Abstract

Simple SummaryRam semen frequently presents with bacterial contamination, which may affect the resulting sperm vitality and fertilization ability. Since semen quality is of paramount importance for the successful artificial insemination of ewes, we focused on the description of bacterial profiles of ejaculates collected from three Slovak sheep breeds. Furthermore, we strived to unravel the effect of bacteriospermia on the immunological characteristics and oxidative profile of semen, both of which seem to play a role in bacteria-inflicted damage to male gametes.Bacterial contamination of semen is an often overlooked, yet important, factor contributing to decreased sperm vitality. Understanding the impact of bacterial presence on sperm structural integrity and functional activity may assist the development of effective strategies to prevent, or manage, bacteriospermia in the breeding practice. The aim of this study was to describe the bacterial profiles of ram semen (n = 35), and we also focused on the associations between bacteriospermia, sperm structure, and function, as well as oxidative and inflammatory characteristics of semen. For a better insight, the samples were divided into three groups, according to the breeds used in the study: native Wallachian (NW), improved Wallachian (IW), and Slovak dairy (SD) breeds. The results showed a significantly lower motility and membrane integrity in the NW group in comparison to the IW and SD groups, which was accompanied by a significantly higher concentration of leukocytes, increased reactive oxygen species (ROS) generation, and subsequent oxidative insults to the sperm lipids and proteins. Accordingly, the NW group presented with the highest bacterial load, in which Staphylococcus and Escherichia were the predominant representatives. The Pearson correlation analysis uncovered positive relationships amongst the bacterial load and leukocytospermia (r = 0.613), the extent of lipid peroxidation (r = 0.598), protein oxidation (r = 0.514), and DNA fragmentation (r = 0.638). Furthermore, positive correlations were found between the bacterial load and pro-inflammatory molecules, such as the C-reactive protein (r = 0.592), interleukin 1 (r = 0.709), and interleukin 6 (r = 0.474), indicating a possible involvement of the immune response in the process of bacteriospermia. Overall, our data indicate that ram semen quality may be equally affected by the bacterial load and diversity. Furthermore, we can assume that the presence of bacteria in ejaculates triggers inflammatory processes, causes ROS overproduction, and, thereby, contributes to alterations in the sperm structure, while at the same time compromising the fertilization ability of male gametes.

Highlights

  • Modern reproductive technologies based on artificial insemination (AI) have become an indispensable pillar of livestock production

  • Using MALDI TOF mass spectrometry, eight families, 11 genera, and 18 bacterial species were uncovered in the ram semen samples (Figure 2): Acinetobacter baylyi (A. baylyi), Aeromonas veronii (A. veronii), Bacillus subtilis (B. subtilis), Enterobacter bugandensi

  • This study reveals that the bacterial load, as well as diversity, in ram ejaculates affects the architecture and functional manifestations of male gametes, as well as the oxidative and inflammatory properties of ram ejaculates

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Summary

Introduction

Modern reproductive technologies based on artificial insemination (AI) have become an indispensable pillar of livestock production. This technique, alongside other breeding strategies, such as induction and/or synchronization of estrus and ovulation, has significantly contributed to the remarkable progress in sheep breeding programs [1]. In comparison to natural breeding, AI is more effective in adding genetic value to livestock, by spreading the use of males with valuable traits, while reducing the risks of a potential horizontal or vertical transmission of sexual diseases [2]. The success of any reproductive technology by and large depends on the initial quality of the semen sample used for the respective procedure. As reviewed by Petrovic et al [4], numerous endogenous or exogenous factors may affect the quality of ram semen. Contaminated feed and/or water, bedding, or poor hygiene standards in the breeding facility or the collection process may contribute to bacterial contamination of semen [5]

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