Production Of Biopesticides Research Articles

Development of point-of-need colourimetric, isothermal diagnostic assays for specific detection of Bacillus subtilis using shikimate dehydrogenase gene.

The largest obstacle in the promotion of biopesticides is the existence of counterfeit products available in the market. Identification and quantification of antagonistic organisms in biopesticide products are the key to the reduction of spurious microbial pesticides. In this study, we have developed a simple, sensitive, isothermal-based colourimetric assay for specific detection of Bacillus subtilis from the biopesticide formulations and soil samples. A region specific to B. subtilis which codes for shikimate dehydrogenase was identified through in silico analysis. We employed conventional PCR, loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and qPCR for specific detection of B. subtilis in soil samples and biopesticide formulations. Specificity tests showed that the PCR primers amplified an amplicon of 521bp in four strains of B. subtilis only, and no amplification was found in negative control samples. Similarly, the LAMP assay showed sky blue colour in all four strains of B. subtilis and violet colour in negative control samples. Whereas in the RPA assay, upon the addition of SYBR Green dye, a bright green colour was seen in B. subtilis strains, while a brick-red colour was observed in negative control samples by visualizing under a UV transilluminator. The qPCR assay showed specific amplifications with a Ct value of 12 for B. subtilis strains and no amplification in negative control samples. In the sensitivity test, PCR could amplify DNA of B. subtilis up to 500pg/µL. DNA concentration as low as 10pg/µL was enough to show the colour change in the LAMP as well as the RPA assays, whereas the qPCR assay showed sensitivity till 100pg/µL. All four diagnostic assays developed in the study have been validated in soil samples and B. subtilis-based biopesticides. Compared to conventional PCR, the qPCR assay has the advantage of quantification and visualizing the result in real-time, whereas LAMP and RPA assays have the benefits of being colourimetric and less time-consuming. The other advantages are that the results can be visualized with the naked eye, and these assays do not require a costly thermal cycler and gel documentation system. Hence, LAMP and RPA assays are highly suitable for developing point-of-need diagnostic kits and, in turn, help regulators assess the quality of biopesticides in the market.

Microbial Production of Biopesticides for Sustainable Agriculture

Food security is threatened by biotic stress on crops, e.g., from invasive pests, in the context of climate change. Pest control has traditionally been achieved using synthetic pesticides, yet pollution and the persistence and toxicity of some pesticides are inducing food contamination and, in turn, generating public concern. As a consequence, biopesticides are increasingly used, notably for organic crops. For instance, some microorganisms produce biopesticidal compounds such as secondary metabolites or growth-inhibitory enzymes. Most reviews on this topic mainly focus on describing microbial species and their active compounds. However, there are very few studies and reviews describing various process parameters. Here, we review both microbial biopesticides and factors controlling physicochemical conditions for the scaling up of biopesticide production. We present biopesticides from bacteria (e.g., Bacillus thuringiensis), algae, fungi, and viruses and review the biocontrol mechanisms and applications of commercial biopesticides. The benefits of genetic engineering for enhancing activity and drawbacks such as commercialization are also discussed.

An economic analysis of pattern of investment for biopesticides production unit establishment in Chhattisgarh

An economic analysis of pattern of investment for biopesticides production unit establishment in Chhattisgarh

Microbial pesticides – challenges and future perspectives for testing and safety assessment with respect to human health

Plant protection measures are necessary to prevent pests and diseases from attacking and destroying crop plants and to meet consumer demands for agricultural produce. In the last decades the use of chemical pesticides has largely increased. Farmers are looking for alternatives. Biopesticides should be considered a sustainable solution. They may be less toxic than chemical pesticides, be very specific to the target pest, decompose quickly, and be less likely to cause resistance. On the other hand, lower efficacy and higher costs are two disadvantages of many biopesticides. Biopesticides include macroorganisms, natural compounds and microorganisms. Microbial pesticides are the most widely used and studied class of biopesticides. The greatest difference between microbial and chemical pesticides is the ability of the former to potentially multiply in the environment and on the crop plant after application. The data requirements for the European Union and the United States Environmental Protection Agency are highlighted, as these regulatory processes are the most followed in regions where local regulations for biopesticide products are not available or vague. New Approach Methods already proposed or harmonized for chemical pesticides are presented and discussed with respect to their use in evaluating microbial pesticide formulations. Evaluating the microbials themselves is not as simple as using the same validated New Approach Methods as for synthetic pesticides. Therefore, the authors suggest considering New Approach Method strategies specifically for microbials and global harmonization with acceptability with the advancements of such approaches. Further discussion is needed and greatly appreciated by the experts.

