Bussalleu Eva, Sancho Sílvia, Briz Maria D, Yeste Marc, Bonet Sergi
Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Catalonia, Spain.
Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Catalonia, Spain.
Theriogenology. 2017 Feb;89:235-243. doi: 10.1016/j.theriogenology.2016.11.017. Epub 2016 Nov 18.
The use of antimicrobial peptides (AMP) has become one of the most promising alternatives to the use of antibiotics (Abs) in semen extender's formulation to overcome the increasing bacterial resistance to antibiotics. However, AMP may impair boar sperm quality, so that their deleterious effects might be higher than their effectiveness against bacteria. Thus, the aim of this study was to determine whether three different AMP, the proline-arginine-rich antimicrobial peptide PR-39 (PR-39), and the porcine myeloid antimicrobial peptides 36 (PMAP-36) and 37 (PMAP-37) had any effect upon boar sperm quality and bacterial growth. For this purpose, three different concentrations of each peptide (1 μM, 10 μM and 20 μM for PR-39 and 0.5 μM, 1 μM and 3 μM for PMAP-36 and PMAP-37) were added to 2 mL of a pool of extended semen with BTS without Abs; two controls, one without AMPs and Abs, and the other with Abs only were used for each peptide (n = 3). Total (TMOT) and progressive (PMOT) sperm motility, sperm viability and bacterial concentration were assessed before the addition of each AMP or Abs and at 1, 3, 6, 8 and 10 days post-addition. For each AMP, results revealed a drop in the TMOT and PMOT in all treatments and controls. In regard to sperm viability, while PR-39 at 10 μM maintained it in values similar to those of the control with Abs and PMAP-36 kept also the sperm viability in a similar fashion to the treatment with Abs, PMAP-37 was more effective in keeping sperm viability than controls (P < 0.05). Whereas PR-39 at 20 μM and PMAP-37 at 3 μM were quite effective in controlling bacterial load, PMAP-36 did not avoid bacterial growth at any concentration tested. In conclusion, taking all results together, PMAP-37 seems to be a suitable candidate to replace Abs in extended semen, as it hardly impairs sperm viability and controls the bacterial load. Nevertheless, further studies are still required to improve its effectiveness.
在精液稀释剂配方中使用抗菌肽（AMP）已成为最有前景的替代抗生素（Abs）的方法之一，以克服细菌对抗生素日益增加的耐药性。然而，AMP可能会损害公猪精子质量，因此其有害影响可能高于其对细菌的有效性。因此，本研究的目的是确定三种不同的AMP，即富含脯氨酸-精氨酸的抗菌肽PR-39、猪髓样抗菌肽36（PMAP-36）和37（PMAP-37）对公猪精子质量和细菌生长是否有任何影响。为此，将每种肽的三种不同浓度（PR-39为1μM、10μM和20μM，PMAP-36和PMAP-37为0.5μM、1μM和3μM）添加到2mL不含抗生素的BTS稀释精液池中；每种肽使用两个对照，一个不含AMP和抗生素，另一个仅含抗生素（n = 3）。在添加每种AMP或抗生素之前以及添加后1、3、6、8和10天评估总精子活力（TMOT）、前向精子活力（PMOT）、精子存活率和细菌浓度。对于每种AMP，结果显示所有处理和对照中的TMOT和PMOT均下降。关于精子存活率，虽然10μM的PR-39将其维持在与含抗生素对照相似的值，并且PMAP-36也以与含抗生素处理相似的方式保持精子存活率，但PMAP-37在保持精子存活率方面比对照更有效（P < 0.05）。虽然20μM的PR-39和3μM的PMAP-37在控制细菌载量方面相当有效，但PMAP-36在任何测试浓度下都不能避免细菌生长。总之，综合所有结果，PMAP-37似乎是在稀释精液中替代抗生素的合适候选物，因为它几乎不损害精子存活率并控制细菌载量。然而，仍需要进一步研究以提高其有效性。
