Insect Food Products in the Western World: Assessing the Potential of a New 'Green' Market.

Although two billion people already eat insects in the world and the benefits of edible insects are well known, these 'green' sources of protein are neither treated as conventional food products nor widely incorporated into Western diets. Using a school-based investigation surveying 161 children, aged 6-15, and 114 of their parents in London, and an online consumer survey with mainly British and French consumers (N = 1,020), this research provides insights into the potential of the insect market in the West. This work supports the idea that incorporating insect food into our diets makes not only environmental but also business sense. A nonnegligible segment of the population surveyed is willing to pay for mealworm minced meat and young children and pre-teens could represent a substantial market segment, as yet unexplored. This analysis points to multiple marketing strategies, such as early exposure, education, reducing the visibility of insect parts, celebrity endorsement, or peer-to-peer marketing, all of which could facilitate the adoption of insect food in the 'mainstream' arena, according to the consumer segment being targeted. Generalizations from these results are restricted to an educated and youthful subset of the potential consumer pool and further work remains to understand the patterns of Western consumer acceptance for the range of insect foods.

Stimulating Anopheles gambiae swarms in the laboratory: application for behavioural and fitness studies.

BACKGROUND: Male Anopheles mosquitoes that swarm rely in part on features of the environment including visual stimuli to locate swarms. Swarming is believed to be the primary behaviour during which mating occurs in the field, but is not a common behaviour in the laboratory. Features that stimulate male Anopheles gambiae G3 strain swarming were created in novel large indoor cages. METHODS: The following visual features were tested in all combinations to determine which were important for swarm formation. Large cages and fading ceiling lights at dusk alone did not stimulate swarming while a dark foreground and contrasting illuminated background with a contrasting landmark stimulated and localized swarm formation during artificial twilight. Given the need to test transgenic strains in as natural a setting as possible, in this study it was investigated whether induced swarm behaviour and cage size would affect relative mating performance of wild-type and transgenic ß2Ppo1 and ß2Ppo2 A. gambiae sexually sterile males. RESULTS: Even using a mosquito colony that has been in laboratory culture for 39 years, swarming behaviour was induced by this novel arrangement. The presence of swarming stimuli was associated with an increase in insemination frequency from 74.3 to 97.7% in large cages. Transgenic males showed a lower competitiveness in large cages compared to small cages regardless of the presence of swarming stimuli. CONCLUSIONS: The results of the present study are discussed in view of the progressive evaluation of genetically modified A. gambiae strains and the potential applications of reproducing swarms in controlled conditions to dissect the mating behaviour of this species and the mechanisms controlling it.

Could species-focused suppression of Aedes aegypti, the yellow fever mosquito, and Aedes albopictus, the tiger mosquito, affect interacting predators? An evidence synthesis from the literature.

The risks of Aedes aegypti and Aedes albopictus nuisance and vector-borne diseases are rising and the adverse effects of broad-spectrum insecticide application have promoted species-specific techniques, such as sterile insect technique (SIT) and other genetic strategies, as contenders in their control operations. When specific vector suppression is proposed, potential effects on predators and wider ecosystem are some of the first stakeholder questions. These are not the only Aedes vectors of human diseases, but are those for which SIT and genetic strategies are of most interest. They vary ecologically and in habitat origin, but both have behaviorally human-adapted forms with expanding ranges. The aquatic life stages are where predation is strongest due to greater resource predictability and limited escape opportunity. These vectors' anthropic forms usually use ephemeral water bodies and man-made containers as larval habitats; predators that occur in these are mobile, opportunistic and generalist. No literature indicates that any predator depends on larvae of either species. As adults, foraging theory predicts these mosquitoes are of low profitability to predators. Energy expended hunting and consuming will mostly outweigh their energetic benefit. Moreover, as adult biomass is mobile and largely disaggregated, any predator is likely to be a generalist and opportunist. This work, which summarizes much of the literature currently available on the predators of Ae. aegypti and Ae. albopictus, indicates it is highly unlikely that any predator species depends on them. Species-specific vector control to reduce nuisance and disease is thus likely to be of negligible or limited impact on nontarget predators. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A systematic review of the leaf traits considered to contribute to removal of airborne particulate matter pollution in urban areas.

