2008 - Dujardin - Morphometrics Applied To Mediacal Entomology

Infection, Genetics and Evolution 8 (2008) 875890
Contents lists available at ScienceDirect
Infection, Genetics and Evolution

journal homepage: www.elsevier.com/locate/meegid
Morphometrics applied to medical entomology

Jean-Pierre Dujardin a,b
a b
UMR IRD - CNRS 2724, Agropolis, 911, BP 64501, 34394, Montpellier, France CVVD, Faculty of Science, Mahidol University, Bangkok, Thailand
A R T I C L E I N F O
A B S T R A C T
Article history: Received 20 March 2008 Received in revised form 28 July 2008 Accepted 30 July 2008 Available online 11 September 2008 Keywords: Geometric morphometrics Insects vectors Cryptic species Population structure Reinfesting insects
Morphometrics underwent a revolution more than one decade ago. In the modern morphometrics, the estimate of size is now contained in a single variable reecting variation in many directions, as many as there are landmarks under study, and shape is dened as their relative positions after correcting for size, position and orientation. With these informative data, and the corresponding software freely available to conduct complex analyses, signicant biological and epidemiological features can be quantied more accurately. We discuss the evolutionary signicance of the environmental impact on metric variability, mentioning the importance of concepts like genetic assimilation, genetic accommodation, and epigenetics. We provide examples of measuring the effect of selection on metric variation by comparing (unpublished) Qst values with corresponding (published) Fst. The primary needs of medical entomologists are to distinguish species, especially cryptic species, and to detect them where they are not expected. We explain how geometric morphometrics could apply to these questions, and where there are deciencies preventing the approach from being utilized at its maximum potential. Medical entomologists in connection with control programs aim to identify isolated populations where the risk of reinfestation after treatment would be low (biogeographical islands). Identifying them can be obtained from estimating the number of migrants per generation. Direct assessment of movement remains the most valid approach, but it scores active movement only. Genetic methods estimating gene ow levels among interbreeding populations are commonly used, but gene ow does not necessarily mean the current ow of migrants. Methods using the morphometric variation are neither suited to evaluate gene ow, nor are they adapted to estimate the ow of migrants. They may provide, however, the information needed to create a preliminary map pointing to relevant areas where one could invest in using molecular machinery. In case of reinfesting specimens after treatment, the question relates to the likely source of reinfesting specimens: are they a residual sample not affected by the control measures, or are they individuals migrating from surrounding, untreated foci? We explain why the morphometric approach may be adapted to answer such question. Thus, we describe the differences between estimating the ow of migrants and identifying the source of reinfestation after treatment: although morphometrics is not suited to deal with the former, it may be an appropriate tool to address the latter. 2008 Elsevier B.V. All rights reserved.
1. Introduction For medically important insects submitted to various control pressures and landscape changes, phenotypic variation is frequently observed and is the source of many taxonomic and epidemiological questions. Nevertheless, phenotypic variation of medically impor-
E-mail address: dujardinbe@gmail.com. 1567-1348/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.meegid.2008.07.011
tant insects is seldom measured. Perhaps it does not receive adequate consideration because it is often believed that phenotypic variation is the transient reection of environmental variation. In species detection (see Section 4) and reinfestation studies (see Section 5.1), morphometrics takes advantage of the environmental impact on phenotypic variation: however, more is involved than just the environment. There is indeed a growing body of evidence that phenotypic plasticity has evolutionary importance (Pigliucci, 2005). As long as phenotypic variation has environmental and/or genetic
876
J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890
Fig. 1. Screenshot of the COO software (see Section 7) showing landmarks taken on the wing of Triatoma protracta. Circles indicating each landmark were added on the screenshot picture.
causes, its study can help medical entomologists to detect local populations with potentially important characters. We believe that the rst step toward a better understanding of the epidemiological importance of phenotypic variation is to implement adequate quantication. Morphometrics, particularly geometric morphometrics, is a relevant tool for this type of task. It focuses on phenotypic variation, its parametrization and its relation to extrinsic factors. It is helpful to biologists working with any organisms that can be characterized by a few landmarks, including parasites (Hugot and Baylac, 1996; Periago et al., 2008). 1.1. Tradition and modernity Morphometrics is often presented as traditional, making use of limited sets of measurements, versus modern (or geometric), making use of total geometric information. This distinction refers mainly to the type of metric data. Morphometric characters are related to growth and development, they are continuous characters. Traditionally, they were estimates of distances between anatomical points called landmarks. Since the 1990s they have come to be the coordinates of these landmarks in a given system of orthogonal axes (Fig. 1). This difference has important implications (Rohlf and Marcus, 1993; Adams et al., 2004). The most important one is the increase in information embedded in the new kind of data: the coordinates contain not only size, such as distances between landmarks, but also shape, such as their relative position. The most spectacular application is the visualization of shape itself by means of informatic techniques; and the visualization of its variation among individuals or groups. Despite these advantages and the attractiveness of modern morphometrics, it has yet to gain popularity in medical entomology, where traditions may be hard to dislodge.
1.2. Size and shape Morphometric techniques measure size, shape and the relation between size and shape (allometry). In practice, size and shape refer to a measurable part of the organism under study, not the complete organism. Moreover, only a fraction of this part is measured. On insects, wings are usually preferred to any other organ because of its almost two-dimensional structure reducing digitizing error. A few landmarks available on a wing do not completely describe the wing, but the objective of morphometrics is not to provide a complete description of an organisms dimensions, but rather to allow valid comparisons of a given phenotype among individuals, local populations and/or species. Provided there is operational1 homology among individual landmarks, only a partial capture of size and shape is needed. 1.2.1. Size Size is easy to conceive when only two landmarks are considered: it is the distance between them. When more landmarks are collected, the use of distances between each of them creates many variables so that the relevant information may become difcult to extract. To avoid such problems, traditional entomologists have often selected one dimension as reecting the global size of the insect. For instance, the length of the wing along
1 In a broad sense, homology is correspondance of parts with no specication about whether the parts correspond with respect to structure, development, or phylogeny. Operational homology refers to anatomical correspondance of parts, without proof of similarity by common descent. If individuals correspond to a single species, homologous landmarks are probably similar due to common descent because all members of the species come from a common ancestor. If they correspond to different species, there is no guaranty that homologous landmarks are similar due to common descent, except if they are known to be descending from a common ancestor (Lele and Richtsmeyer, 2001).
877
Fig. 2. Centroid size of the wing of Triatoma protracta using the landmarks shown in Fig. 1. There is a central point (the centroid) which is located at the average coordinates of all the landmarks, from which a distance is computed to each landmark (see white lines). The square root of the sum of these squared distances is used as the centroid size.
Fig. 3. Shape comparison between three geographic populations of Triatoma protracta using the landmarks shown in Fig. 1, using software MOG (see Section 7). Each conguration is the average conguration of the corresponding group: T. p. protracta laying rough eggs, T. p. p. laying smooth eggs and T. p. woodi. Each circle represents a landmark; at each position, one should see only one circle. Lack of perfect superposition means shape difference, which can then be located on the wing. Procrustes distance is derived from these differences at each landmark.
its largest axis is frequently used as an estimator of the global size of the insect (Garcia et al., 2001; Lehmann et al., 2006). Such relationships are often assumed instead of demonstrated (Siegel and Novak, 1992), but here the point is that only a single dimension, the length of the largest axis of the wing, is considered. The size of the wing could change along an axis other than the largest one, which is completely ignored by this approach. What is needed is (i) an estimation of size that can detect changes in any direction, not only along the largest axis, and, preferably, (ii) an overall estimation of size, at least the size of the anatomical structure under study. This is provided nicely by the centroid size2 in geometric morphometrics (Fig. 2). For instance, the centroid size of the wing of Aedes aegypti was amplied by either lower larval densities or higher larval food concentrations (Jirakanjanakit et al., 2007). Were the same landmarks affected by the same displacements? The answer is no: the posterior border of the wing showed opposite movement depending on the external stimulus, either food or density (Jirakanjanakit et al., 2007). 1.2.2. Shape Not only in medical entomology, but also in many elds where morphometrics is applied, shape has been traditionally described as the ratio of one dimension to another. Although the ratio might appear, intuitively, able to scale for size, it often does not, and this is simple to demonstrate (Dujardin and Slice, 2007). Moreover, the using ratios introduce some well-known statistical drawbacks (Albrecht et al., 1993). Angles do not improve the situation since they are another kind of ratio (Burnaby, 1966). In geometric morphometrics, the shape of a conguration of landmarks is represented by their relative positions as contained in their coordinates. Coordinates are able to reproduce the shape and its variation among individuals, allowing a direct visualization of metric changes. To compare the shape of different congurations, raw coordinates must be corrected so that they can be properly superimposed, including by scaling for size. This procedure is called Procrustes superimposition (Rohlf, 1990)3: it produces
2 Centroid size is the squareroot of the sum of squared distances between centroid and each landmark (see Fig. 2). 3 Procrustes, whose name means he who stretches, was a thief in Greek mythology (the myth of Theseus). He preyed on travelers along the road to Athens. He offered his victims hospitality on a magical bed that would t any guest. As soon as the guest lay down Procrustes went to work upon him, either stretching the guest or cutting off his limbs to make him t perfectly onto the bed (Grose Educational Media, 19971998).
residual coordinates (Fig. 3). These can be considered as shape variables, but they lost degrees of freedom and need further modications (producing the so called partial warps4) so that they can be studied using classical statistical techniques (Rohlf and Bookstein, 2003). 1.2.3. Allometry Since each form can be explained by the change in linear dimensions, it is obvious that size and shape are not independent attributes. The relationship between them is called allometry, which can be evaluated by linear regression techniques. Under certain circumstances, the residuals from this regression analysis may be used to produce allometry-free variables. This approach for size scaling in traditional morphometrics is more accurate than the use of ratios (for a review, see Klingenberg (1996)). In geometric morphometrics the removal of allometric residues is seldom needed.5 The correlation between size and shape variables can be high, as in shape changes due to growth, but it can also be very low or even null, as is sometimes observed in changes caused by some evolutionary diverging processes. In studies comparing insects belonging to the same species, size variation is often an environmentally induced and reversible character: one should then verify whether shape variation is not simply the passive consequence of size variation.
2. From dimensions to biology The mechanisms underlying size or shape variation either a larger number of cells, or larger cells and the corresponding hormonal triggers are the subject of important studies (Nijhout, 2003; Davidowitz et al., 2004). However, from an epidemiological point of view, it is not directly important to know these mechanisms, but rather the association between, say, larger size and higher transmission rate, is the relevant character. It is the
4 The coordinates after Procrustes superimposition have lost four degrees of freedom (in two-dimensional congurations) and lie in a non-Euclidean, curved space (Kendall, 1986): as such, they would not be amenable to multivariate analyses (Rohlf, 1996); projection techniques onto an Euclidean space tangent to the curved space produce the so-called partial warps which may be submitted to standard multivariate analyses. 5 Residual shape coordinates after Procrustes superimposition are scaled for the isometric change of size, not the allometric one. Allometric variation is still embedded in the shape variables, and to remove it can be desirable in some analyses.
878
