Original Article

Evaluation of Isotope 32P Method to Mark Culex pipiens (Diptera: Culicidae) in a Laboratory


Background: The aim of the current study was to develop a marking technique as an internal marker to mark post blood meal mosquitoes by using stable phosphate isotope 32P and determine the optimal concentration of it.
: An isotonic physiological saline solution, containing different concentration of radioactive isotope 32P-labeled disodium phosphate (Na2H32PO4) was injected into rabbits via the jugular vein in the laboratory. Emerged Cx. pipiens were marked after feeding on rabbit. At the same time, the labeled conditions of emerged Cx. pipiens were also measured by placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside.
: According to the label condition of Cx. pipiens after taking blood and the effect of different dosage Na2H32PO4 on rabbit health, the optimal concentration of radioactive isotope was determined, that is, 0.1211 mCi/kg. By placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside, the emerged mosquitoes were also labeled. Therefore, feeding mosquitoes on the animal injected with radioactive Na2H32PO4 was more prac­tical for detecting and tracing mosquitoes.
The method was less time-consuming, more sensitive and safer. This marking method will facilitate post-bloodmeal studies of mosquitoes and other blood-sucking insects.

Abdel-Malek AA (1966) Study of the dis- persion and flight range of Anopheles sergentii Theo in Siwa Oasis using ra- dioactive isotopes as markers. Bull World Health Organ. 35: 968–973.

Akey DH, Burns DW (1991) Analytical con- sideration and methodologies for ele- mental determinations in biological samples. Southwest Entomol. 3: 25–36.

Anderson RA, Edman JD, Scott TW (1990) Rubidium and cesium as host blood- markers to study multiple blood feed- ing by mosquitoes (Diptera: Culicidae). J Med

E ntomol. 27: 999–1001.

Baber I, Keita M, Sogoba N, Konate M, Di- allo M, Doumbia S, Traore SF, Ribeiro JMC, Manoukis NC (2010) Population size and migration of Anopheles gam- biae in the Bancoumana region of Mali and their signifcance for effcient vector control. PLoS ONE 5.

Basavaraju Y, Devi BSR, Mukthayakka G, Reddy PL, Mair GC (1998) Evaluation of marking and tagging methods for genetic studies in carp. J Biosci. 23:585–593.

Bellini R, Albieri A, Balestrino F, Carrieri M, Porretta D, Urbanelli S, Calvitti M, Moretti R, Maini S (2010) Dispersal and survival of Aedes albopictus (Dip- tera: Culicidae) males in Italian urban areas and significance for sterile insect technique application. J Med Entomol.47: 1082–1091.

Bruce-Chwatt LJ (1956) Radioisotopes for research on and control of mosquitos. Bull World Health Organ. 15: 491–511.

Burkett-Cadena ND, Eubanks MD, Unnasch TR (2008a) Preference of female mos- quitoes for natural and artificial resting sites. J Am Mosq Control Assoc. 24:228–235.

Burkett-Cadena ND, Graham SP, Hassan HK, Guyer C, Eubanks MD, Katholi CR, Unnasch TR (2008b) Blood feed- ing patterns of potential arbovirus vec- tors of the genus Culex targeting ecto- thermic hosts. Am J Trop Med Hyg.79: 809–815.

Ciota AT, Drummond CL, Ruby MA, Drob- nack J, Ebel GD, Kramer LD (2012) Dispersal of Culex mosquitoes (Dip- tera: Culicidae) from a wastewater treat- ment facility. J Med Entomol. 49: 35–42.

Dissanaike AS, Dissanaike GA, Niles WJ, Surendranathan R (1957) Further Studies on Radioactive Mosquitoes and Filarial Larvae using Autoradiographic Tech- nique. Exp Parasitol. 6: 261–270.

Dudley JE, Searles EM (1923) Color mark- ing of the striped cucumber beetle (Diabrotica vittata Fab.) and prelimi- nary experiments to determine its flight. J Econ Entomol. 16: 363–368.

