scholarly journals Evaluation of porcine semen quality by portable and desktop CASA systems – Short communication

2020 ◽  
Vol 68 (2) ◽  
pp. 197-199
Author(s):  
Eszter Balogh ◽  
Anna Boglárka Dálnoki ◽  
László Rózsa ◽  
Viktória Johanna Debnár ◽  
Orsolya Varga-Balogh ◽  
...  

AbstractWhen using artificial insemination in porcine reproduction, one of the most important requirements is the suitable quality of semen regarding its total motility (TM) and progressive motility (PM). Computer-assisted sperm analysis (CASA) is an appropriate method to analyse the quality of semen. Recently a portable instrument has been developed to help specialists in their everyday field work. In our study, semen quality was measured simultaneously by the portable device (Ongo) and a laboratory CASA system (Microptic) to compare TM and PM values obtained by these appliances at a concentration of 50 × 106 spermatozoa/mL. Agreement between measurements was evaluated with a Bland-Altman plot. Strong correlation was found between the investigated instruments for all the three parameters, i.e. sperm concentration, TM and PM. However, a few measurements fell outside the defined range of acceptance.

2020 ◽  
Author(s):  
Rafael Suárez Mesa ◽  
Joan Estany ◽  
Iang Schroniltgen Rondón-Barragán

Abstract Background Colombian Creole pigs are a valuable resource for rural livelihood and research. There are three officially recognized Creole breeds in Colombia (Zungo, ZU; Casco de Mula, CM; and San Pedreño, SP). The phenotypic characterization of these breeds is still very limited, including the reproductive performance of their boars, which is a key issue for developing conservation and dissemination strategies. The aim of this research was to assess the semen quality of Colombian Creole breeds as compared to commonly used international breeds. Results Seven boars for each Colombian Creole (ZU, CM, and SP) and international (Duroc, Belgian Landrace, and Pietrain) breeds were used in the experiment. Two doses of semen per boar were assessed in duplicate for sperm kinetics and membrane and acrosome integrity using computer-assisted sperm analysis and flow cytometry, respectively. On average, the Creole pigs, as compared to international breeds, showed lower (P<0.05) volumeof fluid ejaculated(185.5 mL vs 239.9 mL) as well as sperm concentration (340.5 vs to 395.4, in million sperm/mL), motility (90.9% vs 95.3%) and progressive motility (63.1% vs 67.2%). No relevant differences between breeds for sperm velocity traits were observed, but Creole pigs had lower (P<0.05) proportion of morphologic normal sperm (86.1% vs 90.6%) and of sperm with intact mitochondria plasma membrane and acrosome (76.8% vs 87.5%). Mitochondrial membrane potential did not differ between Creole and international breeds. These results mean that Creole breeds had 60.5% less normal and motile sperm per ejaculate than international breeds. Amongst Creole breeds, SP had larger ejaculates and ZU showed greater proportion of normal and motile sperm, but they did not differ for the amount of normal and motile sperm per ejaculate. Conclusion The semen of Colombian Creole pigs is acceptable but less abundant and rich in normal and motile spermatozoa than that collected from commercial breeds. This fact should be considered in developing recommendations for semen processing in Creole pigs. Findings provided here can give new impetus to the conservation and insemination of Creole pigs.


Respuestas ◽  
2013 ◽  
Vol 18 (2) ◽  
pp. 16-27
Author(s):  
Leonardo Hernández-Corredor ◽  
Alexander Nivia-Osuna ◽  
Daniel Hernández-Villamizar ◽  
Jorge Alexander Rubio-Parada ◽  
Armando Quintero-Moreno

