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Pharmacological Activity of Honeybush (Cyclopia intermedia) in Boar Spermatozoa during Semen Storage and under Oxidative Stress.

ABSTRACT: In recent decades, an increasing number of ethnopharmacological studies have been dedicated to medicinal plants from South African fynbos. Among these plants, honeybush (Cyclopia spp.) has become a popular tea, mainly due to its healthy properties and caffeine-free status. The antioxidant, antimutagenic, and antimicrobial properties of this plant have been reported in several cell types, but its effects on reproductive function are still unknown. Here, we assessed the effects of honeybush (Cyclopia intermedia) on boar sperm parameters under induced oxidative stress (Fe2+/ascorbate) and during five days of semen storage at 17 °C without oxidative stress. In both experiments, four concentrations (200, 50, 12.5, and 3.125 µg/mL) of fermented honeybush were tested. Our results show that honeybush enhances sperm parameters, and no toxic effects were observed at any of the tested extract concentrations. Interestingly, honeybush (12.5 µg/mL) improved the sperm motility and kinetic parameters, preserved the plasma membrane integrity, and reduced the lipid peroxidation in the samples exposed to Fe2+/ascorbate (p < 0.05). In the stored samples, positive effects of honeybush on sperm parameters (motility, kinetics, acrosome, and mitochondria) were observed from 48 h until 120 h of semen storage (p < 0.05). Our results clearly show the protective effects of honeybush on sperm samples, thus promoting its use as a natural source of antioxidants for boar semen.

SUBMITTER: Ros-Santaella JL

PROVIDER: S-EPMC7142635 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Pharmacological Activity of Honeybush (<i>Cyclopia intermedia</i>) in Boar Spermatozoa during Semen Storage and under Oxidative Stress.

Ros-Santaella José Luis JL Kadlec Martin M Pintus Eliana E

Animals : an open access journal from MDPI 20200310 3

In recent decades, an increasing number of ethnopharmacological studies have been dedicated to medicinal plants from South African fynbos. Among these plants, honeybush (<i>Cyclopia</i> spp.) has become a popular tea, mainly due to its healthy properties and caffeine-free status. The antioxidant, antimutagenic, and antimicrobial properties of this plant have been reported in several cell types, but its effects on reproductive function are still unknown. Here, we assessed the effects of honeybush ...[more]

PMID: 32164369

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Project description:Background:The current cultivation and plant breeding of Honeybush tea (produced from members of Cyclopia Vent.) do not consider the genetic diversity nor structuring of wild populations. Thus, wild populations may be at risk of genetic contamination if cultivated plants are grown in the same landscape. Here, we investigate the spatial distribution of genetic diversity within Cyclopia intermedia E. Mey.-this species is widespread and endemic in the Cape Floristic Region (CFR) and used in the production of Honeybush tea. Methods:We applied High Resolution Melt analysis (HRM), with confirmation Sanger sequencing, to screen two non-coding chloroplast DNA regions (two fragments from the atpI-aptH intergenic spacer and one from the ndhA intron) in wild C. intermedia populations. A total of 156 individuals from 17 populations were analyzed for phylogeographic structuring. Statistical tests included analyses of molecular variance and isolation-by-distance, while relationships among haplotypes were ascertained using a statistical parsimony network. Results:Populations were found to exhibit high levels of genetic structuring, with 62.8% of genetic variation partitioned within mountain ranges. An additional 9% of genetic variation was located amongst populations within mountains, suggesting limited seed exchange among neighboring populations. Despite this phylogeographic structuring, no isolation-by-distance was detected (p > 0.05) as nucleotide variation among haplotypes did not increase linearly with geographic distance; this is not surprising given that the configuration of mountain ranges dictates available habitats and, we assume, seed dispersal kernels. Conclusions:Our findings support concerns that the unmonitored redistribution of Cyclopia genetic material may pose a threat to the genetic diversity of wild populations, and ultimately the genetic resources within the species. We argue that 'duty of care' principles be used when cultivating Honeybush and that seed should not be translocated outside of the mountain range of origin. Secondarily, given the genetic uniqueness of wild populations, cultivated populations should occur at distance from wild populations that is sufficient to prevent unintended gene flow; however, further research is needed to assess gene flow within mountain ranges.

