research article the first survey of forensically...

Research ArticleThe First Survey of Forensically Important EntomofaunaCollected from Medicolegal Autopsies in South Korea

Sang Eon Shin1 Hyun Ju Lee1 Ji Hye Park1 Kwang Soo Ko1 Yu-Hoon Kim2

Kyung Ryoul Kim2 and Seong Hwan Park1

1Department of Legal Medicine Korea University College of Medicine 73 Inchon-ro Seongbuk-gu Seoul 136-705 Republic of Korea2National Forensic Service 10 Ipchun-ro Wonju-si Gangwon-do 220-170 Republic of Korea

Correspondence should be addressed to Seong Hwan Park kuforengmailcom

Received 3 April 2015 Revised 12 June 2015 Accepted 14 June 2015

Academic Editor Christen Rune Stensvold

Copyright copy 2015 Sang Eon Shin et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Forensic entomology applies insect evidence to legal problems such as the estimation of minimum postmortem interval (mPMI)For this purpose knowledge of the insect fauna that are attracted to human cadavers in each geographic region is a prerequisiteDespite many studies investigating the insect fauna attracted to meat there has been no survey of the entomofauna on humancadavers in the East Asian temperate climate zone particularly in Korea Therefore this study reports the entomofauna collectedfrom medicolegal autopsies in northeastern Seoul and its suburbs Insect samples were collected from 35 medicolegal autopsiesin 2010 2011 and 2013 Molecular and morphological methods were utilized for taxonomic identification Among 1398 individualsamples belonging to 3 orders 13 families 18 genera and 32 species the dominant family and species were Calliphoridae and Luciliasericata respectively Despite its limited scale this study provides a snapshot of the general entomofauna that are attracted to humancadavers in this region

1 Introduction

Medicolegal entomology which applies insect evidence tocrime scene investigations is one of the main fields offorensic entomology [1] Traditional methods that use earlypostmortem phenomena such as postmortem body coolinglivor mortis and rigor mortis to estimate the postmorteminterval (PMI) become invalid once postmortem phenomenasuch as autolysis and putrefaction occur [2] Evidence derivedfrom insects that visit a dead body is an alternative strategythat can be used to estimate the minimum PMI (mPMI)once the early postmortem phenomena become invalid Toestimate mPMI using insect evidence knowledge of theentomofauna that are attracted to dead bodies in a specificgeographic region is critical as is the knowledge of the growthrates of larvae of dominant carrion insects [1]Therefore sev-eral researchers have investigated the entomofauna that areattracted to human cadavers in various geographic regions[3ndash5] In Korea and neighboring East Asian countries withina similar temperate climate zone theDNAof dipteran speciesthat are attracted to pork liver baits or pig carcasses has been

analyzed for species identification [6ndash11] However there hasbeen no survey of carrion insects visiting human cadavers inthe temperate climate zone of East Asia despite medicolegalautopsies being quite actively performed In this study 35medicolegal autopsies of insect infestation were investigatedin the northeastern area of Seoul and its suburbs over threeyears to determine the forensically important entomofauna ofthis region

2 Materials and Methods

21 Sampling Insect infestation was observed in 35 medi-colegal autopsies in northeastern Seoul and its suburbsduring 2010 2011 and 2013 With the consent of the foren-sic pathologist who performed the autopsy about 40ndash50individual insects were collected from each cadaver Whenthere are numerous maggots and small numbers of otherinsects maggots and other insects were collected randomlyand totally respectively If there were very small numbersof individual insects on a cadaver all of the individualinsects were collected Approximately half of the livemaggots

Hindawi Publishing CorporationBioMed Research InternationalVolume 2015 Article ID 606728 6 pageshttpdxdoiorg1011552015606728

2 BioMed Research International

Table 1 Primer sequences used for the amplification and sequencing of dipteran COI genes

