1. MATERIALS AND PROCEDURES
1.1 Sample:
The OLE beverages were provided by American Health Product, INC.
1.2 Experimental animals:
Healthy male Kunming mice obtained from the animal breeding unit of Cancer Institute and Hospital of Chinese Academy of Medical Science and PekingUnionMedicalCollege were used for the study.
1.3. Mice Sperm Abnormality Test:
50 sexually matured male mice with 30-50g body weight were selected and randomly assigned to 5 groups. The positive controls were fed with 40mg/kg.b.w. of Cyclophosphamide, negative controls were fed with distilled water; OLE beverages with feeding dose of 1.25 g/kg.b.w., 2.50 g/kg.b.w., and 5.00g/kg.b.w. were given and distilled water was added to prepare the desired concentration. The fed was administered by gavage once a day for 5 days consecutively; then the mice were killed after 30 days after the final feeding. The epididymis was collected to prepare slide with HE staining. 5 mice from each groups was randomly selected, 1000 sperms with complete structure from each selected mice were evaluated and the incidence of abnormal sperm was counted in percentage. Statistical analysis was done
2. RESULT
Mice Sperm Abnormality Test:
In the case of the incidence of sperm abnormality, the table below has shown that there was no significant difference between the OLE-treated group and negative controls (P>0.05), while there was significant difference between the cyclophosphamide-treated group and negative controls (P<0.01). Therefore, the result has shown no sperm abnormality in the OLE- treated groups.
Table 4: The Incidence of Sperm Abnormality in Selected Mice
|
Dosage (g/kg.b.w) |
Animal count |
Sperm count |
Abnormality sperm count |
Incidence of sperm abnormality |
|
|
Control |
0.00 |
5 |
5000 |
86 |
1.72 |
|
Low |
1.25 |
5 |
5000 |
74 |
1.48 |
|
Medium |
2.50 |
5 |
5000 |
84 |
1.68 |
|
High |
5.00 |
5 |
5000 |
76 |
1.52 |
|
40mg/kg.b.w cylophosphamide |
5 |
5000 |
377 |
7.54 |
|
* There were significant differences when compared with the negative controls (P<0.01)
3. SUMMARY
Mice Sperm Abnormality Test: The result was shown to be negative.
XII. The Slide Records of Mice Sperm Abnormality Test
Dosage Group: High (5.00g/kg.b.w.)
|
Abnormal Sperm
Sample No. |
Hookless Sperm |
AmorphousSperm | Banana-shaped head Sperm | Macro cephalic Sperm | Double- headed Sperm | Double- tailed Sperm | Kinked-tailSperm |
Total(%) |
| 1-1 |
4 |
2 |
4 |
3 |
0 |
0 |
0 |
13 |
| 1-2 |
4 |
5 |
3 |
4 |
0 |
0 |
0 |
16 |
| 1-3 |
2 |
12 |
3 |
1 |
0 |
0 |
0 |
18 |
| 1-4 |
3 |
9 |
1 |
0 |
0 |
0 |
0 |
13 |
| 1-5 |
5 |
8 |
3 |
0 |
0 |
0 |
0 |
16 |
| Total |
18 |
36 |
14 |
8 |
0 |
0 |
0 |
76 |
| Sperm Abnormality (%) |
1.8 |
3.6 |
1.4 |
0.8 |
0 |
0 |
0 |
1.52 |
Dosage Group: Medium (2.50g/kg.b.w.)
|
Abnormal Sperm
Sample No. |
Hookless Sperm | AmorphousSperm | Banana-shaped head Sperm | Macro cephalic Sperm | Double -headed Sperm | Double -tailed Sperm | Kinked -tailSperm | Total(%) |
| 2-1 |
1 |
7 |
3 |
1 |
0 |
0 |
0 |
12 |
| 2-2 |
3 |
6 |
4 |
5 |
0 |
0 |
0 |
18 |
| 2-3 |
3 |
11 |
3 |
0 |
0 |
0 |
0 |
17 |
| 2-4 |
5 |
8 |
0 |
0 |
0 |
0 |
0 |
13 |
| 2-5 |
0 |
14 |
10 |
0 |
0 |
0 |
0 |
24 |
| Total |
12 |
46 |
20 |
6 |
0 |
0 |
0 |
84 |
| Sperm Abnormality (%) |
1.2 |
4.6 |
2 |
0.6 |
0 |
0 |
0 |
1.68 |
Dosage Group: Low (1.25g/kg.b.w.)
