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      Wing morphometrics as a tool in species identification of forensically important blow flies of Thailand

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          Abstract

          Background

          Correct species identification of blow flies is a crucial step for understanding their biology, which can be used not only for designing fly control programs, but also to determine the minimum time since death. Identification techniques are usually based on morphological and molecular characters. However, the use of classical morphology requires experienced entomologists for correct identification; while molecular techniques rely on a sound laboratory expertise and remain ambiguous for certain taxa. Landmark-based geometric morphometric analysis of insect wings has been extensively applied in species identification. However, few wing morphometric analyses of blow fly species have been published.

          Methods

          We applied a landmark-based geometric morphometric analysis of wings for species identification of 12 medically and forensically important blow fly species of Thailand. Nineteen landmarks of each right wing of 372 specimens were digitised. Variation in wing size and wing shape was analysed and evaluated for allometric effects. The latter confirmed the influence of size on the shape differences between species and sexes. Wing shape variation among genera and species were analysed using canonical variates analysis followed by a cross-validation test.

          Results

          Wing size was not suitable for species discrimination, whereas wing shape can be a useful tool to separate taxa on both, genus and species level depending on the analysed taxa. It appeared to be highly reliable, especially for classifying Chrysomya species, but less robust for a species discrimination in the genera Lucilia and Hemipyrellia. Allometry did not affect species separation but had an impact on sexual shape dimorphism.

          Conclusions

          A landmark-based geometric morphometric analysis of wings is a useful additional method for species discrimination. It is a simple, reliable and inexpensive method, but it can be time-consuming locating the landmarks for a large scale study and requires non-damaged wings for analysis.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s13071-017-2163-z) contains supplementary material, which is available to authorized users.

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          Most cited references39

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          Forensic entomology: applications and limitations.

          Forensic entomology is the science of collecting and analysing insect evidence to aid in forensic investigations. Its main application is in the determination of the minimum time since death in cases of suspicious death, either by estimating the age of the oldest necrophagous insects that developed on the corpse, or by analysing the insect species composition on the corpse. In addition, toxicological and molecular examinations of these insects may help reveal the cause of death or even the identity of a victim, by associating a larva with its last meal, for example, in cases where insect evidence is left at a scene after human remains have been deliberately removed. Some fly species can develop not only on corpses but on living bodies too, causing myiasis. Analysis of larvae in such cases can demonstrate the period of neglect of humans or animals. Without the appropriate professional collection of insect evidence, an accurate and convincing presentation of such evidence in court will be hampered or even impossible. The present paper describes the principles and methods of forensic entomology and the optimal techniques for collecting insect evidence.
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            Morphometrics applied to medical entomology.

            Morphometrics underwent a revolution more than one decade ago. In the modern morphometrics, the estimate of size is now contained in a single variable reflecting variation in many directions, as many as there are landmarks under study, and shape is defined 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, significant biological and epidemiological features can be quantified more accurately. We discuss the evolutionary significance 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 deficiencies 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 flow levels among interbreeding populations are commonly used, but gene flow does not necessarily mean the current flow of migrants. Methods using the morphometric variation are neither suited to evaluate gene flow, nor are they adapted to estimate the flow 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 flow 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.
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              Morphometric tools for landmark data: geometry and biology

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                Author and article information

                Contributors
                narinsontigun@gmail.com
                kabkaew.s@cmu.ac.th
                zajac@med.uni-frankfurt.de
                zehner@em.uni-frankfurt.de
                kom.s@cmu.ac.th
                kdantra@gmail.com
                amendt@em.uni-frankfurt.de
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                10 May 2017
                10 May 2017
                2017
                : 10
                : 229
                Affiliations
                [1 ]ISNI 0000 0000 9039 7662, GRID grid.7132.7, Department of Parasitology, Faculty of Medicine, , Chiang Mai University, ; Chiang Mai, 50200 Thailand
                [2 ]Institute of Legal Medicine, Forensic Biology/Entomology, Kennedyallee 104, 60596 Frankfurt am Main, Germany
                Article
                2163
                10.1186/s13071-017-2163-z
                5424331
                28486970
                756a606f-dc01-49f6-ba9b-3eef4a2cea4f
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 9 December 2016
                : 1 May 2017
                Funding
                Funded by: Royal Golden Jubilee Ph.D. Program
                Award ID: PHD/0118/2556
                Award Recipient :
                Funded by: Faculty of Medicine Endowment Fund for Medical Research, Faculty of Medicine, Chiang Mai University
                Categories
                Research
                Custom metadata
                © The Author(s) 2017

                Parasitology
                species identification,forensic entomology,wing morphometry,blow fly,thailand
                Parasitology
                species identification, forensic entomology, wing morphometry, blow fly, thailand

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