Impact of Oxidative Stress and Antioxidants on Semen Quality in Dogs

Studies showed a beneficial effect of supplementation with selenium (Se) and vitamin E on semen quality. The aim of the study was to investigate the effect of Se and vitamin E supplementation on the antioxidant status of spermatozoa and semen quality in dogs with lowered fertility. Ten dogs were supplemented daily with Se (6μg/kg organic Se yeast) and vitamin E (5mg/kg) per os for 60days. Control group consisted of 10 males without the supplementation. Semen was collected on day 0, 30, 60 and 90. Sperm quality parameters were evaluated using CASA and a microscope. Concentrations of Se and vitamin E in blood as well as glutathione peroxidase (GSH-Px) activity and total antioxidant capacity (TAC) in the spermatozoa were determined. After 60days of supplementation the concentration of spermatozoa, the majority of motility indicators and the percentage of normal morphology and live spermatozoa increased significantly (p<.05). An increase (p<.05) in concentration of Se and vitamin E in blood and GSH-Px-activity and TAC in the spermatozoa was detected. The study results indicate that Se and vitamin E supplementation for 60days enhances the antioxidant status of spermatozoa and improves the quality of the semen in dogs with lowered fertility.

Effect of clavulanic acid-potentiated amoxycillin on semen quality in dogs

Oxidative stress owing to an imbalance between reactive oxygen species and antioxidants, such as coenzyme Q10 (CoQ10), is a major contributor to maleinfertility. We investigated the effects of the reduced form of CoQ10 (ubiquinol) supplementation on semen quality in dogs with poor semen quality. Three dogsreceived 100 mg of ubiquinol orally once daily for 12 weeks. Semen quality, serum testosterone, and seminal plasma superoxide dismutase (SOD) activity wereexamined at 2-week intervals from 2 weeks before ubiquinol supplementation to 4 weeks after the treatment. Ubiquinol improved sperm motility, reducedmorphologically abnormal sperm, and increased seminal plasma SOD activity; however, it had no effect on testosterone level, semen volume, and sperm number.Ubiquinol supplementation could be used as a non-endocrine therapy for infertile dogs.

Griseofulvin is used to treat dermatomycosis in many species and causes oligospermia in supra-pharmacological doses. The aim of the present study was to evaluate the effect of Griseofulvin administered at therapeutic doses upon semen quality in dogs. Four dogs were treated with Griseofulvin (25 mg/kg per day) for 30 days. Semen collections and analyses were performed before, during and for 100 days after treatment for the Griseofulvin group and 10 untreated control dogs. Semen analyses included mean percentage of forward progressively motile sperm, total sperm output, normal live sperm and normal dead sperm. There was no significant difference between control and treated dogs for each of the semen quality parameters. Therapeutic dosage of Griseofulvin had no deleterious effect upon semen quality in dogs, although this does not preclude potential embryotoxic and teratogenic effects.

Simple SummaryThe wide use of artificial insemination in dogs justifies the development of new strategies to prevent the reduction of fertilizing ability of stored semen. In recent years, the use of plant antioxidant supplementation has become increasingly popular. Maca (Lepidium meyenii) is an Andean edible root with antioxidant properties. The effectiveness of the oral supplementation of Maca in improving fresh semen quality and quantity and cooling or freezing ability has already been reported. This is the first in vitro study on the effects of aqueous extract of Maca on canine spermatozoa. The addition of low concentrations of aqueous extract of Maca to the canine chilled extender had positive effects only until 24 h of storage, increasing hyperactivation of sperm cells and preserving DNA integrity of spermatozoa in short-term storage. Meanwhile, a high concentration of Maca had an immediately deleterious effect on semen quality. Antioxidant supplementation has been proposed as a new strategy to improve the long-term preservation of semen. The aim of this study was to evaluate the effect of Maca supplementation of semen extender on quality-related canine semen parameters during cooling. Ejaculates from nine dogs were cooled for 7 days in the absence (control group) or in the presence of 10, 20 and 50 μL/mL of an aqueous extract of Maca. Sperm were evaluated for sperm viability, motility, DNA fragmentation and lipid peroxidation after 3 h, 24 h, 4 days and 7 days of storage. The addition of 10 μL/mL of Maca preserved sperm DNA and plasma membrane integrity at 3 h and increased sperm curvilinear velocity after 24 h. Treatment with 20 and 50 μL/mL of Maca increased the percentage of hyperactivated sperm after 3 h. Moreover, semen treated with 20 μL/mL of Maca decreased lipid peroxidation at 24 h. A significant reduction of sperm DNA and plasma membrane integrity as well as of kinetics parameters between 3 and 24 h of refrigerated storage with the higher concentration tested was observed. Although Maca was not able to protect canine semen with extended refrigeration storage time, it increased hyperactivation and preserved DNA integrity in short-term storage.

