Biol Invasions

DOI 10.1007/s10530-010-9721-z

1 ORIGINAL PAPER

2 Aedes aegypti and Aedes albopictus in Bermuda: extinction,

3 invasion, invasion and extinction
4 Laran Kaplan • David Kendell •
Author Proof

5 Deborah Robertson • Todd Livdahl •

6 Camilo Khatchikian

7 Received: 3 July 2009 / Accepted: 8 February 2010

8 Ó Springer Science+Business Media B.V. 2010

9 Abstract We provide an analysis of the invasion the arrival of A. albopictus in the year 2000. Possible 27
10 and spread of the container inhabiting mosquitoes mechanisms for this rapid decline at relatively low 28
11 Aedes aegypti and Aedes albopictus in the Bermuda density of the second invader are discussed in the 29
12 Islands. Considered eradicated in the mid-1960s, context of classical competition theory and earlier 30
13 A. aegypti was redetected in 1997, and A. albopictus experimental results from Florida, as well as alterna- 31
14 was first detected in 2000. Based on weekly ovitrap tive explanations. We suggest that support for the 32
15 data collected during the early stages of the invasion, competition hypothesis to account for the decline of 33
16 we mapped the spread of Aedes throughout the A. aegypti is ambiguous and likely to be an incom- 34
17 islands. We analyzed the effects of buildings and plete explanation. 35
18 roads on mosquito density and found a significant
19 association between density and distance to roads, but Keywords Aedes albopictus  Aedes aegypti  36
20 not to buildings. We discuss the potential role of Vector invasion  Bermuda Islands  37
21 human transport in the rapid spread in the islands. Competition 38
40
22 The temporal correlation in ovitrap collection values
39
23 decreased progressively, suggesting that habitat deg-
24 radation due to control efforts were responsible for
25 local shifts in mosquito densities. We report a sharp Introduction 41
26 decrease in A. aegypti presence and abundance after
Aedes albopictus and Aedes aegypti are mosquitoes 42
(Diptera: Culicidae) that inhabit small containers as 43
A1 L. Kaplan  D. Robertson  T. Livdahl (&) 
larvae, invasive to the Bermuda Islands (UK). 44
A2 C. Khatchikian
A3 Department of Biology, Clark University, Worcester, A. albopictus originates from southeast Asia but has 45
A4 MA 01610, USA dramatically expanded its range to many tropical, 46
A5 e-mail: tlivdahl@clarku.edu subtropical and temperate regions, both in urban and 47
rural areas around the world (e.g., Benedict et al. 48
A6 D. Kendell
A7 Bermuda Ministry of Health, Point Finger Road, 2007; Hawley 1988; Lounibos 2002; Novak 1992; 49
A8 Bermuda, UK Urbanelli et al. 2000). A. aegypti originates from 50
Africa and spread to many areas in the tropics, 51
A9 Present Address:
especially in concurrence with the slave trade 52
A10 L. Kaplan
A11 Center for Vector Biology, Rutgers University, increase in the 16th and 17th century (Lounibos 53
A12 New Brunswick, NJ 08901, USA 2002); currently, both species co-occur in many 54

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55 regions of the world (e.g., Braks et al. 2003; has been suggested as more efficient by Rawlins et al. 104
56 Fontenille and Rodhain 1989; Tabachnick 1991). (1998) and allowed egg collections to be sent out for 105
57 These two mosquito species have prompted consid- analysis and species identification. This monitoring 106
58 erable interest in multiple research areas, including methodology can be somewhat limited, as it only 107
59 vector competence (e.g., Boromisa et al. 1987; Diallo detects the presence of mosquitoes within close 108
60 et al. 2008; Moore et al. 2007), insecticide resistance vicinity, but it could be used as a proxy estimator 109
61 (e.g., Hidayati et al. 2005; Stasiak et al. 1969; of presence and size of mosquito populations in the 110
62 Wesson 1990), spatial, temporal and geographical trap’s vicinity. The Bermuda Islands provide an 111
63 analyses (e.g., Benedict et al. 2007; Castro Gomes interesting opportunity to study the nearly coinciden- 112
Author Proof

64 et al. 2005; Francy et al. 1990; Kobayashi et al. tal invasions of these two ecologically similar 113
65 2002), and ecological and evolutionary studies (e.g., species, taking into consideration the islands’ isola- 114
66 Juliano et al. 2002; Pumpuni et al. 1992). Both tion, small size, and the extensive ovitrap sampling 115
67 species have been involved in historic and current program that allows tracking the mosquitoes’ inva- 116
68 transmission of dengue and yellow fever throughout sion in the islands. 117
69 their range. A. albopictus is a laboratory vector of Despite ecological similarities of A. aegypti and 118
70 more than 30 viruses, although only a few of these A. albopictus in breeding habitat and host selection, 119
71 affect humans, including dengue, Chikungunya, St. Hornby et al. (1994) suggested that they occupy 120
72 Louis, and La Cross encephalitis viruses (Mitchell different niches. In Florida (USA), Brazil, and many 121
73 1995). A. aegypti is the historical vector for yellow regions of Asia, A. albopictus generally prefers rural 122
74 fever virus (e.g., Beaty and Aitken 1979) and has and suburban, underdeveloped or lightly developed 123
75 been reported as the most competent vector of the areas, whereas A. aegypti prefers urban and suburban 124
76 dengue virus (e.g., Gubler 1998). highly developed areas (Braks et al. 2003; Hornby 125
77 The Bermuda Islands (UK), located in the Atlantic et al. 1994; Gilotra et al. 1967; O’Meara et al. 1992, 126
78 Ocean off the east coast of the USA (32°180 N 1995). A. aegypti, which had been found throughout 127
79 64°470 W), have a subtropical climate, with mild Florida for many years prior to the arrival of A. 128
80 winters and hot, humid summers, providing suitable albopictus in the early 1990s, has since declined in 129
81 conditions for both species of mosquitoes. Histori- abundance and distribution (Hornby et al. 1994; 130
82 cally, A. aegypti was responsible for extensive O’Meara et al. 1995). This decline in A. aegypti’s 131
83 outbreaks of yellow fever and dengue fever during abundance occurred soon after the establishment of 132
84 the past few centuries. In the 1940s, a large A. albopictus in locations such as scrap tire sites and 133
85 control campaign was undertaken and A. aegypti rural areas where the species’ distributions over- 134
86 was considered eradicated by the 1960s (Camargo lapped (O’Meara et al. 1995). Lounibos (2002) 135
87 1967). However, A. aegypti was rediscovered in 1997 suggests that the decline has been due to a combi- 136
88 and subsequently, in 2000, A. albopictus was also nation of several factors, including A. albopictus 137
89 found in the islands by health officers. Before the superiority in larval resource competition. In the 138
90 rediscovery of Aedes in the islands, the Health United States, a correlation was found between the 139
91 Department of Bermuda maintained a mosquito- introduction and increase of A. albopictus and a more 140
92 monitoring program only at ports of entry, but after restricted distribution and declining abundance in A. 141
93 the rediscovery of A. aegypti, the program was aegypti (O’Meara et al. 1995). 142
94 expanded. Ovitraps were installed to monitor mos- Juliano (1998) examined competition between the 143
95 quito prevalence and spread across the islands, two species. A. albopictus was the dominant com- 144
96 comprising, by 2005, 582 weekly collected ovitraps petitor in experimental field containers, with positive 145
97 distributed throughout the limited Bermuda surface population growth in treatments with low resource 146
98 (\54 km2). The ovitrap program is paired with a availability and treatments with a high combined 147
99 permanent, media and field based resident conscious- species density. Overall, the experiments pointed to 148
100 ness program in conjunction with crews deployed in A. albopictus as the superior competitor, considering 149
101 the ground cleaning and surveying containers and its higher rate of survival to adulthood. A. aegypti 150
102 other breeding areas. Monitoring mosquito popula- was only able to complete its development in 151
103 tions by ovitrap collections rather than larval surveys treatments with the lowest density or in treatments 152

