Mikroenjeksiyon, önceden hazırlanması: <ol><li> Kan linki sivrisinek: Pazartesi - Çarşamba önceki Cuma günü linki kadın enjeksiyon için. Aynı hafta Pazartesi, Perşembe ve Cuma günleri, yem kadın enjekte etmek. </li><li> Kuvars iğne programı 2 ve izotonik tampon (Malzeme) kullanarak hazırlayın. </li><li> Embriyolar &quot;tüp Döşeme&quot; düzenli Drosophila kültür flakon. Kapsayan filtre kağıdı ıslak bir disk ile alt Islak pamuk. </li><li> Bir ucunda çift taraflı bant yapıştırma, plastik kapak kayma hazırlayın. Trim kaset kapağı için, kapak kayma kenarında biter, böylece kayma. </li><li> Kuruma petrol hazırlayın. </li><li> Petri kabı, kuluçka için embriyo transferine izotonik tamponu ile hazırlayın. </li></ol> Zorla döşeme ayarlayın: <ol><li> Toplayın ve bir aspiratör ile 6-10 kan beslenen kadınlarda, pamuk ve izotonik tampon ıslak filtre kağıdı ile Drosophila kültür flakon aktarabilirsiniz . </li><li> Sivrisinek sonra karanlıkta insectery koşullar içine geri koyun ve 1 saat ve 15 dakika için yumurtalarını bırakmak için izin. </li><li> Yetişkin kafes içine sinek ve embriyolar ile filtre kağıdı diski çıkarın. </li><li> Yumurta kadar satır için, diseksiyon kapsamında bunu. (Sable, No 0000) ince bir fırça ile yumurta demet toplayın ve hazırlanmış bir kare izotonik tampon ile ıslatılmış 3MM whatman kağıt transfer. Tüm zamanların ıslak kağıt tutun. Yumurta kurutulmuş almak izin vermeyin. Exochorion fırça ile kaldırın, daha iyi bant yumurta sopa yardımcı olur. </li><li> Ince forsepsle No 5 ya da ince bir fırça kullanarak, koyu gri embriyolar pick up ve izotonik tampon ile ıslak 3MM whatman kare doğrultusunda düzenlemek. 20-30 embriyolardan Hattı. Enjeksiyon arka kutupta yer gibi tüm embriyolar aynı yönde olmalıdır. Embriyoların ön arka biraz daha geniş. Sonra 3MM whatman kağıt şeritleri kullanarak, yumurta, yumurta hattının her iki tarafında sert basarak, yumurta dokunmadan dizilmiş filtre kurulayın. </li><li> Yumurta transfer etmek için, çift taraflı bant (Tıp) içeren slayt, invert ve yumurta karşı hafifçe basın. Yumurta arka ucunda çift taraflı bant kenarına çok yakın olması. Yapışmaz çok ıslak ise kağıt ıslaklık, bunun için kritik öneme sahiptir. 5-10 saniyelik embriyoların kuruma, ancak kuruma süresi mikroenjeksiyon oda nemli ve sıcaklık bağlıdır. </li><li> Biraz çok ve embriyolar yumurtadan olmaz: Kuruma yumurta için çok önemlidir. Kuruma DNA olmadan embriyo girecek değildir. </li><li> Kurutulmuş embriyolar daha fazla kuruma önlemek için Halokarbon yağ ile kaplayın. </li></ol> Embriyoların Mikroenjeksiyon: En önemli özelliği iyi bir enjeksiyon (Not bakınız) iğne kalitesidir. <ol><li> Microloader (Eppendorf # 5242 956,003) kullanarak enjekte edilecek DNA çözümü ile bir iğne doldurun. Çok az enjeksiyon soliution, 1 - 2 ml gereklidir. </li><li> Iğne, enjeksiyon süresi ve basınç kontrolleri Eppendorf Transjector, yanı sıra backpressure bağlayın. Mikroenjeksiyon x 10 büyütme ve mikromanipülatör (Leica) hareketli bir sahne (Leica) bir mikroskop kullanılarak yapılır. Gerekirse, yükseltilmiş bir mikroskop aşamada 4-5 mikroskop lamı istifleme tarafından hazırlanmış olabilir. Yükseltilmiş sahneye taşıyan embriyolar kapak kayma yerleştirin. Embriyolar 150 ° açıyla enjekte edilir; penetrasyon arka kutupta olmalıdır. Enjeksiyon için iğne sabit tutmak ve mikroskop aşamasına taşımak. Iğne ucu kapalı ve genellikle ilk enjeksiyondan tatili. Ben her zaman, her enjeksiyon arasında DNA&#39;nın küçük bir damla petrol sınırdışı için gerekli olan basınç çalışır. </li><li> Ben 0.2 enjeksiyon süresi 0.9 saniye ve 1000 hPa iğne yeni. Kadar küçük bir damlacık yağın içine iğne çıkan basınç ve zaman değişir. Backpressure yaklaşık 100 ayarlanması gerekir. Iğne ucu aşınmış alır gibi, enjeksiyon basıncı, enjeksiyon hacmi düşük tutmak için (yaklaşık 300 hPa) azaltılmalıdır. Bundan sonra, iğne ucu çok büyük ve embriyoların çoğu öldürüyor. </li><li> Enjeksiyondan sonra, kağıt mendil ile petrol çıkarmak ve embriyo ve izotonik tampon Petri kabında yer slayt kapağı. Insectory yemekleri yerleştirin ve embriyoların yumurtadan kadar orada bırakın. Bunlar 2-6 gün sonra yumurtadan çıkar başlar. Tutam toprak balık yemi ile distile su larva transferi. </li></ol> Notlar Enjeksiyon için DNA: enjeksiyon için Plazmid DNA çözüm EndoFree plazmid kiti kullanılarak izole edilmiştir.Plazmid Construct ve doğru konsantrasyon birlikte Yardımcısı plazmid karışımı, isopropanol hızlandırdı. Pelet enjeksiyon tampon resuspending önce% 70 etanol ile yıkandı. Enjeksiyon, önce Millex-GV sütun kullanarak temiz DNA. Izotonic tampon: <table border="1" cellpadding="8"><tbody><tr><td> 1.68 M NaCl - 88.3 ml 1.68 M NaCl - 2.9 m 1 M -10.7 ml HEPES 1.12 M CaCl 2 - 2,2 ml dH 2 O - 896 ml </td><td> VEYA </td><td> 5 M NaCl - 29,67 ml 1 M KCI - 4.87 ml 1 M HEPES - 10.7 ml 1.12 M CaCl 2 - 2,2 ml dH 2 O - 952,56 ml </td></tr></tbody></table> PH 7.2 &#39;ye ayarlayın