commonly observed association in a given species in a given area that is the indicator by which a population or an area is put under special epidemiological surveillance. These associations between size and other traits of the insect raised many questions about size as an indicator of tness. Size, understood as the global size of the insect, has been associated with many tness components in medically important mosquitoes (see hereunder) and other insects (Honek, 1993). In mosquitoes, various levels of correlation, often a positive correlation, have been found with fecundity (Lyimo and Takken, 1993; Blackmore and Lord, 2000), parity and survivorship (Haramis, 1983; Kitthawee et al., 1992; Takken et al., 1998; Manoukis et al., 2006), longevity (Maciel de Freitas et al., 2007), multiple bloodfeeding habits (Xue et al., 1995; Takken et al., 1998), host seeking response (Klowden et al., 1988; Takken et al., 1998), parousness and insemination (Ameneshewa and Service, 1996), resistance to insecticides (Bourguet et al., 2004; Wiwatanaratanabutra and Kittayapong, 2006), dispersal capacity and vectorial competence (Ameneshewa and Service, 1996; Mwangangi et al., 2004; Maciel de Freitas et al., 2007; Schneider et al., 2007). Fewer observations or studies are found on body size and tness in other medically important insects: such as Glossinidae (De Deken et al., 1997), Triatominae (Dujardin et al., 1999f,g), Phlebotominae (larger males in larger leks (Jones and Quinnell, 2002)), or Simuliidae (Malmqvist et al., 2004). Body size could itself be a tness character. For instance, the apparently higher resistance of large insects to some insecticides could be the passive result of the greater surface-to-volume ratio of small insects, increasing their relative exposure to contact insecticides (Johansen, 1972). Whether body size evolved as an adaptive trait, or whether its properties are a by-product of other traits is still an unanswered question. Size often shows high heritability values (Daly, 1992; Lehmann et al., 2006; Dujardin et al., 2007). This is in agreement with the existence of geographic subpopulations that are genetically different for size (Anderson, 1973; Partridge et al., 1994). A high heritability value is not expected for a trait supposedly connected with tness (Mousseau and Roff, 1987). Few laboratory studies controlling for confusive variables have been conducted to validate common observations. For instance, larger size has often been related to longer development times, but experimental work revealed exactly the opposite (Klingenberg and Spence, 1997), or no correlation at all (Caro-Riano et al., unpublished data). Thus, care should be taken not to give size the direct responsibility for increased tness: it could be the passive consequence of another variation, itself directly related to tness. For instance, the reported association of larger size and increased vectorial capacity in Ae. aegypti (Strickman and Kittayapong, 2003) was not conrmed by laboratory experiments (Schneider et al., 2007). It could actually result from the relationship between richer breeding sites and corresponding shorter developmental cycles. Richer larval food and shorter cycles produce both larger size (Klingenberg and Spence, 1997; Jirakanjanakit, 2007) and more abundant insects, the latter effect only being the likely cause of higher transmission rates (Jirakanjanakit, 2007). 2.1. Environmental effects and heritable consequences 2.1.1. Environmental effect on size The environment has a direct and important effect on the size of living organisms, an effect known as Bergmanns rule, which was originally referred to as a general positive correlation between intraspecic body size of endothermic vertebrates and latitude (James, 1970). This rule was extended to ectothermic animals by the studies of Standfuss on Lepidoptera in 1895 (in Atkinson,
1994). Several reviews have veried its validity for most of the ectotherms species studied (Lane and Marshall, 1981; Ray, 1960; Atkinson, 1994). The underlying mechanisms are not always clear, but temperature seems to play a key role (Vanvoorhies, 1996). Experimentally, adult insects generally have a smaller body size when larvae are reared at higher temperatures (Atkinson, 1994). Other environmental factors affect size variation in insects even in the absence of temperature variation. In Ae. aegypti, larval food concentration and/or larval density have been shown to be linearly related to adult size (Jirakanjanakit et al., 2007). The same factors, population density and diet, were shown (Caro-Riano et al., unpublished data) to explain the typical size modication observed in Triatominae in their transition from sylvatic to domestic habitats (Zeledon et al., 1970; Zeledon, 1981; Dujardin et al., 1997a, 1999g; Jaramillo et al., 2002; Rodrguez et al., 2007; Feliciangeli et al., 2007). 2.1.2. Environmental inuence on shape Because environmental change primarily affects the size of the insect, which in turn should produce passive shape changes (allometric effects), it is expected that shape change among geographic areas would be limited to just that: an allometric effect of size change. To verify this hypothesis, using allometry-free variables is recommended: among conspecic populations, there should be no more signicant differences when using allometryfree variables. The question is: Among conspecic geographic populations, is shape variation simply the passive consequence of size variation?. Or alternatively: Is allometry-free shape variation found more commonly or exclusively between species?. The answer seems to be that between conspecic populations allometry-free variation of shape also appears, probably more frequently when ecogeographic differences are signicant ones. This has been observed for instance in Diptera (Baylac and Daufresne, 1996), and the question has been specically addressed for sand ies using traditional morphometric techniques (Dujardin and Le Pont, 2004). It has been shown that size varied among conspecic subpopulations whatever the relative geographic distance or ecoclimatic difference, while shape showed consistent differences only from one large ecoclimatic area to another. Thus, within any one of the ve large ecogeographic regions of South America, the allometry-free properties of sandies showed little variation; a signicant difference appeared only when crossing the different regions. These results were veried among conspecic populations of ten different species (Fig. 5). Allometry-free shape appears as a more stable trait than size, and may require important external changes to be signicantly modied. 2.1.3. Heritable consequences of environmental effects The obvious environmental inuence on size (or shape) among conspecic subpopulations does not justify excluding possible transgenerational effects. As already mentioned, size in insects may show consistent heritability values, so that they can be experimentally selected to constitute subpopulations genetically distinct for size (Anderson, 1973; Partridge et al., 1994). Experimental populations of Drosophila pseudoobscura that were maintained at different temperatures for 6 years diverged so that even when reared at the same temperature, the cold-adapted ies were larger than those that were warm-adapted (Anderson, 1973). This experiment parallels the observation on natural populations of the medically important triatomine bug Rhodnius pallescens (Jaramillo, N., PhD thesis). Five laboratory lines of R. pallescens, some of them reared over more than 20 generations, have been shown to harbor distinct sizes in accordance with the temperature of their region of origin (Fig. 4). The absence of size
879
modation is a generalization of genetic assimilation (Braendle and Flatt, 2006). In this general model, it is argued that environmentally triggered novelties may have greater evolutionary potential than mutationally induced ones, mainly because of two features (Gorur, 2005): (i) they concern populations rather than individuals (while mutations are individual events) and (ii) they represent optimal or close to optimal adaptations (while mutations are often counterselected). With time (the concept of recurrence), these two features increase the likelihood of genetic assimilation, ending up in local genetic changes. 2.1.5. Epigenetics Not only genetic assimilation, but also various aspects of the modern understanding of epigenetic inheritance are reminiscent of Lamarcks ideas on evolution. One of the denitions of epigenetic changes is that they are heritable changes in gene function that occur without a change in the sequence of nuclear DNA (Jablonka and Lamb, 2002). Epigenetic studies focus on how environmental factors affecting a parent can result in changes in the way genes are expressed in the offspring (Jablonka et al., 1992; Bird, 2007). Epigenetic mechanisms such as DNA methylation, histone acetylation (producing changes to the chromatin packaging of DNA), and RNA interference (regulation of gene-expression control by noncoding RNA), and their effects in gene activation and silencing, are increasingly understood to play a role in phenotype transmission and development (Bird, 2007). They provide molecular support to the hypothesis of genetic assimilation. Recent experiment not only reproduced the genetic assimilation experimentally, they also identied the molecular mechanisms. Heritable phenotypic variation can be induced by environmental effects such as the heat shock, involving the action of the heat shock protein Hsp90 (Rutherford and Lindquist, 1998; Debat et al., 2006) or the genes regulating hormonal titers (Suzuki and Nijhout, 2006; Pennisi, 2006), described as capacitors for morphological evolution. The capacitor buffers genotypic variation under normal conditions, thereby promoting the accumulation of hidden polymorphism: genetic mutations can remain masked until the environment (or another mutation) reveals them (Bergman and Siegal, 2003). Recent advances suggest that Hsp90 buffering effects can be trait and/or species specic, limiting the possibility of generalization, and that other such mechanisms are expected to be discovered in the future (Patterson and Klingenberg, 2007). 2.2. Heritable changes and phenotypic consequences Among heritable changes we will mention genetic drift, hybridism and mutation. 2.2.1. Genetic drift as a cause of metric change Population density, food incomes and temperature are known to modify the size of an insect, and, to a lesser extent, its shape (Jirakanjanakit et al., 2007). What about genetic drift? Among conspecic populations, whether they are geographic, ecoclimatic or ecotypic subpopulations, we can expect some degree of genetic drift to occur. Does genetic drift affect primarily size, or shape? In one experiment on isofemale lines of Ae. aegypti, it was shown that the signicant effect was on shape, with non-signicant changes in size (Jirakanjanakit et al., 2008). Moreover, this change produced by genetic drift did not affect the same landmarks as those affected by larval food or density variation (Jirakanjanakit, 2007). 2.2.2. Hybridism Between seven laboratory colonies of Triatoma protracta, or between each of the ve subspecies of T. protracta, each hybrid had an intermediate centroid size between parents (Dujardin et al.,
Fig. 4. Centroid size (ctr) of the wings of Rhodnius pallescens on vertical axis, and mean annual temperatures on horizontal axis corresponding to different Colombian localities (SO, San Onofre, Sucre; GA, Galeras, Sucre; SB, San Bernardo, Cordoba; SC, San Carlos, Antioquia) and to the central region of the Panama Republic (PA). The various colonies of R. pallescens, in spite of sharing the same laboratory conditions since many generations, continue to harbor a signicant regression with the temperatures of their origins (R2 = 0.889, P = 0.016). From Jaramillo, N., PhD thesis.
convergence at the same laboratory temperature, and its association with temperature of initial eld conditions, suggest a genetic determinism for size variation. This latter example illustrates that Bergmann size clines in natural populations may have more complex causes than a merely developmental process (David et al., 1994; Vanvoorhies, 1996). Shape, as dened by geometric morphometrics, also shows evidence for strong genetic determinism. Actually, shape appears as a classical polygenic character (Klingenberg and Leamy, 2001; Breuker et al., 2006a; Patterson and Klingenberg, 2007). When studies on quantitative trait loci (QTL) were applied to the shape and size of mouse mandible, many QTL were identied for shape (Klingenberg et al., 2004), many more than for size (Workman et al., 2002; Klingenberg et al., 2001). Fewer studies are found in insects, also tting the idea of genetic determinism (Iriarte et al., 2003) and polygenic inheritance (Shrimpton and Robertson, 1988; Long et al., 1995). 2.1.4. Genetic assimilation, genetic accommodation In Andersons experiments and in Jara-millos observations, genetic differences for size acquired by genetic assimilation have been suggested. Waddington (1953) dened genetic assimilation as a process by which a phenotypic character, which initially is produced only in response to some environmental inuence, becomes, through a process of selection, taken over by the genotype, so that it is found even in the absence of the environmental inuence which had at rst been necessary (Waddington, 1953). Meanwhile, Schmalhausen had given this concept the name of stabilizing selection (Gilbert, 2000; Levit et al., 2006). Both scientists used orthodox genetics to explain phenomena that had been (wrongly) considered cases of Lamarckian inheritance of acquired characteristics. Genetic assimilation is the genetic xation of one of two or more phenotypes that had been adaptively expressed. Through this concept phenotypic plasticity acquires an evolutionary importance (Braendle and Flatt, 2006). Genetic assimilation has been extended to the genetic accommodation concept (West-Eberhard, 2003), and received some experimental support (Waddington, 1956; Rutherford and Lindquist, 1998; Sollars et al., 2002; Suzuki and Nijhout, 2006). The model of genetic accommodation assumes that the trigger uncovering previously cryptic genetic variation is either genetic or environmental, whereas the concept of genetic assimilation typically assumes only an environmental trigger. Thus, genetic accom-
880