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153 with leaf litter added to the filtered tire water medium Willey 1991). The critical parameters to estimate 202
154 (Juliano 1998). A similar experiment performed in for quantitative prediction of this interaction are 203
155 Brazil also supports A. albopictus’ competitive competition coefficients (a) and carrying capacities 204
156 advantage under field situations. A. albopictus was (K). Although a number of studies of A. albopictus 205
157 suggested as the dominant larval competitor consid- and A. aegypti have been conducted yielding data that 206
158 ering that populations maintained positive growth could have provided them, none of the studies of A. 207
159 rates in the field at higher combined densities and albopictus and A. aegypti have estimated these 208
160 lower per capita resource availability than A. aegypti parameters. 209
161 (Braks et al. 2004). Several studies have reported that Considering the habitat-specificity of these param- 210
Author Proof

162 detritus may affect competitive outcome, with vary- eters, it is remarkably difficult to obtain estimates of 211
163 ing results; A. albopictus outcompetes A. aegypti in carrying capacity or competition coefficients in non- 212
164 many detritus types, but some food sources, such as experimental field conditions. Nevertheless, it seems 213
165 liver powder, may favor A. aegypti (Black et al. likely that those parameters would be similar in 214
166 1989), or allow stable coexistence of the two species ecologically similar species. If this is the situation, 215
167 (Murrell and Juliano 2008). the isoclines of the two species would be close 216
168 The coincident decline in several A. aegypti together, as shown in Fig. 1a, and the inferior 217
169 populations that occurred after A. albopictus inva- competitor would begin to decline only when the 218
170 sions may have resulted from competitive displace- better competitor is near its equilibrium density. 219
171 ment by exploitation, considering the apparent A. Exclusion should be a slow process in such a case 220
172 albopictus superiority in field and laboratory condi- (Fig. 1b). A much more rapid exclusion is possible if 221
173 tions (e.g., Braks et al. 2004; Juliano 1998). Never- carrying capacities differ markedly, or if competition 222
174 theless, alternative scenarios are possible, including coefficients yield a dramatic asymmetry in the impact 223
175 interference between species or some other types of of one species on the other. An illustration of two 224
176 interactions (reviewed by Lounibos 2002, 2007) that species with quite different carrying capacities 225
177 could result in similar outcomes: mating interference, appears in Fig. 1c, d. The A. albopictus invasion of 226
178 microparasite infections, and hatching inhibition. In the Bermuda Islands, previously occupied by 227
179 addition, it has been reported that predatory species A. aegypti, provides a novel opportunity to examine 228
180 such as Corethrella appendiculata or Toxorhynchites this possibility, taking advantage of the extensive 229
181 rutilus could change species interaction outcome in ovitrap program, the small area of Bermuda, and the 230
182 small containers in the US (e.g., Griswold and fairly homogeneous, urbanized environment. 231
183 Lounibos 2006; Juliano et al. 2009; Kesavaraju We report here the invasion, progress, spread, 232
184 et al. 2008). However, these predator species are and the contrasting outcomes of A. aegypti and 233
185 not present in Bermuda, and there is no other predator A. albopictus invasions in the Bermudas Islands. In 234
186 occupying small containers in the islands (D. Ken- addition, we aim to evaluate the A. albopictus 235
187 dell, pers. obs.). competitive superiority hypothesis by examining 236
188 Various approaches have been taken to model indirect indicators of competitive exclusion. 237
189 competition, most of which trace to the Lotka-
190 Volterra model, which is among the simplest descrip-
191 tions of two species sharing some common resource
192 base. The combination of conditions necessary for Methods 238
193 competitive superiority to explain the displacement
194 of one species by another by the Lotka-Volterra Sampling 239
195 model are depicted in Fig. 1. The species whose
196 isocline is higher is able to increase in number, Ovitraps consisted of wide-mouthed amber glass jars 240
197 driving the other species down to extinction when (15.2 9 7.6 cm), half-filled with rain water, with a 241
198 reaching its carrying capacity. This analysis has been masonite paddle (2 9 16 cm) inserted as an ovipo- 242
199 used in similar circumstances to model the interaction sition substrate. All ovitraps in the sampling program 243
200 between A. albopictus and a North American native were monitored weekly by the Bermuda Ministry of 244
201 container inhabiting, A. triseriatus (Livdahl and Health, starting with 135 ovitraps by 2000, expanded 245

