Lethal and trans-generational effects of Metarhizium anisopliae on red palm weevil,Rhynchophorus ferrugineus,using age-stage,two-sex life table

Outbreaks of red palm weevil, Rhynchophorus ferrugineus Olivier, in Pakistan are considered a very serious threat to the date industry, owing to the sudden death of date palm trees. Current integrated management strategy largely depends on insecticides, with negative impacts on the environment. Integration of bio-control strategies incorporating entomopathogenic fungi (EPF) may provide an ecologically safe alternative. In the current study the effectiveness of Metarhizium anisopliae Sorokin against R. ferrugineus was investigated using the immersion method and the effects of this fungus on biological parameters of R. ferrugineus were estimated by age-stage, two-sex life table software. After treatment with M. anisopliae isolate (Ma-M₂), lethal concentration (LC₅₀) 1.2 × 10⁶ spores/mL was determined against R. ferrugineus 3rd instar grubs. M. anisopliae infected grubs showed reduced net reproductive rate R_o (75.55) and mean generation time T (113.886 days) and intrinsic rate of increase r (0.03797) compared to the untreated group. Also, adult longevity of males and females were significantly reduced in the M. anisopliae-infected group (F₁) as compared to the untreated group. Trans-generational studies revealed that grubs stage duration (69.34 days) and fecundity (181.32 eggs) of the M. anisopliae-infected group (F₁) were decreased compared to the untreated group. The use of lasting and adverse effects on the biological parameters of R. ferrugineus, indicate that treatment with EPF may provide an eco-friendly management strategy for this pest.     

Studies on host preference and its biological effects on the red palm weevil,Rhynchophorus ferrugineus Olivier in Egypt The fruit trees

A comparative bioassay (choice or non-choice) was developed to evaluate the efficacy of different species of fruit trees as alternative host plants on the morphological and biological aspects of the red palm weevil, Rhynchophorus ferrugineus and also the latent effect of feeding the larvae on pupal and adult stages. Highly significant differences exist between averages of larval body weight of R. ferrugineus larvae fed on fruit trees. The lowest average of larval weight occurred in the larvae fed on the pieces of sour orange, mandarin followed by olive and lime trees. A small increase in body weight appeared in larvae fed on mango, fig and guava in comparison with those fed on date palm trees (control). The larval mortality reached the highest percentages (100%) by obligatory feeding of R. ferrugineus insects on olive, lime, sour orange and mandarin followed by mango (97%) and guava (96%) compared with control (0%), respectively. The latent effect of obligatory feeding on the larvae of R. ferrugineus was highly decreased during pupation (4%, 4% and 8%) when the insect fed on mango, guava and fig, compared with 100% pupation in the case of larvae fed on ornamental palm or date palm, respectively. Most pupae resulting from the treated larvae by R. ferrugineus failed in adult emergence especially those insects fed on olive, lime, sour orange and mandarin trees compared to untreated insects (95.6–96.2%), respectively. Data also proved that if an insect is forced to feed on a non-preferred host plant, this results in death of the insect often during the larval stage, a decrease of pupation or failure of adult emergence. These results led to the conclusion that mango, fig, and guava trees are believed to be vulnerable hosts for red weevil attack. The lowest percentage of food consumption appeared in those larvae fed on mandarin, sour orange, lime and olive trees compared with those fed on palm. The present results emphasise the need to test the efficiency of the compounds extracted from these host plants if it is considered as an insecticide or an alternative host protectant for management of the red palm weevil R. ferrugineus.     

Parasites rather than phoronts: Teratorhabditis synpapillata nematodes reduce lifespan of their Rhynchophorus ferrugineus host in a life stage‐dependent manner

Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) red palm weevils are often reported in association with different organisms including nematodes. The significance of this interaction and whether nematodes can influence their life‐history traits is unclear. We collected Rhynchophorus ferrugineus red palm weevils at different developmental stages and locations in Tunisia, observed and dissected them in search for nematodes and other interacting organisms, established laboratory colonies and identified the nematodes associated with them, and conducted nematode–insect interaction assays to determine the capacity of the nematodes to influence their life‐history traits. We observed Beauveria bassiana fungi in larvae, nymph, and adults; Centrouropoda and Uroobovella acari associated with the adults, and Teratorhabditis synpapillata nematodes associated with larvae and adults. Nematode–insect interaction bioassays revealed that T. synpapillata nematodes reduce the lifespan of the insect larvae in a population‐dependent manner, but do not influence the lifespan of adults. Our study uncovers an important factor that may determine population dynamics of this important palm pests and provides evidence to conclude that these organisms establish a parasitic relationship, rather than a phoretic relationship.     

