The side-effects of pesticides used in integrated production of peaches in Brazil on the egg parasitoid Trichogramma cacoeciae Marchal (Hym., Trichogrammatidae)

Abstract: The side‐effects of six pesticides used on peaches in Brazil were tested on the hymenopteran egg parasitoid Trichogramma cacoeciae using four laboratory tests: (a) adult parasitoid exposure to fresh pesticide residue on glass plates (worse case); (b) direct spray of host eggs enclosing the parasitoid egg, larvae or pupae (less‐exposed life stages); (c) exposure of adults to pesticide residues on plant leaves at different intervals after application (persistence); (d) Dose–response pesticide exposures of adults on glass plates. Two dose rates were used: (1) The highest recommended field dosage (FD) and (2) the predicted initial environmental concentrations (PIEC). The results showed that the preparations greatly differed in their initial toxicity and persistence. The insecticide Valient^® (methoxyphenozide) and the fungicide Venturol^® (dodine) were considered harmless to T. cacoeciae adults as they fell into the class 1 category according to the guidelines of the International Organization for Biological Control (IOBC) when parasitoids were directly exposed to chemical residues. The insecticide/acaricide Assist^® (mineral oil) was slightly harmful at the rate of PIEC 0.4 (40% of FD) and moderately harmful at FD. Pesticides in the categories harmless and moderately harmful can be considered for use in integrated pest management (IPM). The fungicide/acaricide Kumulus DF^® (sulphur) and the insecticides Dipterex^® 500 (triclorfon) and Lebaycid^® 500 (fenthion) were harmful at both concentrations. In the persistence test, Assist^® was short lived and therefore may in special cases (i.e. reduced direct contact) be considered for use in IPM, but Kumulus^® DF, Dipterex^® 500 and Lebaycid^® 500 constantly reduced parasitism between 77 and 100% and were rated as persistent (more than 30 days). The direct spray of parasitized host eggs at intervals after parasitism showed that Assist^® and Kumulus^® DF were harmless to the parasitoid egg, larvae and pupae within the host eggs. Dipterex^® 500 was slightly harmful when sprayed one day after parasitism (parasitoid egg) and moderately harmful to the other two stages (larvae and pupae). Lebaycid^® 500 was harmful to the parasitoid egg and larvae and moderately harmful to the pupae. The dose–response test showed that Kumulus^® DF and Dipterex^® 500 were toxic to T. cacoeciae. Kumulus^® DF was harmful from 1 PIEC 0.4 to 0.125 PIEC 0.4 dosages and was slightly harmful with 0.0625 PIEC 0.4 dosage. Dipterex^® 500 was harmful to T. cacoeciae in all the dosages tested.     

Growth ofDiacrisia obliqua [Lep.: Arctiidae] with low doses ofBacillus thuringiensis var.Kurstaki

Early 3rd instarDiacrisia obliqua Walk. larvae were treated with concentrations ofBacillus thuringiensis var.kurstaki (Dipel^®) and the growth of treated larvae was assessed. All the doses reduced significantly the weight and survival of the insects (p<0.001).     

Assessing risks of pesticides targeting lepidopteran pests in cruciferous ecosystems to eggs parasitoid,Trichogramma brassicae (Bezdenko)

Lethal and sub lethal effects of fresh and old residues of azadirachtin, spinosad, Bacillus thuringiensis var. kurstaki (Bt var. k), and deltamethrin, were evaluated at their recommended field doses against adult and immature stages of Trichogramma brassicae under in vitro conditions. The experiments were carried out at the Entomology section of Division of Crop Protection, ICAR Research Complex for NEH region, Umiam, Meghalaya, in 2012–2013. The effects of different pesticides were determined by bioassays using the residual film method, the diet contamination method, the pupal dip method and the topical application technique. The four pesticides were found harmful to adult T. brassicae after ingestion, however surface contact bioassays revealed that Bt var. k was the least toxic pesticide. Except Bt var. k, other three pesticides were found harmful also to the immature stages of T. brassicae and significantly affected parasitism potential, adult emergence, longevity of adults, and sex ratio of the progeny. Deltamethrin and azadirachtin were the most harmful, even after 15 days of application. Spinosad was found to be relatively safe to T. brassicae after 15 days of application. As Bt appeared to be the least toxic pesticide for T. brassicae, it could be used for the management of severe infestations of lepidopteran pests in cruciferous ecosystems.If essential, spinosad may be used 15 days after parasitoid release, thus minimizing the chances of parasitoid exposure.     

