The Lipidomics of Spermatozoa and Red Blood Cells Membrane Profile of Martina Franca Donkey: Preliminary Evaluation

Prasinou, Paraskevi; Amicis, Ippolito De; Fusaro, Isa; Bucci, Roberta; Cavallini, Damiano; Parrillo, Salvatore; Caputo, Maurizio; Gramenzi, Alessandro; Carluccio, Augusto

doi:10.3390/ani13010008

Cited by 3 publications

(3 citation statements)

References 59 publications

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“…It is important to mention the enormous variability of factors influencing sperm quality. This is the case for the fatty acid composition of the sperm membrane, which has been proven to influence sperm motility parameters and fertility in stallion semen [33]. In this regard, these authors analyzed the liposomes of the sperm membrane and the relation to the dietary regimen, highlighting the importance of polyunsaturated acid balance and demonstrating a positive correlation between oleic acid and progressive motility in equine semen.…”

Section: Discussionmentioning

confidence: 99%

Strategies to Reduce the Use of Antibiotics in Fresh and Chilled Equine Semen

Zabala,

Serres,

Montero

et al. 2024

Animals

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The study assessed the impact of four equine semen processing techniques on sperm quality and microbial load immediately post-processing and after 48 h of refrigeration. The aim was to explore the potential reduction of prophylactic antibiotic usage in semen extenders. Semen from ten adult stallions was collected and processed under a strict hygiene protocol and divided into four aliquots: Simple Centrifugation with antibiotics (SC+), Simple Centrifugation (SC−), Single-Layer Colloidal Centrifugation (CC−), and Filtration (with SpermFilter®) (F−), all in extenders without antibiotics. Sperm motility, viability, and microbial load on three culture media were assessed. No significant differences were observed in the main in the sperm quality parameters among the four protocols post-processing and at 48 h (p < 0.05 or p < 0.1). Microbial loads in Columbia 5% Sheep Blood Agar and Schaedler vitamin K1 5% Sheep Blood Agar mediums were significantly higher (p < 0.10) for raw semen than for CS+, CC−, and F− post-processing. For Sabouraud Dextrose Agar medium, the microbial load was significantly higher (p < 0.10) in raw semen compared to CS+ and F−. No significant differences (p < 0.10) were found in 48 h chilled samples. Regardless of antibiotic presence, the evaluated processing methods, when combined with rigorous hygiene measures, maintained semen quality and reduced microbial load to the same extent as a traditional protocol using antibiotics.

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Section: Discussionmentioning

confidence: 99%

Strategies to Reduce the Use of Antibiotics in Fresh and Chilled Equine Semen

Zabala,

Serres,

Montero

et al. 2024

Animals

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“…Saturated FAs, such as C12:0 (lauric acid) and C18:0 (stearic acid), are more resistant to LPO due to the absence of carbon double bonds in their structural chains [62]. Indeed, high concentrations of lauric and stearic acids positively correlated with sperm viability and motility in stallions classified, respectively, as “good freezers” and “healthy donkeys” [63, 64]. Studies with porcine and bovine semen suggest that long-chain saturated FA can preserve sperm motility and viability by keeping ROS levels low and constituting an energy substrate for ATP production via beta-oxidation [65, 66].…”

Section: Discussionmentioning

confidence: 99%

Oxidative stress associated with proteomic and fatty acid profiles of sperm from Nellore bulls at rest

Ramírez-López,

Barros,

Vidigal

et al. 2023

Biology of Reproduction

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Sexual rest is a transient condition, which compromises conception rates, characterized by large volumes of ejaculate with high percentages of dead sperm observed in bulls. The biochemical mechanisms leading to this ejaculate pattern are not fully understood. Six adult resting Nellore bulls were submitted to Breeding Soundness Evaluation by four consecutive semen collections through the electroejaculation method during a 30 min period. Each ejaculate had its semen phenotypic parameters; morphology and physical aspects were evaluated. To assess enzymatic activity (superoxide dismutase, catalase, and glutathione S-transferase), lipid peroxidation (concentrations of malondialdehyde and nitric oxide), fatty acid, and proteomic profile aliquots of spermatozoa from the first and fourth ejaculates were used. All sperm parameters differed between the first and fourth ejaculates. Spermatozoa from the first ejaculate showed lower enzymatic activity and a higher concentration of lipid peroxidation markers. Among the 19 identified fatty acids, 52.7% are polyunsaturated. Relative abundance analysis showed that C12:0 and C18:0 fatty acids differed between the first and fourth ejaculates, being the fourth ejaculate richer in spermatozoa. The proteomics analysis identified a total of 974 proteins in both sample groups (first and fourth ejaculates). The majority of identified proteins are related to cellular processes and signaling. Quantitative proteomics showed 36 differentially abundant proteins, 6 up-regulated proteins in the first ejaculate, and 30 up-regulated proteins in the fourth ejaculate. Spermatozoa from bulls at sexual rest have less antioxidant capacity, causing changes in their fatty acid composition and protein profile, which generates the observed sperm pattern and lower fertilization capacity.