Name Sequence Binding siteF1 51015840-CCTTTAGAATTGCAGTCTAATGTCA-31015840 tRNA-cysteineF2 51015840-GGAGGATTTGGAAATTGATTAGTTCC-31015840 220ndash245 on COIF3 51015840-CTGCTACTTTATGAGCTTTAGG-31015840 1000ndash1022 on COIR1 51015840-CCTAAATTTGCTCATGTTGACA-31015840 2ndash23 on 119862119874119868119868lowast

R2 51015840-CAAGTTGTGTAAGCATC-31015840 1327ndash1343 on COIR3 51015840-CCAAAGAATCAAAATAAATGTTG-31015840 688ndash710 on COIlowast119862119874119868119868 cytochrome c oxidase subunit II gene

collected were preserved in an 80 ethanol solution at 4∘Cand the other half of the live maggots collected were sent tothe incubation chambers for rearing andmorphological taxo-nomic identification Nondipteran samples were immediatelypreserved in 80 ethanol solution without rearing All thesamples immersed in 80 ethanol solution were immediatelysent to a freezer (minus20∘C) Although the best method of killingand preservation of live maggots is treatment in boiling waterand then transfer to 80 ethanol solution to avoid browningand shrinkage of maggots [12 13] the process of immersionin boiling water was omitted because the purpose of thepreservation was merely DNA barcoding

22 Taxonomic Identification Morphological features ofadult individualswere observed under an SZX10 stereomicro-scope (Olympus Corporation Tokyo Japan) Maggots usedfor rearing were taxonomically identified to the species levelafter emergence [14ndash17] After initial identification at a familylevel on the basis of the shape of the posterior spiracles amaggot sample was used for DNA barcoding All the adultbeetles and beetle larvae were taxonomically identified on thebasis of their morphological features to the species and thefamily level respectively [18ndash21]

23 DNA Barcoding of Maggots DNA barcoding of maggotswas performed utilizing the mitochondrial cytochrome coxidase subunit I gene (COI) Polymerase chain reaction wasperformed mainly to amplify the barcoding regions locatedwithin the upstream third portion of COI Primer sequenceswhichwere adopted fromaprevious study are listed inTable 1[6] After automatic sequencing using an ABI Prism 3720xlgenetic analyzer (Applied Biosystems Foster City CA USA)nucleotide sequences were aligned and compared with pre-existing nucleotide sequences using MEGA6 Software [22]

24 Rearing Conditions for Maggots Because the identifica-tion of maggot species usingmorphologic features is difficultspecimens were reared to the adult stage inside incubationchambers under constant conditions (25∘C relative humidity70) The light and dark ratio of each chamber was set at16 8 Live maggots were placed on 5-6 g of fresh pork liver ina plastic cylinder measuring 95mm in height and diameterwith an adequate amount of sawdust under the pork liver

3 Results and Discussion

Weather records for the study period confirmed that north-eastern Seoul belongs to the temperate climate zone having

00

1000

2000

3000

4000

5000

6000

00

100

200

300

400

Jan

Feb

Mar

Apr

May

Jun Jul

Aug

Sep

Oct

Nov

Dec

Rain

fall

(mm

)

Rainfall (mm)Mean

MaximumMinimum

minus200

minus100

Air

tem

pera

ture

(∘C)

Figure 1 A climograph of mean maximum and minimum airtemperatures (∘C) and rainfall (mm) at Seoul weather station(37∘341015840N 126∘581015840E) in 2010 2011 and 2013 demonstrates hot humidsummers and cold dry winters

0

24

6

8

10

12

14

Jan

Feb

Mar

Apr

May

Jun Jul

Aug

Sep

Oct

Nov

Dec

0002 1

4

13

9

221 1

Num

ber o

f aut

opsy

case

s

Month

Figure 2 Medicolegal autopsy cases showing insect infestation inthe study region in 2010 2011 and 2013 were most frequent in thesummer (June July and August)

humid hot summers and dry cold winters with interveningmoderate springs and falls (Figure 1) Insect infestationof human cadavers was most common (26 autopsies) insummer (June July and August) and was not observedfrom January to March (Figure 2) Among 1398 individualsamples a total of 3 orders 13 families 18 genera and