Pesticide selectivity for the insect egg parasitoid <Emphasis Type="Italic">Telenomus remus</Emphasis>

We evaluated the side-effects of insecticides, herbicides and fungicides on adults of the egg parasitoid Telenomus remus (Nixon) under laboratory conditions. The protocol was adapted from that proposed by the Pesticides and Beneficial Organisms Working Group of the International Organization for Biological Control (IOBC) for Trichogramma cacoeciae (Marchal). Chlorpyrifos, acephate, beta-cyfluthrin + imidacloprid, spinosad, and pyrethroids were harmful to the parasitoid, whereas methoxyfenozide, diflubenzuron, and flufenoxuron had no effect. Of the herbicides examined, only glyphosate + imazethapyr and 2,4-D amine were classified as harmless on the first and second days of parasitism; paraquat was the most harmful. Other herbicides were harmless on the first day of parasitism, but caused various levels of reduction of T. remus parasitism on the second day. The fungicides were harmless or only slightly harmful.     

Low Temperature Storage of <Emphasis Type="Italic">Telenomus remus</Emphasis> (Nixon) (Hymenoptera: Platygastridae) and its Factitious Host <Emphasis Type="Italic">Corcyra cephalonica</Emphasis> (Stainton) (Lepidoptera: Pyralidae)

We conducted three bioassays to evaluate the effect of low-temperature storage of eggs (host) and pupae and adults (parasitoid) on the biology and parasitism capacity of the egg parasitoid Telenomus remus (Nixon) (Hymenoptera: Platygastridae). Viable stored Corcyra cephalonica (Stainton) (Lepidoptera: Pyralidae) eggs were parasitized to the same degree or even higher than fresh eggs when stored until 14 days at 5°C or until 21 days at 10°C. In contrast, the percentage of parasitized sterilized eggs was equal to the control only when stored for 7 and 14 days. Survival of T. remus pupae declined with storage time at both studied temperatures (5 and 10°C). However, after 7 days of storage, survival of pupae was still 86.3 and 64.9% at 10 and 5°C, respectively. The number of adult male survivors remained similar until the fourth storage day at both 5 and 10°C. In contrast, female survival did not differ until day 8 at 10°C or day 6 at 5°C. Parasitism capacity of stored adults was not altered by storage compared with the control. Therefore, we conclude that the maximal storage time at 10°C is 21 days for viable C. cephalonica eggs and 7 days for T. remus pupae, while parasitoid adults should not be stored for more than 4 days at either 5 or 10°C.     

Does Bacillus thuringiensis have adverse effects on the host egg location by parasitoid wasps?

This study investigated the interaction between two pest biological control agents, the parasitoid wasp Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) and the entomopathogen Bacillus thuringiensis (Bacillales: Bacillacea) (Bt). The aim of this study was to evaluate if the presence of Bt (formulated products Agree^®, Dipel^® and HD1 and HD11 strains) interferes in the oviposition preference of T. pretiosum to eggs of Helicoverpa zea (Lepidoptera: Noctuidae). Using an olfactometry test, the eggs of H. zea were bathed with the commercial formulations, with the Bt suspensions or distilled water, and offered to the parasitoid wasps in order to evaluate parasitism. The results showed that H. zea eggs sprayed with commercial formulations and Bt strains did not interfere in the choice made by the parasitoid. The parasitoid wasp is not able to distinguish between eggs with or without B. thuringiensis treatment, independently of strains suspension or commercial formulations. Therefore, these two control agents may be used together without negative interaction.     

