Case Report: Plasmodium vivax Sporozoite Melanization in the Midgut and Salivary Gland of the Malaria Vector Anopheles darlingi

Najara Akira Costa dos Santos Plataforma de Produção e Infecção de Vetores da Malária- PIVEM, Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Rondônia, Brazil;
Instituto Nacional de Epidemiologia da Amazônia Ocidental (INCT-EpiAMO), Porto Velho, RO, Brazil;

Search for other papers by Najara Akira Costa dos Santos in
Current site
Google Scholar
PubMed
Close
,
Alessandra da Silva Bastos Plataforma de Produção e Infecção de Vetores da Malária- PIVEM, Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Rondônia, Brazil;

Search for other papers by Alessandra da Silva Bastos in
Current site
Google Scholar
PubMed
Close
,
Jéssica Evangelista Araújo Plataforma de Produção e Infecção de Vetores da Malária- PIVEM, Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Rondônia, Brazil;
Programa de Pós-graduação em Biologia Experimental – Universidade Federal de Rondônia/Fiocruz Rondônia, Porto Velho, Rondônia, Brazil;

Search for other papers by Jéssica Evangelista Araújo in
Current site
Google Scholar
PubMed
Close
,
José Daniel Costa Pontual Plataforma de Produção e Infecção de Vetores da Malária- PIVEM, Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Rondônia, Brazil;

Search for other papers by José Daniel Costa Pontual in
Current site
Google Scholar
PubMed
Close
,
Jansen Fernandes Medeiros Plataforma de Produção e Infecção de Vetores da Malária- PIVEM, Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Rondônia, Brazil;
Instituto Nacional de Epidemiologia da Amazônia Ocidental (INCT-EpiAMO), Porto Velho, RO, Brazil;

Search for other papers by Jansen Fernandes Medeiros in
Current site
Google Scholar
PubMed
Close
,
Joseph Michael Vinetz Section of Infectious Diseases, Department of Internal Medicine, Yale School of Medicine, New Haven, Connecticut, United States of America;

Search for other papers by Joseph Michael Vinetz in
Current site
Google Scholar
PubMed
Close
, and
Maisa da Silva Araujo Plataforma de Produção e Infecção de Vetores da Malária- PIVEM, Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Rondônia, Brazil;
Instituto Nacional de Epidemiologia da Amazônia Ocidental (INCT-EpiAMO), Porto Velho, RO, Brazil;
Programa de Pós-Graduação em Conservação e uso de Recursos Naturais – PPGReN, Fundação Universidade Federal de Rondônia, Porto Velho, Rondônia, Brazil;
Laboratório de Pesquisa Translacional e Clínica, Centro de Pesquisa em Medicina Tropical, Porto Velho, Rondônia, Brazil

Search for other papers by Maisa da Silva Araujo in
Current site
Google Scholar
PubMed
Close

ABSTRACT.

Anopheles darlingi is the primary malaria vector in the Amazon region and is highly susceptible to both Plasmodium vivax and Plasmodium falciparum parasites. Although anopheline mosquitoes may develop melanotic encapsulation in response to Plasmodium parasites, there is no record of An. darlingi exhibiting a melanization response to P. vivax, the main malaria parasite in the Americas. Here, we report the occurrence of P. vivax sporozoite melanization in An. darlingi mosquitoes.

INTRODUCTION

Malaria remains a significant infectious disease, responsible for approximately 250 million cases worldwide each year. In the Americas, Plasmodium vivax is the primary pathogenic, accounting for more than 70% of malaria cases,1 and Anopheles darlingi is the principal malaria vector in the Amazon region. Anopheline mosquitoes develop various immune responses against Plasmodium and other pathogens,2 including melanization, a complex humoral innate immunity response related to wound healing and pathogen encapsulation.3 Here, for the first time, we report cases of P. vivax sporozoite melanization in An. darlingi and investigate whether a higher intensity of infection improves melanization responses against sporozoites in An. darlingi.

CASES

Melanized sporozoites were observed in individuals of the An. darlingi species during the routine dissection of salivary glands (SGs) for sporozoite purification at 14 days post infection (dpi). Records of melanized sporozoites were found in bunches (Figure 1A–C) or individually (Figure 1E and F), both outside the SGs and outside the midgut (MG) of infected mosquitoes (Figure 1D). These mosquitoes were obtained from a Brazilian An. darlingi colony that has been maintained under laboratory rearing conditions since 20184 and were fed on blood samples from P. vivax–positive patient donors by direct membrane feeding assay (DMFA).