BioMed Research International 3

Calliphoridae (987) 706

Sarcophagidae (102) 73

Piophilidae (89) 64

Stratiomyidae (6) 04Muscidae (3) 02 Phoridae (2) 01Dermestidae (91) 65

Silphidae (89) 64

Cleridae (18) 13Histeridae (5) 04 Staphylinidae (3) 02

Pteromalidae (2) 01

Nitidulidae (1) 01

Figure 3 Individual numbers of collected insects in each family among 1398 insect samples

32 species were identified The three orders identified wereDiptera (1189 individuals 851) Coleoptera (207 individu-als 148) and Hymenoptera (2 individuals 01)Themostcommon families Calliphoridae (987 individuals 706)and Sarcophagidae (102 individuals 73) occupied 779(Figure 3) Lucilia sericata was collected from 22 of 35autopsies (629) and showed the highest relative frequencyfollowedby Sarcophaga peregrina (7 autopsies 114)Amongbeetles Dermestes maculatus was the most common species(4 autopsies 114) The entomofauna observed in thepresent study is summarized in Table 2

It is well known that the insect fauna changes withgeographic region and climate Studies of the insect faunaassociated with human cadavers are fundamental to forensicentomology In experiments with pig carcasses it is difficultto collect insects from various regions unless the experimentsare performed in many different locations However becausethe locations of unusual deaths are varied samples from avariety of regions are included in the present study As theresearch of Caine et al in Portugal [5] the present studyutilized molecular barcoding as well as morphological iden-tification [14ndash21] to ensure confident species identificationThe present study showed a strong preference of incidenceto summer whereas a previous study in Thailand belongingto tropical rainforest did not display any seasonal preference[23]

The dominant species in the present study Lucilia seri-cata showed a strong preference for indoor environmentswhich has been demonstrated in previous studies [24]Calliphora vicina and Calliphora lata showed bimodal sea-sonal occurrence in spring and autumn According to aprevious work by Park in a more southern part of Koreathese cryophilic species tend to prefer cool seasons or highaltitude in summer [25] C vicina a common species inthe Holarctic region is particularly common in Northern

Europe [26] Unlike C vicina the geographic distributionof C lata is known to be limited in the far-east Asia[27] Sarcophagidae species only occurred during summerInterestingly Muscidae species occurred only once in thepresent study Hydrotaea obscrifrons here recorded for thefirst time in Korea This H obscrifrons maggot was collectedfrom a moderately mummified body abandoned inside a carat an underground parking lot in the downtown A previousstudy showed that flies belonging to the genus Ophyra (=Hydrotaea) mainly appear at later stages during ammoniacalfermentation as shown in the present study [26] Piophilacasei (family Piophilidae) which is a potential indicatorof the advanced stage of decay [17 28] was found fromtwo cadavers showing adipocere Although this synanthropicspecieswas expected to exist inKorea thiswas the first officialidentification of this species in Korea Another Piophilidaespecies Parapiophila vulgaris was found to coexist withP casei in one case Maggots of Hermetia illucens (familyStratiomyidae) which are known to feed on a wide varietyof decaying organic materials [29] were found in a severelyputrefied body abandoned in a roomMegaselia scalaris (fam-ily Phoridae) pupae were found in a closed room Becausespecies belonging to the family Phoridae are small in size andcan easily enter closed spaces [30] these flies are potentialindicators for mPMI estimation in cases where other flieshave poor access to the body due to its presence in a closedenvironment [30] Coleoptera species appeared entirely in thesummer except for an individual Dermestes haemorrhoidalisthat was found in a greenhouse in December (Table 2)In Sukontason et alrsquos study in Thailand D maculatus thedominant beetle species in the present study was the onlybeetle species collected [23] Dermestes spp are the firstvisitors of Megninrsquos third wave [31] Silphidae species wereobserved 4 times exclusively in the forest cases Necrodeslittoralis (subfamily Silphinae) andNicrophorus quadraticollis