Comparison of the performance of multiple whole-genome sequence-based tools for the identification of Bacillus cereus sensu stricto biovar Thuringiensis.

The Bacillus cereus sensu stricto (s.s.) species comprises strains of biovar Thuringiensis (Bt) known for their bioinsecticidal activity, as well as strains with foodborne pathogenic potential. Bt strains are identified (i) based on the production of insecticidal crystal proteins, also known as Bt toxins, or (ii) based on the presence of cry, cyt, and vip genes, which encode Bt toxins. Multiple bioinformatics tools have been developed for the detection of crystal protein-encoding genes based on whole-genome sequencing (WGS) data. However, the performance of these tools is yet to be evaluated using phenotypic data. Thus, the goal of this study was to assess the performance of four bioinformatics tools for the detection of crystal protein-encoding genes. The accuracy of sequence-based identification of Bt was determined in reference to phenotypic microscope-based screening for the production of crystal proteins. A total of 58 diverse B. cereus sensu lato strains isolated from clinical, food, environmental, and commercial biopesticide products underwent WGS. Isolates were examined for crystal protein production using phase contrast microscopy. Crystal protein-encoding genes were detected using BtToxin_Digger, BTyper3, IDOPS (identification of pesticidal sequences), and Cry_processor. Out of 58 isolates, the phenotypic production of crystal proteins was confirmed for 18 isolates. Specificity and sensitivity of Bt identification based on sequences were 0.85 and 0.94 for BtToxin_Digger, 0.97 and 0.89 for BTyper3, 0.95 and 0.94 for IDOPS, and 0.88 and 1.00 for Cry_processor, respectively. Cry_processor predicted crystal protein production with the highest specificity, and BtToxin_Digger and IDOPS predicted crystal protein production with the highest sensitivity. Three out of four tested bioinformatics tools performed well overall, with IDOPS achieving high sensitivity and specificity (>0.90).IMPORTANCEStrains of Bacillus cereus sensu stricto (s.s.) biovar Thuringiensis (Bt) are used as organic biopesticides. Bt is differentiated from the foodborne pathogen Bacillus cereus s.s. by the production of insecticidal crystal proteins. Thus, reliable genomic identification of biovar Thuringiensis is necessary to ensure food safety and facilitate risk assessment. This study assessed the accuracy of whole-genome sequencing (WGS)-based identification of Bt compared to phenotypic microscopy-based screening for crystal protein production. Multiple bioinformatics tools were compared to assess their performance in predicting crystal protein production. Among them, identification of pesticidal sequences performed best overall at WGS-based Bt identification.

Bioenergy and Biopesticides Production in Serbia—Could Invasive Alien Species Contribute to Sustainability?

The critical role of energy in contemporary life and the environmental challenges associated with its production imply the need for research and exploration of its novel resources. The present review paper emphasizes the continuous exploitation of non-renewable energy sources, suggesting the transition toward renewable energy sources, termed ‘green energy’, as a crucial step for sustainable development. The research methodology involves a comprehensive review of articles, statistical data analysis, and examination of databases. The main focus is biomass, a valuable resource for bioenergy and biopesticide production, highlighting not only its traditional diverse sources, such as agricultural waste and industrial residues, but also non-edible invasive alien plant species. This study explores the utilization of invasive alien species in circular economy practices, considering their role in bioenergy and biopesticide production. The potential conflict between bioproduct acquisition and food sector competition is discussed, along with the need for a shift in approaching non-edible biomass sources. The paper emphasizes the untapped potential of under-explored biomass resources and the necessity for policy alignment and public awareness. Species with a significant potential for these sustainable strategies include Acer negundo L., Ailanthus altisima (Mill.) Swingle., Amorpha fruticosa L., Elaengus angustifolia L., Falopia japonica (Houtt.) Ronse Decr., Hibiscus syriacus L., Koelreuteria paniculata Laxm., Paulownia tomentosa Siebold and Zucc., Partenocissus quenquefolia (L.) Planch., Rhus typhina L., Robinia pseudoacacia L. and Thuja orientalis L. In conclusion, the paper highlights the intertwined relationship between energy, environmental sustainability, and circular economy principles, providing insights into Serbia’s efforts and potential in adopting nature-based solutions for bioenergy and biopesticides acquisition.

