Préparation à l&#39;avance de la micro-injection: <ol><li> Moustiques nourrissent de sang: pour l&#39;injection du lundi au mercredi femelles se nourrissent le vendredi précédent. Pour injecter jeudi et vendredi, les femelles se nourrissent le lundi de la même semaine. </li><li> Préparer les aiguilles à quartz utilisant le programme 2 et tampon isotonique (voir matériaux). </li><li> &quot;Pose du tube&quot; pour les embryons est le flacon de culture régulière drosophile. Coton humide dans le fond, avec un disque de papier filtre humide qui la recouvre. </li><li> Préparer lamelle en plastique en collant double-face de la cassette à une extrémité. Bande de garniture à lamelle afin qu&#39;elle se termine au bord de lamelle. </li><li> Préparer l&#39;huile pour la dessiccation. </li><li> Préparez une boîte de Pétri avec tampon isotonique de transférer les embryons à couver. </li></ol> Mise en place de la pose forcée: <ol><li> Recueillir le sang de 60 à 10 femelles nourries avec l&#39;utilisation d&#39;un aspirateur et de les transférer dans le flacon de culture de drosophiles avec du coton et du papier filtre humide avec tampon isotonique. </li><li> Les moustiques sont ensuite remis en conditions insectery dans l&#39;obscurité et autorisés à pondre des oeufs pendant 1 h et 15 min. </li><li> Laissez adultes volent dans la cage et retirez le disque de papier filtre avec des embryons. </li><li> Pour aligner les œufs, le faire sous la portée de dissection. Recueillir des grappes d&#39;oeufs avec un pinceau fin (Sable, No 0000) et le transfert à un carré préparés de papier Whatman 3MM imbibé de tampon isotonique. Conservez le papier tous les temps humides. Ne laissez pas les œufs se dessèchent. Retirez le exochorion avec le pinceau, il aide à mieux coller les oeufs sur la bande. </li><li> Utiliser fines pince n ° 5 ou un pinceau fin, ramasser embryons gris foncé et d&#39;organiser en ligne sur le carré du papier Whatman 3MM humide avec un tampon isotonique. Line up de 20 à 30 embryons. Tous les embryons doivent être dans la même orientation que l&#39;injection doit être au pôle postérieur. L&#39;extrémité antérieure de l&#39;embryon est légèrement plus large que le postérieur. Puis, en utilisant des bandes de papier Whatman 3MM, sécher le filtre où les œufs sont alignés en appuyant fort sur les deux côtés de la ligne d&#39;oeuf, ne pas toucher les oeufs. </li><li> Pour transférer les œufs, inverser la diapositive contenant le ruban adhésif double face (Médecine), et appuyez doucement contre les oeufs. L&#39;extrémité postérieure des œufs doivent être très proches du bord de l&#39;adhésif double face. L&#39;humidité du papier est essentiel pour cela, si elle est trop humide, ils ne collent pas. Je ne la dessiccation des embryons de la deuxième 5-10, mais le temps de dessiccation dépend de l&#39;humide et la température dans la chambre de la micro-injection. </li><li> La dessiccation est cruciale pour les oeufs: peu trop et les embryons ne seront pas éclore. Sans l&#39;ADN de dessiccation n&#39;est pas d&#39;entrer dans l&#39;embryon. </li><li> Couverture embryons desséchés avec de l&#39;huile d&#39;halocarbures pour éviter la dessiccation plus loin. </li></ol> Microinjection des embryons: L&#39;aspect le plus important de l&#39;injection est la qualité de l&#39;aiguille (voir note). <ol><li> Remplir une seringue avec la solution d&#39;ADN à injecter en utilisant un microloader (Eppendorf # 5242 956,003). Très peu d&#39;injection soliution est nécessaire, 1 à 2 ml. </li><li> Connectez l&#39;aiguille à la Transjector Eppendorf, qui contrôle le temps d&#39;injection et de pression, ainsi que la contre-pression. La microinjection est réalisée à l&#39;aide d&#39;un microscope avec une scène en mouvement (Leica) à un grossissement x 10 et micromanipulateur (Leica). Si nécessaire, une platine de microscope soulevées peuvent être préparés par l&#39;empilement de lames de microscope 4-5. Placer la lamelle transportant des embryons sur scène surélevée. Les embryons sont injectés à un angle de 150 °; pénétration doit être au pôle postérieur. Pour l&#39;injection, je garde l&#39;aiguille immobile et déplacer le platine du microscope. Lorsque l&#39;aiguille est nouveau, la pointe est scellée et les pauses habituellement dans la première injection. Je travaille toujours sur la pression nécessaire pour expulser une petite goutte d&#39;ADN dans l&#39;huile entre chaque injection. </li><li> J&#39;ai mis du temps d&#39;injection de 0,2 à 0,9 sec et 1000 hPa lorsque l&#39;aiguille est nouveau. Je varie la pression et le temps jusqu&#39;à ce qu&#39;une gouttelette est petite vu sortir de l&#39;aiguille dans l&#39;huile. La contre-pression doit être réglée à environ 100. Comme l&#39;aiguille s&#39;use, la pression d&#39;injection doit être réduite pour garder le faible volume d&#39;injection (environ 300 hPa). Après cela, la pointe de l&#39;aiguille est trop grande et tue la plupart des embryons de l&#39;. </li><li> Après l&#39;injection, enlever l&#39;huile avec du papier absorbant et recouvrir la lame avec les embryons et les placer dans la boîte de Pétri avec le tampon isotonique. Placez les plats dans le insectory et les y laisser jusqu&#39;à ce que les embryons éclosent. Ils commencent à éclore après 2-6 jours. Transfert aux larves de l&#39;eau distillée avec la pincée de nourriture pour poissons sol. </li></ol> Remarques L&#39;ADN pour l&#39;injection: la solution pour injection d&#39;ADN plasmidique a été isolé en utilisant Endofree kit plasmide.Construire le plasmide et mélanger plasmide assistant ensemble dans la bonne concentration, précipité à l&#39;isopropanol. Le culot a été lavé avec de l&#39;éthanol 70% avant la remise en suspension dans le tampon d&#39;injection. Avant l&#39;injection, l&#39;ADN propre à l&#39;aide Millex-GV colonne. Buffer Izotonic: <table border="1" cellpadding="8"><tbody><tr><td> 1,68 M de NaCl - 88,3 ml 1,68 M de NaCl - 2,9 m 1 M HEPES -10,7 ml 1,12 M CaCl 2 à 2,2 ml dH 2 O - 896 ml </td><td> OU </td><td> NaCl 5 M - 29,67 ml 1 M KCl - 4,87 ml 1 Hepes M - 10,7 ml 1,12 M CaCl 2 à 2,2 ml dH 2 O - 952,56 ml </td></tr></tbody></table> Ajuster à pH 7,2