Efficacy of entomopathogenic bacteria for control of Musca domestica

The aim of this study was to evaluate the larvicidal activity, and sub lethal effects of entomopathogenic bacteria Brevibacillus laterosporus, Bacillus thuringiensis var. israelensis, B. thuringiensis var. kurstaki, and a commercial formulation of Bacillus sphaericus on Musca domestica. Bacterial suspensions were prepared in different concentrations and added to the diet of newly-hatched larvae which were monitored until the adult stage. The larvae were susceptible to the B. laterosporus, B. thuringiensis var. israelensis, and B. thuringiensis var. kurstaki bacteria in varied concentration levels. These bacteria have larvicidal and sub lethal effects on the development of flies, reducing both adult size, and impairing the reproductive performance of the species.     

Extrachromosomal DNA in Bacillus thuringiensis var. kurstaki, var. finitimus, var. sotto, and in Bacillus popilliae

Four entomopathogenic bacteria contained extrachromosomal deoxyribonucleic acid (DNA) molecules of various sizes. Bacillus thuringiensis var. kurstaki contained twelve elements banding on agarose gels that ranged from 0.74 to > 50 × 10⁶ daltons, three of which were giant extrachromosomal DNA elements. B. thuringiensis var. sotto contained one giant extrachromosomal DNA element with a molecular size of about 23.5 × 10⁶ daltons and two lesser elements of 0.80 and 0.62 × 10⁶ daltons. B. thuringiensis var. finitimus harbored two giant DNA elements corresponding to >50 × 10⁶ daltons and two lesser bands with relative small size (0.98 and 0.97 × 10⁶ daltons). B. popilliae contained no giant extrachromosomal DNA elements but did contain two smaller elements corresponding to 4.45 and 0.58 × 10⁶ daltons. The possible use of extrachromosomal DNA elements that prove to be autonomous replicons for recombinant DNA studies is discussed.     

Effect of entomopathogens on Africanized Apis mellifera L. (Hymenoptera: Apidae)

This study aimed to evaluate the effect of commercially used entomopathogens on Africanized Apis mellifera L. (Hymenoptera: Apidae). Four bioassays were performed: 1) pulverized entomopathogens on A. mellifera; 2) entomopathogens sprayed on a smooth surface; 3) entomopathogens sprayed on soy leaves; and 4) entomopathogens mixed with candy paste (sugar syrup). Five treatments were prepared: sterile distilled water (control), distilled water sterilized with Tween^® 80 (0.01%), and the commercial entomopathogens Metarhizium anisopliae E9 (1.0 × 10⁹ conidia mL^?1), Beauveria bassiana PL63 (1.0 × 10⁸ conidia mL^?1) and Bacillus thuringiensis var. kurstaki HD-1 (3.0 × 10⁸ spores mL^?1). Each treatment consisted of five repetitions, with 20 workers per repetition, which were stored in a plastic box and, later, in a biological oxygen demand (B.O.D.) incubator (27 ± 2 °C, RH of 60% ± 10%, 12-h photophase). The mortality of the workers was evaluated from 1 h to 240 h, and the data were analyzed using Bayesian inference. The workers killed by the ingestion of candy paste contaminated with the pathogens (products) were randomly separated and selected for the removal of the midgut. Each midgut was fixed in Bouin's solution and prepared for histology. B. bassiana was verified to reduce the survival of A. mellifera workers in all bioassays. Moreover, M. anisopliae reduced the survival of A. mellifera workers directly sprayed, on a smooth surface and mixed with candy. B. thuringiensis reduced A. mellifera survival on a smooth surface and mixed with candy paste. However, its effects were lower than that observed by B. bassiana. The treatments with the biological products did not induce morphometric alterations in the midgut of A. mellifera.     