Assessing Spirulina platensis as a dietary supplement and for toxicity to Rhynchophorus ferrugineus (Coleoptera: Dryopthoridae)

Insects are important for humanity; play role in crop pollination, and biocontrol of harmful pests. The red palm weevil, Rhynchophorus ferrugineus, is a major pest of date palms and has become a serious threat. Scientists needs ample numbers of insects for bioassays to explore control options. The alga Spirulina platensis, is enriched by protein, natural vitamins, minerals, and amino acids, stimulate the development of organisms that feed on it. I assessed the value of Spirulina as a nutritional supplement for red palm weevil larvae by adding its various percentages to the artificial diet. Once a week, the larvae were removed from the containers, washed with distilled water, dried, weighed using an electronic scale, returned to a new container, and supplied with Spirulina mixed fresh diet. Larvae fed with lower concentrations showed vigorous growth and significant weight gain. Particularly, larvae fed 0.5%, 1%, and 2% Spirulina powder supplementation to their diet were healthier and gained more weight than larvae reared with >5% concentration. Overall 40% mortality was recorded in larvae fed with 10% concentration. Higher concentrations were lethal, and all larvae died within two weeks when fed 20% Spirulina. The present research findings indicate that Spirulina used in concentrations from 0.5% to below 5% had a beneficial effect on red palm weevil larval growth but a detrimental effect and even mortality was recorded when used ≥5%.     

Potential of an indigenous strain of the entomopathogenic fungus Beauveria bassiana as a biological control agent against the Red Palm Weevil, Rhynchophorus ferrugineus

The potential of a strain of Beauveria bassiana (Ascomycota: Clavicipitaceae) obtained from a naturally infected Rhynchophorus ferrugineus (Coleoptera: Curculionidae) pupa as a biological control agent against this weevil was evaluated both in the laboratory and in semi-field assays. Laboratory results indicate that this strain of B. bassiana can infect eggs, larvae and adults of R. ferrugineus (LC₅₀ from 6.3 × 10⁷ to 3.0 × 10⁹ conidia per ml). However, mortality was not the only indicator of treatment efficacy because adults of either sex inoculated with the fungus efficiently transmitted the disease to untreated adults of the opposite sex, with male-to-female and female-to-male rates of transmission of 55% and 60%, respectively. In addition, treatment with B. bassiana significantly reduced fecundity (up to 62.6%) and egg hatching (32.8%) in pairing combinations with fungus-challenged males, females or both sexes. Likewise, 30-35% increase in larval mortality was observed in larvae obtained from eggs from fungus-challenged females or from untreated females coupled with inoculated males, resulting in an overall 78% progeny reduction. Semi-field preventive assays on potted 5-year old Phoenix canariensis palms, with efficacies up to 85.7%, confirmed the potential of this strain as a biological control agent against R. ferrugineus.     

Biological activity of Pakistani isolate SpltNPV-Pak-BNG in second,third and fourth instar larvae of the leafworm Spodoptera litura

Recently, a novel isolate of the baculovirus Spodoptera litura nucleopolyhedrovirus (SpltNPV) has been isolated from Pakistan, which is distinct from the type species SpltNPV-G2 (ICTV). Here, we examined the biological activity of this isolate (SpltNPV-Pak-BNG) in second (L2), third (L3) and fourth instar (L4) larvae of the leafworm S. litura, more specifically to measure biological properties that are relevant for use of this virus for pest control under field conditions. The median lethal dose for L2 and L3 instar larvae was similar, but significantly lower than for L4 larvae. Likewise, the survival time was similar for L2 and L3 larvae (84?h), but was significantly longer for L4 instar larvae (108?h). Thus, in terms of efficacy, S. litura L2 and L3 instar larvae are the preferred targets for S. litura control with SpltNPV-Pak-BNG in field crops in Pakistan. On the basis of our data spray regimes can be designed to control the leafworm in cotton and vegetable crops targeting L2 and L3 larvae.     

