إعداد مقدما من microinjection : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> البعوض الدم : للحقن من الاثنين إلى الأربعاء الإناث إطعام في يوم الجمعة السابق. لحقن الخميس والجمعة ، تتغذى على الإناث يوم الاثنين من الأسبوع نفسه. </li><li style=";text-align:right;direction:rtl"> تعد الإبر كوارتز باستخدام برنامج (2) والعازلة متساوي التوتر (انظر المواد). </li><li style=";text-align:right;direction:rtl"> &quot;مد أنبوب&quot; للأجنة ذبابة الفاكهة هي ثقافة العادية القارورة. الرطب القطن والصوف في الجزء السفلي ، مع قرص من الورق الرطب تصفية تغطية هذه الأزمة. </li><li style=";text-align:right;direction:rtl"> إعداد زلة تغطية البلاستيك عن طريق الالتزام المزدوج الجانب الشريط إلى نهاية واحدة. الشريط تقليم لتغطية زلة بحيث تنتهي عند حافة الانزلاق الغطاء. </li><li style=";text-align:right;direction:rtl"> يعد النفط للجفاف. </li><li style=";text-align:right;direction:rtl"> إعداد طبق بيتري مع متساوي التوتر عازلة لنقل الأجنة لالفقس. </li></ol> إعداد زرع القسري : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> جمع 60-10 الإناث تغذية الدم مع استخدام لالشافطة ونقلها إلى القارورة الثقافة ذبابة الفاكهة مع القطن ورقة الترشيح الرطب مع العازلة متساوي التوتر. </li><li style=";text-align:right;direction:rtl"> ثم يتم وضع مرة أخرى في ظروف البعوض insectery في الظلام ، وسمح لوضع البيض لمدة 1 ساعة و15 دقيقة. </li><li style=";text-align:right;direction:rtl"> اسمحوا البالغين تطير في القفص ، وإزالة القرص رقة الترشيح مع الأجنة. </li><li style=";text-align:right;direction:rtl"> ليصطف والبيض ، وذلك ضمن نطاق تشريح. جمع باقات من البيض مع الفرشاة الجميلة (سابل ، لا 0000) ونقل إلى مربع أعد ورقة Whatman 3MM غارقة مع العازلة متساوي التوتر. ابقاء ورقة مبللة كل مرة. لا تدع الحصول على البيض المجفف. إزالة مشيماء أديمية ظاهرة مع الفرشاة ، فإنه يساعد على عصا أفضل البيض إلى الشريط. </li><li style=";text-align:right;direction:rtl"> باستخدام غرامة forcep رقم 5 أو الفرشاة الجميلة ، والتقاط الأجنة الرمادية الداكنة وترتيبها في خط على مربع من Whatman 3MM الرطب مع العازلة متساوي التوتر. يصطف 20-30 الأجنة. يجب أن تكون جميع الأجنة في نفس اتجاه الحقن يجب أن يكون في القطب الخلفي. نهاية الأمامي من الأجنة هو أوسع قليلا من الخلفية. ثم ، باستخدام شرائط من الورق Whatman 3MM جافة والتصفية حيث تصطف البيض بنسبة يضغط بشدة على كل من جانبي خط البيض ، وعدم لمس البيض. </li><li style=";text-align:right;direction:rtl"> لنقل البيض ، وعكس الشريحة التي تحتوي على شريط مزدوج الوجهين (الطب) ، واضغط بلطف ضد البيض. نهاية الخلفي من البيض يجب أن تكون قريبة جدا من حافة الشريط جانب ومزدوجة. والبلل ورقة هو أمر حاسم لهذا ، إذا كانت رطبة جدا انهم لن عصا. أفعل جفاف الأجنة من المركز الثاني 50-10 ، ولكن الوقت من الجفاف يعتمد على الرطوبة ودرجة الحرارة في الغرفة microinjection. </li><li style=";text-align:right;direction:rtl"> أمر حاسم لتجفيف البيض : قليلا الكثير من الأجنة وسوف لا يفقس. الحمض النووي من دون تجفيف ليس الخوض في الجنين. </li><li style=";text-align:right;direction:rtl"> تغطي الأجنة المجفف مع النفط لمنع جفاف الهالوكربون أخرى. </li></ol> Microinjection الأجنة : وأهم جانب من جوانب جيدة الحقن هي نوعية الإبرة (انظر الملاحظة). <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> شغل الإبرة مع الحل القائم على الحمض النووي ليتم حقنها باستخدام microloader (956،003 إيبندورف # 5242). وهناك حاجة soliution جدا حقن قليلا ، من 1 إلى 2 مل. </li><li style=";text-align:right;direction:rtl"> الاتصال الإبرة إلى Transjector إيبندورف ، التي تسيطر على الحقن وضغط الوقت ، وكذلك في الضغط الخلفي. يتم تنفيذ Microinjection باستخدام المجهر مع مرحلة الانتقال (لايكا) في التكبير 10 x و micromanipulator (لايكا). إذا لزم الأمر ، قد تكون على استعداد مرحلة مجهر التي أثارها التراص الشرائح المجهر 4-5. ضع الغطاء زلة تحمل أجنة على المرحلة التي أثيرت. يتم حقن الاجنة بزاوية · 150 ؛ اختراق ينبغي أن يكون في القطب الخلفي. عن طريق الحقن ، وأظل الإبرة ثابتة وتحريك مرحلة من مراحل المجهر. عندما الإبرة هو جديد ، هو غيض مختومة وفواصل عادة في الحقن الأولى. أنا أعمل دائما من الضغط اللازم لطرد قطرات صغيرة من الحمض النووي في النفط في الفترات الفاصلة بين كل حقنة. </li><li style=";text-align:right;direction:rtl"> أنا وضعت وقت الحقن ل0،2-0،9 ثانية وهكتوبسكال 1000 عندما إبرة جديدة. أنا يختلف الضغط والوقت حتى يتم النظر قطرات صغيرة تخرج من الإبرة إلى النفط. من الضغط الخلفي يحتاج إلى تعيين نحو 100. كما يحصل البالية طرف الإبرة ، وضغط الحقن لابد من تخفيض للحفاظ على انخفاض حجم الحقن (حوالي 300 هكتوبسكال). بعد هذا ، غيض إبرة كبيرة جدا وتتسبب في مقتل أكثر من الأجنة ل. </li><li style=";text-align:right;direction:rtl"> بعد الحقن ، وإزالة النفط مع المناديل الورقية وتغطية الشريحة مع الأجنة وضعها في طبق بتري متساوي التوتر مع العازلة. ضع الأطباق في insectory وتركهم هناك حتى الأجنة يفقس. أنها بداية ليفقس بعد 2-6 أيام. نقل اليرقات في الماء المقطر مع قليل من الغذاء السمكي الأرض. </li></ol> تلاحظ تم عزل الحمض النووي البلازميد حل للحقن باستخدام طقم البلازميد EndoFree : الحمض النووي للحقن.عجلت بناء البلازميد ومزيج مساعد البلازميد معا في تركيز الحق ، مع الأيزوبروبانول. وكان بيليه تغسل مع الايثانول 70 ٪ قبل إعادة التعليق في المخزن الحقن. قبل الحقن ، باستخدام الحمض النووي نظيفة Millex GV - العمود. Izotonic العازلة : <table border="1" cellpadding="8" style=";text-align:right;direction:rtl"><tbody><tr><td> كلوريد الصوديوم 1.68 M -- 88.3 مل كلوريد الصوديوم 1.68 م -- 2.9 م 1 مل M Hepes -10.7 1.12 M CaCl 2-2،2 مل DH 2 O -- 896 مل </td><td> OR </td><td> 5 M كلوريد الصوديوم -- 29.67 مل 1 م بوكل -- 4.87 مل 1 Hepes M -- 10.7 مل 1.12 M CaCl 2-2،2 مل DH 2 O -- 952.56 مل </td></tr></tbody></table> التكيف مع درجة الحموضة 7.2