2007). Accordingly, geometric shape was also intermediate between parents (Dujardin, unpublished data). These observations are in agreement with allopatric conspecic T. protracta populations instead of with subspecies. Indeed, genetically differentiated parents, as different subspecies or distinct (but phylogenetically close) species, may produce an exaltation of some body dimensions and other phenotypic traits in their progeny. Centroid size was increased in hybrids obtained from two close species, initially considered as two subspecies (Costa and Felix, 2007), Triatoma brasiliensis and Triatoma juazeirensis. It was larger than the midparent size, and larger than the largest parents size, suggesting heterosis pointing to a consistent genetic divergence of the parents. Extending this study to experimental hybrids among the four members of the Brasiliensis complex, it was possible to show a linear relationship between the genetic divergence of the parents (Costa et al., 2001; Costa and Felix, 2007) and the increase in size of their offspring (Costa et al., pers. comm.). 2.2.3. Mutation Mutations are evolutionarily important but quite infrequent events. In medical entomology, mutations conferring resistance to insecticides or adaptation to a new environment are highlighted. Unless the phenotype would be directly modied by the mutation, morphometrics does not detect such changes. However, mutations may induce a morphometrically detectable tness cost in the previous environment (Bourguet et al., 2004). Resistance ace-1 alleles coding for a modied AChE1 in Culex, as well as Ester 1 and Ester 4 resistance alleles, were associated with shorter wing length together with other tness related traits, with higher mortality in less developmentally stable specimens (Bourguet et al., 2004). Morphometric techniques analyzing developmental stability could be useful: as explained in the next section, they relate to heritable and non-heritable changes and may provide indirect and exclusive information about the biology of an organism. 2.3. Non-heritable changes One specic talent of morphometrics is its ability to reveal variation attributable to the environment only, i.e. to remove the possible genetic sources of the variation and focus on its environmental origin. Non-heritable variation is then partitioned from measurement error, and understood as an effect of the developmental response to external uctuations. This task is performed by comparing bilateral structures, such as wings, legs, or structures such as both sides of the head (matching symmetry) (Klingenberg et al., 2002). Since these are structures belonging to the same genotype, environmental causes remain the most likely source of asymmetry. Things are not that simple however, since asymmetry might be genetically programmed especially for internal organs or affected by modier genes (genes that have small quantitative effects on the level of expression of another gene). Development is made up of a complex series of binary decisions with discrete outcomes governed by thresholds: any mutation or environment that brings an individual closer to such a threshold could increase the sensitivity of that particular developmental outcome to stochastic perturbations (Milton et al., 2003). Only uctuating asymmetry6 (FA) is considered as a non-heritable variation,
generally an environmentally induced one (Van Valen, 1962; Palmer and Strobeck, 1986). FA has endogenous and exogenous causes (Graham et al., 1993). Among the endogenous sources of FA are inbreeding processes (Soule, 1979) and interspecic hybridism. Inbreeding processes would affect developmental stability because of increasing homozygosity, while outbreeding would affect it because of the disrupted coadaptation of two genomes (Graham, 1992; Clarke, 1993) but see Alibert et al. (1994), Largiader et al. (1994). Exogenous causes of developmental instability are generally presented as an environmental stress, such as exposure to sublethal dosis of insecticides during development (McKenzie and Clarke, 1988; Clarke and McKenzie, 1992; Mpho et al., 2000). Insecticides at sublethal doses may increase FA at some characters, but not others (Chang et al., 2007). Thus, departure from symmetry (FA) has been used as a measure of biological problems (for a review, see Moller and Swaddle (1997)). Populations at the limits of the species territory generally show higher levels of FA, because of both extreme environments and possible contact with other species (introgression, hybridism) (Graham et al., 1993; Dujardin, 1998; Schneider et al., 2003). However, the removal of a permanent stress may also be assumed to destabilize development. Domesticity would increase the level of FA not because of stress, but because of the removal of selection pressure against unt phenotypes. This mechanism was suggested to explain why domestic animals tend to be more asymmetrical than wild animals (Parsons, 1990). In Bolivian T. sordida, domesticity was associated with higher FA (Dujardin et al., 1999f). Since these are non-heritable changes, their use in medical entomology makes sense where it is relevant to question the environmental effects on the organism, or where it is possible to use them for identifying individuals (see Section 5.1) or more generally for characterizing populations. Comparing variance of individual traits also may provide information on environmental inuence, although it might be also an indication of genetic effects. In R. prolixus, the variance of centroid size in domestic habitats was twice as large as the one observed in sylvatic conditions (palm trees), where smallest and largest individuals were seemingly absent (Feliciangeli et al., 2007). Very few studies have been conducted in this eld (Houle, 1992; Mller, 1992; Pomiankowski, 1997). 3. From dimensions to epidemiology In medical entomology, the objective is to reduce or eliminate the populations of insects transmitting diseases. This objective may require heavy economical investments, so that there is a need for ne-tuning the target. To conrm the target, an accurate species diagnostic must be performed. To apply control methods, a mapping of target populations must be set up. Moreover, medical entomologists need to know the way each vector organizes its populations: into separate subpopulations, or into connected groups. This knowledge can improve control strategies before the phase of attack, and it can support entomological surveillance after the campaigns. 3.1. Target identication 3.1.1. The target as a species For each vector, there is generally a relatively stable territory where the species can be found, and beyond which it is absent or found only exceptionally. The geographic extension is an important trait of the vector, especially for making predictions about its epidemiological relevance or the costs of a control programme. Reliable diagnostic tools are necessary to assess the geographic
6 Fluctuating asymmetry (FA) must be distinguished from directional asymmetry (DA) and antisymmetry (AS), both (DA, AS) commonly thought as of genetic origin. The distinction between these asymmetries is based on the average of bilateral signed differences (larger than zero if DA), the distribution of the signed differences, either a normal one (FA) or not, and the magnitude of the bilateral differences, either a constant one (AS) or not.
881
extension of a given species (see Section 4.1.2). Furthermore, although various distinct species may transmit the same pathogens, there is often a reduced set of them actively transmitting pathogens. As long as they harbor different morphological features, their determination may rely on traditional techniques of entomology. However, many vectors are forming groups of morphologically close species, some of them morphologically indistinguishable from others, vectors or not. These biological species have received names suggestive of this lack of morphological distinction: cryptic species, sibling species and isomorphic species. As a consequence, one of the most important needs in medical entomology has been an identication tool able to accurately separate species however close they may be morphologically. Thus far, in the absence of reliable qualitative, morphological trait allowing discrimination, this tool has been derived from genetic techniques. For these morphologically close species and for others, morphometrics has an important role to play (see Section 4.1). 3.1.2. The target as a local population Within the species recognized as vectors, the target may be further restricted to subpopulations because they present a higher risk of transmission, they transmit different pathogens, they present different levels of vulnerability, or they are isolated subpopulations. Adaptation to human habitat is often the epidemiologically relevant mechanism of subpopulation constitution. When coming in close association with humans, the vectors are more likely to transmit the pathogen to humans, increasing the prevalence of the disease (Campbell-Lendrum et al., 2001), and the vector adapts to new conditions of life (Tapis and Hausermann, 1975, 1978). As described for T. infestans (Dujardin, 1998), which can probably be extended to other vectors, successive steps in adaptation to humans add their effects to build vector subpopulations: (i) Since only part of the sylvatic genotypes can be successful in establishing sustainable domestic colonies, some restriction of genetic variability is assumed during the early domestication process. (ii) The insects dispersion may be enhanced by its dependence on its host; thus the passive spread of adapted insects occurs. The insect is likely to be transported by humans over large geographic distances, out of the current range of its ecological constraints. (iii) As a consequence of both isolation from original sylvatic foci and founder effects in the new areas of colonization, a further change in genetic properties is expected. (iv) As long as this geographic expansion goes on, domesticity becomes a more exclusive habit, and some populations with high levels of inbreeding may show external evidence of developmental instability, such as increased uctuating asymmetry or unilateral morphological monstrosities (Graham et al., 1993; Dujardin, 1998; Dujardin et al., 1999f). It seems that morphometrics can provide help in identifying these subspecic targets, because signicant intraspecic variation is commonly found, frequently related to geography. For instance, using simple measurements of head dimensions, it has been shown that T. infestans, the main vector of Chagas disease in the Southern Cone countries, and R. prolixus, the most important one in the Northern countries of South America and Central America, present a cline of decreasing size from their supposed geographic origin to the more peripheral areas of their territorial expansion (Dujardin et al., 1998a). Examples of epidemiologically relevant subpopulations are found in all medically important insects. We provide hereafter a
few examples among those which were submitted to morphometric studies. 3.1.2.1. Triatominae, Chagas disease. The vectors of Chagas disease offer a good example of subspecic targets, since from the few species actively transmitting the parasite, mainly their domestic fraction have public health importance. Most Triatominae are potentially able to allow the complete cycle of the parasite, Trypanosoma cruzi (Dujardin and Schoeld, 2004). As such they are potential vectors, but not necessarily active vectors to humans; to become a public health problem, an additional step is needed: their close association with humans (Costa, 1999; Matias et al., 2002). The target then is not the species, but the domestic populations of the species (Schoeld and Dias, 1991). The reason is not only that the latter are a risk for humans, but rather that they are more vulnerable: the cleaning of houses and peridomestic dependencies with insecticide products is possible, whereas this task is inconceivable in sylvatic foci (Schoeld and Dias, 1998; Dujardin and Schoeld, 2007). 3.1.2.2. Ae. aegypti, dengue and yellow fever. Ae. aegypti is another example where the target is redened within species. This mosquito is the vector of yellow fever in some important parts of the world; and of dengue worldwide. Actually the vector to humans is the domestic fraction of the species, i.e. the one in close association with humans (Edman, 2003). Sylvatic Ae. aegypti are not a feasible target nor are they a direct risk for humans (Leahy et al., 1978; Beard et al., 1985). Moreover, within the domestic populations of South East Asia, various techniques have conrmed the existence of more or less differentiated subpopulations of Ae. aegypti. Mark-release recapture experiments in Puerto Rico and Thailand demonstrated that adult Ae. aegypti dispersed relatively short distances (Harrington et al., 2005). In apparent contrast with these observations, the level of isozymic differentiation in Thailand was low (Sukonthabirhom et al., 2005). A further study based on mtDNA of Ae. aegypti suggested that urban areas of Thailand were relatively panmictic, while suburban/rural sites exhibited more restricted gene ow (Bosio et al., 2005). The geometry of the wing of Ae. aegypti showed signicant differentiation among large geographic areas of Thailand (Jirakanjanakit and Dujardin, 2005), but was obtained from relatively old laboratory lines. Unpublished data on eld-collected specimens revealed less, but still signicant structuring in global accordance with geography (Jirakanjanakit et al., unpublished data). 3.1.2.3. Phlebotominae and Leishmania sp.. Sandies raise many unsolved questions for the taxonomist, especially at the generic and higher levels, while at the species level, it is not uncommon to detect cryptic species (Dujardin et al., 1996b; Ward et al., 1988). They present highly differentiated geographic or ecologic subpopulations, raising frequent doubts about their conspecicity, so that the few morphometric studies conducted on geographic subpopulations often questioned their species status also (Dujardin et al., 1997c, 1999d, 2005; De la Riva et al., 2001; Dujardin and Le Pont, 2000). Lutzomyia nuneztovari (Diptera: Phlebotominae) is a striking example of a single species transmitting two parasites in close geographic areas (Torrez et al., 1999; Martinez et al., 1999). This situation has been interpreted as a consequence of the increasing domesticity of Lu. nuneztovari in the Yungas of Bolivia (Campbell-Lendrum et al., 2001). Transmission of different pathogens by the same species could be a side effect of domesticity, but we cannot exclude subpopulations of Lu. nuneztovari having different parasite specicities. In this species as well as in many other medically important sandies, subpopulations are fre-
882
