ACE inhibitory activity in enzymatic hydrolysates of insect protein

In this paper, ACE inhibitory activity in insect protein hydrolyzed by various enzymes (gastrointestinal proteases, alcalase, and thermolysin) is reported for the first time. Four insects of different insect orders were tested: Spodoptera littoralis (Lepidoptera), Bombyx mori (Lepidoptera), Schistocerca gregaria (Orthoptera), and Bombus terrestris (Hymenoptera). ACE inhibitory activity was measured by two different methods: a spectrophotometric method using FAPGG (2-furanacryloyl-phenylalanyl-glycyl-glycine) as substrate, and an HPLC method using dansyltriglycine (DTG) as substrate. Hydrolysis of the insect protein resulted in an increased ACE inhibitory activity. Overall, the highest ACE inhibitory activity was obtained after gastrointestinal digestion. These results suggest a role for insect protein as antihypertensive component in functional foods and nutraceuticals. Furthermore, the ACE inhibitory activity differed according to the method used. As a consequence, there is a need to standardize methodologies to evaluate ACE inhibitory activity.     

Miniature-dispenser-based bioassay to evaluate the compatibility of powder formulations used in an entomovectoring approach

BACKGROUND: Entomovectoring as a plant protection strategy demands the design of an appropriate bioassay to assess the risks of potential side effects of the powder formulations in the dispenser towards the vectoring insect. This study reports on the development of a laboratory miniature‐dispenser‐based bioassay. This bioassay system was used to investigate the compatibility of five model products, Prestop‐Mix, Signum, kaolin, wheat flour and cellulose, with the bumblebee, Bombus terrestris L. RESULTS: The laboratory one‐way miniature‐dispenser bioassay showed that the fungicides and the carrier/diluent kaolin caused a worker mortality of > 70% after 5 weeks of exposure, while worker loss with wheat flour and cellulose was no higher than in the blank control (i.e. empty miniature dispenser) (<25%). The laboratory two‐way miniature‐dispenser bioassay comprised separated passageways and demonstrated that only kaolin was toxic (89 ± 11%). These results were also confirmed in a flight‐cage experiment. In addition, a negative effect was observed against reproduction/colony development when nests were exposed to kaolin (P < 0.05) in the two‐way miniature‐dispenser and flight‐cage bioassays. CONCLUSIONS: In the context of entomovectoring technology, the developed laboratory two‐way miniature‐dispenser bioassay gives a reliable prediction of the hazards associated with powder products. Additionally, the present data indicate the possibility of using cellulose and kaolin as respective negative and positive control carriers/diluents in future risk assessment experiments. Overall, the results show that, apart from kaolin, the tested fungicides and carriers/diluents are safe to be used with B. terrestris. Copyright © 2011 Society of Chemical Industry     

The non‐target impact of spinosyns on beneficial arthropods

Spinosyn‐based products, mostly spinosad, have been widely recommended by extension specialists and agribusiness companies; consequently, they have been used to control various pests in many different cropping systems. Following the worldwide adoption of spinosad‐based products for integrated and organic farming, an increasing number of ecotoxicological studies have been published in the past 10 years. These studies are primarily related to the risk assessment of spinosad towards beneficial arthropods. This review takes into account recent data with the aim of (i) highlighting potentially adverse effects of spinosyns on beneficial arthropods (and hence on ecosystem services that they provide in agroecosystems), (ii) clarifying the range of methods used to address spinosyn side effects on biocontrol agents and pollinators in order to provide new insights for the development of more accurate bioassays, (iii) identifying pitfalls when analysing laboratory results to assess field risks and (iv) gaining increasing knowledge on side effects when using spinosad for integrated pest management (IPM) programmes and organic farming. For the first time, a thorough review of possible risks of spinosad and novel spinosyns (such as spinetoram) to beneficial arthropods (notably natural enemies and pollinators) is provided. The acute lethal effect and multiple sublethal effects have been identified in almost all arthropod groups studied. This review will help to optimise the future use of spinosad and new spinosyns in IPM programmes and for organic farming, notably by preventing the possible side effects of spinosyns on beneficial arthropods. Copyright © 2012 Society of Chemical Industry     

Selectivity of diacylhydrazine insecticides to the predatory bug Orius laevigatus: in vivo and modelling/docking experiments

