Data from: Quantification of sporozoite expelling by Anopheles mosquitoes infected with laboratory and naturally circulating P. falciparum gametocytes
Data files
Feb 28, 2024 version files 123.88 KB
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Data_for_Figure_1B_oocyst_density_and_prevalence.xlsx
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Data_for_Figure_2_infection_intensity_and_time_to_sg_positivity.xlsx
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Data_for_Figure_2B_Sporozoite_densities_increase_with_oocyst_age.xlsx
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Data_for_Figure_3_Artificial_skin_feeding_cultured_gametocytes.xlsx
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Data_for_Figure_4_Artificial_skin_feeding_natural_infected_Burkina_Faso.xlsx
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Data_for_Figure_5_Clonal_complexity_of_P._falciparum_infections_in_Burkina_Faso.xlsx
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README.md
Abstract
It is currently unknown whether all Plasmodium falciparum infected mosquitoes are equally infectious. We assessed sporogonic development using cultured gametocytes in the Netherlands and naturally circulating strains in Burkina Faso. We quantified the number of sporozoites expelled into artificial skin in relation to intact oocysts, ruptured oocysts, and residual salivary gland sporozoites. Sporozoites were quantified by highly sensitive qPCR; intact and ruptured oocysts by fluorescence microscopy following antibody staining of circumsporozoite protein. In laboratory conditions, higher total sporozoite burden in mosquitoes was associated with a shorter duration of sporogony (p<0.001). Overall, 53% (116/216) of P. falciparum infected An. stephensi mosquitoes expelled sporozoites into artificial skin. The medians of expelled and residual salivary gland sporozoites were 136 (IQR: 34-501) and 23,947 (IQR: 9127-78,380), respectively. There was a strong positive correlation between ruptured oocyst number and salivary gland sporozoite load (ρ=0.8; p<0.0001) and a weaker positive correlation between salivary gland sporozoite load and the number of sporozoites expelled (ρ=0.35; p=0.0002). In Burkina Faso, An. coluzzii mosquitoes were infected by natural gametocyte carriers. Among mosquitoes that were salivary gland sporozoite positive, 89% (33/37) expelled sporozoites with a median of 1035 expelled sporozoites (IQR: 171-2969) and harbored a median of 45,100 residual salivary gland sporozoites (IQR: 20,310-164,900). Again, we observed a strong correlation between ruptured oocyst number and salivary gland sporozoite load (ρ=0.9; p<0.0001) and a positive correlation between salivary gland sporozoite load and the number of sporozoites expelled (ρ=0.7; p<0.0001). Mosquito salivary glands in Burkina Faso harbored 1-3 distinct parasite clones; several mosquitoes expelled multiple parasite clones during probing.Whilst sporozoite expelling was regularly observed from mosquitoes with low infection burdens, our findings indicate that mosquito infection burden is associated with the number of expelled sporozoites. Future work is required to determine the direct implications of these findings for transmission potential.
README: "Quantification of sporozoite expelling by Anopheles mosquitoes infected with laboratory and naturally circulating P. falciparum gametocytes "
https://doi.org/10.5061/dryad.dbrv15f89
Datasets are linked to individual display figures in the article.
Description of files
File Name: Data_for_Figure_1B_oocyst_density_and_prevalence.xlsx
Description of file:
Data that is generated in mosquito feeding assays with cultured gametocytes in Nijmegen The Netherlands. The set contains three conditions: standard, routine feedings for generating infectious mosquitoes; EIP, feedings performed to conduct EIP experiments in the paper aiming for with low and high infected mosquitoes; Skin, feedings performed to conduct Skin experiments in the paper aiming for low and high infected mosquitoes.
Variables:
- Condition: Standard, Skin, EIP
- Oocyst density: Mean of oocysts counted in at least 10 mosquitoes that derived from the same cage
- Prevalence infection %: Mean percentage of infected mosquitoes that contained at least one oocyst
File Name: Data_for_Figure_2_infection_intensity_and_time_to_sg_positivity.xlsx
Description of file:
Generated data for low and high infected mosquitoes, with cultured gametocytes, processed on different days post infection to determine EIP for mosquitoes reared under laboratory/controlled conditions (Nijmegen, The Netherlands).
