Research Article Comparative Study of Dipteran Species ...downloads.hindawi.com/journals/insects/2013/398159.pdf · and Their Succession on Rabbit Carrion in Two Different Mangrove

Hindawi Publishing CorporationJournal of InsectsVolume 2013 Article ID 398159 9 pageshttpdxdoiorg1011552013398159

Research ArticleComparative Study of Dipteran Species Diversityand Their Succession on Rabbit Carrion in Two DifferentMangrove Areas of Peninsular Malaysia

Wahizatul Afzan Azmi and S P Lim

Department of Biological Sciences Faculty of Science amp Technology Universiti Malaysia Terengganu21030 Kuala Terengganu Terengganu Malaysia

Correspondence should be addressed to Wahizatul Afzan Azmi wahizatulumtedumy

Received 24 April 2013 Revised 17 July 2013 Accepted 17 July 2013

Academic Editor Francisco de Sousa Ramalho

Copyright copy 2013 W A Azmi and S P Lim This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

A study on dipteran utility in assisting investigation of unattended deaths was carried out in mangrove areas of Kuala TerengganuTerengganu andMasai Johor in Peninsular Malaysia by using rabbit carrions as the modelThe aim of this study was to determinethe dipteran species diversity and their succession over the decomposition period of the rabbit carrions A total of 229 individualsbelonging to 11 species from six families of Diptera were successfully identified fromboth study sites inOctober andDecember 2007Chrysomya megacephala C rufifacies and Hydrotaea sp were found to be the most abundant species recorded in this study Morespecies were collected from Masai with 10 species compared to Kuala Terengganu with nine species Ecological indices (ShannonWiener Index Margalef Index and Evenness Index) showed that Masai scored higher diversity richness and evenness values thanKuala Terengganu However Mann-Whitney 119880 test did not show significant difference among the individuals represented at eachstudy site (119875 gt 005) Calliphoridae predominated in the carrion during the fresh bloat and active decay stages of decompositionDipteran development was documented to be meteorologically dependent whereby low temperature and high rainfall inhibit theircolonization Data collected in this study can hopefully serve as the basis for future estimates of the postmortem interval (PMI)particularly in mangrove area of tropical regions

1 Introduction

Flies (Diptera) are among the most omnipresent and widelydistributed insects Their close association with humans hasled them to be recognized as unpleasant and disturbingcreatures and certainly some flies are responsible formillionsof illnesses and deaths among human populations Yet fliesare also among the key components in most ecosystemsand are advantageous in many ways [1] Because many fliesdevelop from earlier to later stages of a life cycle in the bodiesof dead vertebrates and because several species leave behind aheavily hardened puparial case thatmay endure for centuriesflies are the most significant organisms for forensic studyThey are especially valuable in determining the age of corpsefrom duration of a few hours to a few years [2]

Forensic entomology makes use of information derivedfrom either the succession of arthropods on human corpses

or animal carcasses or the temperature-dependent develop-ment of insects (primarily flies) to estimate the time elapsedsince death or postmortem interval (PMI) estimate of thetime between death and corpse discovery [3] It is importantto know how long it takes for the insect to arrive on a corpsethe stage of decay to which it is attracted its life cycle andits rate of development [4] When a body is found or a crimescene is investigated the presence of insects or lack thereofcan provide many clues as to what actually happened Timeof death can be ascertained andmany clues about the cause ofdeath or events just prior to death can be determined basedon insect behaviour and interaction with a corpse Howeverafter three days insect evidence is often the most accurateand sometimes the only method of determining elapsedtime since death [5] The time of death is a critical piece ofinformation for investigators attempting to understand thecause of suspicious deaths [6]

2 Journal of Insects

In Malaysia most of the studies reported the forensicallyimportant entomological specimens received from hospitalspolice and other agencies throughout Malaysia (eg [78]) Lee and Marzuki [9] studied arthropods succession onmonkey carcasses either fully exposed or partially buriednear the fringes of tropical forest Omar et al [10] madeobservation on arthropods on monkey carrion at a rubbertree plantation in Malaysia However very limited study hasbeen carried out to investigate the diversity of necrophagousinsects in mangrove areasThemangroves forests inMalaysiaare unique ecosystemswhich have a great variety ofmangrovespecies Such areas are commonly subject to extensive sedi-mentation which provides a diverse range of substrate typesand nutrient levels which in turn are favorable for mangrovegrowth [11]Thus diverse of insects such as flies ants beetleswasp mosquitoes and butterflies and other insects occupythis ecosystem as their habitat

Therefore the main objectives of this study were (1) toidentify the dipterans which occurred on the rabbit carrionsin two different mangrove areas which located in the KualaTerengganu Terengganu and in Masai Johor and (2) toinvestigate the dipteran succession pattern as a basis for futureestimates of the PMI The two mangrove areas representingdifferent dominant species of mangroves were selected inorder to investigate the factors that influence the dipterandiversity and succession in different part of peninsularMalay-sia

