Skip to main content
Top
Published in: Malaria Journal 1/2011

Open Access 01-12-2011 | Research

Evaluating RNAlater® as a preservative for using near-infrared spectroscopy to predict Anopheles gambiae age and species

Authors: Maggy Sikulu, Kayla M Dowell, Leon E Hugo, Robert A Wirtz, Kristin Michel, Kamaranga HS Peiris, Sarah Moore, Gerry F Killeen, Floyd E Dowell

Published in: Malaria Journal | Issue 1/2011

Login to get access

Abstract

Background

Mosquito age and species identification is a crucial determinant of the efficacy of vector control programmes. Near-infrared spectroscopy (NIRS) has previously been applied successfully to rapidly, non-destructively, and simultaneously determine the age and species of freshly anesthetized African malaria vectors from the Anopheles gambiae s.l. species complex: An. gambiae s. s. and Anopheles arabiensis. However, this has only been achieved on freshly-collected specimens and future applications will require samples to be preserved between field collections and scanning by NIRS. In this study, a sample preservation method (RNAlater®) was evaluated for mosquito age and species identification by NIRS against scans of fresh samples.

Methods

Two strains of An. gambiae s.s. (CDC and G3) and two strains of An. arabiensis (Dongola, KGB) were reared in the laboratory while the third strain of An. arabiensis (Ifakara) was reared in a semi-field system. All mosquitoes were scanned when fresh and rescanned after preservation in RNAlater® for several weeks. Age and species identification was determined using a cross-validation.

Results

The mean accuracy obtained for predicting the age of young (<7 days) or old (≥ 7 days) of all fresh (n = 633) and all preserved (n = 691) mosquito samples using the cross-validation technique was 83% and 90%, respectively. For species identification, accuracies were 82% for fresh against 80% for RNAlater® preserved. For both analyses, preserving mosquitoes in RNAlater® was associated with a highly significant reduction in the likelihood of a misclassification of mosquitoes as young or old using NIRS. Important to note is that the costs for preserving mosquito specimens with RNAlater® ranges from 3-13 cents per insect depending on the size of the tube used and the number of specimens pooled in one tube.