Fisher J, Peterson RT (1964) The World of Birds. Garden City, NY: Doubleday. p.288.

Fleischer SJ, Gaylor MJ, Hue NV, Graham LC (1986) Uptake and elimination of rubidium, a physiological marker, in adult Lygus lineolaris (Hemiptera: Miridae). Ann Entomol Soc Am. 79:19–25.

Geiger JC, Purdy WC, Tarbett RE (1919) Effective malarial control in a rice field district with observations on ex- perimental mosquito flights. J Am Med Assoc. 72: 844–847.

Guillebeau LP, All JN, Nutter FW, Kuhn C (1993) Comparison of foliar and soil- drench applications of aqueous rubid- ium chloride solution to plants for marking feeding aphids (Homoptera: Aphidae). J Entomol Sci. 28: 370–375.

Hagler JR, Jackson CG (2001) Methods for marking insects: current techniques and future prospects. Annu Rev Ento- mol. 46: 511–543.

Hamer GL, Donovan DJ, Hood-Nowotny R, Kaufman MG, Goldberg TL, Walker ED (2012) Evaluation of a stable iso- tope method to mark naturally-breeding larval mosquitoes for adult dispersal studies. J Med Entomol. 49: 61–70.

Hasset CC, Jenkins DW (1949) Production of radioactive mosquitoes. Science.110: 109–110.

Hasset CC, Jenkins DW (1951) The uptake and effect of radiophosphorus in mos- quitoes. Physiol Zool. 24: 257.

Hayes JL (1989) Detection of single and multiple trace element labels in indi- vidual eggs of diet-reared Heliothis vi- rescens (Lepidoptera: Noctuidae). Ann Entomol Soc Am. 82: 340–345.

Helinski MEH, Hood-Nowotny R, Mayr L, Knols BGJ (2007) Stable isotope-mass spectrometric determination of semen transfer in malaria Mosquitoes. J Exp Bio. 210: 1266–1274.

Holbrook FR, Belden RP, Bobian RJ (1991) Rubidium for marking adults of Culi- coides Variipennis (Diptera: Cerato- pogonidae). J Med Entomol. 28: 246–249.

Hood-Nowotny R, Knols BGJ (2007) Stable isotope methods in biological and eco- logical studies of arthropods. Entomol Exp Appl. 124: 3–16.

Hood-Nowotny R, Mayr L, Knols B (2006) Use of carbon-13 as a population marker for Anopheles arabiensis in a sterile insect technique (SIT) context. Malar J. 5: 6.

IAEA International Atomic Energy Agency (2009) Manual for the Use of Stable Isotopes in Entomology. International Atomic Energy Agency 978-92-0-102209-7, Vienna, Austria.

Jenkins DW (1949) A field method for mark- ing arctic mosquitoes with radio- phoshorus. J Econ Entomol. 40: 988–989.

Lapointe DA (2008) Dispersal of Culex quinquefasciatus (Diptera: Culicidae) in a Hawaiian rain forest. J Med En- tomol. 45: 600–609.

Lindquist AW, Ikeshoji T, Grab B, Demeillo B, Khan ZH (1967) Dispersion studies of Culex pipiens fatigans tagged with 32P in Kemmendine area of Rangoon, Burma. Bull World Health Org. 36:21–37.

Linton D, Safranyik L, McMullen L, Betts R (1987) Field techniques for rearing and marking mountain pine beetle for use in dispersal studies. J Entomol Soc B C. 84: 53–58.

Liu QY, Liu XB, Zhou GC, Jiang JY, Guo YH, Ren DS, Zheng CJ, Wu HX, Yang SR, Liu JL, Li HS, Li HZ, Li Q, Yang WZ, Chu C (2012) Dispersal Range of Anopheles sinensis in Yong- cheng City, China by Mark-Release- Recapture Methods. PLoS ONE. 7(11): e51209.

Macneale KH, Peckarsky BL, Likens GE (2004) Contradictory results from dif- ferent methods for measuring direction of insect flight. Freshwater Biol. 49:1260–1268.