Global urban planning has promoted green infrastructure (GI) such as street trees, shrubs or other greenspace in order to mitigate air pollution. Although considerable attention has been paid to understanding particulate matter (PM) deposition on GI, there has been little focus on identifying which leaf traits might maximise airborne PM removal. This paper examines existing literature to synthesize the state of knowledge on leaf traits most relevant to PM removal. We systematically reviewed measurement studies that evaluated particulate matter accumulated on leaves on street trees, shrubs green roofs, and green walls, for a variety of leaf traits. Our final selection included 62 papers, most from field studies and a handful from wind tunnel studies. The following were variously promoted as useful traits: coniferous needle leaves; small, rough and textured broadleaves; lanceolate and ovate shapes; waxy coatings, and high-density trichomes. Consideration of these leaf traits, many of which are also associated with drought tolerance, may help to maximise PM capture. Although effective leaf traits were identified, there is no strong or consistent evidence to identify which is the most influential leaf trait in capturing PM. The diversity in sampling methods, wide comparison groups and lack of background PM concentration measures in many studies limited our ability to synthesize results. We found that several ancillary factors contribute to variations in the accumulation of PM on leaves, thus cannot recommend that selection of urban planting species be based primarily on leaf traits. Further research into the vegetation structural features and standardization of the method to measure PM on leaves is needed.

Eave and swarm collections prove effective for biased captures of male Anopheles gambiae mosquitoes in Uganda.

BACKGROUND: Traditional malaria vector sampling techniques bias collections towards female mosquitoes. Comprehensive understanding of vector dynamics requires balanced vector sampling of both males and females. Male mosquito sampling is also necessary for population size estimations by male-based mark-release-recapture (MRR) studies and for developing innovations in mosquito control, such as the male-targeted sterile insect technique and other genetic modification approaches. This study evaluated a range of collection methods which show promise in providing a more equal, or even male-biased, sex representation in the sample. RESULTS: Swarms were found at all study sites and were more abundant and larger at the peak of the wet season. Swarm sampling caught the most males, but when man/hour effort was factored in, sampling of eaves by aspiration was the more efficient method and also provided a representative sample of females. Grass-roofed houses were the most productive for eave collections. Overall few mosquitoes were caught with artificial resting traps (clay pots and buckets), although these sampling methods performed better at the start of the wet season than at its peak, possibly because of changes in mosquito ecology and an increased availability of natural resting sites later in the season. Aspiration of bushes was more productive at the peak of the wet season than at the start. CONCLUSIONS: The results of this study demonstrate that eave aspiration was an efficient and useful male mosquito collection method at the study sites and a potentially powerful aid for swarm location and MRR studies. The methods evaluated may together deliver more sex-balanced mosquito captures and can be used in various combinations depending on the aims and ecological parameters of a given study.

Sterile Insect Technique (SIT) against Aedes Species Mosquitoes: A Roadmap and Good Practice Framework for Designing, Implementing and Evaluating Pilot Field Trials.

Aedes albopictus and Aedes aegypti are invasive mosquito species that impose a substantial risk to human health. To control the abundance and spread of these arboviral pathogen vectors, the sterile insect technique (SIT) is emerging as a powerful complement to most commonly-used approaches, in part, because this technique is ecologically benign, specific, and non-persistent in the environment if releases are stopped. Because SIT and other similar vector control strategies are becoming of increasing interest to many countries, we offer here a pragmatic and accessible 'roadmap' for the pre-pilot and pilot phases to guide any interested party. This will support stakeholders, non-specialist scientists, implementers, and decision-makers. Applying these concepts will ensure, given adequate resources, a sound basis for local field trialing and for developing experience with the technique in readiness for potential operational deployment. This synthesis is based on the available literature, in addition to the experience and current knowledge of the expert contributing authors in this field. We describe a typical path to successful pilot testing, with the four concurrent development streams of Laboratory, Field, Stakeholder Relations, and the Business and Compliance Case. We provide a graphic framework with criteria that must be met in order to proceed.

Auditing Preparedness for Vector Control Field Studies.