quently observed with either different vectorial capacities, different domestic adaptations, or presenting possible speciation processes (Lane and Marshall, 1981). 3.1.2.4. Africa, or the vectors of sleeping sickness. Two studies have been conducted on local populations of Glossina palpalis gambiensis, the vector of sleeping disease in West Africa, using both genetic and morphometric techniques on the same specimens (Camara et al., 2006; Bouyer et al., 2007). While separated subpopulations were conrmed by both techniques between island and continental populations of tsetse ies (Camara et al., 2006), very low levels of separation could be detected elsewhere (Bouyer et al., 2007). In both studies, there was a general agreement between genetic and metric variation among geographic locations. 3.2. Entomological surveillancePopulation structure Frequently, target populations are geographically localized ones. But an insect species does not occupy its geographic territory in a uniform manner, it organizes in populations according to its reproductive strategy, its dispersion power, its hosts preferences or dependence, and its local adaptation capacities (Dujardin, 1998; Schoeld et al., 1999). With evolutionary time frames, such subpopulations were trapped by natural events such as orogenesis, glaciation, inundations, etc. and became separate entities. Without considering the evolutionary time scale, some current subpopulations of a vector diverged because of both genetic drift and local selection pressure. Specialized tools are needed to know if they are connected by frequent migrants, or if they are isolated subpopulations. A valid control strategy may depend on the answer to this question. In case of high structuring such as that found in T. infestans (Dujardin et al., 1998b), the main vector of Chagas disease, a sequential strategy can be applied without a signicant risk of reinfestation of one area by another. When this structuring is not evidenced, where exchanges are suspected among the vectors domestic and sylvatic foci such as in R. prolixus in Venezuela (Harry, 1992; Feliciangeli et al., 2007; Fitzpatrick et al., 2008), T. brasiliensis (Borges et al., 2000) or T. dimidiata (Dorn et al., 2003), all of them are important vectors of Chagas disease (Dujardin et al., 2002), innovative strategies have to be implemented. To characterize these subpopulations, to understand their evolutionary or microevolutionary meaning, to track them, genetic techniques are used. In complement to them, and sometimes as a valid alternative, morphometric techniques may be a valuable characterizing tool. For instance, in case of reinfestation, the structured pattern of the population makes it possible to apply morphometric techniques to detect the geographic origin of recent reinfestation (Dujardin et al., 2002, 1996a). 4. Dening the target 4.1. Species The recognition of species, even cryptic species, is generally possible using morphometric techniques (Lane and Ready, 1985). Morphometrics has been used to detect new species (Dujardin et al., 1999c,d, 2005), and to contribute to their complete description (Le Pont et al., 1997a,b). The taxonomic power of morphometrics has also been used to question species boundaries (Dujardin et al., 2004), or to synonymize controversial taxa (Gumiel et al., 2003). Cryptic species, when examined for morphometric differences, have been accurately distinguished in Triatominae (Villegas et al., 2002), sandies (Lane and Ready, 1985; De la Riva et al., 2001.) and mosquitoes (Kitthawee et al., pers. comm.).
4.1.1. Species detection What is the reliability of species detection using characters inuenced by the environment? Is metric variation observed among conspecic populations (Lane and Marshall, 1981; Hutcheson et al., 1995; Dujardin and Le Pont, 2004) able to interfere with interspecic variation (Le Sueur and Sharp, 1991; Dujardin et al., 2003)? Although there is no special metric feature marking the difference between species, this topic is inuenced by common ideas found in the literature, neither completely true nor false. 4.1.1.1. The amount of metric difference. The level of interspecic difference is generally (much) higher than the corresponding intraspecic variation, even across geographic populations. This led some authors to look for how much species differ in general (Thackeray et al., 1997), but this would need to be addressed separately for each group (Dujardin et al., 2003). 4.1.1.2. The anatomical localization of metric differences. When metric differences are found in organs responsible for mechanical reproductive barriers, they may be given more evolutionary importance than differences located elsewhere. It is well-known that sandy species generally have different dimensions or shape in at least one piece of their complex genitalia (Young and Duncan, 1994). The properties of metric differences. It is generally believed that species differ not only in size, but also, and probably more, in shape (Atchley, 1983). When shape difference is allometry-free, it means that more than size differs, which is not common among conspecic groups unless ecological differences are very important (see Section 2.1; Fig. 5). There is, however, no rigorous way to attribute shape differences, even allometry-free shape differences, to the speciation process instead of to locally adaptive causes not involved in speciation (Schilthuizen, 2000). The spatial distribution of the organisms. Are the individuals able to cross in natural conditions?
Fig. 5. Intraspecic variation recorded for ten species of Neotropical sand ies among areas classied as located within or between the ve major eco-geographic regions of the South American continent. Quantile plots show range of variation in size (top) or size-independent (bottom) divergences between two conspecic populations, according to whether they belong to the same eco-geographic region (W for within region) or to distinct ones (B for between regions). Size was the rst common principal component and shape was the set of remaining principal components. Size variation was estimated by Pearsons CRL index (1926); shape divergence was estimated by the Mahalanobis distances. Each box shows the group median as a vertical line across the middle and the quartiles (25th and 75th percentiles) as its ends. The 10th and 90th quantiles are shown as lines on each side of the box.
883
4.1.1.3. Sympatry. When compared groups are sympatric, i.e. they occupy the same area or inhabit the same place, then size or shape differences may suggest assortative mating. Consistent metric differences between groups living in contact with each other suggest reproductive isolation; they are a good indicator, although not a proof, of speciation (Busvine, 1978; Claridge and Gillham, 1992; Dujardin et al., 1999c). 4.1.1.4. Allopatry. When differentiated groups are separate geographic populations, assortative mating can no longer be an argument, and metric differences, if any, may be explained by diverging selection acting on quantitative characters. Signicant information may sometimes be obtained from the relationship of size and temperature. According to Bergmanns rule, variation in the size of conspecic organisms should conform to variation in temperature (see Section 2.1). The failure to show such agreement could be an argument for suspecting a speciation process among allopatric populations (Marcondes et al., 1998; Dujardin et al., 1999d). This argument has been used in organisms other than insects (Brunner et al., 2002). 4.1.2. Species identication The problem with the many transformations of the data from simple distances to multivariate factors, or from raw coordinates to partial warps (displacements of landmark relative to the consensus form) and relative warps (the principal components of partial warps), is that the determination of one individual cannot be based on a direct comparison of measurements. The metric traits of a single individual must be entered into a model intended to be sufciently discriminant among species. This model has to be built from a database gathering various species, with a representative number of individuals and geographic origins. 4.1.2.1. Metric key for species identication. The use of quantitative, metric traits to identify species cannot and should not follow the model of dichotomous keys in use with qualitative, morphological characters. However, it should reach an acceptable level of simplicity in order to be integrated into the routine entomological surveillance. The metric key to identifying species would consist of a database and a related model of individual classication. The database should be either a single source web page or a network of electronic databases, and could benet from the rapid progress in information technology, as well as from some already running examples in computer assisted taxonomy (Platnick et al., 2005; Scoble et al., 2007). It would contain mainly those species for which molecular tools are presently needed for accurate diagnostic. The related model of classication should be able to compare individuals characterized by many variables. The statistical tool commonly used to separate at best various groups or species on the basis of multivariate characterization is (multivariate) discriminant analysis (or canonical variate analysis) (Albrecht, 1980). It can be regarded as a MANOVA (multivariate analysis of variance) collapsed to a single classication design (Rohlf et al., 1996). This analysis provides a set of discriminant factors (the model) which can be used to localize a single individual on a factorial map, or to assign it to a given group by measuring distances between it and the different groups (generalized or Mahalanobis distances). The analysis is quite robust, unless sample sizes are very unequal (Pimentel, 1992). Other clustering models exist which are less commonly used in modern morphometrics, such as the K-Nearest Neighbor (KNN) classication or the articial neural network (ANN). In the KNN approach, an individual is assigned to the class most common amongst its K nearest neighbors: the method is simple but requires the selection of an optimal number of neighbors, a task
which can be performed by machine learning techniques (Duda and Hart, 1973; Agrawala, 1977). The ANN approach is a more complex algorithm; it is a slightly less efcient alternative than the discriminant model, but it is more robust against sample dependencies, it applies to any kind of multivariate data sets (Fedor et al., 2008) and is amenable to complete automation (MacLeod et al., 2007). In arthropod classication problems it has been used for pattern recognition on transformed digital images (Do et al., 1999) or on morphometric data (Marcondes and Borges, 2000; Baylac et al., 2003; Vanhara et al., 2007). 4.1.2.2. Database. Ideally, the database would contain the coordinates of several landmarks for many individuals of different species. The discriminant analysis would then need as input the coordinates (after Procrustes superimposition and their transformation into partial warps, see Section 1) of a relevant set of species likely to contain the unknown individual. There is, however, an important problem in making that vision a reality: the measurement error (Conner and Shenk, 2003). The measurement error, which is unevenly distributed among different shape variables (Jordaens et al., 2002; Arnqvist and Martensson, 1998), exists at various steps of morphometric analysis (Arnqvist and Martensson, 1998). Its impact can be reduced in a number of ways; and generally requires averaging repeated collections of the data (Arnqvist and Martensson, 1998). Even after such correction, measurement error may become a signicant obstacle for different users (Jordaens et al., 2002; Rasmussen et al., 2001). As a consequence, user A should not enter his own measurements in a database of coordinates collected by user B, and vice versa. The solution to the multiple users problem is simple but demanding in terms of electronic memory and user effort: instead of coordinates, the database must contain the digital pictures from which coordinates can be collected. Then any user could achieve the complete set of analyses with his or her own digitization data. 4.1.3. Interspecic relationships Comparing morphometric properties among species can help to classify them according to their general similarities as estimated by metric distances. From such data one can build a phenetic tree (a phenogram). Since metric properties are not only the product of environmental effects, but also the reection of the insects genetic make up, this phenetic tree may provide some parallelism with evolutionary trees (Dujardin et al., 1999b,e; Noireau et al., 2002; Costa et al., pers. comm.). Attempts have been made to improve the phylogenetic signal perceptible in the metric classication trees, by coding the continuous data into new, discrete variables, or by relying on various properties of ordination methods. 4.1.3.1. Coding. In geometric morphometrics, the propositions were oriented to the transformation of shape variables into discrete characters. Morphological or molecular characters are discrete data, having two states (present or absent), allowing probabilistic models to be built, and most phylogenetic software has been developed for such discrete variation. The attempts to adapt geometric shape to phylogenetic reconstruction would previously have transformed them into discontinuous characters (Zelditch et al., 1995; Fink and Zelditch, 1995; Swiderski et al., 2002). This type of transformation certainly removes information, and for geometric shape does not prevent a number of theoretical problems (Rohlf, 1998; Adams and Rosenberg, 1998). These were circumvented by an original approach where discrete characters are produced by discontinuities in trait variance (MacLeod, 2002). This approach still suffers from the loss of shape information (Adams et al., 2004).
884
4.1.3.2. Without coding. Shape variables may also be used without converting them into integer codes (Adams et al., 2004). Instead of partial warps (displacements of landmarks relative to the consensus form), it would be more appropriate to use their canonical variates because they are uncorrelated characters (by construction) and they focus on unshared variance (Pimentel, 1992), an attribute which has been likened to an apomorphic character (Sorensen, 1992; Foottit and Sorensen, 1992). Provided an outgroup is included, methods based on continuous variation (continuous maximum-likelihood) or distances based methods (Farris, 1972) (neighbor joining method) could then be used to tentatively portray phylogenetic relationships. 4.2. Individual recognition To assign single individuals to species or subspecies is typically what morphometrics should be able to do as a systematic tool. As suggested above (see Section 4.1.2), the metric key has nothing to do with the classical taxonomic or dichotomous key, and is actually made of a multivariate model by which an individual is projected onto a relevant set of species. As far as we know, this idea has not been applied routinely in the context of entomological surveillance, but a few related examples suggest that this type of tool would perform adequately. Indeed, it has been applied successfully to recognize individuals belonging to less separate entities than species: subspecies, local populations and laboratory lines. Among laboratory lines of Ae. aegypti from different geographic origins in Thailand, the individual classication produced correct assignment in more than 90% of the assays (Jirakanjanakit and Dujardin, 2005). When testing for reclassication of individuals of T. protracta from one generation to the previous one, a single wing, male or female, was assigned to its close relatives in more than 90% of cases (Dujardin et al., 2007). The same method was applied again to Ae. aegypti, going up to ten generations back using data published in Jirakanjanakit et al. (2008): after ten generations, each wing was assigned to its remote parents with the same rate of correct attributions (80%) as after one generation. These data suggest that well discriminated groups remain well discriminated even after ten generations; of course, poorly discriminated groups would also remain poorly discriminated at the next generation. They give condence that among species this method would give satisfactory results. 5. Natural population structuring 5.1. Genetic structure and population structure An important need in medical entomology is to quantify the current exchanges of individuals among subpopulations. This quantication would inform on population structure, to be distinguished from genetic structure which is dened by the level of gene ow among subpopulations. Although mark-recapture studies might be a valid option to evaluate the frequency of migrants among subpopulations (Tapis and Hausermann, 1975; Harrington et al., 2005), it cannot account for passive migration of non-ying stages of the insect, so that this frequency is currently approximated by the measurement of gene ow (Slatkin, 1985, 1981). 5.1.1. Gene ow and the ow of migrants Gene ow measurement provides information on the level of migration among subpopulations. However, this information is of unequal value depending on its output, either lack of gene ow, or complete gene ow. Lack of gene ow is supposedly valid information since in that circumstance (genetic divergence)
