Bacillus thuringiensis isolated from soil in oil palm plantation with high toxicity against Oryctes rhinoceros larvae

Aims@#Oryctes rhinoceros beetle is one of the most damaging pests of oil palm and cause high oil palm mortality. The empty fruit bunch mulch and rotten old trunk of oil palm in the field provide the organic matter for the breeding sites and increases the number of O. rhinoceros larvae. Bacillus thuringiensis as bioinsecticide can synthesize crystal proteins toxic to the larvae. The present study was aimed to find effective B. thuringiensis isolates as biopesticide against O. rhinoceros larvae. @*Methodology and results@#Screening process was carried out through heating of soil sample suspension at 80 °C to eliminate the non-spore formers and plated onto T3 medium. Colony morphology was observed, followed by Gram and endospore staining. The crystal protein was observed by Coomassie Brilliant Blue (CBB) staining. Bioassay test was conducted by force-feed method followed by food contamination method. The results showed isolates SBB33 and SBB35 were able to infect and caused high mortality to the O. rhinoceros larvae. Isolates SBB33 and SBB35 showed the highest mortality against 1st instar larvae (94.44% and 75% respectively) and 3rd instar larvae (64.8% and 60% respectively) compared to control treatments. The 16S rRNA gene sequencing showed SBB33 has high similarity with B. thuringiensis strain 3S2-3, while SBB35 has high similarity with B. thuringiensis strain GCU_BTi10. Protein separation of the spore-crystal mixture by SDS-PAGE showed the prominence of 66 kDa protein band that was predicted to be Cry toxins which is specific to coleopterans insect.@*Conclusion, significance and impact of study@#Bacillus thuringiensis isolates SBB33 and SBB35 have high potential as biopesticides against O. rhinoceros larvae and could be used to control major pests in oil palm plantation.

Formulation of Insecticidal Nematode

Insecticidal nematodes or entomopathogenic nematodes (EPNs) in the genera Heterorhabditis and Steinernema are lethal obligate parasites of insect pests. These nematodes carry specific pathogenic bacteria which are released into the insect hemocoel after penetration of the insect host. The economic importance of entomopathogenic nematodes (EPNs) is increasing as the nematodes are amenable for mass production, formulation, handling and application on a large scale and is currently marketed worldwide for use. The objective of the review is to discuss the principles of formulation and quality control, latest development and future perspectives of the EPNs formulation for the successful use of EPNs as bio insecticides.

Isolation and characterization of Lepidoptera specific Bacillus thuringiensis strains predominantly from north-eastern states of India.

Both, the tobacco caterpillar Spodoptera litura (Fabricius) and the cotton bollworm Helicoverpa armigera (Hübner), are serious polyphagous pests causing considerable loss to crops. Indiscriminate use of chemical pesticides for controlling them has rather resulted in their resistance development. Microbial pesticides, Bacillus thuringiensis in particular, play an important role in pest management. Here, we isolated Bacillus thuringiensis-like bacteria from the soil samples primarily collected from North East region of India along with some states viz., Haryana, Punjab, Maharashtra, West Bengal and Uttarakhand and studied their toxicity against the above two insect pests at 10 µg/g along with standard strain B. thuringiensis subsp. kurstaki HD-1 and at 1 µg/g Pseudomonas fluorescens based MVPII expressing Cry1Ac toxin and AUG-5. Isolates AUG-5 and GTG-7 proved toxic to more than 75% larvae on the 4th as well as 7th day of the treatment of the neonates of H. armigera. The AUG-5 isolate was also effective against S. litura. Ten effective isolates (AUG-5, GTG-4, GTG-7, GTG-9, GTG-42, GTG-64, GTG-70, GTG-3S, GTG-4S and GTG-6S) were characterized using biochemical and 16S rDNA analysis. Nearly, all the isolates tested positive for utilizing monosaccharides. All selected B. thuringiensis isolates showed resistance to ampicillin and co-trimoxazole except AUG-5 to co-trimoxazole. AUG-5 and GTG-7 were highly toxic to both insects, and possessed cry1, cry1A and cry2 genes. These isolates AUG-5 and GTG-7 also contained high Cry1Ac (104.8 and 88.32 ng/mg) and Cry2Ab (3792 and 1305.9 ng/mg), respectively in their spore-crystal complex. Both, AUG-5 and GTG-7 isolates, could be considered for further development as bioinsecticides. The present study has established the diversity and richness of B. thuringiensis-like isolates in soils collected from north-eastern region of India.

Actividad insecticida y reguladora del crecimiento de extractos de blachnum chillones (blechnaceae) y condolía microphylla cav. (rhamnaceae), sobre larvas de gallería melonada (L.) (lepidoptera: pyralidae) / Insecticidal activity and growth regulatory of blechnum chilense (blechnaceae) and condalia microphylla cav. (rhamnaceae) extracts, on larvae of galleria mellonella (linnaeus) (lepidoptera: pyralidae)

Galleria mellonella (L.) es una de las plagas más importantes para la apicultura, debido a que en su estado larval se alimentan de la cera, polen y miel almacenados en los panales de Apis mellifera (L.). El objetivo del presente estudio fue determinar la actividad insecticida y reguladora del crecimiento de extractos obtenidos a partir de hojas y tallos de Blechnum chilense (Kaulf.) Mett y Condalia microphylla Cav. sobre larvas (L2) de G. mellonella. Los extractos fueron obtenidos con solventes orgánicos de diferente polaridad. Los resultados obtenidos indican que, el extracto de B. chilense obtenido con acetato de etilo y el extracto de C. microphylla obtenido con acetona, tienen efecto insecticida efectivo sobre larvas de G. mellonella y a aplicado en dosis sub-letales afectan negativamente la ganancia de peso larvario. A su vez, el extracto obtenido de B. chilense presentó actividad reguladora del desarrollo larvario de G. mellonella, induciendo el estado de pupa en forma prematura.