Conclusion

RNAlater® can be used to preserve mosquitoes for subsequent scanning and analysis by NIRS to determine their age and species with minimal costs and with accuracy similar to that achieved from fresh insects. Cold storage availability allows samples to be stored longer than a week after field collection. Further study to develop robust calibrations applicable to other strains from diverse ecological settings is recommended.
Appendix
Available only for authorised users
Literature
1.
go back to reference Bayoh MN, Mathias D, Odiere M, Mutuku F, Kamau L, Gimnig J, Vulule J, Hawley W, Hamel M, Walker E: Anopheles gambiae: historical population decline associated with regional distribution of insecticide-treated bed nets in western Nyanza Province, Kenya. Malar J. 2010, 9: 62-10.1186/1475-2875-9-62.PubMedCentralCrossRefPubMed Bayoh MN, Mathias D, Odiere M, Mutuku F, Kamau L, Gimnig J, Vulule J, Hawley W, Hamel M, Walker E: Anopheles gambiae: historical population decline associated with regional distribution of insecticide-treated bed nets in western Nyanza Province, Kenya. Malar J. 2010, 9: 62-10.1186/1475-2875-9-62.PubMedCentralCrossRefPubMed
2.
go back to reference Russell T, Lwetoijera D, Maliti D, Chipwaza B, Kihonda J, Charlwood JD, Smith T, Lengeler C, Mwanyangala M, Nathan R, Knols BG, Takken W, Killeen GF: Impact of promoting longer-lasting insecticide treatment of bed nets upon malaria transmission in a rural Tanzanian setting with pre-existing high coverage of untreated nets. Malar J. 2010, 9: 187-10.1186/1475-2875-9-187.PubMedCentralCrossRefPubMed Russell T, Lwetoijera D, Maliti D, Chipwaza B, Kihonda J, Charlwood JD, Smith T, Lengeler C, Mwanyangala M, Nathan R, Knols BG, Takken W, Killeen GF: Impact of promoting longer-lasting insecticide treatment of bed nets upon malaria transmission in a rural Tanzanian setting with pre-existing high coverage of untreated nets. Malar J. 2010, 9: 187-10.1186/1475-2875-9-187.PubMedCentralCrossRefPubMed
3.
go back to reference Gillies M, Smith A: The effect of a residual house-spraying campaign in East Africa on species balance in the Anopheles funestus Group. The replacement of A. funestus Giles by A. rivulorum Leeson. Bull Entomol Res. 1960, 51: 243-252. 10.1017/S0007485300057953.CrossRef Gillies M, Smith A: The effect of a residual house-spraying campaign in East Africa on species balance in the Anopheles funestus Group. The replacement of A. funestus Giles by A. rivulorum Leeson. Bull Entomol Res. 1960, 51: 243-252. 10.1017/S0007485300057953.CrossRef
4.
go back to reference Garret-Jones C: Prognosis for interuption of malaria transmission through assessment of the mosquito's vectorial capacity. Nature. 1964, 204: 1173-1175. 10.1038/2041173a0.CrossRef Garret-Jones C: Prognosis for interuption of malaria transmission through assessment of the mosquito's vectorial capacity. Nature. 1964, 204: 1173-1175. 10.1038/2041173a0.CrossRef
5.
go back to reference Dye C: The analysis of parasite transmission by bloodsucking insects. Annu Rev Ent. 1992, 37: 1-19. 10.1146/annurev.en.37.010192.000245.CrossRef Dye C: The analysis of parasite transmission by bloodsucking insects. Annu Rev Ent. 1992, 37: 1-19. 10.1146/annurev.en.37.010192.000245.CrossRef
6.
go back to reference Magesa SM, Wilkes TJ, Mnzava AEP, Njunwa KJ, Myamba J, Kivuyo MDP, Hill N, Lines JD, Curtis CF: Trial of pyrethroid impregnated bednets in an area of Tanzania holoendemic for malaria Part 2. Effects on the malaria vector population. Acta Trop. 1991, 49: 97-108. 10.1016/0001-706X(91)90057-Q.CrossRefPubMed Magesa SM, Wilkes TJ, Mnzava AEP, Njunwa KJ, Myamba J, Kivuyo MDP, Hill N, Lines JD, Curtis CF: Trial of pyrethroid impregnated bednets in an area of Tanzania holoendemic for malaria Part 2. Effects on the malaria vector population. Acta Trop. 1991, 49: 97-108. 10.1016/0001-706X(91)90057-Q.CrossRefPubMed
