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Cao S, Li S, Lu K, Fu J, Yang C, Qu J, Li J, Zhao X. Peony seed meal supplementation enhances semen quality in aged Qinchuan bulls. Anim Biotechnol 2025; 36:2503752. [PMID: 40380808 DOI: 10.1080/10495398.2025.2503752] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/30/2024] [Accepted: 05/05/2025] [Indexed: 05/19/2025]
Abstract
Healthy Qinchuan bulls aged 8.5-9.5 years were selected and randomly divided into 6 groups based on 0%, 2%, 4%, 6%, 8%, and 10% dietary supplementation of peony seed meal for three months. Overall, linear motility (LM), progressive motility (PM), straight-line velocity (VSL), curvilinear velocity (VCL), and the percentage increase of all sperm grades (A to D) were affected (p<0.05) by the dietary supplementation. LM (29.5 vs. 24.1%), PM (69.4 vs. 60.0%), VLS (38.0 vs. 33.3 μm/s), and VCL (48.3 vs. 44.3 μm/s) were consistently higher in the 6% peony seed meal dietary supplementation groups than in the control group, respectively. A similar pattern was observed in the 8% group. Inversely to sperm grades C and D, grades A and B were higher in all treated groups than the control group, except for the 2% group in sperm grade A. Moreover, the levels of the enzymes Superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR; except for 2% peony seed group), and glutathione peroxidase (GSH-px) were higher than the control group. The level of malondialdehyde (MDA) was lower in all treated groups than in the control group. Dietary supplementation of 2-8% peony seed meal improves the semen quality of aged Qinchuan bulls.
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Affiliation(s)
- Shaojie Cao
- College of Agriculture and Bioengineering, Heze University, Heze, Shandong, China
| | - Shiwei Li
- College of Pharmacy, Heze University, Heze, Shandong, China
| | - Kaixue Lu
- College of Pharmacy, Heze University, Heze, Shandong, China
| | - Juntai Fu
- College of Agriculture and Bioengineering, Heze University, Heze, Shandong, China
| | - Chengwei Yang
- College of Pharmacy, Heze University, Heze, Shandong, China
| | - Jiahui Qu
- College of Agriculture and Bioengineering, Heze University, Heze, Shandong, China
| | - Juanjuan Li
- Weinan Vocational and Technical College, Weinan, Shaanxi, China
| | - Xianlin Zhao
- College of Pharmacy, Heze University, Heze, Shandong, China
- Weinan Vocational and Technical College, Weinan, Shaanxi, China
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Zanganeh Z, Hezavehei M, Halvaei I, Sharafi M. Beneficial Effects of N-Acetyl Cysteine in the Different Equilibration Times on Post-Thawed Rooster Sperm Quality. Reprod Domest Anim 2025; 60:e70035. [PMID: 40056000 DOI: 10.1111/rda.70035] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/01/2024] [Revised: 12/07/2024] [Accepted: 02/27/2025] [Indexed: 05/13/2025]
Abstract
Cryopreservation is the best method for preserving rooster sperm, especially in declining indigenous breeds. Cryopreserved semen is significantly compromised due to equilibration time, cold shock and oxidative stress encountered during the freezing-thawing process. To improve the quality and fertility of thawed semen, it is essential to protect sperm cells from peroxidative damage. This study assessed the effects of N-acetyl cysteine (NAC), an antioxidant supplement, on the functional parameters of thawed rooster sperm after pre-freezing equilibration periods of 2 and 4 h. Samples were collected from 10 male Ross 308 broiler breeders and diluted with Beltsville extenders containing different concentrations of NAC (0, 0.1, 1 and 10 mM/mL) during equilibrium periods of 2 and 4 h before freezing. Our findings showed that NAC-0.1 and NAC-1 groups in 2 h increased significantly total and progressive motility (59.85 ± 3.73, 59.67 ± 3.73, 42.85 ± 2.64 and 42.80 ± 2.64, respectively), viability, and plasma membrane functionality (62.45 ± 3.51, 62.36 ± 3.51, 56.81 ± 3.51 and 56.82 ± 3.56, respectively) compared to the control groups. Furthermore, NAC-0.1-2 h and NAC-1-2 h demonstrated the lowest levels of apoptosis and intracellular reactive oxygen species (ROS), as well as the highest mitochondrial membrane potential in comparison to the control groups. These findings indicate that NAC-0.1 and NAC-1 are effective in maintaining the quality of thawed rooster sperm during a 2-h equilibration period.
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Affiliation(s)
- Zeynab Zanganeh
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
| | - Maryam Hezavehei
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
- Research Center for Reproduction and Fertility, Faculty of Veterinary Medicine, Montreal University, St-Hyacinthe, Quebec, Canada
| | - Iman Halvaei
- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
| | - Mohsen Sharafi
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
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Hussain T, Fayyaz MH, Hameed A, Hassan Andrabi SM, Kausar R, Mubashir Y, Batool I, Shahzad M, Omur AD. Effect of resveratrol on post-thaw motility, kinematics, structural parameters and antioxidant/oxidant status of Kamori buck spermatozoa. Cryobiology 2025; 118:105202. [PMID: 39837404 DOI: 10.1016/j.cryobiol.2025.105202] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2024] [Revised: 01/16/2025] [Accepted: 01/17/2025] [Indexed: 01/23/2025]
Abstract
Resveratrol is a polyphenol compound showing strong antioxidant properties. It is believed that semen cryopreservation causes significant sperm losses which eventually affects sperm quality. Improving antioxidant status of semen may reduce this damage and enhance sperm fertilizing potential. This study investigated the resveratrol treated freeze-thawing sperm motion, kinetics, structural parameters and antioxidant/oxidant status of Kamori buck spermatozoa. Thirty-two ejaculates from 4 fertile Kamori bucks were processed in tris-citric-glyercol-egg yolk (TCG-EY) based with varying concentrations of resveratrol (0, 10, 20, 40 and 50 μM). Over 75 % sperm motility were pooled and frozen in liquid nitrogen. The results unveiled that adding resveratrol at 40 and 50 μM concentration significantly enhanced (P < 0.05) total motility (progressive motility, rapid velocity, and average path velocity, straight line velocity, curvilinear velocity, amplitude of lateral head displacement, beat cross frequency, straightness and linearity in contrast with control and other groups. Supplementing resveratrol at 40 and 50 μM concentration significantly improved (P < 0.05) functional plasma membrane integrity, acrosome integrity, whereas, all resveratrol groups had same significant (P < 0.05) effect on DNA integrity in response to control group. The 40 and 50 μM resveratrol significantly promoted (P < 0.05) superoxide dismutase (SOD), catalase (CAT), peroxidases (POD), and total antioxidant capacity (TAC) levels while significantly reducing (P < 0.05) the total oxidant status (TOS) and malondialdehyde (MDA) contents as compared to control and other groups, respectively. The principal component analysis (PCA) exhibited samples were present in different clusters except two groups which had partially overlapped. Using hierarchical clustering analysis, two clusters were constructed showing the relationship within the treated groups. The results of spearman correlation coefficient showed that yellow color showed highly positive correlation while turquoise color exhibited highly negative association between sperm variables. Overall, the results concluded that resveratrol at 50 μM performed slightly better results than 40 μM in terms of improving sperm quality parameters.