Biological characterization of two <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> strains toxic against <Emphasis Type="Italic">Spodoptera frugiperda</Emphasis>

Two Bacillus thuringiensis strains isolated from diseased Spodoptera frugiperda larvae collected in the northwest of Argentina were molecularly and phenotypically characterized. Insecticidal activity against Spodoptera frugiperda larvae was also determined. Both strains were highly toxic against first instar larvae. One strain (Bacillus thuringiensis LSM) was found to be even more toxic than the reference strain Bacillus thuringiensis var. kurstaki 4D1. This strong biological effect was represented by both a higher mortality which reached 90%, and a shorter LT₅₀. Molecular characterization showed that Bacillus thuringiensis LSM carried a cry gene profile identical to that of Bacillus thuringiensis var. kurstaki 4D1. Evaluation of length polymorphism of the intergenic transcribed spacers between the 16S and 23S rDNA genes revealed an identical pattern between native strains and Bacillus thuringiensis var. kurstaki 4D1. In contrast, phenotypic characterization allowed differentiation among the isolates by means of their extracellular esterase profiles. Lytic activity that would contribute to Bacillus thuringiensis effectiveness was also studied in both strains. Analyses like those presented in the current study are essential to identify the most toxic strains and to allow the exploitation of local biodiversity for its application in biological control programmes.     

Plant-mediated vulnerability of an insect herbivore to Bacillus thuringiensis in a plant-herbivore-pathogen system

Laboratory studies were performed to explore the effects of host-plant quality on the vulnerability of Plutella xylostella to Bacillus thuringiensis. P. xylostella were kept on different host plants, including Brassica pekinensis (Chinese cabbage) cv. Hero, Brassica oleracea var. botrytis (cauliflower) cv. Royal, and B. oleracea var. capitata (common cabbage) cv. Globe Master (white cabbage) and cv. Red Dynasty (red cabbage) for at least two generations. These host plants are considered as the high (Chinese cabbage), intermediate (cauliflower and white cabbage) and low-quality (red cabbage) hosts for P. xylostella. The vulnerability of the pest larvae was then tested using two formulation of B. thuringiensis var. kurstaki, including Biolarv^® and Biolep^®. The results demonstrated that the susceptibility of P. xylostella to B. thuringiensis was influenced by host-plant quality. Indeed, B. thuringiensis acted better on the pest fed on the low-quality host plant compared with that on the high-quality host plant. The interaction between the pathogen and plant quality/resistance resulted in more mortality of the pest larvae, implying a synergistic effect. From a pest management viewpoint, these findings may be promising for the integration of the pathogen and the low-quality/partially resistant host plants against P. xylostella in field studies.     

Selectivity evaluation of insecticides used to control tomato pests to Trichogramma pretiosum

The effects of the insecticides abamectin, acetamiprid, cartap and chlorpyrifos on larvae, pupae (within the host egg) and adults of the egg parasitoid Trichogramma pretiosum Riley (Hym.: Trichogrammatidae) were evaluated under laboratory conditions, using three standard tests described by IOBC. When sprayed on the immature stages of this parasitoid, cartap and chlorpyrifos proved to be the most harmful insecticides, affecting both the emergence success and parasitism capacity of this parasitoid, whereas abamectin and acetamiprid were selective. Abamectin was harmful to adults (residue test on glass plates), slightly harmful to larvae, and moderately harmful to pupae (sprayed on the immature stages within host eggs Anagasta kuehniella (Zeller)); acetamiprid was moderately harmful to adults, harmless to larvae, and slightly harmful to pupae; cartap was harmful to adults, moderately harmful to larvae and harmful to pupae; chlorpyrifos to adults, harmless to larvae and harmful to pupae.     

Correlation between alkaline activation of Bacillus thuringiensis var. kurstaki spores and crystal production

The spores of crystal-forming (Cry⁺) and non-crystal-forming (Cry^-) strains of Bacillus thuringiensis var. kurstaki and Bacillus cereus were tested for the ability to be activated by 0.1 m K₂CO₃ (pH 10). Only the spores of crystal-forming strains could be activated, and this phenotype was independent of whether crystals were present with the spores in the activation solution. The spores of a B. thuringiensis var. kurstaki strain that is temperature sensitive for protoxin accumulation could be activated by the alkaline solution when produced at the permissive temperature, whereas spores produced at the nonpermissive temperature were not activated. The results indicate that protoxin in the spore coat is responsible for the alkaline-activation phenotype and may serve an ecological function for the organism.     

