Effects Of Zearalenone And Its Degrading Enzymes On Reproductive Performance Of Sows

Zearalenone(ZEN)is a kind of secondary metabolites which is teratogenic carcinogenic mainly produced by Fusarium graminearum,can contaminate many economically important crops,resulting in serious food and feed safety problems all over the world.zearalenone(ZEN)is a nonsteroidal oestrogenic mycotoxin mainly produced by a variety of Fusarium fungi and associated with hyperestrogenism and other severe livestock feed and human food with levels in the range of ppb and low mg/kg.It was hypothesized that ZEA,an endocrinedisruptor,could affect puberty and early pregnancy.,leading to numerous economic losses and great health risks all over the world every year.Increasing attention has been paid to the development of effective and efficient strategies of ZEN detoxification.How to treat safely and effectively the contaminated food or feed has always been a thorny issues.Traditional physical and chemical methods of detoxification are in poor operability,easy to destroy the food nutrients and to introduce the secondary pollution.The biological degradation methods are favored because of its advantages such as mild reaction conditions,no harmful reagents residue.The purpose of this study was to establish a expressing system of ZEN degradation enzyme gene(ZEN-jjm)in bacillus subtilis,determine biodegradation activity and protection effect on reproductive performance of sows of ZEN-jjm expressed by bacillus subtilis.This experimental method The recombinant expression plasmid was constructed by ligation cloned ZEN-jjm and pHT01 vector digested with EcoRⅠand NotⅠ,and then was transformed into Bacillus subilis.Expression of ZEN-jjm in Bacillus subilis was analyzed by SDS-PAGE.The activity degrading ZEN of ZEN-jjm expressed in Bacillus subilis was determined by HPLC.The protection effect on reproductive performance of sows of ZEN-jjm expressed by bacillus subtilis was evaluated through the total number of litter size,number of stillbirths,weak young,piglet birth weight.The results of double enzyme digestion and DNA sequencing demonstrated that ZEN-jjm was inserted into pHT01.The SDS-PAGE showed that one Bacillus subilis strain with high-level expression was obtained,and the size of the expressed protein was about 29 kDa.The HPLC result illustrated that the expressed ZEN-jjm protein could effectively degrade ZEN.The evaluation results of reproductive performance of sows indicated that the expressed ZEN-jjm protein could effectively relieve the toxicity of sows caused by ZEN.Conclusion: 1.Sow gestation fodder to add a low dose of ZEN(1.5 mg/kg)pollution can significantly increase the sow stillbirth number and incidence of weak piglets,this may be related to ZEN by disrupting the endocrine system,reproductive system caused by developmental disorder,sow the placenta fetal congenital immune suppression;At the same time,the fodder in the pollution of low dose of ZEN during pregnancy will cause the sow low immunity,reduce the sow fertility.2.The ZEN-jjm expression vector was successfully constructed and the ZEN-jjm protein was expressed in bacillus subtilis.The expressed ZEN-jjm had the biological activity degrading ZEN.3.ZEN-jjm could significantly reduce the harm of ZEN on reproductive performance of sows when it was added in the feed.