2.1 Choice of plant material –
Thirteen plants were selected and kept in consideration for the development of herbal formulation. These plants have medicinal properties. Some of these plants were tested by researchers for antifungal, pesticidal, and antimicrobial activity (Manoj Kumar et al., 2021; Narsimharaju Kalidindi et al., 2015; Bachchu M A et al., 2017; Hosseinabadi 2021; Kotkar H M et al., 2001). The medicinal plants selected for the current study were - Gloriosa superba, Vitex negundo, Ipomoea carnea, Annona squamosa, Calotropis gigantea, Pongamia pinnata, Jatropha curcas, Tinospora cordifolia, Terminalia billirica, Datura stramonium, Azadirachta indica, Capsicum annum, Allium sativum.
Above mentioned Fresh 13 medicinal plants were collected from regions of North Maharashtra. The plants were selected based on available literature, abundant availability, medicinal and insecticidal, and various pharmacological properties. All the collected plant specimens are identified from the Department of Botany, Dr. P.R. Ghogrey Science College, Dhule.
2.2 Formulation of the product (Preparation of plant extract)
- Liquid formulation
- Dry formulation
Liquid formulation includes water-based, oil-based, polymer-based, or combinations. Here I choose a water-based formulation for foliar application.
Dry formulations can be dust powders or granules. Dust form was used to check the efficiency of herbal formulation as a stored grain protectant. I chose the method to make dry formulation was air drying (Ukoroije 2020).
- Selection of plant material
- Choice of solvent
- Extraction methods
The basic principle of work is to grind the plant materials (dry or wet) finer, which increases the surface area for extraction to increase the rate of extraction. Fresh plant parts (dried or wet) are ground in a blender to fine particles, put in a certain quantity of solvent (Water), and shaken vigorously.
All collected leaves were thoroughly washed in distilled water thrice to remove dirt and were allowed to drain on a wire mesh. 20 grams from each of the dried, powdered plant samples were weighed. This material was soaked in a 2.6-liter mixture of cow urine and distilled water. Allow this mixture to mix properly and keep for 30 days. Shake it regularly twice a day.
After 30 days, the extract was filtered with the help of muslin cloth or Whatman filter paper no.1. Cold maceration process was selected for the extraction of phytoconstituents. (Maceration (for fluid extract) is an extraction procedure in which powdered plant material (leaves or stem bark or root bark), is placed inside a container; the solvent (distilled water) is poured on top to cover the material completely. The container is then closed and kept for at least three days (or until the soluble matter is dissolved) The content is agitating frequently or periodically, and if it is placed inside a bottle, it should be shaken regularly to ensure complete extraction. At the end, the micelle is separated from the marc by filtration or decantation. This method is convenient and suitable for thermolabile plant material.)
2.3 Larvicidal activity
2.3.1 Collection and rearing of Tribolium castaneum – The Adult Tribolium castaneum was collected from naturally infected wheat flour from the local market of Dhule. Beetles were reared in wheat flour and taken in glass beakers as food medium. The beakers were kept in an incubator at 300 Celsius temperatures without the control of light and humidity. About 50 adults in each beaker were introduced with 50 gm of wheat flour. The cultures were checked regularly. The beakers were covered with muslin cloth and tightly fixed with the help of rubber bands to avoid the escape of the beetles (Maheswari M et al., 2017).
The larvicidal effect of formulated biopesticide carried out against red flour beetles (Tribololium castaneum).
2.3.2 Bioassay test (mortality test) – The residual film method (Busvine 1971) was used to larvicidal test of different plant extracts against 6th instar larvae of Tribolium castaneum (Herbst). 1 ml plant extract was dropped separately on Petri dishes (60mm) with the help of a pipette, covering uniformly the whole area of the Petri dish internally. The petri dishes were kept open for some time to evaporate the solvents. Then 10 larvae of the 6th instar were released in the Petri dish. For the control treatment methanol solvent was used. 3 replications were made for extract treatment (Bachchu et al., 2017). The Petri dishes were then kept without food in the laboratory and larval mortality was recorded at 12, 24, 36,48,60, and 72 hours after treatments (HAT). The corrected percentage of mortality was calculated using Abbott’s formula (Abbott, 1987; Puntener,1981).