2 Materials and Methods

21 Study Sites The study was conducted in two differentmangrove areas which were located in Kuala TerengganuTerengganu (05∘241015840358910158401015840N 103∘051015840287110158401015840E) which repre-sents the east coast and in Masai Johor (01∘281015840523010158401015840N103∘531015840066810158401015840E) which represents the southern part ofPeninsular Malaysia Mangroves in Kuala Terengganu aremore exposed to the South China Sea and has more sandysubstrate compared to mudflats in Masai which is moreshady and protected from the waves of sea Mangrove areaat Kuala Terengganu is less developed compared to MasaiMasai is very accessible and is located beside highway roadsconstruction activities are also held nearby the study site Incontrast Kuala Terengganu mangrove area is still in a morepristine condition reserved for research activities and lessperturbed with human activities thus minimizing humaninterference

22 Carrion Four rabbits (Oryctolagus cuniculus L) eachweighing about 13 kg were used and studied for 14 daysin this study Two rabbits were slaughtered and placed inKuala Terengganu mangrove area from 5 to 19 October 2007while another two rabbits were studied in Masai from 8 to22 December 2007 Immediately after death the rabbits wereplaced in round metal cages (40 cm times 20 cm) made with 1 cmwire meshing This allowed access to the carrions by insectswhile prevening disturbances by vertebrate scavengers Thecarrions were photographed dailyTheywere left under shadytrees and the time was recorded Samplings were taken twicea day at 10 am and 5 pm

23 Dipteran Sample Collection Dipterans collected fromdifferent parts of the body were separated into different vialsEach vial and container was labelled with area of the bodyor soil where it was removed date and time of collectionand stage of the insects when collected A duplicate labelwas affixed to the exterior of the vial Adults from the corpsewere collected using a hand net The captured flies were thenkilled immediately by placing them in the killing jar withcotton soaked in 70 of ethanol Upon reaching the labthe adult flies were pinned and the species were determinedSamples of 20ndash35 larvae were collected at the first appearanceof instars and at subsequent of 12 h intervals with jewelerforceps from different areas (such as oral cavities stomachand anus cavities) and placed in glass vials Half of thelarvae samples were killed in hot water and then placed inappropriately labelled bottles containing 70 ethanol [12]These techniques prevent discoloration and shrinkage thatwill occur if living maggots are placed directly into commonpreservative solutions such as ethanol and formaldehydeShrinkage would make the maggots appear younger thanthey actually are These specimens represent the ldquobiologicalclockrdquo to determine the PMI This clock is started with theinvasion of the insects and is stopped by the collection andpreservation of insects from the carrion Each developmentstage of each insect collected from the body represents adistinct interval of time on that clock To determine theinterval represented each sample must be fixed in time anddevelopment by killing and preserving the specimens [13]Another half of the larvae samples were placed on chickenliver and reared to the adult stage for positive taxonomicidentification [12] Labels for the collected samples containedinformation of date time location sample number and adescription of visual observations and notations of the scenebriefly Sampled collections were repeated every 12 hoursuntil the carrion was reduced to a skeleton and no moreapparent insects occurred [14]

24 Identification The identification of the species was donethrough the mounting process of larvae according to thepictorial key by Stojanovish et al [15] and Seago [16]The species was determined by comparing morphologicalfeatures like the presence of an interior apical sklerite shapeand colouration of the structures As for the adult fliesmorphological aspects from the pictorial key according toJason and James [4]Wallman [17] Scott and Borom [18] andDodge [19] were used

25 Postmortem Interval (Succession Pattern) The differentspecies of flies that occurred on the carrion during differentstages of decomposition were identified Fly succession pat-tern in both Kuala Terengganu and Masai were establishedFrom the pattern it gave an indication of the species thatwere present or absent in that location Apart from thatthe succession pattern gave a view of species present duringdifferent stages of decomposition Thus the elapsed timesince death of a dead body was obtained by comparing thedata

Journal of Insects 3

26 Meteorological and Temperature Data Collection Duringsample collection maggot mass temperature was recordedby inserting the thermometer into the center of the maggotmass Soil temperature was taken directly under carrion andat a point 1 to 2m from where the body was laid Dailyweather data that includes precipitation relative humiditydaily minimum and maximum temperatures were obtainedfrom the nearest meteorological station at Sultan MahmudAirport Terengganu and Senai International Airport Johor

27 Data Analysis The degree of species composition orspecies diversity (1198671015840) for each site was determined by usingthe Shannon Wiener Index This index indicates the degreeof species composition per unit area [20]

1198671015840

= minusΣ [(119899119894

119873) ln(119899119894119873)] (1)

where1198671015840 = Shannon-Wiener Index119873 = total individuals ofpopulation sampled and 119899119894 = total individuals belonging tothe 119894 species

Richness Index that has been used was Margalef rsquos Index(119877)This index indicates the number of species in a sample orthe abundance of the species per unit area [20]

119877 =119878 minus 1

ln (119873) (2)

where 119877 = Margalef Richness Index 119878 = total of species and119873 = total of individuals sampled

