The Antibacterial and Antioxidant Roles of Buckwheat Honey (BH) in Liquid Preservation of Boar Semen.
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ABSTRACT:
SUBMITTER: Lan Q
PROVIDER: S-EPMC8192209 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Although the bacterial composition of boar ejaculate has been extensively studied, the bacterial composition of extended boar semen is often overlooked, despite the potential risks these microorganisms may pose to the long-term preservation of extended boar semen at 15-17°C. In this study, we characterized the bacterial community composition of extended semen and discovered that Pseudomonas spp. was the dominant flora. The dominant strains were further isolated and identified as a potential new species in the Pseudomonas fluorescens group and named GXZC strain, which had adverse effects on sperm quality and was better adapted to growth at 17°C. Antimicrobial susceptibility testing showed that the GXZC strain was resistant to all commonly used veterinary antibiotics. Whole-genome sequencing (WGS) and genome annotation revealed the large genetic structure and function [7,253,751 base pairs and 6,790 coding sequences (CDSs)]. Comparative genomic analysis with the closest type strains showed that the GXZC strain predicted more diversity of intrinsic and acquired resistance genes to multi-antimicrobial agents. Taken together, our study highlights a problem associated with the long-term storage of extended boar semen caused by a P. fluorescens group strain with unique biological characteristics. It is essential to develop a new antibacterial solution for the long-term preservation of boar semen.
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Project description:Various semen extender formulas are in use to maintain sperm longevity and quality whilst acting against bacterial contamination in liquid sperm preservation. Aminoglycosides are commonly supplemented to aid in the control of bacteria. As bacterial resistance is increasing worldwide, antimicrobial peptides (AMPs) received lively interest as alternatives to overcome multi-drug resistant bacteria. We investigated, whether synthetic cationic AMPs might be a suitable alternative for conventional antibiotics in liquid boar sperm preservation. The antibacterial activity of two cyclic AMPs (c-WWW, c-WFW) and a helical magainin II amide analog (MK5E) was studied in vitro against two Gram-positive and eleven Gram-negative bacteria. Isolates included ATCC reference strains, multi-resistant E. coli and bacteria cultured from boar semen. Using broth microdilution, minimum inhibitory concentrations were determined for all AMPs. All AMPs revealed activity towards the majority of bacteria but not against Proteus spp. (all AMPs) and Staphylococcus aureus ATCC 29213 (MK5E). We could also demonstrate that c-WWW and c-WFW were effective against bacterial growth in liquid preserved boar semen in situ, especially when combined with a small amount of gentamicin. Our results suggest that albeit not offering a complete alternative to traditional antibiotics, the use of AMPs offers a promising solution to decrease the use of conventional antibiotics and thereby limit the selection of multi-resistant strains.
| S-EPMC4141845 | biostudies-literature
Project description:Hypothermic storage of boar semen provides the possibility to omit antibiotics from semen extenders so long as sperm quality is maintained and bacterial growth prevented. The objective of this study was to determine an optimal cooling-rate frame for boar semen preserved at 5°C in an antibiotic-free extender. Semen from eight boars extended in AndroStar® Premium was cooled from 30°C to 5°C using seven different cooling rates, ranging initially from 0.01 to 0.36°C min-1 and reaching 5°C between 2 h and 24 h after dilution. Sperm motility, membrane integrity, membrane fluidity, mitochondrial membrane potential and the response to the capacitation stimulus bicarbonate remained at a high level for 144 h at 5°C when the semen was initially cooled in a cooling-rate frame ranging from 0.01 to 0.09°C min‑1 in the temperature zone from 30 to 25°C, followed by 0.02 to 0.06°C min-1 to 10°C and 0.01 to 0.02°C min‑1 to the final storage temperature. A cooling rate of 0.07°C min-1 in the temperature zone from 30 to 10°C led to a reduced response to bicarbonate (P < 0.01) and fast cooling to 5°C within 1 h with a cooling rate of 0.31°C min-1 resulted in lower values (P > 0.05) of all sperm parameters. In a further experiment, slow cooling with a holding time of 6 h at 22°C induced after 6 h storage a temporary increase in Escherichia coli of 0.5 × 103 to 2.4 × 103 CFU mL-1 in the sperm-free inoculated extender. Overall, the load of mesophilic bacteria in the stored semen was below 6 × 103 CFU mL-1, a level that is not regarded as critical for sperm quality. In conclusion, appropriate cooling protocols were established for the antibiotic-free storage of boar semen at 5°C, allowing the application of hypothermic preservation in research and in artificial insemination.