Figure 1.
Figure 1.

Melanized Plasmodium vivax sporozoite in Anopheles darlingi mosquitoes. (A and D) Melanized sporozoites outside the midgut. (B, C, E, and F) Melanized and nonmelanized sporozoites outside the salivary glands. White arrowheads indicate melanized sporozoites, and red arrowheads indicate free sporozoites. The white box indicates the total magnification under light microscopy. All images were obtained at 14 days post infection.

Citation: The American Journal of Tropical Medicine and Hygiene 110, 3; 10.4269/ajtmh.23-0349

After randomly observing melanized sporozoite in our mosquitoes, we wondered whether the melanization response could be higher in mosquitoes with a greater P. vivax infection intensity. Therefore, we performed three independent experimental infections to assess the melanization response of An. darlingi mosquitoes against the P. vivax sporozoite stage under two infection conditions. Mosquitoes obtained from the An. darlingi colony were deprived of sugar feeding for up to 12 hours before the blood meal. Two batches of 50 female An. darlingi mosquitoes, aged 3 to 5 days, were separated. For the first group, plasma from a P. vivax–positive blood sample was replaced with inactivated AB+ serum in a 1:1 (erythrocytes-to-serum) ratio, and this mixture was offered to the mosquitoes by DMFA. This group was designated SR. The second group was fed on the same P. vivax–positive blood sample but without serum replacement and was referred to as WB. The serum replacement approach is commonly used to increase infection rates in anopheline mosquitoes.5

After the blood meal, only completely engorged female mosquitoes were maintained in the study. At 14 dpi, the MGs and SGs of approximately 20 females from each group were dissected under a stereo microscope. Midguts were stained with 0.2% mercurochrome. Melanized sporozoites in both organs were visualized and counted using a microscope (×100).

The experiments were conducted with the approval of the Ethics Committee at the Centro de Pesquisa em Medicina Tropical (protocol number 530,106). All participants invited to participate in the study met the following criteria: positive diagnosis for P. vivax malaria, age ≥ 18 years, not pregnant, not indigenous, absence of severe or complicated malaria, and providing oral and signed consent to participate in the study. Oocyst intensity between the WB and SR groups, as well as the number of melanized sporozoites between WB-SG versus SR-SG and WB-MG versus SR-MG were compared using the Mann-Whitney test. The number of melanized sporozoites between WB-SG versus WB-MG and SR-SG versus SR-MG were compared using the Wilcoxon test for paired samples. Finally, the prevalence of mosquitoes with melanized sporozoites between the groups were compared by Fisher’s exact test. The 95% CIs were calculated by the Wald method, and Bonferroni correction was applied to the P-values.

Serum replacement before DMFA showed a significant increase in oocyst intensity (WB oocyst median = 16; SR oocyst median = 120.5; Mann-Whitney test, U = 246.5; P <0.0001) (Figure 2A). The number of melanized sporozoites was statistically higher in the MGs of the SR group than in the MGs of the WB group (WB melanized sporozoite median = 2; SR melanized sporozoite median = 13.5; Mann-Whitney test, U = 1,449; P = 0.0034). However, the quantity of melanized sporozoites between the SGs of the WB and SR groups did not show a statistical difference (WB melanized sporozoite median = 0; SR melanized sporozoite median = 0; Mann-Whitney test, U = 1,906; P = 0.6682). The proportion of mosquitoes with melanized sporozoites in the SGs was 38.1% (95% CI: 27.1–50.5) in the WB group and 42.9% (95% CI: 31.4–55.1) in the SR group. For MGs, 62.5% of mosquitoes had melanized sporozoites (95% CI: 50.2–73.3) in the WB group and 71.9% (95% CI: 59.8–81.5) in the SR group. However, no significant difference was observed between WB-SG versus SR-SG, as shown in Figure 2B.

Figure 2.
Figure 2.