|
Abnormal Sperm
Sample No. |
Hookless Sperm |
AmorphousSperm | Banana-shaped head Sperm | Macro cephalic Sperm | Double -headed Sperm | Double -tailed Sperm | Kinked -tailSperm |
Total(%) |
| 3-1 |
1 |
7 |
4 |
0 |
0 |
0 |
0 |
12 |
| 3-2 |
1 |
9 |
3 |
1 |
0 |
0 |
0 |
14 |
| 3-3 |
1 |
8 |
7 |
1 |
0 |
0 |
0 |
17 |
| 3-4 |
1 |
10 |
5 |
2 |
0 |
0 |
0 |
19 |
| 3-5 |
1 |
8 |
2 |
1 |
0 |
0 |
0 |
12 |
| Total |
5 |
42 |
21 |
5 |
0 |
0 |
0 |
74 |
| Sperm Abnormality (%) |
0.5 |
4.2 |
2.1 |
0.5 |
0 |
0 |
0 |
1.48 |
Dosage Group: Negative Controls
|
Abnormal Sperm
Sample No. |
Hookless Sperm | AmorphousSperm | Banana-shaped head Sperm | Macro cephalic Sperm | Double -headed Sperm | Double -tailed Sperm | Kinked -tailSperm | Total(%) |
| 4-1 |
4 |
7 |
2 |
0 |
0 |
0 |
0 |
13 |
| 4-2 |
5 |
14 |
1 |
0 |
0 |
0 |
0 |
20 |
| 4-3 |
5 |
5 |
10 |
1 |
0 |
0 |
0 |
21 |
| 4-4 |
2 |
19 |
1 |
1 |
0 |
0 |
0 |
23 |
| 4-5 |
3 |
4 |
2 |
0 |
0 |
0 |
0 |
9 |
| Total |
19 |
49 |
16 |
2 |
0 |
0 |
0 |
86 |
| Sperm Abnormality (%) |
1.9 |
4.9 |
1.6 |
0.2 |
0 |
0 |
0 |
1.72 |
Dosage Group: Cyclophosphamide-treated Group (Positive Controls)
|
Abnormal Sperm
Sample No. |
Hookless Sperm | AmorphousSperm | Banana-shaped head Sperm | Macro cephalic Sperm | Double -headed Sperm | Double -tailed Sperm | Kinked -tailSperm | Total(%) |
| 5-1 | 2 | 62 | 3 | 9 | 0 | 0 | 0 | 76 |
| 5-2 | 5 | 61 | 4 | 6 | 1 | 0 | 1 | 78 |
| 5-3 | 12 | 61 | 4 | 5 | 1 | 0 | 0 | 83 |
| 5-4 | 15 | 40 | 4 | 6 | 0 | 0 | 0 | 65 |
| 5-5 | 5 | 61 | 1 | 7 | 0 | 0 | 1 | 75 |
| Total | 39 | 285 | 16 | 33 | 2 | 0 | 2 | 377 |
| Sperm Abnormality (%) | 3.9 | 28.5 | 1.6 | 3.3 | 0.2 | 0 | 0.2 | 7.54 |
The Incidence of Sperm Abnormality in Selected Mice
|
Dosage (g/kg.b.w) |
Animal Count |
Sperm Count |
Abnormality Sperm Count |
Incidence of Sperm Abnormality (%) |
|
Positive Control |
5 |
5000 |
86 |
1.72 |
|
Low |
5 |
5000 |
74 |
1.48 |
|
Medium |
5 |
5000 |
84 |
1.68 |
|
High |
5 |
5000 |
76 |
1.52 |
|
40mg/kg.b.w cylophosphamide
|
5 |
5000 |
377 |
7.54 |
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