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14 pages, 2189 KiB

Open AccessArticle

Effects of Extenders Supplementation with Gum Arabic and Antioxidants on Ram Spermatozoa Quality after Cryopreservation

by Mohamed Shehab-El-Deen, Mohamed Ali and Mohammed Al-Sharari

Animals 2023, 13(1), 111; https://doi.org/10.3390/ani13010111 - 28 Dec 2022

Cited by 6 | Viewed by 2470

Abstract

Semen cryopreservation is very important in animal agriculture to maximize the number of daughters of genetically superior males and to distribute the cryopreserved semen of good males all over the world. However, the freezing process generates some damage to sperm that reduce their [...] Read more.

Semen cryopreservation is very important in animal agriculture to maximize the number of daughters of genetically superior males and to distribute the cryopreserved semen of good males all over the world. However, the freezing process generates some damage to sperm that reduce their fertilizing ability after thawing. Moreover, egg yolk, which is the most common animal-origin cryoprotectant used in semen dilution, is considered a source of biosecurity risk. In the current study, we aimed to compare the replacement of egg yolk in the extender by gum arabic (5%) along with supplementation with antioxidant cysteine or ascorbic acid on semen quality and freezability in Noemi rams in vitro. Semen from six rams were collected with an artificial vagina two times per week. Semen evaluation parameters such as color, volume, pH, general motility, percentage motility, concentration and cell viability ratio were assessed. Spermatozoa motility and concentration were estimated with the computer-assisted semen analysis system. The semen samples were frozen using a Tris extender containing either 15% egg yolk or 5% gum arabic. For antioxidant-supplemented extenders, cysteine or ascorbic acid was dissolved at concentrations of 0.10, 0.50 or 1.0 mM in egg yolk or gum arabic extender. The semen from each ejaculate of each ram were resuspended with a specific extender with glycerol (5%); the final volume after dilution was 1 mL semen to 4 mL extender. The samples were then cooled to 4 °C for 120 min, loaded into 0.5 mL straws and frozen in liquid nitrogen for 7 days. Supplementation of gum arabic or egg yolk extenders for ram semen with antioxidants such as cysteine or ascorbic acid has beneficial effects on semen quality after cold storage or cryopreservation. However, supplementation of a 5% gum arabic extender with cysteine at 0.5 or 1 mM concentration or ascorbic acid at 0.5 mM concentration improved the quality of spermatozoa postcryopreservation. It could be concluded that gum arabic is a good alternative for egg yolk in Noemi ram semen extenders. Antioxidants are necessary to support the addition of gum arabic to the extender to help the ram spermatozoa to survive freezing–thawing and oxidative stresses. Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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10 pages, 1149 KiB

Open AccessArticle

Using Dextran Instead of Egg Yolk in Extender for Cryopreservation of Spermatozoa of Dogs of Different Ages

by Taisiia Yurchuk, Olena Pavlovich and Maryna Petrushko

Animals 2022, 12(24), 3480; https://doi.org/10.3390/ani12243480 - 9 Dec 2022

Cited by 1 | Viewed by 1776

Abstract

Egg yolk is a very common supplement of extenders aimed to protect sperm from cryoinjury, but due to their biological risks and difficulties with media standardization, there is a search for alternative. In addition, sperm cryoresistance can be affected by the initial decrease [...] Read more.

Egg yolk is a very common supplement of extenders aimed to protect sperm from cryoinjury, but due to their biological risks and difficulties with media standardization, there is a search for alternative. In addition, sperm cryoresistance can be affected by the initial decrease of their functional characteristics caused by age. The aim of this work was to evaluate the efficiency of using dextran (molecular weight 500 kDa) in the extenders instead of egg yolk for the cryopreservation of spermatozoa of dogs (Chinese Crested breed) of different ages. The obtained ejaculates were divided into three groups depending on the animal’s age: 1–3, 4–6 and 7–10 years old. Sperm was cryopreserved by using 7% glycerol and 20% egg yolk, or 20% dextran. The cryoresistance of spermatozoa of the oldest age category was dramatically decreased, which was manifested in their morphology, motility, and DNA fragmentation rate. There were no differences between the cryoprotectant effect of the dextran-based extender on spermatozoa and the egg yolk-based extender in all age categories of dogs. However, given the benefits of dextran-containing media, its use for the cryopreservation of canine spermatozoa has potential benefits that need to be confirmed by sperm fertilization outcomes. Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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16 pages, 1095 KiB

