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

Abstract

This study was designed to test the fertilizing ability of cryopreserved turkey semen, and here, two experiments were performed: an in vitro analysis to assess the effects of Tselutin and Lake diluents and an in vivo test to determine the fertility and hatching rates by also studying the feat of three insemination doses (250, 400 and 600 × 106 sperm/hen). Pooled semen samples were diluted with Tselutin or Lake extender which contained 20% of dimethylsulfoxide and 1 mM of Ficoll at final sperm concentration of 3 × 109 sperm/mL. Thereafter, semen was packaged into straws and frozen on liquid nitrogen. The post-thaw sperm quality was evaluated considering motility (computer-aided sperm analysis—CASA system) and membrane integrity (flow cytometry). Significantly higher values of progressive motility and some kinetic parameters in semen frozen with Lake were found. When we compared the extenders in vivo, no significant effects were detected, whilst sperm concentration significantly affected both fertility and hatching rates, with the best results obtained with the sperm concentration of 400 × 106 sperm/hen. From the results obtained, it emerged that the extender type only affected sperm motility characteristics, not the fertilizing ability of frozen-thawed semen, while inseminating dose markedly affected fertility and hatching rates.

Effect of freeze‐dried quail egg white and yolk addition to semen extender on viability of rooster sperm stored for 6 h at 4°C

“…It is believed that the usefulness of semen for artificial insemination purposes can be assessed by sperm motility, which should be higher than 80% (Di Iorio et al, 2020). Numerous scientific reports…”

“…Semen dilution is a very important initial step of in vitro processing, storage and preservation for efficient artificial insemination (Smith et al, 2018). Numerous studies indicate a significant influence of diluents and storage time on sperm morphology and motility parameters in chicken (Donoghue & Wishart, 2000; Dumpala et al, 2006; Hudson et al, 2016; Iaffaldano et al, 2008; Łukaszewicz, Jerysz, & Kowalczyk, 2020; Mphaphathi et al, 2016; Santiago‐Moreno et al, 2012; Siudzińska & Łukaszewicz, 2008; Vasicek et al, 2015), turkey (Di Iorio et al, 2020; Słowińska et al, 2018), guinea fowl (Hudson et al, 2016; Keerthy et al, 2016), ostrich (Smith et al, 2018), capercaillie (Kowalczyk et al, 2012), quail (Chełmońska et al, 2006), duck (Łukaszewicz, Jerysz, & Chełmońska, 2020) and goose (Łukaszewicz, 2002a, 2002b). Semen preservation is also a very important tool necessary in rescue and conservation of bird species and breeds.…”

From the Semen Collection Method to the Hatchlings: The Use of Cryopreserved Sperm from Pheasants Fed an Antioxidant-Enriched Diet

“…Comparable fertility values (26–37%) for F–T sperm have been reported under similar AIs conditions [ 27 , 45 ]. In the Combatiente chicken, doses of 500 × 10 6 F–T sperm had a reported fertility of just 9.4% [ 72 ], whereas 87% fertility has been achieved in the turkey with doses of 400 × 10 6 F–T sperm [ 73 ]. Increasing the sperm dose in the above-described AI protocol for pheasants would probably result in higher fertility percentages.…”

Comparison of Commercial Poultry Semen Extenders Modified for Cryopreservation Procedure in the Genetic Resource Program of Czech Golden Spotted Hen

“…(SAS Institute Inc., Cary, NC, USA). The general linear model (GLM) procedure was used to analyze variables after a check of residual distribution (residual QQ plot) [ 14 , 25 ]. A statistical model with two fixed effects of the day of semen collection (5 classes with 18 observations per class) and modified extenders (3 classes with 30 observations per class) was applied to cover the variability among dependent variables.…”

“…For this reason, supplementation with cryoprotectants that preserve the cells during the freezing process represents an important strategy in the modification process [ 4 , 5 ]. Some of the most used cryoprotectants include glycerol [ 6 , 7 ], N-methylacetamide (NMA) [ 8 ], dimethylacetamide [ 9 , 10 , 11 , 12 ] or dimethyl sulfoxide [ 13 , 14 ]. The glycerol addition to IDs reduces the fertilizing ability of sperm.…”