Dear Devinder,

I guess with the MR4 protocol, from page 117-121, you will have enough information to start working with your Anopheles colony. Now regarding A. aegypti, here are my observations.

When rearing the larvae, always keep them in an uncrowded condition with enough food available. By doing this, you will for sure get larger females which will blood feed more and lay more eggs, also, your males will also be bigger, which will then mate with a higher success rate.

Here's a scheme that you can test and see if it works for you.

Wednesday: put your eggs to hatch in dechlorinated water + a little bit of food (keep in mind that Aedes larvae do not feed properly in the water surface like Anopheles, so use proper food source).

Thursday: separate your larvae in more than one box to avoid crowded conditions.

Friday: add food

Saturday+Sunday: add food (keep in mind to avoid adding too much food, otherwise your larvae will die! Also, always change the water when it gets too dirt)

next week, from Monday to Friday: collect pupae and prepare your cages (keep in mind that usually the first pupae are always male, so when dividing the cages, try to mix these initial pupae into different cage; or you can separate them by sex, using a Stereo Microscopes if you want something more accurate.

by doing this, you should have collected all the pupae until Thursday. Now when it comes to blood feeding these mosquitoes.

You probably know that males take around 3 days to start copulating, so you cannot blood feed the females before that period of time and expect a good batch of viable eggs. So, regarding our little scheme, here's what you should do.

Leave the mosquitoes copulating with sugar solution available until monday.

Monday: you will take out the sugar and leave the females without it for 24 hours.

Tuesday: now you will blood feed the cages. (Here we use human blood). After this process, put again sugar solution in the cage.

Now do nothing until Friday, which would be a good time for the females to "maturate their eggs".

Friday: put inside the cages your cups with water in it and a strip of paper for the females to lay their eggs and leave them there until monday which is the day you can take them out and let them dry (you must let them dry!) until Wednesday when you can restart the process.

This Scheme that I just told you about, is regarding a discrete format of colony maintenance, not a colony with overlapping generations!

Sorry about any English mistake, hope it helps!

Heverton

which mosquito will it be? Because there's major diferences when rearing for example Aedes aegypti, Anopheles aquasalis, Aedes fluviatilis and so on...

Knowing your desired mosquito, I can give you some advices if you want to, but in advance, if you are going to work with Anopheles for example, this will help you get start:

http://www.mr4.org/Portals/3/Methods_In_Malaria_Research-5theditionv5-2.pdf

Thank you Mr Heverton Dutra for your answer and providing the related site.

Actually i have just started this work under a project dealing with biological control aspects of mosquitoes including Anopheles, Aedes and Culex. If you can advice me for mosquito rearing, especially we are having problem in egg laying part.

Dear Devinder,

I guess with the MR4 protocol, from page 117-121, you will have enough information to start working with your Anopheles colony. Now regarding A. aegypti, here are my observations.

When rearing the larvae, always keep them in an uncrowded condition with enough food available. By doing this, you will for sure get larger females which will blood feed more and lay more eggs, also, your males will also be bigger, which will then mate with a higher success rate.

Here's a scheme that you can test and see if it works for you.

Wednesday: put your eggs to hatch in dechlorinated water + a little bit of food (keep in mind that Aedes larvae do not feed properly in the water surface like Anopheles, so use proper food source).

Thursday: separate your larvae in more than one box to avoid crowded conditions.

Friday: add food

Saturday+Sunday: add food (keep in mind to avoid adding too much food, otherwise your larvae will die! Also, always change the water when it gets too dirt)

next week, from Monday to Friday: collect pupae and prepare your cages (keep in mind that usually the first pupae are always male, so when dividing the cages, try to mix these initial pupae into different cage; or you can separate them by sex, using a Stereo Microscopes if you want something more accurate.

by doing this, you should have collected all the pupae until Thursday. Now when it comes to blood feeding these mosquitoes.

You probably know that males take around 3 days to start copulating, so you cannot blood feed the females before that period of time and expect a good batch of viable eggs. So, regarding our little scheme, here's what you should do.

Leave the mosquitoes copulating with sugar solution available until monday.

Monday: you will take out the sugar and leave the females without it for 24 hours.

Tuesday: now you will blood feed the cages. (Here we use human blood). After this process, put again sugar solution in the cage.

Now do nothing until Friday, which would be a good time for the females to "maturate their eggs".

Friday: put inside the cages your cups with water in it and a strip of paper for the females to lay their eggs and leave them there until monday which is the day you can take them out and let them dry (you must let them dry!) until Wednesday when you can restart the process.

This Scheme that I just told you about, is regarding a discrete format of colony maintenance, not a colony with overlapping generations!

Sorry about any English mistake, hope it helps!

Heverton

Thank you Dear Dutra

I will follow this scheme and share it with you in future. Let's hope for its success.

Good day

Hi Devinder! I think Sir Heverton's scheme is very useful. I work with Aedes aegypti and albopictus in the lab and here's some tips I gained from experience.

We use brewer's yeast as feed (1/4 of a tablet for early instars and 1/2 to 1 tablet for 3rd to 4th instars daily depending on the number of larvae per container), some add liver powder for this.

Next, Aedes larval set-ups, unlike Culex', are very prone to clouding due primarily to overfeeding. so it is necessary to change the water when it starts to cloud so always keep a supply of dechlorinated tap water.

For bloodfeeding, I use a live guinea pig. I Shave the back part so that the mosquitoes can readily bite. Some use human blood which you can get from blood banks (those cannot be used for transfusion) or pig or cow blood placed on a hemotek system or a pig intestine/ condom/ sausage wrap. It is necessary to heat the blood to induce feeding. I use the guinea pig since it is more convenient for me since I am maintaining a smaller colony. I keep a black cloth over the cages too.

For oviposition, I line the container containing the pupae inside the rearing cage with filter paper where the females can lay. You can also place a separate container though. Aedes eggs should be dried, but do not dry it right after taking the filter papers out. Leave it moist for about 24 hours just to finish embyronic development. When drying, do not desiccate too much and store it in an airtight (but not too dry) container.

My egg collections are usually viable for one to two months after storage. Viability decreases after.

I hope this helps! Best of luck

Thank you dear Justine, i will try this. I am using rat for blood feeding.

Dear Devinder,

Her is our routine work in very simplified version of MR4 rearing guideline it may help you.

Regards