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Fig. 1 General representation of the Lotka-Volterra competi-
tion model for competitive exclusion, represented in a and c
with a phase diagram of population size of species 1 (N1,
x-axis) versus population size of species 2 (N2, y-axis). Zero
growth isocline for species 2 is located above the correspond-
ing isocline of species 1; species 2 is a superior competitor and
eventually drives species 1 to extinction when reaching its
carrying capacity. Specific location of isoclines depend on
carrying capacities for both species (K1, K2) and the relative
competitive coefficients (a12, a21). Arrows depict general
246 to 300 ovitraps by 2003, and expanded again in
247 2005–582 ovitraps.
248 Species identification
249 To identify the mosquito species present in the
250 samples, masonite paddles containing eggs from the
251 ovitraps collected in Bermuda from 2003 to 2005
252 were sent to Clark University. All paddles were
253 sorted by collection site and year. Using a toothpick
254 wet with tap water, eggs were removed from paddles
255 and placed into 0.6 ml tubes. Each individual paddle
256 constituted one sample, and 3–15 eggs were used per
257 sample. If more than 15 eggs were found on a paddle,
258 a random sample was taken. For this study, six to
259 eight samples were taken from each site (n = 39) on
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directions of change within each region, and a trajectory is
shown with a dashed line. b, d Illustrate the densities of each
species through time for the trajectories shown in the phase
diagrams on the left. For clarity, a12, a21 = 1 in both
simulations, but K1 = 90 in the top simulation and 50 in the
bottom case, with K2 = 100 in both cases. Note that the
situation in which the species have the most different carrying
capacities results in a much more rapid displacement, at a
much lower density of the dominant species
the island for each year. DNA was extracted using the 260
E.Z.N.A. Forensic DNA Kit (Omega Bio-tek) in 261
350 ll of STL buffer solution and eggs were 262
disrupted by mechanical agitation for 4 min in the 263
presence of 3.2 mm chrome-steel balls (Mini Bead- 264
beater 8, BioSpec Products, Inc.). Genomic DNA was 265
extracted following the manufacturer’s recommenda- 266
tions, except that HB buffer was not used and DNA 267
was eluted in 55 ll of elution buffer. 268
The ITS2 region of each species was amplified 269
using 28S (50 -TCACACATTATTTGAGGCCTAC- 270
30 ) and 5.8S (50 -TGTGAACTGCAGGACACATG- 271
30 ) primers previously shown to produce different 272
sized products from A. aegypti and A. albopictus 273
(Wesson et al. 1992). PCR reactions (50 ll) consisted 274
of 15 ll of the extracted DNA solution, 2.5 ll of each 275
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276 primer (10 lM stock concentration), 25 ll Promega assess the temporal correlation in number of eggs per 323
277 PCR Master Mix and 5 ll of sterile, distilled water. ovitrap per year, a modified t-test for correlation was 324
278 After an initial denaturing step at 94°C for 3.5 min, performed using the Clifford, Richardson, and 325
279 the following cycling parameters (30 cycles) were Hémon (CRH) procedure, which corrects for spatial 326
280 used: 94°C for 1 min, 50°C for 1 min, and 72°C for autocorrelation. The CRH test was implemented in 327
281 1 min with a final extension at 72°C for 5 min. PCR Passage v.1.0 (Rosenberg 2001), using scaled euclid- 328
282 products were analyzed on a 1% agarose gel using ian distances and 50 equal observation classes. 329
283 standard electrophoretic procedures. DNA from pre- Significant correlation values after Dutilleul’s cor- 330
284 viously identified eggs from both species was used as rection were regressed against time interval. 331
Author Proof

285 standards to verify that PCR products from both Taking into consideration the progressive increase 332
286 species were obtained from mixed-species samples. in the weekly number of ovitraps deployed in the 333
287 Samples were scored as positive for a given species if islands, we chose two statistics to estimate habitat use 334
288 an appropriately-sized PCR product (A. aegypti and crowding levels, and to describe the trajectory of 335
289 324 bp; A. albopictus 518 bp) was visible on the Aedes populations in the islands: the average number 336
290 agarose gels. of eggs per ovitrap, and the proportion of positive 337
ovitraps. No satisfactory transformation for normality 338
291 Analysis was found for these statistics (untransformed average 339
number of eggs W = 0.67, untransformed proportion 340
292 To map the Aedes spread across the island, contin- of positive ovitraps W = 0.81, both P \ 0.001), so 341
293 uous surfaces were interpolated from the yearly totals the data were analyzed using non-parametric 342
294 for each ovitrap using Idrisi v.15.0 Los Andes Edition ANOVA. For pairwise comparisons, bootstrap pro- 343
295 (Clark Labs 2007) INTERPOL module with a cedures were utilized to produce 95% confidence 344
296 distance weight of 2.0 and six points search radio intervals after 1,000 replicates using S-plus v.8.0.4 345
297 option. The values obtained using this procedure (Insightful Corp 2007). Nonoverlapping confidence 346
298 allow a relative estimation of oviposition activity in intervals were considered statistically different. 347
299 the ovitrap’s vicinity. Due to the low number of An ordinal logistic regression (R2 = 0.45, P \ 348
300 ovitraps on the ground during the years 2000–2002, 0.05, lack of fit P = 0.64) was used to estimate A. 349
301 caution should be taken when reading those surfaces aegypti decline based on PCR detection for the years 350
302 as the interpolation algorithm is likely to produce 2003–2005 and the known presence of the species as 351
303 artifacts. All spatial analyses were conducted using the only container inhabiting mosquito in 1999. 352
304 the same 300 georeferenced ovitraps present during Unless stated otherwise, all statistical analyses were 353
305 the years 2003–2004; for years 2000–2002 those performed using JMP 7.0 (SAS Institute Inc. 2007). 354
306 ovitraps were considered missing data.
307 Layers containing the georeferenced location of
308 buildings and roads were used to produce continuous Results 355
309 surfaces with distances to features using Idrisi’s
310 DISTANCE module. The resulting raster layers were Spatial patterns 356
311 imported into ArcMap 9.2 (ESRI 2006) and demo-
312 graphic variable values for the ovitraps locations The spread of unidentified Aedes in the Bermuda 357
313 were extracted using the intersect point tool from Islands was extremely fast. Aedes were detected on 358
314 Hawth’s analysis tool (Beyer 2004). The resulting all populated islands as early as 2003, only 5 years 359
315 distance values failed the Shapiro–Wilk test for after the rediscovery of Aedes aegypti in 1998 and 360
316 normality (untransformed distance to buildings W = 3 years after the first detection of Aedes albopictus in 361
317 0.76, untransformed distance to roads W = 0.49, both 2000 (Fig. 2). A consistent pattern of high egg counts 362
318 P \ 0.001) and several attempted transformations in collections in the southern area was observed 363
319 failed. The distance values were categorized in across the different years, even though the specific 364
320 quartiles and the effect of variables over the total locations and relative values vary to some degree. In 365
321 number of eggs for years 2003–2007 was evaluated addition, ovitraps located near roads, in the first 366
322 with a Van der Waerden non-parametric ANOVA. To distance quartile (from 0 to 4.47 m), had higher egg 367

