|SHORT RESEARCH COMMUNICATION
|Year : 2018 | Volume
| Issue : 3 | Page : 235-238
Adaptation of Aedes aegypti to salinity: Characterized by larger anal papillae in larvae
SN Surendran1, K Sivabalakrishnan1, T.T.P. Jayadas1, S Santhirasegaram1, A Laheetharan2, M Senthilnanthanan3, R Ramasamy4
1 Department of Zoology, Faculty of Science, University of Jaffna, Jaffna, Sri Lanka
2 Department of Mathematic and Statistics, Faculty of Science, University of Jaffna, Jaffna, Sri Lanka
3 Department of Chemistry, Faculty of Science, University of Jaffna, Jaffna, Sri Lanka
4 ID-FISH Technology Inc., Palo Alto, USA
|Date of Submission||18-Oct-2017|
|Date of Acceptance||25-Jul-2018|
|Date of Web Publication||4-Jan-2019|
S N Surendran
Department of Zoology, University of Jaffna, Jaffna–40000
Source of Support: None, Conflict of Interest: None
Keywords: Aedes aegypti; anal papillae; brackish water; fresh water; salinity-tolerant mosquitoes; Sri Lanka; vector biology
|How to cite this article:|
Surendran S N, Sivabalakrishnan K, Jayadas T, Santhirasegaram S, Laheetharan A, Senthilnanthanan M, Ramasamy R. Adaptation of Aedes aegypti to salinity: Characterized by larger anal papillae in larvae. J Vector Borne Dis 2018;55:235-8
|How to cite this URL:|
Surendran S N, Sivabalakrishnan K, Jayadas T, Santhirasegaram S, Laheetharan A, Senthilnanthanan M, Ramasamy R. Adaptation of Aedes aegypti to salinity: Characterized by larger anal papillae in larvae. J Vector Borne Dis [serial online] 2018 [cited 2019 Nov 22];55:235-8. Available from: http://www.jvbd.org/text.asp?2018/55/3/235/249482
Aedes aegypti is considered the principal vector of several human arboviral diseases, including dengue, chikungunya, West-Nile encephalitis and Zika. Many of the approximately 5% of mosquito species that are salinity tolerant, are vectors of important human and animal diseases; and such vectors can increase in number and invade to new territories in the context of global warming and other anthropogenic environmental changes,,. Aedes aegypti has previously been regarded to oviposit and undergo preimaginal development only in fresh water (FW) collections, and hence, larval control efforts are solely directed towards FW habitats in the vicinity of human dwellings,. We recently showed that Ae. aegypti and the closely related arboviral vector Ae. albopictus can undergo preimaginal development in brackish water (within discarded food and beverage containers) of up to 15 parts per thousand (ppt) salt and 14 ppt salt, respectively in northern Sri Lankan beaches,. Saline, brackish water (BW) and FW are defined as containing >30, 0.5–30 and <0.5 ppt salt, respectively in this context. Aedes aegypti larvae have also been observed in brackish domestic wells of up to 9 ppt salt in the coastal Jaffna peninsula of northern Sri Lanka. Similarly, BW development of Ae. aegypti or Ae. albopictus has been observed in coastal locations of Brunei Darussalam, USA and Brazil. Physiological changes accompanying BW adaptation in Ae. aegypti include the greater ability to oviposit in 10 ppt salinity; and significantly greater, and partly inheritable, larval salinity tolerance.
The anal papillae of Ae. aegypti are reported to transport Na+ and Cl– from a FW environment into the hemolymph,. There is also a salinity-related differential expression of specific aquaporins in Ae. aegypti anal papillae. These findings suggest a possible role of anal papillae in short-term osmoregulation in Ae. aegypti larvae. In an attempt to explore the mechanisms, underlying the adaptation to BW, this study investigated the changes in anal papillae size in Ae. aegypti mosquitoes adapting to BW.