Phytochemical characterisation and toxicity effect of Tithonia diversifolia (Hemls.) A. Gray leaf extract on fall army worm Spodoptera frugiperda (JE Smith) larvae.

To mitigate the losses due to fall armyworm (FAW) infestation in maize, chemical pesticides had been the first choice and widely used as an emergency response. However, it comes with attendant health effect. This necessitates the development of plant based effective and safer pesticides. This research investigate response of fall armyworm larvae when they are exposed to crude and partially purified Tithonia diversifolia leaf extract. Chemical constituent of the extract was identified using NIST08.LIB library spectra provided by the software on a GC-MS system and FTIR analysis was done using KBr pellet technique with a resolution and scanning speed of 4 cm-1 and 2 mm/s. Dose dependent toxicity assay of T.diversifolia extracts on FAW at different growth stages under controlled environment in laboratory, followed by its effect under phytotron were examined against control and azadirachtin from neem. The GC-MS of the butanol eluent revealed 20 compounds out of which the major ones being beta-d-glucopyranoside, methyl (15.225 %) palmitic acid, TMS derivative (10.98 %) and hexadecanoic acid, 2-[(trimethylsily)oxy]-, methyl ester (8.75 %). The FT-IR spectroscopic analysis of the butanol eluent of T.diversifolia leaf extract revealed the presence of alcohols, phenols, aldehydes, ketones, alkanes and primary amines. The butanol eluent and crude extract caused 96 % mortality at neonate and first instar FAW larvae. The toxic and repellant effects revealed by diet bioassay and phytotron experiment respectively suggest that butanol eluent of T.diversifolia leaf extract could be a good and effective target for biopesticide production against FAW.

Exploring biorefinery alternatives for biowaste valorization: a techno-economic assessment of enzymatic hydrolysis coupled with anaerobic digestion or solid-state fermentation for high-value bioproducts

ABSTRACT Enzymatic hydrolysis of organic waste is gaining relevance as a complementary technology to conventional biological treatments. Moreover, biorefineries are emerging as a sustainable scenario to integrate waste valorization and high-value bioproducts production. However, their application on municipal solid waste is still limited. This study systematically evaluates the techno-economic feasibility of the conversion of the organic fraction of municipal solid waste (OFMSW) into high-value bioproducts through enzymatic hydrolysis. Two key variables are examined: (a) the source of the enzymes: commercial or on-site produced using OFMSW, and (b) the treatment of the solid hydrolyzate fraction: solid-state fermentation (SSF) for the production of biopesticides or anaerobic digestion for the production of energy. As a result, four different biorefinery scenarios are generated and compared in terms of profitability. Results showed that the most profitable scenario was to produce enzymes on-site and valorize the solid fraction via SSF, with an internal rate of return of 13%. This scenario led to higher profit margins (74%) and a reduced payback time (6 years), in contrast with commercial enzymes that led to an unprofitable biorefinery. Also, the simultaneous production of higher-value bioproducts and energy reduced the economic dependence of OFMSW treatment on policy instruments while remaining energetically self-sufficient. The profitability of the biorefinery scenarios evaluated was heavily dependent on the enzyme price and the efficiency of the anaerobic digestion process, highlighting the importance of cost-efficient enzyme production alternatives and high-quality OFMSW. This paper contributes to understanding the potential role of enzymes in future OFMSW biorefineries and offers economical insights on different configurations.