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Affiliation(s)
- Tarique Hussain
- Animal Sciences Division, Nuclear Institute for Agriculture and Biology College, Pakistan Institute of Engineering and Applied Sciences (NIAB-C, PIEAS), Faisalabad, 38000, Pakistan.
| | - Muhammad Hammad Fayyaz
- Animal Sciences Institute, National Agricultural Research Centre, Islamabad, 44000, Pakistan
| | - Amjad Hameed
- Nuclear Institute for Agriculture and Biology College, Pakistan Institute of Engineering and Applied Sciences (NIAB-C, PIEAS), Faisalabad, 38000, Pakistan
| | | | - Rehana Kausar
- Animal Sciences Division, Nuclear Institute for Agriculture and Biology College, Pakistan Institute of Engineering and Applied Sciences (NIAB-C, PIEAS), Faisalabad, 38000, Pakistan
| | - Yasin Mubashir
- Animal Sciences Division, Nuclear Institute for Agriculture and Biology College, Pakistan Institute of Engineering and Applied Sciences (NIAB-C, PIEAS), Faisalabad, 38000, Pakistan
| | - Iqra Batool
- Animal Sciences Division, Nuclear Institute for Agriculture and Biology College, Pakistan Institute of Engineering and Applied Sciences (NIAB-C, PIEAS), Faisalabad, 38000, Pakistan
| | - Muhammad Shahzad
- Animal Sciences Division, Nuclear Institute for Agriculture and Biology College, Pakistan Institute of Engineering and Applied Sciences (NIAB-C, PIEAS), Faisalabad, 38000, Pakistan
| | - Ali Dogan Omur
- Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Ataturk University, Erzurum, Turkiye
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Zong Y, Li Y, Sun Y, Han X, Yuan J, Ma L, Ma H, Chen J. Mitochondrial aspartate aminotransferase (GOT2) protein as a potential cryodamage biomarker in rooster spermatozoa cryopreservation. Poult Sci 2025; 104:104690. [PMID: 39721280 PMCID: PMC11732459 DOI: 10.1016/j.psj.2024.104690] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/22/2024] [Revised: 12/12/2024] [Accepted: 12/17/2024] [Indexed: 12/28/2024] Open
Abstract
Spermatozoa cryopreservation has been widely used for animal genetic conservation. Despite advances in chicken semen cryopreservation, the mechanism of spermatozoa cryodamage remains to be revealed. The cryopreservation process induces motion parameter decreased, structure damaged, proteomic and antioxidant system remodeled in spermatozoa. Mitochondrial glutamate-oxaloacetate transaminase 2 (GOT2) is part of the malate aspartate shuttle, which is ubiquitous in mitochondria and is associated with cellular metabolism regulation. Thus, this study is the first to investigate GOT2 biological role in chicken spermatozoa during freezing process. The results showed that the sperm total motility, straight-line velocity (VSL) and mitochondrial membrane potential (MMP) of the frozen group were significantly lower than these of the fresh group (P < 0.05). The fresh sperm mitochondrial membrane was continuous and mitochondrial matrix was dense and homogeneous. However, after the freezing-thawing, the density of the matrix was reduced, and the mitochondria appeared slightly swollen and membrane damaged. In chicken sperm, the GOT2 protein was localized in the head and the midpiece of spermatozoa where mitochondria are located by immunostaining analysis. This was consistent with the subcellular localization prediction. GOT2 protein was more abundant in the fresh sperm than in the frozen sperm, which indicated that freezing may lead to sperm mitochondrial damage, reduced GOT protein expression, and affected sperm motility and fertility. The protein-protein interaction prediction of GOT2 protein indicated that its ten most confident interactors were predominantly mitochondria-related proteins. The binding ability was higher between GOT2 protein and two mitochondria-targeted antioxidants, SkQ1 and Mito-TEMPO, respectively. In conclusion, GOT2 played an important role in chicken spermatozoa, which was possibly associated with the regulation of mitochondria function and spermatozoa metabolism. Moreover, it may be a potential cryodamage improvement target for spermatozoa. However, the underlying mechanism of GOT2 in spermatozoa cryopreservation needs further exploration.
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Affiliation(s)
- Yunhe Zong
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
| | - Yunlei Li
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
| | - Yanyan Sun
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
| | - Xintong Han
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
| | - Jingwei Yuan
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
| | - Lin Ma
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
| | - Hui Ma
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
| | - Jilan Chen
- State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
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Calisto Bongalhardo D. Sperm fitness assessment in poultry: Brief review of in vitro methods. Anim Reprod Sci 2025; 272:107666. [PMID: 39657511 DOI: 10.1016/j.anireprosci.2024.107666] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2024] [Revised: 11/28/2024] [Accepted: 12/03/2024] [Indexed: 12/12/2024]
Abstract
In the poultry industry, every chick or poult produced has high economic value; therefore, selection of male breeders capable of transmitting desirable traits to their offspring has an important role in fertility and hatching success. Following either natural mating or artificial insemination, sperm must go through a sequence of steps to fertilize the egg: 1. Go from the site of deposition (vagina) to the Sperm Storage Tubules (SST); 2. Enter and stay inside the SST; 3. Reacquire motility; and 4. Bind to and penetrate the egg. To perform these tasks successfully, sperm must be alive, motile, and have structural integrity (membrane, glycocalyx, acrosome, and DNA). To evaluate sperm fitness, several in vitro tests can be performed; however, it is often necessary to combine several assays to have a more reliable fertility estimation, as each procedure evaluates specific and distinct sperm features. In this article, we briefly review in vitro tests that can be used to evaluate poultry sperm, associating them with each step for fertilization. The sperm-egg interaction assay using the inner perivitelline layer of chicken eggs is highlighted, since it is a single test that evaluates multiple sperm characteristics and is highly correlated with fertility.