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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.
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Project description:Liquid preservation of boar sperm is crucial for artificial insemination application in pig production. However, time-dependent oxidative damage to sperm is one of the major challenges during the liquid preservation period. Caffeic acid phenethyl ester (CAPE) possesses excellent antioxidant properties and has potential therapeutic use in reproductive organ injury linked to oxidative stress. Adenosine monophosphate (AMP)-activated protein kinase (AMPK) involves in modulating the cellular redox state and exerts a beneficial effect on sperm preservation. In the present study, we firstly assessed different concentrations of CAPE that affect sperm quality during liquid storage to determine the appropriate addition. To further investigate whether CAPE exerts protective effects on boar sperm through modulation of AMPK activity, sperm quality parameters, antioxidant capacity, and marker protein expressions were evaluated under co-incubation with H2O2. The results showed that sperm treated with 210 μmol/L CAPE exhibited the highest motion parameters (total motility and progressive motility) and best functional integrity (mitochondrial activity, plasma membrane integrity, and acrosomal integrity). Even in the presence of H2O2, the addition of 210 μmol/L CAPE not only significantly improved sperm quality parameters, but also elevated CAT, SOD, and GSH-Px activities to enhance sperm antioxidant capacity. In addition, we found that CAPE could affect the protein activities of AMPK, phospho-AMPK α (p-AMPK), SOD, and Caspase-3 regardless of whether H2O2 is present or not. Our findings suggested that CAPE has potential application in liquid preservation of boar sperm and preliminary indicated that CAPE-induced improvement of sperm quality and antioxidant capacity should be mediated through conservation of AMPK activity. Further studies are required to illustrate the specific mechanism by which CAPE attenuates oxidative stress-mediated damages dependent on AMPK activity. Graphical Abstract The potential protective mechanism of CAPE on boar sperm. CAPE induces AMPK phosphorylation resulting in elevating the protein expression of SOD1 to eliminate ROS as well as inhibiting the protein expression of Caspase-3. Consequently, both the liquid preservation time and boar sperm quality are improved.
| S-EPMC9219730 | biostudies-literature
Project description:During cryopreservation, sperm undergoes structural and molecular changes such as ice crystal formation, DNA fragmentation, and reactive oxygen species (ROS) production, leading to decreased sperm quality after thawing. Antioxidants play a crucial role in preventing these damages, both in vivo and in vitro. One potent antioxidant is myo-inositol, known for its protective effects on sperm against ROS. This study aimed to investigate the protective effect of myo-inositol on cryopreserved boar semen. The semen was diluted, cooled, and cryopreserved using a BF5 extender. It was then divided into five groups: control and different concentrations of myo-inositol (0.5, 1, 1.5, and 2 mg/mL). The post-thaw evaluation included assessments of motility, viability, acrosome integrity, mitochondrial membrane potential (MMP), caspase activity, gene expression, ROS levels, apoptosis, and IVF with treated semen. Results showed that myo-inositol at 0.5 mg/mL improved motility, acrosome integrity, and fertilization ability. It also reduced the expression of pro-apoptotic genes and increased SMCP expression. Lower concentrations also demonstrated improved viability and reduced apoptosis and ROS levels. In conclusion, myo-inositol treatment during cryopreservation improved sperm quality, reduced apoptosis and ROS levels, and enhanced fertility rates in boar semen.
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Project description:The role of antibiotics (AB) in semen extenders as a potential contribution to the global antimicrobial resistance threat is emerging. Here, we establish an AB-free hypothermic preservation strategy for boar semen and investigate its impact on sperm function, microbial load and fertility after artificial insemination (AI). Spermatozoa (12 boars) preserved in AB-free AndroStar Premium extender at 5 °C maintained high motility, membrane integrity, and a low DNA-fragmentation index throughout 72 h storage and results did not significantly differ from controls stored at 17 °C in extender containing AB (p = 0.072). Likewise, kinetic response of spermatoza to the capacitation stimulus bicarbonate during 180 min incubation in Tyrode's medium did not differ from 17 °C-controls. In a competitive sperm oviduct binding assay, binding indices did not differ between semen stored for 72 h AB-free at 5 °C and 17 °C-controls (n = 6 boars). Bacterial load < 103 CFU/ml after 72 h was measured in 88.9% of samples stored at 5 °C AB-free compared to 97.2% in 17 °C-controls (n = 36 semen pools, 23 boars). Fertility traits of 817 females did not differ significantly between the two semen groups (p > 0.05). In conclusion, a hypothermic semen preservation strategy is presented which offers antibiotic-free storage of boar semen doses.