Open AccessArticle

Effect of a Novel Hydroxybenzoic Acid Based Mitochondria Directed Antioxidant Molecule on Bovine Sperm Function and Embryo Production

by João Campos Santos, Carla Cruz Marques, Maria Conceição Baptista, Jorge Pimenta, José Teixeira, Liliana Montezinho, Fernando Cagide, Fernanda Borges, Paulo J. Oliveira and Rosa M. L. N. Pereira

Animals 2022, 12(7), 804; https://doi.org/10.3390/ani12070804 - 22 Mar 2022

Cited by 4 | Viewed by 2927

Abstract

Sperm cells are particularly vulnerable to reactive oxygen species (ROS), impairing their fertilizing ability. Our objective was to study the effect of a novel mitochondrial-directed antioxidant, AntiOxBEN2, on bovine sperm function. This antioxidant was added to the semen capacitation medium (CAP), during the [...] Read more.

Sperm cells are particularly vulnerable to reactive oxygen species (ROS), impairing their fertilizing ability. Our objective was to study the effect of a novel mitochondrial-directed antioxidant, AntiOxBEN2, on bovine sperm function. This antioxidant was added to the semen capacitation medium (CAP), during the swim-up process, and to the fertilization medium (FERT) during the co-incubation of matured oocytes and capacitated spermatozoa, in concentrations of 0 (control), 1, and 10 µM. After the swim-up, sperm motility (CASA and visual analysis), vitality (eosin-nigrosin), mitochondrial membrane potential (JC1), intracellular ROS, adenosine triphosphate (ATP) levels, and basal metabolism (Seahorse Xfe96) were evaluated. Embryo development and quality were also assessed. Higher cleavage rates were obtained when 1 µM AntiOxBEN2 were added to CAP and FERT media (compared to control, p < 0.04). A positive effect of AntiOxBEN2 on intracellular ROS reduction (p = 0.01), on the increment of mitochondrial membrane potential (p ≤ 0.003) and, consequently, on the sperm quality was identified. However, the highest dose impaired progressive motility, ATP production, and the number of produced embryos. The results demonstrate a beneficial effect of AntiOxBEN2 (1 µM) on sperm capacitation and fertilization processes, thus improving embryonic development. This may constitute a putative novel therapeutic strategy to improve the outcomes of assisted reproductive techniques (ART). Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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15 pages, 1776 KiB

Open AccessArticle

Early Pregnancy in Jennies in the Caribbean: Corpus Luteum Development and Progesterone Production, Uterine and Embryo Dynamics, Conceptus Growth and Maturation

by Lorenzo G. T. M. Segabinazzi, Brandy N. Roberts, Erik W. Peterson, Rachael Ambrosia, Don Bergfelt, Juan Samper, Hilari French and Robert O. Gilbert

Animals 2022, 12(2), 127; https://doi.org/10.3390/ani12020127 - 6 Jan 2022

Cited by 2 | Viewed by 5742

Abstract

We aimed to characterize early embryo development and changes in corpus luteum (CL) development and progesterone profile in pregnant vs. non-pregnant jennies. Eight jennies were enrolled in the study. In the first two cycles, the jennies were monitored by transrectal ultrasonography and had [...] Read more.