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Hakimi F, Karimi Torshizi MA, Hezavehei M, Sharafi M. Protective Effect of N-Acetylcysteine on Rooster Semen Cryopreservation. Biopreserv Biobank 2024; 22:609-615. [PMID: 38634668 PMCID: PMC11656127 DOI: 10.1089/bio.2023.0103] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 04/19/2024] Open
Abstract
Cryopreservation of avian semen is a useful reproductive technique in the poultry industry. However, during cooling, elevated reactive oxygen species (ROS) levels have destructive effects on both quality and function of thawed sperm. The aim of the current study is to investigate the antioxidant effects of N-acetylcysteine (NAC) during rooster semen cryopreservation. Semen samples were collected from ten Ross 308 broiler breeder roosters (32 weeks) and mixed. The mixed samples were divided into five equal parts and cryopreserved in Lake Buffer extender that contained different concentrations (0, 0.01, 0.1, 1, and 10 mM) of NAC. The optimum concentration of NAC was determined based on quality parameters of mobility, viability, membrane integrity, acrosome integrity, lipid peroxidation, and mitochondrial membrane potential after the freeze-thaw process. There was a higher percentage (p < 0.05) of total motility (TM) (60.9 ± 2.4%) and progressive motility (PM) (35.6 ± 1.9%) observed with the NAC-0.1 group compared to the other groups. Significantly higher percentages of viability (74.4 ± 2.3% and 71 ± 2.3%), membrane integrity (76.4 ± 1.5% and 74.7 ± 1.5%) and mitochondrial membrane potential (67.1 ± 1.6% and 66.3 ± 1.6%) were observed in the NAC-0.1 and NAC-1 groups compared to the other frozen groups (p < 0.05). The lowest percentage of lipid peroxidation and nonviable sperm was found in the NAC-0.1 and NAC-1 groups compared to the other groups (p < 0.05). The average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), and acrosome integrity, were not affected by different concentrations of NAC in the thawed sperm (p > 0.05). Both NAC-0.1 and NAC-1 appear to be beneficial for maintaining the quality of rooster sperm after thawing.
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Affiliation(s)
- Farhad Hakimi
- Department of Animal Sciences, College of Agriculture, Tarbiat Modares University, Tehran, Iran
| | | | - Maryam Hezavehei
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
- Research Center for Reproduction and Fertility, Faculty of Veterinary medicine, Montreal University, St-Hyacinthe, Canada
| | - Mohsen Sharafi
- Department of Animal Sciences, College of Agriculture, Tarbiat Modares University, Tehran, Iran
- Semex Alliance, Guelph, Canada
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Díaz Ruiz E, Delgado Bermejo JV, León Jurado JM, Navas González FJ, Arando Arbulu A, Fernández-Bolaños Guzmán J, Bermúdez Oria A, González Ariza A. Effect of Supplementation of a Cryopreservation Extender with Pectoliv30 on Post-Thawing Semen Quality Parameters in Rooster Species. Antioxidants (Basel) 2024; 13:1018. [PMID: 39199262 PMCID: PMC11351633 DOI: 10.3390/antiox13081018] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2024] [Revised: 08/19/2024] [Accepted: 08/19/2024] [Indexed: 09/01/2024] Open
Abstract
Sperm cryopreservation is a fundamental tool for the conservation of avian genetic resources; however, avian spermatozoa are susceptible to this process. To cope with the high production of reactive oxygen species (ROS), the addition of exogenous antioxidants is beneficial. Pectoliv30 is a substance derived from alperujo, and in this study, its effect was analyzed on seminal quality after its addition to the cryopreservation extender of roosters at different concentrations. For this purpose, 16 Utrerana breed roosters were used, and seminal collection was performed in six replicates, creating a pool for each working day with ejaculates of quality. After cryopreservation, one sample per treatment and replicate was thawed, and several seminal quality parameters were evaluated. Statistical analysis revealed numerous correlations between these variables, both positive and negative according to the correlation matrix obtained. Furthermore, the chi-squared automatic interaction detection (CHAID) decision tree (DT) reported significant differences in the hypo-osmotic swelling test (HOST) variable between groups. Moreover, results for this parameter were more desirable at high concentrations of Pectoliv30. The application of this substance extracted from the by-product alperujo as an antioxidant allows the improvement of the post-thawing seminal quality in roosters and facilitates optimization of the cryopreservation process as a way to improve the conservation programs of different endangered poultry breeds.
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Affiliation(s)
- Esther Díaz Ruiz
- Department of Genetics, Faculty of Veterinary Sciences, University of Córdoba, 14071 Cordoba, Spain; (E.D.R.); (J.V.D.B.); (A.A.A.)
- Institute of Agricultural Research and Training (IFAPA), Alameda del Obispo, 14005 Cordoba, Spain
| | - Juan Vicente Delgado Bermejo
- Department of Genetics, Faculty of Veterinary Sciences, University of Córdoba, 14071 Cordoba, Spain; (E.D.R.); (J.V.D.B.); (A.A.A.)
| | | | - Francisco Javier Navas González
- Department of Genetics, Faculty of Veterinary Sciences, University of Córdoba, 14071 Cordoba, Spain; (E.D.R.); (J.V.D.B.); (A.A.A.)
| | - Ander Arando Arbulu
- Department of Genetics, Faculty of Veterinary Sciences, University of Córdoba, 14071 Cordoba, Spain; (E.D.R.); (J.V.D.B.); (A.A.A.)
| | - Juan Fernández-Bolaños Guzmán
- Instituto de la Grasa, Consejo Superior de Investigaciones Científicas (CSIC), 41013 Sevilla, Spain; (J.F.-B.G.); (A.B.O.)
| | - Alejandra Bermúdez Oria
- Instituto de la Grasa, Consejo Superior de Investigaciones Científicas (CSIC), 41013 Sevilla, Spain; (J.F.-B.G.); (A.B.O.)
