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4 pages, 202 KiB

Open AccessEditorial

Current Status and Advances in Semen Preservation

by Anna Dziekońska and Agnieszka Partyka

Animals 2023, 13(1), 123; https://doi.org/10.3390/ani13010123 - 28 Dec 2022

Cited by 1 | Viewed by 1262

Abstract

Recent advances in assisted reproductive technology (ART) have increased the effectiveness of fertility treatments [...] Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

Research

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14 pages, 2583 KiB

Open AccessArticle

Fluorescence Microscopy and Flow-Cytometry Assessment of Substructures in European Red Deer Epididymal Spermatozoa after Cryopreservation

by Anna Dziekońska, Marek Lecewicz, Agnieszka Partyka and Wojciech Niżański

Animals 2023, 13(6), 990; https://doi.org/10.3390/ani13060990 - 08 Mar 2023

Cited by 1 | Viewed by 1829

Abstract

Thawed spermatozoa, sampled post mortem from the fresh epididymides of European red deer and epididymides stored for up to 12 h at 2–4 °C, were evaluated by fluorescence microscopy (FM) and flow cytometry (FC). The sperm samples were extended and cryopreserved. The sperm [...] Read more.

Thawed spermatozoa, sampled post mortem from the fresh epididymides of European red deer and epididymides stored for up to 12 h at 2–4 °C, were evaluated by fluorescence microscopy (FM) and flow cytometry (FC). The sperm samples were extended and cryopreserved. The sperm motility (CASA), sperm viability (SYBR⁺/PI^-), acrosome integrity, mitochondrial activity, apoptotic changes, and chromatin stability were assessed. Sperm were analyzed by FM before cryopreservation, and by FM and FC after thawing. Epididymal storage time (for 12 h) had no significant effect (p > 0.05) on the examined variables before cryopreservation. After thawing, the storage variants differed (p ˂ 0.05) in the percentage of apoptotic sperm (FM and FC) and DNA integrity (FC). The results of FM and FC differed (p ˂ 0.05) in all the analyzed parameters, excluding SYBR⁺/PI. Significant correlations (p ˂ 0.01) were observed between the sperm viability, acrosome integrity, and the percentage of non-apoptotic spermatozoa, regardless of the applied technique. In FM, the above parameters were also significantly correlated with mitochondrial activity. The study demonstrated that European red deer spermatozoa stored in the epididymides at 2–4 °C for 12 h can be used for cryopreservation. Both techniques were equally reliable, but FM was better suited for evaluating mitochondrial activity whereas FC was more useful in the evaluation of DNA fragmentation. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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9 pages, 973 KiB

Open AccessArticle

Comparison of Commercial Poultry Semen Extenders Modified for Cryopreservation Procedure in the Genetic Resource Program of Czech Golden Spotted Hen

by Kristýna Petričáková, Martina Janošíková, Martin Ptáček, Lukáš Zita, Filipp Georgijevič Savvulidi and Agnieszka Partyka

Animals 2022, 12(20), 2886; https://doi.org/10.3390/ani12202886 - 21 Oct 2022

Cited by 4 | Viewed by 1780

Abstract

Spermatozoa cryoconservation represents an important strategy for partial in vitro or rescue programs designed for threatened livestock populations. The procedure for the semen cryopreservation of the Czech Golden Spotted Hen was proposed due to the lower fertilization rate of poultry semen compared to [...] Read more.

Spermatozoa cryoconservation represents an important strategy for partial in vitro or rescue programs designed for threatened livestock populations. The procedure for the semen cryopreservation of the Czech Golden Spotted Hen was proposed due to the lower fertilization rate of poultry semen compared to mammalian species. The aim of this study was to compare commercial extenders designed for liquid storage preservation with the use of a predefined cryoprotectant, and, thus, to propose an important tool for the procedure of the semen cryopreservation of the Czech Golden Spotted Hen. Ejaculates were sampled from four roosters during five semen collection days. The samples were frozen in Poultry media^®, Raptac^® and NeXcell^® extenders supplemented with a 9% N-methylacetamide (NMA) cryoprotectant. Sperm parameters of the total motility (MOT; %), plasma membrane and acrosome intactness (PAI; %), plasma membrane damage (%), acrosome damage (%) and cells with plasma membrane and acrosome damage (%) were assessed using a mobile mCASA analyzer and flow cytometer after the cryopreservation of the insemination doses (IDs). For Poultry media^® (PAI = 51.11%; MOT = 23.58%) and Raptac^® (PAI = 52.04%; MOT = 23.13%) extenders with the addition of an NMA cryoprotectant, the comparable results were detected after thawing. For NexCell^® media, the results were poor (PAI = 7.07%; MOT = 3.83%). Our results indicated two extenders suitable for the cryopreservation procedure, with the applied modification. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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13 pages, 455 KiB