Benign prostatic hyperplasia (BPH) may alter prostatic fluid biochemical composition causing reduced fertility. Osaterone acetate (OA) is an androgen receptor antagonist marketed for treatment of canine BPH. Little information exists on effects of OA administration on biochemical composition of canine prostatic fluid and its role on fertility. The aim of this research was to study biochemical composition of prostatic fluid and its role on semen quality in dogs with BPH undergoing treatment with OA. Eight intact, 5-11-year-old dogs with benign prostatic hyperplasia were treated orally with OA at a dose of 0.25-0.5mg/kg once daily for seven days. Prostatic volume, semen evaluation and a biochemical analysis of prostatic fluid were performed on the day before treatment (D0), D60, D120, D180 and D240. A significant reduction (57% and 61%) of prostatic volume was observed at D60 and D120, respectively, and a significant reduction (20%) of normal spermatozoa was observed at D60 coincident with a significant increase of sperm tail defects, which disappeared during the course of the treatment. Prostatic fluid composition did not vary during the OA treatment except for zinc (Zn2+ ) with a significant increase at D120 and D180 correlated with the return to normal sperm values. In conclusion, canine Zn2+ prostatic fluid concentrations decrease during development of BPH and return to normal during treatment with OA. Zn2+ is an important electrolyte for semen quality, suggesting that oral Zn2+ supplementation might be considered a treatment to improve semen quality.

The semen evaluation techniques used in most commercial artificial insemination centers, which includes sperm motility and morphology measurements, provides a very conservative estimate of the relative fertility of individual boars. As well, differences in relative boar fertility are masked by the widespread use of pooled semen for commercial artificial insemination (AI) in many countries. Furthermore, the relatively high sperm numbers used in commercial AI practice usually compensate for reduced fertility, as can be seen in some boars when lower numbers of sperm are used for AI. The increased efficiency of pork production should involve enhanced use of boars with strong reproductive efficiency and the highest genetic merit for important production traits. Given that the current measures of semen quality are not always indicative of fertility and reproductive performance in boars, accurate and predictive genetic and protein markers are still needed. Recently, significant efforts have been made to identify reliable markers that allow for the identification and exclusion of sires with reduced reproductive efficiency. This paper reviews the current status of proteomic and genomic markers of fertility in boars in relation to other livestock species.

Sperm quality and selected biochemical parameters of seminal fluid in dogs with benign prostatic hyperplasia