Homogeneity or pattern of distribution of species inrelation to other species in a sampled per unit area wascalculated using Evenness Index (119864) [20]

119864 =1198671015840

1198671015840max (3)

where 119864 = Evenness Index 1198671015840 = Shannon-Wiener DiversityIndex and1198671015840max=Diversity Index observed to amaximumdiversity

Mann-Whitney 119880 test was used to compare the differ-ences of the total abundance and number of species betweenthe two study sites Spearman correlation analysis was usedto test the relationship of the temperature relative humidityrainfall number of species and abundance

The analyses were conducted using SPSS 160 statisticalsoftware

3 Results and Discussion

31 Dipteran Diversity A total of 229 individuals from11 species of Diptera belonging to six families were col-lected from mangrove areas in Kuala Terengganu and Masai(Table 1) Eight species were commonly found occurred inboth mangrove areas They were Chrysomya megacephalaC rufifacies Sarcophaga sp Musca domestica Homoneuratincta and Cestrotus sp Interestingly two species were foundendemic to mangrove areasHomoneura tincta and Cestrotussp both of them were from family Lauxaniidae Fannia spwas only found in Kuala Terengganu while Phaenicia sericata

Table 1 List of identified dipteran species (both adult and larvae) inKuala Terengganu and Masai

Mangrove(s) Family SpeciesKualaTerengganuTerengganu

Fannidae Fannia sp

Masai Johor Calliphoridae Phaenicia sericataMuscidae Limnophora sp

Bothmangroveareas

Calliphoridae Chrysomya megacephalaChrysomya rufifacies

SarcophagidaeMuscidae

Sarcophaga spMusca domesticaHydorotaea sp

Lauxaniidae Homoneura tinctalowast

Cestrotus splowast

Phoridae Megaselia splowastSpecies found only in mangrove areas

and Limnophora sp were only found in Masai The mostdominant species found in both study sites were Chrysomyamegacephala (31 in Kuala Terengganu 22 in Masai) andC rufifacies (27 in Kuala Terengganu 17 in Masai) Thetwo least species found in Kuala Terengganu and Masaiwere Homoneura tincta and Cestrotus sp both from familyLauxaniidae

Basically scores of all ecological indices were higher atMasai than at Kuala Terengganu Based on Shannon WeinerIndex higher diversity of dipteran was scored at Masai with2056 while Kuala Terengganu scored 1890 Similar trendcould be observed for the Margalef Index where the speciesrichness at Masai and Kuala Terengganu was 1900 and 1686respectively Masai demonstrated higher value of EvennessIndex which was 0893 followed by Kuala Terengganu with0860

However there were no significant differences for num-ber of species (119911 = minus0817 119875 = 0414) number of family(119911 = minus1200 119875 = 0230) and number of individuals (119911 =minus0487 119875 = 0626) between both study sites Based onSpearman correlation analysis mean ambient temperaturesignificantly influenced the dipteran populations both inKuala Terengganu and Masai (119903 = 0488 119875 = 0008) Rainfallstrongly influenced the number of species (119903 = 0410 119875 =0030) and number of individuals (119903 = minus0388 119875 = 0041)in both study sites However no correlation between relativehumidity and dipteran diversity in both sites was detected

32 Succession Pattern Based on Stages of DecompositionFive different stages of decomposition were observed duringthe study fresh bloated active decay advanced decay andskeletal which were determined by the physical weight andbody temperature changes of the carrion Tables 2 and 3show the succession patterns of dipteran species in KualaTerengganu Terengganu andMasai Johor whereas Figures 1and 2 show the decomposition stages for the carrions in bothstudy areas


Table 2 Successional patterns of dipteran species in Kuala Tereng-ganu mangrove area

Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12

Decomposition stagesF BL ACD ADD DR

Chrysomyamegacephala lowast lowast lowast lowast

Chrysomyarufifacies lowast lowast lowast lowast lowast

Sarcophaga sp lowast lowast lowast

Hydrotaea sp lowast lowast lowast lowast lowast

Muscadomestica

Homoneuratincta

Cestrotus sp Megaselia sp lowast lowast lowast lowast lowast lowast

Fannia sp Note F Fresh BL Bloated ACD Active decay stage ADD Advanced decaystage DR Dry remainlowast Adult and immature stages present Only adult individual present

Table 3 Successional patterns of dipteran species in Masai man-grove area

Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12


Chrysomyamegacephala lowast lowast lowast lowast lowast lowast

Chrysomyarufifacies lowast lowast lowast lowast

Phaeniciasericata

Sarcophaga sp lowast lowast

Hydrotaea sp lowast lowast lowast lowast

Muscadomestica

Limnophora sp lowast lowast lowast

Cestrotus sp Homoneuratincta

Megaselia sp lowast lowast lowast lowast lowast lowast

Note F Fresh BL Bloated ACD Active decay stage ADD Advanced decaystage DR Dry remainlowast Adult and immature stages present Only adult individual present

