Methods in Anopheles Research

Second Edition (2011)

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Preface

Preface to the Second Edition  

Table of Contents

Chapter 1 : Insectary Operation

1.1 Equipping and Operating an Insectary

1.2 Cleanliness and General Maintenance

1.3 Scheduling and Regulating Your Work Load

1.4 Maintaining Stock Purity

1.5 Insectary Manager Responsibilities

Chapter 2 : Anopheles Laboratory Biology and Culture

2.1 Behavior and Physiology of Anophelines in the Laboratory

2.2 Infections in Mosquito Cultures

2.3 Modifying Fecundity, Longevity and Size

2.4 Anopheles Culture

2.4.1 Collecting Anopheles Eggs

2.4.2 Bleaching Anopheles Eggs

2.4.3 Hatching Anopheles Eggs

2.4.4 Determining Egg Hatch Rates

2.4.5 Estimating the Number of Eggs and Larvae

2.4.6 Anopheles Larval Culture

2.4.7 Separating Larvae and Pupae

2.4.8 Anopheles Adult Caging

2.4.9 Anopheles Adult Diet

2.4.10 Bloodfeeding : Membrane Apparatuses and Animals

2.5 Basic Anopheles Mendelian Genetics

2.6 Basic Anopheles Population Genetics

Chapter 3 : Specific Anopheles Techniques

3.1 Determining the Sex of Anopheles Pupae and Larvae

3.2 Microinjection Methods for Anopheles Embryos

3.3 Plasmodium Sporozoite ELISA

3.4 Family Culture

3.5 Determining Egg Hatch Rates

3.6 Anopheles Mating

3.6.1 Mating : General Considerations

3.6.2 Forced Copulation

3.6.3 Pair Matings

3.6.4 An. gambiae and arabiensis Mating Status Determination

3.7 Anopheles Embryo Fixation

3.8 Mosquito Anesthesia

3.9 Eye Color Mutant Screening

3.10 Establishing Cell Lines from Anopheles spp. Embryonic Tissues

3.11 Molecular Identification of Plasmodium spp. in Anophelines
3.12      96 Well Genomic DNA Extraction
3.13      Determination of Lipid, Glycogen and Sugars in Mosquitoes
3.14      Vector Population Monitoring Tool using Real Time PCR
3.14.1   An. gambiae s.l. species complex ID assay
3.14.2   Plasmodium detection assay
3.14.3   Knock down resistance (kdr) assays
3.14.4   Insensitive acetylcholinesterase (iAChE) assay
3.14.5   Resistance to dieldrin (rdl) assay
3.15      Molecular ID of Mammalian Blood Meals
 

Chapter 4 : Stock Authentication

4.1 Authentication by Morphological Characteristics

4.2 Authentication by PCR

4.2.1 Anopheles gambiae complex (Scott et al.)

4.2.2 Anopheles gambiae complex (Wilkins et al.)

4.2.3 Anopheles funestus complex

4.2.4 Anopheles minimus complex

4.2.5 Anopheles quadrimaculatus complex (Cornel et al.)

4.2.6 Anopheles quadrimaculatus complex (Levine et al.)

4.2.7 Anopheles gambiae Tep 1 Assay

4.2.8 Anopheles ITS2 Amplification

4.2.9 An. gambiae Ribosomal DNA Type (Fanello et al.)

4.2.10 An. gambiae Ribosomal DNA Type (Wilkins et al.)

4.2.11 Anopheles arabiensis ND5 Molecular Authentication

4.2.12 Anopheles gambiae 2La inversion (White et al.)

4.2.13 Anopheles gambiae White gene

4.2.14 Combined Anopheles gambiae ID and rDNA type Assay

4.3 Authentication by Bioassay

4.3.1 Larval Insecticide Resistance Assays

4.3.2 WHO Paper Testing

4.3.3 CDC Bottle Bioassays

4.3.4 Microplate Enzyme Activity Assays Introduction

4.3.5 Microplate Insensitive Acetylcholinesterase Assay

4.3.6 Microplate Glutathione s-transferase Assay

4.3.7 Microplate Nonspecific Esterase Assay

4.3.8 Microplate Oxidase Assay

4.4 Insecticide Resistance Allele Assay by PCR

4.4.1 Knockdown Resistance - Anopheles gambiae

4.4.2 Dieldrin Resistance - An. gambiae and An. arabiensis

4.4.3 Knockdown resistance in Anopheles gambiae - Huynh et al.

4.4.4 ACE-1 resistance in An. gambiae

Chapter 5 : Dissection Techniques

5.1 General Dissection Buffers

5.2 Rapid Larval Midgut Extraction

5.3 Adult Male Testes Dissection

5.4 Dissecting Plasmodium-infected Mosquitoes

5.4.1 Midgut

5.4.2 Salivary Glands

5.5 Examination of Ovaries by Tracheal Distension to Determine Parity

5.6 A. gambiae s.l. Ovarian Polytene Chromosome Preparation

5.7 A. gambiae s.l. Salivary Gland Chromosome Preparation

5.8 Dissecting Spermathecae to Determine Insemination Status

5.9 Larval Midgut Vivisection

Chapter 6 : Obtaining Live Material

6.1 Permits and Regulations