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Keywords = canine spermatozoa

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14 pages, 268 KiB  
Article
Stability over Time of the Sperm Motility Biomarker proAKAP4 in Repeated Dog Ejaculates
by Giulia Siena, Alain Fontbonne, Barbara Contiero, Cindy Maenhoudt, Guillaume Robiteau, Sarah Slimani, Nicolas Sergeant, Laurent Tiret and Chiara Milani
Animals 2025, 15(8), 1160; https://doi.org/10.3390/ani15081160 - 17 Apr 2025
Viewed by 586
Abstract
ProAKAP4 is a sperm structural protein involved in motility, capacitation, and fertility. Previous studies suggested it as a suitable marker for canine sperm evaluation. Our study aimed to assess proAKAP4 concentration variations among different ejaculates collected from the same dogs. Fourteen male dogs [...] Read more.
ProAKAP4 is a sperm structural protein involved in motility, capacitation, and fertility. Previous studies suggested it as a suitable marker for canine sperm evaluation. Our study aimed to assess proAKAP4 concentration variations among different ejaculates collected from the same dogs. Fourteen male dogs from 12 different breeds, 1–14 years old and 6.9–95 kg bodyweight, underwent semen collection at least twice (1–4 times) during the same or different days. Sperm concentration, morphology, proAKAP4 concentration, total (TM%), and progressive motility (PM%) were considered. ProAKAP4 values were divided into four classes: ≤15, 15–40, 40–60, and >60 ng/10 M spermatozoa. The time interval between two semen collections was classified as: first collection (T0), semen collection performed 2–3 h after the first one (T0.5) and after ≥1 day (1–36 days, T1). Thirty-three ejaculates were collected. A correlation was found between proAKAP4 classes and TM% (r = 0.40, p = 0.049), whereas no correlation was found neither between proAKAP4 and sperm morphology nor for proAKAP4 classes between collection times (T0 vs. T0.5, p = 0.655; T0 vs. T1, p = 0.564). Our results confirm the correlation between proAKAP4 and TM% in dog semen. ProAKAP4 concentration appears stable in different ejaculates collected from the same dogs. Further studies are needed on the relationship between proAKAP4 and other canine semen parameters and fertility outcomes. Full article
10 pages, 798 KiB  
Article
The Interaction between Canine Semen Bacteria and Semen Quality Parameters
by Šarūnė Sorkytė, Rita Šiugždinienė, Marius Virgailis, Gintarė Vaičiulienė, Anna Wysokińska, Ewa Wójcik, Paulius Matusevičius, Audronė Rekešiūtė and Neringa Sutkevičienė
Animals 2024, 14(15), 2151; https://doi.org/10.3390/ani14152151 - 24 Jul 2024
Viewed by 1720
Abstract
Assessing canine semen quality helps to detect infertility in males, but identifying factors that influence canine semen quality is a complicated task. The objective of this study was the assessment of the potential influence of bacteria found in canine semen samples on the [...] Read more.
Assessing canine semen quality helps to detect infertility in males, but identifying factors that influence canine semen quality is a complicated task. The objective of this study was the assessment of the potential influence of bacteria found in canine semen samples on the characteristics of dogs’ semen. In this study, semen samples were collected manually from 30 dogs and subjected to a comprehensive examination. The results of sperm motility, concentration, viability, and morphology were statistically analysed in relation to the number of bacteria in the semen (CFUs/mL) and the seminal microbiota. Samples with an increased bacterial count per millilitre were associated with lower-quality sperm motility (p < 0.05). The most frequently isolated bacterial genera from the analysed semen samples were Staphylococcus spp. (26.0%), Corynebacterium spp. (17.8%), and Streptococcus spp. (16.4%). The presence of β-haemolytic Escherichia coli bacteria was linked to suboptimal semen samples, characterised by significantly reduced semen viability and a lower proportion of morphologically normal spermatozoa (p < 0.05). Corynebacterium spp. was associated with reduced bacterial load and superior semen quality (p < 0.01). These findings highlight the importance of bacterial cell counts and microbiota diversity in relation to various factors influencing canine semen quality, providing a more comprehensive understanding of canine reproductive well-being. Full article
(This article belongs to the Special Issue Animal Reproduction: Semen Quality Assessment, Volume II)
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24 pages, 9566 KiB  
Article
Ultrasonography of Testicular Maturation and Correlation with Body Growth and Semen Evaluation in Beagle Dog Model
by Athina P. Venianaki, Mariana S. Barbagianni, George C. Fthenakis, Apostolos D. Galatos and Pagona G. Gouletsou
Vet. Sci. 2024, 11(6), 270; https://doi.org/10.3390/vetsci11060270 - 14 Jun 2024
Viewed by 2652
Abstract
This prospective study investigated the ultrasonographic appearance of the canine testis from birth to adulthood. Eight purpose-bred laboratory Beagle-breed dogs were monitored from 4 to 40 weeks of life. The following parameters were evaluated every two weeks: bodyweight and height, scrotal and testicular [...] Read more.