7.
go back to reference Robert V, Carnevale P: Influence of deltamethrin treatment of bed nets on malaroa transmission in the Kou valley, Burkina Faso. Bull World Health Organ. 1991, 69: 735-740.PubMedCentralPubMed Robert V, Carnevale P: Influence of deltamethrin treatment of bed nets on malaroa transmission in the Kou valley, Burkina Faso. Bull World Health Organ. 1991, 69: 735-740.PubMedCentralPubMed
8.
go back to reference Beier C: Malaria parasite development in mosquitoes. Annu Rev Ent. 1998, 43: 519-543. 10.1146/annurev.ento.43.1.519.CrossRef Beier C: Malaria parasite development in mosquitoes. Annu Rev Ent. 1998, 43: 519-543. 10.1146/annurev.ento.43.1.519.CrossRef
9.
go back to reference Saul A: Zooprophylaxis or zoopotentiation: the outcome of introducing animals on vector transmission is highly dependent on the mosquito mortality while searching. Malar J. 2003, 2: 32-10.1186/1475-2875-2-32.PubMedCentralCrossRefPubMed Saul A: Zooprophylaxis or zoopotentiation: the outcome of introducing animals on vector transmission is highly dependent on the mosquito mortality while searching. Malar J. 2003, 2: 32-10.1186/1475-2875-2-32.PubMedCentralCrossRefPubMed
10.
go back to reference Killeen GF, Smith TA: Exploring the contributions of bed nets, cattle, insecticides and excitorepellency to malaria control: a deterministic model of mosquito host-seeking behaviour and mortality. Trans R Soc Trop Med Hyg. 2007, 101: 867-880. 10.1016/j.trstmh.2007.04.022.PubMedCentralCrossRefPubMed Killeen GF, Smith TA: Exploring the contributions of bed nets, cattle, insecticides and excitorepellency to malaria control: a deterministic model of mosquito host-seeking behaviour and mortality. Trans R Soc Trop Med Hyg. 2007, 101: 867-880. 10.1016/j.trstmh.2007.04.022.PubMedCentralCrossRefPubMed
11.
go back to reference Killeen GF, Smith TA, Ferguson HM, Mshinda H, Abdulla S, Lengeler C, Kachur SP: Preventing childhood malaria in Africa by protecting adults from mosquitoes with insecticide-treated nets. PLoS Med. 2007, 4: e229-10.1371/journal.pmed.0040229.PubMedCentralCrossRefPubMed Killeen GF, Smith TA, Ferguson HM, Mshinda H, Abdulla S, Lengeler C, Kachur SP: Preventing childhood malaria in Africa by protecting adults from mosquitoes with insecticide-treated nets. PLoS Med. 2007, 4: e229-10.1371/journal.pmed.0040229.PubMedCentralCrossRefPubMed
12.
go back to reference Polovodova VP: The determination of the physiological age of female Anopheles by number of gonotrophic cycles completed. Med Parazitol Parazitar Bolezni. 1949, 18: 352-355. Polovodova VP: The determination of the physiological age of female Anopheles by number of gonotrophic cycles completed. Med Parazitol Parazitar Bolezni. 1949, 18: 352-355.
13.
go back to reference Gillies M: A modified technique for the age-grading of populations of Anopheles gambiae. Ann Trop Med Parasit. 1958, 52: 261-273.PubMed Gillies M: A modified technique for the age-grading of populations of Anopheles gambiae. Ann Trop Med Parasit. 1958, 52: 261-273.PubMed
14.
go back to reference Gillies M: Studies on the dispersion and survival of Anopheles gambiae in East Africa, by means of marking and release experiments. Bull Entomol Res. 1961, 52: 99-127. 10.1017/S0007485300055309.CrossRef Gillies M: Studies on the dispersion and survival of Anopheles gambiae in East Africa, by means of marking and release experiments. Bull Entomol Res. 1961, 52: 99-127. 10.1017/S0007485300055309.CrossRef
15.
