You can use a few different keys (Rioux et al. 1959, SCHAFFNER. -Clé de détermination des femelles dì\edes 1993) for morphologycal identification. I think that the method of storage is not suited to genetic analysis. Perhaps you should keep them low temperature.

Hi,

do you have any idea about the species present in the area? In my opinion the best approach should be to use ITS to design primers.

Anyway, maybe these paper could be a good starting point 

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4036727/

http://www.biotechniques.org/students/2014/marly/paper

Hoping it can help

Hi, most adult mosquitoes can be identified using the keys in Mosquitoes and their control by Becker et al http://www.amazon.com/Mosquitoes-Their-Control-Norbert-Becker/dp/3540928731

However, dry mosquitoes can be quite tricky to identify morphologically (they tend to fall apart when handling them). Why are you storing them in silica gel?

If you have trouble with identification we can help you at IRD in Montpellier.

You should use: Fascicule de  détermination des principales espèces de moustiques du littoral méditerranéen français, Sinegre, Rioux & Salgado

Hi Jessika,

I recommend you to extract DNA as soon as possible. After that, freezing at -80°C in small aliquots for storing. Then, thaw one tube and use it for further molecular assay. Maybe, you can find useful info for you on http://ecdc.europa.eu/en/publications/Publications/TER-Mosquito-surveillance-guidelines.pdf 

For COI barcodes you can search on the BOLD database: http://www.boldsystems.org/index.php/Public_BarcodeIndexNumber_Home 

As an initial approach, you can use the software for identification of european mosquitoes freely available on the MIVEGEC website:

http://www.mivegec.ird.fr/index.php?option=com_content&view=article&id=240&Itemid=237&lang=en

If you are in doubt, I would also suggest that you ask to entomologists from Montpellier (IRD).

Hi Jessika,

you can use any of above mentioned identification keys to identify species. Only for Anopheles (malaria) mosquitoes I recommend sequencing of ITS2 region. There are many sequences of that region in GenBank to compare with. 

Hi Jessika,

I work at IRD of Montpellier, i'm agree with Guillaume, the software proposed by IRD is a good tool. I used this software to identify mosquitoes from "camargue". Use is very simple.

Romain

Hello Jessica,

May not exist identification keys for your study area, I recommend you do a review of the species reported for your area and do a query has this website I put down, to offload work to support the identification of your mosquitoes.

I work with mosquitoes for 7 years and how hard it is to identify, if you need help send me an email to this email [email protected].

I recommend you look an expert in your region mosquitoes in Bold Systems 3 and blast are not all species, so using barcoding do not recommend it, also can not identify all species with pictures, so it is best to seek someone to help you identify them, Dr. Ralph Harbach you can find in Research Gate, is the best working mosquitoes. I hope this information helps.

take care

http://www.mosquitocatalog.org

You can ask Dr. Sylvie Manguin, IRP or Dr. Ralph Harbach, Nat. History Museum, London for advice on the best morphological keys for your area. 

Check this reference. It will be helpful

http://onlinelibrary.wiley.com/doi/10.1111/j.1755-0998.2010.02835.x/full

Hi Jessica,

Morphological identification and molecular tools are very expensives time cunsoming, and limited by dichotomical identification key availibility and molecular gene sequence target knowledge. In such condition, other innovative emerging methods could be used like mosquito identification by MALDI-TOF MS.

For additional information, see publication about Parola team on pubmed

Best

JMB

Hi Jessica,

I think that at IRD in Montpellier (ciao Nil) you can find all the entomological competences you need, especially for your study area. In any case, my colleagues and I can make our expertises available to you for both morphological and molecular analyzes. You can find attached the keys for the identification of Italian mosquitoes that could be useful for your purpose.

Marco

Dear Jessica,

you can use the following morphological key :Schaffner F., Angel G., Geoffroy B., Hervy J.P., Rhaiem A. & Brunhes J. 2001. The Mosquitoes of Europe/Les moustiques d’Europe. An Identification and Training Programme (CDRom), Montpellier, France: IRD Éditions & EID Méditerranée. But you will have also to take in account the invasive species that could have appeared in the area since 2001 probably only Ae. albopictus.