Effect of seminal plasma vesicular structures in canine frozen-thawed semen

Thirty clinically healthy dogs with poor semen quality were used in the study. Fifteen dogs were supplemented daily with selenium (0.6 mg/kg organic selenium from yeast) and vitamin E (5 mg/kg) per os for 60 d. The control group (15 dogs) was not supplemented. Semen was collected from all dogs by manual manipulation on days 0, 30, 60, and 90. The sperm concentration and motility parameters were evaluated with a Hamilton Thorne sperm analyser, version IVOS 12.3. For the assessment of sperm morphology, Diff-Quik stain was used. The percentage of live and dead spermatozoa was estimated on dried smears stained with eosin-nigrosin. The concentration of spermatozoa, most motility parameters determined (PMOT, VSL, VCL, ALH, BCF, RAPID, MEDIUM, SLOW, and STATIC), and the percentage of spermatozoa morphologically normal and live increased significantly (P &lt; 0.05) after 60 d of supplementation. In the control group, there were no changes in motility parameters while the concentration and total sperm count decreased over the duration of the study. In conclusion, supplementation with selenium and vitamin E for 60 d can improve the quality of semen in dogs with lowered fertility.

Simple SummaryIn rabbit farms, artificial insemination is usually accepted using semen preserved around 18 °C. However, the use of cryopreserved rabbit semen is limited, due to excess oxidative stress and produce sperm dysfunction. The advancements in nanotechnology tools have allowed molecular-based targeting of cells through effective, safe, and biocompatible magnetic nanoparticles with promising potentials in reproductive sciences. In these regards, the current work aimed to explore the potential role if the effect of curcumin nanoparticles supplementation in semen extender on post/thawed rabbit sperm quality. Results revealed that the CUNPs (1.5 µg/mL) showed superior enhancements impacts for the post-thawing sperm motion and redox status, as well as a significant reduction in apoptotic and necrotic sperm cells. This confirmed the constructive application of nanoparticle to enhance the cryopreserved rabbit’s sperm function.The cryopreservation process adversely affects sperm function and quality traits, causing some changes at biochemical and structural levels, due to mechanical, thermal, osmotic, and oxidative damage. Supplementation with curcumin nanoparticles could prevent and even revert this effect and could enhance the post/thawed sperm quality in the rabbit. The study amid to explore the effect of curcumin (CU) and curcumin nanoparticles (CUNPs) supplementation in semen extender on post/thawed rabbit sperm quality. Twelve fertile, healthy rabbit bucks were included, and the ejaculates were collected using artificial vaginas. Rabbit pooled semen was cryopreserved in tris-yolk fructose (TYF) extender without any supplement (control group) or extender supplemented with CU at levels of 0.5, 1 or 1.5 µg/mL (CU0.5, CU1.0, and CU1.5, respectively) or CUNPs at levels of 0.5, 1, 1.5 (CUNPs0.5, CUNPs1.0, and CUNPs1.5, respectively) and was packed in straws (0.25 mL) and stored in liquid nitrogen (−196 °C). Results revealed that CUNPs1.5 had a positive influence (p < 0.05) on post-thawing sperm progressive motility, viability, and membrane integrity as compared with the other groups. Percentages of dead sperm, abnormalities, early apoptotic, apoptotic, and necrotic sperm cells reduced (p < 0.05) in CUNPs1.5 as compared to other treatments. Using 1.5 µg/mL of CUNPs significantly improved total antioxidant capacity (TAC), GPx, while MDA and POC reduced (p < 0.05) in CU1.5 in comparison with other groups. SOD values were enhanced (p < 0.05) in CUNPs1.0 and CUNPs1.5 in relation with other treatments. Conclusively, the addition of curcumin and its nanoparticles to the extender can improve the post-thawed quality of rabbit sperm via redox signaling and reduce the apoptosis process.

Nurture vs nature: how can we optimize sperm quality?