Use of Beauveria bassiana in combination with commercial insecticides to manage Phauda flammans (Walker) (Lepidoptera: Phaudidae): Testing for compatibility and synergy

An entomopathogenic fungal strain, Beauveria bassiana PfBb, was identified from Phauda flammans (Lepidoptera: Phaudidae) larvae. The compatibility and synergy of B. bassiana PfBb employed in combination with three concentrations (i.e., recommended concentration, 20% and 10% of the recommended concentration) of five commercial insecticides were determined. Beta cypermethrin at 10% of recommended concentration had the lowest inhibitory effect on the mycelial growth of B. bassiana PfBb compared with other insecticides. Insecticides utilized at recommended concentration had no significant effect on the sporulation of B. bassiana PfBb, while the extent of their effect at 20% and 10% of recommended concentration differed among insecticides. Insecticides at 10% of recommended concentration had the lowest inhibition of sporulation and conidial germination compared with other concentrations. The conidial germination of B. bassiana PfBb was the highest after treatment with beta cypermethrin at 10% of recommended concentration. The cumulative mortality for 1 × 10⁷ spores/mL B. bassiana PfBb combined with each insecticide at 10% of recommended concentration was higher than that observed with the application of insecticides alone. The percent cadavers of Phauda flammans larvae observed after treatment with B. bassiana PfBb combined with beta cypermethrin at 10% of recommended concentration were not significantly different from those observed after infection with B. bassiana PfBb alone. Our findings demonstrate that B. bassiana PfBb combined with beta cypermethrin at 10% of recommended concentration could increase the efficiency of this insecticide.     

Susceptibility of preimaginal western cherry fruit fly, Rhagoletis indifferens (Diptera: Tephritidae) to Beauveria bassiana (Balsamo) Vuillemin Clavicipitaceae (Hypocreales)

Last-instar larvae of the western cherry fruit fly, Rhagoletis indifferens, were subjected to Beauveria bassiana GHA incorporated into sterile sand and non-sterile orchard soil. Mycosis in the pupal stage was observed in >20% of buried R. indifferens pupae and >80% of larvae entering sand treated with either of two B. bassiana isolates. When pre-pupal larvae burrowed into conidium-treated non-sterile cherry orchard soil, the incidence of mycosis, on both the puparia and internally developing pupae, increased with dose. Internal pupal tissues were found to contain B. bassiana. Increasing the soil moisture level from 20% to 35% water holding capacity did not have an effect on the percentage of mycosed pupae. This is the first evidence that the preimaginal stages of R. indifferens are susceptible to infection by B. bassiana.     

Entomopathogenic fungi-mediated biological control of the red palm weevil Rhynchophorus ferrugineus

The red palm weevil (RPW), Rhynchophorus ferrugineus, is an important pest of palms, and difficult to control by conventional methods. Therefore, microbial control is an alternative strategy for controlling RPW. Herein, a total of 15 entomopathogenic fungi (EPFs) were subjected to primary pathogenicity screening against last stage of RPW larvae. The preliminary data showed that four Beauveria bassiana isolates (JEF-484, 158, 462 and 507) and one Isaria fumosorosea isolate (JEF-014) resulted in 100 % mortality within 5–10 days post inoculation (d.p.i.), respectively. According to the time required for RPW mortality, JEF-484, 158, 462 and 014 were further subjected to bioassays using 10⁷ conidia/ml suspensions by spraying method. Based on the results, JEF-484 showed the highest mortality and shortest LT₅₀ on the last stage of RPW larvae, followed by JEF-158. The two isolates also showed good conidial production and high thermal stability compared to the other isolates. Therefore, JEF-484 and JEF-158 were selected for bioassays against RPW egg and the last larval stage with different concentrations of 10⁵, 10⁶ and 10⁷ conidia/ml conidial suspensions by spraying method. For the bioassay at the egg stage, JEF-158 showed a significantly higher ovicidal effect than JEF-484. In the larval bioassay, both EPF isolates showed a dosage-dependent effect on the RPW larvae. JEF-484 caused higher mortality in RPW larvae than JEF-158. In summary, the combination of the 2 promising EPF isolates might provide an opportunity for the practical microbial control of RPW at different life stages in palm tree fields.     