Open AccessArticle

Effect of Dimethylacetamide Concentration on Motility, Quality, Antioxidant Biomarkers, Anti-Freeze Gene Expression, and Fertilizing Ability of Frozen/Thawed Rooster Sperm

by Gamal M. K. Mehaisen, Ahmed M. Elomda, Shaimaa K. Hamad, Mona M. Ghaly, Yanyan Sun, Yunlei Li, Yunhe Zong, Jilan Chen, Agnieszka Partyka, Ali Nazmi, Ahmed O. Abbas and Farid K. R. Stino

Animals 2022, 12(20), 2739; https://doi.org/10.3390/ani12202739 - 12 Oct 2022

Cited by 11 | Viewed by 1645

Abstract

Sperm cryopreservation is of great importance for the poultry industry but still needs to be optimized. The high susceptibility of poultry sperm to cryodamage leads to low fertility rates after cryopreservation. Therefore, the present study aimed at evaluating the effect of including a [...] Read more.

Sperm cryopreservation is of great importance for the poultry industry but still needs to be optimized. The high susceptibility of poultry sperm to cryodamage leads to low fertility rates after cryopreservation. Therefore, the present study aimed at evaluating the effect of including a cryoprotectant, dimethylacetamide (DMA), in the chicken semen freezing extenders at a final concentration of 3%, 6%, or 9% on the post-thawed sperm motility, quality, antioxidant biomarkers, anti-freeze gene expression, and fertilizing ability. Results showed that the total motile sperm, progressivity, and viability were quadratically increased (p < 0.05) in the 6% DMA group. The antioxidant enzyme activity and lipid peroxidation were negatively (p < 0.05) affected by the increase in DMA concentration. Furthermore, some anti-freeze-associated genes such as heat shock protein 70 (HSP70) and ras homolog family member A (RHOA) were linearly and quadratically down-regulated (p < 0.05) with the high concentration of DMA. Finally, the fertility and hatchability rates did not indicate statistical differences between DMA groups. It can be concluded that using the low concentration of 3–6% DMA in the freezing semen extender is preferable to obtain acceptable results in the post-thawed sperm quality and fertility. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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14 pages, 1491 KiB

Open AccessArticle

The Effect of Different Extenders on the Quality Characteristics of European Red Deer Epididymal Sperm Stored at 5 °C

by Anna Dziekońska, Nicoletta M. Neuman, Klaudia K. Burdal, Agnieszka Wiszniewska-Łaszczych and Marek Bogdaszewski

Animals 2022, 12(19), 2669; https://doi.org/10.3390/ani12192669 - 04 Oct 2022

Cited by 4 | Viewed by 1575

Abstract

The aim of this study was to evaluate the effect of different extenders on the quality of European red deer epididymal sperm stored at 5 °C. Epididymal spermatozoa were collected post mortem from 10 stags and diluted with three extenders (Bovidyl^®, [...] Read more.

The aim of this study was to evaluate the effect of different extenders on the quality of European red deer epididymal sperm stored at 5 °C. Epididymal spermatozoa were collected post mortem from 10 stags and diluted with three extenders (Bovidyl^®, BoviFree^®, and BioXcell^®) and stored at 5 °C. Sperm motility (TMOT), motility parameters (system CASA), plasma membrane integrity (SYBR-14⁺/PI⁻), acrosomal membrane integrity (FITC-PNA⁻/PI⁻), mitochondrial activity (JC-1/PI), viability, and apoptotic-like changes (YOPRO/PI) were evaluated. The analyses were conducted on the first and successive days of storage (D1–D7). The applied extender, storage time, and the interactions between these factors significantly (p < 0.001) affected most of the analyzed parameters whose values were highest in sperm samples stored in Bovidyl^®, regardless of storage time. In Bovidyl^®, BoviFree^®, and BioXcell^® extenders, TMOT values were estimated at 83%, 63%, and 59%, respectively, on D3. The extenders significantly influenced DNA integrity on D7. The percentage of dead sperm increased from D3. The quality of stored sperm cells was significantly influenced by the extenders’ biochemical composition. BoviFree^® and BioXcell^® contain glycerol which could contribute to deteriorating the quality of spermatozoa stored at 5 °C. Sperm cells stored in the egg yolk-based extender (Bovidyl^®) were characterized by the highest viability and functionality. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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13 pages, 1833 KiB