Macneale KH, Peckarsky BL, Likens GE (2005) Stable isotopes identify disper- sal patterns of stonefly populations liv- ing along stream corridors. Freshwater Biol. 50: 1117–1130.

McCall PJ, Mosha FW, Njunwa KJ, Sher- lock K (2001) Evidence for memo- rized sitefidelity in Anopheles ara- biensis. Trans R Soc Trop Med Hyg.95: 587–590.

McMullen L, Safranyik L, Linton D, Betts R (1988) Survival of self-marked moun- tain pine beetles emerged from logs dusted with fluorescent powder. J En- tomol Soc B C. 85: 25–28.

Messing R, Klungness L, Purcell M, Wong T (1993) Quality control parameters of mass-reared opiine parasitoids used in augmentative biological control of tephritid fruit flies in Hawaii. Biol Control. 3: 140–147.

Midega JT, Mbogo CM, Mwnambi H, Wil-son MD, Ojwang G, Mwangangi JM, Nzovu JG, Githure JI, Yan G, Beier JC (2007) Estimating dispersal and sur- vival of Anopheles gambiae and Anopheles funestus along the Kenyan coast by using mark-release-recapture methods. J Med Entomol. 44: 923–929.

Moffitt HR, Albano DJ (1972) Codling moths: fluorescent powders as mark- ers. Environ Entomol. 1: 750–753.

Molaei G, Andreadis TA, Armstrong PM, Anderson JF, Vossbrinck CR (2006) Host feeding patterns of Culex mosqui- toes and West Nile virus transmission, northeastern United States. Emerg In- fect Dis. 12: 468–474.

Nakata T (2008) Effectiveness of micronized fluorescent powder for marking citrus psyllid Diaphorina citri. Appl Entomol Zool. 43: 33–36.

Narisu, Lockwood JA, Schell SP (1999) A novel mark-recapture technique and its application to monitoring the direction and distance of local movements of rangeland grasshoppers (Orthoptera: Acrididae) in the context of pest man- agement. J Appl Ecol. 36: 604–617.

Niebylski ML, Meek CL (1989) A self-marker device for emergent adult mosquitoes. J Am Mosq Control Assoc. 5: 86–90.

O'Brien RD, Wolfe LS (1964) Radiation, Radioactivity and Insects. Academic Press, New York, USA.

Pates H (2002) Zoophilic and anthropophilic behaviour in the Anopheles gambiae complex. PhD, London: University of London, p. 206.

Porter SD, Jorgenson CD (1980) Recapture studies of the harvester ant, Pogo- nomyrmex owyheei Cole, using a fluo- rescent marking technique. Ecol En- tomol. 5: 263–269.

Quarterman KD, Jensen JA, Mathis W, Smith WW (1955) Flight dispersal of rice field mosquitoes in Arkansas. J Econ Entomol. 48: 30–32.

Quarterman KD, Mathis W, Kilpatrick JW (1954) Urban fly dispersal in the area of Savannah, Georgia. J Econ Ento- mol. 47: 405–412.

Reinecke J (1990) A rapid and controllable technique for surface labeling boll weevils with fluorescent pigments. Southwest Entomol. 15: 309–316.

Reisen WK, Aslam Y, Siddiqui TF, Khan AQ (1978) A mark-release-recapture experiment with Culex tritaeniorhyn- chus Giles. 72: 167–177.

Reisen WK, Lothrop HD, Lothrop B (2003) Factors influencing the outcome of mark-release-recapture studies with Culex tarsalis (Diptera: Culicidae). J Med Entomol. 40: 820–829.

Reisen WK, Milby MM, Meyer RP (1992) Pop- ulation-dynamics of adult Culex mos-quitoes (Diptera, culicidae) along the Kern Kern River, Kern-County, Cali- fornia, in 1990. J Med Entomol. 29:531–543.