Table 2 Numbers of medicolegal autopsy cases where each insect species occurred

Scientific names Location Months Total Frs ()I O F Apr May Jun Jul Aug Sep Oct Nov Dec

Order DipteraFamily CalliphoridaeLucilia sericata (Meigen 1826) 21 1 2 1 2 8 4 2 1 1 1 22 629L caesar (Linnaeus 1758) 1 1 1 29L illustris (Meigen 1826) 1 1 1 29Calliphora lata (Coquillett 1898) 2 1 1 2 57C vicina Robineau-Desvoidy 1830 2 1 1 2 57Phormia regina (Meigen 1826) 4 2 1 1 3 2 6 171Chrysomya megacephala (Fabricius 1794) 2 2 2 2 57C pinguis (Walker 1858) 2 2 2 1 1 4 114

Family SarcophagidaeSarcophaga duxThomson 1869 1 1 1 29S crassipalpisMacquart 1839 2 1 1 2 57S peregrina Robineau-Desvoidy 1830 4 3 2 2 3 7 200

Family MuscidaeHydrotaea obscrifrons (Sabrosky 1949) 1 1 1 29

Family PiophilidaePiophila casei (Linnaeus 1758) 1 1 1 1 2 57Parapiophila vulgaris (Fallen 1820) 1 1 1 29

Family StratiomyidaeHermetia illucens (Linnaeus 1758) 1 1 1 29

Family PhoridaeMegaselia scalaris (Loew 1866) 1 1 1 29

Order ColeopteraFamily DermestidaeDermestes ater (DeGeer 1774) 1 1 1 29D frischi Kugelann 1792 1 1 1 1 2 57D haemorrhoidalis Kuster 1852 2 1 2 1 3 86D maculatus (DeGeer 1774) 3 1 2 2 4 114D tessellatocollisMotschulsky 1860 1 1 1 29

Family SilphidaeNecrodes littoralis (Linnaeus 1758) 1 1 1 29Necrodes sp 2 1 1 2 57Nicrophorus quadraticollis Portevin 1903 1 1 1 29

Family HisteridaeSaprinus sp 1 1 1 1 29Saprinus sp 2 1 1 1 29

Family StaphylinidaePhilonthus cyanipennis (Fabricius 1793) 1 1 1 29P longicornis Stephens 1832 1 1 1 29Philonthus sp 1 1 1 29

Family NitidulidaeOmosita sp 1 1 1 29

Family CleridaeNecrobia rufipes (DeGeer 1775) 2 1 1 2 3 86

Order HymenopteraFamily PteromalidaeNasonia vitripennis (Walker 1836) 1 1 1 29

Number of collected species 21 8 13 2 3 9 18 15 3 2 2 2 32I O and F stand for indoor outdoor and forest respectively ldquoFrsrdquo means frequencies


(subfamily Nicrophorinae) were identified Although somespecies in the family Silphidae are known to be remarkableindicators during fresh and bloating stages of decomposition[1] in the present study N quadraticollis was collected fromthe skull of a skeletonized body Only eight individuals werecollected for families Histeridae Staphylinidae and Cleridaeknown to feed on maggots [31] Because the sample sizeof the present study was limited more extensive samplingis required to characterize the forensically important beetlefauna in Korea

In conclusion although all insect species expected to beobserved during a medicolegal autopsy could not be fullycovered due to the limited scale the present study providesa general overview of forensically important insect speciesin Korea Further studies over wider geographical regions inKorea are required to establish more faithful data regardingforensically important entomofauna

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This research was supported by Basic Science ResearchProgram through theNational Research Foundation of Korea(NRF) funded by the Ministry of Education Science andTechnology (2013R1A1A1012223) The authors would like toextend their appreciation to Dr Mitsuhiro Iwasa Laboratoryof Entomology ObihiroUniversity of Agriculture andVeteri-nary Medicine Obihiro Hokkaido Japan for identificationof Piophila casei