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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

إدخال جينات دخيلة في الجينوم من البعوض microinjection يتطلب تقنيات مصممة خصيصا لأنواع محددة من الفائدة. هذا البروتوكول الفيديو يوضح الطريقة المستخدمة من قبل المختبر جيمس microinject يبني الحمض النووي في الأجنة stephensi الأنوفيلة لجيل من البعوض المتحول. وتتجلى أساليب إعداد microinjection الإبر ، وجمع واعداد وتنفيذ الأجنة microinjection.

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

الأنوفيلة البعوض stephensi نواقل الملاريا في الهند والتي تسكن في جميع أنحاء آسيا. هذا الفيديو يوضح هذه التقنية لأداء microinjections من هذا النوع مع الجينات المحورة التي من شأنها أن تضفي على الملاريا المقاومة للبعوض. الكثير من منهجية تظاهروا في هذا الفيديو هو تطبيق لتقنيات microinjection أنواع أخرى من البعوض.

حقن آن. stephensi الجنين لتوليد الملاريا مقاومة البعوض

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 Views.  University of California, Irvine (UCI). الأنوفيلة البعوض stephensi نواقل الملاريا في الهند والتي تسكن في جميع أنحاء آسيا. هذا الفيديو يوضح هذه التقنية لأداء microinjections من هذا النوع مع الجينات المحورة التي من شأنها أن تضفي على الملاريا المقاومة للبعوض. الكثير من منهجية تظاهروا في هذا الفيديو هو تطبيق لتقنيات...

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.

Microinjection الجنين بعوض ألف الحصول على البعوض المعدلة وراثيا

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

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.

في الرحم وحقن داخل البطيني Electroporation الجنين الجرذ 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. 

في الرحم وحقن داخل البطيني Electroporation من أجنة الفئران 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 الجنين اسماك الزرد إلى تحليل وظائف الجينات

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.

رني التدخل عن طريق الحقن في الرنا المزدوج الجديلة ذبابة الفاكهة الأجنة

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.

و في الجسم الحي يشابك النهج لعزل البروتين المجمعات من ذبابة الفاكهة الأجنة

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.

فحوصات السمية للاختبار تأثيرات جينية من المخدرات للتنمية، الخصوبة والوراثة من الملاريا البعوض

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.

بروتوكولات لاختبار السمية لرواية كيمياء مبيدات الحشرات للبعوض

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.

أجبر اللعاب كأسلوب لتحليل اختصاص البعوض ناقل

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.

نظام غذائي خالي من الدم لخلفية البعوض الأنوفيلين

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