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“…Saturated FAs, such as C12:0 (lauric acid) and C18:0 (stearic acid), were more resistant to LPO due to the absence of carbon double bonds in their structural chains [40] . Indeed, high concentrations of lauric acid and stearic acid were positively correlated with sperm viability and motility in stallions classified as good freezers and healthy donkeys, respectively [41][42] . Studies with porcine and bovine semen suggest that long-chain saturated FA can preserve sperm motility and viability by keeping ROS levels low and constituting an energy substrate for ATP production via β-oxidation [43][44].…”

Section: Discussionmentioning

confidence: 98%

Association of sperm oxidative stress, proteomics and fatty acid profiles in nellore bulls at sexual rest

López¹

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Association of sperm oxidative stress, proteomics and fatty acid profiles in nellore bulls at sexual rest. Adviser: Simone Eliza Facioni Guimarães. This study was carried out with the objective of determining the effect of sexual rest on the oxidative status of spermatozoa from adult Nellore bulls and its effect on the fatty acid composition and protein profile. Six bulls were used, which were submitted to Breeding Soundness Evaluation and consecutive semen collection by the electroejaculation method. For each semen collection, the physical and morphological characteristics of the ejaculates were evaluated using conventional optical microscopy and phase contrast microscopy. After analyzing the semen qualitative parameters, the first and last ejaculate of each were centrifuged to separate the spermatozoa from the seminal plasma. From sperm aliquots, the enzymatic activity of Super Oxido Dismutase, Catalase and Glutathione S-Transferase was determined by colorimetric methods. The markers of lipid peroxidation, malondialdehyde and nitric oxide were also determined. Using gas chromatography, the fatty acids of sperm from the first and last ejaculate were identified. For proteomic analysis, total sperm proteins were extracted and precipitated in a 4% CHAPS solution, then quantified by the Bradford method and subsequently analyzed by monodimensional short-run electrophoresis and mass spectrometry (LC-MS/MS). Proteins were identified using PEAKS software version 7.0. Proteins were functionally classified by the RPSBLAST tool of BLAST version 2.13.0 into categories from the KOG database. For the label-free quantitative proteomics analysis, proteins identified in at least three technical replicates of each group with a fold-change greater than 1.5 times were included. Data were analyzed using GraphPad Prisma software version 9.1. Four semen collections were necessary so that the bulls at sexual rest presented sperm parameters compatible with reproductive fitness. Differences (p<0.05) were observed between all physical and morphological characteristics of the first and last ejaculate. A total of 974 proteins were identified, of which 735 were common between the two groups and 123 and 116 proteins were unique to spermatozoa from the first and last ejaculate, respectively. Functional classification analysis revealed that KOG categories with the highest number of classified proteins were Posttranslational modification, protein turnover, chaperones and Cytoskeleton and Signal transduction mechanisms. The protein abundance profile showed that 6 proteins were differentially abundant in spermatozoa from the first ejaculate and 30 proteins weredifferentially abundant in spermatozoa from the last ejaculate. In addition, gas chromatography analysis allowed the identification of 19 fatty acids, 47.3% of the saturated type and 52.7% of the unsaturated type. The C12:0 and C18:0 fatty acids showed difference between the two groups, being more abundant in the spermatozoa of the last ejaculate. The results indicate that oxidative stress may be related to sperm characteristics displayed by bulls at sexual rest, since the accumulation of by-products of lipid peroxidation can cause alterations in the metabolic pathways used by spermatozoa to generate energy used for motility and alter fluidity. of the membrane causing sperm abnormalities. Keywords: Bovine. Semen. Antioxidants. Lipids. Proteins.

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The Lipidomics of Spermatozoa and Red Blood Cells Membrane Profile of Martina Franca Donkey: Preliminary Evaluation

Cited by 3 publications

References 59 publications

Strategies to Reduce the Use of Antibiotics in Fresh and Chilled Equine Semen

Strategies to Reduce the Use of Antibiotics in Fresh and Chilled Equine Semen

Oxidative stress associated with proteomic and fatty acid profiles of sperm from Nellore bulls at rest

Association of sperm oxidative stress, proteomics and fatty acid profiles in nellore bulls at sexual rest

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