migrants are highly unlikely. However, in case of complete gene ow, the epidemiological question is to know whether this genetic structure is a reection of the current level of migration (see the Ae. aegypti example, Section 3.1.2). How contemporaneous or recent it depends on the effective size of the populations under study and the evolutionary rate of the genetic marker (McKay and Latta, 2002). Additional problems with genetic markers are that they are relatively costly and they need appropriate infrastructures. As an unfortunate consequence, genetic markers often remain inside research laboratories and have not yet found their way into routine medical entomology. Modern morphometrics is tempting as a candidate population marker because it is a fast, lowcost, easily spread tool, it is informative about current or very recent population events, and it contains information on genetic variation. However, as long as morphometric traits have much higher environmental variance than genetic markers, they are not appropriate for gene ow estimation. As a consequence, they are not appropriate for estimating the level of migrants. Due to their sensitivity to diversifying selection among subpopulations or habitats, they could underestimate true levels of migration. Metric structuring of natural populations reects both adaptation to local conditions and possible lack of migration, without reliable means to separate these two causes. There are, however, good reasons to assess metric structuring of natural populations. First, this structuring, when it is strong, provides condent results in reinfestation studies (see Section 5.1). Second, when related to temperature variation, it helps in species or speciation process detection among allopatric populations (see Section 4). Finally, since phenotypic plasticity has been shown to have evolutionary importance through epigenetics and genetic accommodation (see Section 2.1), it improves our knowledge on the evolution of the species. 5.1.1.1. Fst and Qst. The importance of diversifying selection inating metric structuring of natural populations can be estimated by comparing on the same material the measurement of Fst as derived from neutral molecular markers and Qst as computed from metric variation. Qst separates quantitative genetic variation in a manner analogous to Fst for single gene markers (Spitze, 1993): if the quantitative characters and the molecular characters are neutral, Qst and Fst should converge to the same value (Hiernaux, 1977; Rogers and Harpending, 1983; Whitlock, 1999). Very few data exist comparing molecular (Fst) and quantitative traits (Qst) measurements of a population structure. They tend to show the following trends: (i) Qst is generally higher, or much higher, than Fst,7 and (ii) the value of Qst depends on character tness (McKay and Latta, 2002). Within species, traits experiencing the strongest local selection pressures are expected to be the most divergent from molecular Fst (McKay and Latta, 2002). The small set of comparisons reported here in medically important insects between (unpublished) Qst and (published) Fst (Table 1) conrm the importance of selection modifying the geometric variation among subpopulations (Fig. 6). It also highlights the higher sensitivity of size (relative to shape) in response to diversifying selection (Fig. 6), which again is in agreement with the idea of shape having less environmental variance than size (see Section 2.1; Fig. 5). 5.1.1.2. Population structure and morphometrics: reinfestation. Metric characters can neither be used directly to compute gene ow, nor can they be used to accurately quantify the exchange of individuals among subpopulations. Nevertheless, as long as they
7 Infrequently, Qst is lower than Fst, suggesting homogenizing selection acting on the quantitative trait.
885
Table 1 Estimations of Qst and standard deviation (stdev) for the average of all the relative warps (all RW), which are the principal components of the partial warps (shape variables), and for the centroid size (CS). Species, Locality Sex Qst (stdev) all RW Qst (stdev) CS 0.92 (0.03) 0.45 (0.01) 0.72 (0.02) Fst 0.055 0.057 Ref.
Aedes aegypti, Bangkok area, Thailand F 0.45 (0.02) Glossina palpalis gambiensis, Loos Island, Guinea F 0.35 (0.04) M 0.32 (0.04) Glossina palpalis gambiensis, Mouhoun river, Burkina Faso F 0.51 (0.03) M 0.44 (0.03) Glossina palpalis gambiensis, Mouhoun and Comoe rivers, Burkina Faso F 0.31 (0.05) Rhodnius prolixus, State of Barinas, Venezuela F 0.57 (0.03) M 0.58 (0.03)
(a) (b)
0.77 (0.06) 0.68 (0.06) 0.45 (0.02)
0.012
(c)
0.008
(d)
0.71 (0.01) 0.35 (0.01)
0.080 to 0.040
(e)
The right-hand columns are Fst and literature references (Ref.): (a) (Sukonthabirhom et al., 2005; Jirakanjanakit, 2007) (b) (Camara et al., 2006) (c) (Bouyer et al., 2007) (d) Bouyer et al., submitted (e) (Fitzpatrick et al., 2008). Although Qst is trait specic, it has been averaged over the complete set of RWs, considering them as dening a single trait: the shape of the wing.
are able to identify the parental generation and to distinguish it satisfactorily from other subpopulations (Falconer, 1981; Dujardin et al., 2007), the morphometric characters might be able to provide relevant information to medical entomologists. This has been demonstrated by case studies of reinfestation after treatment in Triatominae, using traditional morphometrics (Dujardin et al., 1997b), or both genetic and traditional morphometric techniques (Dujardin et al., 1999a). Provided that samples were available from the population before insecticide application, relative metric similarities could suggest the origin of reinfesting specimens: they are either the descendants of previously killed bugs, or immigrants from an external focus. Traditional morphometrics (head measurements) of the Chagas disease vector Triatoma infestans provided information that could identify the source of reinfesting specimens (Dujardin et al., 1997b), and such information has been shown to be in agreement with genetic markers (Dujardin et al., 1999a). The geometry of the wing of the North American T. protracta was tested on laboratory populations and was shown to be an interesting candidate to assess the origin of a given individual (Dujardin et al., 2007). Recently, the geometry of the wing of R. prolixus in Venezuela has been used to understand reinfestations of houses and of peridomestic structures after
insecticide application in the Province of Barinas (Feliciangeli et al., 2007). The conclusions of this study have been fully supported by a later genetic study (Fitzpatrick et al., 2008). Since a residual population is assumed to be the same generation as or the next generation to the individuals subjected to insecticide spraying, the reinfestation analysis is based on the assumption that an insect is more similar to its parents than to other insects. But the reasons for successful results are not only the higher similarity among relatives. In the reinfestation studies, the objective is not to measure gene ow or levels of migration, but rather to distinguish local inhabitants from foreigners (so to speak). Thus, the environmental effect on metric traits is a welcome effect. Insects reared in the same microenvironmental conditions (a few houses, a village) would share a signicantly larger amount of metric similarity, making a residual population easier to recognize. Of course, the level of population structuring is an important condition for morphometric characters to be applicable to reinfestation studies. They would be less applicable to highly dispersive insects breaking the population structure at each new generation. 6. Conclusions and perspectives We have presented and commented on what appears to us to be the currently main or advisable applications of morphometrics to medical entomology (see Section 3), namely (i) species identication (ii) species detection, (iii) reinfestation studies and (iv) population structure studies. 6.1. Species identication Traditional morphometrics had already established virtues for species recognition and population characterization. Modern morphometrics adds power and ease to this central application. What is now possible with the development of digital images has not been achieved yet, i.e. a database of pictures of the different medically important species (see Section 4.1.2). Such a database, which could be established by countries, or by the domestic political subdivisions within each country, would give morphometrics its full power for species identication, detection (but see hereunder) and in general for entomological surveillance. No doubt it would help decrease the need for the more costly molecular markers in use today.
Fig. 6. Bars represent the Qst/Fst ratio (see Table 1). Although Qst is trait specic, it has been averaged over the complete set of relative warps, considering them as dening a single trait: the shape of the wing. Shape, the average of all relative warps. Size, centroid size; G.p.g., the tsetse y Glossina palpalis gambiensis; f, females; m, males; B, Bangkok (Thailand); L, Loos Islands, Guinea; M, Mouhoun river, Burkina Faso. C, Comoe river, Burkina Faso (Bouyer et al., unpublished data). The ratio is not shown for R. prolixus from Table 1 because Qst and Fst were not derived from the same specimens.
886
6.2. Species detection This task may be broader than a simple morphometric task since it refers to the biological question of speciation. Dening species as reproductively isolated populations, the detection of xed differences among sympatric specimens is an informative signal, but is not proof of speciation. Detection is even more delicate among allopatric samples. Interestingly here, the impact of the environment on metric variability is helpful, since speciation is expected to break some of the current rules observed among conspecic populations, such as the Bergmanns rule (see Section 4). 6.3. Reinfestation Reinfestation studies in the absence of insecticide resistance is a particular application of population structure studies (see hereunder Section 6.4). The objective is not to estimate the number of migrants between groups, but instead to decide whether the individuals at hand are foreigner or local inhabitants, so to speak. Since the comparisons are within one or two generations, the problem is also to assign individuals to their closest relatives. In this application, both inheritance and plasticity of metric traits help to obtain correct attribution. The important condition to make this application feasible is strong population structuring, whatever its cause, either genetic drift, environmental inuence, or both (see Section 5.1). Thus far, reinfestation studies have been conducted and tested on Chagas disease vectors only. 6.4. Population structuring Since phenotypic plasticity has been shown to have evolutionary importance (see Section 2.1), the metric structuring of natural populations is a biologically relevant trait. But in medical entomology, the most desired information on a given population is its level of isolation from neighboring populations. Some of the most successful control measures have been possible on island populations because islands do not represent a high risk of reinfestation (see Schoeld and Patterson (2005)). Continental populations are sometimes also isolated from neighboring ones, representing biogeographical islands where control measures can be taken without risk of quick reinfestation. Is the morphometric marker able to detect biogeographical islands? The answer is yes, with however the drawback of a possibly high rate of false positives. Among isolated populations, both genetic drift and environmental differences act together to produce metric differentiation, often with a consistent shape differentiation not attributable to size variation. However, because of the inating effect of local selection on metric variation, connected subpopulations could show consistent differences as well. As suggested by the generally Qst > Fst (see Fig. 6), the reverse is not expected as a common feature, i.e. a lack of metric differentiation between isolated populations. Thus, by simply discarding the likely connected subpopulations, the ones lacking metric differentiation, morphometrics can be used as a preliminary exploration tool to identify putative biogeographical islands. Among them, the ones harboring strong allometry-free divergence should be considered as good candidates. They would then be the subject of analyses based on genetic markers and/or mark-release recapture techniques. In order to estimate the possible false positive rate, more studies are needed comparing gene ow as measured by Fst and metric structuring as estimated by Qst. 6.5. Recent perspectives Modern morphometrics is itself a quickly evolving discipline, and the medical entomologist should observe its current devel-