This prospective study investigated the ultrasonographic appearance of the canine testis from birth to adulthood. Eight purpose-bred laboratory Beagle-breed dogs were monitored from 4 to 40 weeks of life. The following parameters were evaluated every two weeks: bodyweight and height, scrotal and testicular volume, ultrasonographically measured testicular volume, echogenicity, heterogeneity, blood-flow score, ratio of the grayscale intensity value of the testis to the capsule, ejaculate volume, motility, viability, and number of spermatozoa. A correlation analysis was carried out between the various measurements obtained. Fertility was achieved in the 36th week of life. The echogenicity of the testicular parenchyma increased with age, and subsequently to the 30th week of life remained constant. The heterogeneity of the testicular parenchyma, as was evaluated by the standard deviation of the values of grayscale intensity of the parenchyma, also increased with age and was >19 at the onset of fertility. The ratio of grayscale intensity of testicular parenchyma had values < 200 at maturity. A colour Doppler evaluation first detected blood flow in the testis in the 22nd week. After the 32nd week, distinct signals were visible. In the 36th week, >80% of the testes imaged had visible vessels. A significant correlation was found between all the evaluation methods. The findings of the study may help clinicians detect the onset of fertility in dogs, especially when semen evaluation is not feasible; however, their applicability in all breeds or individuals might possibly vary due to genetic, physiological, and developmental differences. In summary, the study ultrasonographically explores the testicular maturity in dogs, with the aim to improve clinical assessments and health management in these animals. Full article
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13 pages, 6032 KiB  
Article
Extrusion of Neutrophil Extracellular Traps (NETs) Negatively Impacts Canine Sperm Functions: Implications in Reproductive Failure
by Marion León, Claudia Moya, Rodrigo Rivera-Concha, Felipe Pezo, Pamela Uribe, Mabel Schulz, Raúl Sánchez, Anja Taubert, Carlos Hermosilla and Fabiola Zambrano
Int. J. Mol. Sci. 2024, 25(11), 6216; https://doi.org/10.3390/ijms25116216 - 5 Jun 2024
Cited by 3 | Viewed by 1529
Abstract
Reproductive failure in dogs is often due to unknown causes, and correct diagnosis and treatment are not always achieved. This condition is associated with various congenital and acquired etiologies that develop inflammatory processes, causing an increase in the number of leukocytes within the [...] Read more.
Reproductive failure in dogs is often due to unknown causes, and correct diagnosis and treatment are not always achieved. This condition is associated with various congenital and acquired etiologies that develop inflammatory processes, causing an increase in the number of leukocytes within the female reproductive tract (FRT). An encounter between polymorphonuclear neutrophils (PMNs) and infectious agents or inflammation in the FRT could trigger neutrophil extracellular traps (NETs), which are associated with significantly decreased motility and damage to sperm functional parameters in other species, including humans. This study describes the interaction between canine PMNs and spermatozoa and characterizes the release of NETs, in addition to evaluating the consequences of these structures on canine sperm function. To identify and visualize NETs, May–Grünwald Giemsa staining and immunofluorescence for neutrophil elastase (NE) were performed on canine semen samples and sperm/PMN co-cultures. Sperm viability was assessed using SYBR/PI and acrosome integrity was assessed using PNA-FITC/PI by flow cytometry. The results demonstrate NETs release in native semen samples and PMN/sperm co-cultures. In addition, NETs negatively affect canine sperm function parameters. This is the first report on the ability of NETs to efficiently entrap canine spermatozoa, and to provide additional data on the adverse effects of NETs on male gametes. Therefore, NETs formation should be considered in future studies of canine reproductive failure, as these extracellular fibers and NET-derived pro-inflammatory capacities will impede proper oocyte fertilization and embryo implantation. These data will serve as a basis to explain certain reproductive failures of dogs and provide new information about triggers and molecules involved in adverse effects of NETosis for domestic pet animals. Full article
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19 pages, 2954 KiB  
Article
Proteome Profiling of Canine Epididymal Fluid: In Search of Protein Markers of Epididymal Sperm Motility
by Aleksandra W. Cichowska, Jerzy Wisniewski, Mariusz A. Bromke, Beata Olejnik and Marzena Mogielnicka-Brzozowska
Int. J. Mol. Sci. 2023, 24(19), 14790; https://doi.org/10.3390/ijms241914790 - 30 Sep 2023
Cited by 3 | Viewed by 2125
Abstract
Sperm maturation in the epididymis is based on interactions with proteins from epididymal fluid (EF). The aim of the study was to profile canine EF proteome and investigate correlations between EF protein content and epididymal spermatozoa (ES) motion parameters. Twenty-three male dogs were [...] Read more.