Variables:
- Day: Day post infection, either day 9, 10 or 11
- Experiment: Data collected in independent experiments number 6, 7 or 8
- SG load: Load/number of SG (Salivary gland) SPZ by targetting COX-1 in qPCR
- SG (pos=1/neg=0): Positive or negative salivary gland
- Mosquito Carcass load: Signal/load - number of parasites - measured in the carcass
- Total parasite load: Signal/load - number of parasites - measured in dissected salivary glands
File Name: Data_for_Figure_2B_Sporozoite_densities_increase_with_oocyst_age.xlsx
Description of file:
Data collected on day 9 or day 10 post infection, with mosquitoes infected with cultured gametocytes in Nijmegen The Netherlands. Infected mosquitoes with only one oocyst were selected for this experiment. The oocyst was removed from midgut and processed in qPCR targetting COX1.\
Variables:
- Day: Day post infection, either day 9 or 10
- Single oocyst: midgut containing 1 oocyst
- Sporozoites density: Density of sporozoites within one oocyst, quantified by qPCR targetting COX1
File Name: Data_for_Figure_3_Artificial_skin_feeding_cultured_gametocytes.xlsx
Description of file:
Data generated for low and high infected mosquitoes with cultured gametocytes (strain NF54 and NF135), in Nijmegen The Netherlands.
Variables:
- Sample Number: Labeling of collected samples
- Skin Exp: Experiment, group of infected mosquitoes within the same experiment all processed on the same day
- Strain: NF54 or NF135
- Condition (feed): Describes if the mosquito ingested a blood meal and if it was fully fed (FF), partially fed (PF), unfed (UN) or only probed (unfed but probing)
- Oocyst sheet: Number of oocysts sheets observed after immunolabeling
- Intact Oocyst: Number of intact oocysts observed after immunolabeling
- SG load (COX1): Dissected salivary glands processed by qPCR targetting COX1 for quantification of SPZ density
- Skin (COX1): Processed skin and quantified SPZs by qPCR targetting COX1
- Notes: Description of labeling
File Name: Data_for_Figure_4_Artificial_skin_feeding_natural_infected_Burkina_Faso.xlsx
Description of file:
Data generated with infected mosquitoes, using natural infected gametocytes carriers in Burkian Faso. The skin feeding data is presented on tab one. To generate high infected mosquitoes, wth low density cariers, gametocytes were enriched with MACS, donor/MACS information can be found on tab 2.
tab: Skin Feedings
tab: all MACS experiments
Variables:
tab: Skin Feedings
- Sample No. : Labeling of collected samples
- Skin Exp: Experiment, group of infected mosquitoes within the same experiment all processed on the same day
- Strain: Donor used to infected mosquitoes
- Condition (feed): Describes if the mosquito ingested a blood meal and was fully fed (FF), partially fed (PF), unfed (UN) or probed (unfed but probing)
- Oocysts sheets: Number of oocysts sheets observed after immunolabeling
- Oocyst intact: Number of intact oocysts observed after immunolabeling
- Oocyst(sheets+intact) combined : Total number of oocysts present on the midgut
- sg_load: Dissected salivary glands processed by qPCR targetting COX1 for quantification of SPZ density
- sporo expelled into skin: Processed skin for quantification of expelled SPZs, by qPCR targetting COX1
- total sporo load skin + SG: Total number of spz of salivary gland and skin sample combined
- notes: Description of labeling
File Name: Data_for_Figure_5_Clonal_complexity_of_P._falciparum_infections_in_Burkina_Faso.xlsx
Description of file:
Data generared for identifying clonality of infection from natural gametocyte cariers in Burkina Faso, that were used to infected mosquitoes for conducting skin feeding experiments.\
Variables:
- ID: Sample numbering according to centre in Burkina Faso
- Donor: Donor used to infected mosquitoes
- where: Sample from mosquito salivary gland (SG) or Skin
- density: SG spz density of parasites quantified by qPCR targetting COX1
- CloneA: Clone with unique DNA sequence, labeled A
- CloneB: Clone with unique DNA sequence, labeled B
- CloneC: Clone with unique DNA sequence, labeled C
- CloneD: Clone with unique DNA sequence, labeled D
- CloneE: Clone with unique DNA sequence, labeled E
- CloneF: Clone with unique DNA sequence, labeled F
- CloneG: Clone with unique DNA sequence, labeled G
- CloneH: Clone with unique DNA sequence, labeled H
- CloneI: Clone with unique DNA sequence, labeled I
- CloneJ: Clone with unique DNA sequence, labeled J
- Notes:Description of labeling
Methods
This work involves data collected with in vitro cultured gametocytes and gametocytes circulating in naturally infected individuals. Parasite development in mosquitoes was examined in relation to parasite burden in mosquitoes. Data collected centered around the expelling of sporozoites by mosquitoes infected with high and low infection burden. Parasites were quantified primarily by qPCR, supplemented by microscopy.