 El estudio evaluó la motilidad espermática y su efecto postdescongelación en semen caprino, en dos medios comerciales (Andromed® y TwoStep®) y diferentes protocolos de congelación (medio completo, con adicción del 10% de yema de huevo, semen centrifugado y sobrenadante seminal), se utilizaron machos de la raza alpina de la Universidad Francisco de Paula Santander Ocaña, el semen fue colectado con electroeyaculador, una vez los medios terminados y parte de los contenidos seminales enteros o centrifugados mezclados, se estabilizó por 2 horas, se envasó en pajillas de 0,5 cc y se congela en vapores de nitrógeno por 10 minutos, las pajillas se llevaron al laboratorio de Andrología de la Universidad del Zulia y por medio del sistema C.A.S.A.(Computer Assisted Sperm Análisis) se evaluaron los parámetros de motilidad como velocidad curvilínea (VCL), velocidad rectilínea (VSL), velocidad lineal (VAP), índice de linealidad (LIN), índice de rectitud (STR), índice de oscilación (ALH), Amplitud media del desplazamiento lateral de la cabeza del espermatozoide (BCF), los datos fueron analizados por medio del procedimiento GLM de SAS versión 9.0; los mejores índices de motilidad (VCL, ALH, BCF) fueron expresados enel tratamiento de contenido seminal centrifugado en medio Andromed®. (p≤0,001))La mejor progresividad espermática (VSL,LIN,STR)se presentó el tratamiento de Semen completo de caprino, criopreservado en medio comercial TwoStep®. ABSTRACT  The study evaluated the effect sperm motility and sperm post-thawing in goats, two commercial means (Andromed ® and Two Step ®) and different freezing protocols (complete medium with 10% addition of the egg yolk, semen centrifuged supernatant and seminal ), we used males of the Alpine race of the University Francisco de Paula Santander Ocaña, semen was collected with electroejaculator once finished media and part of the whole and centrifuged seminal contents mixed, stabilized by two hours, packed in 0.5 cc straws and frozen in nitrogen vapor for 10 min, the straws were taken to the laboratory of Andrology at the University of Zulia and through CASA system (Computer Assisted Sperm Analysis) were evaluated motility parameters such as curvilinear velocity (VCL), straight line velocity (VSL), linear velocity (VAP), linearity index (LIN), straightness index (STR) Oscillation Index (ALH ) average amplitude of the lateral displacement of the sperm head (BCF), the data were analyzed by the GLM procedure of SAS version 9.0, the highest rates of motility (VCL, ALH, BCF) were expressed in the treatment of seminal content centrifugation Andromed ® medium. (p ≤ 0.001)) The best progressive sperm (VSL, LIN, STR) will present the full Semen treatment goats, cryopreserved at Two Step ® commercial medium.Keywords: semen, buck, Andromed, Two step.


2020 ◽  
Vol 32 (2) ◽  
pp. 137
Author(s):  
Y. Pirosanto ◽  
M. Valera ◽  
A. Molina ◽  
J. Dorado ◽  
S. Demyda-Peyrás

Inbreeding depression, a genetic condition produced by the mating of close-related individuals, has been associated with a reduction of fertility in several species. However, a loss in sperm quality was also associated with age. In horses, the few existing reports have described a tendency of both parameters to produce a negative effect on sperm quality. However, those reports were performed using a subjective evaluation of sperm motility. In the present study, a total of 692 ejaculates from 86 Pure Spanish stallions (PRE), aged between 3 and 22 years, were evaluated using a computer-assisted methodology to determine the effect of inbreeding in four semen parameters: free-gel volume (V), sperm concentration (C, by haemocytometer), and total (TM) and progressive (PM) sperm motility (by Spermvision sperm class analyser; Minitube). The inbreeding coefficient (F) was estimated using 300 000 PRE pedigree records approximately (minimum pedigree depth, eight equivalent complete generations; range, between 1 and 30.1%). Stallion, age, ejaculate, and season of semen collection were the variables included in the statistical model (general linear model), with ejaculate and season being the variables with a major effect (by variance components analysis). Our results showed that sperm concentration (r=−0.18; P&lt;0.0001) and volume (to a lesser extent) were reduced with advancing age, both showing a major decline after 15 years of age. To the contrary, sperm motility was not affected by age of the stallion. We also found a negative correlation between the inbreeding coefficient and ejaculate volume (r=−0.14; P&lt;0.001), with a marked decrease seen when F was between 7 and 20%. Also, a negative correlation was observed in PM (r=−0.08; P&lt;0.05), although to a lower extent. Conversely, C and TM were not affected by inbreeding depression (P&gt;0.05). In conclusion, our results demonstrated that high levels of inbreeding can compromise severely the sperm quality of the PRE stallion, which, subsequently, may have a negative influence on fertility. Ongoing studies using genomic data will help to detect genetic variants associated with stallion semen quality and how it is influenced by inbreeding in specific genomic regions.