References

[1] D E Gennard Forensic Entomology An Introduction JohnWiley amp Sons Chichester UK 2007

[2] P J Saukko and B Knight Knightrsquos Forensic Pathology ArnoldLondonUK 3rd edition 2004 Distributed in theUS byOxfordUniversity Press

[3] H L Lee ldquoRecovery of forensically important entomologicalspecimens from human cadavers in Malaysiamdashan updaterdquoTheMalaysian Journal of Pathology vol 11 pp 33ndash36 1989

[4] M BarretoM E Burbano and P Barreto ldquoFlies (CalliphoridaeMuscidae) and beetles (Silphidae) from human cadavers in CaliColombiardquo Memorias do Instituto Oswaldo Cruz vol 97 no 1pp 137ndash138 2002

[5] L M Caine F C Real M I Salona-Bordas et al ldquoDNA typingof Diptera collected from human corpses in Portugalrdquo ForensicScience International vol 184 no 1ndash3 pp e21ndashe23 2009

[6] Y-H Kim S E Shin C S Ham et al ldquoMolecular identificationof Necrophagous Muscidae and Sarcophagidae fly species col-lected in Korea bymitochondrial cytochrome c oxidase subunitI nucleotide sequencesrdquo The Scientific World Journal vol 2014Article ID 275085 9 pages 2014

[7] S H Park C H Park Y Zhang et al ldquoUsing the develop-mental gene bicoid to identify species of forensically importantblowflies (Diptera Calliphoridae)rdquo BioMed Research Interna-tional vol 2013 Article ID 538051 8 pages 2013

[8] S H Park Y Zhang H Piao et al ldquoUse of cytochrome coxidase subunit I (COI) nucleotide sequences for identificationof the Korean Luciliinae Fly species (Diptera Calliphoridae) inforensic investigationsrdquo Journal of Korean Medical Science vol24 no 6 pp 1058ndash1063 2009

[9] S H Park Y Zhang H Piao et al ldquoSequences of thecytochrome C oxidase subunit I (COI) gene are suitable forspecies identification of Korean Calliphorinae flies of forensicimportance (Diptera Calliphoridae)rdquo Journal of Forensic Sci-ences vol 54 no 5 pp 1131ndash1134 2009

[10] K Saigusa M Takamiya and Y Aoki ldquoSpecies identification ofthe forensically important flies in Iwate prefecture Japan basedon mitochondrial cytochrome oxidase gene subunit I (COI)sequencesrdquo Legal Medicine vol 7 no 3 pp 175ndash178 2005

[11] Q-L Liu J-F Cai Y-F Chang et al ldquoIdentification of foren-sically important blow fly species (Diptera Calliphoridae) inChina by mitochondrial cytochrome oxidase I gene differentia-tionrdquo Insect Science vol 18 no 5 pp 554ndash564 2011

[12] Z J O Adams and M J R Hall ldquoMethods used for the killingand preservation of blowfly larvae and their effect on post-mortem larval lengthrdquo Forensic Science International vol 138no 1ndash3 pp 50ndash61 2003

[13] J Amendt C P Campobasso E Gaudry C Reiter HN LeBlanc and M J R Hall ldquoBest practice in forensicentomologymdashstandards and guidelinesrdquo International Journalof Legal Medicine vol 121 no 2 pp 90ndash104 2007

[14] S H Park ldquoStudies on flies in Korea II Taxonomical studies onsarcophagid flies (Diptera)rdquo Bulletin of the Tokyo Medical andDental University vol 24 no 4 pp 249ndash284 1977

[15] S H Park ldquoStudies on flies in Korea I Taxonomical studies oncalliphorid flies (Diptera)rdquoTheBulletin of the TokyoMedical andDental University vol 24 no 3 pp 189ndash208 1977

