Insecticidal Potential Of Eichhornia Crassipes Extracts On Agrotis Ipsilon And Its Safety Assessment Against ALBINO Rats

Insect pests pose a serious threat and cause substantial damage to the agricultural sector.The black cutworm Agrotis ipsilon（Hufnagel）（Lepidoptera:Noctuidae）is one of the major pests with a wide variety of economically important crop hosts.This underground cutworm significantly damaged different species of crops,including agronomic,horticultural,and ornamental crops.Usually,chemically synthesized insecticides,such as chlorantraniliprole,chlorpyrifos,deltamethrin,etc.,have been used widely to control this pest.While the unscientific use of insecticides might lead to the development of insecticide resistance to pests and accumulating pesticide residues in food.Hence,it is becoming clear that alternative pest control strategies are required to limit hazards to humans,the environment,and non-target species.The water hyacinth Eichhornia crassipes（Mart.）Solms（Commelinales:Pontederiaceae）is one of the most destructive free-floating aquatic weeds;a single plant may generate over 48,000 new plants monthly.Moreover,water hyacinth uses around three billion m3 annually,which could irrigate approximately 404685.6 m² of fields.Thus,it poses a significant danger to aquatic habitats,leading to a rise in temperature,and it produces eutrophication,which harms the aquatic ecosystem.The direct economic losses caused by water hyacinth in China annually are nearly 10 billion yuan.If we could extract some effective insecticidal active substances from water hyacinth plants.In that case,this will provide a new approach to the comprehensive management of water hyacinths and could provide new methods for pest management.Therefore,the insecticidal activity of some extracts（using different solvents）of the E.crassipes aquatic plant against the black cutworm A.ipsilon larvae was studied in the present research.And its safety assessment against albino rats as a mammalian model was also analyzed.The main findings are as follows:1)The insecticidal and biological evaluation of E.crassipes extracts on second instar larvae of A.ipsilonThe insecticidal activities of five extracts of water hyacinth,including petroleum ether（Pt.Et.）,acetone pure（AP）,ethanol pure（Et P）,acetone fraction（AcF）,and ethanol fraction（Et F）extracts,against the second larval instar of the black cutworm A.ipsilon was evaluated by the leaf-dipping technique.The results showed that the AcF and Et F extracts had the most mortality（93.33%for 1000 mg/L）and（66.67%for 500 mg/L）,respectively.Accordingly,the highest toxicological activity of tested extracts was recorded following the LC₅₀ values as 240,370,490,2000,and 3900 mg/L for AcF,Et F,AP,Et P,and Pt.Et.,respectively.The findings also revealed a proportional relationship between the concentration gradient of all tested extracts（Pt.Et.,AP,Et P,AcF,and Et F）and the mortality percentage of A.ipsilon larvae.Furthermore,there were some disturbances in various biological attributes（adult emergence,larval and pupal durations,and pupal weight.The percentage of total inhibition of adult emergence was proportional to the concentrations of all tested extracts compared to the control experiment.The plant extracts potency arrangement was as follows:AcF,Et F,AP,Et P,and Pt.Et.2）Biochemical assessment of the potential E.crassipes extracts against A.ipsilon larvaeThe effects of extracts from water hyacinth on the total body metabolites（total carbohydrates,total lipids,and total protein）of A.ipsilon larvae and the activity of some enzymes,such as phosphatases（acid（ACP）and alkaline（ALP）phosphatases）and transaminases(alanine aminotransferase（ALT）and aspartate aminotransferase（AST）were investigated by the quantitative spectrophotometric analysis.Besides some detoxifying/resistance enzymes（glutathione-S-transferase（GST）,andα-andβ-esterases）and acetylcholinesterase（ACh E）in the body homogenate samples of A.ipsilon larvae treated with the LC₅₀ concentrations of the most potent E.crassipes extracts（AcF（240 mg/L）and Et F（370mg/L）.The total carbohydrates and total protein reflected a significant increase（P<0.05）（25.73 and 2.43 mg/d L,respectively）with AcF extract.The total lipid content exhibited a remarkable decrease in treated samples（232.47 mg/d L）,compared with 267.60 mg/d L in control.At the same time,no significant changes were determined with the Et F extract.Moreover,the activity of ACP and ALP showed significant elevations with both extracts（23.57and 86.36μg/mg）for AcF and（29.27 and 84.09μg/mg）for Et F,respectively.ALT revealed a significant increase with AcF-treated samples（61.40 mg/g）and a non-significant reduction