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Injection Quartz needles</td>
<td>&nbsp;</td>
<td>Sutter Instrument</td>
<td>&nbsp;</td>
<td>O.D.- 1.0 mm, I.D.-0.70, 10 cm length, are prepared by drawing capillary tubing into a fine 3-8 um with a shaft of approximately 100-300 mm micropipette puller. </td>
</tr>
<tr>
<td>Quartz puller</td>
<td>Tool</td>
<td>Sutter Instrument Co</td>
<td>P-2000</td>
<td>Set-up: Heat-275, Fil-3, Vel-38, Del-250, Pull-141</td>
</tr>
<tr>
<td>EndoFree plasmid kit</td>
<td>&nbsp;</td>
<td>Quiagen</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Isopropanol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Millex-GV column</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>SLGV RO4 NL</td>
<td>for DNA cleaning</td>
</tr>
<tr>
<td>Construct DNA</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.5 mg/ml </td>
</tr>
<tr>
<td>Helper plasmid</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.3 mg/ml </td>
</tr>
<tr>
<td>Injection solution</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>1x solution: 5mM KCl, 0.1mM sodium phosphate, ph 6. 8</td>
</tr>
<tr>
<td>Isotonic buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>see recipe in the protocol section</td>
</tr>
<tr>
<td>Dessication oil</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Halocarbon oil 700:Halocarbon oil 27(1:1). Mix well. Prepare in advance of microinjection.</td>
</tr>
<tr>
<td>Blood</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>to feed mosquitos</td>
</tr>
<tr>
<td>Anopheles stephensi</td>
<td>Animal</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Mosquitos, male and female.</td>
</tr>
<tr>
<td>Drosophila culture vial</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>for egg laying</td>
</tr>
<tr>
<td>Petri dishes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>with isotonic buffer, for embryo hatching</td>
</tr>
<tr>
<td>Fine paintbrush</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Sable # 0000</td>
</tr>
<tr>
<td>3MM Whatman paper</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Forceps #5</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>double sided tape</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microloader</td>
<td>&nbsp;</td>
<td>Eppendorf</td>
<td>5242 956.003</td>
<td>&nbsp;</td>
<tr>
<td>Transjector</td>
<td>&nbsp;</td>
<td>Eppendorf</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microscope</td>
<td>&nbsp;</td>
<td>Leica</td>
<td>&nbsp;</td>
<td>with moving stage and micromanipulator, set at 10x magnification </td>
</tr>		</tbody>
		</table>
		</div>
			<div>
					</div>