Simple SummaryCryopreservation of semen is getting easier, however, fertilizing results after insemination with frozen-thawed semen is still not constant in canine species depending on the breed and could be still improved. In this study, we decided to modulate the mitochondrial activity through the addition of metformin in semen extender to increase germ cells’ quality. Metformin presented the absence of toxicity and an improvement in sperm motility after thawing, as well as an increase in the expression of several molecular markers associated with quality. In addition, the oxidative stress and DNA damage were reduced in semen frozen in the presence of metformin. Overall, these data suggest that metformin added in canine semen extender has beneficial effects on canine semen quality and could be associated with different components such as vitamins, to enhance the antioxidants status.Sperm cryopreservation is an assisted reproductive technique routinely used in canine species for genetic conservation. However, during cryopreservation, the DNA damages are still elevated, limiting the fertilization rate. The present study was conducted to evaluate whether supplementation of canine semen extender with a molecule limiting the metabolic activities can improve the quality of frozen-thawed canine spermatozoa. We used metformin, known to limit the mitochondrial respiratory and limit the oxidative stress. Before and during the freezing procedure, metformin (50 µM and 500 µM) has been added to the extender. After thawing, sperm exposed to metformin conserved the same viability without alteration in the membrane integrity or acrosome reaction. Interestingly, 50 µM metformin improved the sperm motility in comparison to the control, subsequently increasing mitochondrial activity and NAD+ content. In addition, the oxidative stress level was reduced in sperm treated with metformin improving the sperm quality as measured by a different molecular marker. In conclusion, we have shown that metformin is able to improve the quality of frozen-thawed dog semen when it is used during the cryopreservative procedure.

The purpose of this study was to determine the impact of an antimicrobial peptide, BiF2_5K7K, on semen quality and bacterial contamination in boar semen doses used for artificial insemination. A key factor affecting semen quality and farm production is bacterial contamination in semen doses. Using antibiotics in a semen extender seems to be the best solution for minimizing bacterial growth during semen preservation. However, concern regarding antibiotic-resistant microorganisms has grown globally. As a result, antimicrobial peptides have emerged as interesting alternative antimicrobial agents to replace the current antibiotics used in semen extenders. BiF2_5K7K is an antimicrobial peptide that can inhibit Gram-negative and Gram-positive bacteria isolated from boar semen and sow vaginal discharge. In this study, ten fresh boar semen samples were collected and diluted with one of two types of semen extender: with (positive control) or without (negative control) an antibiotic (i.e., gentamicin). The semen extender without an antibiotic contained antimicrobial peptide BiF2_5K7K at different concentrations (15.625, 31.25, 62.5, and 125 µg/mL). The samples were stored at 18 °C until use. Semen quality parameters were assessed on days 0, 1, 3, and 5, and the total bacterial count was also evaluated at 0, 24, 36, 48, and 72 h after storage. A fertility test on a pig farm was also performed via sow insemination with a commercial extender plus BiF2_5K7K at a concentration of 31.25 µg/mL. No significant difference was found in terms of semen quality on days 0 or 1. On days 3 and 5, the total motility, progressive motility, and viability remained normal in the 15.625 and 31.25 µg/mL groups. However, the sperm parameters decreased starting on day 3 for the 125 µg/mL group and on day 5 for the 62.5 µg/mL group. For total bacterial count at 0, 24, 36, 48, and 72 h, the lowest bacterial count was found in the positive control group, and the highest bacterial count was found in the negative control group compared with the other groups. Comparing antimicrobial peptide groups from 0 to 48 h, the lowest bacterial count was found in the 125 µg/mL group, and the highest bacterial count was found in the 15.625 µg/mL group. For the fertility test, artificial insemination was conducted by using a commercial extender plus BiF2_5K7K at a concentration of 31.25 µg/mL. The results showed a superior pregnancy rate, farrowing rate, and total number of piglets born compared with artificial insemination conducted using a commercial extender plus antibiotic. In conclusion, BiF2_5K7K can inhibit bacterial growth in extended boar semen for 24 h, and thereafter, the bacterial count slightly increases. However, the increase in the number of bacterial counts from days 0 to 3 had no negative effect on sperm quality in the positive control, 15.625, or 31.25 µg/mL groups. This indicates that BiF2_5K7K might be an antimicrobial peptide candidate with potential for use as an alternative antimicrobial agent to replace the conventional antibiotic used in boar semen extenders.

Semen evaluation and fertility assessment in a purebred dog breeding facility