2017 ◽  
Vol 62 (No. 6) ◽  
pp. 227-233 ◽  
Author(s):  
J. Šichtař ◽  
A. Nehasilová ◽  
O. Šimoník ◽  
F. Bubeníčková

The aim of the study was to evaluate the effect of two different extenders on sperm characteristic before equilibration and post-thaw in the endangered Old Kladruber stallions. Also, the response of individual stallions to the extenders used was tested. Semen was collected from six stallions every other day within one week. After centrifugation of the collected sperm-rich fraction, the supernatant was removed and sperm pellets were divided to two aliquots; these were diluted either with Gent (Minitube, Germany) or privately manufactured lactose-EDTA-egg yolk extender (Lact). Three cryopreserved insemination doses (IDs) from each extender (Gent and Lact) were prepared for each stallion from one collection (108 samples from six stallions in total). As a parameter of quality, the motility (computer assisted sperm analysis), viability (fluorescence staining), and morphology (eosin/nigrosine staining) were evaluated after dilution with freezing extenders (fresh) and after thawing (frozen-thawed). The different effects of chosen extenders on the quality of fresh semen were only manifested in higher kinematic parameters of sperm when the Lact extender was used. However, in frozen-thawed samples, the Gent extender yielded significantly better results in all of the evaluated parameters. The representation of sperm subpopulation was significantly influenced by extender in fresh as well as frozen-thawed samples; moreover, we found a significant effect of freezing on the distribution of these subpopulations. The response of individual stallions to chosen extenders was evident in the different quality of fresh as well as frozen-thawed IDs; Gent extender yielded better frozen-thawed IDs. Based on our results, among others describing quality parameters of ejaculate in endangered Old Kladruber stallions, we can recommend using Gent extender for the production of frozen-thawed IDs.  


2013 ◽  
Vol 25 (8) ◽  
pp. 1134 ◽  
Author(s):  
Aimee J. Silla

Anurans exhibit a greater reproductive diversity than any other vertebrate order. However, studies investigating the effects of the external fertilisation environment on fertilisation success are limited to aquatic-breeding species. This study investigated the effects of fertilisation medium osmolality, sperm concentration and short-term oocyte storage on fertilisation success in a terrestrial-breeding anuran, Pseudophryne guentheri. Split-clutch experimental designs were used to determine optimal fertilisation conditions. To determine the effect of short-term sperm storage, sperm viability was assessed using fluorescence microscopy and percentage sperm motility and velocity quantified with a computer-assisted sperm analysis system. Fertilisation success was highest in media ranging in osmolality from 25 mOsm kg–1 to 100 mOsm kg–1, representing a broader range and higher optimal osmolality than previously reported for aquatic breeders. High rates of fertilisation (>75%) were achieved in relatively low sperm concentrations (2.5 × 104 mL–1). Oocytes stored in isotonic solutions (200 mOsm kg–1) retained fertilisation capacity (32%) after 8 h of storage, while sperm suspensions maintained motility (≥26%) for 13 days. Additional studies on terrestrial-breeding anurans will be required to ascertain whether the optimal fertilisation conditions reported reflect adaptations to achieve fertilisation in a terrestrial environment.


Author(s):  
Gemma Gaitskell-Phillips ◽  
Francisco E Martín-Cano ◽  
José M Ortiz-Rodríguez ◽  
Antonio Silva-Rodríguez ◽  
Maria C Gil ◽  
...  