321 Fresh Stage The first species coming to colonize car-rion at Kuala Terengganu was a group of ants identifiedas Oecophylla smaragdina which belongs to FormicidaeThey invaded the neckrsquos wound and the natural orifices of

the body the eyes mouth nose anus and genital They werefound actively fed on fluids from the body and also on adultflies by capturing them whilst the adult flies try to accessthe carrion They also removed maggots from carrion at thelater stages Only a few of adults Calliphoridae which wereC megacephala were observed flying all over the cages andcarcasses and in the surrounding grassy area In contrast thepioneer species visiting the carrion in Masai was Phaeniciasericata By noon ovipositing adults of Phaenicia sericatawere found attracted to the suppurating neck wound and tothe natural opening of the carrion such as nose and eyesLimnophora sp belonging to Muscidae was also observedcolonized the carrion at Masai during the fresh stage

322 Bloated Stage Numerous adult flies were beingobserved to colonize the carrion At Masai Limnophora spfound to be frequent the carrion as well as other blowfliesThese small size species were found harbouring around thenatural cavity such as nose and the mouth of the carrionA few Sarcophaga sp was seen to fly around the cages andgrassy areas for Kuala Terengganu and Masai Chrysomyamegacephala and C rufifacies were found colonized at thisstage at both study sites A few adult species Homoneuratincta was observed at this stage At Kuala TerengganuCalliphoridae laid their eggs at the other body part especiallythe genitals and under the fur of the body caused by theinvasion of the ants around the woundHydrotaea spMuscadomestica and Fannia sp began to visit the carrion at the endof this stage Overall the bloated stage lasted until day 3 forthese study areas

323 Active and Advanced Decay Stage Adult Calliphoridaewere abundant and the carcasses were literally bubblingwith maggot activity Hundreds of first second and thirdinstar maggots shrouded each rabbit carcass As the resultof maggot feeding the head area of many of the carcasseshaddeteriorated leaving detached ears AtKualaTerengganuspeciesHydrotaea spMegaselia sp and Fannia sp colonizedat this stage Whereas at Masai C megacephala Phaeniciasericata Hydrotaea sp and Megaselia sp were observedfrequent the carrion at this stage Less number of Cestrotussp were spotted at this stage at both study sitesThe carcass inKuala Terengganu seemed to active decay more gradually tillday 4 whereas this stage continued till day 5 for Masai After6 to 9 days of the rabbitrsquos death in both sites the weight ofthe carrion dropped to 419 Besides the absence of odourand the removal of a great part of the soft tissues no larvawas observed In contrast pupariumwas seen underneath thecarrion and in between the carrionrsquos bones

324 Skeletal Stage On day 10 at both sites only remainsof dried skin and bones were left which characterized theskeletal stageThe carrionrsquos weight decreased consistently butnot much throughout the skeletal stage where by the endof this study on day 14 128 was left of the carrion Smallnumbers of larvae were found concentrated underneath thecarrion and where small amounts of soft tissues were left in


(a) (b)

(c) (d)

(e) (f)

Figure 1 Decomposition stages of rabbit carrion inmangrove area of Kuala Terengganu Terengganu (Note (a) fresh stage (b) bloated stage(c) active decay stage ((d) and (e)) advanced decay stage (f) skeletal stage)

the stomach No adult flies were observed in this stage Bothcarrions in both sites reached the skeletal stage by day 10

In present study all species identified have been com-monly documented to be forensically important except forHomoneura tincta andCestrotus sp from family LauxaniidaeHomoneura tincta is small robust fly with orange colouringwhereas Cestrotus sp is black colouring with the wings beingopaque and spotted These lauxaniid flies often occur in themangrove swamps [21]

Among all the meteorological factors ambient temper-ature and rainfall were identified to be the strongest factorsaffecting dipteran succession and developmentThe durationof the decay process depended on climatic conditions Thebiomass loss occurred very fast during the fresh to activedecay stages (from day 1 to 6) at Kuala Terengganu mangrove

area while the biomass loss occurred moderately from thefresh to active decay stages (from day 1 to 5) at Masaimangrove area Only approximately 20 of total biomassremained at Kuala Terengganu but the biomass loss of thecarrion atMasai still remained approximately 50 (Figure 3)Carrion in Masai with lower temperature and rainy seasonretarded insect succession and prolonged the decompositionstages Carrion in Kuala Terengganuwith higher temperaturespeeded up succession and decayed at faster rate The highertemperature of the corpse due to maggot activity also leads toquick decay

The rate of decomposition is exceptionally faster thanthat of other study which took a minimum of 30 days ormore to reach the same stageThis situationmay be explainedby the particular conditions of warmer temperatures in this


(a) (b)

(c) (d)

(e) (f)

