Comparison of different techniques to evaluate sperm membrane integrity

Bc. Adéla Grieblová

Master's thesis

Abstract:

Nowadays pig production is predominantly dependent on artificial insemination, which is almost entirely performed by liquid-stored semen. During storage, some qualitative characteristics of the spermatozoa can be reduced. An intact and functional sperm plasma membrane plays a major role in sperm fertilizing ability. Several techniques have been developed to evaluate the integrity (PMI) of the sperm plasmalemma, but there are still some controversies concerning the method that is more closely associated with sperm function. The aims of this thesis were i) to evaluate the boar sperm PMI across 72H of storage at 17 °C using eosin/nigrosin (E/N), propidium iodide/carboxyfluorescein diacetate (PI/CFDA), hypo-osmotic swelling test (HOST) and combined HOST with eosin (HOST/E), ii) to assess the correlations and the limits of agreement between these techniques, iii) and to estimate their relationships with the acrosomal status (NAR) and sperm kinetic parameters obtained by subjective and computer-assisted sperm analysis (CASA). The results showed that the PMI decreases across 72H of storage, although significant differences (p < 0.05) were found only using the HOST and HOST/E. Moreover, E/N and PI/CFDA provide significantly higher values (p < 0.05) of sperm with retained PMI than HOST and HOST/E at any incubation time. Overall, all techniques well agreed and correlated one another with the exception for the agreement between PI/CFDA and HOST. The highest correlation coefficients were found between HOST and HOST/E (r = 0.972, p < 0.001), and PI/CFDA and E/N (r = 0.708, p < 0.001). The results also showed that the integrity of the sperm head and tail membranes is associated with NAR and different kinetic traits, being the tail integrity assessed by HOST and HOST/E related to rapid linear trajectories (r = 0.332, p < 0.05 and r = 0.331, p < 0.05, respectively) and the head integrity assessed by E/N and PI/CFDA to rapid curvilinear trajectories (r = 0.325, p < 0.05 and r = 0.401, p < 0.01, respectively). The results of this thesis provide new insights into the relevance of evaluating boar sperm plasmalemma in the routine sperm analysis.

Keywords

plasma membrane, sperm lifespan, sperm viability, semen storage, swine

Jazyk práce: English

Datum vytvoření / odevzdání či podání práce: 6. 4. 2017

Obhajoba závěrečné práce

• Vedúci: Dott. Ric. Eliana Pintus
• Oponent: Eva Žatecká, Bc., externi

Czech University of Life Sciences Prague

Faculty of Agrobiology, Food and Natural Resources

Master programme / odbor:
Sustainable Agriculture and Food Security / Sustainable Agriculture and Food Security