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Kargari M, Sharafi M, Torshizi MAK, Hezavehei M, Zanganeh Z. Effects of hydroxytyrosol on post-thaw quality of rooster sperm. Reprod Domest Anim 2024; 59:e14588. [PMID: 38822558 DOI: 10.1111/rda.14588] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/11/2023] [Revised: 04/02/2024] [Accepted: 04/29/2024] [Indexed: 06/03/2024]
Abstract
Semen cryopreservation is one of the most important reproduction techniques in the livestock and poultry industry. Cryopreservation induces cold stress, generating reactive oxygen species (ROS) and oxidative stress causing structural and biochemical damages in sperm. In this study, we evaluated the effects of the hydroxytyrosol (HT), as an antioxidant, at the concentrations of 0, 25, 50, and 100 μg/mL on post-thaw semen quality metrics in rooster. Semen samples were collected twice a week from 10 roosters (29 weeks), processed and frozen according to experimental groups. Different quality parameters, including total motility, progressive motility, viability, morphology, membrane integrity, and malondialdehyde were measured after thawing. Results showed that 25 and 50 μg/mL of HT produced the highest percentage of total motility (51.01 ± 2.19 and 50.15 ± 2.19, respectively) and progressive motility (35.74 ± 1.34 and 35.15 ± 1.34, respectively), membrane integrity (48.00 ± 2.18 and 46.75 ± 2.18, respectively) as well as viability (53.00 ± 2.17 and 52.50 ± 2.17, respectively) compared with the other groups (p < .05). The group with 25 μg/mL of HT showed the lowest significant (p < .05) MDA concentration (1.81 ± 0.25). Our results showed that the effect of HT was not dose-dependent and optimum concentration of HT could improve functional parameters of rooster sperm after freezing-thawing. These findings suggest that HT may have protective effects on the rooster sperm during the freezing-thawing process.
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Affiliation(s)
- Mohammad Kargari
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
| | - Mohsen Sharafi
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
| | | | - Maryam Hezavehei
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Zeynab Zanganeh
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
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Díaz Ruiz E, González Ariza A, León Jurado JM, Arando Arbulu A, Fernández-Bolaños Guzmán J, Bermúdez Oria A, Delgado Bermejo JV, Navas González FJ. Evaluation of the effect of the addition of an olive oil-derived antioxidant (Pectoliv-80A) in the extender for cryopreservation of rooster sperm through the use of a discriminant statistical tool. Poult Sci 2024; 103:103630. [PMID: 38513548 PMCID: PMC10973192 DOI: 10.1016/j.psj.2024.103630] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/05/2024] [Revised: 03/02/2024] [Accepted: 03/04/2024] [Indexed: 03/23/2024] Open
Abstract
During the poultry sperm cryopreservation process, an excess of reactive oxygen species is generated resulting in oxidative stress which harms the quality of avian spermatozoa. To counteract this effect, the addition of exogenous antioxidants, such as Pectoliv-80A (a by-product of olive oil), to the cryopreservation diluent is interesting. For this purpose, 16 roosters belonging to the Utrerana avian breed were used. Six semen pools (from the 6 different replicates) were divided into 4 aliquots corresponding to different concentrations of Pectoliv-80A that were tested (0, 300, 400, and 500 μg/mL), and the cryopreservation process was carried out. To evaluate post-thawing semen quality, different parameters such as motility, membrane functionality, reactive oxygen species production, lipid peroxidation, and acrosome integrity were studied. A discriminant canonical analysis was used to determine both the differences between the Pectoliv-80A concentration groups and the discriminant power of the aforementioned parameter used for semen evaluation. Total motility and membrane functionality were reported to be the most discriminant variables for differentiating the different antioxidant enrichment groups and concluded that concentrations of 300 μg/mL showed the most desirable quality of post-thawing semen. The present study could lead to the optimization of both cryopreservation and quality evaluation techniques of the sperm of rooster species, that support the conservation program of endangered local breeds.
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Affiliation(s)
- Esther Díaz Ruiz
- Department of Genetics, Faculty of Veterinary Sciences, University of Córdoba, Córdoba, 14071, Spain
| | | | | | - Ander Arando Arbulu
- Department of Genetics, Faculty of Veterinary Sciences, University of Córdoba, Córdoba, 14071, Spain
| | | | - Alejandra Bermúdez Oria
- Instituto de la Grasa, Consejo Superior de Investigaciones Científicas, (CSIC), Sevilla, 41013, Spain
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Baharsaadi M, Hezavehei M, Shahverdi A, Halvaei I. Evaluation of the effects of hydroxytyrosol on human sperm parameters during cryopreservation. Cryobiology 2024; 114:104840. [PMID: 38104853 DOI: 10.1016/j.cryobiol.2023.104840] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/19/2023] [Revised: 10/31/2023] [Accepted: 12/11/2023] [Indexed: 12/19/2023]
Abstract
Human sperm cryopreservation is a routine procedure in assisted reproductive technology, but it has detrimental effects on different sperm parameters due to oxidative stress. Our objective was to assess the impacts of hydroxytyrosol (HT), as an antioxidant, on human sperm parameters following cryopreservation. In the first phase, 20 normal human semen samples were cryopreserved using the rapid freezing method with different concentrations of HT including 0, 50, 100, 150, and 200 μg/mL. In the second phase, 20 normal semen samples were collected and cryopreserved with 50 and 100 μg/mL HT. The beneficial effects of HT were determined by evaluation of motility (computer-assisted sperm analysis; CASA), viability (Eosin-nigrosine stain), DNA integrity (sperm chromatic dispersion test, SCD), reactive oxygen species (DCF and DHE staining by flowcytometry) lipid peroxidation (malondialdehyde, MDA test) and mitochondrial membrane potential (JC1 staining by flowcytometry) of sperm after cryopreservation. After thawing, sperm motility had an increasing trend in 50 and 100 μg/mL HT groups in comparison with other groups, althought the difference was not significant. However, sperm viability was significantly increased at 50 and 100 μg/mL HT. Our data also showed that sperm DNA fragmentation was significantly decreased after thawing at 100 μg/mL in comparison with 0 and 50 μg/mL HT. However, the level of intracellular reactive oxygen species, lipid peroxidation and mitochondrial membrane potential were not significantly different between groups. Our results showed that HT may have protective effects on the viability and DNA integrity of human sperm during the freezing-thawing process.