go back to reference Detinova T: Age-grouping methods in Diptera of medical importance, with special reference to some vectors of malaria. Monogr Ser World Health Organ. 1962, 47: 13-191.PubMed Detinova T: Age-grouping methods in Diptera of medical importance, with special reference to some vectors of malaria. Monogr Ser World Health Organ. 1962, 47: 13-191.PubMed
16.
go back to reference Gillies MT, Wilkes TJ: A study of the age composition of Anopheles gambiae Giles and A. funestus Giles in north-eastern Tanzania. Bull Entomol Res. 1965, 56: 237-262. 10.1017/S0007485300056339.CrossRefPubMed Gillies MT, Wilkes TJ: A study of the age composition of Anopheles gambiae Giles and A. funestus Giles in north-eastern Tanzania. Bull Entomol Res. 1965, 56: 237-262. 10.1017/S0007485300056339.CrossRefPubMed
17.
go back to reference Caputo B, Dani R, Horne L, Petrarca V, Turillazzi S, Coluzzi M, Priestman A, Torre A: Identification and composition of cuticular hydrocarbons of the major Afrotropical malaria vector Anopheles gambiae s.s. (Diptera: Culicidae): analysis of sexual dimorphism and age-related changes. J Mass Spectrom. 2005, 40: 1595-1604. 10.1002/jms.961.CrossRefPubMed Caputo B, Dani R, Horne L, Petrarca V, Turillazzi S, Coluzzi M, Priestman A, Torre A: Identification and composition of cuticular hydrocarbons of the major Afrotropical malaria vector Anopheles gambiae s.s. (Diptera: Culicidae): analysis of sexual dimorphism and age-related changes. J Mass Spectrom. 2005, 40: 1595-1604. 10.1002/jms.961.CrossRefPubMed
18.
go back to reference Marinotti O, Nguyen QK, Calvo E, James AA, Ribeiro JMC: Microarray analysis of genes showing variable expression following a blood meal in Anopheles gambiae. Insect Mol Biol. 2005, 14: 365-373. 10.1111/j.1365-2583.2005.00567.x.CrossRefPubMed Marinotti O, Nguyen QK, Calvo E, James AA, Ribeiro JMC: Microarray analysis of genes showing variable expression following a blood meal in Anopheles gambiae. Insect Mol Biol. 2005, 14: 365-373. 10.1111/j.1365-2583.2005.00567.x.CrossRefPubMed
19.
go back to reference Marinotti O, Calvo E, Nguyen QK, Dissanayake S, Ribeiro JMC, James AA: Genome-wide analysis of gene expression in adult Anopheles gambiae. Insect Mol Biol. 2006, 15: 1-12. 10.1111/j.1365-2583.2006.00610.x.CrossRefPubMed Marinotti O, Calvo E, Nguyen QK, Dissanayake S, Ribeiro JMC, James AA: Genome-wide analysis of gene expression in adult Anopheles gambiae. Insect Mol Biol. 2006, 15: 1-12. 10.1111/j.1365-2583.2006.00610.x.CrossRefPubMed
20.
go back to reference Cook PE, Sinkins SP: Transcriptional profiling of Anopheles gambiae mosquitoes for adult age estimation. Insect Mol Biol. 2010, 19: 745-751. 10.1111/j.1365-2583.2010.01034.x.PubMedCentralCrossRefPubMed Cook PE, Sinkins SP: Transcriptional profiling of Anopheles gambiae mosquitoes for adult age estimation. Insect Mol Biol. 2010, 19: 745-751. 10.1111/j.1365-2583.2010.01034.x.PubMedCentralCrossRefPubMed
21.
go back to reference Wang M-H, Marinotti O, James AA, Walker E, Githure J, Yan G: Genome-wide patterns of gene expression during aging in the African malaria vector Anopheles gambiae. PLoS ONE. 2010, 5: e13359-10.1371/journal.pone.0013359.PubMedCentralCrossRefPubMed Wang M-H, Marinotti O, James AA, Walker E, Githure J, Yan G: Genome-wide patterns of gene expression during aging in the African malaria vector Anopheles gambiae. PLoS ONE. 2010, 5: e13359-10.1371/journal.pone.0013359.PubMedCentralCrossRefPubMed
22.
go back to reference Cook P, Hugo L, Iturbe-Ormaetxe I, Williams C, Chenoweth S, Ritchie S, Ryan P, Kay B, Blows M, O'Neill S: Predicting the age of mosquitoes using transcriptional profiles. Nat Protoc. 2007, 2: 2796-2806. 10.1038/nprot.2007.396.CrossRefPubMed Cook P, Hugo L, Iturbe-Ormaetxe I, Williams C, Chenoweth S, Ritchie S, Ryan P, Kay B, Blows M, O'Neill S: Predicting the age of mosquitoes using transcriptional profiles. Nat Protoc. 2007, 2: 2796-2806. 10.1038/nprot.2007.396.CrossRefPubMed
