Nano green tea extract (NGTE) in a Tris-soybean lecithin extender improves post-thaw ram sperm motility, kinematics and antioxidant status

Figures (2) Tables (4)

Figure 1.
The physicochemical characteristics of the synthesized NGTE were analyzed using transmission electron microscopy (TEM) and dynamic light scattering (DLS). (a) The TEM image shows the morphology and particle distribution, while the (b) DLS profile presents the log-normal particle size distribution.
Figure 2.
Antioxidant indices and enzyme activities (TAC, MDA, AST, ALT, LDH) in post-thaw sperm medium with NGTE. NGTE = nano green tea extract; TAC = total antioxidant capacity; MDA = malondialdehyde; LDH = lactate dehydrogenase; AST = aspartate aminotransferase; ALT = alanine aminotransferase.

	Items	Control	NGTE_{50 μg/mL}	NGTE_{100 μg/mL}	NGTE_{150 μg/mL}	NGTE_{200 μg/mL}
Post-equilibration	PM	75.30 ± 0.30^e	84.80 ± 0.24^b	85.70 ± 0.26^a	82.40 ± 0.26^c	80.10 ± 0.23^d
	livb	76.30 ± 0.33^e	85.90 ± 0.23^b	86.70 ± 0.26^a	83.50 ± 0.26^c	81.20 ± 0.24^d
	MbinT	76.50 ± 0.26^d	86.80 ± 0.32^a	87.10 ± 0.31^a	84.20 ± 0.29^b	82.20 ± 0.35^c
	Abn	7.70 ± 0.26^c	7.70 ± 0.15^c	7.30 ± 0.15^c	9.00 ± 0.2^b	10.10 ± 0.31^a
Post-thawing	PM	41.50 ± 0.22^c	43.20 ± 0.32^b	46.30 ± 0.21^a	43.90 ± 0.27^b	40.60 ± 0.22^d
	livb	42.80 ± 0.24^c	44.20 ± 0.29^b	47.10 ± 0.27^a	44.80 ± 0.29^b	41.70 ± 0.21^d
	MbinT	43.90 ± 0.27^c	44.80 ± 0.32^c	47.60 ± 0.26^a	46.10 ± 0.31^b	42.50 ± 0.40^d
	Abn	20.70 ± 0.26^b	17.10 ± 0.34^c	16.30 ± 0.15^d	21.10 ± 0.23^b	22.70 ± 0.33^a
Values are presented as mean ± SE. Means within a row followed by different superscripts (a–e) differ significantly (p < 0.05). Abbreviations: NGTE = nano green tea extract; PM = progressive motility; Livb = livability; MbinT = membrane integrity; Abn = abnormality.

Table 1.

Post-equilibration and post-thaw sperm viability, membrane integrity, and abnormality (%) after NGTE supplementation.

Item	Control	NGTE_{50 μg/mL}	NGTE_{100 μg/mL}	NGTE_{150 μg/mL}	NGTE_{200 μg/mL}
RPV	25.40 ± 0.63^c	27.50 ± 0.65^b	33.80 ± 0.53^a	27.20 ± 0.61^b	22.50 ± 0.26^d
SPV	7.30 ± 0.55^a	7.40 ± 0.45^a	4.90 ± 0.31^b	7.90 ± 0.31^a	8.30 ± 0.26^a
TPV	32.70 ± 0.36^c	34.90 ± 0.37^b	38.70 ± 0.39^a	35.10 ± 0.31^b	30.80 ± 0.20^d
NPV	6.80 ± 0.24^b	6.30 ± 0.15^b	5.60 ± 0.26^c	6.80 ± 0.20^b	7.80 ± 0.20^a
TMY	39.50 ± 0.22^c	41.20 ± 0.32^b	44.30 ± 0.21^a	41.90 ± 0.27^b	38.60 ± 0.30^d
IMM	60.50 ± 0.22^b	58.80 ± 0.32^c	55.70 ± 0.21^d	58.10 ± 0.27^c	61.40 ± 0.22^a
Values are presented as mean ± SE. Means within a row followed by different superscripts (a–e) differ significantly (p < 0.05). Abbreviations: NGTE = nano green tea extract; RPV = rapid progressive; SPV = slow progressive; TPV = total progressive; NPV = non-progressive; TMY = total motility; IMM = immotile.

Table 2.