Hemocytes of Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) and their response to Saccharomyces cerevisiae and Bacillus thuringiensis

Originally from tropical Asia, the Red Palm Weevil (RPW Rhynchophorus ferrugineus (Olivier)) is the most dangerous and deadly pest of many palm trees, and there have been reports of its recent detection in France, Greece and Italy. At present, emphasis is on the development of integrated pest management based on biological control rather than on chemical insecticides, however the success of both systems is often insufficient. In this regard, RPW appears to be one pest that is very difficult to control. Thus investigations into the natural defences of this curculionid are advisable. RPW hemocytes, the main immunocompetent cells in the insect, are described for the first time. We identified five hemocyte cell types from the hemolymph of R. ferrugineus: plasmatocytes (∼50%), granulocytes (∼35%), prohemocytes (∼8%), oenocytes (∼4%) and spherulocytes (∼3%). SEM observations were also carried out. Some aspects of RPW interaction with non-self organisms, such as Saccharomyces cerevisiae and the entomopathogen bacterium, Bacillus thuringiensis (Bt), are discussed. Plasmatocytes and granulocytes were involved in nodules and capsule formation as well as in the phagocytosis of yeast. The hemocyte response of RPW larvae to sub-lethal doses of commercial products containing Bt was examined. In vivo assays were carried out and Bt in vegetative form was found in the hemolymph. After a diet containing Bt, the number of total hemocytes, mainly plasmatocytes, in the RPW larva hemolymph declined sharply (∼12%) and then remained at a low level, while the number of other circulating cells was almost unchanged.     

Nano essential oils against the red palm weevil,Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae)

The red palm weevil, Rhynchophorus ferrugineus, is the biggest pest of various palm species. The aim of this study was to evaluate the impact of the essential oils of purslane, mustard and castor (bulk and nano) on R. ferrugineus and to evaluate the oviposition deterrent effect to prevent new infestations. Prepared concentrations of essential oils (bulk and nano) were tested for their impact on R. ferrugineus larvae, pupae and adults. The most effective oil was purslane oil (bulk and nano), followed by mustard and the least effective was castor. The percent mortality of larvae was 75.2, 45.3 and 17.9% (bulk phase) and 92.5, 84.4 and 65.5% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The percent mortality of adults was 67.2, 40.0 and 15.6% (bulk phase) and 83.5, 72.9 and 60.0% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The number of eggs laid decreased with increasing concentration of the tested oils. Purslane essential oil showed good impact on R. ferrugineus larvae and adults followed by mustard and castor oils as bulk and as nano.     

Pathogenicity of Beauveria bassiana isolated from Moroccan Argan forests soil against larvae of Ceratitis capitata (Diptera: Tephritidae) in laboratory conditions

The Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae), is the major tephritid pest in Morocco. This pest survives in Moroccan forests Argania spinosa and continually invades the nearest agricultural areas. Entomopathogenic fungi are an interesting tool for fruit fly control and hold a useful alternative to conventional insecticides. However, primary selection of effective pathogens should be taken in laboratory condition prior to applying them in the field. Here, we used third late instar larvae of C. capitata to investigate the effectiveness of 15 local Beauveria bassiana isolates. Results showed that all isolates were able to infect the larval stage, producing a large mortality rate in puparia ranging from 65 to 95 % and caused significant reduction in adult emergence. The fungal treatments revealed that the mycosis occurred also in adults escaping infection as pupariating larvae. The percentage of mycosed puparia was highest in strain TAM6.2 (95 %) followed by ERS4.16 (90 %), therefore they were the most virulent. Median lethal concentration (LC₅₀) was studied for five isolates at four concentrations ranging from 10⁵ to 10⁸ conidia ml^?1. The results showed that the slopes of regression lines for B. bassiana ERS4.16 (slope = 0.386) and TAM6.2 (slope = 0.41) were the most important and had the lowest LC₅₀ values (2.85 × 10³ and 3.16 × 10³ conidia ml^?1 respectively). This investigation suggests that the soil of Argan forests contains pathogenic B. bassiana isolates and highlights for the first time their potential as biological control toward C. capitata larval stage in Morocco.     

