Post-thaw quality of ram sperm frozen with different concentrations of low-density lipoproteins associated with non-enzymatic antioxidants

Snoeck, Paola Pereira das Neves; Câmara, Diogo Ribeiro; Moura, Luís Cláudio de Oliveira; Silva, Maíra Corona da; Machado-Neves, Mariana; Teixeira-Neto, Milton Rezende; Henry, Marc

doi:10.1590/1984-3143-AR2022-0068

Paola Pereira das Neves Snoeck ^**Corresponding author: paolasnoeck@uesc.br

conceptualization

methodology

supervision

writing review and editing

Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil

https://orcid.org/0000-0003-4445-8630

Diogo Ribeiro Câmara

writing original draft

writing review and editing

Departamento de Medicina Veterinária, Universidade Federal de Alagoas, Viçosa, AL, Brasil

https://orcid.org/0000-0002-9873-7419

Luís Cláudio de Oliveira Moura

data curation

formal analysis

methodology

Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil

https://orcid.org/0000-0002-6995-9864

Maíra Corona da Silva

formal analysis

methodology

Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil

https://orcid.org/0000-0003-3455-2789

Mariana Machado-Neves

conceptualization

writing review editing

Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, MG, Brasil

https://orcid.org/0000-0002-7416-3529

Milton Rezende Teixeira-Neto

data curation

formal analysis

Departamento de Medicina Veterinária, Centro Universitário UNIFTC, Vitória da Conquista, BA, Brasil

https://orcid.org/0000-0002-6516-6322

Marc Henry

conceptualization

funding acquisition

supervision

Departamento de Clínica e Cirurgia Veterinárias, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil

# (In Memoriam)

https://orcid.org/0000-0003-4730-8128

#
(In Memoriam)

*Corresponding author: paolasnoeck@uesc.br

Conflicts of interest: The authors have no conflict of interest to declare.

Author contributions

PPNS: Conceptualization, Methodology, Supervision, Writing-review & editing; DRC: Writing- original draft, Writing-review & editing; LCOM: Data Curation, Formal Analysis, Methodology; MCS: Formal Analysis, Methodology; MMN: Conceptualization, Writing-review & editing; MRTN: Data curation, Formal Analysis; MH: Conceptualization, Funding acquisition, Supervision.

Figures (2)
Tables (4)

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Figure 1
Experiment I ram sperm frozen in Tris-glucose base extender containing 16% egg yolk and 5% glycerol (Control, C) or using base extender with no egg yolk, 5% glycerol and 2% low-density lipoprotein (G5L2) with addition of different non-enzymatic antioxidants (10 mM). Structural (IP^-CFDA⁺) and functional integrity (hypoosmotic test) post-thaw. AA: ascorbic acid; HB: hydroxytoluene butylate; AP: ascorbyl palmitate; TH: trehalose. Different letters indicate differences among treatments (P < 0.05).

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Figure 2
Experiment II ram sperm frozen in Tris-glucose base extender containing 16% egg yolk and 5% glycerol (Control, C) or using base extender with no egg yolk, 5% glycerol and 8% low-density lipoprotein (G5L8) with addition of different non-enzymatic antioxidants (10 mM). Structural (IP^-CFDA⁺) and functional integrity (hypoosmotic test) post-thaw. AA: ascorbic acid; HB: hydroxytoluene butylate; AP: ascorbyl palmitate; TH: trehalose. Different letters indicate statistical differences between treatments (P < 0.05).

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Table 1
Post-thaw kinematic parameters of ram sperm (mean ± SEM) frozen in Tris-glucose base extender containing 16% egg yolk and 5% glycerol (Control) or using base extender with no egg yolk, 5% glycerol and 2% low-density lipoprotein (G5L2) with addition of different non-enzymatic antioxidants (10 mM).

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Table 2
Kinematic parameters of ram sperm (mean ± SEM) frozen in Tris-glucose base extender containing 16% egg yolk and 5% glycerol (Control) or using base extender with no egg yolk, 5% glycerol and 2% low-density lipoprotein (G5L2) with addition of different non-enzymatic antioxidants (10 mM) during 3h of incubation at 38 °C after thawing.

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Table 3
Post-thaw kinematic parameters of ram sperm (mean ± SEM) frozen in Tris-glucose base extender containing 16% egg yolk and 5% glycerol (Control) or using base extender with no egg yolk, 5% glycerol and 8% low-density lipoprotein (G5L8) with addition of different non-enzymatic antioxidants (10 mM).

