Bioinsecticide Production Research Articles

Evaluation f the Insecticidal Activity of Nerium Oleander and Artemisia Campestris Powder and Extract on Wheat Insect Pests

Objective: The study aimed to evaluate the insecticidal potential of powders and methanolic extracts of two plants of the Lamiaceae and Apocynaceae families spontaneously growing in the region of Tiaret. Method: Insecticide tests were carried out in the laboratory using the Direct Contact Method. Results and Discussion: The obtained results showed that Nerium oleander and Artemisia campestris powders have remarkable insecticidal properties. N oleander powders induced 100% mortality in adults and up to 80% mortality in Tribolium castaneum larvae at a dose of 0.5g, after 7 days of exposure. Adults of Tribolium confusum showed some resistance to the powders of both plants, with an optimum mortality rate equal to 50% for adults. Larvae of both species were found to be sensitive to N. oleander powder with a mortality rate ranging from80to 90%. As for the powder of A. campestris, adults of T. castaneum were more sensitive than those of T.confusum, with adults’mortality rate equal to 100%. The methanolic extracts of both plants showed a very remarkable efficacy against the larvae of both species as well as the adults of T. castaneum with a mortality rate equal to 100%, after 7 days of treatment at different concentrations. Research Implications: Algeria has an abundance and diversity of flora that could provide new sources of compounds of plant origin with phytopharmaceutical properties, these results allow us to affirm that powders and methanolic extracts, of the studied plants, can be used for industrial application in the production of bio-insecticides. Originality/Value: This work could provide alternative or complementary solutions to the use of synthetic organic pesticides to protect stored beans. The development of bio-insecticides derived from plant extracts and the selection of varieties resistant to this pest are part of sustainable agriculture and development.

Long-term push–pull cropping system shifts soil and maize-root microbiome diversity paving way to resilient farming system

BackgroundThe soil biota consists of a complex assembly of microbial communities and other organisms that vary significantly across farming systems, impacting soil health and plant productivity. Despite its importance, there has been limited exploration of how different cropping systems influence soil and plant root microbiomes. In this study, we investigated soil physicochemical properties, along with soil and maize-root microbiomes, in an agroecological cereal-legume companion cropping system known as push–pull technology (PPT). This system has been used in agriculture for over two decades for insect-pest management, soil health improvement, and weed control in sub-Saharan Africa. We compared the results with those obtained from maize-monoculture (Mono) cropping system.ResultsThe PPT cropping system changed the composition and diversity of soil and maize-root microbial communities, and led to notable improvements in soil physicochemical characteristics compared to that of the Mono cropping system. Distinct bacterial and fungal genera played a crucial role in influencing the variation in microbial diversity within these cropping systems. The relative abundance of fungal genera Trichoderma, Mortierella, and Bionectria and bacterial genera Streptomyces, RB41, and Nitrospira were more enriched in PPT. These microbial communities are associated with essential ecosystem services such as plant protection, decomposition, carbon utilization, bioinsecticides production, nitrogen fixation, nematode suppression, phytohormone production, and bioremediation. Conversely, pathogenic associated bacterial genus including Bryobacter were more enriched in Mono-root. Additionally, the Mono system exhibited a high relative abundance of fungal genera such as Gibberella, Neocosmospora, and Aspergillus, which are linked to plant diseases and food contamination. Significant differences were observed in the relative abundance of the inferred metabiome functional protein pathways including syringate degradation, L-methionine biosynthesis I, and inosine 5'-phosphate degradation.ConclusionPush–pull cropping system positively influences soil and maize-root microbiomes and enhances soil physicochemical properties. This highlights its potential for agricultural and environmental sustainability. These findings contribute to our understanding of the diverse ecosystem services offered by this cropping system where it is practiced regarding the system's resilience and functional redundancy. Future research should focus on whether PPT affects the soil and maize-root microbial communities through the release of plant metabolites from the intercrop root exudates or through the alteration of the soil's nutritional status, which affects microbial enzymatic activities.