Galleria mellonella (L.) is the most detrimental pest to beekeeping, due the larvae feeds on hive of Apis mellifera (L.) consuming the wax, pollen and honey. The aim of this study was to determine the insecticidal activity and growth regulatory activity of extracts obtained from leaves and stems of Blechnum chilense (Kaulf.) Mett and Condalia microphylla Cav. for larvae of G. mellonella. The extracts were obtained with organic solvents of different polarity. The results show that the extract of B. chilense obtained with ethyl acetate, and the extract of C. microphylla obtained with acetone have effective insecticidal activity on larvae of G. mellonella, when applied at sub-lethal doses affect adversely the larval weight gain. In turn, the extract obtained from B. chilense showed regulatory activity on larval development of G. mellonella, inducing pupal stage prematurely.

The unfulfilled promises of scorpion insectotoxins

Since the description and biochemical characterization of the first insect-specific neurotoxins from scorpion venoms, almost all contributions have highlighted their potential application as leads for the development of potent bioinsecticides. Their practical use, however, has been hindered by different factors, some of which are intrinsically related to the toxins and other external determinants. Recent developments in the understanding of the action mechanisms of the scorpion insectotoxins and their bioactive surfaces, coupled with the exploration of novel bioinsecticide delivery systems have renewed the expectations that the scorpion insectotoxins could find their way into commercial applications in agriculture, as part of integrated pest control strategies. Herein, we review the current arsenal of available scorpion neurotoxins with a degree of specificity for insects, the progress made with alternative delivery methods, and the drawbacks that still preclude their practical use.(AU)

The unfulfilled promises of scorpion insectotoxins

Since the description and biochemical characterization of the first insect-specific neurotoxins from scorpion venoms, almost all contributions have highlighted their potential application as leads for the development of potent bioinsecticides. Their practical use, however, has been hindered by different factors, some of which are intrinsically related to the toxins and other external determinants. Recent developments in the understanding of the action mechanisms of the scorpion insectotoxins and their bioactive surfaces, coupled with the exploration of novel bioinsecticide delivery systems have renewed the expectations that the scorpion insectotoxins could find their way into commercial applications in agriculture, as part of integrated pest control strategies. Herein, we review the current arsenal of available scorpion neurotoxins with a degree of specificity for insects, the progress made with alternative delivery methods, and the drawbacks that still preclude their practical use.

Efficacy of Spodoptera litura multiple nucleopolyhedrovirus after serial passage through the homologous insect larval host.

An originally isolated baculovirus, Spodoptera litura multiple nucleopolyhedrovirus (SpltMNPV) was serially passed through the S. litura larvae for upto four generations to determine the mean number of occlusion bodies (OBs) harvested per larva and their efficacy in terms of infectivity, feeding cessation and speed of kill of host larvae. The results revealed that the mean number of OBs harvested per larva increased significantly with increase in the dose of SpltMNPV at each passage and the yield was significantly lower in original stock wild-type SpltMNPV (P0) as compared to serially passed SpltMNPV (P1, P2, P3 and P4). Laboratory bioassays indicate that median lethal doses (LD50), median times to feeding cessation (FT50) and median survival times (ST50) of P0, P1, P2, P3 and P4 were significantly different from each other. The OBs of each passage when tested for their cross-infectivity against Spodoptera exigua and Spilarctia obliqua revealed significant reduction in their mortality. These results indicate that serially passed SpltMNPV is more host specific and more effective biocontrol agent than the original stock wild-type virus and can be adopted for mass production as a viral pesticide for control of the S. litura.

Citrullus Colocynthis (Linn.) and Luffa Acutangula (L.) Roxb, Schrad. Source of Bioinsecticides and their Contribution in Managing Climate Change.

In present agricultural practices greenhouse gases are created in a multitude of ways. Pesticide and fertilizer applications, irrigation, lighting, transportation, and other machinery are powered by greenhouse gas-emitting fossil fuels. The fertilizers and pesticides alone are one of the largest contributors to greenhouse gas emissions in agriculture. The overuse of agricultural chemicals pollutes watersheds and kills plants that could otherwise capture greenhouse gases and actually reduce global warming. Compared to conventional agriculture, organic agriculture is reported to be more efficient and effective both in reducing GHGs (CO2, CH4 and N2O) emission mainly due to the less use of chemical fertilizers and pesticides. Organic agriculture also reported to be climate change resilience farming systems as it promotes the proper management of soil, water, biodiversity and local knowledge there by acting as a good options for adaptation to climate change. In the present study two of the cucurbits are studied for their insecticidal acitivity against Aphids one is a wild specie Citrullus colocynthis (Linn.) Schrad.and another one is cultivated specie that is Luffa acutangula (L.) Roxb., both have found effective but C. colocynthis found more effective than L. Acutangula.