Growth And Transcriptomic Response Of Chinese Mitten Crab(Eriocheir Sinensis)to PAHs

Polycyclic aromatic hydrocarbons(PAHs)is a widely spreaded persistent organic pollutant in the water,soil,and air.It has obvious carcinogenic,teratogenic and mutagenic.In order to study the toxic effects of PAHs on crustaceans and transcriptional response,this paper chose the PAHs that has three benzene rings,four benzene rings,and five benzene rings: Phenanthrene(PHE),Chrysene(CHR)and Benzo[α]pyrene(BaP)as the research objects respectively,and Chinsee mitten crab Eriocheir sinensis as experimental animals,which is an important economic aquatic animals in China,PAHs are studied on the toxicity effect of Eriocheir sinensis and transcriptional responses.1.Growth and transcriptomic response of Chinese mitten crab(Eriocheir sinensis)to Phenanthrene(PHE)This study explored the effects of toxicity and transcriptome of low rings PAHs – PHE stressed to Eriocheir sinensis under the safe concentrations(0ug/L,20ug/L,200ug/L)for 28 days,and the 0 ug/L is ethanol control,measured the growth,the response of the transcriptome,detoxification genes’ relative expression and energy metabolism.The results showed that there was no effect on the survival and hepatopancreas index(HSI)under the safe concentrations,but the weight gain and oxygen consumption rate were significantly reduced(P < 0.05).The hepatopancreas transcriptome revealed after 28 day exposure,the energy metabolism pathways and insulin endocrine(ko04910)endocrine signaling pathway and so on has the remarkable difference(P < 0.05),when treatment group compared with controls.In order to further verified the energy metabolism changes,this study also measured the hepatopancreas,glycogen,triglyceride(TGs),total amino acid(T-AA)and lactic acid(LD)content after the PHE stress for 28 days,and the content of serum glucose and LD.The results exhibited that the hepatopancreas glycogen,TGs and T-AA was significantly lower than control group(P < 0.05),however the treatment group of LD content compared with the control group did not show the difference.The content of glucose in the treatment group was significantly lower than that in the control group(P < 0.05),and the LC content decreased with the increase of the concentration of the PHE,but only 200μg/L treatment group showed significant difference compared with the control group(P <0.05).Combined with the transcriptome and energy metabolism index,it indicated that the PHE that under the safe concentrations can cause the crab hormone imbalance,and then disturb the energy metabolism in the Chinese mitten crab.To explore whether there is a AhR-CYP signal pathway in the chinses mitten crab as vertebrate,this study also measured the hepatopancreas genes’ relative expressions of aromatic hydrocarbon receptor(AhR),aromatic receptor nuclear transfer factor(ARNT)and heat shock protein 90(HSP90)after PHE stress for 4 days and 28 days.The results of genes expression showed a good consistency,they were increased in 4 days and 28 days,and exhibited a significant difference in 28 day(P < 0.05),but the dose-effect is not obvious.This indicated that the PHE stress induced the expression of these three detoxification genes,which there may be a similar AhR-CYP xenobiotics biodegradation and metabolism pathway in the chinses mitten crab as the vertebrate.In conclusion,tHSI study showed that under the safe concentration of PHE stress,it can disrupted the chinses mitten crab energy metabolism in the body,in turn,affect the growth,chinses mitten crab may exist the same detoxify animal detoxify metabolism as vertebrate.2.Growth and transcriptomic response of Chinese mitten crab(Eriocheir sinensis)to Chrysene(CHR)This study explored the effects of toxicity and transcriptome of CHR stressed to Eriocheir sinensis under the safe concentrations(0ug/L,10ug/L,100ug/L)for 28 days,and the 0 ug/L is acetone control,measured the growth,the response of the transcriptome,detoxification genes’ relative expression and energy metabolism.The results showed that there was no effect on the HSI,but the survival,weight gain and oxygen consumption rate were significantly reduced(P < 0.05),The hepatopancreas transcriptome revealed after 28 day exposure,the energy metabolism pathways and xenobiotics biodegradation and metabolism pathway has the remarkable difference(P < 0.05),when treatment group compared with controls.In order to further verified the energy metabolism changes,this study also measured the hepatopancreas,glycogen,TGs,T-AA and LC content after the CHR stressed for 28 days,and the content of serum glucose and LD.The results exhibited that the hepatopancreas glycogen,TGs and TAA was significantly lower than control group(P < 0.05),however the LD content compared with the control group did not show the difference.The content of serum glucose in the treatment group was significantly lower than that in the control group(P < 0.05),and the LD content increased with the increase of the concentration of the CHR,and there were significant difference between groups(P <0.05).Combined with the transcriptome and energy metabolism index,it indicated that the CHR disturbed the energy metabolism in the Chinese mitten crab.To explore whether there is an AhR-CYP signal pathway in the Chinese mitten crab as vertebrate,this study also measured the hepatopancreas genes’ relative expressions of AhR,ARNT and HSP90 after CHR stress for 4 days and 28 days.The results of genes expression showed a good consistency,the gene expression in the 10 ug/L group was higher than the control group in 4 days and 28 days,but the 10 ug/L group was no difference compared to the control group.This indicated that the CHR stress induced the expression of these three detoxification genes,which there may be a similar AhR-CYP xenobiotics biodegradation and metabolism pathway in the chinses mitten crab as the vertebrate.It indicated that a certain concentrations of CHR stress can accelerate the energy metabolism in the Eriocheir sinensis,and then influence the growth,and the degradation and metabolic system of foreign substances in the organism be induced.3.Growth and transcriptomic response of Chinese mitten crab(Eriocheir sinensis)to benzo[α]pyrene(BaP)This study mainly to investigate the BaP effects on the toxic and transcriptome.According to the BaP concentrations in the nature water,this study was performed to investigate the effects of BaP on the growth,transcriptomic response and energy metabolism.Eriocheir sinensis exposed to 0.15μg/L or 0.45μg/L BaP for 28 days before sampling.The results exhibited that BaP in the water had significant negative effects on the survival rate,weight gain and oxygen consumption of Eriocheir sinensis,and of the impact were nagetively correlated with BaP concentration.The results of transcriptome analysis showed that the pathways and key enzymes related with energy metabolism significantly effected by BaP.Furthermore,the content of glycogen,TGs and T-AA,in the hepatopancreas were significant lower than that in the control group after the exposure(P <0.05),but no difference observed between different BaP concentrations.The LC in the hepatopancreaswas not effected,while the LC in the serum significantly increased compared with the control group.Serum glucose increased in the exposure group,but only the 0.45μg/L group showed significant difference(P<0.05).Fumarylacetoacetase and GST,which are play a role in the xenobiotics biodegradation and metabolism had higher transcriptional expression in the 0.15μg/L and 0.45μg/L groups than in the control group respectively.It might be indicate that when the Eriocheir sinensis exposed to BaP,the crab can mobilize its immune system and detoxify metabolism to cope with the toxicology effect and eliminate the BaP.In addition,AhR,ARNT and HSP90 in the hepatopancreas also showed higher expression in the two exposure groups than the control group analyzed by RT-PCR,and in the 0.45μg/L group it had a significant different compared to control group at 28d(P <0.05).This indicated that BaP chronic exposure can reduce the growth and survival of Eriocheir sinensis,increased the sugar isogenesis,and produced more ATP to adapt to the adverse environment,and the degradation and metabolic system of foreign substances in the organism also be induced.