Open AccessArticle

Beneficial Effect of Proline Supplementation on Goat Spermatozoa Quality during Cryopreservation

by Weijing Zhang, Lingjiang Min, Yajing Li, Yaning Lang, S. A. Masudul Hoque, Adedeji Olufemi Adetunji and Zhendong Zhu

Animals 2022, 12(19), 2626; https://doi.org/10.3390/ani12192626 - 30 Sep 2022

Cited by 13 | Viewed by 1870

Abstract

Sperm cryopreservation contributes to the extensive utilization of artificial insemination (AI) in the daily livestock industry. However, due to the presence of few sperm with good biological function in post-thaw goat sperm, its use has been limited for AI purposes. Hence, its improvement [...] Read more.

Sperm cryopreservation contributes to the extensive utilization of artificial insemination (AI) in the daily livestock industry. However, due to the presence of few sperm with good biological function in post-thaw goat sperm, its use has been limited for AI purposes. Hence, its improvement has been the focus of many research studies. This study aimed to investigate the effects of proline supplementation of the freezing medium on goat sperm. The goat semen was cryopreserved with freezing medium supplementation of different concentrations of proline (0, 0.5, 1, 2 and 4 mM). The post-thaw sperm motility patterns, membrane integrity, acrosome integrity, lipid peroxidation (LPO) levels, malondialdehyde (MDA) levels, total antioxidant capacity (T-AOC), proline dehydrogenase (PRODH) activity, superoxide dis-mutase (SOD) activity, glutathione (GSH) levels and GSH/GSSG were evaluated. Likewise, the expression and immunofluorescent localization of PRODH in post-thaw goat sperm was also detected. It was observed that addition of 2 mM proline to the freezing medium significantly enhanced post-thaw goat sperm total motility, progressive motility, straight-linear velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), straightness (STR), linearity (LIN), membrane integrity and acrosome integrity. Interestingly, PRODH was expressed in post-thaw goat sperm, especially in the post-acrosome and sperm tail. Addition of 2 mM proline also significantly increased the post-thaw sperm PRODH activity compared to the control. Moreover, post-thaw goat sperm LPO levels and MDA levels were reduced by supplementation of 2 mM proline. Furthermore, compared to the control, the values of post-thaw goat sperm T-AOC, SOD activity, GSH level and GSH/GSSG were also significantly increased in 2 mM proline treatment. Reduction of post-thaw goat sperm apoptosis in 2 mM proline treatment was also observed as the levels of Caspase3 and Caspase9 were decreased by the supplementation with 2 mM proline. These observations suggest that the addition of 2 mM proline to the freezing medium increased post-thaw goat sperm quality by reducing oxidative stress during cryopreservation. These findings also provide novel insights into the use of proline as an efficient additive to enhance post-thaw goat sperm quality during cryopreservation. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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12 pages, 556 KiB

Open AccessArticle

Morphometry of Boar Spermatozoa in Semen Stored at 17 °C—The Influence of the Staining Technique

by Dorota Szablicka, Anna Wysokińska, Angelika Pawlak and Klaudia Roman

Animals 2022, 12(15), 1888; https://doi.org/10.3390/ani12151888 - 25 Jul 2022

Cited by 2 | Viewed by 2313

Abstract

The aim of the study was to assess the morphometry of sperm during storage of liquid boar semen at 17 °C. An attempt was also made to evaluate the suitability of three staining methods for assessment of boar sperm morphometry. The study was [...] Read more.