Production of biopesticide from Melia azedarach Linn extract obtained by supercritical fluid extraction for the control of Tetranychus urticae

Compounds derived from plant extracts, such as those from the fruits and leaves of Melia azedarach Linn (chinaberry tree), which contain azadirachtin, have been utilized for crop pest control. However, these natural compounds are susceptible to degradation during extraction and in field conditions. To address this, extraction processes involving supercritical fluid and microencapsulation via spray drying may be employed to obtain and preserve biopesticidal molecules. This study focuses on producing a biopesticide from M. azedarach L. fruit extract using supercritical CO2, and microencapsulation via spray drying with a blend of cheese whey and gum Arabic. The extract was obtained from chinaberry fruits rich in azadirachtin (736.92 μg/L) using supercritical CO2 at 250 bar and 50 °C for 120 min, with a yield of 3.44%. The extract was then encapsulated using cheese whey and gum Arabic (70:30) as encapsulating agents at extract:encapsulating agents ratios of 1:10 and 1:15. Mortality rates of Tetranychus urticae Koch adult females and their eggs, in contact and residual assays with both non-encapsulated and encapsulated extract, did not significantly differ (p > 0.05) from the commercial acaricide abamectin. This study introduces a potential new biopesticide that utilizes chinaberry extract obtained through supercritical CO2 extraction and encapsulation for industrial scale-up studies, and this approach holds promise for the production of an effective biopesticide for crops affected by two-spotted spider mites.

Diverse Approaches to Insect Control: Utilizing Brassica carinata (A.) Braun and Camelina sativa (L.) Crantz Oil as Modern Bioinsecticides

The forest environment is exposed to a number of harmful factors that significantly reduce the resistance of forest stands, often leading to their extinction. In addition to abiotic and anthropogenic factors, biotic factors pose a significant threat to forests, among which insect pests are at the top of the list. Until now, the use of chemical insecticides has been considered the most effective method of pest control, resulting in pesticide residue in the environment. In an effort to minimize the harmful effects of insecticides, the European Union (EU), through EU Commission Implementing Regulations 2022/94, 2021/2081, 2021/795, and 2020/1643, has decided to withdraw from use a number of preparations containing compounds such as phosmet, indoxacarb, alpha-cypermethrin, and imidacloprid, among others. Botanical insecticides appear to be a promising alternative. Among them, plant oils and essential oils have become an innovative solution for controlling pests not only of forests but also of agricultural crops. The purpose of this literature review was to select oilseed plants with great biological potential. The rich chemical compositions of the seeds of Brassica carinata (A.) Braun and Camelina sativa (L.) Cranz predispose them to use as raw materials for the production of biopesticides with broad mechanisms of action. On the one hand, the oil will provide a physical action of covering pests feeding on a plant with a thin film, which will consequently lead to a reduction in gas exchange processes. On the other hand, the bioactive compounds in it or extracts of fat-insoluble compounds suspended in it and derived from the pomace fraction may have deterrent or lethal effects. This paper presents evidence of the potential action of these raw materials. Preparations based on these oils will not pose a threat to living beings and will not negatively affect the environment, thus allowing them to gain social acceptance.

The role of agro-input dealer certification in promoting sustainable pest control: insights from Uganda

ABSTRACT Pesticides are crucial for increasing agricultural productivity, but they have also been linked to a range of health and environmental risks. In this paper, we used nationally representative data from 557 agro-dealers in Uganda to assess the role of agro-dealer certification in improving knowledge and practices related to environmentally-friendly pest control. We found that almost half of the sampled agro-dealers were not certified or accredited by regulatory bodies, even though this is a prerequisite for selling pesticides in the country. Results further showed that only 16% of the agro-input shops were selling biopesticide products, largely due to a lack of awareness, access and demand from farmers. Regression results showed that certified agro-dealers were 9–12 percentage points more likely to know about biopesticides and integrated pest management, and 8–10 percentage points more likely to sell biopesticide products, compared to their non-certified counterparts. Our findings imply that agro-dealer certification courses can play an important role in raising knowledge and stimulating the supply of environmentally-benign pest control products. We identified regulatory enforcement, a decentralized certification system and agro-dealer associations as some of the potential pathways for incentivising compliance with certification requirements, thereby promoting lower-risk pest control products and strategies.