Self-mating laboratory colonies of Ae. aegypti were established from larvae collected from FW ovitraps in Thirunelvely (9° 41' N: 80° 1' E) and BW domestic wells in coastal Kurunagar (9° 39' N: 80° 1' E) in the Jaffna peninsula of Sri Lanka. The FW colony was maintained in potable tap water supplied from a dug-well located in the University of Jaffna campus in Thirunelvely, and the BW colony in sea water collected from Jaffna lagoon, diluted with tap water to yield ~10 ppt salt (measured by a refractor Salinometer, Atago, Japan). The adult and larvae were maintained at 28–30°C with relative humidity ~75% and 12 h dark and light conditions. Eggs from the BW and FW colonies were collected in 10 ppt saline BW and 0 ppt saline FW egg-laying surfaces, respectively and allowed to hatch into larvae in the corresponding BW and FW. Larvae were reared in 24 × 16 cm plastic trays with 1.5 litre water with maximum number of 150 larvae per tray and fed with fish meal powder thrice a day. Adult females were blood-fed on mice according to a protocol approved by the Institutional Animal Ethics Review Committee of the University of Jaffna (June, 2014; AERC/2014/02).
The FW colony maintained up to 20 generations in the laboratory and BW colony maintained up to 24 generations were used for experimental purposes. For experiments to determine the effects of reversing larval rearing salinity (salinity reversal experiments), FW colony eggs were collected on FW egg-laying surfaces and then reared in 10 ppt salinity BW, and also separately in FW in parallel. For the corresponding reversal of salinity, BW colony eggs were collected on BW egg-laying surfaces and reared in FW, and also parallelly in 10 ppt salinity BW.
On an average, >90% of the larvae in the rearing trays in each experiment were of the same age and size. Late III instar larvae from the four test samples were identified as such by counting the number of comb spines and pecten teeth. For measuring the size of anal papillae, late III instar larvae were dorso-ventrally placed individually on a clean glass microscope slide and the four lobes were individually termed I–IV as shown in [Figure 1]a. The maximal length and width of individual anal papillae were measured as indicated in [Figure 1]b using an CX21 (Olympus, Tokyo) light microscope equipped with an ocular micrometer. The maximum lengths of the larvae were also measured at the same time. All the measurements were done and recorded independently by two persons and the average value was used for statistical analysis.
|Figure 1: (a) Photograph of posterior portion of dorso-ventrally placed larvae showing the numbering system of anal papilla lobes; and (b) Photograph of anal papillae of (i) FW larvae, and (ii) BW larvae.|
Click here to view
The Student's two-tailed t-test was performed when mean dimensions of specific lobes were initially compared. Tukey's method for multiple comparisons in the analysis of variance (ANOVA) was used to compare group means at 95% CI, where dimensions of all four anal papillae lobes were compared in salinity reversal experiments.
Delayed pre-imaginal development previously observed during the early stages of BW adaptation (up to five generations) in Ae. aegypti was not observed in the present experiments, most likely due to the longer time of adaptation to salinity. Initially, the length of a single anal papilla was measured. The mean anal papilla IV length of BW larvae (15th generation) was 0.55 ± 0.5 mm, and this was significantly greater than the mean anal papilla IV length of FW (10th generation) larvae of 0.50 ± 0. 04 mm (p < 0.05). In a subsequent experiment, the mean anal papilla IV length of BW (18th generation) larvae was 0.57± 0.07 mm and this was significantly greater than the mean anal papilla IV length of FW (14th generation) larvae of 0. 45 ± 0.06 mm.