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Table 4
Kinematic parameters of ram sperm (mean ± SEM) frozen in Tris-glucose base extender containing 16% egg yolk and 5% glycerol (Control) or using base extender with no egg yolk, 5% glycerol and 8% low-density lipoprotein (G5L8) with addition of different non-enzymatic antioxidants (10 mM) during 3h of incubation at 38 °C after thawing.

Extenders	Parameters
Extenders	TM (%)	PM (%)	VCL (µm/s)	VSL (µm/s)	VAP (µm/s)	LIN (%)	ALH (µm)
Control	36.1±3.7^a	9.8±1.6^ab	39.7±1.5^a	16.5±1.0^b	22.6±1.1^b	41.0±1.3^ab	2.9±0.1^a
G5L2	34.9±4.3^a	11.2 ± 2.1^a	43.3±1.9^a	18.8±1.4^ab	25.8±1.7^ab	42.6±1.6^ab	2.8±0.1^a
G5L2+AA	34.1±2.9^a	10.1±1.4^ab	42.3±1.7^a	17.2±1.2^ab	23.3±1.3^ab	39.9±1.6^b	3.1±0.1^a
G5L2+HB	35.8±3.3^a	11.8±1.7^a	43.8±2.1^a	20.1±1.5^a	27.2±1.8^a	45.3±2.2^a	2.8±0.1^a
G5L2+AP	5.7±0.6^b	0.0±0.0^c	15.4±0.7^b	2.9±0.3^c	5.9±0.5^c	18.2±1.9^c	0.2±0.1^b
G5L2+TH	28.0±2.5^a	6.9±1.0^bc	38.7±1.8^a	15.4±1.3^b	21.7±1.6^b	38.8±2.0^b	3.1±0.1^a

	Extender	Incubation time
	Extender	0h	1h	2h	3h
TM (%)	Control	36.1 ± 3.7^a	34.3 ± 3.8^ab	27.2 ± 3.1^a	20.4 ± 3.0^ab*
	G5L2	34.9 ± 4.3^a	33.4 ± 3.3^ab	27.1 ± 3.0^a	25.1 ± 3.1^a
	G5L2 + AA	34.1 ± 2.9^a	30.4 ± 2.6^ab	23.6 ± 2.8^a*	19.8 ± 2.3^ab*
	G5L2 + HB	35.8 ± 3.3^a	36.5 ± 3.8^a	25.4 ± 2.1^a*	22.7 ± 2.5^ab*
	G5L2 + AP	5.7 ± 0.6^c	5.2 ± 0.9^c	0.0 ± 0.0^c*	0.0 ± 0.0^c*
	G5L2 + TH	28.0 ± 2.5^ab	26.5 ± 2.5^b	21.3 ± 2.1^a*	17.1 ± 1.4^b*
VCL (µm/s)	Control	39.7 ± 1.5^a	39.9 ± 1.9^a	33.8 ± 2.3^a*	26.7 ± 2.0^b*
	G5L2	43.3 ± 1.9^a	41.1 ± 2.2^a	38.0 ± 2.4^a*	32.4 ± 2.2^a*
	G5L2 + AA	42.3 ± 1.7^a	40.4 ± 2.3^a	33.4 ± 2.0^a*	26.4 ± 1.7^b*
	G5L2 + HB	43.8 ± 2.1^a	44.5 ± 1.8^a	39.5 ± 1.8^a	33.2 ± 2.2^a*
	G5L2 + AP	15.4 ± 0.7^b	14.9 ± 2.0^b	0.0 ± 0.0^b*	0.0 ± 0.0^c*
	G5L2 + TH	38.7 ± 1.8^a	38.4 ± 2.1^a	34.0 ± 2.3^a	30.1 ± 1.6^ab*
LIN (%)	Control	41.0 ± 1.3^ab	36.9 ± 1.2^a	35.8 ± 1.3^a	31.1 ± 1.7^a*
	G5L2	42.6 ± 1.6^ab	34.2 ± 1.2^a	31.2 ± 2.0^ab	29.1 ± 2.3^ab*
	G5L2 + AA	39.9 ± 1.6^b	34.5 ± 1.7^a	30.0 ± 1.5^b	25.5 ± 2.5^bc*
	G5L2 + HB	45.3 ± 2.2^a	35.8 ± 1.3^a	34.7 ± 1.6^a	31.2 ± 2.0^a*
	G5L2 + AP	18.2 ± 1.9^c	12.8 ± 1.7^c	0.0 ± 0.0^c	0.0 ± 0.0^d*
	G5L2 + TH	38.8 ± 2.0^b	29.3 ± 1.4^b	27.9 ± 1.8^b	23.6 ± 1.9^c*

