Can there be a common, risk-based framework for decisions around live insect trade?

A network of scientists involved in shipment of live insects has met and generated a series of articles on issues related to live insect transport. The network is diverse, covering large-scale commercial interests, government operated areawide control programmes, biomedical research and many smaller applications, in research, education and private uses. Many insect species have a record of safe transport, pose minimal risks and are shipped frequently between countries. The routine shipments of the most frequently used insect model organism for biomedical research, Drosophila melanogaster, is an example. Successful large scale shipments from commercial biocontrol and pollinator suppliers also demonstrate precedents for low-risk shipment categories, delivered in large volumes to high quality standards. Decision makers need access to more information (publications or official papers) that details actual risks from the insects themselves or their possible contaminants, and should propose proportionate levels of management. There may be harm to source environments when insects are collected directly from the wild, and there may be harm to receiving environments. Several risk frameworks include insects and various international coordinating bodies, with experience of guidance on relevant risks, exist. All stakeholders would benefit from an integrated overview of guidance for insect shipping, with reference to types of risk and categories of magnitude, without trying for a single approach requiring universal agreement. Proposals for managing uncertainty and lack of data for smaller or infrequent shipments, for example, must not disrupt trade in large volumes of live insects, which are already supporting strategic objectives in several sectors.

À l'occasion d'une réunion d'experts, un réseau de chercheurs travaillant sur l'expédition d'insectes vivants a produit un ensemble d'articles traitant des questions liées au transport des insectes vivants. Ce réseau est diversifié et représente un large éventail d'intérêts privés et de programmes de lutte biologique à grande échelle menés par les pouvoirs publics, en plus du secteur de la recherche biomédicale et de nombreux acteurs intervenant dans des applications de plus petite envergure relevant de la recherche, de l'enseignement ou du secteur privé. Un grand nombre d'espèces d'insectes sont transportées en toute sécurité avec un niveau de risque minimal, y compris lors des fréquentes expéditions internationales. Un exemple de ces expéditions régulières concerne l'espèce la plus utilisée par la recherche biomédicale en tant qu'organisme modèle, à savoir Drosophila melanogaster. Les exemples réussis d'expéditions à grande échelle provenant de fournisseurs d'agents de lutte biologique et de pollinisateurs produits par le secteur privé offrent également un relevé documenté des diverses catégories d'expéditions à faible risque pour des livraisons d'insectes vivants en grandes quantités et répondant à des normes de qualité élevées. Les décideurs politiques devraient pouvoir accéder à plus d'informations (à travers des publications ou des articles officiels) décrivant en détail les risques réels associés aux insectes eux-mêmes ou à leurs contaminants éventuels, et proposer en connaissance de cause des niveaux de gestion proportionnels à ces risques. La récolte d'insectes prélevés directement de la nature peut être dommageable aussi bien pour les environnements source que pour ceux de destination. Plusieurs cadres fondés sur le risque intègrent désormais les insectes dans leurs directives. Par ailleurs, nombre d'organismes internationaux de coordination ont acquis une expérience dans l'élaboration de lignes directrices face à ces risques. Il serait bénéfique pour toutes les parties prenantes de disposer d'une vue d'ensemble intégrée des directives applicables aux expéditions d'insectes, qui recense les différents types de risque et leurs ordres de grandeur sans se prononcer sur une approche unique qui nécessiterait une adhésion universelle. Ni les propositions visant à gérer l'incertitude ni l'insuffisance des données disponibles sur les expéditions d'insectes en petits nombres ou occasionnelles ne doivent perturber les échanges commerciaux d'insectes vivants en grandes quantités, échanges qui participent aujourd'hui aux objectifs stratégiques de nombreux secteurs.

