5. SUMMARY AND CONCLUSION
The experimental work of this study was carried out in the Rabbitry Farm of the Department of Animal Production, Faculty of Agriculture, Benha University to improve reproductive performance of male Sinai gabali rabbits during the period from March to November, 2017. This experiment was done on twenty four male Sinai gabali rabbits three months of age, average weight 1.9 ±0.1 kg and divided randomly into four groups each comprised six bucks. The 1st served as control group, the 2nd treated with Wheat germ oil (WGO, 300mg/K B.W.), the 3rd treated with Coenzyme Q10 low dose (CoQ10L 10mg/Kg B.W.) and the 4th treated with Coenzyme Q10 high dose (CoQ10L 20mg/Kg B.W.). Rabbits of all treatments were given oral administration daily for 60 days in the same condition. Fertility experiment carried out on twenty four Sinai gabali dose with average body weight 2.6 ± 0.1kg and six months each were divided into four groups each comprised six rabbits. The 1st group mated via artificial insemination with semen collected from bucks of control group (C). The 2nd group mated via artificial insemination with semen collected from bucks of wheat germ oil (WGO) group. The 3rd group mated via artificial insemination with semen collected from bucks of Coenzyme Q10 low dose (10 mg/kg B.W), the 4th group mated via artificial insemination with semen collected from bucks of Coenzyme Q10 high dose (20 mg/kg B.W).
The objective of this study is to verify the effectiveness of wheat germ oil and the Coenzyme Q10 in low and high doses on physiological performance, semen Quality, DNA migration test (comet assay), oxidative stress markers on testicular cells in male gabali rabbits. In addition to do anatomical sections to study the effect of treatments on the melatonin receptors on the testicular cells in addition to follow the best period of time appropriate under the conditions of Egypt in summer.
Results obtained could be summarized as follows:
A- Blood parameters:
1- Aspartate aminotransferase (AST) U/L:
Treated rabbits with WGO, Coenzyme Q10L and Coenzyme Q10H showed insignificant changes aspartate aminotransferase (AST) activities during the different 6 months. The interactions between treatments and months have no effect and recorded the lowest value of AST (32.72±1.1 U/L) in Coenzyme Q10L×5th month (hot THI) and the highest value 37.33 ± 1.1U/L in control×3rd month (hot THI).
2- Alanine aminotransferase (ALT) U/L:
- Rabbits treated with the daily oral dose of Coenzyme Q10L (10 mg/Kg B.W.) revealed non- significant decreases of ALT value (27.99±0.9 U/L) in comparing with other treatment.
- It was found that, the lowest value of ALT (27.90±1.0 U/L) was in the 6th month of experimental period (October, mild THI), however , the 4th month (August, high THI) showed the highest value (29.70±1.1 U/L) of ALT.
- The results showed non- significant differences between treatments and control on liver enzymes. This means that Coenzyme Q10 and WGO did not show any pathological alteration in liver and indicated the safety for liver enzymatic profile after treatment along 2 months.
3- Protein fractions:
Results did not show any significant differences in blood protein fractions on 4 groups due to the effect of treatments, months and interaction between them.
4- Lipid profile:
- Total cholesterol (TC), triglycerides (TG) and LDL concentrations means show significant decrease (P>0.05) due to WGO and Coenzyme Q10 low and high doses treatments compared with control. Also, serum HDL concentration showed significant increase due to treatment with highest mean of 32.79± 1.1mg/dL for Coenzyme Q10 H group.
- Intervals showed significant changes in TC, TG , LDL and HDL concentration with the highest means of 101.87 ± 3.4 mg/dL in 5th month (Septembre, high THI), 101.81 ± 3.1 mg/dL in 6th month (October, mild THI), 64.12 ± 2.0 mg/dL in 4th month (August, high THI) and 32.81 ± 1.1 mg/dL in 3rd month (July, high THI), respectively.
- Interaction between treatment and months showed a significant (P˂0.0001) differences in TC, TG, LDL and HDL.
A- Reproductive performance:
1-Reaction time (sec) (Libido):
- Effect of treatment on reaction time (sec) showed significant decreas (P>0.05) between groups with the lowest mean of 16.10±0.5 sec. for Coenzyme Q10H treated group.
- Data showed significant differences (P>0.05) between months, and showed the best value of reaction time 16.4±0.6sec.in 6th month of the experimental period (October, mild THI).
- Interaction between treatment and months showed the lowest value of reaction time 15.53±0.6sec. in the Coenzyme Q10H×4th month (August, high THI) and the highest value 18.89±0.6sec. in Coenzyme Q10L×2nd month (June, high THI).
- The results showed that daily oral administration with WGO and Coenzyme Q10 decreases reaction time in Sinai gabali rabbit bucks.
2- Testosterone level (ng/mL):
- Serum testosterone level showed significant (P>0.05) increase due to the effect of Coenzyme Q10L and Coenzyme Q10H with the highest means of 2.95±0.12 ng/mL for Coenzyme Q10H treated group.
- In respect of the effect of months on testosterone level it was found a significant (P>0.05) increase in testosterone concentration between the first 3 months and those of last three months of experimental period (after the end of 60 days of administration the rabbits by WGO and Coenzyme Q10).
- Interaction between treatment and months show significant (P>0.05) differences for testosterone, with the highest mean 3.25±0.1 ng/mL for Coenzyme Q10L×1st month (May, high THI).