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Affiliation(s)
- Mojtaba Baharsaadi
- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
| | - Maryam Hezavehei
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
| | - Abdolhossein Shahverdi
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
| | - Iman Halvaei
- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
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11
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Zhu Z, Zhao H, Cui H, Adetunji AO, Min L. Resveratrol Improves the Frozen-Thawed Ram Sperm Quality. Animals (Basel) 2023; 13:3887. [PMID: 38136923 PMCID: PMC10740518 DOI: 10.3390/ani13243887] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/19/2023] [Revised: 12/14/2023] [Accepted: 12/15/2023] [Indexed: 12/24/2023] Open
Abstract
Cryopreservation generates a substantial quantity of ROS in semen, leading to a decline in sperm quality and fertilization capacity. The objective of this study was to investigate the effects of resveratrol and its optimal concentration on ram sperm quality after cryopreservation. Ram semen was diluted with a freezing medium containing different concentrations of resveratrol (0, 25, 50, 75, and 100 μM). After thawing, various sperm parameters such as total motility, progressive motility, acrosome integrity, plasma membrane integrity, mitochondrial membrane potential, glutathione (GSH) content, glutathione synthase (GPx) activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, lipid peroxidation (LPO) content, malondialdehyde (MDA) content, ROS level, SIRT1 level, DNA oxidative damage, and AMPK phosphorylation level were assessed. In addition, post-thaw sperm apoptosis was evaluated. Comparatively, the addition of resveratrol up to 75 μM significantly improved the sperm motility and sperm parameters of cryopreserved ram sperm. Specifically, 50 μM resveratrol demonstrated a notable enhancement in acrosome and plasma membrane integrity, antioxidant capacity, mitochondrial membrane potential, adenosine triphosphate (ATP) content, SIRT1 level, and AMPK phosphorylation levels compared to the control group (p < 0.05). It also significantly (p < 0.05) reduced the oxidative damage to sperm DNA. However, detrimental effects of resveratrol were observed at a concentration of 100 μM resveratrol. In conclusion, the addition of 50 μM resveratrol to the cryopreservation solution is optimal for enhancing the quality of cryopreserved ram sperm.
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Affiliation(s)
- Zhendong Zhu
- College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China; (Z.Z.); (H.Z.); (H.C.)
| | - Haolong Zhao
- College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China; (Z.Z.); (H.Z.); (H.C.)
| | - Haixiang Cui
- College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China; (Z.Z.); (H.Z.); (H.C.)
| | - Adedeji O. Adetunji
- Department of Agriculture, University of Arkansas at Pine Bluff, Pine Bluff, AR 71601, USA;
| | - Lingjiang Min
- College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China; (Z.Z.); (H.Z.); (H.C.)
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12
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Farrokhi Z, Sharafi M, Hezavehei M, Torabi A, Shahverdi M, Rahimi S. The Effects of Glycerophospholipid Nanomicelles on the Cryotolerance of Frozen-Thawed Rooster Sperm. Biopreserv Biobank 2023; 21:593-598. [PMID: 36637861 DOI: 10.1089/bio.2022.0135] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/14/2023] Open
Abstract
Semen banking is an efficient method of artificial insemination for commercial breeders. However, the cryopreservation process induces severe damages to plasma membranes, which leads to reduced fertility potential of thawed sperm. The replacement of membrane lipids with oxidized membrane lipids repairs the cell membrane and improves its stability. The aim of this study was to investigate the effects of glycerophospholipid (GPL) nanomicelles on the cryosurvival of thawed rooster semen. Semen samples were collected from six 29-week Ross broiler breeder roosters, then mixed and divided into five equal parts. The samples were diluted with the Beltsville extender containing different concentrations of GPL according to the following groups: 0 (GPL-0), 0.1% (GPL-0.1), 0.5% (GPL-0.5), 1% (GPL-1), and 1.5% (GPL-1.5), then diluted semen was gradually cooled to 4°C during 3 hours and stored in liquid nitrogen. The optimum concentration of GPL was determined based on the quality parameters of thawed sperm. Our results showed sperm exposed to GPL-1 had significantly increased motion parameters and mitochondrial activity. The percentages of viability and membrane integrity were significantly higher in the GPL-1, and GPL-1.5 groups compared with the other groups (p < 0.05). Moreover, the lowest rate of apoptosis and lipid peroxidation were observed in the GPL-1 and GPL-1.5 groups in comparison with the frozen control group. Our findings indicated that membrane lipid replacement with GPL nanomicelles (1% and 1.5%) could substitute for damaged lipids in membranes and protect sperm cells against cryoinjury.