Una red de científicos relacionados de un modo u otro con el transporte de insectos vivos mantuvo un encuentro y generó una serie de artículos en torno a la cuestión. En la red, muy heterogénea, convergen desde intereses comerciales de gran calado hasta programas públicos de lucha biológica en grandes territorios, pasando por la investigación biomédica y por numerosas aplicaciones de menor dimensión en ámbitos como la investigación, la enseñanza u otros usos privados. Muchas especies de insectos cuentan con un buen historial de seguridad en el transporte, presentan un riesgo mínimo y son expedidas a menudo de un país a otro. Buen ejemplo de ello son los envíos sistemáticos de ejemplares de Drosophila melanogaster, que es el insecto utilizado con más frecuencia como organismo modelo en la investigación biomédica. Otro precedente de envíos voluminosos cuyo transporte y entrega se ajusta a los más exigentes criterios de calidad lo sientan las remesas de grandes cantidades de polinizadores y agentes de control biológico remitidos por proveedores comerciales. Los responsables de adoptar decisiones deben disponer de más y más detallada información (publicaciones o documentos oficiales) sobre los riesgos reales derivados de los propios insectos o sus posibles contaminantes para proponer a partir de ahí medidas de gestión proporcionadas. Los perjuicios ambientales pueden darse tanto en origen (cuando hay captura salvaje de insectos, o sea recolección directa en el medio) como en destino. Hay varios sistemas de determinación del riesgo que incluyen a los insectos y también existen diversos organismos de coordinación internacional que ya tienen experiencia en sentar pautas sobre los riesgos en la materia. Todos los interlocutores del sector se beneficiarían de una visión global e integrada del transporte de insectos, que remita a diferentes tipos de riesgo y categorías de magnitud, sin necesidad de buscar un sistema único que exija consenso universal. Las propuestas para gestionar la incertidumbre y la falta de información en el caso de envíos infrecuentes o de pequeñas dimensiones, por ejemplo, no deben interferir en el transporte de grandes cantidades de insectos vivos, que ya está sirviendo a objetivos estratégicos en varios sectores.

Managing stakeholder concerns associated with releases of imported stock in insect control programmes.

A commitment to reducing pesticide use and the development of novel technologies are driving a renewed interest in insect-mediated pest and vector control programmes. Such programmes, along with conservation and pollination applications, lead to an increased transport volume of live insect stock. At release sites, concerns surrounding imported insects can be reduced by using local genotypes that have been mass-produced elsewhere. Remaining plausible concerns are likely to be centred on human factors (vector behaviour or capacity) and ecological factors (interacting species) and should be anticipated in the design of communication materials. Well-designed, locally relevant communication and engagement material is an important part of programme success. Stakeholder engagement is thus critical to reducing risks of perceived and plausible concerns affecting programme outcomes in an increasingly electronically connected world. Experience at release sites can help inform the design of accessible information useful at all stages of the transportation pathway. For transnationally transported insects, providing such information to specific stakeholders (e.g. courier companies and border authorities) will reduce the likelihood of delays, which can, in turn, affect the quality and mortality of the transported insects.

Les engagements pris en matière de réduction de l'utilisation de pesticides et les nouvelles technologies disponibles ont donné un nouvel essor aux programmes de contrôle des parasites et des vecteurs axés sur les insectes. Ces programmes, auxquels s'ajoutent d'autres applications destinées à favoriser la conservation et la pollinisation, se traduisent par un accroissement du volume de stocks d'insectes vivants transportés dans le monde. Dans les sites de destination des insectes transportés, les inquiétudes que peuvent susciter ces importations peuvent être tempérées en faisant appel à des génotypes locaux, produits en masse ailleurs puis réintroduits. D'autres préoccupations peuvent subsister, portant sur des facteurs humains (liées aux effets du comportement des vecteurs ou à leur capacité vectorielle) et écologiques (liées aux espèces en interaction) ; il convient d'anticiper ces inquiétudes lors de la conception des supports de communication. Des matériels de communication et de mobilisation bien conçus et prenant en compte le contexte local sont une composante importante de la réussite d'un programme. La participation des parties prenantes se révèle donc cruciale pour réduire le risque que des inquiétudes perçues et plausibles viennent affecter les résultats d'un programme dans un monde de plus en plus interconnecté par voie électronique. L'expérience acquise dans les sites de lâchers d'insectes peut contribuer à documenter la conception d'une information accessible, qui sera utile à chaque étape de la procédure de transport. Lorsqu'il s'agit d'insectes transportés au-delà des frontières nationales, la diffusion de ce type d'information à des intervenants spécifiques (par exemple, les sociétés de transport express et les autorités frontalières) permettra de limiter les circonstances entraînant des retards d'acheminement, car ceux-ci peuvent à leur tour affecter la qualité et la viabilité des insectes transportés.