The aim of the study was to assess the morphometry of sperm during storage of liquid boar semen at 17 °C. An attempt was also made to evaluate the suitability of three staining methods for assessment of boar sperm morphometry. The study was carried out on 20 Landrace boars. Semen was collected from the boars every 5 days by the manual method. Four ejaculates from each boar were analysed (80 ejaculates in total). Analyses were performed five times: at 1 h, 24 h, 48 h, 96 h, and 168 h after semen collection. Blisters with insemination doses were opened immediately before the analyses. From each insemination dose, smears were prepared for morphometric evaluation of sperm, which were stained by three methods (eosin-nigrosin—EN, eosin-gentian—EG, and SpermBlue—SB). Morphometric measurements of 15 randomly selected sperm with normal morphology were performed on each slide. The morphometric measurements included the following parameters: sperm head length, width, area, and perimeter; tail length; and total sperm length. The results of the morphometric measurements were used to calculate the head shape index. The morphometric dimensions of the sperm were shown to change during storage of semen at 17 °C. The extent of these changes, however, depended on the staining method used, as the three methods result in different morphometric dimensions of sperm, in the case of both the head and the tail. In the slides stained by the eosin-nigrosin method, the dimensions of the head and tail were smaller at every time of storage than in the slides stained by the SpermBlue and eosin-gentian methods. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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14 pages, 1230 KiB

Open AccessArticle

Cryopreservation of Sperm from an Endangered Snake with Tests of Post-Thaw Incubation in Caffeine

by Mark R. Sandfoss, Jessica Cantrell, Beth M. Roberts and Steve Reichling

Animals 2022, 12(14), 1824; https://doi.org/10.3390/ani12141824 - 17 Jul 2022

Cited by 3 | Viewed by 1875

Abstract

Cryopreservation of sperm to preserve the genetic diversity of declining populations is a promising technique to aid in the recovery of endangered species such as the Louisiana pinesnake (Pituophis ruthveni). However, this technique has been performed on only a handful of [...] Read more.

Cryopreservation of sperm to preserve the genetic diversity of declining populations is a promising technique to aid in the recovery of endangered species such as the Louisiana pinesnake (Pituophis ruthveni). However, this technique has been performed on only a handful of snake species and with limited success. Here, we tested a cryoprotective agent (CPA) mixture containing Lake’s buffer with 10% N,N-dimethyl formamide (DMF), 2% methanol, 5% clarified egg yolk, (v/v% final concentration) against 16 other CPA-treatment mixtures. These contained either Lake’s buffer or TEST egg yolk buffer as the base diluent with a penetrating or non-penetrating CPA on the post-thaw recovery of sperm motility and viability. We also investigated the effect of post-thaw incubation treatment in TL HEPES supplemented with 10% fetal bovine serum (H10) alone or with caffeine on post-thaw motility parameters. Sperm from 16 Louisiana pinesnakes was cryopreserved, and the effectiveness of the CPA treatment mixtures and post-thaw treatments was determined based on measurements of sperm motility and viability. Sperm cryopreservation significantly reduced initial post-thaw sperm quality for all of the extender treatments. Viability of sperm was best maintained when cryopreserved in an CPA treatment mixture containing Lake’s buffer with 10% DMF, 2% methanol, and 5% clarified egg yolk with the addition of 5 mg/mL bovine serum albumin (BSA). For several extender mixtures a similar percent of post-thaw motility was observed, but no forward motility returned in any post-thaw samples prior to incubation in dilution treatments. Following incubation in both post-thaw treatments, the percent of forward motility and the index of forward progressive movement improved significantly. Post-thaw dilution with H10 containing caffeine improved motility parameters over H10 alone, suggesting further investigation of post-thaw treatment in caffeine could be beneficial. Although, cryopreservation of sperm from the Louisiana pinesnake continues to present a challenge, post-thaw dilution and the addition of BSA to CPA mixtures provides areas for improving cryopreservation methods for this endangered species. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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21 pages, 2945 KiB

Open AccessArticle

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 4 | Viewed by 2334

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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13 pages, 302 KiB

Open AccessArticle

Spermatozoa Survival in Egg Yolk-Based and Soybean-Based Extenders at Ambient and Chilling Temperature in Domestic Turkeys (Meleagris gallopavo)

by Isa Mohammed Alkali, Suleiman Omeiza Asuku, Martina Colombo, Muhammad Modu Bukar, Mohammed Ahmed Waziri and Gaia Cecilia Luvoni

Animals 2022, 12(5), 648; https://doi.org/10.3390/ani12050648 - 03 Mar 2022

Cited by 2 | Viewed by 1720

Abstract

Populations of many galliform species have declined mainly due to habitat loss and over-hunting, notably the Congo peacock, which has been classified as a vulnerable species by the International Union for Conservation of Nature (IUCN). The domestic turkey, being a species of least [...] Read more.

