Sucrose increases the quality and fertilizing ability of cryopreserved chicken sperms in contrast to raffinose

Abstract

Chicken semen conservation is an important tool for programs of genetic diversity management and of endangered breeds’ conservation. However, the method still needs to be improved in order to be applied in a wide variety of environments and breeds. Our objective was to compare the effects of 2 external cryoprotectants saccharides (sucrose and raffinose) on the sperm freezability of a Thai local breed, Pradu Hang Dum, in which semen was frozen with a simple freezing method using nitrogen vapors and dimethyl formamide (DMF). Thirty-six males were selected on their motility vigor score for the experiments. In a first experiment, a large range of sucrose and raffinose doses were tested. Semen quality was evaluated after incubation at 5°C or after cryopreservation in straws in the saline Blumberger Hahnen Sperma Verdünner diluent + DMF (6% v/v) with or without sucrose/raffinose. The best targeted doses of sucrose and raffinose were then kept for experiment 2 that was focused on cryopreserved semen. In this experiment, semen quality was measured on frozen-thawed sperm: different objective motility data evaluated by computer-assisted sperm analysis (CASA), membrane integrity, acrosome integrity, mitochondria function evaluated using flow cytometry, lipid peroxide production assessed by the thiobarbituric acid test. Fertility obtained with frozen-thawed semen supplemented or not with sucrose or raffinose was also evaluated after artificial insemination of laying hens. The presence of sucrose at the osmotically inactive dose 1 mmol significantly increased the vigor motility, membrane integrity, acrosome integrity, and mitochondrial functions of frozen-thawed sperm ( P < 0.05), and showed the highest levels of fertility after sperm cryopreservation (91% vs. control 86%, P < 0.001). Raffinose showed negative effects on in vitro semen quality from 1 to 100 mmol. Fertility was also negatively ( P < 0.001) affected by raffinose (fertility rate 66 to 70%). We thus showed in the present study the high success of a simple chicken sperm cryopreservation method with an external cryoprotectant easily available and cheap, the sucrose, used at an osmotically inactive low concentration.

Validation of the Turkey Semen Cryopreservation by Evaluating the Effect of Two Diluents and the Inseminating Doses

“…In addition, Lake includes the polyvinylpyrrolidone (PVP), whereas this is absent in Tselutin. The PVP is a water-soluble polymer, which is identified as a NP-CPA at a high molecular weight by various authors [4,13,[30][31][32]. Thus, we can speculate that there is a synergistic effect between PVP and Ficoll that offers more effective dehydration to the sperm cells during the freezing process.…”

Effects of Saccharides Supplementation in the Extender of Cryopreserved Rooster (Gallus domesticus) Semen on the Fertility of Frozen/Thawed Spermatozoa

“…One way to solve the problem of low fertility is to improve the cryoprotective environment for rooster semen with components that will prevent spermatozoon death and damage during freezing, storage, and thawing of the semen [ 9 , 10 ]. According to Thananurak (2019), a balanced cryopreservation diluent is the key to maintaining the functionality of sperm after thawing [ 11 ].…”

“…To preserve its structure when exposed to negative temperatures, the protein must be embedded in a rigid glassy matrix [ 13 ]. Saccharose [ 11 ] and trehalose [ 14 ] have been used as such components, performing a cryoprotective function in mammalian semen media. Disaccharides, especially trehalose and sucrose, are widely known as effective bioprotective agents with exceptional ability to neutralize cold stress [ 15 , 16 ].…”

Comparative transcriptome analysis digs out genes related to antifreeze between fresh and frozen–thawed rooster sperm

“…But sperm freezing for poultry is very limited because of its lower fertility and high individual within-breed variability ( Blesbois, 2007 , Chuaychu-Noo et al., 2016 ). However, sperm cryopreservation in poultry has a great potential of application in many fields, including artificial insemination practice, breeding selection, disease prevention, the conservation of genetic resources, and ex situ management of avian genetic resources ( Thananurak et al., 2019 , Thélie et al., 2019 ), which are of great economic importance for the commercial poultry industry. The cryopreservation of avian semen began in 1942 ( Shaffner, 1942 ) but failed to achieve hatched chicks until 1949 ( Polge et al., 1949 ).…”