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Affiliation(s)
- Zahra Farrokhi
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
- Department of Embryology Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Mohsen Sharafi
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
- Department of Embryology Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Maryam Hezavehei
- Department of Embryology Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Ali Torabi
- Research Center for Reproduction and Fertility, Faculty of Veterinary medicine, Montreal University, St-Hyacinthe, Canada
| | - Maryam Shahverdi
- Department of Embryology Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Shaban Rahimi
- Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
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13
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Nguyen HT, Do SQ, Athurupana R, Wakai T, Funahashi H. Rapid thawing of frozen bull spermatozoa by transient exposure to 70 °C improves the viability, motility and mitochondrial health. Anim Reprod 2023; 20:e20220127. [PMID: 38026001 PMCID: PMC10681132 DOI: 10.1590/1984-3143-ar2022-0127] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2022] [Accepted: 07/13/2023] [Indexed: 12/01/2023] Open
Abstract
Up to now, the definitive conclusion of the positive effects of rapid transient thawing at higher temperatures for shorter durations has not been obtained yet and is still under discussion due to some contradictory findings and limited assessment of post-thawed parameters. The purpose of the current study was to evaluate the effectiveness of rapid thawing in water at 70 °C by using various post-thawed parameters of frozen bull spermatozoa. Experiment 1, monitoring the change of temperature inside frozen bull straw thawed in water at different temperatures. Experiment 2, evaluation of various post-thawed characteristics of frozen bull spermatozoa thawed in water at different temperatures by using a computer-assisted sperm analysis, flow cytometry and immunocytochemistry. The time it took for the temperature inside the straw to warm up to 15 °C was nearly twice as faster when the straw was thawed in 70 °C water compared with 39 °C. Although there were differences among bulls, viability, motility, and mitochondrial membrane potential of spermatozoa thawed at 70 °C for 8 seconds and stabilized at 39 °C for 52 seconds were significantly higher than those of controls (thawed at 39 °C for 60 seconds) at 0 and 3 h after thawing. Just after thawing, however, there were no differences in acrosome integrity and distribution of phospholipase C zeta1, whereas mitochondrial reactive oxygen species production was significantly lower in spermatozoa thawed at 70 °C. From these results, we conclude that rapid thawing at 70 °C and then stabilization at 39 °C significantly improves viability, motility and mitochondrial health of bull spermatozoa rather than conventional thawing at 39 °C. The beneficial effect of rapid transient thawing could be due to shorter exposure to temperatures outside the physiological range, consequently maintaining mitochondrial health.
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Affiliation(s)
- Hai Thanh Nguyen
- Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan
- Department of Animal Production, Faculty of Animal Science and Veterinary Medicine, Nong Lam University, Ho Chi Minh City, Vietnam
| | - Son Quang Do
- Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan
| | - Rukmali Athurupana
- Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan
| | - Takuya Wakai
- Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan
| | - Hiroaki Funahashi
- Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan
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14
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Kaeoket K, Chanapiwat P. The Beneficial Effect of Resveratrol on the Quality of Frozen-Thawed Boar Sperm. Animals (Basel) 2023; 13:2829. [PMID: 37760229 PMCID: PMC10526068 DOI: 10.3390/ani13182829] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/31/2023] [Revised: 08/22/2023] [Accepted: 08/28/2023] [Indexed: 09/29/2023] Open
Abstract
This study aimed to determine the effect of resveratrol and its optimal concentration on the quality of frozen-thawed (FT) boar sperm. Semen ejaculates were obtained from 13 Duroc boars aged between 1.5 and 3 years. The sperm sample was separated into 7 groups based on the concentrations of resveratrol in the freezing extender, which were 0 (control), 25, 50, 75, 100, 125, and 250 µM, respectively. The sperm was frozen using liquid nitrogen vapor and thawed at 50 °C for 12 s. After thawing, total motility, progressive motility, viability, intact acrosomes, mitochondrial membrane potential and level of MDA were assessed. The supplementation of 50-100 µM resveratrol improved the sperm motility and viability of FT sperm in comparison to the control group (p < 0.05). Furthermore, the 50 µM resveratrol group was significantly more protective than the control group in terms of intact acrosome, mitochondrial membrane potential, and level of MDA (p < 0.05). Nonetheless, the detrimental effect of resveratrol was found at a concentration of 250 µM. In conclusion, the addition of 50-100 µM resveratrol to a freezing extender is the optimal concentration for enhancing the quality of cryopreserved boar sperm.
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Affiliation(s)
| | - Panida Chanapiwat
- Semen Laboratory, Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University, Nakhon-Pathom 73170, Thailand;
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15
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Ratchamak R, Authaida S, Koedkanmark T, Boonkum W, Semaming Y, Chankitisakul V. Supplementation of Freezing Medium with Ginseng Improves Rooster Sperm Quality and Fertility Relative to Free Radicals and Antioxidant Enzymes. Animals (Basel) 2023; 13:2660. [PMID: 37627452 PMCID: PMC10451814 DOI: 10.3390/ani13162660] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/27/2023] [Revised: 08/17/2023] [Accepted: 08/17/2023] [Indexed: 08/27/2023] Open
Abstract
To the best of our knowledge, this study is the first to determine the effect of ginseng as an antioxidant supplement in freezing extenders on the quality of cryopreserved rooster semen. Semen samples were collected from 40 Thai native roosters (Pradu Hang Dum) using the dorso-abdominal massage method and then pooled and divided into five groups according to the concentrations of ginseng supplementation (0, 0.25, 0.50, 0.75, and 1 mg/mL) in a freezing extender. The semen suspensions were loaded into a medium straw and cryopreserved using the liquid nitrogen vapor method. The post-thaw semen was evaluated for sperm quality (sperm motility and membrane integrity), seminal plasma characteristics (lipid peroxidation, superoxide dismutase [SOD], catalase [CAT], and glutathione peroxidase [GPx]), and fertility. The results showed that ginseng extract supplementation at 0.25 mg/mL yielded the highest total motility, progressive motility, and membrane integrity (59.47%, 30.82%, and 48.30%, respectively; p < 0.05) in cryopreserved rooster semen. Higher malondialdehyde concentrations were observed in the control group than in the other groups (p < 0.05). SOD, CAT, and GPx increased compared with those in the control group (p < 0.05). The results showed that the fertility rate with 0.25 mg/mL of ginseng was higher than that of the control group (62.80% vs. 46.28%: p < 0.05). In conclusion, supplementation with 0.25 mg/mL of ginseng is recommended as an alternative component to the freezing extender to improve rooster semen cryopreservation.
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Affiliation(s)
- Ruthaiporn Ratchamak
- Department of Animal Science, Faculty of Agricultural, Khon Kaen University, Khon Kaen 40002, Thailand; (R.R.); (S.A.); (T.K.); (W.B.)