Populations of many galliform species have declined mainly due to habitat loss and over-hunting, notably the Congo peacock, which has been classified as a vulnerable species by the International Union for Conservation of Nature (IUCN). The domestic turkey, being a species of least concern, which has been reported to be closely related to peacocks, could serve as a model for the optimization of assisted reproductive technologies for the Congo peacock. This study was aimed at developing a suitable turkey semen extender for artificial insemination in field conditions. Semen was collected using the dorso-abdominal massage technique from seven turkey toms and analyzed. Ejaculates with >70% motility and >80% live spermatozoa were pooled and divided into four aliquots (four treatments). Each of the four treatments was extended in a soybean-based extender or an egg yolk-based extender, with or without L-ascorbic acid. Two liquid preservation protocols (ambient temperature (35 °C) and chilled (4 °C)) were employed, and quality parameters including motility, viability and morphology were evaluated. The results show that the two extenders were similar with regard to semen quality parameters, and L-ascorbic acid supplementation of the turkey semen extenders improved semen quality during liquid storage. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

16 pages, 2542 KiB

Open AccessArticle

Influence of Zinc and Manganese Nanoparticles on Selected Parameters of Turkey Spermatozoa Stored in a Liquid State at 4 °C

by Aleksandra Orzołek, Katarzyna T. Rafalska, Wiktoria A. Otowska, Władysław Kordan, Anna J. Korzekwa and Krzysztof Kozłowski

Animals 2021, 11(11), 3289; https://doi.org/10.3390/ani11113289 - 17 Nov 2021

Cited by 11 | Viewed by 1980

Abstract

The aim of this study was to determine the effect of semen extender supplementation with 25 or 50 μM of zinc nanoparticles (ZnNPs) or manganese nanoparticles (MnNPs) on turkey spermatozoa preserved in a liquid state. Twenty turkey ejaculates were obtained from twenty healthy [...] Read more.

The aim of this study was to determine the effect of semen extender supplementation with 25 or 50 μM of zinc nanoparticles (ZnNPs) or manganese nanoparticles (MnNPs) on turkey spermatozoa preserved in a liquid state. Twenty turkey ejaculates were obtained from twenty healthy males. The collected semen was preserved at 4 °C for 48 h with or without NPs. Selected qualitative and quantitative parameters of sperm (motility, plasma membrane activity, mitochondrial membrane potential (MMP) and the percentage of sperm demonstrating NO and SOD activity) were examined after 2, 24 and 48 h of storage. Sperm motility and MMP decreased in semen preserved with ZnNPs at each time point of the analysis. However, all spermatozoa remained viable throughout storage. In contrast, membrane integrity and mitochondria activity (p ≤ 0.05) increased, and the highest SOD activity (p ≤ 0.05) was observed in semen preserved with MnNPs. The addition of MnNPs to the semen extender could potentially improve the parameters of turkey semen during prolonged storage. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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15 pages, 436 KiB

Open AccessArticle

From the Semen Collection Method to the Hatchlings: The Use of Cryopreserved Sperm from Pheasants Fed an Antioxidant-Enriched Diet

by Annelisse Castillo, Carla Lenzi, Andrea Pirone, Alessandro Baglini, Claudia Russo, Dominga Soglia, Achille Schiavone and Margherita Marzoni Fecia di Cossato

Animals 2021, 11(9), 2624; https://doi.org/10.3390/ani11092624 - 07 Sep 2021

Cited by 7 | Viewed by 2824

Abstract

A widely used approach to preserving genetic diversity in birds involves the cryopreservation of semen. In this process, cells are subjected to physical and chemical stresses, but not all cell species respond equally. Many studies have been published on the freezing–thawing of sperm [...] Read more.

A widely used approach to preserving genetic diversity in birds involves the cryopreservation of semen. In this process, cells are subjected to physical and chemical stresses, but not all cell species respond equally. Many studies have been published on the freezing–thawing of sperm cells from a wide variety of domestic and wild species, on issues ranging from the sperm quality to different protocols, fertilisation success rates, etc. Nevertheless, very little information is available on the common pheasant. To fill this gap, the aim of this study was to describe the pheasant semen collection method, evaluate some qualitative parameters of sperm from males fed an antioxidant-enriched diet, and to test the in vivo fertilising capacity of the cryo-preserved semen. The freezing protocol employed involved pellets thawed by the hotplate method. Dimethylacetamide was used as a cryoprotectant at a final concentration of 6%. A total of six AIs were performed at 3-4-day intervals on a total of 40 females with doses of 35 × 10⁶ of normal live thawed sperm. Males receiving the enriched diet produce more abundant and concentrated ejaculates. Freeze–thawed sperm lost 85% of their initial mobility, and diet influenced neither sperm mobility nor viability. The enriched diet did improve the number of normal freeze–thawed cells and was associated with a lower sperm fracture incidence. Regardless of the dietary group, frozen–thawed sperm resulted in a fertility rate of 30%, with 8-9 chicks hatching for every 100 eggs incubated. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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12 pages, 2085 KiB

