Streamlining sporozoite isolation from mosquitoes by leveraging the dynamics of migration to the salivary glands.

Anopheles stephensi Density dependence Extrinsic incubation period Oocysts Plasmodium berghei Salivary glands Sporozoites

Journal

Malaria journal
ISSN: 1475-2875
Titre abrégé: Malar J
Pays: England
ID NLM: 101139802

Informations de publication

Date de publication:
13 Sep 2022
Historique:
received: 01 04 2022
accepted: 12 08 2022
entrez: 13 9 2022
pubmed: 14 9 2022
medline: 16 9 2022
Statut: epublish

Résumé

Sporozoites isolated from the salivary glands of Plasmodium-infected mosquitoes are a prerequisite for several basic and pre-clinical applications. Although salivary glands are pooled to maximize sporozoite recovery, insufficient yields pose logistical and analytical hurdles; thus, predicting yields prior to isolation would be valuable. Preceding oocyst densities in the midgut is an obvious candidate. However, it is unclear whether current understanding of its relationship with sporozoite densities can be used to maximize yields, or whether it can capture the potential density-dependence in rates of sporozoite invasion of the salivary glands. This study presents a retrospective analysis of Anopheles stephensi mosquitoes infected with two strains of the rodent-specific Plasmodium berghei. Mean oocyst densities were estimated in the midguts earlier in the infection (11-15 days post-blood meal), with sporozoites pooled from the salivary glands later in the infection (17-29 days). Generalized linear mixed effects models were used to determine if (1) mean oocyst densities can predict sporozoite yields from pooled salivary glands, (2) whether these densities can capture differences in rates of sporozoite invasion of salivary glands, and (3), if the interaction between oocyst densities and time could be leveraged to boost overall yields. The non-linear effect of mean oocyst densities confirmed the role of density-dependent constraints in limiting yields beyond certain oocyst densities. Irrespective of oocyst densities however, the continued invasion of salivary glands by the sporozoites boosted recoveries over time (17-29 days post-blood meal) for either parasite strain. Sporozoite invasion of the salivary glands over time can be leveraged to maximize yields for P. berghei. In general, however, invasion of the salivary glands over time is a critical fitness determinant for all Plasmodium species (extrinsic incubation period, EIP). Thus, delaying sporozoite collection could, in principle, substantially reduce dissection effort for any parasite within the genus, with the results also alluding to the potential for changes in sporozoites densities over time to modify infectivity for the next host.

Sections du résumé

BACKGROUND BACKGROUND
Sporozoites isolated from the salivary glands of Plasmodium-infected mosquitoes are a prerequisite for several basic and pre-clinical applications. Although salivary glands are pooled to maximize sporozoite recovery, insufficient yields pose logistical and analytical hurdles; thus, predicting yields prior to isolation would be valuable. Preceding oocyst densities in the midgut is an obvious candidate. However, it is unclear whether current understanding of its relationship with sporozoite densities can be used to maximize yields, or whether it can capture the potential density-dependence in rates of sporozoite invasion of the salivary glands.
METHODS METHODS
This study presents a retrospective analysis of Anopheles stephensi mosquitoes infected with two strains of the rodent-specific Plasmodium berghei. Mean oocyst densities were estimated in the midguts earlier in the infection (11-15 days post-blood meal), with sporozoites pooled from the salivary glands later in the infection (17-29 days). Generalized linear mixed effects models were used to determine if (1) mean oocyst densities can predict sporozoite yields from pooled salivary glands, (2) whether these densities can capture differences in rates of sporozoite invasion of salivary glands, and (3), if the interaction between oocyst densities and time could be leveraged to boost overall yields.
RESULTS RESULTS
The non-linear effect of mean oocyst densities confirmed the role of density-dependent constraints in limiting yields beyond certain oocyst densities. Irrespective of oocyst densities however, the continued invasion of salivary glands by the sporozoites boosted recoveries over time (17-29 days post-blood meal) for either parasite strain.
CONCLUSIONS CONCLUSIONS
Sporozoite invasion of the salivary glands over time can be leveraged to maximize yields for P. berghei. In general, however, invasion of the salivary glands over time is a critical fitness determinant for all Plasmodium species (extrinsic incubation period, EIP). Thus, delaying sporozoite collection could, in principle, substantially reduce dissection effort for any parasite within the genus, with the results also alluding to the potential for changes in sporozoites densities over time to modify infectivity for the next host.

Identifiants

pubmed: 36100902
doi: 10.1186/s12936-022-04270-y
pii: 10.1186/s12936-022-04270-y
pmc: PMC9472382
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

264

Informations de copyright

© 2022. The Author(s).

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Auteurs

Ashutosh K Pathak (AK)

Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA. ash1@uga.edu.
Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA. ash1@uga.edu.
Center for the Ecology of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA. ash1@uga.edu.

Justine C Shiau (JC)

Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.
Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA.
Center for the Ecology of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.

Blandine Franke-Fayard (B)

Malaria Research Group, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.

Lisa M Shollenberger (LM)

Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.
Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA.
Department of Biological Sciences, Old Dominion University, Norfolk, VA, 23529, USA.

Donald A Harn (DA)

Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.
Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA.

Dennis E Kyle (DE)

Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.
Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA.
Department of Cellular Biology, University of Georgia, Athens, GA, 30602, USA.

Courtney C Murdock (CC)

Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.
Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA.
Center for the Ecology of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.
Odum School of Ecology, University of Georgia, Athens, GA, 30602, USA.
Department of Entomology, Comstock Hall, Cornell University, Ithaca, NY, 14853, USA.

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