Russell RC, Webb CE, Williams CR, Ritchie SA (2005) Mark-release-recapture study to measure dispersal of the mosquito Aedes aegypti in Cairns, Queensland, Australia. Med Vet Entomol. 19: 451–457.

Sardelis MR, Turell MJ, O’Guinn ML, An- dre RG, Roberts DR (2002) Vector competence of three North American strains of Aedes albopictus for West Nile virus. J Am Mosq Control Assoc.18: 284–289.

Service MW (1993) Mosquito ecology-field sampling methods. London: Chapman and Hall. 988.

Sheppard P, Macdonald W, Tonn R, Grab B (1969) The dynamics of an adult popu- lation of Aedes aegypti in relation to dengue haemorrhagic fever in Bang- kok. J An Ecola. 38: 661–702.

Silver JB (2008) Mosquito Ecology: Fieeld Sampling Methods. Springer, New York.

Smith AH, Kleiber M, Black AL, Edick M, Robinson RR, Heitman H Jr (1951) Distribution of intravenously injected radioactive phosphorus P32 among swine tissues. J Animal Sci. 10: 893–901.

Solberg VB, Bernier L, Schneider I, Burge R, Writz RA (1999) Rubidium mark- ing of Anopheles stephensi (Diptera: Culicidae). J Med Entomol. 36: 141–143.

Southwood TRE (1978) Absolute population estimates using marking techniques. Ecological Methods, (ed. by TRE Southwood and PA Henderson), Chap- man and Hall, New York, USA, pp.70–129.

Stimmann MW, Wolf WW, Berry WL (1973) Cabbage loopers: the biological effects of rubidium in the larval diet. J Econ Entomol. 66: 324–326.

Takken W, Charlwood DJ, Billingsley PF, Gort G (1998) Dispersal and survival of Anopheles funestus and A. gambiae s.l. (Diptera: Culicidae) during the rainy season in southeast Tanzania. Bull Entomol Res. 88: 561–566.

Trpis M, Hausermann W (1986) Dispersal and other population parameters of Aedes aegypti in an African village and their possible significance in epi- demiology of vector-borne diseases. Am J Trop Med Hyg. 35: 1263–1279.

Valerio L, Facchinelli L, Ramsey JM, Bond JG, Scott TW (2012) Dispersal of male Aedes aegypti in a coastal village in southern Mexico. Am J Trop Med Hyg. 86: 665–676.

Van Steenwyk RA, Kaneshiro KY, Hue NV, Whittier TS (1992) Rubidium as an in- ternal physiological marker for Mediter- ranean fruit fly (Diptera: Tephritidae). J Econ Entomol. 85: 2357–2364.

Verhulst NO, Loonen JA, Takken W (2013) Advances in methods for colour mark- ing of mosquitoes. Parasit Vectors. 6:200. doi: 10.1186/1756-3305-6-200. Walker ED, Copeland RS, Paulson SL,Munstermann LE (1987) Adult survi- vorship, population density, and body size in sympatric populations of Aedes triseriatus and Aedes hendersoni (Dip- tera: Culicidae). J Med Entomol. 24:485–493.

Welch CH, Kline DL, Allan SA, Barnard DR (2006) Laboratory evaluation of a dyed food marking technique for Culex quinquefasciatus (Diptera: Culicidae). J Am Mosq Control Assoc. 22: 626–628.

Wilkins EE, Smith SC, Roberts JM, Benedict M (2007) Rubidium marking of Anopheles mosquitoes detectable by field-capable X-ray spectrometry. Med Vet Entomol. 21: 196–203.

IssueVol 10 No 2 (2016) QRcode
SectionOriginal Article
Radioactive isotope Mark Culex pipiens Rabbit

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How to Cite
Zhang C, Shi G, Zhao Y, Yan D, Li H, Liu H, Wiwatanaratanabutr I, Gong M. Evaluation of Isotope 32P Method to Mark Culex pipiens (Diptera: Culicidae) in a Laboratory. J Arthropod Borne Dis. 2016;10(2):211-221.