As the initial measurements suggested that there were significant differences in the sizes of anal papillae between FW and BW larvae, the experiment was further modified to measure the length and width of the four anal papillae (I–IV) that involved reversing larval-rearing salinity. The maximum length (L-I to L-IV) and corresponding width (W-I to W-IV) of each anal papilla and the total length of the larvae [Figure 1]a were measured in 10 late III instar larvae of FW (generation 20) and BW (generation 24). The dimensions of the late III larvae anal papillary lobes and the outcome of the statistical analysis are presented in [Table 1]. The BW larvae reared in BW possessed significantly larger anal papillae lobes in both length and width than FW larvae reared in FW [Figure 1]b, [Table 1]. When FW larvae were shifted to 10 ppt salinity BW for rearing, they tended to have longer and wider lobes than FW larvae maintained in FW, but the differences were not statistically significant. However, when the larvae from the BW colony larvae were subjected to a reversal in salinity and reared in FW, there was a significant reduction in their mean lobe length and width. The BW colony larvae reared in FW had anal papillae that were significantly longer (but not significantly wider) than FW colony larvae that were reversed to develop in BW [Table 1]. Although larvae developing in BW tended to have longer mean whole larval lengths than larvae developing in FW, statistically significant differences could not be established except that BW colony larvae in BW were longer than in BW colony eggs allowed to develop as larvae in FW [Table 1].
|Table 1: Morphometric analysis of larval anal papillae of Ae. aegypti reared in brackish and fresh water|
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The results, therefore indicate that the anal papillae of Ae. aegypti larvae increase in size when adapted to BW and this is partly reversed when they are reared in FW. The corresponding changes in larvae of Ae. aegypti adapted to FW and reared in BW, although not statistically significant, tend to support this conclusion.
The only other study to our knowledge on size variations in anal papillae of Ae. aegypti in response to salinity changes was performed in UK in 1938. This reported that when larvae of long established laboratory colonies of Ae. aegypti maintained in FW are placed in solutions of increasing NaCl concentration up to 0.9% or 9 ppt, the length of the anal papillae decreased up to 17%. The changes were attributed to atrophy of the anal papillae associated with a decreased need to take up NaCl to maintain hemolymph NaCl concentrations in a more saline environment. The different findings of this study might be due to the use of larvae from colonies adapted relatively to FW and BW from recently field collections made in the Jaffna peninsula rather than a long established FW Ae. aegypti colony in UK, and the use of diluted sea water in present experiments instead of NaCl solutions to better reflect the natural coastal BW habitats of Ae. aegypti.
Larvae of some salinity-tolerant mosquito species have evolved various physiological mechanisms associated with specific anatomical changes to cope with salinity in the larval environment. Aedes taeniorhynchus ingest the surrounding fluid and excrete Na+ and Cl– from an additional differentiated rectal segment (the posterior rectum) to produce a hyperosmotic urine. Larvae of the euryhaline malaria vector An. albimanus are able to differentially localize sodium-potassium ATPase in specialized anterior dorsal cells of their rectum in fresh or saline water for likely osmoregulation through ion excretion. There is no evidence yet to suggest that Ae. aegypti have specialized rectal structures that are associated with salinity adaptation. The present findings, and related physiological observations on anal papillae,,, lead us to propose that changes in ion and water transport in Ae. aegypti anal papillae are reflected in size change, and that the underlying osmoregulatory mechanisms may be critical to permit their preimaginal development in BW habitats.
In conclusion, the potential importance of BW adaptation in Ae. aegypti to their anthropogenically-induced spread and increased transmission of arboviral diseases,, warrant further studies to elucidate the relevant biological adaptive mechanisms, e.g. through ongoing differential gene expression studies on anal papillae of Ae. aegypti. This is important since the BW developing Ae. aegypti and Ae. albopictus originating in the Jaffna peninsula, like their FW counterparts, can be infected with dengue virus and transmit the virus transovarially to their progeny.
| References|| |
Bradley TJ. Physiology of osmoregulation in mosquitoes. Annu Rev Entomol
Ramasamy R, Surendran SN. Possible impact of rising sea levels on vector-borne infectious diseases. BMC Infect Dis
Ramasamy R, Surendran SN. Mosquito vectors developing in atypical anthropogenic habitats—global overview of recent observations, mechanisms and impact on disease transmission. J Vector Borne Dis
Ramasamy R. Adaptation of fresh water mosquito vectors to salinity increases arboviral disease transmission risk in the context of anthropogenic environmental changes. In: Shapshak P, Sinnott JT, Somboonwit C, Kuhn JH, editors. Global Virology I– Identifying and Investigating Viral Diseases
. New York, Springer 2015; p. 45–54.