injection of an. stephensi embryos to generate malaria-resistant mosquitoes

L&#39;introduction de gènes exogènes dans les génomes des moustiques nécessite des techniques de micro-injection adaptés aux espèces d&#39;intérêt particulier. Ce protocole vidéo montre une méthode utilisée par le laboratoire de James microinject constructions d&#39;ADN dans des embryons stephensi Anopheles pour la génération de moustiques transformé. Les techniques de préparation des aiguilles de microinjection, la collecte et la préparation des embryons et l&#39;exécution des micro-injection sont illustrés.

Watch this Scientific Journal Video about Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes at JoVE.com

Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes

Moustiques Anopheles stephensi sont des vecteurs de la malaria qui habitent en Inde et en Asie. Cette vidéo montre la technique pour effectuer des micro-injections de cette espèce avec des transgènes qui confèrent une résistance à la malaria au moustique. Une grande partie de la méthodologie démontrée dans cette vidéo est applicable à des techniques de micro-injection d&#39;autres espèces de moustiques.

Injection d&#39;un. Embryons stephensi pour générer le paludisme résistant à moustiques

The introduction of exogenous genes into the genomes of mosquitoes requires microinjection techniques tailored to the specific species of interest. This ...

injection-an-stephensi-embryos-to-generate-malaria-resistant

Research

JoVE Journal

Biology

12.9K vues.  University of California, Irvine (UCI). Moustiques Anopheles stephensi sont des vecteurs de la malaria qui habitent en Inde et en Asie. Cette vidéo montre la technique pour effectuer des micro-injections de cette espèce avec des transgènes qui confèrent une résistance à la malaria au moustique. Une grande partie de la méthodologie démontrée dans cette vidéo est applicable à des techniques de micro-injection d'autres espèces de moustiques.

Vidéo: Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes

Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes

In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue fever.  The techniques for correctly preparing microinjection needles, dessicating embryos, and performing microinjection are demonstrated.

In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue fever. The techniques for correctly preparing microinjection needles, dessicating embryos, and performing microinjection are demonstrated.

Microinjection d&#39;embryons aegypti a. Pour obtenir les moustiques transgéniques

In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue ...

Recherche

Biologie

In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

In-utero in-vivo injection and electroporation of the embryonic rat neocortex provides a powerful tool for the manipulation of individual progenitors lining the walls of the lateral ventricle. This technique is now widely used to study the processes involved in corticogenesis by over-expressing or knocking down genes and observing the effects on cellular proliferation, migration, and differentiation. In comparison to traditional knockout strategies, in-utero electroporation provides a rapid means to manipulate a population of cells during a specific temporal window. In this video protocol, we outline the experimental methodology for preparing rats for surgery, exposing the uterine horns through laporatomy, injecting DNA into the lateral ventricles of the developing embryo, electroporating DNA into the progenitors lining the lateral wall, and caring for animals post-surgery. Our laboratory uses this protocol for surgeries on E15-E21 rats, however it is most commonly performed at E16 as shown in this video.

