trichogramma dendrolimi artificial host interaction and venom extraction

Parassita artificiale dell'ospite mediante Trichogramma dendrolimi ed estrazione del veleno

Parasitoid wasps are parasitic hymenopteran insects that are important resources for biological control1. There is a wide variety of parasitoid wasps, with over 600,000 estimated species2. The diversity of parasitoid wasps far exceeds that of other venomous arthropods, such as snakes, cone snails, spiders, scorpions, and bees. Venom is an important parasitic factor in parasitoid wasps. For successful parasitism, venom is injected into the host, modulating the host&#39;s behavior, immunity, development, and metabolism3. Moreover, the venom of parasitoid wasps displays remarkable diversity in its molecular structures, targets, and functions, reflecting complex coevolution with their hosts. Thus, parasitoid venom is a valuable and underappreciated resource of active molecules for insecticidal or medical purposes4. Unlike the venom of snakes, cone snails, spiders, scorpions, and bees, parasitoid wasp venom cannot be collected by direct stimulation or electrical stimulation5. The current method of extraction of parasitoid wasp venom is to dissect the venom reservoir. However, parasitoid wasps are often small, and dissection of parasitoid wasps requires high technical skills. Therefore, if we can find a way to collect the venom of parasitoid wasps efficiently and conveniently, it will be of great help to research the venom of parasitoid wasps.
Trichogramma (Hymenoptera: Trichogrammatidae) is a genus of tiny (~0.5 mm long) parasitoid wasps6. These wasps are among the most widely used biocontrol agents, particularly targeting eggs of various lepidopteran pests in both agriculture and forests. For example, T. dendrolimi, one of the most widely used Trichogramma species in China, has been extensively applied to control a variety of agricultural and forestry pests, such as Dendrolimus superans, Ostrinia furnacalis, and Chilo suppressalis. Previous studies showed that Trichogramma wasps could inject their eggs into artificial hosts7. Artificial hosts can be created using materials such as wax8, agar9, Parafilm10, and plastic film11. The solution in artificial hosts that induces sufficient oviposition for Trichogramma can be simple, such as amino acids or inorganic salts12. Based on the characteristic that T. dendrolimi can parasitize artificial hosts, this study provides a new method for extracting venom from parasitoid wasps using artificial hosts. This approach aims to address the shortcomings of low yield, low purity, and susceptibility to contamination in current extraction techniques. By using this method, a large amount of high-purity venom from T. dendrolimi can be extracted, which meets the needs of scientific research and screening of bioactive molecules for insecticidal or medical purposes.

Autore in primo piano: Metodo efficiente di estrazione del veleno dalle vespe parassitoidi Trichogramma 

Estrazione del veleno dalla vespa parassitoide Trichogramma dendrolimi utilizzando un ospite artificiale

The lack of efficient venom extraction methods often limits the research on the venom of parasitoid wasps, especially in tiny parasitoid wasps. This study provides an efficient method to extract the venom of Trichogramma with follow up post study of Trichogramma venom, such as the protein compensation and the venom function analysis. Previously, no methods of exchanging of parasitoid wasp venom requires the distinguishing of venom reserve.However, parasitoid wasps are tiny. Not only are the technical requirements of distinguishing venom was high, but continuation of other tissues during dissection is also common. Our normal method uses artificial host, thus avoiding such problems.In the future, we will focus on the agriculture and the medical applications of Trichogramma venom. We aim to utilize specific insecticidal venom genes for pest control. Meanwhile, we plan to investigate the untapped potential of Trichogramma venom in methodology, particularly its prospective utility in enhancing immunotherapy techniques, and in the discovery of novel antibiotic molecules.

Watch this Scientific Journal Video about Trichogramma dendrolimi  Artificial Host Interaction and Venom Extraction at JoVE.com

Trichogramma dendrolimi  Artificial Host Interaction and Venom Extraction  

Dendrolimi di Trichogramma Interazione con l'ospite artificiale ed estrazione del veleno

Per iniziare, prendi una pellicola di plastica in polietilene lunga 16 centimetri, larga 12 centimetri e spessa 20 micrometri. Utilizzare uno strumento di macinazione del vetro per premere 30 sporgenze semicircolari in un layout PCR standard a 96 pozzetti. Esporre entrambi i lati del film plastico di polietilene pressato alla luce UV per un'ora per disinfettarlo.Aggiungere una piccola quantità di alcol polivinilico al 10% sulla superficie semicircolare. Successivamente, metti circa 3.000 vespe trichogramma dendrolimi femmine anestetizzate in una scatola di raccolta. Posizionare il lato convesso della carta uovo in pellicola verso il contenitore di raccolta e fissare i bordi con un elastico.Ora aggiungi quattro microlitri di soluzione amminoacidica a ciascuna sporgenza. Quindi coprilo con un altro pezzo di pellicola di plastica in polietilene. Utilizzare un elastico per coprire ermeticamente il contenitore di raccolta con due fogli di plastica.Lascia che le vespe parassitino liberamente per quattro-otto ore. Fornire acqua di saccarosio al 10% attraverso il cotone bagnato durante la parassitizzazione. Successivamente, ottenere la soluzione di amminoacidi parassitati dalla sporgenza interna della carta uovo artificiale e trasferirla nel tappo di provette da 1,45 millilitri.Coprire il tappo del tubo con una rete di nylon da 10 micrometri, avente un diametro di 25 millimetri. Quindi fissare leggermente la rete di nylon nel tubo. Centrifugare la provetta in alto a destra a 1, 360 g per 10 secondi.Raccogli la soluzione filtrata contenente il veleno. Conservare il veleno a meno 80 gradi Celsius per ulteriori analisi. L'analisi BCA dei campioni di veleno ha mostrato che la concentrazione di proteine del veleno variava da 0,35 a 0,46 microgrammi per microlitro.Il controllo negativo della soluzione di amminoacidi aveva solo una concentrazione proteica compresa tra 0,03 e 0,05 microgrammi per microlitro. L'analisi SDS-PAGE del veleno ha mostrato una gamma di proteine che va da meno di 10 kilodalton a oltre 130 kilodalton.

Research

JoVE Journal

Biochemistry

Trichogramma dendrolimi Artificial Host Interaction and Venom Extraction

Extracting Venom from the Parasitoid Wasp Trichogramma dendrolimi Using an Artificial Host

Parasitoid wasps are a diverse group of hymenopteran insects that serve as invaluable resources for pest biocontrol. To ensure successful parasitism, ...