Interactions of Bacillus thuringiensis bioinsecticides and the predatory stink bug Podisus nigrispinus to control Plutella xylostella

The diamondback moth (DBM), Plutella xylostella (L.), is a major pest of brassica crops worldwide. Control of this pest is difficult because it rapidly develops resistance to synthetic and biological insecticides and because of the effects of insecticides on its natural enemies. Podisus nigrispinus (Dallas) is a predator that feeds on its prey, as well as on the host plants of its prey, and is an important biological control agent of DBMs. The aim of this study was to determine the susceptibility of P. xylostella larvae to two bioinsecticides: the HD1 strain of Bacillus thuringiensis (B. thuringiensis var. kurstaki) and the commercial product Agree^® (B. thuringiensis var. aizawai CG 91). In addition, the impact of these bioinsecticides on the P. nigrispinus consumption of DBM larvae and phytophagy was evaluated. Both the HD1 strain and Agree^® caused 100% mortality in P. xylostella larvae. P. nigrispinus nymphs fed only with kale leaves (Brassica oleracea var. acephala) sprayed with water, the HD1 strain, or Agree^® did not complete their nymphal development. When prey was also available, P. nigiripinus fed on kale leaves to obtain water. Both nymphs and adults of P. nigrispinus consumed greater numbers of DBM larvae, and fed less on kale leaves, when sprayed with the HD1 strain or Agree^®. These results suggest a positive interaction of B. thuringiensis‐based products and the predator P. nigrispinus in the control of P. xylostella larvae.     

Immunological properties of entomocidal protein of Bacillus thuringiensis and its insecticidal activity

The immunological properties of the proteinaceous component of the parasporal crystal (δ-endotoxin) of Bacillus thuringiensis var. kurstaki were analyzed by rocket immunoelectrophoresis. Two antisera, one against the k-l-type crystal containing two components, and the other against the k-73-type crystal containing one component, were made in rabbits. The antigens consisting of purified and dissociated crystals were run in electrophoresis with these two antisera. The ratio between the two peak heights of precipitin lines, which were formed by the dissociated crystal of one B. thuringiensis isolate in two antisera, was compared with the ratios of other isolates under identical conditions. The difference in the ratio reflected a difference in the structure of the crystal component and correlated closely with the insecticidal activity spectrum. This method can be used to evaluate a newly isolated B. thuringiensis, and it can further differentiate the isolates which have been classified as one serotype.     

Effects of Bacillus thuringiensis var. kurstaki δ-endotoxin on isolated lepidopteran mitochondria

An investigation was undertaken to determine the effects of Bacillus thuringiensis var. kurstaki δ-endotoxin on mitochondria isolated from Bombyx mori midgut epithelium. Using manometric and colorimetric techniques, the investigation revealed that toxic polypeptides had stimulatory effects on mitochondria oxygen uptake and inhibitory effects on ATP production. These results indicated that B. thuringiensis δ-endotoxin could act as an uncoupler of oxidative phosphorylation. Loss of ATP production caused by the action of the δ-endotoxin would lead to metabolic imbalance and possible cell death.     

Toxic effects of spore/crystal ratios of Bacillus thuringiensis on European corn borer larvae

Bioassays to determine LC₅₀ values of spores and crystals of four varieties of Bacillus thuringiensis grown on nutrient agar plates were carried out against neonate and 6-day-old European corn borer, Ostrinia nubilalis, larvae. The four bacterial varieties were equally toxic against the neonates, but only B. thuringiensis var. kenyae, var. galleriae, and var. kurstaki were toxic to 6-day-old larvae. B. thuringiensis var. tolworthi was inactive against 6-day-old larvae. Different ratios of pure spores and crystals of the bacteria also were tested against neonate and 6-day-old larvae. Pure spores are not pathogenic to neonates or 6-day-old larvae. Pure crystals were toxic to both ages of the larvae, but a combination of spores and crystals was necessary for maximum larval mortality.     