opment in both data collection and statistical analyses. New software is being developed to automatize tasks (Tolski, 2004), progressively reducing the human artifact (the user effect, see Section 4.1.2) in data collection. An emerging application of morphometrics is related to evolutionary developmental biology, a discipline (evo-devo) focusing on the role of development in adaptive evolution (Breuker et al., 2006b). As suggested from some examples reported here (Section 2.1), it could provide new tools to improve our understanding of the biology of medically important insects. 7. Authors software for geometric morphometrics Most specialized and generally free software products are listed and available at http://life.bio.-sunysb.edu/morph. The author has his own page at http://www.mpl.ird.fr/morphometrics, also reported on the rst web site. All the software is free (General Public License) and downloadable from the web site. There are various modules, one for collecting data (COO), one for modifying the data (TET), one for Procrustes superimposition and visualization (MOG) and the others for analyzing coordinates (PAD, COV, ASI). 7.1. COO COO (Collection of Coordinates), multiplatform (Windows, Linux) and beta version, is designed to collect coordinates of anatomical landmarks (Fig. 1). It saves the data in the folder where digitized images are stored. This ASCII le has an automatically generated name composed of the word coord_ and the date. It ends by the extension .txt. The internal format is close to the Rohlfs TPS format (see the TPS software at http://life.bio.sunysb.edu/morph). 7.2. TET TET (for the Spanish words Tabla, Espacios, Texto). This module opens and transforms the le generated by COO (coord_data.txt) into two separate ASCII les. The rst one, ending with _FORMAT.txt (not .tps or .nts), contains the coordinates in the format required by the following modules MOG, PAD, COV, ASI (see hereunder). The second one contains these coordinates ready to be copied and pasted into a spreadsheet or database with the additional indication of the internal hard disk path to the picture les, the name of each image, its scale (pixels versus mm), and other users indications. This second le ends with _DB.txt. In addition, TET may perform conversions between some formats, or simple table modications such as row or column removal, concatenation, etc. 7.3. MOG MOG (for the Spanish words Morfometria Geometrica), multiplatform (Windows, Linux) and beta version, allows users to visualize the different steps of a Procrustes superimposition (Fig. 3). It generates ASCII les containing residual coordinates (_ALIGNED.txt), partial warps (or shape variables _PW.txt), centroid sizes (_CS.txt) and a le containing both shape and size (_PW_CS.txt). It offers the possibility to classify unknown individuals on the basis of Procrustes distances using a simplied KNN algorithm (see Section 4.1.2). The output format is ready to use for the multivariate analyses programs described hereunder. 7.4. COV COV (for Covariance), multiplatform, beta version, performs various basic or specialized multivariate operations. Among others,
887
it offers the following tests: principal component analysis (PCA), partial warps (PW), different versus common slope allometric model (MANCOVA), two-state multivariate phenotypic changes (MANOVA, TSMPC), metric disparity (MD), partial metric disparity, Qst, 2 Block Partial Least Squares (2BPLS), angle between hyperplanes, Hotellings T2, Euclidean and Procrustes distances. COV is internally connected to external software allowing more possibilities (see http://www.mpl.ird.fr/morphometrics) 7.5. PAD PAD (for the Spanish words Permutaciones, Analisis Discriminante), multiplatform, beta version, performs a multivariate discriminant analysis (DA) with limited graphical display. PAD allows random permutation of individuals among groups to test the signicance of each pair-wise Mahalanobis distance. It reclassies individuals, with and without cross-validation, and supplementary data can be introduced. For PW data, if requested, PAD rst performs a PCA, then suggests a representative and acceptable set of RW (principal components) as input for the discriminant analysis. 7.6. ASI ASI (for the Spanish word Asimetra) performs a two-way ANOVA for testing asymmetry of size and a Procrustes ANOVA for asymmetry of shape. Acknowledgments I thank Dr. N. Jaramillo (Medellin University, Colombia) and Dr. N. Jirakanjanakit (Mahidol University, Bangkok) for providing unpublished data, and Dr. A. Henry for useful discussion. This work beneted from international collaboration through the ECLAT and LTTRN networks. References
Adams, D., Rosenberg, M.S., 1998. Partial warps, phylogeny, and ontogeny: a comment on nk and zelditch (1995). Systematic Biology 47, 168173. Adams, D.C., Rohlf, F.J., Slice, D.E., 2004. Geometric morphometrics: ten years of progress following the revolution. Italian Journal of Zoology 71, 516. Agrawala, A.K., 1977. Machine Recognition of Patterns. IEEE Press, New York. Albrecht, G.H., 1980. Multivariate analysis of the study of form with special reference to canonical variate analysis. American Zoology 20, 679693. Albrecht, G.H., Gelvin, B.R., Hartman, S.E., 1993. Ratios as a size adjustment in morphometrics. American Journal of Physical Anthropology 4 (91), 441468. Alibert, P., Renaud, S., Dod, B., Bonhomme, F., Auffray, J.-C., 1994. Fluctuating asymmetry in the Mus musculus hybrid zone: a heterotic effect in disrupted co-adapted genomes. Proceedings: Biological Sciences 258 (1351), 5359. Ameneshewa, B., Service, M.W., 1996. The relationship between female body size and survival rate of the malaria vector Anopheles arabiensis in Ethiopia. Medical Veterinary Entomology 10, 170172. Anderson, W., 1973. Genetic divergence in body size among experimental populations of Drosophila pseudoobscura kept at different temperatures. Evolution 2 (27), 278284. Arnqvist, G., Martensson, T., 1998. Measurement error in geometric morphometrics: empirical strategies to assess and reduce its impact on measure of shape. Acta Zoologica Academiae Scientarum Hungaricae 44 (12), 7396. Atchley, W.R., 1983. Numerical taxonomy. In: Some Genetic Aspects of Morphometric Variation, Springer-Verlag, Berlin. Atkinson, D., 1994. Temperature and organism sizea biological law for ectotherms? Advanced Ecological Research 25, 158. Baylac, M., Daufresne, T., 1996. Wing venation variability in Monarthropalpus buxi (Diptera, Ce-cidomyiidae) and the quaternary coevolution of Box (Buxus sempervirens l.) and its midge. A geometrical morphometric analysis. In: Marcus, L.F., Corti, M., Loy, A., Naylor, G.J.P., Slice, D. (Eds.), Advances in Morphometrics. Proceedings of the 1993 NATO-ASI on Morphometrics. Plenum Publ. NATO ASI, ser. A, Life Sciences, New York, pp. 285301. Baylac, M., Villemant, C., Simbolotti, G., 2003. Combining geometric morphometrics with pattern recognition for the investigation of species complexes. Biological Journal of the Linnean Society 80 (1), 8998.
Beard, C.B., Kloter, K.O., Carroll, M.K., Magnuson, L.J., Trapido, H., 1985. Response of domestic and peridomestic strains of Aedes aegypti (Diptera: Culicidae) in New Orleans, Louisiana, USA, to organophosphate, organochlorine, and pyrethroid insecticides. Journal of Medical Entomology 22 (3), 276280. Bergman, A., Siegal, M.L., 2003. Evolutionary capacitance as a general feature of complex gene networks. Nature 424 (6948), 501504. Bird, A., 2007. Perceptions of epigenetics. Nature 447, 396398. Blackmore, M.S., Lord, C.C., 2000. The relationship between size and fecundity in Aedes albopictus. Journal of Vector Ecology 212217 . Borges, E.C., Dujardin, J.P., Schoeld, C.J., Romanha, A.J., Diotaiuti, L., 2000. Genetic variability of Triatoma brasiliensis (Hemiptera: Reduviidae) populations. Journal of Medical Entomology 37 (6), 872877. Bosio, C.F., Harrington, L.C., Jones, J.W., Sithiprasasna, R., Norris, D., Scott, T.W., 2005. Genetic structure of Aedes aegypti populations in Thailand using mitochondrial DNA. American Journal of Tropical Medicine and Hygiene 72 (4), 434442. Bourguet, D., Guillemaud, C., Chevillon, T., Raymond, M., 2004. Fitness costs of insecticide resistance in natural breeding sites of the mosquito Culex pipiens. Evolution 58 (1), 128135. Bouyer, J., Ravel, S., Dujardin, J., de Meeus, T., Vial, L., Thevenon, S., Guerrini, L., Sidibe, I., Solano, P., 2007. Population structuring of Glossina palpalis gambiensis (Diptera: Glossinidae) according to landscape fragmentation in the Mouhoun river, Burkina Faso. Journal of Medical Entomology 44 (5), 788795. Braendle, C., Flatt, T., 2006. A role for genetic accommodation in evolution? BioEssays 28, 868873. Breuker, C., Patterson, J., Klingenberg, C., 2006a. A single basis for developmental buffering of Drosophila wing shape. PLoS ONE 1 (1), e7. Breuker, C.J., Debat, V., Klingenberg, C.J., 2006b. Functional Evo-Devo. Trends in Ecology and Evolution 21 (9). Brunner, S., Shaughnessy, P.D., Bryden, M.M., 2002. Geographic variation in skull characters of fur seals and sea lions (family Otariidae). Australian Journal of Zoology 50 (4), 415438. Burnaby, T.P., 1966. Growth-invariant discriminant functions and generalized distances. Biometrics 22, 96110. Busvine, J.R., 1978. Evidence from double infestation for the specic status of human head lice and body lice (Anoplura). Systematic Entomology 3, 18. Camara, M., Caro-Riano, H., Ravel, S., Dujardin, J., Hervouet, J.P., de Meeus, T., Kagbadouno, M.S., Bouyer, J., Solano, P., 2006. Genetic and morphometric evidence for isolation of a tsetse (Diptera: Glossinidae) population (Loos islands, Guinea). Journal of Medical Entomology 43 (5), 853860. Campbell-Lendrum, D., Dujardin, J.P., Martinez, E., Feliciangeli, M.D., Perez, L.E., Silans, L.N., Desjeux, P., 2001. Domestic and peridomestic transmission of American cutaneous leishmaniasis: changing epidemiological patterns present new control opportunities. Memorias do Instituto Oswaldo Cruz 96 (2), 159 162. Chang, X., Zhai, B., Wang, M., Wang, B., 2007. Relationship between exposure to an insecticide and uctuating asymmetry in a damsely (Odonata, Coenagriidae). Hydrobiologia 586 (1), 213220. Claridge, M.F., Gillham, M.C., 1992. Variation in populations of leafhoppers and planthoppers (Auchenorrhyncha): biotypes and biological species. In: Foottit, R.G., Sorensen, J.T. (Eds.), Ordination in the Study of Morphology, Evolution and Systematics of Insects: Applications and Quantitative Genetic Rationales. Elsevier, New York, pp. 241259, 418 pp. Clarke, G., McKenzie, J., 1992. Coadaptation, developmental stability, and tness of insecticide resistance genotypes in the Australian sheep blowy, Lucilia cuprina: a review. Acta Zoologica Fennica 191, 107110. Clarke, G.M., 1993. The genetic basis of developmental stability. I. Relationships between stability, heterozygosity and genomic coadaptation. Genetica 89, 15 23. Conner, M.M., Shenk, T.M., 2003. Distinguishing Zapus hudsonius preblei from Zapus princeps princeps by using repeated cranial measurements. Journal of Mammalogy 84 (4), 14561463. Costa, J., 1999. The synanthropic process of Chagas disease vectors in Brazil, with special attention to Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae, Triatominae) population, genetical, ecological, and epidemiological aspects. Memorias do Instituto Oswaldo Cruz 94, 239241. Costa, J., Felix, M., 2007. Triatoma juazeirensis sp. nov. from Bahia State, northeastern Brazil (Hemiptera: Reduviidae: Triatominae). Memorias do Instituto Oswaldo Cruz 102, 8790. Costa, J., Monteiro, F., Beard, C.B., 2001. Triatoma brasiliensis Neiva, 1911 the most important Chagas disease vector in BrazilPhylogenetic and population analyzes correlated to epidemiologic importance. American Journal of Tropical Medicine and Hygiene 65 (280). Daly, H.V., 1992. A statistical and empirical evaluation of some morphometric variables of honey bee classication. In: Foottit, R.G., Sorensen, J.T. (Eds.), Ordination in the Study of Morphology, Evolution and Systematics of Insects: Applications and Quantitative Genetic Rationales. Elsevier, New York, pp. 127 156, 418 pp. David, J.R., Moreteau, B., Gauthier, J.P., Petavy, G., Stockel, A., Imasheva, A.G., 1994. Reaction norms of size characters in relation to growth temperature in Drosophila melanogaster: an isofemale lines analysis. Genetics, Selection, Evolution 26, 229251. Davidowitz, G., DAmico, L.J., Nijhout, H.F., 2004. The effects of environmental variation on a mechanism that controls insect body size. Evolutionary Ecology Research 6, 4962.
888
J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890 Dujardin, J.P., Le Pont, F., Baylac, M., 2003. Geographic versus interspecic differentiation of sandies: a landmark data analysis. Bulletin of Entomological Research 93, 8790. Dujardin, J.P., Le Pont, F., Martinez, E., 2004. Is Lutzomyia serrana (Diptera, Pschodidae, Phlebotominae) present in Ecuador? Parasite 11, 211217. Dujardin, J.P., Le Pont, F., Matias, A., De La Riva, J.X., 2005. Morphometrical evidence for speciation within Bolivian Lutzomyia aragaoi (Diptera: Psychodidae). Infection, Genetics and Evolution 5 (4), 362365. Dujardin, J.P., Beard, C.B., Ryckman, R., 2007. The relevance of wing geometry in entomological surveillance of Triatominae, vectors of Chagas disease. Infection, Genetics and Evolution 7 (2 (March)), 161167. Edman, J.D., 2003. Fitness advantages in multiple blood-feeding: the Aedes aegypti example. In: Takken, W., Scott, T.W. (Eds.), Ecological Aspects for Application of Genetically Modied Mosquitoes, vol. 2. Kluwer Academic. Wageningen UR frontis series, Dordrecht, ISSN 1573-4544, Chapter, 6374, 244 pp. Falconer, D.S., 1981. Introduction to Quantitative Genetics. Longman, London and New-York, 300 pp. Farris, J.S., 1972. Estimating phylogenetic trees from distance matrices. The American Naturalist 106, 645668. Fedor, P., Malenovsky, I., Vanhara, J., Sierka, W., Havel, J., 2008. Thrips (Thysanoptera) identication using articial neural network. Bulletin of Entomological Research 98, 437447. Feliciangeli, M., Sanchez-Martin, M., Marrero, R., Davies, C., Dujardin, J., 2007. Morphometric evidence for a possible role of Rhodnius prolixus from palm trees in house re-infestation in the State of Barinas (Venezuela). Acta Tropica 101, 169177. Fink, W.L., Zelditch, M.L., 1995. Phylogenetic analysis of ontogenetic shape transformations: a reassessment of the Piranha genus Pygocentrus (Teleostei). Systematic Biology 44, 344361. Fitzpatrick, S., Feliciangeli, M.D., Sanchez-Martin, M.J., Monteiro, F.A., Miles, M.A., 2008. Molecular genetics reveal that silvatic Rhodnius prolixus do colonise rural houses. PLoS Neglected Tropical Diseases 2 (4), e210. Foottit, R.G., Sorensen, J.T., 1992. Ordination methods: their contrast to clustering and cladistic techniques. In: Foottit, R.G., Sorensen, J.T. (Eds.), Ordination in the Study of Morphology, Evolution and Systematics of Insects: Applications and Quantitative Genetic Rationales. Elsevier, New York, pp. 110, 418 pp. Garcia, B., Moriyama, E., Powell, J., 2001. Mitochondrial DNA sequences of triatomines (Hemiptera: Reduviidae): Phylogenetic relationships. Journal of Medical Entomology 38 (5), 675683. Gilbert, S., 2000. Diachronics biology meets Evo-Devo: C.H. Waddingtons approach to Evolutionary Developmental Biology. American Zoologist 40 (5), 729737. Gorur, G., 2005. The importance of phenotypic plasticity in herbivorous insect speciation. In: Anantakrishnan, T.N., Whitman, D. (Eds.), Insect Phenotypic Plasticity. Diversity of Responses. Science Publishers, Inc., Eneld (NH), USA, pp. 145172. Graham, J., 1992. Genomic coadaptation and developmental stability in hybrid zones. In: Zakharov, V.M., Graham, J.H. (Eds.), Proceedings, Developmental Stability in Natural Populations, vol. 191. Acta Zoologica Fennica, pp. 121131. Graham, J., Freeman, D., Emlen, J., 1993. Developmental stability: a sensitive indicator of populations under stress. In: Wayne, G., Landis, Jane, S., Hugues, Michael, A., Lewis, (Eds.), Environmental Toxicology and Risk assessment, ASTM STP 1179. American Society for Testing and Materials, Philadelphia, pp. 136158. Gumiel, M., Catala, S., Noireau, F., de Arias, A.R., Garcia, A., Dujardin, J.P., 2003. Wing geometry in Triatoma infestans (Klug) and T. melanosoma Martinez, Olmedo and Carcavallo (Hemiptera: Reduviidae). Systematic Entomology 28 (2), 173179. Haramis, L., 1983. Increased adult size correlated with parity in Aedes triseriatus. Mosquito News 43, 7779. Harrington, L., Scott, T.W., Lerdthusnee, K., Coleman, R.C., Costero, A., Clark, G.G., Jones, J.J., Kitthawee, S., Kittayapong, P., Sithiprasasna, R., Edman, J.D., 2005. Dispersal of the dengue vector Aedes aegypti within and between rural communities. American Journal of Tropical Medicine and Hygiene 72 (2), 209220. Harry, M. Variabilite genetique de populations venezueliennes de Rhodnius spp, vectrices de Trypanosoma cruzi, parasite responsable de la maladie de Chagas. Ph.D Thesis, Universite Paris V, 1992. Hiernaux, J., 1977. Long-term biological effects of human migration from the african savanna to the equatorial forest: a case study of human adaptation to a hot and wet climate. In: Harrison, G.A. (Ed.), Population Structure and Human Variation. Cambridge University Press, Cambridge, pp. 187218. Honek, A., 1993. Intraspecic variation in body size and fecundity in insects: a general relationship. Oikos 66, 483492. Houle, D., 1992. Comparing evolvability and variability of quantitative traits. Genetics 130, 195204. Hugot, J.P., Baylac, M., 1996. Comparative landmark analysis of various oxyuridae parasites of primates and rodents. Patterns of variation seen in the Caudal Bursa of males. In: Marcus, L.F., Corti, M., Loy, A., Naylor, G.J.P., Slice, D. (Eds.), Advances in Morphometrics. Proceedings of the 1993 NATO-ASI on Morphometrics. Plenum Publ. NATO ASI, ser. A, Life Sciences, New York, pp. 463478. Hutcheson, H.J., Oliver, J.H., Houck, M.A., Strauss, R.E., 1995. Multivariate morphometric discrimination of nymphal and adult forms of the blacklegged tick (Acari: Ixodidae), a principal vector of the agent of Lyme disease in Eastern North America. Entomological Society of America 32 (6), 827842. Iriarte, F.P.W., Cespedes, W.C., Santos, M., 2003. Quantitative genetic analysis of wing form and bilateral asymmetry in isochromosomal lines of Drosophila subobscura using Procrustes methods. Journal of Genetics 82, 95113.