23.
go back to reference Scott J, Brogdon W, Collins F: Identification of single specimens of the Anopheles gambiae c omplex by the polymerase chain reaction. Am J Trop Med Hyg. 1993, 49: 520-529.PubMed Scott J, Brogdon W, Collins F: Identification of single specimens of the Anopheles gambiae c omplex by the polymerase chain reaction. Am J Trop Med Hyg. 1993, 49: 520-529.PubMed
24.
go back to reference Bass C, Williamson M, Wilding C, Donnelly M, Field L: Identification of the main malaria vectors in the Anopheles gambiae species complex using a TaqMan real-time PCR assay. Malar J. 2007, 6: 155-10.1186/1475-2875-6-155.PubMedCentralCrossRefPubMed Bass C, Williamson M, Wilding C, Donnelly M, Field L: Identification of the main malaria vectors in the Anopheles gambiae species complex using a TaqMan real-time PCR assay. Malar J. 2007, 6: 155-10.1186/1475-2875-6-155.PubMedCentralCrossRefPubMed
25.
go back to reference Vezenegho S, Bass C, Puinean M, Williamson M, Field L, Coetzee M, Koekemoer L: Development of multiplex real-time PCR assays for identification of members of the Anopheles funestus species group. Malar J. 2009, 8: 282-10.1186/1475-2875-8-282.PubMedCentralCrossRefPubMed Vezenegho S, Bass C, Puinean M, Williamson M, Field L, Coetzee M, Koekemoer L: Development of multiplex real-time PCR assays for identification of members of the Anopheles funestus species group. Malar J. 2009, 8: 282-10.1186/1475-2875-8-282.PubMedCentralCrossRefPubMed
26.
go back to reference Mayagaya VS, Michel K, Benedict MQ, Killeen GF, Wirtz RA, Ferguson HM, Dowell FE: Non-destructive determination of age and species of Anopheles gambiae s.l. using near-infrared spectroscopy. Am J Trop Med Hyg. 2009, 81: 622-630. 10.4269/ajtmh.2009.09-0192.CrossRefPubMed Mayagaya VS, Michel K, Benedict MQ, Killeen GF, Wirtz RA, Ferguson HM, Dowell FE: Non-destructive determination of age and species of Anopheles gambiae s.l. using near-infrared spectroscopy. Am J Trop Med Hyg. 2009, 81: 622-630. 10.4269/ajtmh.2009.09-0192.CrossRefPubMed
27.
go back to reference Sikulu M, Killeen G, Hugo L, Ryan P, Dowell K, Wirtz R, Moore S, Dowell F: Near-infrared spectroscopy as a complementary age grading and species identification tool for African malaria vectors. Parasites & Vectors. 2010, 3: e49-10.1186/1756-3305-3-49.CrossRef Sikulu M, Killeen G, Hugo L, Ryan P, Dowell K, Wirtz R, Moore S, Dowell F: Near-infrared spectroscopy as a complementary age grading and species identification tool for African malaria vectors. Parasites & Vectors. 2010, 3: e49-10.1186/1756-3305-3-49.CrossRef
28.
go back to reference Gray E, Bradley T: Physiology of dessication resistance in Anopheles gambiae and Anopheles arabiensis. Am J Trop Med Hyg. 2005, 73: 553-559.PubMed Gray E, Bradley T: Physiology of dessication resistance in Anopheles gambiae and Anopheles arabiensis. Am J Trop Med Hyg. 2005, 73: 553-559.PubMed
29.
go back to reference Quicke DLJ, Belshaw R, Lopez-Vaamonde C: Preservation of hymenopteran specimens for subsequent molecular and morphological study. Zoologica Scripta. 1999, 28: 261-267. 10.1046/j.1463-6409.1999.00004.x.CrossRef Quicke DLJ, Belshaw R, Lopez-Vaamonde C: Preservation of hymenopteran specimens for subsequent molecular and morphological study. Zoologica Scripta. 1999, 28: 261-267. 10.1046/j.1463-6409.1999.00004.x.CrossRef
30.