El empeño de reducir el uso de plaguicidas y la aparición de tecnologías novedosas están alimentando un renovado interés por los programas de control de plagas y vectores mediante insectos. Este tipo de programas, junto con las aplicaciones de los insectos con fines de polinización o de protección ambiental, se traducen en el transporte de volúmenes crecientes de poblaciones de insectos vivos. Para apaciguar los recelos que despiertan los insectos importados en los sitios de suelta cabe recurrir al empleo de genotipos locales criados a gran escala en otro lugar. A la hora de concebir el material de comunicación es conveniente prever de antemano y tener en cuenta las suspicacias plausibles que aún puedan subsistir, que seguramente tendrán que ver con factores humanos (por ejemplo, preocupación por los posibles efectos del comportamiento o la capacidad vectorial del insecto introducido) o ecológicos (interacción con otras especies). El uso de material de comunicación y participación bien concebido y adaptado al contexto local es un ingrediente importante para el éxito de todo programa. La participación de las partes interesadas es pues fundamental para reducir los riesgos (ya sean imaginados o plausibles) que puedan afectar los resultados del programa, en un mundo que está cada vez más interconectado electrónicamente. La experiencia adquirida en los sitios de suelta puede ayudar a concebir material informativo accesible que sea de utilidad en todas las etapas del proceso de transporte. En el caso de los insectos transportados de un país a otro, el hecho de facilitar esta información a determinadas partes (como las empresas de transporte o las autoridades aduaneras) reducirá la probabilidad de demoras, cosa que a su vez incidirá en la calidad y la viabilidad de los insectos transportados.

Focal distribution of diflubenzuron resistance mutations in Culex pipiens mosquitoes from Northern Italy.

Insecticide resistance is a major threat for vector control and prevention of mosquito borne diseases. In the Culex pipiens mosquitoes, resistance against diflubenzuron (DFB) was firstly detected in Ravenna (Emilia-Romagna region, Northern Italy), in 2015. The resistant phenotypes were associated with two mutations, I1043 M and I1043 L, at the amino acid 1043 of the chitin synthase gene. In this study, we monitored the presence, frequency and geographical distribution of the DFB resistant mutations in Cx. pipiens populations from Northern Italy, and in populations from Greece and France. In the Emilia-Romagna region, the resistant mutations were detected in 20 out of the 30 populations analysed, reaching allelic frequencies over 70%. The presence and distribution of the resistance mutations was highly focal, with a clear pattern of increasing resistant allelic frequencies moving from the Western towards the Eastern provinces of Emilia-Romagna. Contrary to Italy, DFB resistant alleles were not detected in the Cx. pipiens mosquitoes sampled from Greece and France. Following statistical, literature and bibliographical database analyses on the history of DFB insecticide use in the study areas, we suggest that the selection pressures from the intense agricultural DFB applications occurring throughout the' 80-'90 s against orchard pests, followed, from 2000s onwards by mosquito control DFB applications, may account for the high mutation frequencies observed in the Cx. pipiens populations of the Eastern provinces of Emilia-Romagna. The findings are of major concern for public health in Italy and Europe, as DFB remains a very important insecticide used for controlling arbovirus mosquito vectors, where alternative larvicides are extremely limited.

"Gold" Pressed Essential Oil: An Essay on the Volatile Fragment from Citrus Juice Industry By-Products Chemistry and Bioactivity.

Present essay explores the potentials of Citrus juice industry's by-products as alternative bioactive natural products resources. Four crude Cold Pressed Essential Oils (CPEOs), derived from orange, lemon, grapefruit, and mandarin, were studied. All CPEOs were subjected to water distillation, in order to obtain the volatile fragment, which was further fractionated with respect to distillation period in two parts, concluding to eight samples. These samples along with the four original CPEOs were assessed in relation to their phytochemical content and their repellent and larvicidal properties against Asian Tiger Mosquito. The volatiles recovery rates ranged from 74% to 88% of the CPEO. Limonene presented a significant increase in all samples ranging from 8% to 52% of the respective CPEO's content and peaked in mandarin's 2nd volatile fragment which comprised 97% of the essential oil. The refinement process presented clear impacts on both bioassays: a significant increase in larvicidal potency was observed, annotated best by the improvement by 1100% and 1300% of the grapefruit volatile fractions; repellence testing provided only one significant result, the decrease of landings by 50% as a response to mandarin's second volatile fraction. The applied methodology thus may be considered for the improvement of Citrus juice industry's by-products chemistry and bioactivity.

Seasonal variation and bioactivity of the essential oils of two Juniperus species against Aedes (Stegomyia) albopictus (Skuse, 1894).