Sperm maturation in the epididymis is based on interactions with proteins from epididymal fluid (EF). The aim of the study was to profile canine EF proteome and investigate correlations between EF protein content and epididymal spermatozoa (ES) motion parameters. Twenty-three male dogs were divided into two groups: good sperm motility (GSM) and poor sperm motility (PSM). The total motility and progressive motility differed significantly (p = 0.031; p < 0.001, respectively) between the GSM group and the PSM group. The semen samples were centrifuged to separate the EF apart from the ES. The canine EF proteins were analyzed using nano-liquid chromatography, which was coupled with quadrupole time-of-flight mass spectrometry (NanoUPLC-Q-TOF/MS) and bioinformatic tools for the first time. A total of 915 proteins were identified (GSM—506; PSM—409, respectively). UniProt identification resulted in six unique proteins (UPs) in the GSM group of dogs and four UPs in the PSM group. A semi-quantitative analysis showed a higher abundance (p < 0.05) of four differentially expressed proteins in the GSM group (ALB, CRISP2, LCNL1, PTGDS). Motility-dependent variations were detected in the EF proteome and were related to important metabolic pathways, which might suggest that several proteins could be potential ES motility biomarkers. Full article
(This article belongs to the Special Issue Novel Insights into the Biology of Spermatozoa 2.0)
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14 pages, 2255 KiB  
Article
Does Better Post-Thaw Motility of Dog Sperm Frozen with CLC Mean Better Zona Pellucida Binding Ability?
by Zuzanna Ligocka, Agnieszka Partyka, Sabine Schäfer-Somi, Anna Mucha and Wojciech Niżański
Animals 2023, 13(10), 1580; https://doi.org/10.3390/ani13101580 - 9 May 2023
Cited by 1 | Viewed by 2514
Abstract
Even though the search for methods improving cryopreservation of canine spermatozoa led to an improvement of post-thaw quality, fertilizing results after insemination with frozen–thawed semen are still not satisfying. In this study, we focused on modification of spermatozoa membrane fluidity and investigated whether [...] Read more.
Even though the search for methods improving cryopreservation of canine spermatozoa led to an improvement of post-thaw quality, fertilizing results after insemination with frozen–thawed semen are still not satisfying. In this study, we focused on modification of spermatozoa membrane fluidity and investigated whether kinematic parameters as assessed by computer-assisted semen analyzer (CASA) can be improved. The primary aim of our study was to investigate whether the use of cholesterol-loaded cyclodextrins (CLC; 0.5 mg, 1 mg, 2 mg) and 2-Hydroxypropyl-ß-cyclodextrin (HBCD; 1 mg) positively influence capacitation status as examined by tyrosinphosphorylation, cholesterol efflux and zona binding assay (ZBA) of spermatozoa. The use of 0.5 mg of CLC increased the percentage of motile, progressive and rapid spermatozoa compared to the control. Addition of HBCD decreased motility and progressive motility of spermatozoa and the population with rapid movement in comparison to the control. The percentage of live spermatozoa without efflux of cholesterol compared to the control was increased when extender with 0.5 mg of CLC was used. There was no change in capacitation status. The zona binding ability of spermatozoa was significantly lower in the group with 0.5 mg of CLC than in the control. In conclusion, these results suggest that improvement of kinematic parameters does not necessarily coincide with better zona pellucida binding ability of spermatozoa. Full article
(This article belongs to the Section Companion Animals)
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15 pages, 2306 KiB  
Article
Defining an Optimal Range of Centrifugation Parameters for Canine Semen Processing
by Nicole Sugai, Stephen Werre, Julie Cecere and Orsolya Balogh
Animals 2023, 13(8), 1421; https://doi.org/10.3390/ani13081421 - 21 Apr 2023
Cited by 4 | Viewed by 2354
Abstract
Our objective was to determine a clinically relevant range of centrifugation parameters for processing canine semen. We hypothesized that higher gravitational (g) force and longer time of centrifugation would result in improved spermatozoa recovery rate (RR) but poorer semen quality. Cooled storage under [...] Read more.