go back to reference Perez-Mendoza J, Dowell FE, Broce AB, Throne JE, Wirtz RA, Xie F, Fabrick JA, Baker JE: Chronological age-grading of house flies by using near-infrared spectroscopy. J Med Ent. 2002, 39: 499-508. 10.1603/0022-2585-39.3.499.CrossRef Perez-Mendoza J, Dowell FE, Broce AB, Throne JE, Wirtz RA, Xie F, Fabrick JA, Baker JE: Chronological age-grading of house flies by using near-infrared spectroscopy. J Med Ent. 2002, 39: 499-508. 10.1603/0022-2585-39.3.499.CrossRef
31.
go back to reference Hugo LE, Cook PE, Johnson PH, Rapley LP, Kay BH, Ryan PA, Ritchie SA, O'Neill SL: Field validation of a transcriptional assay for the prediction of age of uncaged Aedes aegypti mosquitoes in norther Australia. PLoS Neglected Trop Dis. 2010, 4: e608-10.1371/journal.pntd.0000608.CrossRef Hugo LE, Cook PE, Johnson PH, Rapley LP, Kay BH, Ryan PA, Ritchie SA, O'Neill SL: Field validation of a transcriptional assay for the prediction of age of uncaged Aedes aegypti mosquitoes in norther Australia. PLoS Neglected Trop Dis. 2010, 4: e608-10.1371/journal.pntd.0000608.CrossRef
32.
go back to reference Gorokhova E: Effects of preservation an dstorage of microcrustaceans in RNAlater on RNA and DNA degradation. Limnol Oceanogr: Methods. 2005, 3: 143-148.CrossRef Gorokhova E: Effects of preservation an dstorage of microcrustaceans in RNAlater on RNA and DNA degradation. Limnol Oceanogr: Methods. 2005, 3: 143-148.CrossRef
33.
go back to reference Ferguson H, Ng'habi K, Walder T, Kadungula D, Moore S, Lyimo I, Russell T, Urassa H, Mshinda H, Killeen G, Knols B: Establishment of a large semi-field system for experimental study of African malaria vector ecology and control in Tanzania. Malar J. 2008, 7: e158-10.1186/1475-2875-7-158.CrossRef Ferguson H, Ng'habi K, Walder T, Kadungula D, Moore S, Lyimo I, Russell T, Urassa H, Mshinda H, Killeen G, Knols B: Establishment of a large semi-field system for experimental study of African malaria vector ecology and control in Tanzania. Malar J. 2008, 7: e158-10.1186/1475-2875-7-158.CrossRef
34.
go back to reference Perez-Mendoza J, Throne JE, Dowell FE, Baker JE: Chronological age-grading of three species of stored- product beetles by using near-infrared spectroscopy. J Econ Ent. 2002, 97: 1159-1167.CrossRef Perez-Mendoza J, Throne JE, Dowell FE, Baker JE: Chronological age-grading of three species of stored- product beetles by using near-infrared spectroscopy. J Econ Ent. 2002, 97: 1159-1167.CrossRef
35.
go back to reference Reeves WK, Peiris KHS, Scholte EJ, Wirtz RA, Dowell FE: Age-grading the biting midge Culicoides sonorensis using near-infrared spectroscopy. Med Vet Ent. 2010, 24: 32-37. 10.1111/j.1365-2915.2009.00843.x.CrossRef Reeves WK, Peiris KHS, Scholte EJ, Wirtz RA, Dowell FE: Age-grading the biting midge Culicoides sonorensis using near-infrared spectroscopy. Med Vet Ent. 2010, 24: 32-37. 10.1111/j.1365-2915.2009.00843.x.CrossRef
36.
go back to reference Aldrich BT, Maghirang EB, Dowell FE, Kambhampati S: Identification of termite species and subspecies of the genus Zootermopsis using near-infrared reflectance spectroscopy. J Insect Sci. 2007, 7: 1-7.CrossRef Aldrich BT, Maghirang EB, Dowell FE, Kambhampati S: Identification of termite species and subspecies of the genus Zootermopsis using near-infrared reflectance spectroscopy. J Insect Sci. 2007, 7: 1-7.CrossRef
Metadata
Title
Evaluating RNAlater® as a preservative for using near-infrared spectroscopy to predict Anopheles gambiae age and species
Authors
Maggy Sikulu
Kayla M Dowell
Leon E Hugo
Robert A Wirtz
Kristin Michel
Kamaranga HS Peiris
Sarah Moore
Gerry F Killeen
Floyd E Dowell
Publication date
01-12-2011
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2011
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/1475-2875-10-186

Other articles of this Issue 1/2011

Malaria Journal 1/2011 Go to the issue
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.