Purification and expression of glutathione S-transferase from a Malaysian population of Aedes albopictus (Diptera: Culicidae)

Glutathione S-transferase (GST) from the 4^th instar larvae of the dengue vector Aedes albopictus was purified by glutathione-agarose affinity chromatography and characterised using SDS-PAGE. The expression of the purified enzyme in the life stages and insecticide treated populations of Ae. albopictus as well as its cross-reactivity with larval GST of two dipteran species Aedes aegypti and Batrocera papayae were observed using western blotting. The purified GST had a specific activity of 196.0 ± 11 μmol/min/mg with a purification fold and yield of 28 and 69%, respectively. The SDS-PAGE analysis of the purified GST depicted a single band size of 23 kDa. The GST was expressed in all the larval and adult stages of Ae. albopictus with the exception of the pupal stage. However, the expression level in the adult stage was visibly reduced as compared to the larval stages. Western blotting analysis showed no cross-reactivity with the GST of Ae. aegypti (4^th instar) and B. papayae (3^rd instar) larvae. The expression of this enzyme was not inducible by exposure to the insecticides dichlorodiphenyltrichloroethane (1.25 mg/L) and malathion (0.3125 mg/L).     

Impact of the invasive plant species “Nicotiana glauca” toxins on the larvae of the invasive insect species “Rhynchophorus ferrugineus”: A damaging pest of date palm trees in Saudi Arabia

The wild tree tobacco (Nicotiana glauca) is an alien species that invaded vast areas of the Southwestern region of Saudi Arabia. While, the Red Palm Weevil (RPW) (Rhynchophorus ferrugineus) is considered to be the most damaging invasive insect species of palm trees all over the kingdom of Saudi Arabia, causing major economic losses to farmers and the economy of the country. Using conventional insecticides to control harmful insects such as RPW has undesirable effects on the environment and human health. Alternatively, using biocontrol agents such as poisonous extracts from N. glauca might be a better approach in pest management and can be considered as an eco-friendly, cost-effective, and safe alternative. Therefore, the current study aimed to evaluate the larvicidal effect of N. glauca aqueous extracts against the red palm weevil larvae. The plant specimens were collected from Al-Baha region in the Southwest of Saudi Arabia. Each single test consisted of 20 larvae, and N. glauca preparations were; 1, 1.5, 2, 2.5, and 3 ml, besides the control test. Results obtained for the effect of botanical extracts; leaf, flower, stem and root against R. ferrugineus larvae for an exposure period of 24 hr. at the concentrations of 2.8, 4.2, 6.0, 7.0 and 8.0 ppm. The concentrations for N. glauca extracts reflected an LC₅₀ of 2.7 ppm for leave, 2.6 ppm for flower, 2.8 ppm for stem and 7.00 ppm for root. While, the same concentrations extracts reflected an LC₉₅of 11 ppm for leaf, 9.6 ppm for flower, 8.9 ppm for stem and 13.00 ppm for root. These results showed that N. glauca extracts have a remarkable potentiality as insecticidal substances that can be used as an ecofriendly integrated approach for the management of R. ferrugineus.     

Attraction of entomopathogenic nematodes Steinernema carpocapsae and Heterorhabditis bacteriophora to the red palm weevil (Rhynchophorus ferrugineus)