Our objective was to determine a clinically relevant range of centrifugation parameters for processing canine semen. We hypothesized that higher gravitational (g) force and longer time of centrifugation would result in improved spermatozoa recovery rate (RR) but poorer semen quality. Cooled storage under standard shipping conditions was used as a stressor to evaluate long-term treatment effects. Individual ejaculates collected from 14 healthy dogs were split into six treatment groups (400 g, 720 g, and 900 g for 5 or 10 min). Sperm RR (%) was calculated post-centrifugation, and plasma membrane integrity (%, Nucleocounter® SP-100™), total and progressive motility (%, subjective and computer-assisted sperm analysis), and morphology (%, eosin-nigrosin staining) were assessed on initial raw semen (T0), post-centrifugation (T1), and 24 h (T2) and 48 h (T3) after cooling. Sperm losses were minimal, and RRs were similar across treatment groups (median >98%, p ≥ 0.062). Spermatozoa membrane integrity was not different between centrifugation groups at any time point (p ≥ 0.38) but declined significantly during cooling (T1 vs. T2/T3, p ≤ 0.001). Similarly, total and progressive motility did not differ across treatments but declined in all groups from T1 to T3 (p ≤ 0.02). In conclusion, our study showed that centrifugation within a range of 400 g–900 g for 5–10 min is appropriate for processing canine semen. Full article
(This article belongs to the Section Animal Reproduction)
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10 pages, 1149 KiB  
Article
Using Dextran Instead of Egg Yolk in Extender for Cryopreservation of Spermatozoa of Dogs of Different Ages
by Taisiia Yurchuk, Olena Pavlovich and Maryna Petrushko
Animals 2022, 12(24), 3480; https://doi.org/10.3390/ani12243480 - 9 Dec 2022
Cited by 3 | Viewed by 2316
Abstract
Egg yolk is a very common supplement of extenders aimed to protect sperm from cryoinjury, but due to their biological risks and difficulties with media standardization, there is a search for alternative. In addition, sperm cryoresistance can be affected by the initial decrease [...] Read more.
Egg yolk is a very common supplement of extenders aimed to protect sperm from cryoinjury, but due to their biological risks and difficulties with media standardization, there is a search for alternative. In addition, sperm cryoresistance can be affected by the initial decrease of their functional characteristics caused by age. The aim of this work was to evaluate the efficiency of using dextran (molecular weight 500 kDa) in the extenders instead of egg yolk for the cryopreservation of spermatozoa of dogs (Chinese Crested breed) of different ages. The obtained ejaculates were divided into three groups depending on the animal’s age: 1–3, 4–6 and 7–10 years old. Sperm was cryopreserved by using 7% glycerol and 20% egg yolk, or 20% dextran. The cryoresistance of spermatozoa of the oldest age category was dramatically decreased, which was manifested in their morphology, motility, and DNA fragmentation rate. There were no differences between the cryoprotectant effect of the dextran-based extender on spermatozoa and the egg yolk-based extender in all age categories of dogs. However, given the benefits of dextran-containing media, its use for the cryopreservation of canine spermatozoa has potential benefits that need to be confirmed by sperm fertilization outcomes. Full article
(This article belongs to the Special Issue Advances in Animal Fertility Preservation)
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23 pages, 3103 KiB  
Article
Age-Dependent Variations in Functional Quality and Proteomic Characteristics of Canine (Canis lupus familiaris) Epididymal Spermatozoa
by Anna Zmudzinska, Jerzy Wisniewski, Piotr Mlynarz, Beata Olejnik and Marzena Mogielnicka-Brzozowska
Int. J. Mol. Sci. 2022, 23(16), 9143; https://doi.org/10.3390/ijms23169143 - 15 Aug 2022
Cited by 9 | Viewed by 2823
Abstract
Increased male age is associated with a significant reduction in semen quality. Little is known about the sperm proteome changes resulting from the aging process. This study aimed to investigate the relationship between the functional quality and proteome of epididymal spermatozoa of dogs [...] Read more.