- The Research and Development Network Center of Animal Breeding and Omics, Khon Kaen University, Khon Kaen 40002, Thailand
| | - Supakorn Authaida
- Department of Animal Science, Faculty of Agricultural, Khon Kaen University, Khon Kaen 40002, Thailand; (R.R.); (S.A.); (T.K.); (W.B.)
| | - Thirawat Koedkanmark
- Department of Animal Science, Faculty of Agricultural, Khon Kaen University, Khon Kaen 40002, Thailand; (R.R.); (S.A.); (T.K.); (W.B.)
| | - Wuttigrai Boonkum
- Department of Animal Science, Faculty of Agricultural, Khon Kaen University, Khon Kaen 40002, Thailand; (R.R.); (S.A.); (T.K.); (W.B.)
- The Research and Development Network Center of Animal Breeding and Omics, Khon Kaen University, Khon Kaen 40002, Thailand
| | - Yoswaris Semaming
- Program in Veterinary Technology, Faculty of Technology, Udon Thani Rajabhat University, Udon Thani 41000, Thailand;
| | - Vibuntita Chankitisakul
- Department of Animal Science, Faculty of Agricultural, Khon Kaen University, Khon Kaen 40002, Thailand; (R.R.); (S.A.); (T.K.); (W.B.)
- The Research and Development Network Center of Animal Breeding and Omics, Khon Kaen University, Khon Kaen 40002, Thailand
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16
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Jurado-Campos A, Soria-Meneses PJ, Arenas-Moreira M, Alonso-Moreno C, Rodríguez-Robledo V, Soler AJ, Garde JJ, Del Rocío Fernández-Santos M. Minimizing sperm oxidative stress using nanotechnology for breeding programs in rams. J Anim Sci Biotechnol 2023; 14:106. [PMID: 37559077 PMCID: PMC10413538 DOI: 10.1186/s40104-023-00907-3] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/09/2023] [Accepted: 06/12/2023] [Indexed: 08/11/2023] Open
Abstract
BACKGROUND Artificial insemination (AI) is a routine breeding technology in animal reproduction. Nevertheless, the temperature-sensitive nature and short fertile lifespan of ram sperm samples hamper its use in AI. In this sense, nanotechnology is an interesting tool to improve sperm protection due to the development of nanomaterials for AI, which could be used as delivery vehicles. In this work, we explored the feasibility of vitamin E nanoemulsion (NE) for improving sperm quality during transport. RESULTS With the aim of evaluating this proposal, ejaculates of 7 mature rams of Manchega breed were collected by artificial vagina and extended to 60 × 106 spz/mL in Andromed®. Samples containing control and NE (12 mmol/L) with and without exogenous oxidative stress (100 µmol/L Fe2+/ascorbate) were stored at 22 and 15 ºC and motility (CASA), viability (YO-PRO/PI), acrosomal integrity (PNA-FITC/PI), mitochondrial membrane potential (Mitotracker Deep Red 633), lipoperoxidation (C11 BODIPY 581/591), intracellular reactive oxygen species (ROS) production and DNA status (SCSA®) monitored during 96 h. Our results show that NE could be used to maintain ram spermatozoa during transport at 15 and 22 ºC for up to 96 h, with no appreciable loss of kinematic and physiological characteristics of freshly collected samples. CONCLUSIONS The storage of ram spermatozoa in liquid form for 2-5 d with vitamin E nanoemulsions may lead more flexibility to breeders in AI programs. In view of the potential and high versatility of these nanodevices, further studies are being carried out to assess the proposed sperm preservation medium on fertility after artificial insemination.
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Affiliation(s)
| | | | - María Arenas-Moreira
- Departamento de Química Inorgánica, Orgánica Y Bioquímica-Centro de Innovación en Química Avanzada (ORFEO-CINQA), Facultad de Farmacia, Universidad de Castilla-La Mancha, 02008, Albacete, Spain
- Centro Regional de Investigación Biomédicas, Unidad nanoDrug, Universidad de Castilla-La Mancha, 02008, Albacete, Spain
| | - Carlos Alonso-Moreno
- Departamento de Química Inorgánica, Orgánica Y Bioquímica-Centro de Innovación en Química Avanzada (ORFEO-CINQA), Facultad de Farmacia, Universidad de Castilla-La Mancha, 02008, Albacete, Spain
- Centro Regional de Investigación Biomédicas, Unidad nanoDrug, Universidad de Castilla-La Mancha, 02008, Albacete, Spain
| | - Virginia Rodríguez-Robledo
- SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, S/N, 02071, Albacete, Spain
- Facultad de Farmacia, Universidad de Castilla La Mancha, 02071, Albacete, Spain
| | - Ana Josefa Soler
- SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, S/N, 02071, Albacete, Spain
| | - José Julián Garde
- SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, S/N, 02071, Albacete, Spain
| | - María Del Rocío Fernández-Santos
- SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, S/N, 02071, Albacete, Spain.
- Facultad de Farmacia, Universidad de Castilla La Mancha, 02071, Albacete, Spain.
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17
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Sharafi M, Blondin P, Vincent P, Anzar M, Benson JD. Hydroxytyrosol and resveratrol improves kinetic and flow cytometric parameters of cryopreserved bull semen with low cryotolerance. Anim Reprod Sci 2022; 245:107065. [PMID: 36115090 DOI: 10.1016/j.anireprosci.2022.107065] [Citation(s) in RCA: 6] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2021] [Revised: 08/29/2022] [Accepted: 08/31/2022] [Indexed: 11/15/2022]
Abstract
There is considerable interest in breeding programs to "rescue" semen with poor post-thaw fertility from bulls known as "bad freezers". We hypothesized that there may be an interaction between the post-thaw recovery of sperm and the efficacy of antioxidant addition to extenders. The current study assesses the effects of antioxidant additives hydroxytyrosol (HT) and resveratrol (RSV) on the post-thaw semen parameters in two groups of bulls classified as either high or low cryotolerant (i.e., "good" and "bad" freezers). Semen samples were collected from 40 bulls and processed in the extenders containing different concentrations of HT (experiment 1; 0, 25 and 50 µM) and RSV (experiment 2; 0.0, 0.01, 0.1 and 1 mM). In experiment 1, bulls in the low cryotolerance group had a significant improvement in post-thaw recovery at 25 µM and 50 µM (P < 0.05). These improvements were observed in motility and several cellular parameters. However, post-thaw semen quality in the high cryotolerance group was not significantly affected by the HT addition. In experiment 2, although RSV did not have any positive impact in high cryotolerance group, post-thaw recovery in the low cryotolerance bulls was significantly improved in 0.1 mM RSV. Oxidative stress markers in either high or low cryotolerance groups were not significantly changed by RSV addition (P > 0.05). It can be concluded that addition of optimized concentrations of HT and RSV to the extender could be a strategy for improving the post-thaw semen, especially in bulls with high genetic merit but low initial cryotolerance.