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 L. Kaplan et al.

were found in traps during every week for at least one 381
of the years of the study period, and larvae can be 382
found in containers throughout the year (D. Kendell, 383
pers. obs.). 384
The temporal correlation in number of eggs per 385
ovitrap per year reported 22 significant (over 28 386
possible combinations) correlations between years 387
considering all possible year combinations. The 388
correlation analysis shows a significant decline in 389
Author Proof

the correlation as the time interval increases 390

(R2 = 0.5, P [ 0.001, Fig. 4). 391

Community composition 392

PCR-based species identification revealed an abrupt 393

decline in A. aegypti populations on Bermuda egg 394
samples collected in ovitraps during 2003–2005. The 395
percentage of samples positive for A. aegypti 396
decreased from 4.35 and 9.09% in 2003 and 2004, 397
respectively, to 3.06% by 2005. In addition, A. 398
aegypti disappeared from several sites by 2005, 399
where it was detected in previous years. A. aegypti 400
not only decreased in prevalence on ovitraps from 401
2003 to 2005, but also its presence across the islands. 402
Interestingly, A. aegypti was rarely present as the sole 403
species in a sample; in those where A. aegypti was 404
detected, at least 98% of the time the traps were also 405
positive for A. albopictus. Considering the total 406
positive samples processed for all 3 years, just 407
Fig. 2 Estimation of oviposition activity by area for years 7.08% contained A. aegypti occurring with A. albo- 408
2000–2007 based on yearly oviposition totals in ovitraps. pictus, and 1.67% of the total positive samples 409
Values should be considered as relative indicators for both contained A. aegypti only; all remaining samples 410
Aedes species (see ‘‘Methods’’)
were positive for A. albopictus only (see Table 1). 411
These results indicate that the two species were 412
368 numbers than the other quartiles (Van der Waerden overlapping throughout the island, possibly occupy- 413
369 v2appr ¼ 29:32, P \ 0.001), while distance to the ing similar habitats. 414
370 nearest building was not associated with the number The expected decline in the frequency of A. aegypti 415
371 of eggs collected by ovitraps (Van der Waerden suggests a rapid, abrupt decline in its presence in the 416
372 v2appr ¼ 4:7, P \ 0.05). islands during the years 2000–2002, consistent with 417
the observed data points (Fig. 5) and observations of 418
373 Temporal patterns vector control officers in the field (David Kendell 419
pers. obs.). This decline occurred during the same 420
374 Egg counts per week reveal a distinct seasonal pattern years when the average number of eggs per ovitrap 421
375 of abundance (Fig. 3). This pattern suggests an early sharply increased (Kruskal–Wallis v2appr ¼ 106:26, 422
376 generation, with major reproduction in late June, P \ 0.001). A similar trend can be observed in the 423
377 followed by at least two more distinct peaks, in proportion of positive ovitraps, as they increased in 424
378 August and September. These latter peaks may the years 2000–2002, to reach stable levels in the 425
379 represent subsequent generations. Although activity years 2002–2007. This increase was not detectable 426
380 declines markedly during the cooler months, eggs with the nonparametric ANOVA (Kruskal–Wallis 427

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Fig. 3 Weekly egg

production, pooled across
years (2002–2007). Mean
number of eggs per trap,
±SE (based on residual
mean square from 1-way
ANOVA)
Author Proof

Table 1 Summary of samples processed, successfully ampli-

fied, and corresponding species identification from Bermuda
samples for years 2003–2005
Year Nt Success A. A. Both
albopictus aegypti species
Ns %s Nal %al Nae %ae Nb %b

2003 138 56 40.6 49 87.5 1 1.79 5 8.93

2004 154 102 66.2 88 86.3 2 1.96 12 11.80
2005 98 82 83.7 79 96.3 1 1.22 2 2.44
All years 390 240 61.5 216 90.0 4 1.67 19 7.08
Fig. 4 Temporal correlation in the number of eggs per ovitrap
in function of yearly time interval. Only CRH significant Nt, total number of samples processed; Success, samples
correlation values after Dutilleul’s correction are used in the successfully amplified; Ns, number of samples successfully
correlation analysis (R2 = 0.5, P \ 0.001) amplified; %s, percentage of samples successfully amplified.
Nal, number of samples only positive for A. albopictus;
428 v2appr ¼ 8:64, P [ 0.05), but it could be detected with %al, percentage of samples only positive for A. albopictus;
Nae, number of samples only positive for A. aegypti;
429 non-overlapping confidence intervals after a bootstrap %ae, percentage of samples only positive for A. aegypti; Nb,
430 procedure. This overall trend presented in Fig. 4a, b number of samples positive for both species; %b, percentage of
431 suggests that the decline in A. aegypti occurs at the samples positive for both species
432 same time that A. albopictus increased dramatically in
433 the years 2002–2007. evidence that suggests a high rate of habitat turnover 441
(Fig. 4), which would reduce the impact of compe- 442
tition, (2) collapse of A. aegypti well in advance of 443
434 Discussion any indication that the two mosquito populations had 444
saturated their environment (Fig. 5), and (3) evidence 445
435 The key issue that we sought to address in this study that the two species do not display substantial 446
436 was the importance of competition in the interaction asymmetry in their resource use or in their compet- 447
437 between two invading species. Our interpretation of itive impacts on one another (Table 2). We argue 448
438 these data raises some concerns that larval competi- below that additional factors are well worth consid- 449
439 tion alone may not account for the extremely rapid ering, including differential susceptibility to parast- 450
440 disappearance of A. aegypti. We base this view on (1) itism, and differential colonizing ability by the two 451