Increased male age is associated with a significant reduction in semen quality. Little is known about the sperm proteome changes resulting from the aging process. This study aimed to investigate the relationship between the functional quality and proteome of epididymal spermatozoa of dogs that were differing in age. The study was conducted on 30 male dogs that were divided into three age groups. G1—12 to 41 months old, G2—42 to 77 months old, and G3—78 to 132 months old. The sperm samples were assessed using a computer-assisted semen analysis (CASA). The epididymal sperm proteins were analyzed using gel electrophoresis (SDS-PAGE), nano-liquid chromatography coupled to quadrupole time of flight mass spectrometry (NanoUPLC-Q-TOF/MS) and bioinformatic tools. The sperm quality parameters were significantly lower in older dogs. NanoUPLC-Q-TOF/MS identification resulted in 865 proteins that were found in the G1, 472 in G2, and 435 in G3. There were seven proteins that were present in all three age groups, and four of them (ACTB, CE10, NPC2, CRISP2) showed significant changes among the studied groups. Age-dependent variations were detected in the sperm proteome composition and were related to important metabolite pathways, which might suggest that several proteins are implicated in sperm maturation and could be potential aging biomarkers. Full article
(This article belongs to the Special Issue Omics Strategies in Male Reproduction)
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12 pages, 897 KiB  
Article
Effect of Aqueous Extract of Maca Addition to an Extender for Chilled Canine Semen
by Chiara Del Prete, Alfonso Calabria, Valentina Longobardi, Veronica Palumbo, Barbara Merlo, Eleonora Iacono, Simona Tafuri, Domenico Carotenuto, Francesca Ciani, Sara Damiano, Roberto Ciarcia and Natascia Cocchia
Animals 2022, 12(13), 1638; https://doi.org/10.3390/ani12131638 - 26 Jun 2022
Cited by 6 | Viewed by 2816
Abstract
Antioxidant supplementation has been proposed as a new strategy to improve the long-term preservation of semen. The aim of this study was to evaluate the effect of Maca supplementation of semen extender on quality-related canine semen parameters during cooling. Ejaculates from nine dogs [...] Read more.
Antioxidant supplementation has been proposed as a new strategy to improve the long-term preservation of semen. The aim of this study was to evaluate the effect of Maca supplementation of semen extender on quality-related canine semen parameters during cooling. Ejaculates from nine dogs were cooled for 7 days in the absence (control group) or in the presence of 10, 20 and 50 μL/mL of an aqueous extract of Maca. Sperm were evaluated for sperm viability, motility, DNA fragmentation and lipid peroxidation after 3 h, 24 h, 4 days and 7 days of storage. The addition of 10 μL/mL of Maca preserved sperm DNA and plasma membrane integrity at 3 h and increased sperm curvilinear velocity after 24 h. Treatment with 20 and 50 μL/mL of Maca increased the percentage of hyperactivated sperm after 3 h. Moreover, semen treated with 20 μL/mL of Maca decreased lipid peroxidation at 24 h. A significant reduction of sperm DNA and plasma membrane integrity as well as of kinetics parameters between 3 and 24 h of refrigerated storage with the higher concentration tested was observed. Although Maca was not able to protect canine semen with extended refrigeration storage time, it increased hyperactivation and preserved DNA integrity in short-term storage. Full article
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12 pages, 1797 KiB  
Article
A Comparative Study of the Effects of Osaterone Acetate and Deslorelin Acetate on Sperm Kinematics and Morpho-Functional Parameters in Dogs
by Wojciech Niżański, Maria Eberhardt, Małgorzata Ochota, Christelle Fontaine, Xavier Levy and Joanna Pasikowska
Animals 2022, 12(12), 1548; https://doi.org/10.3390/ani12121548 - 15 Jun 2022
Cited by 7 | Viewed by 2572
Abstract
Benign prostatic hyperplasia (BPH) is one of the most common problems in older male dogs that often has a huge impact on their health and welfare. This article presents a comparison between osaterone acetate (Ypozane®; Virbac®)(OA) and deslorelin acetate [...] Read more.