The seasonal variation in respect to the yield and chemical composition of 24 essential oils (EOs) isolated from various parts (leaves and fruits) of two indigenous Greece Juniperus species (family Cupressaceae), namely Juniperus drupacea and Juniperus phoenica, were determined by GC and GC/MS analysis. The larvicidal properties of these EOs were evaluated against 3rd and early 4th instar larvae of Aedes (Stegomyia) albopictus (Skuse, 1894) at one screening dose (29 mg L(-1)). Moreover, the repellent activity against adult mosquitoes was also evaluated at one screening dose. The analytical data indicated that the EOs mainly consisted of monoterpenes, mostly cyclic and only occasionally aliphatic and to a lesser percent diterpenes. The EOs yield was sharply increased when the plant material was subjected to pre-treatment before steam distillation. Finally, the influence of plant material collection period on their yield and chemical content was also determined. Bioactivity assessments indicated that three EOs possess very potent larvicidal properties and 12 EOs display significant repellent activities since they were proved to be "DEET-like." Therefore, they represent an inexpensive source of natural mixtures of larvicidal and repellent mixture of natural compounds, with potentials for application for utilization in mosquito control schemes in order to prevent the expansion of viral infections.

Economic appraisal of the public control and prevention strategy against the 2010 West Nile Virus outbreak in Central Macedonia, Greece.

OBJECTIVES: The aim of the present paper is to evaluate the economic efficiency of the public control and prevention strategies to tackle the 2010 West Nile Virus (WNV) outbreak in the Region of Central Macedonia, Greece. Efficiency is examined on the basis of the public prevention costs incurred and their potential in justifying the costs arising from health and nuisance impacts in the succeeding years. STUDY DESIGN: Economic appraisal of public health management interventions. METHODS: Prevention and control cost categories including control programmes, contingency planning and blood safety testing, are analyzed based on market prices. A separate cost of illness approach is conducted for the estimation of medical costs and productivity losses from 2010 to 2013 and for the calculation of averted health impacts. The averted mosquito nuisance costs to households are estimated on the basis of a contingent valuation study. Based on these findings, a limited cost-benefit analysis is employed in order to evaluate the economic efficiency of these strategies in 2010-2013. RESULTS: Results indicate that cost of illness and prevention costs fell significantly in the years following the 2010 outbreak, also as a result of the epidemic coming under control. According to the contingent valuation survey, the annual average willingness to pay to eliminate the mosquito problem in the study area ranged between 22 and 27 € per household. Cost-benefit analysis indicates that the aggregate benefit of implementing the previous 3-year strategy creates a net socio-economic benefit in 2013. However the spread of the WNV epidemic and the overall socio-economic consequences, had the various costs not been employed, remain unpredictable and extremely difficult to calculate. CONCLUSIONS: The application of a post epidemic strategy appears to be of utmost importance for public health safety. An updated well designed survey is needed for a more precise definition of the optimum prevention policies and levels and for the establishment of the various cost/benefit parameters.

Evaluation of the repellent effects of Nepeta parnassica extract, essential oil, and its major nepetalactone metabolite against mosquitoes.

The dichloromethane-methanol extract, the essential oil obtained by hydrodistillation from Nepeta parnassica, and the isolated 4aα,7α,7aß-nepetalactone were evaluated for their repellent effect against the mosquitoes Aedes (Stegomyia) cretinus Edwards and Culex pipiens pipiens biotype molestus Forskål. The chemical analysis of N. parnassica essential oil, dominated by oxygenated monoterpenes (87 %), revealed 4aα,7α,7aß-nepetalactone (36.8%), 1,8-cineole (25.5%), and 4aα,7ß,7aß-nepetalactone (11.1%) as the major constituents. The results of the insect bioassays showed that the essential oil and the dichloromethane-methanol extract of N. parnassica were very active against Aedes cretinus for up to 3 h and against Culex pipiens for up to 2 h post application. The isolated 4aα,7α,7aß-nepetalactone showed very high mosquito repellency for periods of at least 2 h against both species.

Evaluation of naturally occurring curcuminoids and related compounds against mosquito larvae.

