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Effects of Anticoagulants on Plasmodium vivax Oocyst Development in Anopheles albimanus Mosquitoes

Artificial membrane feeding (AMF) assays are used to determine malaria transmission-blocking activity in Anopheles. The purpose of this study was to determine the effect of the most widely used anticoagulants, EDTA and heparin, on development of the Plasmodium vivax sporogonic cycle. Blood samples collected from 60 patients carrying P. vivax infections were used to feed An. albimanus using AMF. Seven days after feeding, mosquitoes were dissected to assess mosquito infection. Mosquitoes fed with blood containing EDTA showed a lower mean oocyst number as compared with those fed blood with heparin. However, this effect was minimized upon reduction of EDTA concentrations in the serum. This result may be explained by the fact that microgametocytes require Ca²⁺, Mn²⁺, and Mg⁺² to activate enzymes important for exflagellation process and for motility of ookinetes. We therefore recommend that heparin be used as the anticoagulant of choice for blood used in AMF assays.

Received December 26, 2006. Accepted for publication April 1, 2007.

Acknowledgments: We thank the endemic communities of Buenaventura for providing blood samples for this study and the Programa de Enfermedades Tropicales (PET) for support in patient recruitment. We are also grateful to Dr. Felipe Zamora for his suggestions and critical discussions of results and to Einer Celorio of the Malaria Vaccine and Drug Development Center (Cali, Colombia) and Dr. Mario Chen-Mok and Beth Robinson of Family Health International (Durham, NC) for helpful discussions related to parts of this work.

Financial support: This study was supported by the National Institute of Allergy and Infectious Diseases (US/NIH/NIAID) through TMRC grant # 49486 and by Colciencias contract # 256-2005.

^* Address correspondence to Myriam Arévalo-Herrera, Malaria Vaccine and Drug Development Center, Carrera 35 No 4A-53, AA 26020, Cali, Colombia. E-mail: marevalo{at}inmuno.org

Authors’ addresses: Yezid Solarte, Leonardo Rocha, Zuleyma Castillo, Sócrates Herrera, and Myriam Arévalo-Herrera, Malaria Vaccine and Drug Development Center, Carrera 35 No. 4A-53, AA 26020, Cali, Colombia, Telephone: +57-2-5583937, and Instituto de Inmunología, Facultad de Salud, Universidad del Valle, Sede San Fernando, AA 25574, Cali, Colombia, Telephone: +57-2-558-1931, Fax: +57-2-556-0141, E-mail: marevalo{at}inmuno.org. Maria del Ro-sario Manzano, Universidad Nacional de Colombia, via a Candelaria, Palmira, Colombia, Telephone: 57-271-7000, ex. 35380. Mark A. James, Department of Tropical Medicine, Tulane University, 1440 Canal Street, SL-17, New Orleans, LA, Telephone: +1 (504) 988-2503; Fax: +1 (504) 988-7313.

Reprint requests: Myriam Arévalo-Herrera, Malaria Vaccine and Drug Development Center, Carrera 35 No 4A-53, AA 26020, Cali, Colombia, E-mail: marevalo{at}inmuno.org.