Extenders	Parameters
Extenders	TM (%)	PM (%)	VCL (µm/s)	VSL (µm/s)	VAP (µm/s)	LIN (%)	ALH (µm)
Control	31.9±3.4^c	9.4±1.9^b	36.6±2.3^b	16.0±1.6^c	21.3±1.8^c	42.3±2.0^b	2.7±0.1^a
G5L8	49.5±3.2^a	18.3±1.8^a	48.8±2.0^a	22.6±1.3^ab	30.2±1.5^ab	45.7±1.3^ab	2.8±0.0^a
G5L8 + AA	37.7±3.4^bc	13.4±1.9^ab	47.5±2.0^a	20.8±1.4^ab	27.7±1.6^b	43.3±1.6^b	2.9±0.1^a
G5L8 + HB	46.3±4.6^ab	19.7±3.3^a	50.5±2.4^a	25.1±1.7^a	33.3±2.1^a	49.0±1.7^a	2.7±0.1^a
G5L8 + AP	15.2±0.4^d	0.3±0.1^c	19.8±0.8^c	4.7±0.4^d	9.0±0.6^d	23.6±1.3^c	0.5±0.2^b
G5L8 + TH	36.3±4.5^c	11.5±1.8^b	43.7±2.1^a	18.6±1.5^bc	25.4±1.7^bc	41.6±1.7^b	3.1±0.1^a

	Extender	Incubation time
	Extender	0h	1h	2h	3h
TM (%)	Control	31.9 ± 3.4^c	31.1 ± 3.6^b	24.9 ± 3.8^c	22.9 ± 3.4^c
	G5L8	49.5 ± 3.2^a	41.3 ± 3.7^ab	39.9 ± 3.7^a	34.3 ± 3.7^ab*
	G5L8 + AA	37.7 ± 3.4^bc	36.1 ± 2.9^ab	30.4 ± 2.6^bc	28.0 ± 2.3^bc*
	G5L8 + HB	46.3 ± 4.6^ab	43.5 ± 5.0^a	38.5 ± 4.1^ab	37.3 ± 4.3^a
	G5L8 + AP	15.2 ± 0.4^d	9.2 ± 1.0^c	0.0 ± 0.0^d	0.0 ± 0.0^d*
	G5L8 + TH	36.3 ± 4.5^c	33.3 ± 3.7^ab	25.6 ± 2.6^c	20.6 ± 2.1^c*
VCL (µm/s)	Control	36.6 ± 2.3^b	41.3 ± 2.5^c	32.3 ± 2.2^d	28.8 ± 2.5^b*
	G5L8	48.8 ± 2.0^a	52.9 ± 2.5^ab	50.1 ± 2.2^a	45.5 ± 2.5^a*
	G5L8 + AA	47.5 ± 2.0^a	47.8 ± 2.2^ab	45.0 ± 2.1^ab	41.0 ± 2.5^a*
	G5L8 + HB	50.5 ± 2.4^a	54.4 ± 2.6^a	47.6 ± 2.5^a	47.0 ± 3.1^a
	G5L8 + AP	19.8 ± 0.8^c	16.6 ± 0.5^d	0.0 ± 0.0^e	0.0 ± 0.0^c*
	G5L8 + TH	43.7 ± 2.1^a	45.8 ± 2.5^bc	40.2 ± 2.8^bc	33.0 ± 2.2^b*
LIN (%)	Control	42.3 ± 2.0^b	36.4 ± 1.3^a	33.2 ± 1.9^abc	31.1 ± 2.5^b*
	G5L8	45.7 ± 1.3^ab	35.2 ± 1.2^ab	36.2 ± 1.1^ab	33.9 ± 0.9^ab*
	G5L8 + AA	43.3 ± 1.6^b	37.4 ± 1.3^a	34.4 ± 1.3^abc	35.7 ± 2.1^ab*
	G5L8 + HB	49.0 ± 1.7^a	37.8 ± 1.7^a	37.5 ± 2.0^a	37.1 ± 2.3^a*
	G5L8 + AP	23.6 ± 1.3^d	20.0 ± 1.2^c	0.0 ± 0.0^d	0.0 ± 0.0^c*
	G5L8 + TH	41.6 ± 1.7^bc	34.4 ± 1.3^ab	32.4 ± 1.8^bc	31.3 ± 2.3^b*

[1] #
(In Memoriam)

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Post-thaw quality of ram sperm frozen with different concentrations of low-density lipoproteins associated with non-enzymatic antioxidants

Abstract