De Deken, R., van den Bossche, P., Sangare, M., Gnanvi, C., Missanda, J.H., Van Hees, J., 1997. Effect of the life-span of female Glossina palpalis gambiensis on the weight and size of its progeny. Medical and Veterinary Entomology 11 (1), 95101. De la Riva, J., Le Pont, F., Ali, V., Matias, R., Mollinedo, S., Dujardin, J., 2001. Wing geometry as a tool for studying the Lutzomyia longipalpis (Diptera: Psychodidae) complex. Memorias do Instituto Oswaldo Cruz 96 (8), 10891094. Debat, V., Milton, C.C., Rutherford, S., Klingenberg, C.P., Hoffmann, A.A., 2006. Hsp90 and the quantitative variation of wing shape in Drosophila melanogaster. Evolution 60, 25292538. Do, M.T., Harp, J.M., Norris, K.C., 1999. A test of a pattern recognition system for identication of spiders. Bulletin of Entomological Research 89, 217224. Dorn, P., Melgar, S., Rouzier, V., Gutierrez, A., Combe, C., Rosales, R., Rodas, A., Kott, S., Salvia, D., Monroy, C., 2003. The Chagas vector, Triatoma dimidiata (Hemiptera: Reduviidae), is panmictic within and among adjacent villages in Guatemala. Journal of Medical Entomology 40 (4), 436440. Duda, R.O., Hart, P.E., 1973. Pattern Classication and Scene Analysis. A WileyInterscience Publication, New York, p. 482. Dujardin, J.P., 1998. Population genetics and the natural history of domestication in Triatominae. Memorias do Instituto Oswaldo Cruz 93, 3436. Dujardin, J.P., Le Pont, F., 2000. Morphometrics of a neotropical sandy subspecies, Lutzomyia carrerai thula. Comptes-rendus de IAcademie des Sciences Serie III Sciences de la VieLife Sciences 323 (3), 273279. Dujardin, J.P., Le Pont, F., 2004. Geographic variation of metric properties within neotropical sandies. Infections, Genetics and Evolution 4 (4), 353359. Dujardin, J.P., Schoeld, C.J., 2004. Triatominae: systematics, morphology and population biology. In: Maudlin, I., Holmes, P.H., Miles, M. (Eds.), The Trypanosomiases. CAB International Oxfordshire, UK, pp. 181201, 614 pp. Dujardin, J.P., Schoeld, C.J., 2007. Vector control by surveillance networks: the ECLAT program and Chagas. In: Tibayrenc, M. (Ed.), Encyclopedia of Infectious Diseases. Modern Methodologies. Wiley & Sons, Chapter 24, pp. 425434. Dujardin, J.P., Slice, D., 2007. Geometric morphometrics. Contributions to medical entomology. In: Tibayrenc, M. (Ed.), Encyclopedia of Infectious Diseases. Modern Methodologies. Wiley & Sons, Chapter 25, pp. 435447. Dujardin, J.P., Cardozo, L., Schoeld, C.J., 1996a. Genetic analysis of Triatoma infestans following insecticidal control interventions in central Bolivia. Acta Tropica 61, 263266. Dujardin, J.P., Le Pont, F., Cruz, M., Leon, R., Guderian, T.F.R., Echeverria, R., Tibayrenc, M., 1996b. Cryptic speciation in Lutzomyia (Nyssomyia) trapidoi (Fairchild and Hertig) (Diptera: Psychodidae) detected by Multilocus Enzyme Electrophoresis. American Journal of Tropical Medicine and Hygiene 54 (1), 4245. Dujardin, J.P., Bermudez, H., Casini, C., Schoeld, C.J., Tibayrenc, M., 1997a. Metric differences between silvatic and domestic Triatoma infestans (Hemiptera, Reduviidae) in Bolivia. Journal of Medical Entomology 34 (5), 544552. Dujardin, J.P., Bermudez, H., Schoeld, C.J., 1997b. The use of morphometrics in entomological surveillance of silvatic foci of Triatoma infestans in Bolivia. Acta Tropica 66, 145153. Dujardin, J.P., Torrez, E.M., Le Pont, F., Hervas, D., Sossa, D., 1997c. Isozymic and metric variation in the Lutzomyia longipalpis complex. Medical and Veterinary Entomology 11, 394400. Dujardin, J.P., Munoz, M., Chavez, T., Ponce, C., Moreno, J., Schoeld, C.J., 1998a. The origin of Rhodnius prolixus in Central America. Medical and Veterinary Entomology 12, 113115. Dujardin, J.P., Schoeld, C.J., Tibayrenc, M., 1998b. Population structure of Andean Triatoma infestans: allozyme frequencies and their epidemiological relevance. Medical and Veterinary Entomology 12, 2029. Dujardin, J.P., Bermudez, H., Gianella, A., Cardozo, L., Ramos, E., Saravia, R., Quiroz, K., Forgues, G., Carazas, R., Hervas, D., Chavez, T., Machane, M., Martnez, E., Torrez, M., 1999a. Uso de marcadores geneticos en la vigilancia entomologica de la enfermedad de Chagas. In: Cassab, J.A., Noireau, F., Guillen, G. (Eds.), En: La Enfermedad de Chagas en Bolivia - Conocimientos cientcos al inicio del Programa de Control (19982002). Ministerio de Salud y Prevision social, OMS/OPS, IRD and IBBA, La Paz, pp. 157169, 259 pp. Dujardin, J.P., Chavez, T., Moreno, J.M., Machane, M., Noireau, F., Schoeld, C.J., 1999b. Comparison of isoenzyme electrophoresis and morphometric analysis for phylogenetic reconstruction of the Rhodniini (Hemiptera: Reduviidae: Triatominae). Journal of Medical Entomology 36, 653659. Dujardin, J.P., Le Pont, F., Galati, E.A.B., 1999c. Cryptic speciation suspected by morphometry in sandy (Diptera: Phlebotominae). Comptes Rendus de IAca demie des Sciences. Life Sciences 322, 375382. Dujardin, J.P., Le Pont, F., Martinez, E., 1999d. Quantitative morphological evidence for incipient species within Lutzomyia quinquefer (Diptera: Psychodidae). Memorias do Oswaldo Cruz 94 (6), 829836. Dujardin, J.P., Le Pont, F., Martinez, E., 1999e. Quantitative phenetics and taxonomy of some phlebotomine taxa. Memorias of Oswaldo Cruz 94 (6), 735741. Dujardin, J.P., Panzera, P., Schoeld, C.J., 1999f. Triatominae as a model of morpho logical plasticity under ecological pressure. Memorias do Instituto Oswaldo Cruz 94, 223228. Dujardin, J.P., Steindel, M., Chavez, T., Martinez, E., Schoeld, C.J., 1999g. Changes in the sexual dimorphism of Triatominae in the transition from natural to articial habitats. Memorias do Instituto Oswaldo Cruz 94, 565569. Dujardin, J.P., Schoeld, C.J., Panzera, F., 2002. Los Vectores de la Enfermedad de Chagas. Investigaciones taxonomicas, biologicas y geneticas. Academie Royale des Sciences dOutre-Mer, Classe des Sciences naturelles et medicales. Traduction espagnole.
J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890 Jablonka, E., Lamb, M., 2002. The changing concept of epigenetics. Annals of the New York Academy of Sciences 981, 8296. Jablonka, E., Lamb, M.J., Lachmann, M., 1992. Evidence, mechanisms and models for the inheritance of acquired characteristics. Journal of Theoretical Biology 158 (2), 245268. James, F.C., 1970. Geographic size variation in birds and its relationship to climate. Ecology 51, 365390. Jaramillo, N., Castillo, D., Wolff, M., 2002. Geometric morphometric differences between Panstrongylus geniculatus from eld and laboratory. Memorias do Instituto Oswaldo Cruz 97 (5), 667673. Jirakanjanakit, N. Geometric morphometrics of Aedes aegypti. PhD Thesis, 2007, 200 pp. Jirakanjanakit, N., Dujardin, J., 2005. Discrimination of Aedes aegypti (Diptera; Culicidae) laboratory lines based on wing geometry. The Southeast Asian Journal of Tropical Medicine and Public Health 36 (4), 14. Jirakanjanakit, N., Leemingsawat, S., Thongrungkiat, S., Apiwathnasorn, C., Singhaniyom, S., Bellec, C., Dujardin, J., 2007. Inuence of larval density or food variation on the geometry of the wing of Aedes (Stegomyia) aegypti. Tropical Medicine and International Health 12 (11), 13541360. Jirakanjanakit, N., Leemingsawat, S., Dujardin, J., 2008. The geometry of the wing of Aedes (Stegomyia) aegypti in isofemale lines through successive generations. Infect. Genet. Evol. 8 (4), 414421. Johansen, C.A., 1972. Toxicity of eld-weathered insecticide residues to four kinds of bees. Environmental Entomology 1, 393394. Jones, T.M., Quinnell, R.J., 2002. Testing predictions for the evolution of lekking in the sandy, Lutzomyia longipalpis. Animal Behaviour 63 (3), 605612. Jordaens, K., Van Dongen, S., Van Riel, P., Geenen, S., Verhagen, R., Backeljau, T., 2002. Multivariate morphometrics of soft body parts in terrestrial slugs: comparison between two datasets, error assessment and taxonomic implications. Biological Journal of the Linnean Society. APR 75 (4), 533542. Kendall, D.G., 1986. Comments on f.l. bookstein, size and shape spaces for landmark data in two dimensions. Statistical Sciences 1, 222226. Kitthawee, S., Edman, J.D., Upatham, E.S., 1992. Relationship between female Anopheles dirus (Diptera: Culicidae) body size and parity in a biting population. Journal of Medical Entomology 29, 921926. Klingenberg, C., Leamy, L.J., Cheverud, J.M., 2004. Integration and modularity of quantitative trait locus effects on geometric shape in the mouse mandible. Genetics 166, 19091921. Klingenberg, C.P., 1996. Multivariate allometry. In: Marcus, L.F., Corti, M., Loy, A., Naylor, G.J.P., Slice, D. (Eds.), Advances in Morphometrics Proceedings of the 1993 NATO-ASI on Morphometrics. Plenum Publ. NATO ASI, seR. A, Life Sciences, New York, pp. 2349. Klingenberg, C.P., Leamy, L.J., 2001. Quantitative genetics of geometric shape in the mouse mandible. Evolution 55, 23422352. Klingenberg, C.P., Spence, J.R., 1997. On the role of body size for life-history evolution. Ecological Entomology 22, 5568. Klingenberg, C.P., Leamy, L.J., Routman, E.J., Cheverud, J.M., 2001. Genetic architecture of mandible shape in mice: effects of quantitative trait loci analyzed by geometric morphometrics. Genetics 157 (2), 785802. Klingenberg, C.P., Barluenga, M., Meyer, A., 2002. Shape analysis of symmetric structures: quantifying variation among individuals and asymmetry. Evolution 56 (10), 19091920. Klowden, M.J., Blackmer, J.L., Chambers, G.M., 1988. Effects of larval nutrition on the host-seeking behaviour of adult Aedes aegypti mosquitoes. Journal of the American Mosquito Control Association 4, 7375. Lane, R.P., Marshall, J., 1981. Geographical variation, races and subspecies. In: Forey, P.L. (Ed.), The Evolving Biosphere. British Museum (Natural History), London, pp. 919. Lane, R.P, Ready, P.D., 1985. Multivariate discrimination between Lutzomyia wellcomei, a vector of mucocutaneous leishmaniasis, and Lu. complexus (Diptera: Phlebotominae). Annals of Tropical Medicine and Parasitology 79 (4), 469 472. Largiader, C., Klingenberg, C., Zimmermann, M., 1994. Morphometric variation in a hybrid zone of two subspecies of Gerris costae (Heteroptera: Gerridae) in the Maritime Alps. Journal of Evolution Biology 7, 697712. Le Pont, F., Martinez, E.M., Torrez, M., Dujardin, J.P., 1997a. Description de 5 ` ` nouvelles especes de Lutzomyia (Diptera: Psychodidae) de region subandine. Bulletin de la Societe Entomologique de France 103 (2), 159173. Le Pont, F., Torrez-Espejo, M., Dujardin, J.P., 1997b. Phlebotomes de Bolivie: description de quatre nouvelles especes de Lutzomyia (Diptera: Psychodidae). Ann Soc Entomol Fr 33, 5564. Le Sueur, D., Sharp, B.L., 1991. Temperature-dependent variation in Anopheles merus larval head capsule width and adult wing length: implication for anopheline taxonomy. Medical and Veterinary Entomology 5, 5562. Leahy, M.G., Vandehey, R.C., Booth, K.S., 1978. Differential response to oviposition site by feral and domestic populations of Aedes aegypti (L.) (Diptera: Culicidae). Bulletin of Entomological Research 68, 455463. Lehmann, T., Dalton, R., Kim, E., Dahl, E., Diabate, A., Dabire, R., Dujardin, J., 2006. Genetic contribution to variation in larval development time, adult size, and longevity of starved adults of Anopheles gambiae. Infection, Genetics and Evolution 6 (5), 410416. Lele, S.R., Richtsmeyer, J., 2001. An Invariant Approach to Statistical Analysis of Shape. Chapman and Hall/CRC, Boca Raton, pp. VIII+308. Levit, G.S., Hossfeld, U., Olssonza, L., 2006. From the Modern Synthesis to Cybernetics: Ivan Ivanovich Schmalhausen (18841963) and his research pro-
889