Effective medium for Photorhabdus luminescens bioinsecticide production and exploration of optimal mixture with Bacillus thuringiensis subsp. aizawai against Plutella xylostella

BackgroundPlutella xylostella is a major pest of cruciferous vegetables worldwide. We evaluated a biopesticide using different culture treatments of Photorhabdus luminescens (ATCC 29,999) and Bacillus thuringiensis subsp. aizawai and examined their insecticidal activity against Pl. xylostella. MethodsL8 and L9 Taguchi orthogonal designs were adopted to evaluate the optimal culture medium for enhanced insecticidal activity. Feeding bioassays were conducted to determine the potential synergistic effects of combining B. thuringiensis and Ph. luminescens, and the most effective synergistic combination was identified by analyzing the time–mortality response against Pl. xylostella. Significant FindingsAnalysis of the pH value indicates that Ph. luminescens has enhanced tolerance to acidic conditions and can alkalize the medium. Among the various medium combinations, Ph. luminescens optimized using A3B1C3N2—d10 × concentration, 24-h cultivation time, 1.25 % molasses as the carbon source, and a nitrogen source consisting of yeast extract powder, yeast powder, and amino acids (0.2:0.3:0.5 %)—resulted in the highest insect mortality. Synergistic interactions between B. thuringiensis and Ph. luminescens were observed in the 1:1 mixture, likely due to immune system suppression. These results highlight the potential of combining Ph. luminescens and B. thuringiensis to prepare a potent bioinsecticide against Pl. xylostella and other lepidopteran pests, thereby decreasing the reliance on chemical pesticides.

Insecticidal activity of Bacillus thuringiensis strains isolated from tropical greenhouses towards Cydia pomonella and Spodoptera exigua larvae

Three Bacillus thuringiensis strains were isolated from a specific habitat of tropical greenhouses located in the Botanical Garden of Adam Mickiewicz University in Poznań, Poland. The BG11, BG12 and BG15 strains harbour genes encoding a specific set of insecticidal proteins (cry1Ba, cry1Ia, cry2Ab, vip3Aa)—entirely different from those found in commercial isolates, currently used as bioinsecticides. Despite high genetic similarity of the new strains, each of them produces unique Cry1Ba toxin as a main component of the parasporal crystals. Moreover, the tested entomopathogens contain genetic determinants encoding two types of chitinolytic enzymes ChiA and ChiB. The tested strains display insecticidal activity against two distinct, economically important pest insects, Cydia pomonella L. (Lepidoptera: Tortricidae) and Spodoptera exigua Hübner (Lepidoptera: Noctuidae). However, BG12 and BG15 strains are significantly more active than BG11 towards both pests. The BG12 and BG15 strains can be considered as candidates for the production of new lepidopteran-active bioinsecticides with high potential to augment the existing biocontrol strategies.

BUDIDAYA PADI SAWAH ORGANIK UNTUK PEMULIHAN KESEHATAN TANAH DAN PENGURANGAN RESIDU PESTISIDA PADA PRODUK HASIL PERTANIAN

The tendency of more intensive use of chemicals in agricultural cultivation activities causes an imbalance of other nutrients and decreases the content of soil organic matter. This causes degradation of soil fertility and residues of active pesticides that threaten the sustainability of farming. The community partnership program (PKM)aimed to provide solutions to problems faced by partners in both the production and management fields by providing counseling, demonstrations and assistance. The method activities is participatory rural appraisal (PRA) in applying appropriate technology and empowerment, with partners in these activities being members of the "Batu Sari" farmer group in Astina Village, Torue District, Parigi Moutong Regency. The results of the implementation of community partnerships show an increase in knowledge and skills and attitudes of farmers. The production of organic fertilizers, vegetable pesticides and bioinsecticides carried out by the activity participants went well and could be used as expected. The system of rice intensification (SRI) cropping system can be implemented by partner farmers. partner farmer groups become independent in carrying out their farming activities using assembled technology that has been practiced