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Fig. 5 Left axis predicted

decline of A. aegypti (dotted
line) based on a fitted
ordinal logistic function
regression for years
1999–2007, solid triangles
depict observed frequency
values. a Right axis average
number of eggs per ovitrap
for years 2000–2007 (solid
line). Error bars represent
standard errors. Letters
Author Proof

indicate overlapping 95%

confidence intervals after
1,000 bootstrap replicates.
b Right axis frequency of
positive ovitraps over totals
for years 2000–2007 (solid
line), error bars represent
standard errors. Similar
letters indicate overlapping
95% confidence interval
after 1,000 bootstrap
replicates

Table 2 Responses of per capita growth rate estimates for each species to the initial density of Aedes aegypti and A. albopictus
larval cohorts in tire fluid, grown under field conditions in Vero Beach, FL (Livdahl 1993)
Species n rmi b1 b2 Kj aij

A. aegypti 49 0.0299 ± 0.0072 -0.0004 ± 0.0002 -0.00005 ± 0.00007 299 0.120

A. albopictus 50 0.0348 ± 0.0069 -0.0005 ± 0.0001 -0.00006 ± 0.00008 278 0.125
Regressions are based on the Lotka-Volterra growth equation for each species, ri0  NdNi dti ¼ rmi þ bi Ni þ bj Nj in which each species has
a maximum per capita rate at minimal density, rmi, and the coefficients bi and bj correspond to the response to conspecific (-rmi/Ki)
and heterospecific (-rmiaij/Ki) densities, respectively. This model assumes linear decline in r0 i as density of either species increases.
Per capita growth rate estimates r0 i were obtained for each species within each replicate tire segment by the method of Livdahl and
Sugihara (1984). Standard errors for each regression term are shown. Only the conspecific regression coefficients (b1) are
significantly different from zero (P \ 0.001). Units are as follows: rm, days-1; b1 and b2, days-1 9 (individuals/250 ml)-1. The
number of r0 values obtained for each regression (n) differ because some cohorts produced no survivors and those replicates were
randomly conjoined with replicates that did yield survivors. Carrying capacities are estimated by Ki = -bi/rmi, shown in individuals
per litre, and competition coefficients by aij = b2/b1. Competition coefficients are dimensionless

452 species in the face of a dynamic landscape of mostly officers’ observation that human activities could be 456
453 domestic habitats. helping the spread of Aedes across the islands. 457
454 The rapid spread of Aedes mosquitoes across the Movement and exchange are commonplace for 458
455 islands (Fig. 2) is consistent with Bermuda health various containers, including potential habitats 459

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460 ranging from ornamental bromeliads to construction- An abrupt decline of one species while its 509
461 related water drums, domestic trash, moored or competitor is at low densities is possible under 510
462 landed boats, and other elements suitable for Aedes classical theory if the carrying capacities of the two 511
463 breeding. The lack of an association between ovipo- competitors are markedly different (asymmetric 512
464 sition activity and distance to buildings differentiates competition, sensu Lawton and Hassell 1981). Juli- 513
465 the existing mosquito-road association from a mere ano (1998) reared these species at very high densities 514
466 human density factor, considering that both are of larvae within plastic and nylon enclosures (approx- 515
467 interrelated. The mosquito-road association suggests imately 200 and 600 initial larvae per litre) in filtered 516
468 that disposal of domestic elements such as cups, cans, tire water with or without fresh leaf litter, and 517
Author Proof

469 or bottles could be helping mosquitoes, providing observed superior success by A. albopictus at high 518
470 both new breeding habitats and opportunities for density with no leaf litter added. The results suggest a 519
471 dispersal. disparity of K values, although K was not estimated. 520
472 Local variation in abundance and frequent shifts in The larval enclosures for this experiment were 521
473 abundance peaks (darker pixels in maps, Fig. 2) vertically oriented, i.e., relatively deep and narrow. 522
474 among years could be attributed to variation in However, Livdahl (1993) grew the same species in 523
475 habitat availability and quality. The decrease in tire segments under field conditions, using unfiltered 524
476 temporal correlation through time (Fig. 4), supports tire water with its debris. Lower densities were used 525
477 this interpretation and suggests a dynamic spatial and than in Juliano’s study (120–480 larvae per litre), and 526
478 temporal variation in local mosquito populations. larvae were free to browse throughout the 250 ml of 527
479 Constant, fine scale extinction and colonization fluid contained within each tire segment. Larval 528
480 events likely occur across the islands, as breeding habitats in this experiment were horizontal in orien- 529
481 habitats are continuously created and destroyed. tation i.e., relatively shallow and broad. Regressions 530
482 Control officers’ responses and citizens’ awareness of per capita growth rate estimates for these species 531
483 of high mosquito densities are most likely a major (summarized in Table 2) yielded rather similar car- 532
484 factor reducing local mosquito populations and rying capacities for the two species, and neither 533
485 producing the observed displacements of high density species had a significant impact on the other’s 534
486 peaks. success. Estimates of competition coefficients (a) 535
487 Considering the previous success of A. aegypti in were similar, although the error in the parameter 536
488 the Bermuda Islands, both in historic times (during estimates needed to calculate a around these esti- 537
489 which it was responsible for numerous yellow fever mates is sufficient to leave much room for doubt. 538
490 epidemics and a dengue outbreaks), and in the period Perhaps the physical heterogeneity of unmodified tire 539
491 1997–2000, intrinsic environmental variables can water enabled the growth of various kinds of 540
492 probably be disregarded as causes for its disappear- microbial resources that the two species could exploit 541
493 ance; most likely, the decline relates to the differentially. If they do exploit different resources as 542
494 A. albopictus introduction in 2000. At first glance, larvae (and Juliano’s results do show a much 543
495 the observed decline in A. aegypti, concurrent with greater need for leaf litter by A. aegypti than by 544
496 the increase in A. albopictus numbers, appears A. albopictus), then we would not expect such a rapid 545
497 consistent with a competitive exclusion scenario. decline of A. aegypti as we have seen in the present 546
498 The decrease in A. aegypti occurred at the same time study (Fig. 5). 547
499 that the total numbers of eggs sharply increased The disparity between the results of those two 548
500 during the period 2000–2002. The total number of competition studies, which were conducted concur- 549
501 eggs leveled off in the period 2002–2007, which rently in the same locality, illustrates both the 550
502 could suggest that A. albopictus had taken over all potential variability in competitive intensity that 551
503 breeding habitats, reaching carrying capacity (Fig. 5). could be faced by larvae occupying a wide variety 552
504 However, if competitive exclusion between two of habitat conditions, as well as the difficulty in 553
505 similar species takes place following the classical interpreting population dynamics from the restricted 554
506 competition theory, it could be expected to occur perspective of a larval competition experiment. 555
507 when the superior competitor is close to its equilib- Certainly, the conditions faced by larvae of both 556
508 rium density (see Fig. 1a, b). species would vary much more in field populations, 557