gram for a Synthesis of Evolutionary and Developmental Biology. Journal of Experimental Zoology (Mol Dev Evol) 306B, 89106. Long, A., Mullaney, S., Reid, L., Fry, J., Langley, C., Mackay, T.F.C., 1995. High resolution mapping of genetic factors affecting abdominal bristle number in Drosophila melanogaster. Genetics 139 . Lyimo, E., Takken, W., 1993. Effects of adult body size on fecundity and the pregravid rate of anopheles gambiae females in Tanzania. Medical and Veterinary Entomology 7, 328332. Maciel de Freitas, R., Codeco, C.T., Lourenco de Oliveira, R., 2007. Body sizeassociated survival and dispersal rates of Aedes aegypti in Rio de Janeiro. Medical and Veterinary Entomology 21, 284292. MacLeod, N., 2002. Phylogenetic signals in morphometric data. In: MacLeod, N., Forey, P.L. (Eds.), Morphology, Shape and Phylogeny. Syst. Ass. Spec. Vol. Ser. 64. Taylor and Francis, London, 100138, 308 pp. MacLeod, N., ONeill, M., Walsh, S.A., 2007. A comparison between morphometric and articial neural network approaches to the automated species recognition problems in systematics. In: Curry, G.B., Humphries, C.J. (Eds.), Biodiversity Databases: Techniques, Politics, and Applications (Systematics Association Special Volume) - US. 9780415332903Chapter V, pp. 3762. Malmqvist, B., Adler, P.H., Strasevicius, D., 2004. Testing hypotheses on egg number and size in black ies (Diptera: Simuliidae). Journal of Vector Ecology 2 (29), 248256. Manoukis, N.C., Toure, M.B., Sissoko, I., Doumbia, S., Traore, S.F., Diuk-Wasser, M.A., Taylor, C.E., 2006. Is vector body size the key to reduced malaria transmission in the irrigated region of Niono, Mali? Journal of Medical Entomology 43 (5), 820 827. Marcondes, C.B., Borges, P.S.S., 2000. Distinction of males of the Lutzomyia intermedia (Lutz & Neiva, 1912) species complex by ratios between dimensions and by an articial neural network (Diptera: Psychodidae, Phlebotominae). Memorias of Oswaldo Cruz 95 (5), 685688. Marcondes, C.B., Lozovei, A.L., Galati, E.A.B., Taniguchi, H.H., 1998. The usefulness of Bergmanns rule for the distinction of members of Lutzomyia intermedia species complex (Diptera, Psychodidae, Phlebotominae). Memorias do Instituo Oswaldo Cruz, Rio de Janeiro 93, 363364. Martinez, E., Le Pont, F., Torrez, M., Tellera, J., Vargas, F., Dujardin, J., Dujardin, J.P., 1999. Lutzomyia nuneztovari anglesi (Le Pont and Desjeux, 1984) as a vector of Leishmania ama-zonensis in a sub-andean leishmaniasis focus of Bolivia. American Journal of Tropical Medicine and Hygiene 5, 846849. Matias, A., De la Riva, J.X., Martinez, E., Dujardin, J.P., 2002. Domiciliation process of Rhodnius stali (Hemiptera: Reduviidae) in the Alto Beni (La Paz, Bolivia). Tropical Medicine and International Health 8 (3), 264268. McKay, J.K., Latta, R.G., 2002. Adaptive population divergence: markers, QTL and traits. Trends in Ecology and Evolution 17, 285291. McKenzie, J., Clarke, G., 1988. Diazinon resistance, uctuating asymmetry and tness in the Australian sheep blowy, Lucilia cuprina. Genetics 120 (1), 213220. Milton, C.C., Huynh, B., Batterham, P., Rutherford, S.L., Hoffmann, A.A., 2003. Quantitative trait symmetry independent of hsp90 buffering: distinct modes of genetic canalization and developmental stability. Proceedings of the National Academy of Sciences of the United States of America 100 (23), 1339613401. Moller, A., Swaddle, J., 1997. Asymmetry, developmental stability, and evolution. Oxford Series in Ecology and Evolution 1291. Mller, A.P., 1992. Parasites differentially increase the degree of uctuating asymmetry in secondary sexual characters. Journal of Evolution Biology 5, 691699. Mousseau, A.P., Roff, D.A., 1987. Natural selection and the heritability of tness. Heredity 59, 181197. Mpho, M., Holloway, G., Callaghan, A., 2000. Fluctuating wing asymmetry and larval density stress in Culex quinquefasciatus (Diptera: Culicidae). Bulletin of Entomological Research 90 (3), 279283. Mwangangi, J.M., Mbogo, C.M., J G.u, Kabiru, E.W., Mwambi, H., Githure, J.I., Beyer J. C., 2004. Relationships between body size of anopheles mosquitoes and Plasmodium falciparum sporozoite rates along the Kenya coast. Journal of America Mosquito Control Association 20, 390394. Nijhout, H.F., 2003. The control of body size in insects. Developmental Biology 261, 19. Noireau, F., Menezes dos Santos, S., Gumiel, M., Dujardin, J., dos Santos Soares, M., Car-cavallo, R.U., Galvao, C., Jurberg, J., 2002. Phylogenetic relationships within the oliveirai complex (hemiptera: Reduviidae: Triatominae). Infection, Genetics and Evolution 2, 1117. Palmer, A.R., Strobeck, C., 1986. Fluctuating asymmetry: measurement, analysis, patterns. Annual review of Ecology and Systematics 17, 391421. Parsons, P.A., 1990. Fluctuating asymmetry: an epigenetic measure of stress. Biological reviews of the Cambridge Philosophical Society 65, 131145. Partridge, L., Barrie, B., Fowler, K., French, V., 1994. Evolution and development of body size and cell size in Drosophila melanogaster in response to temperature. Evolution 48, 12691276. Patterson, J., Klingenberg, C., 2007. Developmental buffering: how many genes? Evolution and Development 9 (6), 525526. Pennisi, E., 2006. Evolution: hidden genetic variation yields caterpillar of a different color. Science 591a . Periago, M., Valero, M.A., El Sayed, M., Ashra, K., El Wakeel, A., Mohamed, M.Y., Desquesnes, M., Curtale, F., Mas-Coma, S., 2008. First phenotypic description of Fasciola hep-atica/Fasciola gigantica intermediate forms from the human endemic area of the nile delta, egypt. Infections, Genetics and Evolution 8 (1), 5158.
890
J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890 Spitze, K., 1993. Population structure in Daphnia obtusa: quantitative genetic and allozymic variation. Genetics 135, 367374. Strickman, D., Kittayapong, P., 2003. Dengue and its vectors in Thailand: calculated transmission risk from total pupal counts of Aedes aegypti and association of wing-length measurements with aspects of the larval habitat. American Journal of Tropical Medicine and Hygiene 68 (2), 209217. Sukonthabirhom, S., Rongnoparut, P., Saengtharatip, S., Jirakanjanakit, N., Chareonviriyaphap, T., 2005. Genetic structure and gene ow among Aedes (Stegomyia) aegypti (Diptera: Culi-cidae) populations from Central Thailand. Journal of Medical Entomology 42 (4), 604609. Suzuki, Y., Nijhout, H.F., 2006. Evolution of a polyphenism by genetic accommodation. Science 5761 (311), 650652. Swiderski, D.L., Zelditch, M.L., Fink, W.L., 2002. Comparability, morphometrics and phylogenetic systematics. In: MacLeod, N., Forey, P.L. (Eds.), Morphology, Shape and Phylogeny. Syst. Ass. Spec. Vol. Ser. 64. Taylor and Francis, London, pp. 6799. Takken, W., Klowden, M.J., Chambers, G.M., 1998. Effect of body size on host seeking and blood meal utilization in Anopheles gambiae sensu stricto (Diptera: Culicidae): the disadvantage of being small. Journal of Medical Entomology 35, 639645. Tapis, M., Hausermann, W., 1978. Genetics of house entering behaviour in east african populations of Aedes aegypti (L.) (Diptera: Culicidae) and its relevance to speciation. Bulletin of Entomological Research 68, 521532. Tapis, M., Hausermann, W., 1975. Demonstration of differential domesticity of Aedes aegypti (L.) (Diptera: Culicidae) in Africa by mark-release-recapture. Bulletin of Entomological Research 65, 199208. Thackeray, J.F., Bellamy, C.L., Bellars, D., Bronner, G., Bronner, L., Chimimba, C., Fourie, H., Kemp, A., Krueger, M., Plug, I., Prinsloo, S., Toms, R., Van Zyl, A.J., Whiting, M.J., 1997. Probabilities of conspecicity: application of a morphometric technique to modern taxa and fossil specimens attributed to Australopithecus and Homo. South African Journal of Science 93 (4), 195196. Tolski, A., 2004. Drawwing, a program for numerical description of insect wings. Journal of Insect Science 4, 17. Torrez, M., Lopez, M., Le Pont, F., Martinez, E., Munoz, M., Hervas, D., Yaksic, N., Arevalo, J., Sossa, D., Dujardin, J., 1999. Lutzomyia nuneztovari anglesi (Diptera: Psychodidae) as a probable vector of L. b. braziliensis in the Yungas (Bolivia). Acta Tropica 71, 311316. Van Valen, L., 1962. A study of uctuating asymmetry. Evolution 16, 125142. Vanhara, J., Murarikova, N., Malenovsky, I., Havel, J., 2007. Articial neural networks for y identication: a case study from the genera Tachina and Ectophasia (Diptera, Tachinidae). Biologia 62, 462469. Vanvoorhies, W., 1996. Bergmann size clinesa simple explanation for their occurrence in ectotherms. Evolution 50 (3), 12591264. Villegas, J., Feliciangeli, M.D., Dujardin, J.P., 2002. Wing shape divergence between Rhodnius pro-lixus from Cojedes (Venezuela) and R. robustus from Merida (Venezuela). Infection, Genetics and Evolution 2, 121128. Waddington, C.H., 1953. Genetic assimilation for an acquired character. Evolution 7, 118126. Waddington, C.H., 1956. Genetic assimilation of the bithorax phenotype. Evolution 10, 113. Ward, R.D., Phillips, A., Burnet, B., Marcondes, C.B., 1988. The Lutzomyia longipalpis complex: reproduction and distribution. In: Service, M.W. (Ed.), Biosystematics of Haematophagous Insects, 37. Systematics Association Special, 257269, 376 pp. West-Eberhard, M., 2003. Developmental Plasticity and Evolution. Oxford University Press, 794 pp. Whitlock, M.C., 1999. Neutral additive variance in a metapopulation. Genetic Research 74, 215221. Wiwatanaratanabutra, S., Kittayapong, P., 2006. Effects of temephos and temperature on Wolbachia load and life history traits of Aedes albopictus. Medical and Veterinary Entomology 20 (3), 300307. Workman, M.S., Leamy, L.J., Routman, E.J., Cheverud, J.M., 2002. Analysis of quantitative trait locus effects on the size and shape of mandibular molars in mice. Genetics 160 (4), 15731586. Xue, R.d., Edman, J., Scott, T., 1995. Age and body size effects on blood meal size and multiple blood feeding by Aedes aegypti (Diptera: Culicidae). Journal of Medical Entomology 32 (4), 471474. Young, D.G., Duncan, M.A., 1994. Guide to the identication and geographic distribution of Lutzomyia sand ies in Mexico, the West Indies, Central and South America (Diptera: Psy-chodidae). Associate Publishers, Gainesville, Fl, pp. 881. Zelditch, M.L., Fink, W.L., Swiderski, D.L., 1995. Morphometrics, homology, and phylogenetics: quantied characters as synapomorphies. Systematic Biology 44 (2), 179189. Zeledon, R. El Triatoma dimidiata y su relacion con la enfermedad de Chagas. Editorial Universidad Estatal a Distancia (EUNED), San Jose, Costa Rica, 1981. Zeledon, R., Guardia, V., Zuinga, A., Swartzwelder, J., 1970. Biology and ethology of Triatoma dimidiata (Latreille, 1811). I. Life cycle, amount of blood ingested, resistance of starvation, and size of adults. Journal of Medical Entomology 7, 313319.
Pigliucci, M., 2005. Evolution of phenotypic plasticity: where are we going now? Trends in Ecology and Evolution 20 (9), 481486. Pimentel, R.A., 1992. An introduction to ordination, principal components analysis and discriminant analysis. In: Foottit, R.G., Sorensen, J.T. (Eds.), Ordination in the Study of Morphology, Evolution and Systematics of Insects: Applications and Quantitative Genetic Rationales. Elsevier, New York, pp. 1128, pp. 418. Platnick, N., Russel, K.N., Do, M.T. Spida-Web Species Identied Automatically. A neural network based automated identication system for biological species. https://research.amnh.org/invertzoo/spida/common/index.htm, 2005. Pomiankowski, A., 1997. Genetic variation in uctuating asymmetry. Journal of Evolution Biology 10, 5155. Rasmussen, P., Wheeler, W., Moser, T., Vine, L., Sullivan, B., Rusch, D., 2001. Measurements of Canada goose morphologysources of error and effects on classication of subspecies. Journal of Wildlife Management 65 (4), 716725. Ray, C., 1960. The application of Bergmanns and Allens rules to the poikilotherms. Journal of Morphology 106, 85108. Rodrguez, J., Gonzalez, O., Nodarze, J., Fidalgo, L., Dujardin, J., 2007. Morphometric changes of Triatoma avida Neiva, 1911 (Hemiptera: Triatominae) in the transition from sylvatic to laboratory condition. Revista do Instituto de Med icina Tropical de Sao Paulo 49, 127130. Rogers, A., Harpending, H.C., 1983. Population structure and quantitative characters. Genetics 105, 9851002. Rohlf, F.J., 1998. On applications of geometric morphometrics to studies of ontogeny and phylogeny. Systematic Biology 47, 147158. Rohlf, F.J., 1996. Morphometric spaces, shape components and the effects of linear transformations. In: Marcus, L.F., Corti, M., Loy, A., Naylor, G., Slice, D. (Eds.), Advances in Morphometrics. Proceedings of the 1993 NATO-ASI on Morphometrics. Plenum Publ. NATO ASI, ser. A, Life Sciences, New York, pp. 117129. Rohlf, F.J., 1990. Rotational t (Procrustes) methods. In: Rohlf, F., Bookstein, F. (Eds.), Proceedings of the Michigan Morphometrics Workshop. Special Publication Number 2. The University of Michigan Museum of Zoology, Ann Arbor, MI, pp. 227236, 380 pp. Rohlf, F.J., Bookstein, F.L., 2003. Computing the uniform component of shape variation. Systematic Biology 52 (1), 6669. Rohlf, F.J., Marcus, L.F., 1993. A revolution in morphometrics. TREE 8 (4), 129132. Rohlf, F.J., Loy, A., Corti, M., 1996. Morphometric analysis of Old World Talpidae (Mammalia, Insectivora) using partial-warp scores. Systematic Biology 45 (3), 344362. Rutherford, S.L., Lindquist, S., 1998. Hsp90 as a capacitor for morphological evolution. Nature 396, 336342. Schilthuizen, M., 2000. Dualism and conicts in understanding speciation. BioEssays 22, 11341141. Schneider, J.R., Mori, A., Romero-Severson, J., Chadee, D.D., Severson, D.W., 2007. Investigations of dengue-2 susceptibility and body size among Aedes aegypti populations. Medical and Veterinary Entomology 4 (21), 370376. Schneider, S.S., Leamy, L.J., Lewis, L.A., DeGrandi-Hoffman, G., 2003. The inuence of hybridization between African and European honeybees, Apis mellifera, on asymmetries in wing size and shape. Evolution 57 (10), 23502364. Schoeld, C., Patterson, J., 2005. Preparing for tsetse eradication. South African Journal of Science 101, 116. Schoeld, C.J., Dias, J.C.P., 1998. The southern cone initiative against Chagas disease. Advances in Parasitology 42, 127. Schoeld, C.J., Dias, J.P.C., 1991. A cost-benet analysis of Chagas disease control. Memorias do Instituo Oswaldo Cruz, Rio de Janeiro 86 (3), 285295. Schoeld, C.J., Diotaiuti, L., Dujardin, J.P., 1999. The process of domestication in Triatominae. Memorias of Instituto Oswaldo Cruz 94 (Suppl. I), 375378. Scoble, M.J., Clark, B.R., Godfray, H.C.J., Kitching, I.J., Mayo, S.J., 2007. Revisionary taxonomy in a changing e-landscape. Tijdschrift voor Entomologie 150, 305317. Shrimpton, A., Robertson, A., 1988. The isolation of polygenic factors controlling bristle score in Drosophila melanogaster. I. Allocation of third chromosome bristle effects to chromosome sections. Genetics 118, 437443. Siegel, J., Novak, R., Lampman, R.L., Steinly, B.A., 1992. Statistical appraisal of the weight-wing length relationship of mosquitoes. Journal of Medical Entomology 29 (4), 711714. Slatkin, M., 1985. Rare alleles as indicators of gene ow. Evolution 39 (1), 5365. Slatkin, M., 1981. Estimating levels of gene ow in natural populations. Genetics 99, 323335. Sollars, V., Lu, X., Xiao, L., Wang, X., Garnkel, M., Ruden, D.M., 2002. Evidence for an epigenetic mechanism by which Hsp90 acts as a capacitor for morphological evolution. Nature Genetics 33, 7074. Sorensen, J., 1992. The use of discriminant function analysis for estimation of phylogeny: partitioning, perspective and problems. In: Foottit, R.G., Sorensen, J.T. (Eds.), Ordination in the Study of Morphology, Evolution and Systematics of Insects: Applications and Quantitative Genetic Rationales. Elsevier, New York, pp. 6593, 418 pp. Soule, M.E., 1979. Heterozygosity and developmental stability: another look. Evolution 33 (1), 396401.

2008 - Dujardin - Morphometrics Applied To Mediacal Entomology

Hochgeladen von

Dokumentinformationen

Originalbeschreibung:

Originaltitel

Copyright

Verfügbare Formate

Dieses Dokument teilen

Dokument teilen oder einbetten

Freigabeoptionen

Stufen Sie dieses Dokument als nützlich ein?

Sind diese Inhalte unangemessen?

Copyright:

Verfügbare Formate

2008 - Dujardin - Morphometrics Applied To Mediacal Entomology

Hochgeladen von

Copyright:

Verfügbare Formate

Infection, Genetics and Evolution 8 (2008) 875890

Contents lists available at ScienceDirect

Infection, Genetics and Evolution

Morphometrics applied to medical entomology

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

0.77 (0.06) 0.68 (0.06) 0.45 (0.02)

0.71 (0.01) 0.35 (0.01)

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

J.-P. Dujardin / Infection, Genetics and Evolution 8 (2008) 875890

Das könnte Ihnen auch gefallen