Sporozoite melanization in Anopheles darlingi infected with Plasmodium vivax. (A) Oocyst intensity of mosquitoes fed under whole blood (WB) or serum replacement (SR) conditions. Each red circle represents a mosquito dissected. (B) Number of melanized sporozoites in each experimental group: whole blood–salivary gland (WB-SG); serum replacement–salivary gland (SR-SG); whole blood–midgut (WB-MG), and serum replacement–midgut (SR-MG) and the prevalence of mosquitoes with at least one melanized sporozoite. (C) Salivary gland and midgut paired melanized sporozoite in the WB group and the SR group. For (B) and (C) plots, each brown circle represents a salivary gland or a midgut dissected. The gray color in the pizza plot represents the proportion of mosquitoes without melanized sporozoites and the brown color represents the proportion of mosquitos with melanized sporozoites.

Citation: The American Journal of Tropical Medicine and Hygiene 110, 3; 10.4269/ajtmh.23-0349

When the MGs and SGs of the same group were compared, we observed more melanized sporozoites were present in the MGs than in the SGs, regardless of the group, whether WB or SR (Figure 2C) (WB group: Wilcoxon paired test, W = 585; P = 0.0002; SR group: Wilcoxon paired test, W = 1,120; P < 0.0001). In addition, a higher proportion of mosquitoes exhibited melanized sporozoites in the MGs than in the SGs for both experimental groups, WB and SR (Figure 2B).

DISCUSSION

Here, we report our observations of melanized P. vivax sporozoites located outside the MGs and SGs of An. darlingi on day 14 after the ingestion of a blood meal from P. vivax–infected human donors. Although melanization is a primary immune system response in insects, it is considered rare in susceptible strains.6,7 In refractory anopheline species, melanization is described primarily against the ookinete stages, as observed in the stable line Anopheles gambiae L3-5 used in immune response studies in anopheline mosquitoes.810

Anopheles darlingi is a highly susceptible neotropical anopheline species to P. falciparum and P. vivax malaria parasites11,12; however, it has been shown to have a lower sporozoite load than African and Asian anophelines.1315 To date, there have been no records of melanization in An. darlingi. For P. vivax and neotropical anopheline models, there are only two reports available in the literature about melanization. In 2000, Moreno and Berti16 studied the rate of oocyst melanization in an Anopheles aquasalis field captured in Venezuela and found melanized sporozoites of P. vivax in the hemocoel. In the second study, Hernández-Martínez et al.17 assessed the cellular-mediated reaction of Anopheles albimanus, a neotropical anopheline known to have poor vectorial competence to P. vivax, by injecting sporozoites through the mosquito thorax and observed the melanization of some of these parasites.

Previously, Simões et al.18 suggested that melanization in anopheline mosquitoes is directly dependent on the infection intensities of Plasmodium infection, as demonstrated in the An. albimanus–Plasmodium berghei and P. falciparum models. In this study, we induced a high P. vivax infection in An. darlingi by replacing plasma blood with inactivated serum AB+. As a result, we found more melanized sporozoites in the MGs of highly infected mosquitoes, as expected. Interestingly, we noted a higher number of melanized sporozoites outside MGs than outside SGs, regardless of the infection load.

Owing to some technique limitations, we were unable to measure the proportion of melanized sporozoites in the mosquitoes. However, nonmelanized sporozoites were present in all SGs where melanized sporozoites were observed. This suggests that the immune response of An. darlingi mosquitoes may be effectively evaded by P. vivax, as observed in the literature.19 The success of evasion seems to be genetically determined and may be dependent on Plasmodium–vector compatibility, as shown for the surface protein Pfs47, which allows P. falciparum to survive in their sympatric vectors.20 Another surface protein implicated in the evasion of mosquito immune response is the circumsporozoite protein (CSP). Studies have shown that conformational changes or posttranslational modification (glutaminyl cyclase) of this surface protein prevents the melanization of oocysts and sporozoites of Plasmodium.21,22 In the case of P. vivax, although it may evade the immunological response of An. darlingi, the vector’s ability to melanize free sporozoites in the hemocoel could be important in decreasing the transmission success of P. vivax.