BACKGROUND: Knowledge of pesticide selectivity to natural enemies is necessary for a successful implementation of biological and chemical control methods in integrated pest management (IPM) programmes. Diacylhydrazine (DAH)‐based ecdysone agonists, also known as moulting‐accelerating compounds (MACs), are considered to be a selective group of insecticides, and their compatibility with predatory Heteroptera, which are used as biological control agents, is known. However, their molecular mode of action has not been explored in beneficial insects such as Orius laevigatus (Fieber) (Hemiptera: Anthocoridae). RESULTS: In this project, in vivo toxicity assays demonstrated that the DAH‐based RH‐5849, tebufenozide and methoxyfenozide have no toxic effect against O. laevigatus. The ligand‐binding domain (LBD) of the ecdysone receptor (EcR) of O. laevigatus was sequenced, and a homology protein model was constructed that confirmed a cavity structure with 12 α‐helices, harbouring the natural insect moulting hormone 20‐hydroxyecdysone. However, docking studies showed that a steric clash occurred for the DAH‐based insecticides owing to a restricted extent of the ligand‐binding cavity of the EcR of O. laevigatus. CONCLUSIONS: The insect toxicity assays demonstrated that MACs are selective for O. laevigatus. The modelling/docking experiments are indications that these pesticides do not bind with the LBD‐EcR of O. laevigatus and support the supposition that they show no biological effects in the predatory bug. These data help in explaining the compatible use of MACs together with predatory bugs in IPM programmes. Copyright © 2012 Society of Chemical Industry     

Compatibility of traditional and novel acaricides with bumblebees (Bombus terrestris): a first laboratory assessment of toxicity and sublethal effects

BACKGROUND: This project assessed the potential hazards of different classical and novel acaricides against an important non‐target and beneficial insect for the pollination of wild flowers and cultivated crops, the bumblebee Bombus terrestris (L). Twenty‐three acaricides used commercially in the control of phytophagous mites (Acari) were tested in greenhouses and/or the open field. Side effects included acute mortality and also sublethal effects on nest reproduction. The different compounds were administered in the laboratory via three different worst‐case field scenario routes of exposure: dermal contact and orally via the drinking of treated sugar water and via treated pollen. The compounds were tested at their respective maximum field recommended concentration (MFRC), and, when strong lethal effects were observed, a dose–response assay with a dilution series of the MFRC was undertaken to calculate LC₅₀ values. RESULTS: From the different acaricide classes, several chemistries caused high levels of acute toxicity in bumblebee workers, especially bifenthrin and abamectin which resulted in 100% mortality by contact. In addition, several acaricides tested were found to have a detrimental effect on drone production. For oral exposures via treated sugar water, the dose–response assay showed the LC₅₀ values for abamectin, bifenazate, bifenthrin and etoxazole to be 1/15 MFRC (1.17 mg AI L^?1), 1/10 MFRC (9.6 mg AI L^?1), 1/83 MFRC (0.36 mg AI L^?1) and 1/13 MFRC (4.4 mg AI L^?1) respectively, indicating that their use should be carefully evaluated. CONCLUSION: Overall, the results suggest that most of the acaricides tested are compatible with bumblebees, with the exceptions of abamectin, bifenazate, bifenthrin and etoxazole. However, the risks also depended on the type of treatment. As a result, the sugar water treatment seems to present the worst‐case situation of exposure, indicating that this approach is suitable for determining the hazards of pesticides against bumblebees. Finally, it is suggested that future tier testing under more field‐related conditions is required for a final decision of their risks. Copyright © 2010 Society of Chemical Industry     

Monitoring of beet armyworm resistance to spinosad and methoxyfenozide in Mexico

BACKGROUND: Resistance to spinosad and methoxyfenozide has been studied in several insect pests, but there is a lack of information on Spodoptera exigua (Hübner) in Mexico. Therefore, evidence for the development of resistance in this pest to both compounds was examined. The effects of methoxyfenozide on reproductive parameters of S. exigua adults were also determined.RESULTS: Third instars from a field population were exposed for 24 h to the LC(50) of spinosad or methoxyfenozide for over six generations (G(2)-G(7)). No significant reduction in susceptibility to either compound was detected for up to five generations. In G(7), LC(50) values for insects exposed to spinosad and methoxyfenozide were respectively 2.75-fold and 1.25-fold greater than for G(1) larvae. Oral treatment with methoxyfenozide reduced the fecundity and fertility of G(7) adults, confirming sublethal effects on reproduction. Finally, five populations (Se-La Floriza, Se-Lazareto, Se-Bachigualato, Se-Los Agustinos and Se-Villa de Arista) of S. exigua were collected from fields in three states of Mexico for resistance monitoring to spinosad and methoxyfenozide. With the exception of Se-Villa de Arista, the other populations showed significant resistance to spinosad, with resistance ratios between 16- and 37-fold, compared with a susceptible laboratory colony. In contrast, only one population (Se-Lazareto) showed significant resistance to methoxyfenozide (13-fold).CONCLUSION: Resistance management programmes should be established, particularly in areas where S. exigua has developed resistance to spinosad. Copyright (c) 2008 Society of Chemical Industry.     

