Abstract
Simple SummaryOverbreeding—and its associated increase in the chest areas of turkeys—has led to the loss of their natural ability to reproduce. Therefore, commercial production of turkey meat relies on artificial insemination. However, along with the physiology of the genital tract of turkeys, there is high potential for bacterial contamination of ejaculates. These bacteria may affect crucial semen quality parameters required for successful fertilization. As such, it is important to pay close attention to the bacteria present in turkey ejaculates and possible solutions to eliminate their adverse effects on avian spermatozoa.This study focused on the identification of naturally occurring bacteria in the reproductive fluid and impact on the quality of ejaculates obtained from the turkey breed British United Turkeys (BUT) Big 6 (n = 60). We determined possible relationships between the bacterial load and advanced sperm quality parameters that are important for effective artificial insemination and high fertility, as well as the concentration of selected antimicrobial proteins and pro-inflammatory markers of turkey semen. Sperm motility was assessed with computer-assisted sperm analysis (CASA), while the membrane and acrosome integrity were examined with smearing and staining methods. Reactive oxygen species (ROS) generation was quantified via luminometry, sperm DNA fragmentation was evaluated using the TUNEL assay, and the JC-1 assay was applied to evaluate the mitochondrial membrane potential. Cell lysates were prepared to investigate the extent of lipid and protein oxidation. Furthermore, levels of interleukins 1 and 6 (IL-1, IL-6), C-reactive protein, cathelicidin, and β-defensin were quantified in the seminal plasma using the ELISA method. The most dominant species identified by the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was Escherichia coli, Proteus mirabilis, Staphylococcus lentus, and Citrobacter braakii. The bacterial load had a negative effect on the sperm motility (p < 0.001), as well as membrane (p < 0.05) and acrosome integrity (p < 0.01). A strong positive relationship between the bacterial load and DNA fragmentation (p < 0.001) was detected as well. Positive associations were recorded between the increasing presence of bacteria, ROS overgeneration (p < 0.001), and a subsequent oxidative damage to the proteins (p < 0.001) and lipids (p < 0.01). It was revealed that the antimicrobial peptides β-defensin (p < 0.001) and cathelicidin (p < 0.001) had a positive relationship with the motility. In contrast, pro-inflammatory markers, such as IL-1 (p < 0.001) and IL-6 (p < 0.001), had a negative impact on the motion behavior of turkey spermatozoa. Our results suggest that the semen quality may be notably affected by the bacterial quantity as well as quality. It seems that bacteriospermia is associated with inflammatory processes, oxidative stress, sperm structural deterioration, and a subsequent risk for a failed artificial insemination in turkey breeding.
Highlights
The global poultry industry has made enormous progress over the last three decades, in the field of meat and egg production [1]
Using MALDI-TOF mass spectrometry, 7 families, 12 genera, and 17 bacterial species were identified in turkey ejaculates (Figure 1): Bacillus cereus (B. cereus), Bacillus subtilis (B. subtilis), Citrobacter braakii (C. braakii), Empedobacter brevis (E. brevis), Enterococcus faecium (E. faecium), Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae), Morganella morganii (M. morganii), Myroides odoratimimus (M. odoratimimus), Proteus hauseri (P. hauseri), Proteus mirabilis (P. mirabilis), Proteus penneri (P. penneri), Proteus vulgaris (P. vulgaris), Staphylococcus chromogenes (S. chromogenes), Staphylococcus lentus (S. lentus)
As the industrial production of turkeys relies on artificial insemination, it is extremely important to pay attention to the quality of semen intended for assisted reproduction
Summary
The global poultry industry has made enormous progress over the last three decades, in the field of meat and egg production [1]. In favor of, for example, muscle incrementation in the chest area, has caused significant differences between males and female turkeys This disparity has led to the loss of the turkey’s natural ability to reproduce; this is due to the male’s inability to physically approach the female’s cloaca in order for fertilization to transpire. A key component of the innate immune system of birds encompasses a group of antimicrobial peptides, such as β-defensins and cathelicidins. In addition to their antibacterial properties, these peptides may regulate the production of cytokines and, selectively support the host’s immune responses [9]
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