Sperm quality and morphometry characterization of cryopreserved canine sperm in ACP-106c or TRIS

Teixeira, Diego Oliveira; Silva, Herlon Victor Rodrigues; Brito, Bruna Farias; Barbosa, Brenna de Sousa; Tabosa, Beatriz Evaristo de Almeida; Silva, Lúcia Daniel Machado da

doi:10.1590/1984-3143-AR2021-0069

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ORIGINAL ARTICLE • Anim. Reprod. 19 (3) • 2022 • https://doi.org/10.1590/1984-3143-AR2021-0069 copy

Sperm quality and morphometry characterization of cryopreserved canine sperm in ACP-106c or TRIS

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Morphological sperm evaluation supported by the morphometry can be used in the determination of the seminal quality and in the investigation of potential extenders. Although there are studies comparing TRIS and ACP extenders, there are no comparative studies between them for the computerized assisted semen analysis (CASA), sperm viability, membrane functionality and sperm morphometry parameters of cryopreserved canine semen. Hence, we aimed to evaluate the effects of ACP-106c and TRIS on post-freezing canine sperm quality. Five dogs were submitted to semen collection twice with one-week interval. The semen was evaluated within the parameters: total motility, vigor, concentration, viability, plasma membrane functionality, morphology and morphometry. In the morphometric evaluation, the morphologically normal sperm was measured as: length, width, area and perimeter of the head and the midpiece, tail length and total length. The parameters of ellipticity, elongation, regularity and roughness were determined. Then, the semen was divided into two aliquots that were diluted in TRIS or ACP-106c, with the addition of egg yolk and glycerol. The diluted semen was refrigerated and frozen. The thawed samples were evaluated. Total motility, viability, sperm membrane functionality and normal morphology reduced after thawing in both extenders (morphology reduced from 89.60 ± 1.3% to 84.40 ± 1.8 and 84.60 ± 1.1% in TRIS and ACP-106c, respectively). However, it did not differ between TRIS and ACP-106c. In the ACP-106c the sperm head defects in cryopreserved semen were higher compared to fresh semen (P < 0.05). For all the morphometric parameters evaluated, there were no differences between fresh and cryopreserved samples (3.70 ± 0.4% vs. 2.30 ± 0.5%). In kinetics, with an interval of one week statistical differences between the extenders were found only in the parameters ALH and LIN (P < 0.05). Regardless of the extender, there were no changes in the morphometric parameters of sperm after thawing.

Keywords:
cryopreservation; cryodamage; extenders; morphology; spermatozoa measurement

Parameters	Fresh	Thawed
Parameters	Fresh	TRIS	ACP-106c
Viability (%)	91.80 ± 1.6 ^a	73.30 ± 4.0 ^b	63.30 ± 4.3 ^b
Functional membrane (%)	92.80 ± 1.5 ^a	73.60 ± 3.3 ^b	63.90 ± 4.6 ^b
Normal morphology (%)	89.60 ± 1.3 ^a	84.40 ± 1.8 ^b	84.60 ± 1.1 ^b
Acrosome defect (%)	0.00 ± 0.0 ^a	0.70 ± 0.4 ^a	0.20 ± 0.1 ^a
Head defect (%)	2.30 ± 0.5 ^a	2.50 ± 0.7 ^ab	3.7 ± 0.4 ^b
Midpiece defect (%)	0.30 ± 0.2 ^a	0.80 ± 0.4 ^a	0.90 ± 0.6 ^a
Tail defect (%)	7.80 ± 1.0 ^a	11.70 ± 1.7 ^a	10.60 ± 0.9 ^a

Parameters	Fresh	Thawed
Parameters	Fresh	TRIS	ACP-106c
Head
Length	6.80 ± 0.9 ^a	6.73 ± 0.09 ^a	6.76 ± 0.9 ^a
Width	4.32 ± 0.10 ^a	4.20 ± 0.08 ^a	4.30 ± 0.11 ^a
Area	26.29 ± 0.82 ^a	25.31 ± 0.74 ^a	25.91 ± 0.93 ^a
Perimeter	19.99 ± 0.28 ^a	19.69 ± 0.28 ^a	19.90 ± 0.36 ^a
Ellipticity	1.59 ± 0.03 ^a	1.61 ± 0.03 ^a	1.59 ± 0.04 ^a
Elongation	27.67 ± 1.08 ^a	27.24 ± 1.04 ^a	27.70 ± 1.19 ^a
Rugosity	0.82 ± 0.01 ^a	0.82 ±0.01 ^a	0.82 ± 0.01 ^a
Regularity	3.54 ± 0.07 ^a	3.53 ± 0.04 ^a	3.55 ± 0.05 ^a
Midpiece
Length	10.93 ± 0.17 ^a	11.05 ± 0.14 ^a	10.89 ± 0.15 ^a
Width	1.05 ± 0.05 ^a	1.07 ± 0.06 ^a	1.05 ± 0.04 ^a
Area	13.80 ± 0.68 ^a	13.08 ± 0.63 ^a	13.01 ± 0.69 ^a
Perimeter	24.86 ± 0.16 ^a	24.69 ± 0.24 ^a	24.56 ± 0.20 ^a
Tail length	45.49 ± 0.43 ^a	45.03 ± 0.31 ^a	44.42 ± 0.40 ^a
Total length of sptz	63.23 ± 0.50 ^a	62.81 ± 0.42 ^a	62.08 ± 0.48 ^a

Parameters	TRIS	ACP-106c
Total motility (%)	47.95 ± 7.0 ^a	42.09 ± 7.0 ^a
Progressive motility (%)	10.81 ± 2.2 ^a	8.70 ± 2.6 ^a
Average path velocity (µm/s)	67.15 ± 3.5 ^a	68.38 ± 3.7 ^a
Straight line velocity (µm/s)	60.90 ± 3.6 ^a	63.24 ± 3.9 ^a
Curvilinear velocity (µm/s)	88.81 ± 5.1^a	86.61 ± 4.5 ^a
Amplitude of lateral head displacement (µm)	3.66 ± 0.2 ^a	2.91 ± 0.2 ^b
Beat cross frequency (Hz)	11. 53 ± 0.5 ^a	11.23 ± 0.6 ^a
Straightness (%)	90.42 ± 1.0 ^a	92.16 ± 1.2 ^a
Linearity (%)	68.68 ± 1.0 ^b	73.11 ± 2.6 ^a
Wobble coefficient (%)	75.97 ± 1.1^a	79.15 ± 2.1 ^a

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[1] *Corresponding author: lucia.daniel@uece.br