Description
Material provided: Whole mosquitoes, dissected tissues, or extracts
Unit definition: 50 infected females (25 if dissected) supplied as 100 fully-engorged females with infection prevalence at least 50%
Vector information
Family: Culicidae
Genus: Anopheles
Species: Anopheles coluzzii
Strain Name: N‘Gousso
Place of origin: Yaoundé, Cameroon
Date of colonisation: 2006
Genotype features: TEP1*S1
Phenotype features: susceptible to P. falciparum
Parasite information
Phylum: Apicomplexa
Genus: Plasmodium
Species: Plasmodium falciparum
Strain: Plasmodium falciparum isolate NF54 was originally isolated from an individual living near Schiphol airport, Amsterdam, The Netherlands.
Infectivity: Infection at 0.3% final gametocytaemia gives generally 40,000-80,000 spz / mosquito 14 days post infection.
Mycoplasmic content: Asexual cultures are regularly checked for Mycoplasma contamination.
Production protocol
Specific dsRNA will be provided by the end-user. A. coluzzii Ngousso mosquitoes are microinjected with specific dsRNA at requested time before infection. Injected mosquitoes are fed through membrane feeders at 37°C for 15 min on the NF54 gametocyte cultures with a final gametocytaemia ranging between 0.15 to 0.30%. Unfed mosquitoes are removed after feeding. Infected mosquitoes are kept thereafter in the secured S3 laboratory at 26°C for 1-15 days before dissection.
Please specify in your request:
Time post-infection: Please specify one time point post-infection to harvest the samples. The most common options are ookinete infection (18-24 h post-infection), oocyst infection (2-13 d post-infection) and sporozoite infection (14-15 d post-infection), but other times are possible by request, contact us.
Whole or dissected: Instead of 50 whole mosquitoes, material can be supplied from dissected material harvested at the appropriate time point: 25 dissected midguts or 25 dissected salivary glands or 25 dissected ovaries, with the corresponding carcasses.
Form of material:
- Whole mosquitoes or dissected tissues, fixed in 70% ethanol, formalin, or other fixative: useful for DNA extraction from 70% ethanol, immunohistochemistry, not useful for RNA experiments or proteomics. Shipped as non-infectious material.
- Whole mosquitoes or dissected tissues, homogenised in a lysis buffer such as Trizol: useful for RNA, DNA and protein experiments. Shipped as non-infectious material.
- Whole mosquitoes or dissected tissues, frozen at -80°C or stored in RNALater: useful for RNA, DNA and protein experiments. Shipped as non-infectious material.
- Other formats or preparations may be possible by request, contact us.
Production conditions: Material is generated using default gametocyte dose and incubation temperature used by the supplying facility. Other conditions may be possible upon request.
Sample size and replicates: For larger sample sizes, request more units. Please specify if independent infections are needed as biological replicates (e.g., for RNAseq transcriptional profiling experiments).
Publications
- Harris, C., Lambrechts, L., Rousset, F., Abate, L., Nsango, S.E., Fontenille, D., Morlais, I., and Cohuet, A. (2010). Polymorphisms in Anopheles gambiae immune genes associated with natural resistance to Plasmodium falciparum. PLoS Pathog 6, e1001112.
- Ponnudurai, T., Leeuwenberg, A. D. and Meuwissen, J. H. (1981) Chloroquine sensitivity of isolates of Plasmodium falciparum adapted to in vitro culture. Tropical and Geographical Medicine 33, 50-54.
For more information, please contact us.