We aimed to characterize early embryo development and changes in corpus luteum (CL) development and progesterone profile in pregnant vs. non-pregnant jennies. Eight jennies were enrolled in the study. In the first two cycles, the jennies were monitored by transrectal ultrasonography and had blood harvested for hormone profile assay. In the third cycle, jennies were bred by a jack of proven fertility. Jennies were then monitored and sampled for up to 30 days of pregnancy. Data were evaluated by random-effects multiple linear regression, and correlations were expressed as Pearson’s correlation coefficient. Progesterone concentration rose rapidly from ovulation (D0) until D7, plateaued until D12–14, then precipitously declined between D14 and 15, remaining low until the next ovulation in non-pregnant cycles. In the pregnant jennies, the progesterone concentration rose to maximal concentrations on D7–11, being higher at this stage than in non-pregnant cycles, then declined gradually up to D30. In all cycles, the volume of the CL increased steadily until D6, when it plateaued in pregnant jennies. For non-pregnant jennies, CL volume decreased slowly from D6 to D11 and then had a faster drop. Uterine tone increased following ovulation, becoming turgid around the day of embryo fixation (D15.0 ± 0.9). An embryonic vesicle (EV) was first detected on D9.3 ± 0.5 (2.4 ± 0.5 mm). The EV remained spherical until D18.6 ± 1.4. The embryo proper was first detected ventrally in the vesicle on D20.8 ± 1.1 and the embryonic heartbeat by D22.0 ± 0.9. The allantoic sac was identified at D24.0 ± 0.9, and at D30, the allantoic sac filled the ventral half of the EV. This study provides evidence that higher cumulative concentrations of progesterone are correlated to size of the EV, and there were changes in the luteal dynamics and progesterone profiles in pregnant vs. non-pregnant jennies. Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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9 pages, 1848 KiB

Open AccessArticle

Changes in Expression of Complement Components in the Ovine Spleen during Early Pregnancy

by Ling Yang, Luyu Wang, Jiaxuan Wu, Haichao Wang, Gengxin Yang and Leying Zhang

Animals 2021, 11(11), 3183; https://doi.org/10.3390/ani11113183 - 8 Nov 2021

Cited by 2 | Viewed by 2082

Abstract

During early gestation in humans, complement regulation is essential for normal fetal growth. It is supposed that a complement pathway participates in maternal splenic immune regulation at the early stage of gestation in ewes. The aim of this study was to analyze the [...] Read more.

During early gestation in humans, complement regulation is essential for normal fetal growth. It is supposed that a complement pathway participates in maternal splenic immune regulation at the early stage of gestation in ewes. The aim of this study was to analyze the effects of early pregnancy on the expression of complement components in the maternal spleen of ewes. In this study, ovine spleens were sampled on day 16 of nonpregnancy, and days 13, 16 and 25 of gestation. RT-qPCR, Western blot and immunohistochemical analysis were used to detect the changes in expression of complement components in the ovine maternal spleens. Our results reveal that C1q was upregulated during early gestation, C1r, C1s, C2, C3 and C5b increased at day 25 of gestation and C4a and C9 peaked at days 13 and 16 of gestation. In addition, C3 protein was located in the capsule, trabeculae and splenic cords. In conclusion, our results show for the first time that there was modification in the expression of complement components in the ovine spleen at the early stage of gestation, and complement pathways may participate in modulating splenic immune responses at the early stage of gestation. Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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19 pages, 1241 KiB

Open AccessArticle

Anti-Aging Effect of Urolithin A on Bovine Oocytes In Vitro

by Élisa Fonseca, Carla Cruz Marques, Jorge Pimenta, Joana Jorge, Maria Conceição Baptista, Ana Cristina Gonçalves and Rosa M. L. N. Pereira

Animals 2021, 11(7), 2048; https://doi.org/10.3390/ani11072048 - 9 Jul 2021

Cited by 7 | Viewed by 4878

Abstract

Oxidative stress and mitochondrial dysfunction have been associated with the age-related decline of oocyte quality and strategies for their prevention are currently quested. Urolithin A (UA) is a natural metabolite with pro-apoptotic and antioxidant effects, capable of preventing the accumulation of dysfunctional mitochondria [...] Read more.