Chemical composition of Artemesia herba-alba essential oil and its larvicidal and pupicidal effects against Culex pipiens (Diptera; Culicidae)

Artemisia herba-alba Asso (A. herba-alba) (Asteraceae) is widely used in herbal medicine as it is a mine of natural molecules such as davanone, which is an interesting product on the international market. The present research proposes a method for controlling the late larval (L4) and pupal stages of Culex pipiens based on the essential oil of A. herba-alba. The aerial part of this plant was extracted by hydrodistillation and then analyzed by gas chromatography coupled with mass spectrometry (CPG/SM) to determine its chemical composition. Three concentrations (1, 5 and 10 μl/ml) were prepared and directly tested on larvae (L4) and pupae under laboratory conditions to measure LC50, LC90, LT50, and LT90 values. The yield obtained in this study was 1.5%. Further, the analysis showed that the oil of A. herba-alba is a davanone chemotype that consists mainly of davanone (48.8%). The efficiency of this essential oil for toxicological parameters (LC50 and LC90) were 3.278 μl/ml and 7.573 μl/ml for larvae, and 1.213 μl/ml and 2.288 μl/ml for pupae. This study indicates that the essential oil of A. herba-alba has toxic properties for Cx. pipiens larvae and pupae. These results are encouraging and open up exciting and promising horizons for its application in the production of bioinsecticides.

Conidia Production of Beauveria Bassiana in Solid Substrate Fermentation Using a Biphasic System

Beauveria bassiana is an important entomopathogenic fungus that is widely used as a bioinsecticide around the world. Conidia production is a critical step in the production of high-quality bioinsecticide. This study investigated three liquid culture mediums and five combinations of solid substrates to enhance conidia production by B. bassiana. The fungus was isolated from infected insects in the cocoa plantation of PT. Perkebunan Nusantara XII in Kediri, East Java, Indonesia. The three culture mediums were malt extract broth (MB), potato dextrose broth (PDB), and yeast and malt extract broth (YMB). Five combinations of solid substrate were used: 100% rice, 100% maize, 75%:25% rice:maize, 50%:50% rice:maize, and 25%:75% rice:maize. The biphasic system was used in this study, in which the fungus was first grown under submerged conditions and then was allowed to conidiate in solid-state conditions. The data showed that PDB was the optimum culture medium to produce blastophore and beauvericin, the active compound that acts as a mycoinsecticide. In the selection test, 100% rice was the optimum solid substrate to produce high amounts of conidia, and the consistency and production tests yielded the same results, with conidia counts of 1.93x109, 1.78x109, and 2.08x109, respectively. In a rice storability test, B. bassiana conidia numbers remained stable for up to 105 days of storage at room temperature.
 Keywords: Beauveria bassiana, culture medium, solid-substrate, conidia, biphasic system

Bacillus thuringiensis strain QBT220 pBtoxis plasmid structural instability enhances δ-endotoxins synthesis and bioinsecticidal activity

Bacillus thuringiensis subsp. israelensis (Bti) spherical parasporal crystal contains several insecticidal proteins used as environmentally safe alternative to toxic chemical pesticides. The exploration of a Bti strain isolated from Qatar QBT220 genes encoding the δ-endotoxins responsible of the insecticidal activities revealed the alteration of a 14-kb DNA region including the δ-endotoxins cry10A and cyt1C genes of pBtoxis plasmid. The presence of all the insecticidal genes except cry10A and cyt1C was explained by a structural instability of the plasmid pBtoxis. However, when compared with the Bti reference strains H14 and QBT217 that carry all δ-endotoxins coding genes, it was found that QBT220, has a significantly higher insecticidal activity against the dipteran insect Aedes aegypti larvae despite of the plasmid pBtoxis structural instability due to the alteration of cry10A and cyt1C genes. In addition, QBT220 showed the highest δ-endotoxin synthesis per spore, compared with that of the wildtype strains. These findings confirm that the altered genes cry10A and cyt1C are not mandatory for Bti insecticidal activities and on the other hand show a possible inhibitory effect played by the 2 proteins Cry10A and Cyt1C on the insecticidal activities of the other insecticidal proteins. In addition, the QBT220 increased δ-endotoxins synthesis per cell, makes this strain a good candidate for possible applications in the industrial production of bioinsecticides.