Figure 2 Decomposition stages of rabbit carrion in mangrove area of Masai Johor (Note (a) fresh stage (b) bloated stage (c) active decaystage ((d) and (e)) advanced decay stage (f) skeletal stage)

study (mean temperatures of 275∘C in Masai 282∘C inKuala Terengganu) compared with the other reports (meantemperatures of 180∘C to 240∘C) In Kuala Terengganuwarmer temperatures and fever precipitation speeded up thesuccession by accelerating the development and activity ofdipterous larvae which in turn produced faster degradationof corpse Payne [22] found that high temperatures duringwarmer days intensified insect activity resulting in a rapiddepletion of the carrion In Masai the low temperaturesretarded the development and activity of dipterous larvaehence speed of succession was slowed as was degradationof corpse The environment is usually the main factor indetermining the postmortem interval since the ambienttemperature has a direct influence on both the putrefaction

processes and insect activity Cold temperatures slow downorganic matter decay inhibiting bacterial proliferation andpreserving corpse tissues longer Warm temperatures speedup the decomposition process increasing bacterial prolifera-tion and also the number and type of carrion insects [23]

In addition while at the active decay decompositionstages there were heavy rainfalls which caused the formationof grayish adipocere (grave wax) at the abdomen Rainsinhibit the decomposition Blow flies are typically sensitive tomoisture levels When moisture is too high blow fly larvaewill leave a carcass and have been noted to cease larvaldevelopment [22] This was prolonged the decompositionstages of the carrion [5] The oviposition of species withinthe insect order Diptera (flies) is negligible at temperatures


120

100

80

60

40

20

00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Fres

h

Bloa

ted

Active decay

Advance decay Dry

remain

Biom

ass r

emai

ning

()

PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

824 824825 803 834 866 776 852 833 813 818 833 835 79981531 31 31 31 31 3231 31 31 31313030 3029

25ndash 25ndash 25ndash 25ndash 25ndash 25ndash 25ndash 25ndash25ndash23ndash 23ndash 24ndash 24ndash 24ndash24ndash

(a)

120

100

80

60

40

20

0

Biom

ass r

emai

ning

()

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

32 32 32 3231 3133 33 33 30 3026 27 24 28

Fres

h

Bloa

ted

Activ

e dec

ay

Advance decay Dry

remain

882 846 857 877 848 864 910 886 928 961 951 971 897 934 885

22ndash 23ndash 23ndash 24ndash 24ndash 24ndash 24ndash 21ndash24ndash24ndash23ndash 24ndash 24ndash22ndash 24ndash

(b)

Figure 3 Percentage of biomass loss of the carrion with the postmortem interval (PMI) in relation to ambient mean temperature humidityand rainfall at (a) Kuala Terengganu and (b) Masai mangrove areas

below 10∘CThis is due to the poikilothermic (cold-blooded)nature of insects Therefore it can be inferred that thedecomposition process will also be considerably slower inthe monsoon season due to the lower average temperaturesThe developmental speed of the resulting larvae was also

decreased due to the low temperatures again lengthening thedecomposition time [14]

Some insects shun light others prefer it it thus followsthat deep shade will affect the species of insects on a car-rion Chrysomya (Diptera Calliphoridae) preferred shady


conditions while Phenicia (Diptera Calliphoridae) and Sar-cophaga (Diptera Sarcophagidae) prefer sunlight Appar-ently shaded area of both sites were mostly preferred andattracted by Calliphoridae This was true when we observedthe carrions in this two study sites with the shady conditionof the mangrove areas contributed by dense canopy of treesattracting a lot of Chrysomya Phaenicia sericata although itvisits the carrion at Masai showed less number Accordingto Cragg [24] the blowfly Phaenicia sericata does not usuallyoviposit on carcasses with a surface temperature below 30∘CTherefore if eggs of this species are found on a human corpselying in a place that is in shadow during the entire day thefinding may be interpreted as indicating that the corpse hasbeen removed from an area in which there was sunshineearlier [25]

Besides meteorological factors smaller carrion biomassavailable will induce shorter postmortem interval or fasterdecomposition rates Here rabbits weighing around onekilogram were used compared with heavier carrions in otherstudies Micozzi [26] found that the composition of insectcommunity did not show significant differences betweenhuman and pig but the amount of carrion available stronglyinfluenced the postmortem interval

This study shows that diversity of species type of speciesnumber of individuals life stages present and the number ofindividuals of each life stage can all be derived from succes-sion studies in different localities Information gained fromthis study is hoped to provide the knowledge of local carrionfauna which provides a complimentary approach to post-mortem estimation particularly in mangrove area of tropicalregions

Conflict of Interests

The authors declare that there is no conflict of interests withany commercial identities mentioned in this paper

Acknowledgments

The authors would like to thank Mr Muhammad binEmbong for assistance during field work and in the labo-ratory and the Department of Biological Sciences Facultyof Science amp Technology Universiti Malaysia Terengganu forthe financial support for this study

References

[1] J H Skevington and P T Dang ldquoExploring the diversity of flies(Diptera)rdquo Biodiversity vol 3 no 4 pp 20ndash21 2002

[2] K L Tabor R D Fell and C C Brewster ldquoInsect fauna visitingcarrion in Southwest Virginiardquo Forensic Science Internationalvol 150 no 1 pp 73ndash80 2005