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558 which may occupy containers ranging in size from deleterious effects (Munstermann and Wesson 1990), 607
559 bottle caps to cisterns. and others do not (Blackmore et al. 1995 found 608
560 In sum, we consider that a rapid competitive higher prevalence of Ascogregarina spp. among A. 609
561 exclusion scenario such as represented in Fig. 1c, d is aegypti in areas that had not yet been invaded by A. 610
562 unlikely after the following considerations. The per albopictus), but the Ascogregarina-Aedes interactions 611
563 capita growth rate and K estimations for both species have still not been thoroughly studied and a number 612
564 are similar (Table 2), which should not allow the of other parasites, including fungi, flagellates and 613
565 observed fast decline of A. aegypti (Fig. 5). In bacteria could also play a role (Fukuda 1992). 614
566 addition, A. albopictus population only appears to Without direct examination of the full array of 615
Author Proof

567 have leveled off after A. aegypti’s acute decrease, potential parasites, it is difficult to exclude parasitism 616
568 reaching much higher levels in later years, suggesting as a contributing factor because a parasite may 617
569 that it was not close to its equilibrium density (K), a impose effects on its hosts similar to those that could 618
570 condition for competitive exclusion of species that be expected from resource competition. 619
571 have similar competitive parameters (see Fig. 1). Mediation of competition by climate has been 620
572 Furthermore, the intensity of competition between suggested as a mechanism for coexistence between 621
573 these two species appears to vary substantially with these species, which has been observed after periods 622
574 different experimental conditions (e.g., Black et al. of drought (Britch et al. 2008) and microclimates 623
575 1989; Juliano 1998, 2007; Livdahl 1993) which that favor the more desiccation resistant eggs of 624
576 suggests that in highly heterogeneous urban environ- A. aegypti (Juliano et al. 2002). Climate trends could 625
577 ments, such as in Bermuda where multiple containers also accelerate the competitive exclusion process. 626
578 types are available, and with constant turnover of A. aegypti may be near the limits of its ecological 627
579 habitats, each species could be able to prevail in range in terms of tolerance to winter conditions in 628
580 certain types. Following these considerations, it subtropical habitats such as Florida and Bermuda. 629
581 appears that an explanation based on larval resource Figure 3 indicates a dramatic decline in activity 630
582 competition alone is incomplete. Perhaps classical during Bermuda’s winter, and A. albopictus, which 631
583 theory does not describe accurately the present most likely has origins in temperate Asia, may have 632
584 situation, e.g., nonlinear responses to conspecific or an advantage under such conditions that could 633
585 interspecific density could exist, or other processes accelerate the displacement of A. aegypti. 634
586 besides competition should be reconsidered. None of these alternatives should be considered as 635
587 Several additional considerations could be made; an exclusive explanation; potential synergistic effects 636
588 one possibility is that A. albopictus introduced with competition need to be taken in consideration. 637
589 diseases to the islands that compromise A. aegypti These results fuel interesting questions and invite 638
590 populations (such as Ascogregarina, e.g., Blackmore further studies in this system that could help clarify 639
591 et al. 1995), causing a reduction in their rate of the mechanism of displacement. Competition exper- 640
592 growth. Juliano (1998) found no evidence to support iments between A. albopictus and A. aegypti per- 641
593 this idea in his Florida competition study, but he did formed in the field would allow a test of the 642
594 not manipulate the presence or absence of parasites to competitive exclusion hypothesis and the appropri- 643
595 examine relative susceptibilities, and did not examine ateness of the classical theory for this system. 644
596 dead larvae from his competition experiment, in However, given the variety of different sorts and 645
597 which some treatment combinations resulted in total sizes of containers used by these species, it would be 646
598 larval mortality. The potential for parasite alteration difficult to conduct an experiment that encompasses 647
599 of a competitive interaction has been shown by the full diversity of habitats, and it is always possible 648
600 Aliabadi and Juliano (2002), who showed a reduced to force species to compete in an experimental 649
601 dominance by A. albopictus when competing with setting. In addition, the A. albopictus population in 650
602 native A. triseriatus. The degree of host specificity the islands could be screened with molecular methods 651
603 and pathogenicity by the two species of Ascogrega- for microparasites known to infect A. aegypti. The 652
604 rina involved remains uncertain; some studies indi- investigation by health department officers of loca- 653
605 cate that cross-infection between these two tions of very early detections of A. albopictus 654
606 mosquitoes and their natural parasites can occur with suggested a single entry in the islands associated 655

