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Research article - Peer-reviewed, 2023

Single layer centrifugation (SLC) for bacterial removal with Porcicoll positively modifies chromatin structure in boar spermatozoa

Lacalle, Estibaliz; Fernandez-Alegre, Estela; Soriano-ubeda, Cristina; Martinez-Martinez, Sonia; Dominguez, Juan Carlos; Gonzalez-Montana, J. Ramiro; Morrell, Jane M.; Martinez-Pastor, Felipe

Abstract

The storage of boar semen samples at 17 degrees C for artificial insemination (AI) doses enables the proliferation of the bacteria, making antibiotics necessary. This can contribute to the development of antimicrobial resistance (AMR). This study tested bacterial presence and sperm chromatin structure after using a low-density colloid (Porcicoll) as an antibiotic alternative to eliminate bacteria. Ejaculates (8 boars, 3 ejac-ulates each) were split as control and low-density colloid centrifugation (single layer centrifugation, SLC, 20%, and 30% Porcicoll) into 500 ml tubes. Analyses were carried out at days 0, 3, and 7 (17 degrees C) for microbial presence and sperm chromatin structure analysis: %DFI (DNA fragmentation) and %HDS (chromatin immaturity), monobromobimane (mBBr; free thiols and disulfide bridges), and chromomycin A3 (CMA3; chromatin compaction). Besides comparing bacterial presence (7 species identified) and chromatin variables between treatments, the associations between these sets of variables were described by canonical correlation analysis (CCA). Results showed a significant decrease of some bacteria or a complete removal after SLC (especially for P30). SLC also caused a decrease of %HDS and an increase of disulfide bridges and low and medium mBBr populations, suggesting the removal of immature sperm (poor chromatin compaction). CCA showed an association pattern compatible with the degradation of sperm chromatin parameters with bacterial contamination, especially Enterobacteria, P. aeuriginosa, and K. variicola. In conclusion, bacterial contamination affects sperm chromatin beyond DNA fragmentation; SLC with low-density colloid not only removes bacteria from boar semen, but also chromatin structure is enhanced after selection.(c) 2023 Published by Elsevier Inc.

Keywords

Boar; DNA fragmentation; Chromatin; Canonical correlation analysis; Single layer centrifugation

Published in

Theriogenology
2023, Volume: 201, pages: 95-105
Publisher: ELSEVIER SCIENCE INC

    Associated SLU-program

    AMR: Bacteria

    Sustainable Development Goals

    SDG3 Good health and well-being

    UKÄ Subject classification

    Clinical Science

    Publication identifier

    DOI: https://doi.org/10.1016/j.theriogenology.2023.02.017

    Permanent link to this page (URI)

    https://res.slu.se/id/publ/121824