[3] R D Hall ldquoIntroduction perceptions and status of forensicentomologyrdquo in Forensic Entomology The Utility of Arthropodsin Legal Investigations J H Byrd and J L Castner Eds pp 1ndash15CRC Press Boca Raton Fla USA 2001

[4] H B Jason and L C James Forensic Entomology The Utilityof Arthropods in Legal Investigations CRC Press London UK2001

[5] G S Anderson ldquoMinimum and maximum development ratesof some forensically important Calliphoridae (Diptera)rdquo Jour-nal of Forensic Sciences vol 45 no 4 pp 824ndash832 2000

[6] E P Catts and M L Goff ldquoForensic entomology in criminalinvestigationsrdquo Annual Review of Entomology vol 37 no 1 pp253ndash272 1992

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

[8] H L Lee ldquoForensically important fly maggots recovered fromhuman cadavers in Malaysiardquo Tropical Biomedicine vol 13 pp93ndash94 1996

[9] H L Lee and T M Marzuki ldquoPreliminary observations ofthe occurrence of arthropods on carrion and its application toforensic entomology in Malaysiardquo Tropical Biomedicine vol 10pp 5ndash8 1993

[10] B Omar A M Mohamed S Sulaiman and P OothumanldquoDipteran succession in monkey carion at a rubber tree planta-tion in Malaysiardquo Tropical Biomedicine vol 11 pp 77ndash82 1994

[11] P B Tomlinson The Botany of Mangroves Cambridge Univer-sity Press London UK 1986

[12] L M L Carvalho P J Thyssen M L Goff and A X LinharesldquoObservations on the succession patterns of necrophagousinsects on a pig carcass in an urban area of SoutheasternBrazilrdquoAnil Aggrawalrsquos Internet Journal of ForensicMedicine andToxicology vol 5 no 1 pp 33ndash39 2004

[13] M L Goff A Fly for the Prosecution How Insect Evidence HelpsSolve Crimes HarvardUniversity Press CambridgeMass USA2000

[14] J H Byrd and J L Castner Forensic Entomology The Utility ofArthropods in Legal Investigations CRC Press New York NYUSA 2001

[15] C J Stojanovish H D Pratt and E E Bennington FlyLarvae Key to Some Species of Public Health Importance USDepartment of Health Education and Welfare Atlanta GaUSA 1962

[16] J M Seago Fly Larvae Pictorial Key to Common Species USDepartment of Health Education and Welfare Atlanta GaUSA 1953

[17] J F Wallman ldquoA key to the adults of species of blowflies insouthern Australia known or suspected to breed in carrionrdquoMedical and Veterinary Entomology vol 15 no 4 pp 433ndash4372001

[18] H G Scott and M E Borom Domestic Flies Pictorial Key toCommon Species US Department of Health Education andWelfare Atlanta Ga USA 1962

[19] H R DodgeDiptera Pictorial Key to Principal Families of Pub-lic Health Importance US Department of Health Educationand Welfare Atlanta Ga USA 1953

[20] J L Metcalfe ldquoBiological water quality assessment of runningwaters based on macroinvertebrate communities history andpresent status in Europerdquo Environmental Pollution vol 60 no1-2 pp 101ndash139 1989

[21] L Papp ldquoDipterous guilds of small-sized feeding sources inforests of Hungaryrdquo Acta Zoologica Academiae ScientiarumHungaricae vol 48 supplement 1 pp 197ndash213 2002

[22] J A Payne ldquoA summer carrion study of the baby pig sus scrofaLinnaeusrdquo Ecology vol 46 no 5 pp 592ndash602 1965

[23] C P Campobasso and F Introna ldquoThe forensic entomologist inthe context of the forensic pathologistrsquos rolerdquo Forensic ScienceInternational vol 120 no 1-2 pp 132ndash139 2001


[24] J B Cragg ldquoThe olfactory behaviour of Lucilia species (Diptera)under natural conditionsrdquoAnnals of Applied Biology vol 44 pp467ndash477 1956

[25] B S Shean L Messinger and M Papworth ldquoObservations ofdifferential decomposition on sun exposed v shaded pig carrionin coastalWashington staterdquo Journal of Forensic Sciences vol 38no 4 pp 938ndash949 1993

[26] M S Micozzi ldquoPostmortem change in human and animalremainsrdquo American Journal of Human Biology vol 4 pp 695ndash696 1991

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Enzyme Research



Microbiology


In Malaysia most of the studies reported the forensicallyimportant entomological specimens received from hospitalspolice and other agencies throughout Malaysia (eg [78]) Lee and Marzuki [9] studied arthropods succession onmonkey carcasses either fully exposed or partially buriednear the fringes of tropical forest Omar et al [10] madeobservation on arthropods on monkey carrion at a rubbertree plantation in Malaysia However very limited study hasbeen carried out to investigate the diversity of necrophagousinsects in mangrove areasThemangroves forests inMalaysiaare unique ecosystemswhich have a great variety ofmangrovespecies Such areas are commonly subject to extensive sedi-mentation which provides a diverse range of substrate typesand nutrient levels which in turn are favorable for mangrovegrowth [11]Thus diverse of insects such as flies ants beetleswasp mosquitoes and butterflies and other insects occupythis ecosystem as their habitat

