Enhancing semen quality, brain neurotransmitters, and antioxidant status of rabbits under heat stress by acacia gum, vitamin C, and lycopene as dietary supplements: an in vitro and in silico study

Figures & data

Table 1. Composition and calculated analyses of the basal diet (g/kg, as-fed basis) of weaned rabbits (Atallah et al. 2021).

Ingredients	Content
Ingredients, g/kg
Alfalfa hay	350.0
Yellow corn	200.0
Soybean meal	96.0
Wheat bran	300.0
Corn Stover	30.0
Di-calcium phosphate	12.5
L-Lysine HCl	2.1
DL-Methionine	2.0
Sodium chloride	5.0
Vitamin/mineral premix*	1.5
Total	1000.0
Calculated analysis, g/kg on dry matter basis
Digestible energy, MJ/kg	11.6
Crude protein, g/kg	179.0
Crude fibre, g/kg	125.0
Crude fat, g/kg	32.0
Ca, g/kg	10.9
Available P, g/kg	5.9
Methionine, g/kg	4.2
Lysine, g/kg	9.0

*Minerals and vitamins mixture supply/kg of diet: vit. A, 20,000 IU; vit. D3, 15,000 IU; vit. E, 8.33 g; vit. K, 0.33 g; vit. B1, 0.33 g; vit. B2, 1.0 g; vit. B6, 0.33 g; vit. B5, 8.33 g; vit. B12, 1.7 mg; pantothenic acid,3.33 g; biotin, 33 mg; folic acid, 0.83 g; choline chloride, 200 g.

Table 2. Average values of air temperature (T), relative humidity (RH), and temperature humidity index (THI) at the experimental site.

Measurements	Period, days
	15	30	45	60	75	90
T, db°C	30.20	32.6	33.40	34.00	34.4	35.90
RH%	62.30	63.1	64.20	64.70	65.1	65.80
THI	28.30	30.5	31.31	32.04	32.4	33.75

T, db°C: dry bulb temperature in degrees Celsius; RH: relative humidity percentage; THI: relative temperature humidity index.

Figure 1. HPLC chromatogram of Acacia gum.

Table 3. Nutritional impacts of acacia gum, vitamin C, and lycopene on semen quality New Zealand rabbits under heat stress.

Treatment	Semen quality
	Sperm motility	Motility index	Sperm normality, %	Sperm abnormality, %	Live sperm, %	Dead sperm, %
Control	2.28^c	48.06^c	57.09^c	43.08^a	59.60^c	37.66^a
Vitamin C	3.61^ab	64.66^b	72.17^b	28.41^c	64.94^b	33.14^b
Lycopene	3.34^b	59.12^b	62.95^c	36.85^b	63.85^bc	34.38^b
Acacia gum	3.87^a	71.31^a	80.13^a	17.70^d	94.15^a	9.33^c
SEM	0.07	1.45	1.63	0.73	1.29	0.69
p-Value	0.0001	0.0001	0.0001	0.0001	0.0001	0.0001

^a–dMean values within a column with different superscript letters are significantly different.

Figure 2. Nutritional impacts of acacia gum, vitamin C and lycopene on brain neurotransmitter New Zealand rabbits under heat stress. ^a–dMean values within the same box with different superscript letters are significantly different. p < 0.001.

Table 4. Nutritional impacts of acacia gum, vitamin C, and lycopene on semen antioxidants New Zealand rabbits under heat stress.

	Semen
Treatment	GSH, μmol/mL	TAC	SOD, Ul/mµL	CAT, μmol of H2O2/mL	L-Car	Na^+-K^+ ATPase, nmol/g tissue	ATP	Total calcium, mmol/L	MDA, nmol/mL	NO
Control	0.34^c	19.90^b	36.08^c	13.45^c	315.18^c	402.01^b	0.30^c	2.98^b	38.79^a	0.67^a
Vitamin C	0.61^a	26.04^a	44.16^a	17.23^ab	344.65^ab	467.23^a	0.54^b	3.49^a	31.74^b	0.45^c
Lycopene	0.48^b	22.19^b	40.94^c	16.58^b	324.36^bc	403.55^b	0.54^b	3.19^ab	18.58^c	0.52^b
Acacia gum	0.64^a	27.20^a	46.13^b	18.15^a	372.80^a	439.48^a	0.63^a	3.41^a	14.01^d	0.41^c
SEM	0.01	0.63	0.78	0.38	7.12	10.76	0.01	0.08	0.71	0.01
p-Value	0.0001	0.0001	0.0001	0.0001	0.0001	0.0001	0.0001	0.0009	0.0001	0.0001

^a–dMean values within a column with different superscript letters are significantly different.