3- Ejaculate volume (mL):
- Semen ejaculate volume showed a significant (P˂0.05) increase due to WGO and Coenzyme Q10 (low and high doses) treatment compared with control group with the highest value 0.57 ± 0.02 mL for Coenzyme Q10 H group.
- Interaction between treatment and months of semen samples collection affected the ejaculate volume showed the best value 0.64±0.02 mL in Coenzyme Q10H × 1st month (May, high THI) and the lowest value was 0.42± 0.01 mL in the 2nd month (June, high THI).
4- Sperm cell concentration ×106 /mL:
- Data showed a significant increase (P˂0.05) in sperm cell concentration due to the treatment with Coenzyme Q10H with the highest value 394.02 ± 13.3×106 /mL.
- It was found a significant (P˂0.05) increase in sperm cell concentration means due to different 6 months of experimental duration.
- According to Interaction between treatment and months data showed the highest value 417.00±14.1×106 /mL in Coenzyme Q10H ×2nd month (June, high THI) and the lowest one 367.50±12.2×106 /mL was in WGO ×4th month (August, high THI).
5- Mass motility (grade):
- Means of mass motility showed a significant (P˂0.05) increase due to the treatment with Coenzyme Q10 (low and high doses).
- Data showed a significant (P˂0.05) differences during 6 months of experimental period with the highest grade 3.82±0.1 in the 4th month (high THI).
6- Individual motility (%):
- Means of individual motility showed a significant (P>0.05) increase in treated groups with Coenzyme Q10 compared with other groups.
- It was found non-significant changes due to the effect of months and the interaction between treatment and months.
7- Livability (%):
- Data showed a significant (P>0.05) increase in live sperms percentage due to the effect of Coenzyme Q10 treatment (low and high dose) with the highest mean of 78.79±2.5% for Coenzyme Q10L compared with control group (74.68±2.52 %).
- Analysis of variance showed a significant (P>0.0001) differences in live sperms percentage due to the effect of month and the interaction between month and treatments.
8- Total abnormalities (%):
- Analysis of variance showed non-significant changes in total abnormalities percentage due to the effect of treatment, months and their interaction.
9- Number of services per conception (NSC):
Does inseminated with semen of bucks treated with WGO and Coenzyme Q10 showed a non-significant increase in number of services per conception compared with control group.
10- Conception rate %:
Results showed a significant (P>0.05) increase in conception rate due to the effect of semen of buck treated with WGO, Coenzyme Q10 low and high does with the highest mean 100% for Coenzyme Q10H treated group.
11- Gestation length (day):
Does inseminated with semen of bucks treated with Coenzyme Q10H showed a significant (P>0.05) increase in gestation length with mean 31.3±0.30 day.
C- Productive performance:
1- Litter size at birth (No):
Data showed a significant (P>0.05) increase in litter size at birth due to the effect of semen of buck treated with WGO, Coenzyme Q10 low and high dose compared with control group.
2- Litter size at weaning (No):
Results showed a significant (P>0.05) increase in litter size at weaning due to the effect of semen of buck treated with WGO, Coenzyme Q10 low and high dose compared with control group.
3- Litter weight at birth (g):
Data showed a significant (P>0.05) increase in litter weight at birth due to the effect of semen of buck treated with WGO, Coenzyme Q10 low and high dose compared with control group.
4- Litter weight at weaning (g):
Data showed a significant (P>0.05) increase in litter weight at weaning due to the effect of semen of buck treated with WGO, Coenzyme Q10 low and high dose compared with control group.
5- Mortality rate at weaning (%):
Results of data showed a significant (P˂0.05) decrease in mortality rate due to the effect of semen of buck treated with WGO, Coenzyme Q10 low and high dose compared with control group.
D- Histological study:
1- Oxidative stress markers:
The results showed significant (P˂0.05) decreases in oxidized glutathione (GSSG) and nitric oxide (NO) due to the treatment with WGO and Coenzyme Q10H, while there was a non- significant increase in testes tissues for bucks treated with Coenzyme Q10L. Also this study showed a significant (P˂0.05) decrease in testes tissue oxidative stress marker malondialdehyde (MDA) and increase reduced glutathione (GSH) that may be due to Coenzyme Q10 and WGO antioxidant activity.
2- Identification testicular expression of melatonin receptors MT1 and MT2:
- The melatonin receptor was highly expressed in testicular cells in Coenzyme Q10H group and melatonin has potent effect on testicular development.
3- Single- cell gel electrophoresis assay (Comet assay):
Data showed insignificant changes in cell head and significant (P˂0.05) increase in DNA head and a significant (P˂0.05) decrease in Tail, DNA tail and Tail moment.
From the previous results, it could be concluded that, the WGO and Coenzyme Q10 have powerful effect on physiological and reproductive traits on males Sinai gabali rabbits compared with control group. Especially Coenzyme Q10H was the best treatment compared with the others. The results of the present study indicate that administration of WGO and Coenzyme Q10 reduced total cholesterol, triglycerides, low density lipoprotein and increased high density lipoprotein, also the present study suggests that WGO and Coenzyme Q10 treatments improve physiological and biochemical performance, semen quality, reduced heat stress and testicular oxidative stress markers in Sinai gabali male rabbits under Egyptian summer condition.
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