Benign prostatic hyperplasia (BPH) is one of the most common problems in older male dogs that often has a huge impact on their health and welfare. This article presents a comparison between osaterone acetate (Ypozane®; Virbac®)(OA) and deslorelin acetate (Suprelorin®; Virbac®)(DA), medications that are the main therapeutic alternative to castration in dogs with BPH. Forty dogs were divided into four groups: I—negative control (five dogs without BPH); II—positive control (10 individuals diagnosed with BPH); III—15 dogs treated with DA, and IV—10 individuals treated with OA. Semen fractions were collected on days 0 (day of treatment), 7, 14, and 21, and weeks 8, 12, 16, and 20. Macroscopic, microscopic and CASA analyses were performed. Both DA and OA significantly affected the properties of the canine ejaculate. The DA lead to the lack of libido and had lesser effects to the sperm function before it caused azoospermia, whereas OA had no effect on libido and only temporary reduction in seminal plasma volume was observed, which resulted in temporary deterioration in the percentage of motile and progressive spermatozoa. Full article
(This article belongs to the Section Animal Reproduction)
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21 pages, 2945 KiB  
Article
Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine (Canis lupus familiaris) Epididymal Spermatozoa and Sperm Structure Separation
by Anna Zmudzinska, Mariusz A. Bromke, Rafal Strzezek, Magdalena Zielinska, Beata Olejnik and Marzena Mogielnicka-Brzozowska
Animals 2022, 12(6), 772; https://doi.org/10.3390/ani12060772 - 18 Mar 2022
Cited by 8 | Viewed by 3369
Abstract
This study was provided for proteomic analysis of intracellular and membrane-associated fractions of canine (Canis lupus familiaris) epididymal spermatozoa and additionally to find optimal sonication parameters for the epididymal sperm morphological structure separation and sperm protein isolation. Sperm samples were collected [...] Read more.
This study was provided for proteomic analysis of intracellular and membrane-associated fractions of canine (Canis lupus familiaris) epididymal spermatozoa and additionally to find optimal sonication parameters for the epididymal sperm morphological structure separation and sperm protein isolation. Sperm samples were collected from 15 dogs. Sperm protein fractions: intracellular (SIPs) and membrane-associated (SMAPs) were isolated. After sonication, sperm morphology was evaluated using Spermac Stain™. The sperm protein fractions were analyzed using gel electrophoresis (SDS-PAGE) and nanoliquid chromatography coupled to quadrupole time-of-flight mass spectrometry (NanoLC-Q-TOF/MS). UniProt database-supported identification resulted in 42 proteins identified in the SIPs and 153 proteins in the SMAPs. Differentially abundant proteins (DAPs) were found in SIPs and SMAPs. Based on a gene ontology analysis, the dominant molecular functions of SIPs were catalytic activity (50%) and binding (28%). Hydrolase activity (33%) and transferase activity (21%) functions were dominant for SMAPs. Bioinformatic analysis of SIPs and SMAPs showed their participation in important metabolic pathways in epididymal sperm, which may suggest their potential as sperm quality biomarkers. The use of sonication 150 W, 10 min, may be recommended for the separation of dog epididymal sperm heads, tails, acrosomes and the protein isolation. Full article
(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)
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12 pages, 721 KiB  
Review
Canine Spermatozoa—Predictability of Cryotolerance
by Sabine Schäfer-Somi, Martina Colombo and Gaia Cecilia Luvoni
Animals 2022, 12(6), 733; https://doi.org/10.3390/ani12060733 - 15 Mar 2022
Cited by 7 | Viewed by 3557
Abstract
Markers of freezability allow the selection of ejaculates of good freezability. So far, most investigations were conducted in boars, bulls, rams and horses, with high economic interests triggering the efforts. The progress in dogs is comparably slow. A critical evaluation of the methods [...] Read more.