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Affiliation(s)
- Mohsen Sharafi
- Department of Biology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada
| | | | | | - Muhammad Anzar
- Agriculture and AgriFood Canada, Saskatoon Research and Development Center, and Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada
| | - James D Benson
- Department of Biology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
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18
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Mehdipour M, Daghigh-Kia H, Najafi A, Mehdipour Z, Mohammadi H. Protective effect of rosiglitazone on microscopic and oxidative stress parameters of ram sperm after freeze-thawing. Sci Rep 2022; 12:13981. [PMID: 35978030 PMCID: PMC9385643 DOI: 10.1038/s41598-022-18298-2] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/20/2022] [Accepted: 08/09/2022] [Indexed: 11/29/2022] Open
Abstract
The purpose of this study was to investigate the effects of rosiglitazone on ram semen after cryopreservation on the quality of thawed sperm. Sperm motility, membrane functionality, viability, total abnormality, acrosome membrane integrity, mitochondrial activity, reactive oxygen species production, ATP content and apoptotic features were assessed after thawing. Rosiglitazone at concentration of 60 µM resulted in the highest (P < 0.05) total motility, progressive motility and straight-line velocity. The percentages of average path velocity and curvilinear velocity were greater in the 60 µM group. Different concentrations of rosiglitazone did not have significant effects on amplitude of the lateral head displacement, linearity and straightness. The highest amounts of membrane functionality and mitochondrial activity after freeze-thawing were observed in groups containing 60 µM. By increasing the rosiglitazone level to 80 µM, no positive effect was observed in most of the evaluated parameters. The lowest ROS concentration was recorded in 60 µM rosiglitazone group (P < 0.05). The group containing 60 µM rosiglitazone also produced the lowest significant percentage of apoptosis-like changes and dead sperm. A greater (P < 0.05) percentage of acrosome integrity in frozen-thawed spermatozoa was observed in the 60 µM rosiglitazone group. There was no significant difference between 40 and 60 µM rosiglitazone in intact acrosome of ram thawed semen. The result showed that supplementation in ram semen extender with rosiglitazone had a positive role in the regulation of ram sperm motility and had strong protective effect on the sperm membrane and acrosome integrity.
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Affiliation(s)
- Mahdieh Mehdipour
- Department of Animal Science, College of Agriculture, University of Tabriz, Tabriz, Iran
| | - Hossein Daghigh-Kia
- Department of Animal Science, College of Agriculture, University of Tabriz, Tabriz, Iran.
| | - Abouzar Najafi
- Department of Animal and Poultry Science, College of Aburaihan, University of Tehran, Tehran, Iran
| | - Zohreh Mehdipour
- Prestage Department of Poultry Science, North Carolina State University, Raleigh, NC, 27606, USA
| | - Hossein Mohammadi
- Department of Animal Science, Faculty of Agriculture and Natural Resources, Arak University, Arak, Iran
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19
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Jimoh OA, Nwachukwu CU. Efficacy of soursop juice extender on rooster semen quality, oxidative activity and spermatozoa kinematics. Transl Anim Sci 2022; 6:txac046. [PMID: 35599843 PMCID: PMC9119429 DOI: 10.1093/tas/txac046] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/19/2022] [Accepted: 04/08/2022] [Indexed: 12/03/2022] Open
Abstract
African medicinal plant like soursop (Annona muricate L.) within annonaceae are known for their biological, therapeutic, and pharmacological properties with little or no toxicity. The use of such plant requires good knowledge of the toxicity dosage, purity, suitable extraction solvent and adverse effects. The leaves, seeds, fruits, barks, and roots of African medicinal plants have been used for various nutraceuticals and functional effects according to African folk medicine. The aim of this study is to evaluate the semen quality, oxidative activity and spermatozoa kinematics of rooster semen in soursop juice extender. About 30 roosters were used for the in vitro analysis. Semen was collected twice a week for 2 weeks through dorsal-abdominal massage technique. The evaluation was done hourly until semen quality declined at the 5th-hour. The pooled semen was allotted to seven treatments of semen extenders as undiluted semen, dextrose saline, 10% soursop juice extender, 20% soursop juice extender, 30% soursop juice extender, 40% soursop juice extender, and 50% soursop juice extender for the study. The percentage motility, progressive motility, nonprogressive motility, curvilinear velocity, average path velocity, straight line velocity, linearity, straightness, amplitude of lateral head, beat cross frequency and wobble were analyzed using computer aided sperm analysis. Oxidative status (antioxidant activity and lipid peroxidation) was determined by assay. Result of rooster semen at room temperature and after 1-hour dilution showed that percentage motility, nonprogressive motility, and average path velocity were significantly (P < 0.05) reduced by different soursop juice extenders compared to undiluted semen. After 2-hour dilution of rooster semen, nonprogressive motility, average path velocity, curvilinear velocity, straight line velocity, wobble, liveability and amplitude of lateral head parameters were significantly (P < 0.05) increased by different soursop juice extenders compared to undiluted semen. Antioxidant activity and lipid peroxidation in both room temperature and after 5-hour dilution were affected by different soursop juice extenders in rooster semen. In conclusion, supplementation of soursop juices as an extender to rooster undiluted semen played an improvement role on spermatozoa fertility and oxidative status during processing or preserving ejaculates for insemination.
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Affiliation(s)
- Olatunji Abubakar Jimoh
- Agricultural Technology Department, Federal Polytechnic Ado Ekiti, P. M. B. 5351, Ado Ekiti, Ekiti State, Nigeria
| | - Chinwe Uchechi Nwachukwu
- Department of Agricultural Science Education, School of Vocational and Technical Education, Alvan Ikoku Federal College of Education, Owerri, Imo State, Nigeria
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