The effects of natural biopesticide from Mirabilis

Biological control provides a safer alternative to reduce the population of agricultural pest. 15 Mirabilis jalapa is one of biopesticides containing chemical substances that have a feeding deterrent 16 property against Spodoptera litura as folifagus insect pest. This study aimed to analyze the humoral 17 and cellular immunes responses of S. litura after exposure to biopesticide extracted from M. jalapa. 18 The measured indicator immune responses were activity of hemocyte, lectin, phenoloxidase (PO), 19 and phagocytic activity. The results showed that the average total hemocyte was different 20 significantly depending on the treatment. Exposure to 0.1% and 0.2% (w/v) of M. jalapa extract 21 increased the total number of hemocytes as much as 38.08% and 64.15%, respectively. Lectin was 22 quickly formed at 0.1% and 0.2% (w/v) concentrations. The amount of PO enzymes was significantly 23 different at sublethal concentrations compared with control samples (P < 0.05). The highest increase 24 in PO activity occurred at 2 h post-treatment and at M. jalapa extract concentrations of 0.2% (592.33 25 IU/mg) and 0.1% (521.33 IU/mg), whereas the highest concentration of the extract (0.8% w/v) caused 26 a decrease in lectin and PO activities. In terms of phagocytic activity, the proportion of phagocytosis 27 cells were 47.62% in control group, and decreased significanlty in both concentrations exposure. 28


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can lead to up to 100% defoliation in crops [12]. The resistance of Spodoptera to various chemical 51 compounds needs to be monitored because these pests are quickly spreading across the Asian and 52 South Pacific regions [13]. Therefore, it is extremely urgent to control these pest populations using 53 alternative methods, particularly plant-based biopesticides. To control the pest population, its 54 immune system must be clearly understood. In this study, we evaluated the potential of a 55 biopesticide extracted from M. jalapa to control the insect pest S. litura by weakening its immune 56 system.

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The main role of M. jalapa as a biopesticide is to weaken the Spodoptera immune system. The 58 immune defense mechanism acts as a barrier to infections when exposed to foreign agents; it 59 biochemically responds when attacked by a foreign agent. Therefore, the state of the immune system 60 can be used as an indicator of the potential for pest mortality. activity. Hemocyte acts as the main subject of cellular immunity to recognize, tolerate and eliminate 65 the presence of foreign in the body. The insect's immune system conditions will be activated when 66 exposed by foreign substances. The hemocyte is working to eleiminate foreign by the phagocytic.

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However, each type of hemocyte has a different function in insect defense. The whole function of the 68 cells will work together doing the phagocytosis mechanism. Thus, activity of hemocyte and 69 phagocytic are main role in cellular mechanisme as active (amoboid) cells. Therefore, phagocytosis 70 becomes one of the parameters in the cellular defense mechanism. A test of the effectiveness of 71 phagocytosis from insect larvae to determine changes in the immune response that occur in the body 72 of the insect needs to be done when exposed by a toxic substance.

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The humoral response is a very crucial part of the insect immune system because it plays a role 74 in activating protective enzymes and stimulating the ability to recognize pathogenic invaders.

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Therefore, this humoral mechanism acts to stimulate the functioning of the immune system and 76 therefore, the humoral response is one of the parameters of the immune system which can kill the 77 infecting pathogen.

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Previous research on pest control using biological agents has been widely conducted, including

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Unfortunately, most biopesticides are considered to be harmful to the ecosystem if applied over a 85 relatively long time period. Therefore, when using a natural compound as a biopesticide, the 86 concentration are very important parameters when trying to deter pest resistance. However, to the 87 best our knowledge, there have been no previous studies examining the use of natural compounds 88 in M. jalapa to weaken the pest immune system to decrease immunity cause mortality. We believe 89 that this approach can potentially provide an ecofriendly way to control pest outbreaks because the 90 use of sublethal pesticide concentrations is feasible for farmers.

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The aim of this research was to analyze the effect of M. jalapa leaf extract on the immune

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HA assay was performed using blood from vertebrate animals containing anticoagulants. The

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blood sample was washed three times using TBS at pH 7, and its concentration was reduced to 2%

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The hemocytes changes aims to maintain physiological balance and endurance thier body. Therefore,

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The humoral immune response in insects plays a major role in the immune system by activating

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Our results showed that lectin was present in S. litura when it was exposed to M. jalapa

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The formation rate of the titer was influenced by the number of hemocytes produced by cells,

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The activation of PO is the main enzymatic reaction important to the humoral response sequence.

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The next stage of phagocytosis is the endocytosis of B. cereus bacteria into the plasmatocyte cells.

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The plasmatocyte cytoplasm contains many lysosomes that are responsible for catalyzing the alien   pests in a given field is required for one to establish the amount of biopesticide needed to be applied.

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The magnitude of the impairment of the immune system observed in this study was enough to kill 435 the target pests. Therefore, it is has great potential for controlling insect pest populations.