Abstract The identification of stallions and or ejaculates that will provide commercially acceptable quality post-thaw before cryopreservation is of great interest, avoiding wasting time and resources freezing ejaculates that will not achieve sufficient quality to be marketed. Our hypothesis was that after bioinformatic analysis, the study of the stallion sperm proteome can provide discriminant variables able to predict the post-thaw quality of the ejaculate. At least three ejaculates from 10 different stallions were frozen following a split sample design. Half of the ejaculate was analyzed as a fresh aliquot and the other half was frozen and then analyzed as a frozen-thawed aliquot. Computer-assisted sperm analysis and flow cytometry were used to analyze sperm quality. Detailed proteomic analysis was performed on fresh and frozen and thawed aliquots, and bioinformatic analysis was used to identify discriminant variables in fresh samples able to predict the outcome of cryopreservation. Those with a fold change &gt; 3, a P = 8.2e-04, and a q = 0.074 (equivalent to False discovery rate (FDR)) were selected, and the following proteins were identified in fresh samples as discriminant variables of good motility post-thaw: F6YTG8, K9K273, A0A3Q2I7V9, F7CE45, F6YU15, and F6SKR3. Other discriminant variables were also identified as predictors of good mitochondrial membrane potential and viability post-thaw. We concluded that proteomic approaches are a powerful tool to improve current sperm biotechnologies.


2008 ◽  
Vol 20 (1) ◽  
pp. 87
Author(s):  
C. O. Hidalgo ◽  
A. Rodríguez ◽  
C. Díez ◽  
D. Martín ◽  
M. Carbajo ◽  
...  

The Bermeya goats are an endangered autochthonous breed distributed in the north of Spain. To ensure their genetic diversity and long-term survival, morphological and functional parameters of the semen must be known in order to preserve the current genetic stock in a germplasm bank. The aim of this work was to establish basic characteristics and post-thaw survival of Bermeya goat's semen obtained by electro-ejaculation, that is not well described in the literature. The semen was collected by electro-ejaculation from 7 bucks, 1 to 7 years old, twice per week, for 9 weeks (n = 83). Fresh semen was evaluated for volume (V), concentration (C), motility, morphology, functional integrity of the sperm (spz) membranes (hypoosmotic swelling test; HOST), and acrosome integrity rate (NAR). Individual and progressive sperm motility were analyzed by means of a computer-assisted sperm analysis system (CASA: SCA 2002�, Microptic, Barcelona, Spain) immediately after dilution with the extender at 37�C, and after cooling to 4�C; five fields per sample (diluted to 204 � 106 spz mL–1) were evaluated under a phase contrast microscope (100�). The NAR and morphological abnormalities of sperm head, midpiece, tail, and cytoplasmic droplets were determined by counting 100 spz under 1000�. For freezing, ejaculates with at least 80% motile spz were diluted at 32�C with Krebs-Ringer solution containing 20% egg yolk and 14% glycerol to a final concentration of 400 � 106 spz mL–1, cooled to 4�C for 90 min, aspirated into 0.25-mL plastic straws (IMV�, L'Aigle, France), frozen at 7 cm above liquid nitrogen (LN2) phase for 10 min, and then plunged into the LN2. Straws were thawed in a water bath at 39�C for 30 s for post-thaw survival analysis. Data were analyzed by the GLM and FREQ procedures (SAS; SAS Institute, Inc., Cary, NC, USA) and expressed as means � standard error. Fresh semen characteristics were: V = 1.7 � 0.1 mL; C = 2619 � 106 � 153 spz mL–1; total and progressive motility were 89.0 � 2.1% and 66.9 � 2.1%, respectively. Percentages of head abnormalities were 4.8 � 0.5; midpiece: 3.8 � 0.7; tail: 4.7 � 1.0; cytoplasmic droplets: 8.3 � 0.7; intact acrosome: 91.8 � 0.6; and membrane integrity: 49.2 � 2.1. At 4�C, the % of total motile spz was 62.6 � 1.6, and the post-thaw survival rate was 46.3 � 1.5. There were only individual differences (P < 0.001) between bucks on sperm concentration, head abnormalities, and cytoplasmic droplets. In conclusion, our results indicate that semen quality is related to each individual animal and that electro-ejaculation allows collection of semen of satisfactory quality to use as fresh and for cryopreservation. However, the validity of our results for possible future sperm banking of endangered Bermeya goats semen must be confirmed by field trials.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 48-49
Author(s):  
Elena Nikitkina ◽  
Artem Musidray ◽  
Anna Krutikova ◽  
Svetlana Timofeeva ◽  
Kirill Plemyashov ◽  
...  