In addition to genetic factors,9,19 nongenetic factors such as environmental conditions and the developmental stage of mosquitoes can influence them to mount a melanization response. For example, age and reproductive status,6,23 larval rearing conditions,24 and dietary resources in the adult stage6,25 can affect their response. Therefore, given the importance of P. vivax in malaria transmission in the Americas, particularly in the Amazon region, studying the ability of An. darlingi to develop a melanization response under different conditions, investigating whether field-captured An. darlingi exhibits a melanization response to P. vivax, and understanding the immune evasion strategies used by P. vivax may offer valuable insights for studies aimed at blocking malaria transmission.

ACKNOWLEDGMENT

We thanks all the volunteers in Porto Velho, RO, Brazil, who provided blood samples for the P. vivax DMFA assays, and we would like to thank Fiocruz Rondonia for providing the Microscopy platform (RPT-07i) to carry out this study.

REFERENCES

  • 1.

    WHO , 2022. World Malaria Report. Geneva, Switzerland: World Health Organization. https://www.who.int/publications/i/item/9789240064898. Accessed May 18, 2023.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 2.

    Kumar A , Srivastava P , Sirisena PDNN , Dubey SK , Kumar R , Shrinet J , Sunil S , 2018. Mosquito innate immunity. Insects 9: 95.

  • 3.

    Christensen BM , Li J , Chen CC , Nappi AJ , 2005. Melanization immune responses in mosquito vectors. Trends Parasitol 21: 192199.

  • 4.

    Araujo MS , Andrade AO , Santos NAC , Pereira DB , Costa GS , Paulo PFM , Rios CT , Moreno M , Pereira-da-Silva LH , Medeiros JF , 2019. Brazil’s first free-mating laboratory colony of Nyssorhynchus darlingi. Rev Soc Bras Med Trop 52: e20190159.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 5.

    Andrade AO et al., 2023. Optimization of Plasmodium vivax infection of colonized Amazonian Anopheles darlingi. Sci Rep 13: 18207.

  • 6.

    Schwartz A , Koella JC , 2002. Melanization of Plasmodium falciparum and C-25 Sephadex beads by field-caught Anopheles gambiae (Diptera: Culicidae) from southern Tanzania. J Med Entomol 29: 8489.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7.

    Niaré O et al., 2022. Genetic loci affecting resistance to human malaria parasites in a West African mosquito vector population. Science 298: 213216.

  • 8.

    Collins FH , Sakai RK , Vernick KD , Paskewitz S , Seeley DC , Miller LH , Collins WE , Campbell C , Gwadz RW , 1986. Genetic selection of a Plasmodium–refractory strain of the malaria vector Anopheles gambiae. Science 234: 607610.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9.

    Osta MA , Christophides GK , Kafatos FC , 2004. Effects of mosquito genes on Plasmodium development. Science 303: 20302032.

  • 10.

    Bladin S , Shiao SH , Moita LF , Janse CJ , Waters AP , Kafatos FC , Levashina EA , 2004. Complement-like protein TEP1 is a determinant of vectorial capacity in the malaria vector Anopheles gambiae. Cell 116: 661670.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 11.

    Klein TA , Lima JB , Tada MS , Miller R , 1991. Comparative susceptibility of anopheline mosquitoes in Rondonia, Brazil to infection by Plasmodium vivax. Am J Trop Med Hyg 45: 463470.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Grieco JP , Achee NL , Roberts DR , Andre RG , 2005. Comparative susceptibility of three species of Anopheles from Belize, Central America, to Plasmodium falciparum (NF-54). J Am Mosq Control Assoc 21: 279290.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Santos NAC et al., 2022. Evaluation of sustainable susceptibility to Plasmodium vivax infection among colonized Anopheles darlingi and Anopheles deaneorum. Malar J 21: 163.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14.

    Moreno M et al., 2014. Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax. Am J Trop Med Hyg 90: 612616.

  • 15.

    Villarreal-Treviño C , Vásquez GM , López-Sifuentes VM , Escibedo-Vargas K , Huayanay-Repetto A , Linton YM , Flores-Mendoza C , Lescano AG , Stell FM , 2015. Establishment of a free-mating, long-standing and highly productive laboratory colony of Anopheles darlingi from the Peruvian Amazon. Malar J 14: 227.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16.