Bombus terrestris as pollinator‐and‐vector to suppress Botrytis cinerea in greenhouse strawberry

BACKGROUND: Bombus terrestris L. bumblebees are widely used as commercial pollinators, but they might also be of help in the battle against economically important crop diseases. This alternative control strategy is referred to as pollinator‐and‐vector technology. The present study was designed to investigate the capacity of B. terrestris to fulfil this role in greenhouse strawberry flowers, which were manually inoculated with a major plant pathogen, the grey mould Botrytis cinerea Pers.: Fr. A model microbiological control agent (MCA) product Prestop‐Mix was loaded in a newly developed two‐way bumblebee dispenser, and, in addition, the use of the diluent Maizena‐Plus (corn starch) was tested. RESULTS: Importantly, loading of the MCA caused no adverse effects on bumblebee workers, with no loss of survival or impairment of flight activity of the workers during the 4 week flowering period. Secondly, vectoring of Prestop‐Mix by bumblebees resulted in a higher crop production, as 71% of the flowers developed into healthy red strawberries at picking (preharvest yield) as compared with 54% in the controls. In addition, these strawberries were better protected, as 79% of the picked berries remained free of B. cinerea after a 2 day incubation (post‐harvest yield), while this percentage was only 43% in the control. Overall, the total yield (preharvest × post‐harvest) was 2–2.5 times higher than the total yield in the controls (24%) in plants exposed to bumblebees vectoring Prestop‐Mix. Thirdly, the addition of the diluent Maizena‐Plus to Prestop‐Mix at 1:1 (w/w) resulted in a similar yield to that of Prestop‐Mix used alone, and in no negative effects on the bumblebees, flowers and berries. CONCLUSIONS: This greenhouse study provides strong evidence that B. terrestris bumblebees can vector a MCA to reduce B. cinerea incidence in greenhouse strawberries, resulting in higher yields. Similar yields obtained in the treatments with Prestop‐Mix and Prestop‐Mix + Maizena‐Plus suggest an equally efficient dissemination of the biocontrol agent into the flowers with only half the initial concentration of Prestop‐Mix, which illustrates the importance of the diluent. Copyright © 2011 Society of Chemical Industry     

A laboratory evaluation to determine the compatibility of microbiological control agents with the pollinator Bombus terrestris

BACKGROUND: This study was undertaken to identify any potential adverse side effects of the use of seven microbiological control agents (MCAs) on the bumblebee, Bombus terrestris L., in the context of combined use in integrated pest management (IPM). AQ10^® (Ampelomyces quisqualis), Binab‐T‐vector^® (Hypocrea parapilulifera + T. atroviride; 1/1), Prestop‐Mix^® (Gliocladium catenulatum J1446), Serenade^® (Bacillus subtilis QST713), Trianum‐P^® (Trichoderma harzianum T22), Botanigard^® (Beauveria bassiana GHA) and Granupom^® (Cydia pomonella granulovirus), comprising five biofungicides and two bioinsecticides, were investigated. Bumblebee workers were exposed under laboratory conditions to each MCA at its maximum field recommended concentration (MFRC) via three different routes of exposure: dermal contact and orally via either treated sugar water or pollen. RESULTS: The tested MCAs were found to be safe for workers of B. terrestris, with the exception of Botanigard^® and Serenade^®. Exposure to Botanigard^® via contact at its MFRC caused 92% mortality after 11 weeks, while the 1/10 MFRC killed 46% of exposed workers. For Serenade^®, topical contact and oral delivery via sugar water resulted in 88 and 100% worker mortality respectively. With lower concentrations (1/2, 1/5 and 1/10 MFRC) the toxicity decreased, but the effect depended on the route of exposure. In addition to lethal effects, nests were also evaluated for sublethal effects after treatment with the seven MCAs at their respective MFRCs over 11 weeks. In these bioassays, only Botanigard^® and Serenade^® gave rise to a significant (P < 0.05) decrease in drone production. Sublethal effects on foraging behaviour were also evaluated, and only Botanigard at its MFRC delivered via treated sugar water induced negative effects. CONCLUSION: The results demonstrated that most of the MCAs tested can be considered safe for use in combination with B. terrestris, based on the International Organisation for Biological Control of Noxious Animals and Plants (IOBC) classification. However, some can be harmful, such as the biofungicide Serenade^® and the bioinsecticide Botanigard^®. Therefore, it is recommended that all should be tested before use in combination with pollinators. In this context, it is also advisable that these MCAs should be evaluated in more realistic field situations for the assessment of potentially deleterious effects on foraging behaviour. Copyright © 2009 Society of Chemical Industry     

The effect of mass-flowering orchards and semi-natural habitat on bumblebee colony performance

Landscape Ecology - Bumblebees are important pollinators for agricultural crops and wild plants. However, agricultural intensification and loss of semi-natural habitat may have adverse effects on...