell agglutination inhibitor, before flow cytometric analysis. Differences were analysed using repeated-measures two-way ANOVA. The% total motile sperm (TMOT) for V and C10 treatments were 76.3±3.0 and 71.2±4.7 at 0h (P&gt;0.05), and 70.5±14.8 and 2.4±0.8 at 2h (P&lt;0.05). On flow cytometry, the percentage of events outside the gate for V sperm (0h and 2h combined) was 31.9% in PI-PSA and 21.9% in LD-PSA samples (P&lt;0.01). Measured viability in V samples was significantly lower when stained with PI-PSA than with LD-PSA at 0h (49.2±4.6 vs. 67.1±4.9) and tended to be lower (P=0.07) at 2h (44.0±4.9 vs. 55.1±2.8). Notably, the viability recorded in PI-PSA was 26 percentage points lower than was the TMOT at both 0h and 2h, indicating nonrepresentative results, as nonviable sperm should not be motile. By LD-PSA, this difference was 9 points at 0h and 15 points at 2h. Vehicle sperm showed significantly higher AR values in PI-PSA than in LD-PSA at 0h (30.4±4.0 vs. 17.7±2.4) and 2h (41.9±4.5 vs. 24.0±1.8), as did C10 sperm at 0h (28.9±2.7 vs. 18.0±2.5). The lower values for viability than total motility likely reflect agglutination of viable sperm and thus their exclusion from analysis on flow cytometry. The anti-clumping measures employed in the LD-PSA protocol were associated with increased correspondence of measured viability with TMOT. Thus, LD-PSA may offer a more accurate technique to assess viability and acrosome status of equine sperm incubated in capacitating conditions.

scholarly journals Communicating the “Race” for the COVID-19 Vaccine: An Exploratory Study in Newspapers in the United States, the United Kingdom, and Brazil

2021 ◽  
Vol 6 ◽  
Author(s):  
Luisa Massarani ◽  
Luiz Felipe Fernandes Neves
Keyword(s):  
United States ◽  
New York ◽  
United Kingdom ◽  
Media Coverage ◽  
Exploratory Study ◽  
New York Times ◽  
The United States ◽  
Computer Assisted ◽  
The United Kingdom ◽  
Understanding Of Science

The search for an effective solution to control the COVID-19 pandemic has mobilized an unprecedented effort by science to develop a vaccine against the disease, in which pharmaceutical companies and scientific institutions from several countries participate. The world closely monitors research in this area, especially through media coverage, which plays a key role in the dissemination of trustful information and in the public’s understanding of science and health. On the other hand, anti-vaccine movements dispute space in this communication environment, which raises concerns of the authorities regarding the willingness of the population to get vaccinated. In this exploratory study, we used computer-assisted content analysis techniques, with WordStat software, to identify the most addressed terms, semantic clusters, actors, institutions, and countries in the texts and titles of 716 articles on the COVID-19 vaccine, published by The New York Times (US), The Guardian (United Kingdom), and Folha de São Paulo (Brazil), from January to October 2020. We sought to analyze similarities and differences of countries that stood out by the science denialism stance of their government leaders, reflecting on the severity of the pandemic in these places. Our results indicate that each newspaper emphasized the potential vaccines developed by laboratories in their countries or that have established partnerships with national institutions, but with a more politicized approach in Brazil and a little more technical-scientific approach in the United States and the United Kingdom. In external issues, the newspapers characterized the search for the discovery of a vaccine as a race in which nations and blocs historically marked by economic, political, and ideological disputes are competing, such as the United States, Europe, China, and Russia. The results lead us to reflect on the responsibility of the media to not only inform correctly but also not to create stigmas related to the origin of the vaccine and combat misinformation.

105 Increasing fertility of interspecific hybrid males using biotechnological approaches

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.

scholarly journals 12PREDICTION OF BULL FERTILITY BY COMPUTER ASSISTED SEMEN ANALYSIS

2004 ◽  
Vol 16 (2) ◽  
pp. 128 ◽  
Author(s):  
S. Cseh ◽  
T. Polichronopoulos ◽  
L. Solti
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Reproductive Process ◽  
Return Rates ◽  
Frozen Semen ◽  
Chi Square Analysis ◽  
Fertilizing Capability ◽  
Weighted Values

Sperm motility is clearly essential for fertilization both in vivo and in vitro. Motility is necessary for successful sperm transport, a step that is bypassed with in vitro fertilization. Recently, increasing attention has been paid to the objective evaluation and characterization of sperm motility more than simply determining the total proportion of motile spermatozoa. The purpose of computerassisted semen analysis (CASA) is to provide values for sperm concentration and sperm motility more rapidly and accurately than those obtained with traditional semen analyses methods. The objective of our experiment was to investigate the effect of specific aspects of sperm movement, such as the velocity of progression and the actual pattern of movement, to the fertilizing capability of sperm. Frozen semen samples of 10 HF breeding bulls were used in the study. For the motility analyses, Medealab CASA system (Medealab, Germany, Ver. 4.1) was used, and the velocity parameter of VCL (curvalinear velocity, μms−1), VSL (straight line velocity, μms−1), and VAP (average path velocity, μms−1) were evaluated and compared with the Day 30 and 75 non−return rates (NR30 and NR75). For every sample, a total of 10 fields were examined for 8s using a disposable 20 micron capillary chamber (CellVision, USA) giving a total of 1165 to 2831 cells evaluated. Chi square analysis, analyses of variance and linear correlation coefficient was applied to the statistical evaluation and comparison of the results. Data are based on weighted values. From the same batch of the analyzed frozen semen, a total of 8099 females were inseminated in more than 100 farms with a total of 6590 animals being positive for pregnancy at Day 30 and 4525 animals at Day 75. Within the bulls, differences were found in the values of NR30 and NR75 (P&lt;0.05). Our data indicate very strong differences between the males’ NR30 and NR75 values (NR30: 65.6%±13.04 to 79.6%±11.17; P&lt;0.001 and NR75: 37.8%±10.38 to 58.3%±15.53; P&lt;0.001) reflecting the individual differences in the fertilizing capability of the males. All velocity parameters show very high correlation with strong significance both non−return rates but the best values belong to VAP (NR30 and NR75; P&lt;0.02). Our data indicate that the bulls with lower VCL (25.51±33.04 to 79.54±58.03), VSL (11.35±19.45 to 36.36±35.71), and VAP (12.67±19.06 to 41.75±34.45) values showed lower fertilization rates both at NR30 and NR75. Computer and video technologies have advanced rapidly in recent years; thus the capability and accuracy of the latest versions of CASA systems are considerably better and they give more information about the different motion characteristics of spermatozoa. Because of the vital role of sperm motility in the reproductive process, such systems will enable us to move into a new era of diagnostic andrology and predict the fertilizing capability of semen. Supported by NKFP-Grants 4/040/2001 and 4/031/2001.

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.

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