    Moreno JE , Berti JA , 2000. Melanización de ooquistes y esporozoítos de Plasmodium vivax em poblaciones de Anopheles aquasalis Curry (Diptera: Culicidae) de Venezuela infectadas experimentalmente. Bol Entomol Venez 15: 1721.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17.

    Hernández-Martínez S , Lanz H , Rodríguez MH , González-Ceron L , Tsutsumi V , 2002. Cellular-mediated reactions to foreign organisms inoculated into the hemocoel of Anopheles albimanus (Diptera: Culicidae). J Med Entomol 39: 6169.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 18.

    Simões ML , Mlambo G , Tripathi A , Dong Y , Dimopoulos G , 2017. Immune regulation of Plasmodium is Anopheles species specific and infection intensity dependent. MBio 8: e01631e17.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19.

    Clayton AM , Dong Y , Dimopoulos G , 2014. The Anopheles innate immune system in the defense against malaria infection. J Innate Immun 6: 169181.

  • 20.

    Molina-Cruz A et al., 2020. Plasmodium falciparum evades immunity of anopheline mosquitoes by interacting with a Pfs47 midgut receptor. Proc Natl Acad Sci USA 117: 25972605.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21.

    Zhu F et al., 2022. Malaria oocysts require circumsporozoite protein to evade mosquito immunity. Nat Commun 13: 3208.

  • 22.

    Kolli SK et al., 2022. Malaria parasite evades mosquito immunity by glutaminyl cyclase-mediated posttranslational protein modification. Proc Natl Acad Sci USA 119: e2209729119.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23.

    Chun J , Riehle M , Paskewitz SM , 1995. Effect of mosquito age and reproductive status on melanization of Sephadez beads in Plasmodium-refractory and -susceptible strains of Anopheles gambiae. J Invertebr Pathol 66: 1117.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24.

    Suwanchaichinda C , Paskewitz SM , 1998. Effects of larval nutrition, adult body size, and adult temperature on the ability of Anopheles gambiae (Diptera: Culicidae) to melanize Sephadex beads. J Med Entomol 35: 157161.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25.

    Koella JC , Sorensen FL , 2002. Effect of adult nutrition on the melanization immune response of the malaria vector Anopheles stephensi. Med Vet Entomol 16: 316320.

    • PubMed
    • Search Google Scholar
    • Export Citation

Author Notes

Financial support: This work was supported, in part, by the Brazilian Ministry of Health/Departamento de Ciência e Tecnologia (DECIT)/Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) N° 23/2019 (grant number 442653/2019-0) and the Bill & Melinda Gates Foundation (INV-003970). Under the grant conditions of the foundation, a Creative Commons Attribution 4.0 Generic License has already been assigned to the author-accepted manuscript version that might arise from this submission. The study also received funding from Programa Pesquisa para o Sistema Único de Saúde (PPSUS)/Fundação de Amparo ao Desenvolvimento das Ações Científicas e Tecnológicas e à Pesquisa do Estado de Rondônia (FAPERO) N° 001/2020 (grant number 0012.259726/2020-70), and the International Centers of Excellence for Malaria Research (ICEMR) program GR109237 (CON-80002357). N.A.C. Santos expresses her appreciation for the fellowship granted by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (grant number 001) and J.F. Medeiros is a CNPq productivity fellow (304830/2022-4).

Authors’ addresses: Najara Akira Costa dos Santos and Jansen Fernandes Medeiros, Plataforma de Produção e Infecção de Vetores da Malária (PIVEM), Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Brazil and Instituto Nacional de Epidemiologia da Amazônia Ocidental (INCT-EpiAMO), Porto Velho, RO, Brazil, E-mails: najara.akira@fiocruz.br, and jansen.medeiro@fiocruz.br. Jéssica Evangelista Araújo, Programa de Pós-graduação em Biologia Experimental - Universidade Federal de Rondônia/Fiocruz Rondônia, Porto Velho, Brazil, and Plataforma de Produção e Infecção de Vetores da Malária (PIVEM), Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Brazil, E-mail: jessica.evangelista@fiocruz.br. Alessandra da Silva Bastos and José Daniel Costa Pontual, Plataforma de Produção e Infecção de Vetores da Malária (PIVEM), Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Brazil, E-mails: alessandra.bastos@fiocruz.br and jose.pontual@fiocruz.br. Joseph Michael Vinetz, Section of Infectious Diseases, Department of Internal Medicine, Yale School of Medicine, New Haven, CT, E-mail: joseph.vinetz@yale.edu. Maisa da Silva Araujo, Plataforma de Produção e Infecção de Vetores da Malária (PIVEM), Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Brazil, and Instituto Nacional de Epidemiologia da Amazônia Ocidental (INCT-EpiAMO), Porto Velho, RO, Brazil, Programa de Pós-Graduação em Conservação e uso de Recursos Naturais (PPGReN), Fundação Universidade Federal de Rondônia, Porto Velho, Brazil, and Laboratório de Pesquisa Translacional e Clíınica, Centro de Pesquisa em Medicina Tropical, Porto Velho, Brazil. E-mail: maisa.araujo@fiocruz.br.