The three curcuminoid components commonly isolated from Curcuma longa, curcumin (1), demethoxycurcumin (2), and bis-demethoxycurcumin (3) were separated and isolated from a commercially available turmeric extract product in high purity and sufficient amounts. Three more derivatives of curcumin, the di-O-demethylcurcumin (4), di-O-methylcurcumin (5) and the di-O-acetylcurcumin (6) were also synthesized and characterized. All six compounds were evaluated for their larvicidal effect against the mosquito Culex pipiens. Curcumin (1) exhibited highly potent larvicidal activity with LC(50) value of 19.07mgL(-1). Moreover, di-O-demethylcurcumin (4), was found to be equally active with LC(50) value of 12.42mgL(-1). Based on the LC(90) values of the two compounds, di-O-demethylcurcumin (4) was the most active of all, resulting in an LC(90) value of 29.40mgL(-1), almost half of the LC(90) value 61.63mgL(-1) found for compound 1. The rest of the compounds were inactive at concentrations even as high as 150mgL(-1) indicating a dependence of the larvicidal activity upon the substitution patent and the presence of aromatic hydroxyl and methoxy moieties. These results show for the first time the potential of this valuable natural product regarding its use as vector control agent.

Essential oils of indigenous in Greece six Juniperus taxa: chemical composition and larvicidal activity against the West Nile virus vector Culex pipiens.

The chemical composition of 14 essential oils (EOs), obtained from various parts (leaves, fruits, wood) of the six indigenous in Greece Juniperus family taxa, was determined by GC and GC/MS analysis. The insecticidal properties of these EOs were evaluated against Culex pipiens L. larvae of 3rd and early 4th instars, in order to delineate the relationship between the phytochemical content of the EOs and their larvicidal activities. The analytical data indicated that the EOs mainly consisted of monoterpenes, mostly cyclic and only occasionally aliphatic, and to a lesser percent, of diterpenes. The larvicidal bioassays against C. pipiens larvae revealed that the most active EO was derived from the wood of Juniperus drupacea and contains mainly non-oxygenated monoterpenes and a significant amount of diterpenes, displaying the highest chemodiversity. Its initial LC(50) value was 26.47 mg L(-1). On the contrary, the EO isolated from J. phoenicea berries, which consisted of monoterpenes (non-oxygenated, cyclic), was the less active displaying an LC(50) value of 96.69 mg L(-1). In respect to the contained phytochemicals, myrcene was assayed as the most toxic, displaying an LC(50) value of 33.83 mg L(-1), while the four isomers of pinene abundant in all EOs were less active exhibiting LC(50) values ranging from 70.40 to 94.88 mg L(-1). Results herein reveal that the EOs isolated from the studied Juniperus family taxa represent an inexpensive source of natural mosquito control mixtures.

Chemical composition and larvicidal activity of essential oils from six Apiaceae family taxa against the West Nile virus vector Culex pipiens.

The chemical compositions of essential oils (EOs) obtained from six different taxa of the Apiaceae family, Apioideae subfamily, belonging to three tribes and six different genuses were determined by gas chromatography and gas chromatography-mass spectrometry analyses. All examined plants grow naturally in different habitats throughout Greece. The insecticidal properties of these EOs were evaluated against Culex pipiens L. larvae of third and early fourth instars in order to delineate the relationship between the EOs phytochemical content and larvicidal activity. The analytical data indicated that the EOs are mainly consisted of monoterpenes, mostly cyclic and only occasionally aliphatic. The larvicidal bioassay results indicated that the oil of Oenanthe pimpinelloides L., which contains mainly nonoxygenated monoterpenes, possesses the highest activity against Cx. pipiens larvae, displaying a LC(50) value of 40.26 mg/L. On the contrary, the EO of Elaeoselinum asclepium (L.) Bertol, which is consisted of pinenes and oxygenated monoterpenes, was the less active (LC(50) value of 96.96 mg/L). These results reveal that the nonoxygenated monoterpenes possess potent insecticidal activities against Cx. pipiens L. and the EO of O. pimpinelloides L. represents an inexpensive source of natural pest control mixture.

Volatile secondary metabolite pattern of callus cultures of Chamomilla recutita.

The effects of various plant growth regulators and culture conditions on the production of volatile secondary metabolites from callus cultures of Chamomile (Chamomilla recutita) inflorescence were investigated and the most efficient conditions were determined. The essential oil composition was assayed by GC-MS analysis and found to contain chamomillol, gossonorol, cubenol, alpha-cadinol, (-)-alpha-bisabolol, 1-azulenethanol acetate and (-)-alpha-bisabolol acetate.