Open AccessArticle

A High Incidence of Sperm with Cytoplasmic Droplets Affects the Response to Bicarbonate in Preserved Boar Semen

by Heiko Henning, Anne-Marie Luther and Dagmar Waberski

Animals 2021, 11(9), 2570; https://doi.org/10.3390/ani11092570 - 31 Aug 2021

Cited by 4 | Viewed by 3741

Abstract

Retained cytoplasmic droplets (CD) are the most frequent sperm abnormality in boar semen. A high incidence of CD is associated with subfertility, but the underlaying reasons are not well understood. The storage of extended semen might augment the adverse effects of CD on [...] Read more.

Retained cytoplasmic droplets (CD) are the most frequent sperm abnormality in boar semen. A high incidence of CD is associated with subfertility, but the underlaying reasons are not well understood. The storage of extended semen might augment the adverse effects of CD on essential steps towards fertilization, such as capacitation. The aim of this study was to examine whether the enhanced presence of CD in boar semen influences sperm’s response to the capacitation stimulus bicarbonate during long-term semen storage. Extended semen samples (n = 78) from 13 artificial insemination centers were analyzed using a flow cytometric calcium influx assay. Samples with >15% of CD showed a reduced specific response to bicarbonate and a higher non-specific destabilization after storage for 96 h and subsequent incubation at 38 °C in three variants of Tyrode’s medium (p < 0.05). The size of the bicarbonate-responsive sperm population was inversely correlated with the presence of CD-bearing sperm (r = −0.61, p < 0.01). Samples with ≤15% and samples with >15% of CD did not differ in motility or viability and acrosome integrity during semen storage. In conclusion, incomplete epididymal sperm maturation impairs the in vitro capacitation ability and promotes sperm destabilization in stored boar semen. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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13 pages, 1491 KiB

Open AccessArticle

Comparative Proteomic Analysis of Young and Adult Bull (Bos taurus) Cryopreserved Semen

by Błażej Westfalewicz, Mariola Słowińska, Sylwia Judycka, Andrzej Ciereszko and Mariola A. Dietrich

Animals 2021, 11(7), 2013; https://doi.org/10.3390/ani11072013 - 05 Jul 2021

Cited by 10 | Viewed by 4037

Abstract

The age of the bull is widely accepted to influence the production of sperm, affecting the amount and quality of produced semen, which in turn impacts the results of cryopreservation. However, the exact influence of the maturation process on cryopreserved sperm, as well [...] Read more.

The age of the bull is widely accepted to influence the production of sperm, affecting the amount and quality of produced semen, which in turn impacts the results of cryopreservation. However, the exact influence of the maturation process on cryopreserved sperm, as well as the underlying molecular mechanisms of this process, are not fully understood. The goal of this study was to evaluate changes in the proteome of thawed semen (spermatozoa and supernatant) collected from young and adult bulls (n = 6) using the 2D-DIGE approach. The quality of semen was assessed using a CASA system and flow cytometry. We found no significant age-related variation in semen quality, with the exception of the average path velocity of sperm movement, which was higher in adult bulls. Proteomic analysis indicated 15 spermatozoa proteins and 10 supernatant proteins with significant age-related changes. Our results suggest that semen from adult bulls is better equipped with proteins related to energy production, protection of spermatozoa against oxidative stress and fertilizing ability. Proteins increased in abundance in young bull spermatozoa were connected to the cytoskeleton and its development, which strongly suggests that developmental processes are still in progress. In conclusion, our results provide novel insight into the mechanism of the development of the male reproductive system of cattle. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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12 pages, 271 KiB

Open AccessArticle

Beneficial Influence of Soybean Lecithin Nanoparticles on Rooster Frozen–Thawed Semen Quality and Fertility

by Lingwei Sun, Mengqian He, Caifeng Wu, Shushan Zhang, Jianjun Dai and Defu Zhang

Animals 2021, 11(6), 1769; https://doi.org/10.3390/ani11061769 - 13 Jun 2021

Cited by 14 | Viewed by 4516

Abstract

The present study aimed to investigate the impact of different concentrations (0%, 0.5%, 1.0%, 1.5%, and 2.0%) of nano-soybean lecithin (SL) in the extender on sperm quality, sperm motion characteristics, and fertility outcomes of post-thawed rooster semen. Adult Ross broiler breeder roosters (n [...] Read more.