Isolation and Use of Bacillus thuringiensis for the Production of Bio-Insecticide in Control of Mosquito Larvae

Bacillus thuringiensis is a widely studied bacterium and it is known for its use in pest management. It is selectively active on pests and less likely to cause resistance; hence it is considered a suitable replacement to chemical pesticides. The study assessed the potential of Bacillus thuringiensis in controlling mosquito larvae. Bacillus thuringiensis isolates selected were tested against secondary stage larvae of mosquito. Thirty-six larvae (6 each) were transferred into each test tubes (7 x 9) cm with 30ml sterile distilled water. The stock suspension of cultures of Bacillus thuringiensis in broth was diluted to 107, 106, 10 5, 104, 103 and 102 in sterile water, following the McFarland standard method for microbial load count. The test tubes were kept at room temperature, larval mortality was observed over time within 24hrs. The results showed that all mosquito larvae died at the 107 and 106 dilutions but at dilutions 105, 104 and 103 though affecting mosquito larvae, it was highly dependent on time because there was a decrease in concentration. The study showed that B. thuringiensis is safe for use in aquatic environments, including drinking-water reservoirs, for the control of mosquito, black fly and nuisance insect larvae. The products should contain the ICPs and be free from other microorganisms and biologically active metabolites.

Development of a cost-effective medium for Photorhabdus temperata bioinsecticide production from wastewater and exploration of performance kinetic

This study investigates the optimization of the culture conditions for enhancing Photorhabdus temperata biopesticide production using wastewater (WS4) as a raw material. Box-Behnken design (BBD) was used to evaluate the effects of carbon to nitrogen ratio (C/N), sodium chloride concentration and inoculum size on P. temperata biomass production and insecticidal activity. For an enhanced biopesticide production, the optimum operating conditions were as follows: inoculum size = 4%; C/N ratio = 12.5 and [NaCl] = 4 g/L for two responses. 1.95 and 2.75 fold improvements in oral toxicity and biomass production were respectively obtained in the cost-effective medium developed in this study (WS4 I) using the three variables at their optimal values. Under the optimized conditions, WS4 I-grown cells exhibited higher membrane integrity according to flow cytometry analysis since dead cells presented only 9.2% compared to 29.2% in WS4. From batch fermentations carried out in WS4 I and WS4, P. temperata kinetic parameters in terms of biomass production and substrate consumption rates were modeled. The obtained results showed that the maximum specific growth rate in WS4 I was of 0.43 h−1 while that obtained in WS4 was of 0.14 h−1. In addition, the efficiency of P. temperata to metabolize organic carbon was enhanced by optimizing the culture conditions. It reached 72.66% instead of 46.18% in the control fermentation after 10 h of incubation. Under the optimized conditions, P. temperata cells showed the highest specific consumption rate resulting in a toxin synthesis improvement.

Acidic pretreatment as a chemical approach for enhanced Photorhabdus temperata bioinsecticide production from industrial wastewater

The chemical treatment of the wastewater used for the bioinsecticide production by the bacterium Photorhabdus temperata was investigated in this study. An improvement of the volatile suspended solids (VSS) solubilization along with an increase in protein, carbohydrate, reducing sugar and nitrogen concentrations were demonstrated after alkali and thermo-alkali hydrolysis. In contrast, the application of acidic and thermo-acidic pretreatments reduced the organic matter hydrolysis. Compared to untreated wastewater, the chemical oxygen demand (COD) solubilization and the heavy metal concentration, except manganese, were enhanced in all the chemically pretreated wastewaters. Although its low contribution in the solubilization of the wastewater organic matter, the acidic-pretreated wastewater showed the highest performance in supporting P. temperata biopesticide production. Indeed, using the acidic-pretreated wastewater as a fermentation medium decreased the lag phase, enhanced the growth of the strain K122 to reach a final biomass production of 20 × 108 cells/mL, increased culturable cell count to 262 × 106 cells/mL and improved oral toxicity against Ephestia kuehniella larvae by 68.4%. Among chemical pretreatments performed, the acidic hydrolysis was demonstrated to be the unique promising one for P. temperata bioinsecticide production due to its ability to reduce aromatic compounds as shown by Gas Chromatography-Mass Spectrometry (GC-MS) analysis.