I think that Francis Schaffner could help you ont both aspects of your work (mrophological identification, barecoding) [email protected].

regards

Dear Jessica,

Hi, as I know there are at least 30 species of Plasmodium which are the pathogens of bird malaria and there are different mosquito species belong to the genera Aedes, Ochlerotatus, Culex, Culiseta, and Anopheles as vectors worldwide. You have received very good comments before. I suggest preserving and mounting your mosquito samples in a good way (dry adult specimens using micro pin). You need adult samples in a good condition for a correct morphological identification especially for the genera Aedes, Ochlerotatus, Culex. You may need to provide the microscope slides of some male genitalia for correct identification of some species (Cx. pipiens, Cx. torrentium etc). After correct morphological identification you may transfer them to tubes for PCR. The best specimens for molecular study are those which are preserved in alcohol 90% (of course for a longtime preservation). There are enough keys for identification of mosquitoes in France and Europe, however you may find our keys to Iranian mosquitoes (about 70 species) useful in my profile at ResearchGate (Azari-Hamidian & Harbach 2009). The species Orthopodomyia pulcripalpis with some other species especially in the tribe Aedini which occur in France, are not in Iran, also some Iranian species do not exist in France, however I think the style and terminology in my keys will help you. Also, there are some data about barcoding in our publications (Azari-Hamidian et al 2009, Azari-Hamidian et al. 2010). I hope you find these data useful.

Thank you everyone for all your comments, they have been very helpful. I just had one more comment, I have read that alcohol can dissolve the DNA (especially malaria parasites) in the samples, especially if storing them for any extended period of time (I may be storing samples for up to 3 months before analysing them) reducing the possibility of carrying out effective PCR analysis, is there a general consensus that alcohol is the preferred storage technique? I was planning on storing them over silica gel desiccant as I have read this is better for preserving the DNA?

Many thanks

Dear Jessica,

As I know alcohol is a good preservative medium of the DNA of mosquitoes (in comparison to dry samples) for a longtime storage based on my experience at the Natural History Museum, London, however I do not know about DNA of Plasmodium.

Best

Hi Jessika, as recommended by WRBU ¨For all stages, the best method for preservation for DNA and protein analysis is in liquid nitrogen. For protein analysis it must be in liquid nitrogen. For preservation of DNA, 95-100% ethyl alcohol can be used very effectively without immediate need for refrigeration. Specimens are placed live into the alcohol followed by 1 or two changes of fresh alcohol. Specimens in alcohol should be placed at at least -70o when refrigeration is available. In alcohol the DNA remains in relatively good condition at room temperature for 1 or 2 months¨ http://www.wrbu.org/techniques.html 

Dear Jessica,

you can conserve your specimens over silica gel desiccant without problem with DNA if you entend to use molecular methods to asses the infection rate of the mosquitoes. If you want to detect plasmodium protein, you can use the same storage on the field but  you will have to freeze quickly your vials at -20°c. 

 Hi Jessica

Indeed the CD-Rom published at IRD is a good tool.

You can use it online (in French) at http://www.mivegec.ird.fr/index.php?option=com_content&view=article&id=240&Itemid=237&lang=en

There is also a dichotomic key in Becker, N., Petrić, D., Zgomba, M., Boase, C., Madon, M., Dahl, C. & Kaiser A. 2010. Mosquitoes and their control. Springer, Heidelberg Dordrecht London New York. 579 pp.

I can suggest you to use MALDi-TOF to confirm some morphological ID by sending them to Mabritec, Riehen, CH, where we have a large data base for well identified mosquito species (also within complex species). And if you wish you can send me some specimens for ID confirmation (I worked in Southern France during almost 10 years).

Good luck

Francis

Hello, we must preserve the specimens at -20 for both DNA to identify them, if preserved in alcohol, you can destroy DNA and mosquitoes lose setae and scales, useful for identification.

Dear Jessica,

The CD-Rom edited by IRD is indeed a valid tool (the invasive Aedes albopictus is included). You can use it online at http://www.mivegec.ird.fr/index.php?option=com_content&view=article&id=240&Itemid=237&lang=en

You could also use a dichotomic key published in Becker, N., Petrić, D., Zgomba, M., Boase, C., Madon, M., Dahl, C. & Kaiser A. 2010. Mosquitoes and their control. Springer, Heidelberg Dordrecht London New York

If you need confirmation of some specimens, you can (1) send some to myself for morphological ID, (2) submit to Maldi-TOF MS by sending to Mabritec, Riehen. CH, were we have a large data base for European species including for species complexes (in press) and which is cheaper than PCR+sequencing.

Good luck, and enjoy the field!

Francis

Check www.mosquitocatalog.org. Many resources are available there.

Hi to all

The interactibve Schaffner et al CR rom key is no longer available.

The link above is broken, The good one is (cf.) this:

http://www.mivegec.ird.fr/en/identiciels-en-gb-8

The keys I used online were:

http://bioinfo-web.mpl.ird.fr/identiciels/culeuropegenre/java/culeuropegenre.html

(european genera)

http://bioinfo-web.mpl.ird.fr/identiciels/culeurope/java/culeurope.html

(european species)

When I began to use them a couple of years ago, I have to add a few exceptions on java, as I posted here:

http://www.diptera.info/forum/viewthread.php?thread_id=72570

But with the more recent updates it seems impossible....

My informatics knowledge is quite limited, so any success will that matter will be wellcommed

Another possibility is the authors to edit the CD again,....

Best wishes,

Rafael