Therefore the main objectives of this study were (1) toidentify the dipterans which occurred on the rabbit carrionsin two different mangrove areas which located in the KualaTerengganu Terengganu and in Masai Johor and (2) toinvestigate the dipteran succession pattern as a basis for futureestimates of the PMI The two mangrove areas representingdifferent dominant species of mangroves were selected inorder to investigate the factors that influence the dipterandiversity and succession in different part of peninsularMalay-sia

2 Materials and Methods

21 Study Sites The study was conducted in two differentmangrove areas which were located in Kuala TerengganuTerengganu (05∘241015840358910158401015840N 103∘051015840287110158401015840E) which repre-sents the east coast and in Masai Johor (01∘281015840523010158401015840N103∘531015840066810158401015840E) which represents the southern part ofPeninsular Malaysia Mangroves in Kuala Terengganu aremore exposed to the South China Sea and has more sandysubstrate compared to mudflats in Masai which is moreshady and protected from the waves of sea Mangrove areaat Kuala Terengganu is less developed compared to MasaiMasai is very accessible and is located beside highway roadsconstruction activities are also held nearby the study site Incontrast Kuala Terengganu mangrove area is still in a morepristine condition reserved for research activities and lessperturbed with human activities thus minimizing humaninterference

22 Carrion Four rabbits (Oryctolagus cuniculus L) eachweighing about 13 kg were used and studied for 14 daysin this study Two rabbits were slaughtered and placed inKuala Terengganu mangrove area from 5 to 19 October 2007while another two rabbits were studied in Masai from 8 to22 December 2007 Immediately after death the rabbits wereplaced in round metal cages (40 cm times 20 cm) made with 1 cmwire meshing This allowed access to the carrions by insectswhile prevening disturbances by vertebrate scavengers Thecarrions were photographed dailyTheywere left under shadytrees and the time was recorded Samplings were taken twicea day at 10 am and 5 pm

23 Dipteran Sample Collection Dipterans collected fromdifferent parts of the body were separated into different vialsEach vial and container was labelled with area of the bodyor soil where it was removed date and time of collectionand stage of the insects when collected A duplicate labelwas affixed to the exterior of the vial Adults from the corpsewere collected using a hand net The captured flies were thenkilled immediately by placing them in the killing jar withcotton soaked in 70 of ethanol Upon reaching the labthe adult flies were pinned and the species were determinedSamples of 20ndash35 larvae were collected at the first appearanceof instars and at subsequent of 12 h intervals with jewelerforceps from different areas (such as oral cavities stomachand anus cavities) and placed in glass vials Half of thelarvae samples were killed in hot water and then placed inappropriately labelled bottles containing 70 ethanol [12]These techniques prevent discoloration and shrinkage thatwill occur if living maggots are placed directly into commonpreservative solutions such as ethanol and formaldehydeShrinkage would make the maggots appear younger thanthey actually are These specimens represent the ldquobiologicalclockrdquo to determine the PMI This clock is started with theinvasion of the insects and is stopped by the collection andpreservation of insects from the carrion Each developmentstage of each insect collected from the body represents adistinct interval of time on that clock To determine theinterval represented each sample must be fixed in time anddevelopment by killing and preserving the specimens [13]Another half of the larvae samples were placed on chickenliver and reared to the adult stage for positive taxonomicidentification [12] Labels for the collected samples containedinformation of date time location sample number and adescription of visual observations and notations of the scenebriefly Sampled collections were repeated every 12 hoursuntil the carrion was reduced to a skeleton and no moreapparent insects occurred [14]

24 Identification The identification of the species was donethrough the mounting process of larvae according to thepictorial key by Stojanovish et al [15] and Seago [16]The species was determined by comparing morphologicalfeatures like the presence of an interior apical sklerite shapeand colouration of the structures As for the adult fliesmorphological aspects from the pictorial key according toJason and James [4]Wallman [17] Scott and Borom [18] andDodge [19] were used

25 Postmortem Interval (Succession Pattern) The differentspecies of flies that occurred on the carrion during differentstages of decomposition were identified Fly succession pat-tern in both Kuala Terengganu and Masai were establishedFrom the pattern it gave an indication of the species thatwere present or absent in that location Apart from thatthe succession pattern gave a view of species present duringdifferent stages of decomposition Thus the elapsed timesince death of a dead body was obtained by comparing thedata




1198671015840

= minusΣ [(119899119894

119873) ln(119899119894119873)] (1)



119877 =119878 minus 1

ln (119873) (2)



119864 =1198671015840

1198671015840max (3)








Fannidae Fannia sp


Bothmangroveareas





Cestrotus splowast








Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12






Muscadomestica

Homoneuratincta




Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12




Phaeniciasericata



Muscadomestica











(a) (b)