Bacillus thuringiensis strains isolated from Qatari soil, synthesizing δ-endotoxins highly active against the disease vector insect Aedes aegypti Bora Bora

Bacillus thuringiensis (Bt) is a Gram-positive soil bacterium that has been recognized as an effective bioinsecticide active against plant, animal and human pathogenic and disease vector insects. During its sporulation phase, Bt produces crystals consisting of δ-endotoxins, which upon ingestion kill specifically insect larvae. Bt subsp. israelensis (Bti) is very active against dipteran insects. Bti based bioinsecticides are considered as a sustainable solution to control the Dipteran insects responsible of plant, animal and human diseases. In this study, Bti strains isolated from Qatar soil were analyzed for their insecticidal activities against the dipteran insect Aedes aegypti Bora Bora (Culicidae, Diptera) and for their δ-endotoxins yields per cell. Among the local Bti strains, four exceptional strains producing spherical crystals, were found to be more insecticidal than the reference strain Bti H14. When tested for their δ-endotoxin yield, the Bti QBT217 strain, producing typical spherical crystals and having the best insecticidal activity, was recognized as the best candidate strain for potential bioinsecticide production and biological control of dipteran insects, particularly the disease vector insect A. aegypti.

The Production of Bioinsecticide Based From Pong-Pong Fruit Seed Extract by Ultrasonic Waved Extraction Using NADES Solvent

The Production of Bioinsecticide Based From Pong-Pong Fruit Seed Extract by Ultrasonic Waved Extraction Using NADES Solvent

Towards a new biological control approach for Photorhabdus temperata bioinsecticide production through the bioconversion of Tunisian industrial wastewater

A novel bioconversion approach of Tunisian wastewater to low-cost Photorhabdus temperata bioinsecticide is presented in this study. Our results showed that when cultured on the food industry wastewater (WS4), P. temperata cells exhibited oral toxicity of about 42%, which is the same as those cultured in complex medium (CM), used as control. Moreover, variants small colony polymorphism (Vsm) of the strain K122 was completely avoided after a prolonged incubation. However, viable but non-culturable (VBNC) state was enhanced with the maximum colony-forming units (CFU) count of 9 × 106 cells/mL obtained after 48 h of incubation in the WS4. According to flow cytometry analysis, almost 100% of P. temperata cells were viable until 48 h of incubation. The appearance of propidium iodide (PI) positively stained cells was observed after a prolonged incubation with a maximum of 17% of damaged cells in WS1. In order to follow the progress of P. temperata fermentation process carried out in industrial wastewater, we established for the first time, the mathematical relationship between total cell counts, CFU counts and oral toxicity of P. temperata strain K122. Indeed, irrespective of the medium used, the relationship between CFU count and total cell count followed a power law. Additionally, when plotting CFU count, or total cell count against toxicity, a semi-log linear relationship was obtained. Our results proved the efficiency of this bioconversion approach to produce bioinsecticide based on the entomopathogenic bacterium P. temperata, with practical benefits in terms of cost production and wastewater management.