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<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<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

Sivrisinek genomların ekzojen genlerin giriş, özel ilgi türü için hazırlanmış mikroenjeksiyon teknikleri gerektirir. Bu video protokol dönüştürülmüş sivrisineklerin nesil için Anofel stephensi embriyolar DNA yapıları microinject James laboratuvar tarafından kullanılan bir yöntem gösteriyor. Teknikleri, mikroenjeksiyon iğneler hazırlanması, toplanması ve hazırlanması embriyolar ve mikroenjeksiyon işlemi için gösterilmiştir.

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

Anofel stephensi sivrisinekler, sıtma yaşayan Hindistan ve Asya&#39;da vektörleri. Bu video bu türün sivrisinek sıtma karşı direnç kazandırırlar transgenlerin microinjections gerçekleştirmek için teknik göstermektedir. Bu video gösterdi metodoloji diğer sivrisinek türlerinin mikroenjeksiyon teknikleri uygulanabilir.

Bir, Enjeksiyon. stephensi Embriyolar Sıtma dayanıklı Sivrisinekler oluşturun

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 Görüntüleme.  University of California, Irvine (UCI). Anofel stephensi sivrisinekler, sıtma yaşayan Hindistan ve Asya'da vektörleri. Bu video bu türün sivrisinek sıtma karşı direnç kazandırırlar transgenlerin microinjections gerçekleştirmek için teknik göstermektedir. Bu video gösterdi metodoloji diğer sivrisinek türlerinin mikroenjeksiyon teknikleri uygulanabilir.

Video: 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.

Transgenik Sivrisinekler Edinme A. aegypti Embriyolar Mikroenjeksiyon

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

Biyoloji

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.

E16 Rat Embriyolar Utero İntraventriküler Enjeksiyon ve Elektroporasyon

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. 

E15 Fare Embriyolar Utero İntraventriküler Enjeksiyon ve Elektroporasyon

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.

Gen Fonksiyon Analiz Zebra balığı Embriyolar Mikroenjeksiyon

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.

DsRNA Enjeksiyon RNAi Girişim içine Drosophila Embriyolar

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.

Bir In vivo</emŞu Protein Kompleksler yalıtmak için> Çapraz yaklaşım Drosophila Embriyolar

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.

Sıtma Sivrisinekler Gelişimi, doğurganlık ve Kalma bir Epigenetik İlaç Test Etkileri Toksikolojik Tahliller

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.

Sivrisinekler için roman böcek kimyaları toksisite testleri için iletişim kuralları

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.

Salya vektör yetkinlik sivrisinek analiz etmek için bir yöntem olarak zorla

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.

A Blood-Free Diyet Arka Anopheline Sivrisinekler için

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