| Neem oil is meliaceae melia plant neem tree (Azadirachta indica a. Juss) seed extract, as one of India’s traditional medicine, has the parasitic resistance, anti bacteria and fungi, antipyretic analgesic, immune regulation, fight ulcer, and promote wound healing and other pharmacological activities. Octadecanoic acid-3,4-tetrahydrofuran diester isolated from neem oil had acaricidal activity of new material, about its research focused on the substance of separation, purification, structure identification and acaricidal activity in vitro, the mechanism of the acaricidal effect research is not reported. In order to develop new types of botanical acaricide and make full use of the rich neem resources in our country. This study discusses the acaricidal activity of octadecanoic acid-3,4-tetrahydrofuran diester against Sarcoptes scabiei var. Cuniculi in vitro, on this basis, through optical microscope and transmission electron microscopy (tem) method, the observed after drug mite bug body poisoning symptoms and ultrastructural changes; and the insect body related enzyme activity were determined. The main results were as follows:1. Study on the acaricidal activity of octadecanoic acid-3,4-tetrahydrofuran diester and four kinds of traditional Chinese medicine extraction against S. scabiei var. cuniculi in vitro:Measured octadecanoic acid-3,4-tetrahydrofuran diester, matrine, tuberostemonine, daphnetin and artemisinin as experimental drugs, were used to kill way to determine its acaricidal activity of S.scabiei var.cuniculi larvae in vitro. The results indicated that drug concentration of10.00mg/mL, octadecanoic acid-3,4-tetrahydrofuran diester, matrine, tuberostemonine, daphnetin and artemisinin onset time for were50.00min,45.00min,127.50min,165.00min and263.50min, all the time of death for1485.00min,1455.00min,1890.00min,2130.00min and2280.00min, respectively. The median lethal concentration of the matrine (at the concentration of25.00mg/mL) was2.859mg/mL at24h post-treatment. At the concentrations of25.00mg/mL,12.50mg/mL and6.25mg/mL, the median lethal time of the petroleum ether extract was7.544h,9.848h and14.549h, respectively. Conclusions:The Five Herbal monomers have some acaricidal activity in vitro. Which most acaricidal activity of octadecanoic acid-3,4-tetrahydrofuran diester and matrine. followed by tuberostemonine and daphnetin, and acaricidal activity of atremisinin weakest.2. The influence of the morphology and ultrastructure of tissue of octadecanoic acid-3,4-tetrahydrofuran diester against S. scabiei var. cuniculi in vitro:With a concentration of20.00mg/mL in the octadecanoic acid-3,4-tetrahydrofuran diester solution after treatment of rabbit scabies mite,in different time intervals were observed under optical microscope mites symptoms of poisoning, and the ultrastructure was observed under transmission electron microscope. The results show that:under light microscopy, mite within24h after drug treatment, first kept movement, movement disorders, spin around, have front feet twitching excited expression;Diffusion is mite bug to escape again, slow motion, feces, spawning sharply;Last mite body motionless, the body begins to shrink to death; Under electron microscopy, mite insect body wall leather layer is smooth;Dermis cell membrane dissolved, mitochondrial morphological abnormalities, spinal disorders fall off; Body cavity rough endoplasmic reticulum cells increased, the mitochondria vacuoles, cell nuclear membrane damage, nuclear chromatin dissolving of such lesions.3. The influence of enzyme activity in the protective enzyme, detoxifying enzymes and neurotransmitter affect enzyme of octadecanoic acid-3,4-tetrahydrofuran diester against S. scabiei var. cuniculi:With concentration of20.00mg/mL,10.00mg/mL and5.00mg/mL of octadecanoic acid-3,4-tetrahydrofuran diester and liquid paraffin processing rabbit scabies mite, respectively in8h,16h and24h proenzyme extract preparation of worm body, by ultraviolet visible light spectrophotometer, respectively superoxide dismutase (SOD), peroxidase (POD), glutathione-S-transferase (GSTs), monoamine oxidase (MAO) and Ca2+-ATP enzyme activity.The results show that within24h,different concentrations of octadecanoic acid-3,4-tetrahydrofuran diester solution treatment after rabbit scabies mites, the insect body of SOD and POD activity were significantly inhibited;Concentrations of20g/L, the SOD and POD activity inhibition rate up to93.92%and80.30%, the concentration of5g/L, the SOD and POD activity inhibition rate as low as47.68%and16.67%, showed that the higher drug concentration, enzyme activity is suppressed.Different concentrations of octadecanoic acid-3,4-tetrahydrofuran diester solution treatment after rabbit scabies mites, GST activity was higher than the control group in vivo, Concentrations of20g/L,the GST activity inhibition rate up to83.68%, the concentration of5g/L, the GST activity inhibition rate as low as10.38%, drug group of glutathione-S-transferase has certain activation mites in the body. The worm in the drug, the GST activity is activated.Different concentrations of octadecanoic acid-3,4-tetrahydrofuran diester solution after rabbit scabies mites, the worm in the body of MAO and Ca2+-ATPase activity was significantly inhibition; Concentrations of20g/L, MAO and Ca2+-ATPase activity inhibition rate up to77.34%and46.21%, the concentration of5g/L, the MAO and Ca2+-ATPase activity inhibition rate was7.72%and2.79%, and the lowest drug concentrations at high concentrations, the enzyme activity inhibition; Drug concentration is lower, the enzyme activity inhibition, over time, the enzyme activity inhibition rate increased. |
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