[16] S H Park ldquoStudies on flies in Korea III Taxonomic studies onmuscid flies (Diptera Muscidae)rdquoTheBulletin of TokyoMedicaland Dental University vol 25 no 1 pp 17ndash43 1978

[17] M Iwasa ldquoThe piophilidae (Diptera) of Japanrdquo Medical Ento-mology and Zoology vol 49 no 1 pp 33ndash39 1998

[18] BritishMuseum (Natural History) Department of Entomologyand H E Hinton A Monograph of the Beetles Associated withStored Products Trustees of the British Museum London UK1945

[19] Y Kurosawa S Hisamatsu and H Sasaji The Coleoptera ofJapan in Color Volume III Hoikusha Osaka Japan 1985

[20] J I Kim and E J Lim ldquoFauna of Korean histeridae (Coleop-tera)rdquo Entomological Research Bulletin vol 23 pp 59ndash76 1997

[21] Y B Cho and K J Ahn Economic Insects of Korea 11 SilphidaeStaphylinidae (Coleoptera) CIS Seoul Republic of Korea 2001

[22] S Kumar M Nei J Dudley and K Tamura ldquoMEGA abiologist-centric software for evolutionary analysis of DNA andprotein sequencesrdquo Briefings in Bioinformatics vol 9 no 4 pp299ndash306 2008

[23] K Sukontason P Narongchai C Kanchai et al ldquoForensicentomology cases in Thailand a review of cases from 2000 to2006rdquo Parasitology Research vol 101 no 5 pp 1417ndash1423 2007

[24] J H Byrd and J L Castner Forensic Entomology The Utility ofArthropods in Legal Investigations CRC Press Taylor amp FrancisBoca Raton Fla USA 2nd edition 2010

[25] S H Park ldquoThe muscidae calliphoridae and sarcophagidaerdquoin Illustrated Encyclopedia of Diptera of Korea C W Kim SH Park G Y Lee and T J Lee Eds pp 949ndash1042 Sam HwaPublishing Seoul Republic of Korea 1971


[26] K G V Smith ldquoDipterardquo in A Manual of Forensic EntomologyK G V Smith Ed pp 68ndash137 Trustees of the British Museum(Natural History) London UK 1986

[27] R Kano and S Shinonaga Calliphoridae (Insecta Diptera)Biological [ie Biogeographical] Society of Japan Tokyo Japan1st edition 1968

[28] D Martın-Vega ldquoSkipping clues forensic importance of thefamily Piophilidae (Diptera)rdquo Forensic Science Internationalvol 212 no 1ndash3 pp 1ndash5 2011

[29] A Martınez-Sanchez C Magana M Salona and S Rojo ldquoFirstrecord of Hermetia illucens (Diptera Stratiomyidae) on humancorpses in Iberian Peninsulardquo Forensic Science Internationalvol 206 no 1ndash3 pp e76ndashe78 2011

[30] S Reibe and B Madea ldquoUse of Megaselia scalaris (DipteraPhoridae) for post-mortem interval estimation indoorsrdquo Par-asitology Research vol 106 no 3 pp 637ndash640 2010

[31] K G V Smith ldquoColeopterardquo in A Manual of Forensic Entomol-ogy K G V Smith Ed pp 138ndash149 Trustees of the BritishMuseum (Natural History) London UK 1986

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


Table 1 Primer sequences used for the amplification and sequencing of dipteran COI genes

Name Sequence Binding siteF1 51015840-CCTTTAGAATTGCAGTCTAATGTCA-31015840 tRNA-cysteineF2 51015840-GGAGGATTTGGAAATTGATTAGTTCC-31015840 220ndash245 on COIF3 51015840-CTGCTACTTTATGAGCTTTAGG-31015840 1000ndash1022 on COIR1 51015840-CCTAAATTTGCTCATGTTGACA-31015840 2ndash23 on 119862119874119868119868lowast