En injection intraventriculaire Utero et l&#39;électroporation d&#39;embryons de rat E16

In-utero in-vivo injection and electroporation of the embryonic rat neocortex provides a powerful tool for the manipulation of individual progenitors ...

In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos

In-utero in-vivo injection and electroporation of the embryonic mouse neocortex provides a powerful tool for the manipulation of individual progenitors lining the walls of the lateral ventricle. This technique is now widely used to study the processes involved in corticogenesis by over-expressing or knocking down genes and observing the effects on cellular proliferation, migration, and differentiation. In comparison to traditional knockout strategies, in-utero electroporation provides a rapid means to manipulate a population of cells during a specific temporal window. In this video protocol we outline the experimental methodology for preparing mice for surgery, exposing the uterine horns through laporatomy, injecting DNA into the lateral ventricles of the developing embryo, electroporating DNA into the progenitors lining the lateral wall, and caring for animals post-surgery. Our laboratory uses this protocol for surgeries on E13-E16 mice, however, it is most commonly performed at E15, as shown in this video. 

En injection intraventriculaire Utero et l&#39;électroporation d&#39;embryons de souris E15

In-utero in-vivo injection and electroporation of the embryonic mouse neocortex provides a powerful tool for the manipulation of individual progenitors ...

Microinjection of Zebrafish Embryos to Analyze Gene Function

One of the advantages of studying zebrafish is the ease and speed of manipulating protein levels in the embryo.  Morpholinos, which are synthetic oligonucleotides with antisense complementarity to target RNAs, can be added to the embryo to reduce the expression of a particular gene product.  Conversely, processed mRNA can be added to the embryo to increase levels of a gene product.  The vehicle for adding either mRNA or morpholino to an embryo is microinjection.  Microinjection is efficient and rapid, allowing for the injection of hundreds of embryos per hour.  This video shows all the steps involved in microinjection.  Briefly, eggs are collected immediately after being laid and lined up against a microscope slide in a Petri dish.  Next, a fine-tipped needle loaded with injection material is connected to a microinjector and an air source, and the microinjector controls are adjusted to produce a desirable injection volume.  Finally, the needle is plunged into the embryo's yolk and the morpholino or mRNA is expelled.

This video shows how morpholino or mRNA can be injected into zebrafish embryos at the one-cell stage to decrease or increase the level of specific gene products during subsequent development.

Microinjection d&#39;embryons de poissons zèbres pour analyser la fonction génique

One of the advantages of studying zebrafish is the ease and speed of manipulating protein levels in the embryo.  Morpholinos, which are synthetic ...

RNAi Interference by dsRNA Injection into Drosophila Embryos

Genetic screening is one of the most powerful methods available for gaining insights into complex biological process 1. Over the years many improvements and tools for genetic manipulation have become available in Drosophila 2. Soon after the initial discovery by Frie and Mello 3 that double stranded RNA can be used to knockdown the activity of individual genes in Caenorhabditis elegans, RNA interference (RNAi) was shown to provide a powerful reverse genetic approach to analyze gene functions in Drosophila organ development 4, 5.
Many organs, including lung, kidney, liver, and vascular system, are composed of branched tubular networks that transport vital fluids or gases 6, 7. The analysis of Drosophila tracheal formation provides an excellent model system to study the morphogenesis of other tubular organs 8. The Berkeley Drosophila genome project has revealed hundreds of genes that are expressed in the tracheal system. To study the molecular and cellular mechanism of tube formation, the challenge is to understand the roles of these genes in tracheal development. Here, we described a detailed method of dsRNA injection into Drosophila embryo to knockdown individual gene expression. We successfully knocked down endogenous dysfusion(dys) gene expression by dsRNA injection. Dys is a bHLH-PAS protein expressed in tracheal fusion cells, and it is required for tracheal branch fusion 9, 10. dys-RNAi completely eliminated dys expression and resulted in tracheal fusion defect. This relatively simple method provides a tool to identify genes requried for tissure and organ development in Drosophila.