Address correspondence to Maisa da Silva Araujo, Plataforma de Produção e Infecção de Vetores da Malária (PIVEM), Laboratório de Entomologia, Fiocruz Rondônia, Rua da Beira, 7671, Lagoa, Porto Velho, Rondônia, 76812–245, Brazil. Email: maisa.araujo@fiocruz.br; maisaraujo@gmail.com
  • Figure 1.

    Melanized Plasmodium vivax sporozoite in Anopheles darlingi mosquitoes. (A and D) Melanized sporozoites outside the midgut. (B, C, E, and F) Melanized and nonmelanized sporozoites outside the salivary glands. White arrowheads indicate melanized sporozoites, and red arrowheads indicate free sporozoites. The white box indicates the total magnification under light microscopy. All images were obtained at 14 days post infection.

  • Figure 2.

    Sporozoite melanization in Anopheles darlingi infected with Plasmodium vivax. (A) Oocyst intensity of mosquitoes fed under whole blood (WB) or serum replacement (SR) conditions. Each red circle represents a mosquito dissected. (B) Number of melanized sporozoites in each experimental group: whole blood–salivary gland (WB-SG); serum replacement–salivary gland (SR-SG); whole blood–midgut (WB-MG), and serum replacement–midgut (SR-MG) and the prevalence of mosquitoes with at least one melanized sporozoite. (C) Salivary gland and midgut paired melanized sporozoite in the WB group and the SR group. For (B) and (C) plots, each brown circle represents a salivary gland or a midgut dissected. The gray color in the pizza plot represents the proportion of mosquitoes without melanized sporozoites and the brown color represents the proportion of mosquitos with melanized sporozoites.

  • 1.

    WHO , 2022. World Malaria Report. Geneva, Switzerland: World Health Organization. https://www.who.int/publications/i/item/9789240064898. Accessed May 18, 2023.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 2.

    Kumar A , Srivastava P , Sirisena PDNN , Dubey SK , Kumar R , Shrinet J , Sunil S , 2018. Mosquito innate immunity. Insects 9: 95.

  • 3.

    Christensen BM , Li J , Chen CC , Nappi AJ , 2005. Melanization immune responses in mosquito vectors. Trends Parasitol 21: 192199.

  • 4.

    Araujo MS , Andrade AO , Santos NAC , Pereira DB , Costa GS , Paulo PFM , Rios CT , Moreno M , Pereira-da-Silva LH , Medeiros JF , 2019. Brazil’s first free-mating laboratory colony of Nyssorhynchus darlingi. Rev Soc Bras Med Trop 52: e20190159.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 5.

    Andrade AO et al., 2023. Optimization of Plasmodium vivax infection of colonized Amazonian Anopheles darlingi. Sci Rep 13: 18207.

  • 6.

    Schwartz A , Koella JC , 2002. Melanization of Plasmodium falciparum and C-25 Sephadex beads by field-caught Anopheles gambiae (Diptera: Culicidae) from southern Tanzania. J Med Entomol 29: 8489.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7.

    Niaré O et al., 2022. Genetic loci affecting resistance to human malaria parasites in a West African mosquito vector population. Science 298: 213216.

  • 8.

    Collins FH , Sakai RK , Vernick KD , Paskewitz S , Seeley DC , Miller LH , Collins WE , Campbell C , Gwadz RW , 1986. Genetic selection of a Plasmodium–refractory strain of the malaria vector Anopheles gambiae. Science 234: 607610.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9.