The present study aimed to investigate the impact of different concentrations (0%, 0.5%, 1.0%, 1.5%, and 2.0%) of nano-soybean lecithin (SL) in the extender on sperm quality, sperm motion characteristics, and fertility outcomes of post-thawed rooster semen. Adult Ross broiler breeder roosters (n = 20) were subjected to semen collections twice a week for three weeks. At each collection, semen samples were pooled and allocated into five treatments corresponding to different nano-SL concentrations (control, SL0.5, SL1.0, SL1.5, and SL2.0). Sperm parameters, including motility (collected using a computer-assisted sperm analysis system), plasma membrane and acrosome integrities, and mitochondrial activity were assessed. Sperm malondialdehyde (MDA) and antioxidant activities (total antioxidant capacity (TAC); superoxide dismutase (SOD); glutathione peroxidase (GPx)) were evaluated. The fertility and hatchability obtained with frozen–thawed rooster semen supplemented with the optimum nano-SL concentration were assessed after artificial insemination. The results showed that the addition of 1% nano-SL into the extender led to a higher semen motility in roosters, improved plasma membrane and acrosome integrities, and higher mitochondrial activity of post-thawed rooster semen in comparison to controls (p < 0.05). The MDA levels in the SL0.5 and SL1.0 groups were lower than the other groups (p < 0.05). TAC activities in SL0.5, SL1.0, and SL1.5 groups were significantly higher than those in the other groups (p < 0.05). It was observed that the concentration of SOD was higher in the SL1.0 group than in the other groups (p < 0.05). The activity of GPx was not influenced in any of the cases (p > 0.05). Moreover, the percentages of fertility and hatchability in the SL1.0 group were higher (56.36% and 58.06%) than those in the control group (42.72% and 40.43%). In summary, the addition of nano-SL to the extenders enhanced the post-thawed semen quality and fertility of roosters by reducing the level of oxidative stress. The optimum nano-SL concentration was 1.0%. These results may be beneficial for improving the efficacy of semen cryopreservation procedures in poultry breeding. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

Review

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12 pages, 339 KiB

Open AccessFeature PaperReview

Effect of Sperm Cryopreservation in Farm Animals Using Nanotechnology

by Muhammad Faheem Akhtar, Qingshan Ma, Yan Li, Wenqiong Chai, Zhenwei Zhang, Liangliang Li and Changfa Wang

Animals 2022, 12(17), 2277; https://doi.org/10.3390/ani12172277 - 02 Sep 2022

Cited by 9 | Viewed by 2732

Abstract

Sperm cryopreservation is one of the sublime biotechnologies for assisted reproduction. In recent decades, there has been an increasing trend in the use of preserved semen. Post-thaw semen quality and values vary among animals of the same species. Similarly, there are species-specific variations [...] Read more.

Sperm cryopreservation is one of the sublime biotechnologies for assisted reproduction. In recent decades, there has been an increasing trend in the use of preserved semen. Post-thaw semen quality and values vary among animals of the same species. Similarly, there are species-specific variations in sperm morphology, i.e., sperm head, kinetic properties, plasma membrane integrity, and freezability. Similarly, the viability of sperm varies in the female reproductive tract, i.e., from a few hours (in cattle) to several days (in chicken). Various steps of sperm cryopreservation, i.e., male health examination, semen collection, dilution, semen centrifugation, pre- and post-thaw semen quality evaluation, lack standardized methodology, that result in differences in opinions. Assisted reproductive technologies (ART), including sperm preservation, are not applied to the same extent in commercial poultry species as in mammalian species for management and economic reasons. Sperm preservation requires a reduction in physiological metabolism by extending the viable duration of the gametes. Physiologically and morphologically, spermatozoa are unique in structure and function to deliver paternal DNA and activate oocytes after fertilization. Variations in semen and sperm composition account for better handling of semen, which can aid in improved fertility. This review aims to provide an update on sperm cryopreservation in farm animals. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation)

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