with Et F samples（40.93 mg/g）compared to the control.AST exhibited a significant rise with both extracts（194.30 and 168.57 mg/g）for AcF and Et F,respectively.Additionally,AcF extract reflected a substantial reduction inβ-esterase activity.At the same time,Et F increasedα-esterase and GST concentrations（11.02μg/min/g body weight and 29.96 mmole/min/mg protein,respectively）.The activity of ACh E revealed a significant reduction in samples treated with both extracts（AcF and Et F）,as 65.05 and 89.84μg/min/g body weight,respectively,compared with 271.16μg/min/g body weight for the control group.3）Chemical composition of different solvents extracts of E.crassipesThe predominant ingredients of extracts from water hyacinth plants were analyzed by GC-Mass.According to the GC/MS analysis data of E.crassipes extracts,the major active metabolites represented about 92.44%,94.57%,and 88.73%for Pt.Et.,Et P,and AP,respectively.While the major compounds of chemical composition of the acetone fraction extract were thirty-one compounds,including n-hexadecanoic acid（16.72%）,linolenic acid（11.13%）,hexadecanoic acid（9.35%）,hexahydrofarnesyl acetone（9.03%）,phytol（3.79%）,2-phenylundecane（2.83%）,2-phenyldodecane（2.75%）,hexa-hydro-farnesol（2.60%）,neophytadiene（2.39%）,2-phenyltridecane（2.24%）,12,15-octadecadiynoic acid,methyl ester（2.21%）,8-octadecenoic acid,methyl ester（2.16%）,6-phenyldodecane（2.08%）,and phthalic acid with total 93.72%components.The Ethanol fraction extract reported thirty-one compoundsthat were identified.The main constituents formed about 85.12%as hexadecanoic acid,ethyl ester（7.86%）,tridecane,2-phenyl-（6.30%）,2-phenyldodecane（6.22%）,hexadecanoic acid（5.51%）,6-phenyltridecane（4.99%）,3-phenyltridecane（4.47%）,5-phenyltridecane（3.66%）,4-phenyltridecane（3.56%）,dodecane,3-phenyl-（3.49%）,oleic acid（3.39%）,undecane,2-phenyl-（3.37%）,hexadecanoic acid,methyl ester（3.04%）,and dodecane,6-phenyl-（3.01%）.4)Safety assessment of acetone and ethanol fraction extracts of E.crassipes on albino ratThe safety assessment of the most potent extracts（AcF and Et F）was conducted using their LC₅₀values to investigate their exposure effects on body weight,hematological indices（CBC）,biochemical indices（liver and kidney functions,total cholesterol,and Triglycerides）,and histopathological studies on some vital organs of albino rats.The obtained data investigated non-significant changes in the body weight of all treated rats during the experimental time.There was a non-significant effect on the values of white blood cells,lymphocytes,monocytes,neutrophils,eosinophils,and segmented cells with AcF-treated samples compared to the control.Conversely,Et F-treated samples reflected a significant reduction only in the monocytes（612±159.80）×10³/μL)compared with the control（1006±255.60）×10³/μL.While reflected non-significant alternations in total red blood cell count（RBC）,hemoglobin,hematocrit,mean corpuscular cell volume（MCV）,mean cell hemoglobin（MCH）,MCH concentration（MCHC）,and platelets count.The biochemical results of ACP and ALP showed no significant alternations in their concentrations upon the treatment with both extracts.On the contrary,a significant elevation（p<0.05）was detected in ALT activity in the group treated with AcF at 36.73±1.44 IU/L compared with the control at 25.04±1.99 IU/L.Urea and creatinine exhibited non-significant alternations with the extracts.The total cholesterol content and triglyceride level had marked elevations with samples treated with Et F（100.67 and 134.67mg/d L,respectively）.On the other hand,AcF results revealed a significant depletion in total cholesterol content（67.33 mg/d L）while a non-significant change in triglyceride content compared to the control group.The histopathology of some vital organs was conducted to investigate the safety and the acute toxic effect of selected extracts on the albino rats’tissues.The heart from the AcF and Et F groups revealed normal histology compared to the control group.At the same time,lungs from the AcF and Et F groups showed slight inflammatory signs in the form of a mildly congested blood vessel with a thick muscle wall and slight peri-bronchial and interstitial tissue mononuclear cell infiltrations.The liver from all experimental groups reported normal hepatic parenchyma with organized hepatic cords,normal blood sinusoids,central vein,and portal tract.In conclusion,the present research suggests that the acetone and ethanol fraction extracts of E.crassipes exhibited potent toxicological activities on the black cutworm Agrotis ipsilon and are safe on the albino rats.The results provided a foundation for controlling water hyacinth and developing a new insecticide.