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656 with a single bromeliad shipment (D. Kendell, pers. Aedes albopictus and Aedes aegypti (Diptera: Culicidae). 706
657 obs.). If a very limited number of individuals are J Med Entomol 26:260–271 707
Blackmore MS, Scoles GA, Craig GC Jr (1995) Parasitism of 708
658 responsible for A. albopictus’ invasion in Bermuda, it Aedes aegypti and Aedes albopictus (Diptera: Culicidae) 709
659 seems likely that they did not carry the complete load by Ascogregarina spp. (Apicomplexa: Lecudinidae) in 710
660 of microparasites that otherwise could be found Florida. J Med Entomol 32:847–852 711
661 associated with A. albopictus in other regions of Boromisa RD, Rai KS, Grimstad PR (1987) Variation in the 712
vector competence of geographic strains of Aedes albo- 713
662 the world. We have recently identified Ascogregari- pictus for dengue 1 virus. J Am Mosq Control Assoc 714
663 na taiwanensis in Bermuda A. albopictus larvae 3:378–386 715
664 (J. Sohighian and T. Livdahl, unpublished), so Braks MAH, Honório NA, Lourenço-de-Oliveira SA, Juliano 716
SA, Lounibos LP (2003) Convergent habitat segregation 717
Author Proof

665 A. albopictus’ success cannot be attributed to an

of Aedes aegypti and Aedes albopictus (Diptera: Culici- 718
666 escape from parasitism, although there could be other dae) in southeastern Brazil and Florida. J Med Entomol 719
667 parasites, such as microsporidians (Andreadis 2007), 40:785–794 720
668 that may interact with these species and alter the Braks MAH, Honório NA, Lounibos LP, Lourenço-De-Oli- 721
669 outcome. It is interesting to note that microsporidians veira R, Juliano SA (2004) Interspecific competition 722
between two invasive species of container mosquitoes, 723
670 occur in A. aegypti host populations but have yet to Aedes aegypti and Aedes albopictus (Diptera: Culicidae), 724
671 be discovered in invasive A. albopictus popula- in Brazil. Ann Entomol Soc Am 97:130–139 725
672 tions (T. Andreadis, pers. comm.) associated with Britch SC, Linthicum KJ, Anyamba A, Tucker CJ, Pak EW 726
673 A. albopictus in other regions of the world. (2008) Long-term surveillance data and patterns of inva- 727
sion by Aedes albopictus in Florida. J Am Mosq Control 728
Assoc 24:115–120 729
674 Acknowledgments We thank Ross Furbert and his staff at the Camargo S (1967) History of Aedes aegypti eradication in the 730
675 Vector Control unit, Bermuda Ministry of Health, for their kind Americas. Bull WHO 36:602–603 731
676 cooperation and assistance. The Bermuda Institute of Ocean Castro Gomes A, Souza JP, Bergamaschia DP, dos Santos JF, 732
677 Sciences (BIOS) provided generous assistance to L. Kaplan Roberto Andrade VR, Leiteb OF, Rangel O, Souza SL, 733
678 travel to the site. Personnel at the Florida Medical Entomology Guimarães NN, Limab VC (2005) Anthropophilic 734
679 Laboratory, especially G. F. O’Meara and L. P. Lounibos, activity of Aedes aegypti and of Aedes albopictus in area 735
680 provided valuable assistance during the establishment and under control and surveillance. Rev Saúde Pública 736
681 maintenance of the competition experiment described in 39:1–5 737
682 Table 2, and that research was supported by a grant from the Clark Labs (2007) Idrisi Los Andes edition v.15.0. Worcester 738
683 Florida Department of Agriculture and Consumer Services, Diallo M, Ba Y, Faye O, Soumare ML, Dia I, Sall AA (2008) 739
684 Bureau of Entomology and Pest Control to T. Livdahl. We Vector competence of Aedes aegypti populations from 740
685 thank one anonymous reviewer and D. Simberloff for useful Senegal for sylvatic and epidemic dengue 2 virus isolated 741
686 comments on the manuscript, and Mark Berman for his in West Africa. Trans R Soc Trop Med Hyg 102:493–498 742
687 assistance with the Florida experiment. This research was ESRI (2006) ArcMAP version 9.2. Redlands 743
688 supported by the National Institutes of Health (USA) (grant 1 Fontenille D, Rodhain F (1989) Biology and distribution of 744
689 R15 AI062712-01 to T. Livdahl) and the Keck Foundation. Aedes albopictus and Aedes aegypti in Madagascar. J Am 745
Mosq Control Assoc 5:219–225 746
Francy DB, Moore CG, Eliason DA (1990) Past, present and 747
future of Aedes albopictus in the United States. J Am 748
Mosq Control Assoc 6:127–132 749
690 References Fukuda T (1992) A preliminary report of a survey for parasites 750
and pathogens of Aedes albopictus and other container- 751
691 Aliabadi BW, Juliano SA (2002) Escape from gregarine par- inhabiting mosquitoes in North Central Florida. J Fla 752
692 asites affects the competitive interactions of an invasive Mosq Control Assoc 63:76–77 753
693 mosquito. Biol Inv 4:283–297 Gilotra SK, Rozeboom LE, Bhattacharya NC (1967) Obser- 754
694 Andreadis TG (2007) Microsporidian parasites of mosquitoes. vations on possible competitive displacement between 755
695 J Am Mosq Control Assoc 23(sp2):3–29 populations of Aedes aegypti linnaeus and Aedes albo- 756
696 Beaty J, Aitken THG (1979) In vitro transmission of yellow pictus skuse in Calcutta. Bull WHO 37:437–446 757
697 fever virus by geographic strains of Aedes aegypti. Mosq Griswold MW, Lounibos LP (2006) Predator identity and addi- 758
698 News 39:232–238 tive effects in a treehole community. Ecology 87:987–995 759
699 Benedict MQ, Levine RS, Hawley WA, Lounibos LP (2007) Gubler DJ (1998) Dengue and dengue hemorrhagic fever. Clin 760
700 Spread of the tiger: global risk of invasion by the mos- Micro Rev 11:480–496 761
701 quito Aedes albopictus. Vector-Borne Zoonot Dis 7:76–85 Hawley WA (1988) The biology of Aedes albopictus. J Am 762
702 Beyer HL (2004) Hawth’s analysis tools for ArcGIS. Available Mosq Control Assoc 4(sp1):1–40 763
703 at http://www.spatialecology.com/htools Hidayati H, Mohd SA, Nazni WA, Lee HL (2005) Insecticide 764
704 Black WC, Rai KS, Turco BJ, Arroyo DC (1989) Laboratory resistance development in Culex quinquefasciatus (Say), 765
705 study of competition between United States strains of Aedes aegypti (L.) and Aedes albopictus (Skuse) larvae 766

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Article No. : 9721 h LE h TYPESET
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h 4 DISK
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 L. Kaplan et al.