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Project description:A focus of research in recent years is the comparison of honey as the final product of bees with pollen and nectar of the plant from which the honey originates, as the main food source for bees. Buckwheat honey is recognized as a nutritionally valuable product, which provides a scientifically proven health benefit and is confirmed as a functional food. The quality of this type of honey is attributed to high levels of phytochemicals in buckwheat. The purpose of this study was the examination of similarity between buckwheat honey and buckwheat nectar and pollen, as well as simultaneous investigation of their chemical profiles and the origin of the honey. The phenolic profile of buckwheat pollen showed a lower number of flavonoids and phenolic acids than those of nectar and honey samples, but confirmed the presence of the most characteristic polyphenols derived from the buckwheat plant. The notable difference was found to be the presence of (epi)catechin units, its galloylated derivatives and procyanidin dimers, which were not present in honey. Honey polyphenols displayed a pronounced correlation with those of nectar, but not with those of pollen. Finally, by comparing the polyphenolic profiles of honey, nectar and pollen sharing the same geographical origin, new data could be provided for a potential assessment of the botanical origin of buckwheat honey.
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Project description:Antimicrobial resistance is an increasing challenge in semen preservation of breeding animals, especially in the porcine species. Bacteria are a natural component of semen, and their growth should be inhibited to protect sperm fertilizing capacity and the female's health. In pig breeding, where semen is routinely stored at 17°C in the liquid state, alternatives to conventional antibiotics are urgently needed. Photodynamic inactivation (PDI) of bacteria is a well-established tool in medicine and the food industry but this technology has not been widely adopted in semen preservation. The specific challenge in this setting is to selectively inactivate bacteria while maintaining sperm integrity and functionality. The aim of this study was to test the principle of PDI in liquid stored boar semen using the photosensitizer 5,10,15,20-tetrakis(N-methyl-4-pyridyl)-21H,23H-porphine (TMPyP) and a white light LED-setup. In the first step, photophysical experiments comprising singlet oxygen phosphorescence kinetics of TMPyP and determination of the photosensitizer triplet time revealed a sufficiently high production of reactive singlet oxygen in the Androstar Premium semen extender, whereas seminal plasma acted as strong quencher. In vitro experiments with extended boar semen showed that the established PDI protocol preserves sperm motility, membrane integrity, DNA integrity, and mitochondrial activity while efficiently reducing the bacteria below the detection limit. A proof-of-concept insemination study confirmed the in vivo fertility of semen after photodynamic treatment. In conclusion, using the PDI approach, an innovative tool was established that efficiently controls bacteria growth in extended boar and maintains sperm fertility. This could be a promising contribution to the One Health concept with the potential to reduce antimicrobial resistance in animal husbandry.
| S-EPMC11335528 | biostudies-literature
Project description:The aim was to investigate the effects of the cold dehulling of buckwheat seeds on their germination, total phenolic content (TPC), antioxidant activity (AA) and phenolics composition. Cold dehulling had no negative effects on germination rate and resulted in faster rootlet growth compared to hulled seeds. Although the dehulling of the seeds significantly decreased TPC and AA, the germination of dehulled seeds resulted in 1.8-fold and 1.9-fold higher TPC and AA compared to hulled seeds. Liquid chromatography coupled to mass spectrometry identified several phenolic compounds in free and bound forms. Rutin was the major compound in hulled seeds (98 µg/g dry weight), orientin and vitexin in 96-h germinated dehulled seeds (2205, 1869 µg/g dry weight, respectively). During germination, the increases in the major phenolic compounds were around two orders of magnitude, which were greater than the increases for TPC and AA. As well as orientin and vitexin, high levels of other phenolic compounds were detected for dehulled germinated seeds (e.g., isoorientin, rutin; 1402, 967 µg/g dry weight, respectively). These data show that dehulled germinated seeds of buckwheat have great potential for use in functional foods as a dietary source of phenolic compounds with health benefits.
| S-EPMC8066582 | biostudies-literature