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m0KgwmG5iAPXz4sFVRJsXTV+kmZe5/gdqZPwpap/atMoN5f+5bqW2tVeF6Ol62UsARF12rdhWO35544ommxr0sAUe5Bw8e7C3A1hVJ6zDTTw3XqvpgPH0m4BAgDDZZq3D4M/jEfGL4G8wfq1ev9uZ4+CUwML7zzjtu2LBh/lUMBgcOHGhszIJJGent3bvXh4MPmQ8nEqiIVatWNfzuMOFjEmE31OCdsMJRoYgXgrVv3z6/8oUFgf5+mw7yEKaLf1sfTpYJEWWEReLSpUs+ilDAy/q9mQ/Liivm8Df63vEOTFeh2d42Tppy7W+3b9/2Ps7PPvvMb1IaOnSo74hgzPIgzcWLFzfqFB1kyZIljVk2yof2QIaYha9Zs6axlyIrXbs56tVXX/XpIg6kfezYMTdixAhfX9xvgb0Dc+bM6dLfIBhbtmzx/PHesmXLGmFSWcXy18oJDtpDJ/etmICrb8WHfQgKNvxx/Iv5ZdFn9+zZ02izGMusaRlxfPXVV96qB3cbxz7bL8Af/SwcLxG+qD8XjdEcpzHhTHWb4B2M02jrKPvu3bv9ggX5C03qqYKZJeDWJB/TAHC6ceOG1yLsf4qVAXEzXGp+UsP1qYDTDBHzZWBljM1iEMWYgGMwXb58uTt+/LgXS3T8adOm+XKjUiHiVgwAFoKBlf+2bdv8gB+rNDQKVAjSxrNgwQIvyvA5Y5DnE77LjRAQALvapijb36wJiMLFeJH+K6+80k08+Pc8H3hP/N6WFQUlxm/IkCENgYWQcbIUa3CcoDE+DAQQUpi/MKOFHwncwAD/Y2cHb8RLTnaWC1/UpEmT/KQM8eG/MeGwZvqidNGZpkyZ4q5du+bzgskb0oPpzZbP1jnEHQMEfGB4ONBx/0EZVmH+UjtrSjgKeKf2rZiAq291r3m0x9OnT/uxCW2X44wVEE72w/4EMUZ/w3jx22+/eUsofkNfGjdunE/sjTfecI8//rgfC7PGvKL+nDJGNyvgeM9OLMIVckpfsWGyBDzG1fLAf2N8Sw1XNl9F4ftUwJE5CDWEwIqBNQHHwNB8gZmXXSXFBi0OljGxiVUaVkgY4Gku4YASmsBj78bEOvYbyo18Xbx4scuOb5qpbKeJVaBdCduNFNbvjbg4+bCz6bwGERMW/obVM31NqcflYvGRZ9akjZMr5pMrVtQfHgw0dnVMvnYVmyWQqLNw136sfOGOZ7Y3y4AmMzuzTmXVGwLeqX0ry4SuvvX/PZft0Z7ioZhxPGAfDRceND3bja/oFxR1psJwtl/Fxrys/lxmjC7jNgnHL07y0Q+x6Gr2+KsEvGiqYP7OVQJ+iokuRB2zy6yZZZ7vC7NGrJTY8PIGy7yNCzTbYUWN+Fot4FhVYAe5FZSe+K4hOlevXm2Y+tEBMcuGORVWDBw7KzoelifgtiNnTWrCJpAn4KGpiTyymlHInyZrmrxTBTxMNyWPbG9ZectrazFWvSXgndi3WiHgde9bsUlr2Daz9lmQjd17lLcYSRXwsF+VGaObFXBa4DD5gJWhJ6twCXiTAh6uFLAq/e677xq+jNgKPGtVy5UtfJvNCjh94JhAYIc6jn/Fznb3dAWOvHKWS+uAnbiUwOlNSYjLnmFE/tip2Gljx81sOn0p4KmTAvrAke9Fixb5SQtcLO0U8JQBM9b28FtPBBx1GD5F/nI7Oe7EvtUKAa9730ppj2UWLu0Q8DJjdDMCTnchxji4vaw/PNy7kjKWZgk4J8FZvm1aLlLDpeSlTJg+N6Ejs6xsmH8gRtiIQN9qnoCHJnQOopgE0CdapiFzY0RvmNCRV5qZMHN8+eWXu0xcylQiymiZxTqEFfSsuPtSwPMmGeD0999/u//85z/eb94TE3pPVuAxNww6Ljfz5a3mrQUhdQVuN3KyzsKNNDGrh53IdVrfapWA17lvsU5jF2GxPbKNhSZ0tBeMBXa12k4BTxmjywo4x2lshuW+JIwfGBtgLY2VuWg8zRJw247sLnSOV3QFpoYrykfZv1dCwHkuFpnHYGh3Eub5wGM7+2BCt7sxywg4G5JtdO3ygbOiuArHLvpz58512SiXUpnYzIKNJ+GNdp22AkdZUXd4Qg5ghDbBtmA7aFkfeDMCzs4anjtFu/34448blzm0WsBT6r9IwDutb7VKwO0qvG59K6s94ndYHzHJy9rEyHfthLIdAk5BSxmjywg4xXvp0qXdTsFw7wrqu8hdGPabsufA6SKwk/rYOfBYuGb6ddY7vS7g3GwRHkPKMrnQlxPexMYjWpgB2Z3B9PtiM4M9ahCbrWKDFCrg888/dydOnPAb4uCX5tGkBw8e+CMW8K/AV02hZCcIN05QjO0EgI0K+cQuZ05SWCHkMWbMmNLn0XmLUOwmOesDh8gUXZFqWVmB5CYy+xsbZdGxD9YRj3ng+AqsI7gvHj648JY4Djo8EogznWfPnvWmckxQmC7awsKFC92PP/7o/45BC+4BmNLs8T2bLlbw2G1rVx5ZZQ53v1sxQJ0jjoEDB7ojR440dgEjTCqrGJe83fypHb4OfcvWsV3xqG91bQXkwWOu9+/f923RbuRie+R4hPaBtovfuUiymzHtuxx3rdCH4yXiYJ+N9eeUMRqlyqrzsN3zlApW2bENaywL3sMY/sknnyRdZJU1IQrHaC4ueeIpvImN/a8oXGp/TgnXqwLOFYrNGH163DUd7v4OCxEex+K5XA76EHPuRAx9iKHgIC10BKz87FlgnN1GfDDVPv/8874xQGAhODx2YfOFPMFfbs93My36avC3rHvLUeEwoeedqw45xPzeNow9z56yCz1khTKFdw+jTBRMppV1EQ/+jo4xffp0785AZ8dZzfCu4tCna8+NIt/gSi6WJf6G8/aYZGGPgmWbmm6sfeSVz56pRZuwJrwycYX5K1PvWZ26Dn2LR0bD8SEsm/rW/xGy30SAMGPcshNBWoi4ETe804Ir33Asi/V7rurteIkxrag/27sTYmN07P7y2D6PonCxsqBcRYuMWL+JXbpl758I74Cw/FLDpYhzSpheFfCUDPW3MKlHx/obF5VXBHpKQH2rpwT1ftUJSMD7uIYws4Q5tpmdk32cdSUvApUmoL5V6epR5lpAoFcFPHYspgVlUBQiIAIiIAIiUHkCRcdAyxagVwW8bObqGp4bUNr1idS6clO5RKCIgPpWESH9vU4EJOB1qk2VRQREQAREoN8QkID3m6pWQUVABERABOpEQAJep9pUWURABERABPoNAQl4v6lqFVQEREAERKBOBCTgdapNlUUEREAERKDfEJCA95uq7p2C4oa4o0ePuj179vib0nC1KR98bQ3/sx/b6J1c9TwVfizB3rPf81jrEUNendejhCqFCFSTQL8TcA42+D42v1gWVk14BSHvWw8/Fp8aLrXqcY3o/v37/Z3GeG7fvl34gXoIIu5z59Pqc4apeWc4CB2uoEUZwisJcZcyGLZTwO0Vsri6Mbxz35anzPfXec+xBLx7i8ir87LtR+FFQATSCfQrAefXeiDeuBA/S+xwPy6EACKEW9Lwbzz4UIp9UsOlV4fz6Q4ePNi/UnSPLz8kkBK2TB4YFhMKfA/drqJT4sn7FnDsfXz0IPZBlpS0wjCIix91gCVg7ty5foITfiyEomM/mNFMep32DtoX7hwv+ixp2XKVrfOy8Su8CIhAdwL9SsBZ/LxPjFIU7eqRn8azgpoarplGx6/+4N28z+IhHCYiEKjYBfzNpB1OUFauXNlWAefnLrOsIWXLEH5OFrf/hd8k5kdRwq+hlU2rE8PzG+MS8E6sPeVZBLoSkIAHLYLiGQoKTNV4bty44f8/NVwzDQ4TjOeee85/6SdrFY6v72zfvt3du3fPfwWt1QLOT/w1E2+Z1Rg/W9gq07/9DjrYh//Gb7ita9asWaUnJs3UZZXe4WcTiyw7zeS5TJ03E7/eEQER6E5AAm6Y0Hwd+0QmV+1Y7cKsDjN3Ubhmv/GMtLA6HD58uM9dbBWOMFgd8zOmMaG1n7/kpwTxTW768mFFwPfOMQnAoM5Pf06aNKnhhycexm8/l4e/8dvp+F43n9hgbtOiD5zfK7bNEkxjn2VFGPvpv6yJBVbg9ju9oYCX8XszX2gXMMfj86EzZsxomJ9tmVBfq1at8t9dT/l8K+KmORsb/tCu8N62bdv8t89t2ogXacHaAt5Lly7t8olVmzfww8QPDO03mvldessadf7MM8+4AwcO+PjRzmbPnu3fPXnyZCMfRe0IcUrAJS8i0PsEJOCGOb8pmyfMEA48+GZuUbiyvmNmBUIFkeO9zgcPHuzyrXCKKMJQ1EJBw8oWogB/MCYSXFFDAODLh3jw2+b4DcKEB2Jy+vRpL1gQAhsvzd0QeAz4cC3gv2GtsBaLcDC3aYXMQneG3YQW+346BPnKlStuxIgR0d6S5wNv1u+Ncp45c8b70+23qC0/cBg3bpzP0xtvvOG/MU9rTVa3xgQG/8M37PEsWLCgy+Y/fg4TcTMMJwlkY/OGu/UxCZgyZYq7du2azy8mM9jIh4ft267A8dvixYvd5cuXfdmGDh3qJ4acSBS1I5ZNAt77g7dSFAEJeIUFnL53DMpWDKwJOCbg9M+HQhebEEAQw/izVlTciY1d5vz8KQduawLPGsyRVpGAI21OFMaMGdNlxzrdBnkbz+wEAOWChQIiFPq9IeZvvfWWFy67Us0aEmLih7AoEydFfJec89wCrCN70oCrZK5+EQ/Cxdw5mJxxcx7zFk54wmN7WWWw1iVrNSrTjiTgEhMR6H0CEvAKCziyFoouV70U9JiAZ/mV6QO1IhoT1SwBt80TYgDTK0zGeFop4IiPFgNryu2J7xorc6xMaRWBuGFli9UpTxnkHW/LE/BwUhKb1IRdOyVNrOLDSYxlw5V0njAjPMtVJODhhKNMO5KA9/7grRRFQAJu2gBXvHmmcayY7t+/71etReFoYrXNLPZObHDnoBuuwiFiEydObJjUYwKet8s+FOyyAk7zPfK8aNEid/XqVW9qb7WAI367cTCcuJTpuljZnj9/vnFUjUJG9wAnC7HjZkynLwQ8q27CvLRLwMu0Iwl4mRapsCLQGgIS8IAjRCP06SIIVkPWr5kSjpfG2CTgY8TKJu+hD5xhOJDiOBQ2Utnz6HkCHvMVQxTgK6UZuoyA8zaydpvQWW6acFFupG0nLqnNH3WASY9dXYdiEwp6LO52CDg2i8Uu60G50UZQN7iMJjShMy88HtduAU9pRxLw1BapcCLQOgIS8IBl7Hw3Tc92M1lquGaqKhRwDtCxm8XyfOB2AxPywXLYjWllBJyDtB3Q2+EDt8w4UcJvzZwVBx+Yya1vtworcK76wzpCu8LlOdiJnrWJEe8uX768cTqhXQLONp7SjiTgzfR0vSMCPSPQ7wScO3vtJqAQIXYG48GqDeZyrFjxhDexpYYrU0XcyBSacyFEFy9e7HKkzJYlPNvLI1pcpWElinLgd+5KpnkeEwNs5rIix3xg8MbKFw9YYGcz4lm4cKH78ccf3dmzZ/2xI0xuhgwZ4n3M9J3aC1SsK+DChQuNo2zwTW/YsMFftnLr1i2HY26xfDRzdhlxY2e4PZbFuqAPHLu7cXSK9Z1VVxQza73gsUO4U2yZWP68M/R4F3XKTXQ49//HH3/4nf3WfYINcniOHz/u2UKsp02b1uWKWNZVmLexY8f6SQ+PITK/PPoH9wd4Mx92vwE5pLQjhI3VeZl2r7AiIALlCfQrAecqwWKKCUN4nzZEbMmSJd3uJU8Nl1otWA3bx/rLMXBDMHmDVqwseJe+6PCedp4D5/tctdn0wiNjPBPM3c3cyQ3BRnz4WMmDBw/c1KlTHcPYs9qIG2XAsSQcu7MP8wmrAM+fZ91bDtdFOMEoYhr6vcPw9jx70S70kHVWmcAvLGfexCOl/UDQwZb344fn7mPtIJYP1i0nTCwzr+0lnzC/Re0I78XqvJ333RfVvf4uAv2FQL8S8P5SqXUqJ4T4xIkTDatBncqmsoiACIhATwhIwHtCT15Kf94AAAGvSURBVO+2nQBWd1gVZl3c0vYMKAEREAERqCgBCXhFK6a/Zsv65WE651nt/spD5RYBERCBLAIScLWNShHg5i5s4rN3mlcqk8qMCIiACFSAgAS8ApWgLIiACIiACIhAWQIS8LLEFF4EREAEREAEKkBAAl6BSlAWREAEREAERKAsAQl4WWIKLwIiIAIiIAIVICABr0AlKAsiIAIiIAIiUJaABLwsMYUXAREQAREQgQoQkIBXoBKUBREQAREQAREoS0ACXpaYwouACIiACIhABQhIwCtQCcqCCIiACIiACJQlIAEvS0zhRUAEREAERKACBCTgFagEZUEEREAEREAEyhKQgJclpvAiIAIiIAIiUAECEvAKVIKyIAIiIAIiIAJlCUjAyxJTeBEQAREQARGoAAEJeAUqQVkQAREQAREQgbIEJOBliSm8CIiACIiACFSAgAS8ApWgLIiACIiACIhAWQIS8LLEFF4EREAEREAEKkBAAl6BSlAWREAEREAERKAsAQl4WWIKLwIiIAIiIAIVICABr0AlKAsiIAIiIAIiUJaABLwsMYUXAREQAREQgQoQ+C86VHqOtU6PKwAAAABJRU5ErkJggg==)
2.4 Insecticidal activity
2.4.1 Collection of insects
The mealybugs were collected into the glass petri dishes from the unsprayed infested plant of Hibiscus rosasinensis of the family Malvaceae located in the Deopur area, Dhule. They were maintained in the laboratory of Biotechnology under conditions at 250-270C, and relative humidity 60 - 70%. Healthy and active adult mealybugs were used in all toxicity bioassays.
2.4.2 Toxicity bioassay
Four large-sized glass petri dishes were taken for the experiment. Each Petri dish contains leaves and 20 adult mealybugs. The collected leaves were washed thoroughly twice with distilled water and completely air-dried before use for the application. A slightly moistened filter paper was also placed in each petri dish to keep the leaf turgid during the bioassay period (Samman Fatima et.al., 2016). The prepared solutions of four concentrations of botanical extract (4 %, 6%, 8%, and 10%) were sprayed over the mealybugs, and the container was covered with a muslin cloth to avoid the escape of mealybugs from the petri dish (Devi et al., 2018). The leaves which are sprayed with distilled water are considered as control. The mealybug mortality was observed after 24 and 48 hours following initial application. The adult mealybugs who did not respond after pin probing or not moving when touched with a fine brush were considered dead. The mealybug mortality records were obtained in percentage value. Two replications for each treatment were carried out (20 insects per treatment) (Govindaiah et al., 2006; Fatima et al., 2016).
Percentage corrected mortality was calculated by the formula (Puntener, 1981):
![](data:image/png;base64,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)