A Comprehensive Study of MicroRNA in Baculoviruses.

Baculoviruses are viral pathogens that infect different species of Lepidoptera, Diptera, and Hymenoptera, with a global distribution. Due to their biological characteristics and the biotechnological applications derived from these entities, the Baculoviridae family is an important subject of study and manipulation in the natural sciences. With the advent of RNA interference mechanisms, the presence of baculoviral genes that do not code for proteins but instead generate transcripts similar to microRNAs (miRNAs) has been described. These miRNAs are functionally associated with the regulation of gene expression, both in viral and host sequences. This article provides a comprehensive review of miRNA biogenesis, function, and characterization in general, with a specific focus on those identified in baculoviruses. Furthermore, it delves into the specific roles of baculoviral miRNAs in regulating viral and host genes and presents structural and thermodynamic stability studies that are useful for detecting shared characteristics with predictive utility. This review aims to expand our understanding of the baculoviral miRNAome, contributing to improvements in the production of baculovirus-based biopesticides, management of resistance phenomena in pests, enhancement of recombinant protein production systems, and development of diverse and improved BacMam vectors to meet biomedical demands.

Sustainable Production and Application of Bt Biopesticides

Sustainable Production and Application of Bt Biopesticides

Economic analysis of major constraints of production and marketing of biopesticides in Chhattisgarh

Economic analysis of major constraints of production and marketing of biopesticides in Chhattisgarh

Policy options for promoting wider use of biopesticides in Thai agriculture

Biopesticides are rapidly growing in importance for crop protection globally, but nearly all growth is happening in high income countries. No previous work systematically analyzed how lower-income countries can better benefit from the increased availability of biopesticides, which is important because these countries are particularly affected by the adverse effects of chemical pesticide use. Here we review the legislation of biopesticides in Thailand and combine this with stakeholder interviews and interview data from 300 smallholder farmers producing rice, fruit, vegetables, and flowers. We find that Thailand has adopted a biopesticide registration system that facilitates a fast-track registration, but it is still relatively costly considering the small market size. While 65 % of the sampled farmers used biopesticides, most farmers still heavily relied on conventional pesticides as their main method to control pests. Education, farming experience, positive attitudes of biopesticides, adoption of other integrated pest management methods and contacts with government extension agents were positively associated with biopesticide use. Coordinated action is needed to stimulate the supply of a wider range of biopesticide products while promoting adoption among farmers.

Evaluation of the Essential Oils Used in the Production of Biopesticides: Assessing Their Toxicity toward Both Arthropod Target Species and Beneficial Pollinators

Biopesticides, alongside the expansive growth of conventional agriculture, emerged as a hopeful avenue for mitigating the environmental impact of synthetic chemicals. Essential oils derived from various plant species are regarded as environmentally friendly and have been suggested by numerous researchers for combating the pest species. However, in addition to their detrimental effects on harmful species, these essential oils exert varying degrees of impact on non-target species with which they share ecological relationships. This review serves the dual purpose of updating data on the use of the essential oils as biopesticides, considering the substantial increase in research output in the recent years. Simultaneously, it aims to provide a focused synthesis on the essential oils currently approved for commercial use as biopesticides, and shedding light on their impact on beneficial pollinator species, which may inadvertently suffer from the application of substances deemed safe by most users.