The value of baseline entomological data to any future area-wide release campaign relies on the application of consistent methods to produce results comparable across different times and places in a stepwise progression to larger releases. Traditionally, standard operating procedures (SOPs) and operational plans support this consistency and, thus, the validity of emergent data. When release plans include transgenic mosquitoes for vector control or other novel beneficial insects, additional factors come into play such as biosafety permits, stakeholder acceptance, and ethics approval, which require even greater coordination and thoroughness. An audit approach was developed to verify the correct use of SOPs and appropriate performance of tasks during mosquito mark, release, recapture (MRR) studies. Audit questions matched SOPs, permit terms and conditions, and other key criteria, and can be used to support subsequent "spot check" verification by field teams. An external team of auditors, however, was found to be effective for initial checks in this example before the use of a transgenic strain of laboratory mosquitoes. We recommend similar approaches for field studies using release of novel beneficial insects, to ensure useful and valid data as an outcome and to support confidence in the rigor of the step-wise process.

Measuring and reducing biofilm in mosquito rearing containers.

BACKGROUND: Mosquito rearing containers contain organic-rich water that nourishes numerous bacteria, some of which are capable of forming biofilms. Biofilm is broadly an extracellular polymeric matrix (EPS) in which living bacteria occur, and the accumulation of biofilm is possible during routine stock-keeping as most of these containers are re-used. Whether biofilm has an effect on the mosquito rearing is not a question that has been investigated, nor have measures to reduce biofilm in this context been systematically studied. METHODS: We measured biofilm accumulation in standard rearing containers by staining with crystal violet and determining the OD using a spectrophotometer. We also treated rearing containers with 0.1% sodium hypochlorite to determine its effectiveness in reducing biofilm abundance. Lastly, we performed an analysis of the relationship between the occurrence of biofilm and the likelihood of microbial blooms that were associated with larval death during trials of larval diets. RESULTS: We observed that soaking rearing containers overnight in 0.1% sodium hypochlorite greatly reduced biofilm, but we observed no relationship between the use of containers that had not been treated with bleach and subsequent microbial blooms. CONCLUSIONS: Larva rearing leaves detectable biofilm. While we were unable to correlate microbial blooms with the presence of biofilm, as a precaution, we recommend that plastic containers that are re-used be treated with 0.1% sodium hypochlorite occasionally.

Surviving the Journey: Comparisons of Temperature-Stabilizing Materials for Living Arthropod Shipments.

Shipments of living mosquitoes and other arthropods require temperatures that are within a range that is compatible with their health and survival. In addition to express shipping and insulated containers, shipments often include materials that either store heat (i.e., have thermal mass) or otherwise stabilize the temperature. In this paper, we present the results of comparisons of thermal mass and phase change materials to stabilize the temperature under various conditions. We compared a rigid foam refrigerant and a number of phase change materials to bubble wrap for their capacity to moderate temperature change by measuring the temperatures in standard uninsulated shipping containers during exposure to high (37°C), cold (4°C), and freezing (-20°C) temperatures. We make recommendations for shipments depending on the ambient conditions that are expected to be experienced en route.

Pragmatic selection of larval mosquito diets for insectary rearing of Anopheles gambiae and Aedes aegypti.

Larval mosquitoes are aquatic omnivorous scavengers which scrape food from submerged surfaces and collect suspended food particles with their mouth brushes. The composition of diets that have been used in insectaries varies widely though necessarily provides sufficient nutrition to allow colonies to be maintained. Issues such as cost, availability and experience influence which diet is selected. One component of larval diets, essential fatty acids, appears to be necessary for normal flight though deficiencies may not be evident in laboratory cages and are likely more important when mosquitoes are reared for release into the field in e.g. mark-release-recapture and genetic control activities. In this study, four diets were compared for rearing Anopheles gambiae and Aedes aegypti, all of which provide these essential fatty acids. Two diets were custom formulations specifically designed for mosquitoes (Damiens) and two were commercially available fish foods: Doctors Foster and Smith Koi Staple Diet and TetraMin Plus Flakes. Development rate, survival, dry weight and adult longevity of mosquitoes reared with these four diets were measured. The method of presentation of one diet, Koi pellets, was additionally fed in two forms, pellets or a slurry, to determine any effect of food presentation on survival and development rate. While various criteria might be selected to choose 'the best' food, the readily-available Koi pellets resulted in development rates and adult longevity equal to the other diets, high survival to the adult stage and, additionally, this is available at low cost.

Large-cage assessment of a transgenic sex-ratio distortion strain on populations of an African malaria vector.

