The quality of great scallop (Pecten maximus) sperm after thawing

Abstract

Most publications devoted to the cryopreservation of mollusc sperm have focused on the definition of technical protocols, avoiding the description of sperm quality after thawing. The present study investigated the effects of cryopreservation on sperm quality in the great scallop. Wild scallop were fished during the natural spawning period and conditioned in the hatchery before use. Sperm samples were obtained after intragonadal injection of serotonin and cryopreserved using a previously published protocol. Sperm quality was assessed using a panel of four parameters: sperm motility characteristics, using a computer assisted sperm analysis plugin with Image J, intracellular ATP content using an ATP-Lite kit, sperm integrity, using flow cytometry and sperm morphology, using transmission electron microscopy. For each parameter, fresh (control) and thawed spermatozoa were compared. A significant decrease of both the percentage of motile spermatozoa (reduction: 75%) and sperm swimming speed (86%) were observed for thawed sperm compared with fresh sperm. The percentage of living spermatozoa, as assessed using flow cytometry, was significantly lower for thawed sperm (72.4±2.5%) compared with fresh sperm (86.4±1.1). However, no significant difference of intracellular sperm ATP content was observed between fresh and thawed sperm. Post thawing, while some spermatozoa showed little or no morphological differences compared with fresh sperm, others had undergone drastic changes, including swelling of the plasma membrane, structural alterations of the chromatin and damage to mitochondria. In conclusion, the descriptive parameters studied in the present work showed that the quality of thawed great scallop sperm was lower than that of fresh cells but was still sufficient for use in aquaculture programs and sperm cryobanking for this species.

Quality assessment of cryopreserved black-lip pearl oyster Pinctada margaritifera spermatozoa

“…We observed swelling in the plasma membrane after the freeze/thaw process. Similar damage was reported in the Chilean scallop Argopecten purpuratus (Espinoza et al 2010), blue mussel Mytilus galloprovincialis (Liu et al 2016) and great scallop Pecten maximus (Suquet et al 2016).…”

“…In the active spermatozoa, we observed 20% lower VAP in frozen/thawed sperm compared to fresh. Suquet et al (2016) reported 75% lower velocity in frozen/thawed spermatozoa of the great scallop compared to fresh. Linhart et al (2005) (Figueroa et al 2016), 97 µm s -1 to 134 µm s -1 in Prochilodus lineatus (Viveiros and Leal 2016;Viveiros et al 2017), 57 µm s -1 in rainbow trout Oncorhynchus mykiss (Suquet et al 2012b) and µm s -1 in Siberian sturgeon Acipenser baerii spermatozoa (Sarosiek et al 2014).…”

“…Fertilization of an oocyte is dependent on the quality of spermatozoa, specifically their motility (Cosson et al 2008a, b) and metabolic activity (mitochondrial respiration, ATP hydrolysis) (Boulais et al 2015(Boulais et al , 2017Suquet et al 2016). It has been reported that cryopreservation may induce oxidative stress (Li et al 2010a;Shaliutina-Kolešová et al 2015) damaging spermatozoon structure (Gwo et al 2003;Paniagua-Chavez et al 2006;Suquet et al 2016) and decreasing motility (Dong et al 2005;Gwo et al 2002;Riesco et al 2017;Suquet et al 2016) and fertilizing capacity (Boulais et al 2017;Dong et al 2005;Gwo et al 2002;Narita et al 2008;Paniagua-Chavez and Tiersch. 2001;Riesco et al 2017).…”

“…Mytilus galloprovincialis (Di Matteo et al 2009;Liu et al 2016), Argopecten purpuratus (Espinoza et al 2010), Ostrea edulis (Horváth et al 2012), Pecten maximus (Suquet et al 2016), and Pinctada angulata (Riesco et al 2017). Lyons et al (2005) and Acosta-Salmon et al (2007) reported the effect of several cryoprotectant additives on P. margaritifera sperm motility after freeze/thawing and Hui et al (2011) reported reduced viability of spermatozoa after freeze/thawing.…”

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

“…These studies concerned oyster species (Boulais et al, ; Suquet et al, ; Suquet, Rimond, et al, ), pearl oyster species (Demoy‐Schneider, Levêque, Schmitt, Le Pennec, & Cosson, ; Demoy‐Schneider, Schmitt, Le Pennec, Suquet, & Cosson, ; Zheng et al, ) and so on. However, studies for scallop species were limited to Pecten maximus (Suquet, Arsenault‐Pernet, Ratiskol, & Mingant, ; Suquet, Gourtay, et al, ; Suquet, Malo, et al, ; Suquet, Quéré, et al, ), Argopecten purpuratus (Lamarck, 1819) (Dupré, Covarrubias, Goldstein, Guerrero, & Rojas, ; Espinoza, Valdivia, & Dupre, ), Chlamys farreri (Li, Li & Xue, 2000; Xue, ) and Patinopecten yessoensis (Yang et al, ; Yang, Yang, Liu, & Zhou, , ). Because of the difference in species, spermatozoa cryopreservation methods and related parameters in different species are different.…”