Effect of insecticides on the mortalities of three whitefly parasitoid species, Eretmocerusmundus, Eretmoceruseremicus and Encarsiaformosa (Hymenoptera: Aphelinidae)

The toxicities of 24 insecticides for the biological control of whiteflies were evaluated for Eretmocerus mundus (Mercet), Eretmocerus eremicus Rose and Zolnerowich and Encarsia formosa Gahan using the residual film method (for adults) and the dipping method (for pupae). Mortalities from insect growth regulators (IGRs) (flufenoxuron and lufenuron), Bacillus thuringiensis (Bt), pymetrozine and sulfur were <30% for both pupae and adults of all three species, indicating that the parasitoids were not seriously affected by these insecticides. Neonicotinoids (acetamiprid, clothianidin, dinotefuran, imidacloprid and nitenpyram), synthetic pyrethroids (etofenprox and permethrin), organophosphates (acephate and fenitrothion), chlorphenapyr, emamectin benzoate, spinosad and tolfenpyrad were seriously harmful (100% mortality) and acaricides (chinomethionat, milbemectin and pyridaben) were moderately harmful or seriously harmful to adult parasitoids (leading to mortalities of >92%). For each insecticide, the mortality of pupae was generally lower than that of adults, even though the toxicity classification for the two groups was similar. The results indicate that IGRs, Bt, pymetrozine and sulfur are relatively harmless, and are compatible with the use of parasitoids to help control whiteflies for integrated pest management in greenhouses.     

Resistance to Toxins from Bacillus thuringiensis subsp. kurstaki Causes Minimal Cross-Resistance to B. thuringiensis subsp. aizawai in the Diamondback Moth (Lepidoptera: Plutellidae)

Repeated exposure in the field followed by laboratory selection produced 1,800- to >6,800-fold resistance to formulations of Bacillus thuringiensis subsp. kurstaki in larvae of the diamondback moth, Plutella xylostella. Four toxins from B. thuringiensis subsp. kurstaki [CryIA(a), CryIA(b), CryIA(c), and CryIIA] caused significantly less mortality in resistant larvae than in susceptible larvae. Resistance to B. thuringiensis subsp. kurstaki formulations and toxins did not affect the response to CryIC toxin from B. thuringiensis subsp. aizawai. Larvae resistant to B. thuringiensis subsp. kurstaki showed threefold cross-resistance to formulations of B. thuringiensis subsp. aizawai containing CryIC and CryIA toxins. This minimal cross-resistance may be caused by resistance to CryIA toxins shared by B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. aizawai.     

Comparative selectivity of pesticides used in greenhouses,on the aphid parasitoid Aphidius colemani (Hymenoptera: Braconidae)

A series of bioassays were conducted under laboratory conditions to determine the relative toxicities of various pesticides (acetamiprid, cypermethrin, chlorantraniliprole and emamectin benzoate, Bacillus thuringiensis var. kurstaki and Helicoverpa armigera nucleopolyhedrovirus, copper oxychloride, iprodione, mandipropamid, a mixture of propamocarb?+?fluopicolide and mixture of fludioxonil?+?cyprodinil) on Aphidius colemani adults and mummies, as well as sublethal effects on female fecundity. Cypermethrin was highly toxic to pupa of A. colemani within host mummies. Acetamiprid, cypermethrin, emamectin benzoate, a mixture of propamocarb?+?fluopicolide and mixture of fludioxonil?+?cyprodinil were also highly toxic to A. colemani adults (92–100% mortality at 48?h post treatment). Mandipropamid, iprodione and copper oxycloride treatments significantly reduced fecundity of the female parasitoids. In contrast B. thuringiensis var. kurstaki, H. armigera nucleopolyhedrovirus and chlorantraniliprole were harmless (<30% mortality) to the parasitoid species tested according to International Organisation for Biological Control toxicity classification and are likely to be compatible with integrated pest management programmes.