Abstract Reindeer herding is a traditional activity of the indigenous peoples of the Arctic. Assisted reproductive technology in reindeer herding are under development. This is due to the difficulty of collecting reindeer sperm (Rangifer tarandus) in the Arctic. The aim of the work was to study the influence of age and rut period on the quality of reindeer (Rangifer tarandus) sperm. Semen was collected in Taimyr in autumn 2017, 2018. Ejaculate volume, total (TM) and progressive motility (PM) and sperm concentration were evaluated. TM, PM and sperm concentration were measured by CASA (computer-assisted semen analysis). Sperm was not found in caudae epididymidae in males at the age of six months. The sperm was collected only from caudae epididymidae from males aged 1.5 and 2.5 years. Collecting of ejaculated semen is possible in males from 3.5 years of age. The volume of ejaculate increased from 0.3±0.09 ml (mean±SD) at the age of 3.5 years to 0.7±0.09 ml at the age of 5.5 years and older (P &lt; 0.05). Concentration of cells increased by 2 times in older males. It is known that reindeer sperm can be collected only during the rut. At the beginning and end of the rut, the collection of sperm by electroejaculation was not always successful, sperm was collected in 3 of 10 cases, while in the middle of the rut in 9 of 10 cases. There was no significant effect of the rut period on the ejaculate volume and sperm concentration. The lowest TM (51.1±2.16%) and PM (33.5±7.90%) were at the beginning of the rut, the highest at the end (73.0±5.02% and 59.0 ± 6,30%) (P &lt; 0.01). Thus, the effect of age and rutting period on the reindeer sperm was observed. Authors acknowledge financial support from Russian Science Foundation, Grant No:17-16-01023.


Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1986
Author(s):  
Sara El Kadili ◽  
Nathalie Kirschvink ◽  
Marianne Raes ◽  
Jean Loup Bister ◽  
Bouchaib Archa ◽  
...  

The study aimed at determining the effect of storage and season on fresh semen of Beni Arouss goats. Ejaculates were collected at monthly intervals from seven mature bucks and were extended at a final concentration of 800 × 106 spermatozoa. ml-1 and stored at 16 °C for 24 h. Semen motility, viability and normal morphology were assessed at 0, 4, 8 and 24 h after collection. Motility and normal morphology parameters were recorded using computer-assisted sperm analysis (CASA) and viability was analyzed using eosin–nigrosin staining. As expected, motility, viability and normal morphology parameters showed a significant reduction within 24 h of storage and during all seasons (p < 0.05). However, semen collected in summer maintained a better quality after 24 h of storage at 16 °C than semen collected during the other periods (p < 0.05). In conclusion, the storage ability of Beni Arouss bucks’ semen stored at 16 °C was significantly higher during the summer.


2013 ◽  
Vol 57 (3) ◽  
pp. 429-432 ◽  
Author(s):  
Anna Domosławska ◽  
Sławomir Zduńczyk ◽  
Wojciech Niżański ◽  
Tomasz Janowski

Abstract Semen quality parameters of infertile and fertile dogs were compared. Sperm concentration and semen motility parameters were measured by the Hamilton-Thorne Semen Analyser IVOS 12.3. The spermatozoal morphology and the percentage of live spermatozoa were examined microscopically. Forty-six dogs of various breeds were examined. Twenty dogs had a conception failure within last year. These dogs had a history of prior normal fertility. Twenty six fertile dogs served as control. All animals underwent a clinical examination as well as ultrasonography. Sperm concentration was significantly lower in infertile dogs than in fertile dogs. For most determined motility parameters (MOT, PMOT, VAP, VSL, VCL, BCF, RAPID, STATIC) significant differences between infertile and fertile dogs were found. The percentage of spermatozoa with normal morphology also significantly differed between both groups. Ultrasonography of the prostate gland and testes revealed no pathological conditions. The testicular degeneration was assumed to be a possible cause of infertility in these dogs. The present study showed that the most sperm CASA motility parameters were significantly lower in infertile dogs in comparison to the fertile ones, and confirmed the usefulness of the Hamilton-Thorne Semen Analyser for a quick and objective analysis of sperm concentration and motility in dogs.


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