Oxidative stress and mitochondrial dysfunction have been associated with the age-related decline of oocyte quality and strategies for their prevention are currently quested. Urolithin A (UA) is a natural metabolite with pro-apoptotic and antioxidant effects, capable of preventing the accumulation of dysfunctional mitochondria in different aged cells. UA has never been tested in bovine oocytes. Our aim was to study the effect of UA on the developmental potential of cumulus-oocyte-complexes (COCs) and granulosa cells’ (GCs) expression of important genes related to reproductive competence. Nuclear maturation progression, mitochondrial membrane potential (MMP) and developmental competence of physiologically mature (22 h) and in vitro aged oocytes (30 h of IVM) obtained from prepubertal and adult females, either supplemented with UA or not were assessed. Additionally, the amount of mRNA of several genes (NFE2L2, NQO1, and mt-DN5) and the number of mt-ND5 DNA copies were quantified in cultured GCs from prepubertal and adult females, either supplemented with UA or not. Our study confirmed the harmful effect of oocyte aging on the nuclear maturation progression, MMP, developmental competence and gene expression levels. UA treatment during in vitro maturation enhanced (p < 0.05) the maturation rate and subsequent developmental capacity of aged oocytes. A positive effect (p < 0.05) of UA on physiological maturation, MMP and embryonic development was also identified. UA also interfered on the expression profile of NFE2L2 and NQO1 genes in GCs cultures. Our findings demonstrate that UA supplementation is an effective way to prevent oocyte aging and improves the subsequent bovine embryonic development. Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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Review

Jump to: Research

11 pages, 699 KiB

Open AccessReview

Impact of Heat Stress on Bovine Sperm Quality and Competence

by Luís Capela, Inês Leites, Ricardo Romão, Luís Lopes-da-Costa and Rosa M. Lino Neto Pereira

Animals 2022, 12(8), 975; https://doi.org/10.3390/ani12080975 - 9 Apr 2022

Cited by 22 | Viewed by 5817

Abstract

Global warming has negatively influenced animal production performance, in addition to animal well-being and welfare, consequently impairing the economic sustainability of the livestock industry. Heat stress impact on male fertility is complex and multifactorial, with the fertilizing ability of spermatozoa affected by several [...] Read more.

Global warming has negatively influenced animal production performance, in addition to animal well-being and welfare, consequently impairing the economic sustainability of the livestock industry. Heat stress impact on male fertility is complex and multifactorial, with the fertilizing ability of spermatozoa affected by several pathways. Among the most significative changes are the increase in and accumulation of reactive oxygen species (ROS) causing lipid peroxidation and motility impairment. The exposure of DNA during the cell division of spermatogenesis makes it vulnerable to both ROS and apoptotic enzymes, while the subsequent post-meiotic DNA condensation makes restoration impossible, harming later embryonic development. Mitochondria are also susceptible to the loss of membrane potential and electron leakage during oxidative phosphorylation, lowering their energy production capacity under heat stress. Although cells are equipped with defense mechanisms against heat stress, heat insults that are too intense lead to cell death. Heat shock proteins (HSP) belong to a thermostable and stress-induced protein family, which eliminate protein clusters and are essential to proteostasis under heat stress. This review focuses on effects of heat stress on sperm quality and on the mechanisms leading to defective sperm under heat stress. Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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18 pages, 1567 KiB

Open AccessReview

Factors Affecting the Survival of Ram Spermatozoa during Liquid Storage and Options for Improvement

by Natalie Rizkallah, Caitlin G. Chambers, Simon P. de Graaf and Jessica P. Rickard

Animals 2022, 12(3), 244; https://doi.org/10.3390/ani12030244 - 20 Jan 2022

Cited by 29 | Viewed by 5185

Abstract

Semen preservation is an essential component of reproductive technologies, as it promotes genetic gain and long-distance semen transport and multiplies the number of ewes able to be inseminated per single ejaculate. However, the reduced temperature during cold storage at 5 or 15 °C [...] Read more.

Semen preservation is an essential component of reproductive technologies, as it promotes genetic gain and long-distance semen transport and multiplies the number of ewes able to be inseminated per single ejaculate. However, the reduced temperature during cold storage at 5 or 15 °C inflicts sub-lethal damage to spermatozoa, compromising sperm quality and the success of artificial breeding. New and emerging research in various species has reported the advantages of storing spermatozoa at higher temperatures, such as 23 °C; however, this topic has not been thoroughly investigated for ram spermatozoa. Despite the success of storing spermatozoa at 23 °C, sperm quality can be compromised by the damaging effects of lipid peroxidation, more commonly when metabolism is left unaltered during 23 °C storage. Additionally, given the biosafety concern surrounding the international transport of egg-yolk-containing extenders, further investigation is critical to assess the preservation ability of synthetic extenders and whether pro-survival factors could be supplemented to maximise sperm survival during storage at 23 °C. Full article

(This article belongs to the Special Issue Advances in Animal Fertility Preservation)

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