767 against malathion, permethrin and temephos. Trop Bio- Munstermann LE, Wesson DM (1990) First record of Asco- 812
768 med 22:45–52 gregarina taiwanensis (Apicomplexa: Lecudinidae) in 813
769 Hornby JA, Moore DE, Miller TW Jr (1994) Aedes albopictus North American Aedes albopictus. J Am Mosq Control 814
770 distribution, abundance, and colonization in Lee County, Assoc 6:235–243 815
771 Florida, and its effect on Aedes aegypti. J Am Mosq Murrell EG, Juliano SA (2008) Detritus type alters the outcome 816
772 Control Assoc 10:397–402 of interspecific competition between Aedes aegypti and 817
773 Insightful Corp (2007) S-plus v.8.0.4. Palo Alto Aedes albopictus (Diptera: Culicidae). J Med Entomol 818
774 Juliano SA (1998) Species introduction and replacement 45:375–383 819
775 among mosquitoes: interspecific resource competition or Novak RJ (1992) The Asian tiger mosquito, Aedes albopictus. 820
776 apparent competition? Ecology 79:255–268 Wing Beats 3:5 821
777 Juliano SA (2007) Species interactions among larval mosqui- O’Meara GF, Gettman AD, Evans LF Jr, Scheel FD (1992) 822
778 toes: context dependence across habitat gradients. Annu Invasion of cemeteries in Florida by Aedes albopictus. J 823
Author Proof

779 Rev Entomol 54:37–56 Am Mosq Control Assoc 8:1–10 824

780 Juliano SA, O’Meara GF, Morrill JR, Cutwa MM (2002) O’Meara GF, Evans LF Jr, Gettman AD, Cuda JP (1995) 825
781 Desiccation and thermal tolerance of eggs and the coex- Spread of Aedes albopictus and decline of Ae. aegypti 826
782 istence of competing mosquitoes. Oecologia 130:458–469 (Diptera: Culicidae) in Florida. J Med Entomol 32: 827
783 Juliano SA, Lounibos LP, Nishimure N, Greene K (2009) Your 554–562 828
784 worst enemy could be your best friend: predator contri- Pumpuni CB, Kneples J, Craig G (1992) Influence of temper- 829
785 butions to invasion resistance and persistence of natives. ature and larval nutrition on the diapause inducting pho- 830
786 Oecologia Online publication date: 16 Oct 2009 toperiod of Aedes albopictus. J Am Mosq Control Assoc 831
787 Kesavaraju B, Damal K, Juliano SA (2008) Do natural con- 8:223–227 832
788 tainer habitats impede invader dominance? Predator- Rawlins SC, Martinez R, Wiltshire S, Legall G (1998) A 833
789 mediated coexistence of invasive and native container- comparison of surveillance systems for the dengue vector 834
790 dwelling mosquitoes. Oecologia 155:631–639 Aedes aegypti in Port of Spain, Trinidad. J Am Mosq 835
791 Kobayashi M, Nihei N, Kurihara T (2002) Analysis of northern Control Assoc 14:131–136 836
792 distribution of Aedes albopictus in Japan by geographical Rosenberg MS (2001) PASSAGE. Pattern analysis, spatial 837
793 information system. J Med Entomol 39:4–11 statistics, and geographic exegesis. Version 1.0. Tempe 838
794 Livdahl T (1993) Competition and egg-hatch inhibition among SAS Institute Inc (2007) JMP version 7.0. Cary 839
795 tire-breeding mosquitoes. Technical report, Florida Stasiak RS, Grothaus RH, Miner WF (1969) Susceptibility of 840
796 Department of Agriculture and Consumer Services, Aedes albopictus (skuse) larvae from South Vietnam to 841
797 Bureau of Entomology and Pest Control, Jacksonville, FL five insecticides, 1969. Mosq News 30:246–249 842
798 Livdahl T, Willey MS (1991) Prospects for an invasion: Tabachnick WJ (1991) Evolutionary genetics and arthropod- 843
799 competition between Aedes albopictus and native Aedes borne disease. The yellow fever mosquito. Am Entomol 844
800 triseriatus. Science 253:189–191 37:14–24 845
801 Lounibos LP (2002) Invasions by insect vectors of human Urbanelli S, Bellini R, Carrieri M, Salicandro P, Celli G (2000) 846
802 disease. Annu Rev Entomol 47:233–266 Population structure of Aedes albopictus (Skuse): the 847
803 Lounibos LP (2007) Competitive displacement and reduction. mosquito which is colonizing Mediterranean countries. 848
804 AMCA Bull 23(sp):276–282 Heredity 84:331–337 849
805 Mitchell CJ (1995) The role of Aedes albopictus as an arbo- Wesson DM (1990) Susceptibility to organophosphate insec- 850
806 virus vector. Parasitology 37:109–113 ticides in larval Aedes albopictus. J Am Mosq Control 851
807 Moore PR, Johnson PH, Smith GA, Ritchie SA, Van Den Hurk Assoc 6:258–264 852
808 AF (2007) Infection and dissemination of dengue virus Wesson DM, Porter CH, Collins FH (1992) Sequence and 853
809 type 2 in Aedes aegypti, Aedes albopictus, and Aedes secondary structure comparisons of ITS rDNA in mos- 854
810 scutellaris from the Torres Strait, Australia. J Am Mosq quitoes (Diptera: Culicidae). Mol Phyl Evol 1:253–269 855
811 Control Assoc 23:383–388 856

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