BACKGROUND: Novel transgenic mosquito control methods require progressively more realistic evaluation. The goal of this study was to determine the effect of a transgene that causes a male-bias sex ratio on Anopheles gambiae target populations in large insectary cages. METHODS: Life history characteristics of Anopheles gambiae wild type and Ag(PMB)1 (aka gfp124L-2) transgenic mosquitoes, whose progeny are 95% male, were measured in order to parameterize predictive population models. Ag(PMB)1 males were then introduced at two ratios into large insectary cages containing target wild type populations with stable age distributions and densities. The predicted proportion of females and those observed in the large cages were compared. A related model was then used to predict effects of male releases on wild mosquitoes in a west African village. RESULTS: The frequency of transgenic mosquitoes in target populations reached an average of 0.44 ± 0.02 and 0.56 ± 0.02 after 6 weeks in the 1:1 and in the 3:1 release ratio treatments (transgenic male:wild male) respectively. Transgenic males caused sex-ratio distortion of 73% and 80% males in the 1:1 and 3:1 treatments, respectively. The number of eggs laid in the transgenic treatments declined as the experiment progressed, with a steeper decline in the 3:1 than in the 1:1 releases. The results of the experiment are partially consistent with predictions of the model; effect size and variability did not conform to the model in two out of three trials, effect size was over-estimated by the model and variability was greater than anticipated, possibly because of sampling effects in restocking. The model estimating the effects of hypothetical releases on the mosquito population of a West African village demonstrated that releases could significantly reduce the number of females in the wild population. The interval of releases is not expected to have a strong effect. CONCLUSIONS: The biological data produced to parameterize the model, the model itself, and the results of the experiments are components of a system to evaluate and predict the performance of transgenic mosquitoes. Together these suggest that the Ag(PMB)1 strain has the potential to be useful for reversible population suppression while this novel field develops.

Comparison of Model Predictions and Laboratory Observations of Transgene Frequencies in Continuously-Breeding Mosquito Populations.

The persistence of transgenes in the environment is a consideration in risk assessments of transgenic organisms. Combining mathematical models that predict the frequency of transgenes and experimental demonstrations can validate the model predictions, or can detect significant biological deviations that were neither apparent nor included as model parameters. In order to assess the correlation between predictions and observations, models were constructed to estimate the frequency of a transgene causing male sexual sterility in simulated populations of a malaria mosquito Anopheles gambiae that were seeded with transgenic females at various proportions. Concurrently, overlapping-generation laboratory populations similar to those being modeled were initialized with various starting transgene proportions, and the subsequent proportions of transgenic individuals in populations were determined weekly until the transgene disappeared. The specific transgene being tested contained a homing endonuclease gene expressed in testes, I-PpoI, that cleaves the ribosomal DNA and results in complete male sexual sterility with no effect on female fertility. The transgene was observed to disappear more rapidly than the model predicted in all cases. The period before ovipositions that contained no transgenic progeny ranged from as little as three weeks after cage initiation to as long as 11 weeks.

Cryptic virulence and avirulence alleles revealed by controlled sexual recombination in pea aphids.

Although aphids are worldwide crop pests, little is known about aphid effector genes underlying virulence and avirulence. Here we show that controlling the genetics of both aphid and host can reveal novel recombinant genotypes with previously undetected allelic variation in both virulence and avirulence functions. Clonal F1 progeny populations were derived from reciprocal crosses and self-matings between two parental genotypes of pea aphid (Acyrthosiphon pisum) differing in virulence on a Medicago truncatula host carrying the RAP1 and RAP2 resistance genes. These populations showed Mendelian segregation consistent with aphid performance being controlled largely by a dominant virulence allele derived from only one parent. Altered segregation ratios on near-isogenic host genotypes differing in the region carrying RAP1 were indicative of additional heritable functions likely related to avirulence genes originating from both parents. Unexpectedly, some virulent F1 progeny were recovered from selfing of an avirulent parent, suggesting a reservoir of cryptic alleles. Host chlorosis was associated with virulence, whereas necrotic hypersensitive-like response was not. No maternal inheritance was found for any of these characteristics, ruling out sex-linked, cytoplasmic, and endosymbiotic factors. Our results demonstrate the tractability of dissecting the genetic basis of pest-host resistance mechanisms and indicate that the annual sexual cycle in aphids may lead to frequent novel genotypes with both increased and decreased virulence. Availability of genomes for both pest and host can facilitate definition of cognate gene-for-gene relationships, potentially leading to selection of crop genotypes with multiple resistance traits.