الفهرس 
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Abstract
Contamination by Aflatoxin is generally considered to be a problem in tropical and subtropical regions of Asia, Africa and Latin America, Aflatoxin B1 is the most potent natural carcinogen known. Low concentrations of the Aflatoxins, especially AFB1 can affect male reproduction, spermatogenesis, Leydig cell function and fertility.
This study aimed to investigate the effect of Aflatoxin-B1 on male reproductive system clinically on human and experimentally on rabbits.
It was conducted on 100 male attending outpatients Andrology Clinic of Hospital in Sohag University. Patients were classified according to their seminogram into two equal groups. The first one had normal seminogram, while the second had abnormal seminogram. Quantitative analysis for seminal plasma AFB1 was done for both groups.
With regard to the experimental animal study, it lasted for 63 days to investigate effects of AFB1. A total number of 45 NZW rabbit bucks were divided into three equal groups. Each group was divided into equal three subgroups; the 1st subgroup was considered as negative control, while the 2nd and 3rd were positive control and treatment subgroup, respectively.
Rabbit bucks in negative control received the commercial rabbit ration only, while the bucks in positive control received the vehicle by i.p. injection. Finally, bucks in treated subgroups daily received Aflatoxin B1 by i.p injection at 20 µg/ kgBW for different durations according to the group.
The first group, treated rabbits received Aflatoxin B1 by i.p. injection at 20 µg/ kgBW per day for 30 day. The second group, treated rabbits received Aflatoxin B1 by i.p. injection at 20 µg/ kgBW per day for 48 day. The third group, treated rabbits received Aflatoxin B1 by i.p. injection at 20 µg/ kgBW per day for 63 day.
At the end of the study, rabbits were sacrificed where the testis and epididymis removed for histopathological examination. Quantitative analysis for Aflatoxin B1 for animals was done by HPLC- Flourescent detector.
The data obtained from both human and animal study were analyzed statistically using SPSS version (15.0).
Human study:
The present case-control work showed no significant statistical difference in the mean value of age and seminal fluid volume between the first and second human groups. There was significant decrease in the mean value of sperm concentration per ml (×106) and the percentage of progressive sperm motility and total sperm motility in second group as compared with the first human groups. On contrary, there was significant increase in the mean value of the percentage of abnormal sperm forms, and seminal plasma AFB1 level (pg/ml) in second group as compared with the first human groups.
The current study showed no significant correlation between seminal plasma AFB1 level (pg/ml) and seminal fluid volume. There were significant negative correlations between seminal plasma AFB1 level (pg/ml) & sperm concentration per ml (×106) and the percentage of progressive sperm motility and total sperm motility. On the other hand there were significant positive correlations between seminal plasma AFB1 level (pg/ml) and the percentage of abnormal sperm forms.
Experimental study:
As regard the rabbit bucks in the first group, there was no significant statistical difference in the mean value of seminal fluid volume (ml), sperm concentration per ml (×106) and the percentage of sperm viability, sperm motility and abnormal sperm forms in the post-treatment rabbits as compared with the negative control, positive control and pre-treatment rabbits inspite of significant increase in the mean value of seminal plasma AFB1 level (pg/ml) in post-treatment rabbits as compared with negative control, positive control and pre-treatment rabbits.
In the second group, the findings showed no statistical difference in the mean value of seminal fluid volume (ml), sperm concentration per ml (×106) and the percentage of abnormal sperm forms for rabbit bucks in the post-treatment rabbits as compared with the negative control, positive control and pre-treatment rabbits inspite of significant increase in the mean value of seminal plasma AFB1 level (pg/ml) in post-treatment rabbits as compared with the negative control, positive control and pre-treatment rabbits.
These results found that the mean value of the percentage of sperm viability and total sperm motility of the second group rabbit bucks showed no significant statistical difference in the post-treatment rabbits as compared with the positive control and pre-treatment rabbits but showed significant decrease as compared with the negative control.
The results in the third group showed no significance difference in the mean value of seminal fluid volume (ml) for rabbit bucks in the post-treatment rabbits as compared with the negative control, positive control and pre-treatment rabbits.
On contrary, the results in the third group for rabbit bucks showed significant decrease in the mean value of sperm concentration per ml (×106) and the percentage of sperm viability and total sperm motility in the post-treatment rabbits as compared with negative control, positive control and pre-treatment rabbits. However, there was a significant increase in the mean value of seminal plasma AFB1 level (pg/ml) and the percentage of abnormal sperm forms of the post-treatment rabbits as compared with the negative and positive control groups as well as pre-treatment rabbits.
In this study, the results showed no significant difference in the mean value of seminal fluid volume between the first & second group treated rabbits, the first & third group treated rabbits, the second & third group treated rabbits nor different treated rabbit groups.
Referring to the mean value of sperm concentration per ml (×106)and the percentage of sperm viability and total sperm motility, there was no significant statistical difference between the first & second group treated rabbits but there was significant decrease between the first & third group treated rabbits, the second & third group treated rabbits and different treated rabbit groups.
As regard the mean value of the percentage of abnormal sperm forms, there was no significant difference between the first & second group treated rabbits, but there was significant increase between the first & third group treated rabbits, the second & third group treated rabbits and different treated rabbit groups.
The mean value of seminal plasma AFB1 (pg/ml), there was significant difference between the first & second group treated rabbits, the first & third group treated rabbits, the second & third group treated rabbits and different treated rabbit groups.
Referring to correlation, the results showed no significant correlation between the seminal AFB1 level and seminal fluid volume. On the other hand the findings showed significant negative correlation between the seminal AFB1 level and [sperm concentration per ml (×106) and the percentage of sperm viability and total sperm motility] but showed significant positive correlation with the percentage of abnormal sperm forms.
The results in the first, second and third rabbit groups showed no significant difference in the mean value of the percentage of seminiferous tubules area between negative and positive control groups, while there was significant decrease in the mean value of the percentage of seminiferous tubules area in the treated rabbits as compared with the negative and positive control groups. On the other hand there was no significant difference in the mean value of the percentage of interstitial area between the negative and positive control groups, but there was significant increase in the mean value of the percentage of interstitial area in the treated rabbits as compared with the negative and positive control groups.
Referring to the mean value of the percentage of seminiferous tubules area, the findings showed significant decrease between the first & second group treated rabbits, the first & third group treated rabbits, the second & third group treated rabbits and different rabbit groups. On the other hand, there was significant increase in the mean value of the percentage of interstitial area between the first & second group treated rabbits, the first & third group treated rabbits, the second & third group treated rabbits and different rabbit groups.
In the present study histopathological examination showed degeneration of seminifrous tubules, which increased by increased duration of treatment. The tubules showed absence of mature sperms with appearance of uninucleated and multinucleated giant cells. Epididymal epithelium showed vaculation and degeneration which increased by increased duration of treatment.
Conclusion:
from the obtained results, it has been found that Aflatoxin B1 affect male reproductive system functionally and pathologically, which increase with increasing exposure duration.
Recommendatiom:
It is advised to:
1. Measure Aflatoxin B1 level in seminal plasma of infertile male as one of the causes of male infertility.
2. Screening for Aflatoxin B1 in food and animal feed and its metabolite in milk (M1) in Sohag Governorate to determine the extent of the problem.