Distribution of motility fractions (%) and progressive motility in frozen–thawed ram semen with NGTE.

Item	Control	NGTE_{50 μg/mL}	NGTE_{100 μg/mL}	NGTE_{150 μg/mL}	NGTE_{200 μg/mL}
DAP	16.60 ± 0.22^d	20.20 ± 0.20^b	27.20 ± 0.20^a	19.70 ± 0.21^b	17.60±0.26^c
DCL	24.40 ± 0.37^e	28.80 ± 0.20^c	29.80 ± 0.29^b	34.00 ± 0.33^a	26.30±0.21^d
DSL	11.40 ± 0.22^d	13.30 ± 0.15^b	13.40 ± 0.22^b	15.70 ± 0.15^a	12.40±0.16^c
VAP	40.00 ± 0.33^d	46.20 ± 0.24^b	46.10 ± 0.23^b	54.80 ± 1.01^a	43.80±0.46^c
VCL	57.90 ± 0.23^d	72.90 ± 0.23^b	72.70 ± 0.33^b	82.60 ± 0.22^a	63.10±0.34^c
VSL	24.90 ± 0.23^d	31.80 ± 0.20^b	30.60 ± 0.26^c	34.70 ± 0.36^a	32.10±0.37^b
STR	62.26 ± 0.50^c	68.84 ± 0.47^b	66.39 ± 0.65^b	63.32 ± 1.5^c	73.32±0.89^a
LIN	43.01 ± 0.43^bc	43.62 ± 0.20^b	42.09 ± 0.42^c	42.01 ± 0.47^c	50.89±0.72^a
WOB	69.09 ± 0.67^a	63.37 ± 0.31^c	63.42 ± 0.39^c	66.35 ± 1.28^b	69.44±0.92^a
ALH	4.50 ± 0.16^b	5.20 ± 0.20^a	5.00 ± 0.21^ab	3.70 ± 0.15^c	4.49±0.17^b
BCF	14.80 ± 0.24^c	18.20 ± 0.24^b	17.80 ± 0.24^b	23.30 ± 0.33^a	23.10±0.27^a
Values are presented as mean ± SE. Means within a row followed by different superscripts (a–e) differ significantly (p < 0.05). Abbreviations: CASA = computer-assisted sperm analysis; NGTE = nano green tea extract; DAP = distance average path (µm); DCL = distance curved line (µm); DSL = distance straight line (µm); VAP = velocity average path (µm/s); VCL = velocity curved line (µm/s); VSL = velocity straight line (µm/s); STR = straightness (VSL/VAP, %); LIN = linearity (VSL/VCL, %); WOB = wobble (VAP/VCL, %); ALH = amplitude of lateral head displacement (µm); BCF = beat cross frequency (Hz).

Table 3.

CASA kinematic parameters of post-thaw ram sperm with NGTE.

Item	Control	NGTE_{50 μg/mL}	NGTE_{100 μg/mL}	NGTE_{150 μg/mL}	NGTE_{200 μg/mL}
NFm (%)	77.3 ± 0.26^c	80.9 ± 0.34^b	81.7 ± 0.15^a	76.9 ± 0.23^c	75.3 ± 0.33^d
AFm (%)	22.7 ± 0.26^b	19.1 ± 0.34^c	18.3 ± 0.15^d	23.1 ± 0.23^b	24.7 ± 0.33^a
NAb (%)	7.3 ± 0.53^b	6.5 ± 0.22^c	6.7 ± 0.26^c	8.2 ± 0.29^ab	8.6 ± 0.40^a
HAb (%)	5.7 ± 0.30^c	6.8 ± 0.20^a	6.7 ± 0.21^a	8.4 ± 0.34^a	8.5 ± 0.40^a
TAb (%)	9.7 ± 0.74^a	5.8 ± 0.24^cd	4.9 ± 0.43^d	6.5 ± 0.52^bc	7.6 ± 0.49^b
Values are expressed as mean ± SE. Means within a row followed by different superscripts (a–e) indicate significant differences (p < 0.05). Abbreviations: NGTE = nano green tea extract; NFm = normal forms; AFm = abnormal forms; NAb = neck abnormalities; HAb = head abnormalities; TAb = tail abnormalities.

Table 4.

Sperm morphology: normal/abnormal forms and head/neck/tail abnormalities (%) after NGTE treatment.