Effects Of Butylbenzyl Phthalate And Triclosan On The Growth And Development Of Adults Of Pseudomonas Black Spiny Ants

Butyl benzyl phthalate（BBP）and Triclosan（TCS）are two kinds of additives often used in daily necessities.BBP and TCS can be detected in water and soil.The various chemical pollutants of the environment can endanger the survival,reproduction,growth,and development of organisms and break the balance of ecosystem.Studies have shown that exposure to BBP and TCS can disturb the hormone level of endocrine system which play crucial roles in the growth and development of organisms.These two major anthropogenic pollutants effecting mammals and aquatic economic species has been getting lot of attention.Few studies are focused on the terrestrial species,especially in the insects.In addition to being a digestive organ,the intestines of animals also participate in endocrine and metabolic activities.A good deal of microorganism exists in the gut and participate in the whole life cycle of the host directly or indirectly.However,the composition of intestinal microflora is diverse.It is easily affected by environment,diet,and other factors,resulting in changes in the diversity of intestinal microflora,and endangering the host health.Insects are the largest animal group on land.It is very important to study the effects of BBP and TCS on the development of insects and the structure and abundance of intestinal flora for exploring the mechanism of exogenous compounds interfering with the growth and development of organisms.The adult of Polyrhachis vicina Roger was selected as the research object.Morphological observation,paraffin section,H＆E staining and qRT-PCR and 16S high-throughput sequencing technology were used to research the effects of BBP and TCS exposure on the growth and development of P.vicina adults,and the effects of TCS exposure on the structure and abundance of intestinal microflora of workers.The effects of BBP and TCS on the growth and development of social insects were further discussed.The research results are as follows:1.Workers were exposed to 0.2 mg/l,2 mg/l,20 mg/l,200mg/l,and 2g/l BBP,and females and males were exposed to 0.2 mg/l,2 mg/l,and 20mg/l BBP after abdominal administration for 15 days.Compared with the control group,the survival rate of workers was reduced after BBP treatment,and the survival rate of males was reduced in a dose-dependent way,but the survival rate of females was not affected.After BBP treatment,the body weight of worker and male ants increased,while the body weight of female ants decreased.2.Compared with the control group,appearance observation showed that the abdomen of different castes of P.vinica treated with BBP had abnormal epidermal growth.The results showed that BBP had direct effect on the normal growth and development of P.vinica.3.The changes of intestinal tissue of workers treated with BBP were observed by paraffin section and hematoxylin eosin staining.The results showed that compared with the control group,BBP treatment mainly affected the midgut of workers.In BBP group,the boundary of midgut cells was not clear,columnar cells degenerated,cytoplasm was damaged,and peritrophic membrane disappeared.It is suggested that BBP may be absorbed by digestive organs and entered various tissues and cells,causing ant poisoning,and destroying its morphological structure.4.qRT-PCR was used to detect the changes of mRNA expression levels of growth and development related genes in different castes of P.vinica.Compared with the control group,the results showed that the mRNA expression levels of PvERR,PvEcR,PvUSP,PvTH,PvFoxO and PvTOR were up-regulated and down regulated in different castes of P.vinica after BBP treated but there were no significant time-dependent and dose-dependent changes.After BBP exposure,the expression trends of PvEcR and PvUSP mRNA in different castes of adults were similar.5.The workers were exposed to 0.02 mg/l,0.2 mg/l,2 mg/l,20 mg/l and 200mg/l TCS,and females and males were exposed to 0.02 mg/l,0.2 mg/l and 2mg/l TCS.The survival rate of workers was reduced and the survival rate of female and male ants decreased in dose-dependent manner.After TCS exposure,workers’ weight increased,female decreased,and male weight had no effect.6.The results of appearance comparison showed that the abdomen of different castes of P.vinica treated by TCS was abnormal.The results showed that TCS had a direct effect on the normal growth and development of P.vinica.7.Paraffin section and hematoxylin-eosin staining were used to observe the changes of intestinal tissue of workers.The results showed that TCS mainly affected the midgut in the workers.In TCS treated group,the muscle layer was incomplete,the cell boundary was unclear,the epithelial cells degenerated,the cytoplasm was damaged,the nucleus pyknosis even disappeared,and the microvilli disappeared.It is inferred that TCS is absorbed by digestive organs and enters various tissues and cells,causing ant poisoning and destroying its morphological structure.8.The expression of TCS related genes in d different castes of P.vinica was detected by qRT-PCR.The results showed that the mRNA expression levels of PvERR,PvEcR,PvUSP,PvTH,PvFoxO and PvTOR in different castes of P.vinica in the treatment group were up-regulated and down regulated compared with the control group,but there were no significant time-dependent and dose-dependent changes.After TCS exposure,the expression trends of PvEcR and PvUSP mRNA in different castes of adults were similar.9.16S high-throughput sequencing technology was used to analyze the effect of TCS on the intestinal microflora of workers.The results showed that TCS can change the richness and composition of the intestinal microbiota of workers,affect the metabolism in vivo,and adversely affect the health of workers.In conclusion,BBP and TCS can cause the death,body weight change,deformity,intestinal tissue structure change and gene expression change closely related to growth and development of P.vinica.In addition,TCS exposure can change the intestinal microbial diversity,structure,and composition of workers,affect the metabolic level,and increase the risk of disease.These findings may provide a new idea and experimental material for evaluating insect response to BBP and TCS as environmental endocrine disruptors.