Bacterial DNase activity as a putative inductor of sperm DNA fragmentation in infected bull frozen-thawed semen samples

被引:1
|
作者
Gosalvez, Jaime [1 ]
Bartolome-Nebreda, Javier [1 ,2 ]
Tvrda, Eva [3 ]
Lopez-Fernandez, Carmen [1 ]
Luis Fernandez, Jose [4 ,5 ]
Johnston, Stephen [6 ]
机构
[1] Univ Autonoma Madrid, Dept Biol, Unit Genet, Campus Cantoblanco, Madrid 28049, Spain
[2] Halotech DNA, PCM Parque Cient Madrid, Madrid 28049, Spain
[3] Slovak Univ Agr, Fac Biotechnol & Food Sci, Inst Appl Biol, Tr A Hlinku 2, Nitra 94976, Slovakia
[4] Ctr Oncol Galicia, Mol Genet & Radiobiol Lab, La Coruna 15009, Spain
[5] INIBIC Complejo Hosp Univ A Coruna CHUAC, Genet Unit, La Coruna, Spain
[6] Univ Queensland, Sch Agr & Food Sci, Gatton, Australia
关键词
Bovine frozen -thawed semen; Sperm DNA fragmentation; DNase activity; Bacterial contamination; Sperm chromatin dispersion assay; EXTRACELLULAR DEOXYRIBONUCLEASE; IDENTIFICATION; CONTAMINATION;
D O I
10.1016/j.theriogenology.2022.12.011
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The aim of this study was to investigate the relationship between DNase activity associated with bacterial contamination of incubated bovine frozen-thawed spermatozoa and elevated sperm DNA fragmentation. Electrophoresis analysis of plasmid PBR322 incubated for 30 min at 37 degrees C with the supernatant of the diluent of frozen-thawed centrifuged bovine semen straws infected with bacteria showed clear evidence of DNase activity when compared to plasmid incubated in similarly prepared non-infected bovine diluent supernatant (Experiment 1). This DNase activity was subsequently found to be time dependent (0-60 min) and its activity prevented in the presence of EDTA (10 and 20 mM; Experiment 2). Semen straws infected (n = 10) and not infected (n = 10) with bacteria where incubated at 37 degrees C for up to 48h post-thaw. Semen infected with bacteria showed an exponential increase in bacterial growth and a corresponding increase in sperm DNA fragmentation. Non-infected semen samples showed no change in the incidence of sperm DNA fragmentation over the same period of incubation (Experiment 3). Our experiments reinforce the idea that exogenous DNases present in the semen should be considered as one of the primary contributing causes of sperm DNA fragmentation post ejaculation. In the case of the bull, post-thaw incubation of commercial straws contaminated with bacteria, resulted in increased levels of sperm DNA fragmentation, most likely associated with DNase activity (potentially restriction endonu-cleases) derived from the bacteria. Such adverse changes in sperm DNA fragmentation, as described here in vitro, may be also operative after insemination in the female reproductive tract (in vivo) and highlight the importance of implementing high levels of hygiene practice during semen processing, especially in light of future trends of bacterial resistance to the common antibiotics used in semen diluents.(c) 2022 Elsevier Inc. All rights reserved.
引用
收藏
页码:224 / 231
页数:8
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