RNAi Interference by dsRNA Injection into Drosophila Embryos

RNA interference has been proven very effective to analyze gene function in Drosophila tracheal development. A detailed protocol used by Jiang lab to inject dsRNA into fly embryos to knockdown gene expression is illustrated. This technique has the potential for screening genes required for tissue and organ development in Drosophila.

Interférences ARNi par injection dans les ARNdb Drosophile Embryons

Genetic screening is one of the most powerful methods available for gaining insights into complex biological process 1. Over the years many improvements ...

An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos

Many cellular processes are controlled by multisubunit protein complexes. Frequently these complexes form transiently and require native environment to assemble. Therefore, to identify these functional protein complexes, it is important to stabilize them in vivo before cell lysis and subsequent purification. Here we describe a method used to isolate large bona fide protein complexes from Drosophila embryos. This method is based on embryo permeabilization and stabilization of the complexes inside the embryos by in vivo crosslinking using a low concentration of formaldehyde, which can easily cross the cell membrane. Subsequently, the protein complex of interest is immunopurified followed by gel purification and analyzed by mass spectrometry. We illustrate this method using purification of a Tudor protein complex, which is essential for germline development. Tudor is a large protein, which contains multiple Tudor domains - small modules that interact with methylated arginines or lysines of target proteins. This method can be adapted for isolation of native protein complexes from different organisms and tissues.

An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos

Multi-component protein complexes play crucial roles during cellular function and development. Here we describe a method used to isolate native protein complexes from Drosophila embryos after in vivo crosslinking followed by purification of the crosslinked complexes for subsequent structure-function analysis.

Un In vivo Approche de réticulation pour isoler les complexes protéiques De Drosophila Embryons

Many cellular processes are controlled by multisubunit protein complexes. Frequently these complexes form transiently and require native environment to ...

Toxicological Assays for Testing Effects of an Epigenetic Drug on Development, Fecundity and Survivorship of Malaria Mosquitoes

Insecticidal resistance poses a major problem for malaria control programs. Mosquitoes adapt to a wide range of changes in the environment quickly, making malaria control an omnipresent problem in tropical countries. The emergence of insecticide resistant populations warrants the exploration of novel drug target pathways and compounds for vector mosquito control. Epigenetic drugs are well established in cancer research, however not much is known about their effects on insects. This study provides a simple protocol for examining the toxicological effects of 3-Deazaneplanocin A (DZNep), an experimental epigenetic drug for cancer therapy, on the malaria vector, Anopheles gambiae. A concentration-dependent increase in mortality and decrease in size was observed in immature mosquitoes exposed to DZNep, whereas the compound reduced the fecundity of adult mosquitoes relative to control treatments. In addition, there was a drug-dependent decrease in S-adenosylhomocysteine (SAH) hydrolase activity in mosquitoes following exposure to DZNep relative to control treatments. These protocols provide the researcher with a simple, step-by-step procedure to assess multiple toxicological endpoints for an experimental drug and, in turn, demonstrate a unique multi-prong approach for exploring the toxicological effects of water-soluble epigenetic drugs or compounds of interest against vector mosquitoes and other insects.

A protocol is developed to examine the effects of an epigenetic drug DZNep on the development, fecundity and survivorship of mosquitoes. Here we describe procedures for the aqueous exposure of DZNep to immature mosquitoes and a blood-based exposure of DZNep to adult mosquitoes in addition to measuring SAH hydrolase inhibition.

Les analyses toxicologiques pour Effets d&#39;essais d&#39;un médicament épigénétique sur le développement, la fécondité et de la survie du paludisme moustiques

Insecticidal resistance poses a major problem for malaria control programs. Mosquitoes adapt to a wide range of changes in the environment quickly, making ...

