Morphological alterations in cryopreserved spermatozoa of scallop Argopecten purpuratus

Abstract

ABSTRACT. The present work identifies and quantifies the morphological alterations of scallop Argopecten purpuratus spermatozoa caused by long-term cryopreservation. Percentages of motility, fertilization and injured spermatozoa were quantified by optic microscopy and scanned electron microscopy. These parameters were evaluated in sperm without treatment (CTR), spermatozoa incubated in cryoprotective solution but not freezed (ICS) and freezed-thawed spermatozoa (FTS). Spermatozoa of ICS treatment remained motile longer than those of CTR, whereas those of FTS treatment were lowest. Morphology of the spermatozoa was affected in several ways by the freeze-thawing treatment; some had their head deformed or swollen, others had their cell membrane folded or broken; acrosome reaction; anomalous positions or absence of mitochondria as well as broken, stiff or loss of lineal structure of tail. CTR and ICS treatments had higher percentages of undamaged sperm (87.7% and 79.0% respectively), while FTS samples had 14.2% of undamaged sperm. The tail was the spermatic structure most commonly injured in FTS (77.0%), the percentage of sperm with head injury was 55.1% and with acrosome reaction was 28.7%, whereas middle piece was affected in 23.9% of sperm. Percentages of fertilization were 68.3%, 67.9% and 58.2% for CTR, ICS and FTS respectively, which were not significantly different. There was a higher correlation between injuries and motility than between injuries and fertilization success. Correlation between motility and fertilization was low (0.605 and 0.668 with motility at 5 and 30 min, respectively). Keywords: cryopreservation, Argopecten purpuratus, scallop, sperm, Chile. Alteraciones morfológicas en espermatozoides criopreservados de concha de abanicoArgopecten purpuratus RESUMEN. El presente trabajo identifica y cuantifica las alteraciones morfológicas en espermatozoides de concha de abanico A. purpuratus causadas por la criopreservación en nitrógeno líquido. Porcentajes de motilidad, fecundación de ovocitos frescos y espermatozoides lesionados (en cabeza, acrosoma, pieza media y flagelo) fueron determinados bajo microscopía óptica y electrónica de barrido. Estos parámetros fueron evaluados en un control sin tratamiento (CTR), espermatozoides incubados en solución crioprotectora pero sin congelamiento (ICS) y espermatozoides congelados-descongelados (FTS). Los espermatozoides del tratamiento ICS mostraron mayor motilidad que de CTR, mientras que la motilidad de los espermatozoides del tratamiento FTS fue la más baja. La morfología externa de los espermatozoides fue afectada de varias formas por el congelamiento-descongelamiento; cabeza deformada o hinchada, membrana celular plegada o rota, reacción acrosómica, anormal posición o ausencia de las mitocondrias, ruptura, rigidez o pérdida de la estructura lineal del flagelo. El CTR e ICS presentó los mayores porcentajes de espermatozoides ilesos (87,7% y 79,0% respectivamente), mientras que las muestras de FTS tuvieron 14,2% de espermatozoides ilesos. El flagelo fue la e...

Cryopreservation of strip spawned sperm using non-programmable freezing technique in the blue musselMytilus galloprovincialis

“…; Hui, Vonau, Moriceau, Tetumu, Vanaa, Demoy‐Schneider, Suquet & Moullac ), greenshell mussels (Smith et al . ), scallops Argopecten purpuratus (Espinoza, Valdivia & Dupré ), farmed greenlip and black‐lip abalone (Liu, Li et al . ; Liu, Xu et al .…”

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.…”

“…These studies concerned oyster species (Boulais et al, 2018;Suquet et al, 2012;Suquet, Rimond, et al, 2013), pearl oyster species (Demoy-Schneider, Levêque, Schmitt, Le Pennec, & Cosson, 2012;Zheng et al, 2018) and so on. However, studies for scallop species were limited to Pecten maximus (Suquet, Arsenault-Pernet, Ratiskol, & Mingant, 2014;Suquet, Quéré, et al, 2013), Argopecten purpuratus (Lamarck, 1819) (Dupré, Covarrubias, Goldstein, Guerrero, & Rojas, 2016;Espinoza, Valdivia, & Dupre, 2010), Chlamys farreri (Li, Li & Xue, 2000;Xue, 1994) and Patinopecten yessoensis (Yang et al, 1999;Yang, Yang, Liu, & Zhou, 2007.…”

Improved solution for vitrification ofProchilodus lineatusembryos based on the reduction in risk factors: Toxicity, osmotic responses and ice-nucleation

“…In addition to ice crystal formation, low temperatures generate changes in cellular metabolism (Dinnyés, Urbanyi, Baranyai & Magyary, ; Espinoza, Valdivia & Dupré, ; Zhang & Rawson, ), exerting a great influence on ontogenetic development, degrading genetic material, and denaturing important proteins for embryonic development (Gwo et al., ; Zilli, Schiavone, Zonno, Storelli & Vilella, ; Zilli & Vilella, ). The sensitivity to cold is characteristic to each species and cell type, and is commonly higher at early stages of development, becoming lower during developmental processes (Ninhaus‐Silveira et al., ; Robles, Cabrita, Real, Álvarez & Herráez, ; Zhang & Rawson, ).…”