    Osta MA , Christophides GK , Kafatos FC , 2004. Effects of mosquito genes on Plasmodium development. Science 303: 20302032.

  • 10.

    Bladin S , Shiao SH , Moita LF , Janse CJ , Waters AP , Kafatos FC , Levashina EA , 2004. Complement-like protein TEP1 is a determinant of vectorial capacity in the malaria vector Anopheles gambiae. Cell 116: 661670.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 11.

    Klein TA , Lima JB , Tada MS , Miller R , 1991. Comparative susceptibility of anopheline mosquitoes in Rondonia, Brazil to infection by Plasmodium vivax. Am J Trop Med Hyg 45: 463470.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Grieco JP , Achee NL , Roberts DR , Andre RG , 2005. Comparative susceptibility of three species of Anopheles from Belize, Central America, to Plasmodium falciparum (NF-54). J Am Mosq Control Assoc 21: 279290.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Santos NAC et al., 2022. Evaluation of sustainable susceptibility to Plasmodium vivax infection among colonized Anopheles darlingi and Anopheles deaneorum. Malar J 21: 163.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14.

    Moreno M et al., 2014. Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax. Am J Trop Med Hyg 90: 612616.

  • 15.

    Villarreal-Treviño C , Vásquez GM , López-Sifuentes VM , Escibedo-Vargas K , Huayanay-Repetto A , Linton YM , Flores-Mendoza C , Lescano AG , Stell FM , 2015. Establishment of a free-mating, long-standing and highly productive laboratory colony of Anopheles darlingi from the Peruvian Amazon. Malar J 14: 227.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16.

    Moreno JE , Berti JA , 2000. Melanización de ooquistes y esporozoítos de Plasmodium vivax em poblaciones de Anopheles aquasalis Curry (Diptera: Culicidae) de Venezuela infectadas experimentalmente. Bol Entomol Venez 15: 1721.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17.

    Hernández-Martínez S , Lanz H , Rodríguez MH , González-Ceron L , Tsutsumi V , 2002. Cellular-mediated reactions to foreign organisms inoculated into the hemocoel of Anopheles albimanus (Diptera: Culicidae). J Med Entomol 39: 6169.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 18.

    Simões ML , Mlambo G , Tripathi A , Dong Y , Dimopoulos G , 2017. Immune regulation of Plasmodium is Anopheles species specific and infection intensity dependent. MBio 8: e01631e17.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19.

    Clayton AM , Dong Y , Dimopoulos G , 2014. The Anopheles innate immune system in the defense against malaria infection. J Innate Immun 6: 169181.

  • 20.

    Molina-Cruz A et al., 2020. Plasmodium falciparum evades immunity of anopheline mosquitoes by interacting with a Pfs47 midgut receptor. Proc Natl Acad Sci USA 117: 25972605.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21.

    Zhu F et al., 2022. Malaria oocysts require circumsporozoite protein to evade mosquito immunity. Nat Commun 13: 3208.

  • 22.

    Kolli SK et al., 2022. Malaria parasite evades mosquito immunity by glutaminyl cyclase-mediated posttranslational protein modification. Proc Natl Acad Sci USA 119: e2209729119.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23.

    Chun J , Riehle M , Paskewitz SM , 1995. Effect of mosquito age and reproductive status on melanization of Sephadez beads in Plasmodium-refractory and -susceptible strains of Anopheles gambiae. J Invertebr Pathol 66: 1117.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24.

    Suwanchaichinda C , Paskewitz SM , 1998. Effects of larval nutrition, adult body size, and adult temperature on the ability of Anopheles gambiae (Diptera: Culicidae) to melanize Sephadex beads. J Med Entomol 35: 157161.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25.

    Koella JC , Sorensen FL , 2002. Effect of adult nutrition on the melanization immune response of the malaria vector Anopheles stephensi. Med Vet Entomol 16: 316320.

    • PubMed
    • Search Google Scholar
    • Export Citation
Past two years Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 699 699 78
PDF Downloads 271 271 48
 
 
 
 
Affiliate Membership Banner
 
 
Research for Health Information Banner
 
 
CLOCKSS
 
 
 
Society Publishers Coalition Banner
Save