The red palm weevil (RPW), Rhynchophorus ferrugineus, is a serious pest of date palms. Its larvae bore deep into the trunk disrupt the vascular tissues and kill the infested trees. Behavioral features of entomopathogenic nematodes (EPNs), reflected by attraction and distribution patterns, are fundamental aspect in determining their parasitic ability and potential management of RPW. We studied the attraction behavior of the EPNs Steinernema carpocapsae and Heterorhabditis bacteriophora to the RPW under simulated natural conditions in tubes to evaluate their infective potential. In all experiments, a certain proportion of infective juveniles (IJs) (16–20%) stayed near the inoculated site and a major proportion (38–48%) was attracted to the host end. Both H. bacteriophora and S. carpocapsae were efficient crawlers, climbing up and descending when locating their insect host. They were efficiently attracted to the various larval sizes and stages of the RPW life cycle. Host localization by ascending movement was more prominent in S. carpocapsae than in H. bacteriophora. In general, H. bacteriophora is classified as a cruiser forager and S. carpocapsae as an ambusher. However, in this study, we discovered a higher percentage of cruiser foragers among S. carpocapsae IJs. They dispersed much faster and their cruising behavior was prominent characteristic in controlling the cryptic RPW concealed in organic habitats.     

Virulence of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against red palm weevil,Rhynchophorus ferrugineus (Olivier)

Nineteen different isolates of the entomopathogenic fungi Beauveria bassiana sensu lato (s.l.) and Metarhizium anisopliae s.l. (Ascomycota: Hypocreales), recovered from different soil samples (field crops, fruit orchards, vegetable fields and forests) and insect cadavers were tested against Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) at two different spore concentrations (1 × 10⁷ and 1 × 10⁸ conidia mL^?1). Three isolates of B. bassiana and two of M. anisopliae gave >88 % larval and >75 % adult mortality of R. ferrugineus on their highest dose rate respectively. More sporulating cadavers (mycosis) resulted from a high dose rate compared to low dose on both life stages of R. ferrugineus. The current study confirmed the lethal action of B. bassiana and M. anisopliae isolates with mortality levels usually directly proportional to the conidial concentration. This study further confirmed that the isolates recovered from R. ferrugineus dead cadavers gave more mortality compared to the other sources. In the virulence assay two isolates of B. bassiana caused the highest percentage of both larval and adult mortality at all exposure intervals which suggest that they may be the most promising for use in sustainable management programs aimed at microbial control in date palm orchards.     

Toxicity and Detoxification Mechanism of Black Pepper and Its Major Constituent in Controlling <Emphasis Type="Italic">Rhynchophorus ferrugineus</Emphasis> Olivier (Curculionidae: Coleoptera)

The survival, feeding response, and detoxification mechanism of Rhynchophorus ferrugineus Olivier, a key pest responsible for destruction of date palm, was examined with different extracts of Piper nigrum and its major constituent (piperine) identified by GC-MS. In the present study, toxicity of different extracts of black pepper was evaluated by incorporating diffferent doses of extracts into the artificial diet of red palm weevil larvae. All extracts showed dose-dependent insecticidal activity to the tested eighth-instar red palm weevil larvae. Among all the extracts, maximum larvicidal activity was exhibited by chloroform (LD₅₀ = 342.62 mg/l), followed by dichloromethane (LD₅₀ = 357.78 mg/l), acetone (LD₅₀ = 372.57 mg/l), and ethanol (LD₅₀ = 408.88 mg/l). However, piperine, a major constituent of all black pepper extracts identified by GC-MS in the present work, was found to be the most potent treatment exhibiting the least LD₅₀ (219.88 mg/l). In addition, nutritional indices evaluated by calculating the efficiency of the conversion of ingested food (ECI) and digested food (ECD) at the same dose (219.88 mg/l) showed that there was maximum reduction in the ECI (49.90%) and ECD (62.21%) index of larvae fed diets incorporated with piperine. Larvae that were fed diets incorporated with different black pepper extracts experienced increases in the expression of detoxification genes (glutathione S-transferase and cytochrome P450), and this upregulation in detoxification genes (glutathione S-transferase, cytochrome P450 and esterase) was tremendously high in larvae fed diets incorporated with piperine. Results suggest that piperine is a promising bio-pesticide agent for the control of R. ferrugineus Olivier.     