Valorization of Chlorella thermophila biomass cultivated in dairy wastewater for biopesticide production against bacterial rice blight: a circular biorefinery approach

Biopesticides offer a sustainable and efficient alternative to synthetic pesticides, providing a safer and more eco-friendly solution to pest management. The present work proposes an innovative approach that integrates crop protection and wastewater treatment using thermophilic microalgal strain Chlorella thermophila (CT) cultivated in nutrient-rich dairy wastewater as a growth medium. The microalgae was cultivated mixotrophically and was able to reduce both organic carbon as well as nutrient load of the dairy wastewater efficiently. The integrated circular biorefinery approach combines biomass cultivation, extraction of biopesticide compounds, and conversion to biocrude. The antimicrobial activity of the biopesticidal extracts against Xanthomonas oryzae and Pantoea agglomerans, the causative agent of bacterial rice blight, is assessed through in vitro studies. The biomass extract obtained is able to inhibit the growth of both the above-mentioned plant pathogens successfully. Mass spectroscopy analysis indicates the presence of Neophytadiene that has previously been reported for the inhibition of several pathogenic bacteria and fungi. Several other value-added products such as linoleic acid and nervonic acids were also been detected in the microalgal biomass which have extremely high nutraceutical and medicinal values. Furthermore, the study investigates the potential for co-production of biocrude from the biorefinery process via hydrothermal liquefaction. Overall, the findings of this present work represent an innovative and sustainable approach that combines wastewater treatment and crop protection using microalgal biomass.

Training on making biopesticides as effort to strengthen organic agriculture by Taruna Tani Lestari

Pests are one of the problems in agriculture. The use of organic pesticides still needs to be developed in Gentungan Village. Training in making biopesticides is useful for controlling pests and diseases using environmentally friendly natural ingredients. In addition, to foster the ability of Taruna Tani Lestari in Gentungan Village to make biopesticides independently and disseminate them to local farmers. The implementation method involved presentations, questions and answers, practices, modules, and evaluation designs. The material presented included the meaning and benefits of biopesticides, the natural ingredients used, the correct dosage, and the stages of making biopesticides. After the training, the output obtained included members of Taruna Tani Lestari knowing the natural ingredients for making biopesticides, members of Taruna Tani Lestari understanding the benefits of using biopesticides, members of Taruna Tani Lestari being able to make biopesticides independently and educate local farmers and display biopesticide products.

Proposed Conceptual Framework for Studying Non-Users Rice Farmers' Intention to Use Commercial Biopesticides Products

Commercial biopesticide products have been developed for a long time in Indonesia, but until now the level of application is still relatively low. This paper aims to determine the factors that are expected to influence the intention of non-users rice farmers to use commercial biopesticide products. Furthermore, a conceptual framework of intention to use commercial biopesticide products is proposed. Based on the literature review of previous empirical studies in various contexts, this paper has explored the factors that influence the intention of using biopesticides. Unified Theory of Acceptance and Use of Technology 2 (UTAUT2) is employed as the main theoretical basis in this study which can be extended with relevant constructs to the research context (extended UTAUT2). The proposed conceptual framework recommends that non-users rice farmers' intention to use commercial biopesticide products is expected to be influenced by variables namely performance expectancy, effort expectancy, social influence, facilitating condition, price value, perceived need, information publicity, and product marketing. This integrative conceptual framework is expected to have a theoretical contribution to understand the factors of non-users rice farmers’ intention to use commercial biopesticide products. In addition, it is beneficial for policymakers and managers to formulate and implement strategies to increase rice farmers' intention to use commercial biopesticide products.

Trichoderma as a biological control agent: mechanisms of action, benefits for crops and development of formulations.

Currently, the food and economic losses generated by the attack of phytopathogens on the agricultural sector constitute a severe problem. Conventional crop protection techniques based on the application of synthetic pesticides to combat these undesirable microorganisms have also begun to represent an inconvenience since the excessive use of these substances is associated with contamination problems and severe damage to the health of farmers, consumers, and communities surrounding the fields, as well as the generation of resistance by the phytopathogens to be combated. Using biocontrol agents such as Trichoderma to mitigate the attack of phytopathogens represents an alternative to synthetic pesticides, safe for health and the environment. This work explains the mechanisms of action through which Trichoderma exerts biological control, some of the beneficial aspects that it confers to the development of crops through its symbiotic interaction with plants, and the bioremedial effects that it presents in fields contaminated by synthetic pesticides. Also, detail the production of spore-based biopesticides through fermentation processes and formulation development.