Project description:Genetic markers have been used to assess the freezability of semen. With the advancement in molecular genetic techniques, it is possible to assess the relationships between sperm functions and gene polymorphisms. In this study, variant calling analysis of RNA-Seq datasets was used to identify single nucleotide polymorphisms (SNPs) in boar spermatozoa and to explore the associations between SNPs and post-thaw semen quality. Assessment of post-thaw sperm quality characteristics showed that 21 boars were considered as having good semen freezability (GSF), while 19 boars were classified as having poor semen freezability (PSF). Variant calling demonstrated that most of the polymorphisms (67%) detected in boar spermatozoa were at the 3'-untranslated regions (3'-UTRs). Analysis of SNP abundance in various functional gene categories showed that gene ontology (GO) terms were related to response to stress, motility, metabolism, reproduction, and embryo development. Genomic DNA was isolated from sperm samples of 40 boars. Forty SNPs were selected and genotyped, and several SNPs were significantly associated with motility and membrane integrity of frozen-thawed (FT) spermatozoa. Polymorphism in SCLT1 gene was associated with significantly higher motility and plasma membrane integrity of FT spermatozoa from boars of the GSF group compared with those of the PSF group. Likewise, polymorphisms in MAP3K20, MS4A2, and ROBO1 genes were significantly associated with reduced cryo-induced lipid peroxidation and DNA damage of FT spermatozoa from boars of the GSF group. Candidate genes with significant SNP associations, including APPL1, PLBD1, FBXO16, EML5, RAB3C, OXSR1, PRICKLE1, and MAP3K20 genes, represent potential markers for post-thaw semen quality, and they might be relevant for future improvement in the selection procedure of boars for cryopreservation. The findings of this study provide evidence indicating that polymorphisms in genes expressed in spermatozoa could be considered as factors associated with post-thaw semen quality.

Project description:Artificial insemination (AI) is the preferred method for reproduction in the majority of the intensive pig production systems Worldwide. To this end, fresh extended ready-to-use semen doses are either purchased from AI-centres or produced by boars kept on-farm. For profitable semen production, it is necessary to obtain a maximum amount of high quality semen from each boar. This paper reviews current knowledge on factors that may affect semen quality by influencing the boar or the semen during processing. Genetic markers could be used for early detection of boars with the highest fertility potential. Genetic selection for fast growth might jeopardize semen quality. Early detection of boars no longer fit for semen production might be possible by ultrasonography of the testes. Seasonal variation in sperm quality could be associated with changes in photoperiod and heat stress during summer. Comfortable housing, with appropiate bedding material to avoid locomotion problems is essential. In some areas, cooling systems may be necessary to avoid heat stress. The sperm quality can be manipulated by feeding strategies aiming, for instance, to increase sperm resistance to oxidative stress and extend storage duration. High collection frequency will negatively influence sperm quality. Also, if collection is not hygienically performed it will result in bacterial contamination of the semen doses. The concern over bacterial contamination has risen not only because of its negative effect on semen quality but also due to the detection of antimicrobial resistance in isolates from extended semen. Moreover, bacterial and viral pathogens must be monitored because they affect semen production and quality and constitute a risk of herd infection. During processing, boar sperm are submitted to many stress factors that can cause oxidative stress and capacitation-like changes potentially reducing their fertility potential. Dilution rate or dilution temperature affects the quality of the semen doses. Some packaging might preserve semen better than others and some plastic components might be toxic for sperm. Standard operation procedures and quality assurance systems in AI centres are needed.

Dataset Information

Pharmacological Activity of Honeybush (Cyclopia intermedia) in Boar Spermatozoa during Semen Storage and under Oxidative Stress.

Publications

Pharmacological Activity of Honeybush (<i>Cyclopia intermedia</i>) in Boar Spermatozoa during Semen Storage and under Oxidative Stress.

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