Toxicity of spinosad and Beauveria bassiana to the black cutworm,and the additivity of subletal doses

The main objective of this study was to assess the susceptibility of the black cutworm (Agrotis ipsilon) to the biopesticide spinosad and to a commercial formulation (GHA strain) of the entomopathogenic fungus, Beauveria bassiana. Secondly, we quantified the effects of sublethal doses of spinosad on a number of A. ipsilon fitness parameters, and interactions resulting from simultaneous applications of sub-lethal doses of spinosad and B. bassiana. Under laboratory conditions, A. ipsilon third instar larvae were highly susceptible to spinosad, with an estimated LC₅₀ of 50 ppm. The entomopathogenic fungus, B. bassiana had a lower efficacy with an estimated LC₅₀ of 7×107 spores mL^?1. Topical applications of 5, 7.5 and 10 ppm of spinosad on third instar larvae reduced larval size and increased time to pupation and to emergence. However, pupal and adult weights were not significantly different between treated and control individuals. Additivity was observed from most spinosad–B. bassiana combinations tested, thus indicating compatibility between products. We concluded that spinosad is a promising tool for controlling black cutworm larvae alone or in combination with other products.     

Efficacy of some biorational insecticides against Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) under laboratory and greenhouse conditions in Kuwait

The tomato leafminer, Tuta absoluta, is one of the most destructive pests of tomato worldwide. Management of the pest is mainly based on chemical insecticides. Reliance on insecticides is difficult to sustain because of unintended long‐term adverse effects on the environment and human health. Consequently, there is a need to develop pest management strategies that ensure the production of high‐quality products, while at the same time ensuring environmental sustainability and maximum consumer protection. We evaluated the efficacy the biopesticides: Azadirachtin, Bacillus thuringiensis, Steinernema feltiae and Beauveria bassiana individually and in combination against T. absoluta under laboratory and greenhouse conditions. When second instar larvae were exposed to tomato leaf discs treated with Azadirachtin (3 g / L), B. thuringiensis (0.5 g/L) or B. bassiana (1.5 g/L), 70%–86%, 55%–65%, and 45.5%–58.5% mortality was observed, respectively. Steinernema feltiae (1,000 IJs/ml) was the least effective biopesticide, with 26%–42% mortality. In the greenhouse trials on tomato, pest infestation (mines/10 leaves/plant) and fruits damaged were significantly lower on plants treated with Azadirachtin  +  B. thuringiensis or Azadirachtin  +  B. bassiana compared to plants treated with Azadirachtin, B. thuringiensi, B. bassiana or S. feltiae alone. Azadirachtin  +  B. thuringiensis and Azadirachtin  +  B. bassiana resulted in 90% and 81% reduction in fruits damaged in the summer experiments, respectively, and 96% and 91% in winter. The most severe pest infestation was observed on plants treated with S. feltia. The results indicate that the biopesticides, except S. feltia, can contribute to T. absoluta control in greenhouse tomato crops. In particular, the combined use of Azadirachtin with B. thuringiensis or B. bassiana provided the highest level of control of the pest. The potential for including these biopesticides in an overall sustainable integrated pest management programme for T. absoluta is discussed.     

Potential of a combination of entomopathogenic fungal strains and a non-ionic surfactant to control the fall armyworm (Spodoptera frugiperda)

The fall armyworm, Spodoptera frugiperda, is an emerging invasive pest in Taiwan that feeds on a wide range of crops and causes serious damage. Herein, an entomopathogenic fungal library (EFLib) was constructed to identify potential microbes to control FAW. Twenty-eight indigenous entomopathogenic fungi (EPF) were isolated and investigated for their potential pathogenicity, with Metarhizium pinghaense (Mp-NCHU-124) and Beauveria bassiana (Bb-NCHU-157) exerting dose-dependent effects on the 4th instar FAW larvae. The non-ionic surfactant Silwet L-77 rapidly killed FAW larvae after spraying at a concentration of 300 mg/kg and the toxic effect of Silwet L-77 on FAW larvae was dose-dependent. When the EPF isolates (10⁶ conidia/mL) were applied to FAW larvae in combination with the non-ionic surfactant Silwet L-77 (30–90 mg/kg), the mortality rate dramatically increased and the LT₅₀ reduced, with increased fungal mycosis (Mp-NCHU-124: 38% to 72% and Bb-NCHU-157: 20 to 62%), indicating the high compatibility of EPF with the non-ionic surfactant. Thus, the Silwet L-77+EPF combined formulation has potential for practical field application for FAW pest control and sustainable agriculture in the future.