R2 51015840-CAAGTTGTGTAAGCATC-31015840 1327ndash1343 on COIR3 51015840-CCAAAGAATCAAAATAAATGTTG-31015840 688ndash710 on COIlowast119862119874119868119868 cytochrome c oxidase subunit II gene

collected were preserved in an 80 ethanol solution at 4∘Cand the other half of the live maggots collected were sent tothe incubation chambers for rearing andmorphological taxo-nomic identification Nondipteran samples were immediatelypreserved in 80 ethanol solution without rearing All thesamples immersed in 80 ethanol solution were immediatelysent to a freezer (minus20∘C) Although the best method of killingand preservation of live maggots is treatment in boiling waterand then transfer to 80 ethanol solution to avoid browningand shrinkage of maggots [12 13] the process of immersionin boiling water was omitted because the purpose of thepreservation was merely DNA barcoding

22 Taxonomic Identification Morphological features ofadult individualswere observed under an SZX10 stereomicro-scope (Olympus Corporation Tokyo Japan) Maggots usedfor rearing were taxonomically identified to the species levelafter emergence [14ndash17] After initial identification at a familylevel on the basis of the shape of the posterior spiracles amaggot sample was used for DNA barcoding All the adultbeetles and beetle larvae were taxonomically identified on thebasis of their morphological features to the species and thefamily level respectively [18ndash21]

23 DNA Barcoding of Maggots DNA barcoding of maggotswas performed utilizing the mitochondrial cytochrome coxidase subunit I gene (COI) Polymerase chain reaction wasperformed mainly to amplify the barcoding regions locatedwithin the upstream third portion of COI Primer sequenceswhichwere adopted fromaprevious study are listed inTable 1[6] After automatic sequencing using an ABI Prism 3720xlgenetic analyzer (Applied Biosystems Foster City CA USA)nucleotide sequences were aligned and compared with pre-existing nucleotide sequences using MEGA6 Software [22]

24 Rearing Conditions for Maggots Because the identifica-tion of maggot species usingmorphologic features is difficultspecimens were reared to the adult stage inside incubationchambers under constant conditions (25∘C relative humidity70) The light and dark ratio of each chamber was set at16 8 Live maggots were placed on 5-6 g of fresh pork liver ina plastic cylinder measuring 95mm in height and diameterwith an adequate amount of sawdust under the pork liver

3 Results and Discussion

Weather records for the study period confirmed that north-eastern Seoul belongs to the temperate climate zone having

00

1000

2000

3000

4000

5000

6000

00

100

200

300

400

Jan

Feb

Mar

Apr

May

Jun Jul

Aug

Sep

Oct

Nov

Dec

Rain

fall

(mm

)

Rainfall (mm)Mean

MaximumMinimum

minus200

minus100

Air

tem

pera

ture

(∘C)

Figure 1 A climograph of mean maximum and minimum airtemperatures (∘C) and rainfall (mm) at Seoul weather station(37∘341015840N 126∘581015840E) in 2010 2011 and 2013 demonstrates hot humidsummers and cold dry winters

0

24

6

8

10

12

14

Jan

Feb

Mar

Apr

May

Jun Jul

Aug

Sep

Oct

Nov

Dec

0002 1

4

13

9

221 1

Num

ber o

f aut

opsy

case

s

Month

Figure 2 Medicolegal autopsy cases showing insect infestation inthe study region in 2010 2011 and 2013 were most frequent in thesummer (June July and August)

humid hot summers and dry cold winters with interveningmoderate springs and falls (Figure 1) Insect infestationof human cadavers was most common (26 autopsies) insummer (June July and August) and was not observedfrom January to March (Figure 2) Among 1398 individualsamples a total of 3 orders 13 families 18 genera and




Piophilidae (89) 64


Silphidae (89) 64


Pteromalidae (2) 01

Nitidulidae (1) 01




























Acknowledgments


References








































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




Piophilidae (89) 64


Silphidae (89) 64


Pteromalidae (2) 01

Nitidulidae (1) 01




























Acknowledgments


References








































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology























Acknowledgments


References








































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

research article the first survey of forensically...

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