Effectiveness of Beauveria bassiana Vuill. isolate on various culture media and its pathogenicity against Tribolium castaneum

One of the selected microbes that has been widely used for production of bioinsecticides is B. bassiana. The fungus is very effective in suppressing larvae of pest insects Lepidoptera and Coleoptera. However, data on the potential of B. bassiana as bioinsecticides against Coleoptera are not widely available in Indonesia. This study aims to determine the effectiveness of B. bassiana against warehouse pests from Coleoptera Tribolium castaneum. Culture of B. bassiana on Potato Dextrose Agar media was propagated on corn media, corn liquid media, PDA media, and rice media. Tribolium castaneum test insects are derived from the remains of rice mills and then kept until imago lay eggs and have a uniform age. Then B. Bassiana was applied to Tribolium castaneum. Observations were made every 24 hours on the mortality of Tribolium castaneum. The results showed that rice media is the best media used for propagation of Beauveria bassiana when viewed from spore density where the average spore density is 12.53 X 106. The fungus isolates that were propagated using liquid corn media, rice media, and corn media were the best isolates that could be used to kill T. castaneum pest in warehouse.

Characteristics of Bacillus thuringiensis isolates indigenous soil of South Sumatra (Indonesia) and their pathogenicity against oil palm pests Oryctes rhinoceros (Coleoptera: Scarabaeidae)

Abstract. Pujiastuti Y, Arsi, Sandi S. 2020. Characteristics of Bacillus thuringiensis isolates indigenous soil of South Sumatra (Indonesia) and their pathogenicity against oil palm pests Oryctes rhinoceros (Coleoptera: Scarabaeidae). Biodiversitas 21: 1287-1294. Bacillus thuringiensis is a gram-positive, entomopathogenic bacterium that could be isolated from soil and be used to control various plant pests. Oryctes rhinoceros is an important pest in oil palm. Application of B. thuringiensis-based bioinsecticides is an alternative in controlling these pests. The purposes of this study were to isolate and identify B. thuringiensis bacteria from the soil of South Sumatra, production of B. thuringiensis-based bioinsecticides and to test their toxicity to O. rhinoceros larvae. The study was conducted in several cities/districts in the province of South Sumatra. Soil samples were taken from various habitats and B. thuringiensis isolates were grown on NGKG agar media. Among 76 soil samples (6 districts and 2 cities) B. thuringiensis colonies were obtained leading to 24 isolates of B. thuringiensis. Toxicity screening tests for armyworm Spodoptera litura were 55.79% (53 isolates) and their mortality to 25.26% O. rhinoceros larvae (24 isolates). From these isolates whose effectively killed O. rhinoceros larvae, 10 isolates were taken and propagated with Nutrient Broth (NB) and biourine enriched with 5% molasses. Number of spores produced was counted during 24, 48 and 72 hours. Furthermore, a bioassay test was carried out on O. rhinoceros larvae for 7 days. Isolate of KJ3P1 caused the highest mortality of O. rhinoceros larvae after 7 days of observation. SDS Page resulted in KJ3P1 and KJ3R5 isolates showing several bands whose content of various types of protein molecular weight. Isolation of B. thuringiensis in South Sumatra produced 2 isolates potentially to be active ingredients in production of bioinsecticides which were effective in killing O. rhinoceros larvae.

Optimization of bio-insecticide production by Tunisian Bacillus thuringiensis israelensis and its application in the field

Bacillus thuringiensis israelensis biological larvicides represent a good approach to support vector borne diseases control programs targeting earlier aquatic larval stages to limit their adult mosquito spreading.BUPM98, a Tunisian B. thuringiensis israelensis strain, is highly active against Aedes aegypti larvae. For the production of BUPM98 bioinsecticide, an agricultural by-product medium consisting of gruel and soybean meal has been optimized. Optimization of the medium composition was performed with statistical methodology based on experimental designs. Accordingly, the optimum toxin production was 2375.36 ± 48.52 mg/L at the following conditions: K2HPO4: 60 mM, pH: 7.6 and soybean meal: 32 g/L, which led to an improvement of 30% compared to the starting conditions. The impact of water quality on the effectiveness of BUPM98 delta-endotoxins against Culex pipiens larvae was investigated. Indeed, under field conditions, the direct application of BUPM98 fermentation product in stagnant water pools reduced the immature density of C. pipiens larvae by more than 95% after 48 h of treatment. Prolonged residual action was observed until 14 days. In conclusion, BUPM98, a local strain of B. thuringiensis israelensis, is very promising bioinsecticide for mosquito population management in Tunisia.