Markers of freezability allow the selection of ejaculates of good freezability. So far, most investigations were conducted in boars, bulls, rams and horses, with high economic interests triggering the efforts. The progress in dogs is comparably slow. A critical evaluation of the methods requires consideration of practicability, with most labs not even possessing a computer assisted sperm analyser (CASA); furthermore, small canine ejaculates mostly do not allow the use of large semen volumes. In dogs, modern markers of freezability no longer assess single membrane constituents or seminal plasma components but comprise tests of cell functionality and adaptability, energy metabolism, cluster analyses of kinetic and morphometric parameters, as well as DNA intactness. Identification of the most efficient combination of tests seems useful. At present, examination by CASA combined with cluster analysis of kinetic subgroups, JC-1 staining and COMET assay or staining with toluidine blue seem most appropriate; however, cell volumetry and other functional tests deserve better attention. A better understanding of spermatozoa energy metabolism might reveal new markers. This review focuses on the requirements and markers of freezability of canine semen, highlighting potential future candidates. Full article
(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)
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9 pages, 1156 KiB  
Article
Sperm Gone Smart: A Portable Device (iSperm®) to Assess Semen Concentration and Motility in Dogs
by Guillaume Domain, Penelope Banchi, Hiba Ali Hassan, Anouk Eilers, Joke Lannoo, Eline Wydooghe, Wojciech Niżański and Ann Van Soom
Animals 2022, 12(5), 652; https://doi.org/10.3390/ani12050652 - 4 Mar 2022
Cited by 6 | Viewed by 4130
Abstract
The iSperm® is a portable device for semen analysis. This study aimed to investigate its correlation with a conventional computer-assisted sperm analyzer (ISAS®v1) for the assessment of semen concentration and kinematic parameters in dogs (n = 224). The intra-assay [...] Read more.
The iSperm® is a portable device for semen analysis. This study aimed to investigate its correlation with a conventional computer-assisted sperm analyzer (ISAS®v1) for the assessment of semen concentration and kinematic parameters in dogs (n = 224). The intra-assay variability of both devices and their ability to estimate semen concentration at a fixed value of 40 × 106/mL were also investigated. Results showed that the intra-assay variability was lower for the ISAS®v1 for all parameters compared to the iSperm®. Hence, iSperm® estimates were more variable in-between fields. Both the iSperm® and the ISAS®v1 were not reliable in estimating semen concentration (ISAS®v1: median 30 × 106/mL, interquartile range (IQR) 12, p < 0.01; iSperm®: median 35.12 × 106/mL, IQR 11.11, p < 0.01). Finally, positive correlations were found between both devices with stronger correlations obtained when four fields were analyzed by the iSperm®. However, the low number of spermatozoa analyzed per field and the inability to avoid artifacts are downsides that currently limit the reliability of the iSperm®. Therefore, the software of iSperm® needs some improvement to make it a valid and practical alternative to automated computerized systems for the analysis of canine semen. Full article
(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)
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11 pages, 413 KiB  
Article
The Use of κ-Carrageenan in Egg Yolk Free Extender Improves the Efficiency of Canine Semen Cryopreservation
by Eunji Kim, Areeg Almubarak, Nabeel Talha, Il-Jeoung Yu and Yubyeol Jeon
Animals 2022, 12(1), 88; https://doi.org/10.3390/ani12010088 - 31 Dec 2021
Cited by 2 | Viewed by 2511
Abstract
κ-Carrageenan is a plant polysaccharide derived from red seaweeds reported to possess potential medicinal and antioxidants activities. The present study aimed to identify the cryoprotective effects of κ-carrageenan on the quality of frozen-thawed canine semen. Twenty-eight ejaculates were collected and diluted in a [...] Read more.
κ-Carrageenan is a plant polysaccharide derived from red seaweeds reported to possess potential medicinal and antioxidants activities. The present study aimed to identify the cryoprotective effects of κ-carrageenan on the quality of frozen-thawed canine semen. Twenty-eight ejaculates were collected and diluted in a Tris egg-yolk-free extender supplemented with various concentrations of κ-carrageenan (0.0%, 0.1%, 0.2%, 0.3%, and 0.5%). The addition of κ-carrageenan to the extender at a 0.2% concentration induced a significant increase in the total motility (TM) and the rapid progressive motility (RPM) of canine sperm. Among the experimental groups, the highest percentage of sperms with intact acrosomes was found in the 0.5% κ-carrageenan group (p < 0.05). Apoptosis levels were significantly lower in the 0.1% and 0.2% κ-carrageenan treatment. Moreover, sperm in the κ-carrageenan supplemented group showed a significantly higher expression of antiapoptotic (Bcl-2) and lower expression of NADPH oxidase (NOX5), spermine synthase (SMS), and spermine oxidase (SMOX) genes than those in the control group. In conclusion, the addition of κ-carrageenan to the freezing extender improved the overall efficiency of frozen-thawed dog spermatozoa. Full article
(This article belongs to the Section Animal Reproduction)
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