Barraud PJ. The fauna of British India, including Ceylon and Burma. Diptera, Vol V. Family Culicidae: Tribes Megarhinini and Culicini. London, UK: Taylor and Francis 1934; p. 463.
Ramasamy R, Surendran SN, Jude PJ, Dharshini S, Vinobaba M. Larval development of Aedes aegypti
and Aedes albopictus
in peri-urban brackish water and its implications for transmission of arboviral diseases. Plos Negl Trop Dis
2011; 5(11): e1369.
Jude PJ, Tharmasegaram T, Sivasubramaniyam G, Senthilnanthnan M, Kannathasan S, Raveendran S, et al
. Salinity-tolerant larvae of mosquito vectors in the tropical coast of Jaffna, Sri Lanka and the effect of salinity on the toxicity of Bacillus thuringiensis
to Aedes aegypti
larvae. Parasit Vectors
Surendran SN, Jude PJ, Thabothiny V, Raveendran S, Ramasamy R. Preimaginal development of Aedes aegypti
in brackish and fresh water urban domestic wells in Sri Lanka. J Vector Ecol
Idris FH, Usman A, Surendran SN, Ramasamy R. Detection of Aedes albopictus
pre-imaginal stages in brackish water habitats in Brunei Darussalam. J Vector Ecol
2013; 38(1): 197–9.
Yee DA, Himel E, Reiskind MH, Vamosi SM. Implications of saline concentrations for the performance and competitive interactions of the mosquitoes Aedes aegypti (Stegomyia aegypti)
and Aedes albopictus (Stegomyia albopictus). Med Vet Entomol
2014; 28(1): 60–9.
Arduino MB, Mucci LF, Serpa LLN, Rodrigues MM. Effect of salinity on the behaviour of Aedes aegypti
populations from the coast and plateau of southeastern Brazil. J Vector Borne Dis
2015; 52(1): 79–87.
Ramasamy R, Jude PJ, Veluppillai T, Eswaramohan T, Surendran SN. Biological differences between brackish and fresh water-derived Aedes aegypti
from two locations in the Jaffna Peninsula of Sri Lanka and the implications for arboviral disease transmission. PLoS One
Donini A, O'Donnell MJ. Analysis of Na+
concentration gradients adjacent to the surface of anal papillae of the mosquito Aedes aegypti:
Application of self-referencing ion-selective microelectrodes. J Exp Biol
Donini A, Gaidhu MP, Strasberg DR, O'Donnell MJ. Changing salinity induces alterations in hemolymph ion concentrations and Na+
and Cl- transport kinetics of the anal papillae in the larval mosquito, Aedes aegypti. J Exp Biol
Akhter H, Misyura L, Bui P, Donini A. Salinity responsive aquaporins in the anal papillae of the larval mosquito Aedes aegypti. Comp Biochem Physiol A Mol Integr Physiol
Christophers SR. Aedes aegypti
(L.), the yellow fever mosquito: Its life history, bionomics and structure. New York: Cambridge University Press 1960; p. 54–8.
Andrew J, Bar A. Morphology and morphometry of Aedes aegypti
larvae. Ann Rev Res Biol
2013; 3(1): 1–21.
Wigglesworth VB. The regulation of osmotic pressure and chloride concentration in the haemolymph of mosquito larvae. J Exp Biol
Smith KE, VanEkeris LA, Okech BA, Harvey WH, Linser PJ. Larval anopheline mosquito recta exhibit a dramatic change in localization patterns of ion transport proteins in response to shifting salinity: A comparison between anopheline and culicine larvae. J Exp Biol
(Pt 19): 3067–76.
Surendran SN, Veluppillai T, Eswaramohan T, Sivabalakrishnan K, Noordeen F, Ramasamy R. Salinity tolerant Aedes aegypti and Aedes albopictus—Infection with dengue virus and contribution to dengue transmission in a coastal peninsula. J Vector Borne Dis 2018; 55(1): 26–33.