Protocols for Testing the Toxicity of Novel Insecticidal Chemistries to Mosquitoes

New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such as Zika, dengue and malaria. Assays for rapid, high-throughput analyses of unformulated novel chemistries against mosquito larvae and adults are presented. We describe protocols for single point-dose and dose response assays to evaluate the toxicity of small molecule chemistries to the Aedes aegypti vector of Zika, dengue and yellow fever,&#160;the malaria vector, Anopheles gambiae and the northern house mosquito, Culex quinquefasciatus, on contact and via ingestion. As an example, we evaluated the toxicity of amitriptyline, a small molecule antagonist of G protein-coupled receptors, via larval, adult topical and adult blood-feeding assay. The protocols provide a starting point to investigate insecticide potential. Results are discussed in the context of additional experiments to explore product applications and mechanisms for delivery.

Protocols are described for assessing the toxicity of chemistries to immature and adult mosquitoes for development as new classes of larvicides, adulticides and endectocides. The protocols enable high-throughput testing of multiple chemistries at single-point dose and subsequent evaluation via dose response assay to determine toxicity on contact or via ingestion.

Protocoles pour tester la toxicité des nouvelles chimies insecticides aux moustiques

New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such ...

Forced Salivation As a Method to Analyze Vector Competence of Mosquitoes

Vector competence is defined as the potential of a mosquito species to transmit a mosquito-borne virus (mobovirus) to a vertebrate host. Viable virus particles are transmitted during a blood meal via the saliva of an infected mosquito. Forced salivation assays allow determining the vector potential on the basis of single mosquitoes, avoiding the use of animal experiments. The method is suitable to analyze a large number of mosquitoes in one experiment within a short period of time. Forced salivation assays were used to analyze 856 individual mosquitoes trapped in Germany, including two different Culex pipiens pipiens biotypes, Culex torrentium as well as Aedes albopictus, which were experimentally infected with Zika virus (ZIKV) and incubated at 18 &#176;C or 27 &#176;C for two and three weeks. The results indicated the lack of vector competence of the different Culex taxa for ZIKV. In contrast, Aedes albopictus was susceptible to ZIKV, but only at 27 &#176;C, with transmission rates similar to an Aedes aegypti laboratory colony tested in parallel.

For efficient control of mosquito borne virus transmission, the knowledge of the vector potential of respective mosquitoes is of particular interest. We describe forced salivation as a method to analyze vector competence of Aedes albopictus and three different Culex taxa for the transmission of Zika virus.

Forcé de Salivation comme une méthode pour analyser la compétence vectorielle de moustiques

Vector competence is defined as the potential of a mosquito species to transmit a mosquito-borne virus (mobovirus) to a vertebrate host. Viable virus ...

A Blood-Free Diet to Rear Anopheline Mosquitoes

Malaria research requires large-scale breeding and production conditions for mosquitoes (Anopheles spp.) in captivity. The sustainable and reliable production of mosquitoes is currently inhibited by the supply of fresh vertebrate blood. Alternatives to blood are required to promote efficient control strategies for malaria and other vector borne diseases that are transmitted by blood feeding insects. With this in mind, artificial liquid diets were formulated as substitutes for fresh vertebrate blood. Herein we report a blood-free artificial liquid diet that delivers feeding rates similar to blood and mimics the physiological effects of a fresh vertebrate blood meal. The diet induces ovarian and egg maturation of Anopheles mosquitoes and also produces good larval survival and development of functional adults. The formulated blood-free liquid diet is an important advance towards sustainable mosquito breeding in captivity and will reduce the maintenance costs of mosquito colonies and eliminate the need for fresh vertebrate blood.

A protocol is presented for formulation of a blood-free artificial diet to feed Anopheles mosquitoes in captivity. This diet has a similar performance to vertebrate blood and triggers oogenesis and egg maturation and produces viable adult progeny.

Un régime sans sang pour les moustiques anophèles arrière

Malaria research requires large-scale breeding and production conditions for mosquitoes (Anopheles spp.) in captivity. The sustainable and reliable ...