Figure 3. The aromatic interaction and the 2D chemical interactions of the docked complexes between catalase enzyme and (A,B) lycopene, (C,D) vitamin-C, (E,F) phosphatidylserine, and (G,H) phosphatidylcholine.

Figure 4. The aromatic interaction and the 2D chemical interactions of the docked complexes between glutathione peroxidase enzyme and (A,B) lycopene, (C,D) vitamin-C, (E,F) phosphatidylserine, and (G,H) phosphatidylcholine.

Figure 5. The aromatic interaction and the 2D chemical interactions of the docked complexes between superoxide dismutase enzyme and (A,B) lycopene, (C,D) vitamin-C, (E,F) phosphatidylserine, and (G,H) phosphatidylcholine.

Figure 6. The H-bond acceptor in green and doner in pink colours, charges positive with blue and negative with red colours, a hydrophobic area with a brown and hydrophilic area with blue colours, ionizability with basic in blue and acidic in red colours, and solvent accessible surface (SAS) in blue colour through the interactions of the docked complexes between catalase enzyme and (A–E) lycopene, (F–J) vitamin-C, (K–O) phosphatidylserine, and (P–T) phosphatidylcholine, respectively.

Figure 7. The H-bond acceptor in green and doner in pink colours, charges positive with blue and negative with red colours, a hydrophobic area with a brown and hydrophilic area with blue colours, ionizability with basic in blue and acidic in red colours, and solvent accessible surface (SAS) in blue colour through the interactions of the docked complexes between glutathione peroxidase enzyme and (A–E) lycopene, (F–J) vitamin-C, (K–O) phosphatidylserine, and (P–T) phosphatidylcholine, respectively.

Figure 8. The H-bond acceptor in green and doner in pink colours, charges positive with blue and negative with red colours, a hydrophobic area with a brown and hydrophilic area with blue colours, ionizability with basic in blue and acidic in red colours, and solvent accessible surface (SAS) in blue colour through the interactions of the docked complexes between superoxide dismutase enzyme and (A–E) lycopene, (F–J) vitamin-C, (K–O) phosphatidylserine, and (P–T) phosphatidylcholine, respectively.

Table 5. The docking analysis results of binding energy, No. of H-bond, No. of carbon H-bond, hydrophobicity, and SAS interactions between GSH, SOD, and CAT enzymes against vitamin C, lycopene, phosphatidylcholine, and phosphatidylserine.

Component	Binding energy			No. of conventional H-bond			No. of carbon H-bond			Hydrophobicity			SAS
	GSH	SOD	CAT	GSH	SOD	CAT	GSH	SOD	CAT	GSH	SOD	CAT	GSH	SOD	CAT
Vitamin C	−5.1	−5.8	−6.6	3	3	4	0	0	0	High	High	Moderate	Moderate	High	Moderate
Lycopene	−6.4	−7.1	−8.6	0	0	0	0	0	0	High	High	Moderate	High	Moderate	Low
Phosphatidylcholine	−4.2	−4.3	−6.9	3	3	1	3	1	0	High	High	High	High	Moderate	High
Phosphatidylserine	−4.4	−5.0	−5.2	4	3	4	0	3	1	Low	High	Moderate	High	High	Moderate

GSH: glutathione peroxidase; TAC: antioxidant capacity; SOD: superoxide dismutase; CAT: catalase; L-Car: L-Carnitine; ATP: adenosine triphosphate; MDA: malondialdehyde; NO: nitric oxide.

Atallah AA, Osman A, Sitohy M, Gemiel DG, El-Garhy OH, Azab IHE, Fahim NH, Abdelmoniem AM, Mehana AE, Imbabi TA. 2021. Physiological performance of rabbits administered buffalo milk yogurts enriched with whey protein concentrate, calcium caseinate or Spirulina platensis. Foods. 10(10):2493.

 PubMed Web of Science ®Google Scholar