Genome sequence analysis of a novel Bacillus thuringiensis strain BLB406 active against Aedes aegypti larvae, a novel potential bioinsecticide

BLB406 is a novel isolate of Bacillus thuringiensis with a larvicidal activity against Aedes aegypti larvae. It displays original plasmidic and crystal protein patterns.The present work reported molecular and bioinformatic analyses for the genome sequence of BLB406 using MiSeq Illumina next-generation sequencing technology. The reads were assembled by Velvet tool. Using RAST program and PGAAP, the genome of BLB406 strain was shown to contain 6297 genes corresponding to 5924 protein coding sequences. The BLB406 genome investigation with BtToxin_scanner program shows that this strain has an original and different combination of toxins compared to the published ones: five cry genes (cry11, cry22, cry2, cry60, cry64) and two distinct vegetative insecticidal vip4 genes. This combination provides a potential larvicidal and anti-cancer activities to BLB406. It might be a potential solution to some problems such as the narrow insecticidal spectra and insect resistance. The whole BLB406 genome information provides a valuable background for future in silico analyses as well as biotechnological applications in order to increase the production of commercial bioinsecticide based on BLB406 B. thuringiensis strain.

The Effect of NADES Ratio Solvent and Sonication Time in Extraction of Papaya Sap for Bio-insecticide Production

Along with the increasing quantity of agriculture in Indonesia, should be followed by improving the quality of agriculture. Generally one of the indicators or parameters commonly used to indicate the quality of agriculture is the number of pests that damage plants in certain areas. The control of disturbing organisms on the plant can be minimized by the bio-insecticide production of Carica papaya which is tapped and extracted with ultrasonic-assisted using a mixed NADES solvent from Cholin chloride (ChCl) and Oxalic acid (Ox) with varying sonification time for 10, 20, 30 minutes and solvent ratio of 1: 2 and 2: 1. Bio-insecticide was produced by conducting qualitative enzyme activity of with lowry method and quantitatively using UV / Vis spectrophotometer aid with 750 nm wavelength and efficacy test. The best extraction result was obtained at NADES solvent with 1: 2 ratio with extraction time for 30 min with extraction time for 30 minutes yielding protein concentration equal to 7,23 ppm and tyrosine concentration equal to 7,9 ppm. Bio-insecticide samples effectively kill grayak caterpillars with 100% mortality within 5-7 days.

Production of Plutella xylostella bioinsecticide for brassicaceae plant based on bromelain enzyme extracted from pineapple peel and isothiocyanate extracted from broccoli stem and radish peel

Brassicaceae plant is a commodity of edible plant that is very important for Indonesia, with cabbage worm (Plutella xylostella) as its main pest. Pest control with biopesticide is needed to preserve environment and increase crops production. Several biological substance is toxic to cabbage worm, including bromelain enzyme and isothiocyanates. Extraction of bromelain is experimented by varying mixing duration (15, 30, 45, 60 mins) and solvent type (distilled water and phosphate buffer). Extraction of isothiocyanate is experimented by varying the solvent ratio (1:1, 1:2, 1:3), using dichloromethane as solvent. For the broccoli stem sample, phosphate buffer is added to maintain pH. Enzyme activity test is used with product tyrosin concentration as standard. Isothiocyanate is determined with GC-MS analysis. The most effective extraction method for bromelain is with distilled water as solvent with 15 min mixing time. The most effective extraction method for isothiocyanate is with 1:1 solvent ratio at 37°C temperature without adding phosphate buffer. Efficacy of each sample to cabbage worm is also experimented by feeding it broccoli leaf spread with sample and observing the mortality rate. Bromelain and isothiocyanate proved to be toxic to cabbage worm and can be used as alternative bioinsecticide with the highest mortality of cabbage worm is by isothiocyanate from radish peel, reaching 100%.