(c) (d)

(e) (f)








(a) (b)

(c) (d)

(e) (f)






120

100

80

60

40

20

00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Fres

h

Bloa

ted

Active decay

Advance decay Dry

remain

Biom

ass r

emai

ning

()

PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

824 824825 803 834 866 776 852 833 813 818 833 835 79981531 31 31 31 31 3231 31 31 31313030 3029


(a)

120

100

80

60

40

20

0

Biom

ass r

emai

ning

()

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

32 32 32 3231 3133 33 33 30 3026 27 24 28

Fres

h

Bloa

ted

Activ

e dec

ay

Advance decay Dry

remain

882 846 857 877 848 864 910 886 928 961 951 971 897 934 885


(b)











Acknowledgments


References



































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




1198671015840

= minusΣ [(119899119894

119873) ln(119899119894119873)] (1)



119877 =119878 minus 1

ln (119873) (2)



119864 =1198671015840

1198671015840max (3)








Fannidae Fannia sp


Bothmangroveareas





Cestrotus splowast








Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12






Muscadomestica

Homoneuratincta




Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12




Phaeniciasericata



Muscadomestica











(a) (b)

(c) (d)

(e) (f)








(a) (b)

(c) (d)

(e) (f)






120

100

80

60

40

20

00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Fres

h

Bloa

ted

Active decay

Advance decay Dry

remain

Biom

ass r

emai

ning

()

PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

824 824825 803 834 866 776 852 833 813 818 833 835 79981531 31 31 31 31 3231 31 31 31313030 3029


(a)

120

100

80

60

40

20

0

Biom

ass r

emai

ning

()

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

32 32 32 3231 3133 33 33 30 3026 27 24 28

Fres

h

Bloa

ted

Activ

e dec

ay

Advance decay Dry

remain

882 846 857 877 848 864 910 886 928 961 951 971 897 934 885


(b)











Acknowledgments


References



































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12






Muscadomestica

Homoneuratincta




Diptera species

Day (PMI)1 2 3 4 5 6 7 8 9 10 11 12




Phaeniciasericata



Muscadomestica











(a) (b)

(c) (d)

(e) (f)








(a) (b)

(c) (d)

(e) (f)






120

100

80

60

40

20

00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Fres

h

Bloa

ted

Active decay

Advance decay Dry

remain

Biom

ass r

emai

ning

()

PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

824 824825 803 834 866 776 852 833 813 818 833 835 79981531 31 31 31 31 3231 31 31 31313030 3029


(a)

120

100

80

60

40

20

0

Biom

ass r

emai

ning

()

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

32 32 32 3231 3133 33 33 30 3026 27 24 28

Fres

h

Bloa

ted

Activ

e dec

ay

Advance decay Dry

remain

882 846 857 877 848 864 910 886 928 961 951 971 897 934 885


(b)











Acknowledgments


References



































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


(a) (b)

(c) (d)

(e) (f)








(a) (b)

(c) (d)

(e) (f)






120

100

80

60

40

20

00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Fres

h

Bloa

ted

Active decay

Advance decay Dry

remain

Biom

ass r

emai

ning

()

PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

824 824825 803 834 866 776 852 833 813 818 833 835 79981531 31 31 31 31 3231 31 31 31313030 3029


(a)

120

100

80

60

40

20

0

Biom

ass r

emai

ning

()

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

32 32 32 3231 3133 33 33 30 3026 27 24 28

Fres

h

Bloa

ted

Activ

e dec

ay

Advance decay Dry

remain

882 846 857 877 848 864 910 886 928 961 951 971 897 934 885


(b)











Acknowledgments


References



































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


(a) (b)

(c) (d)

(e) (f)






120

100

80

60

40

20

00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Fres

h

Bloa

ted

Active decay

Advance decay Dry

remain

Biom

ass r

emai

ning

()

PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

824 824825 803 834 866 776 852 833 813 818 833 835 79981531 31 31 31 31 3231 31 31 31313030 3029


(a)

120

100

80

60

40

20

0

Biom

ass r

emai

ning

()

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

32 32 32 3231 3133 33 33 30 3026 27 24 28

Fres

h

Bloa

ted

Activ

e dec

ay

Advance decay Dry

remain

882 846 857 877 848 864 910 886 928 961 951 971 897 934 885


(b)











Acknowledgments


References



































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


120

100

80

60

40

20

00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Fres

h

Bloa

ted

Active decay

Advance decay Dry

remain

Biom

ass r

emai

ning

()

PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

824 824825 803 834 866 776 852 833 813 818 833 835 79981531 31 31 31 31 3231 31 31 31313030 3029


(a)

120

100

80

60

40

20

0

Biom

ass r

emai

ning

()

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PMI (day)

Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Temp (∘C)

RH ()Rainfall (mm) 0 0 0 0 0 0 0 0 0 0002 14 44 34

32 32 32 3231 3133 33 33 30 3026 27 24 28

Fres

h

Bloa

ted

Activ

e dec

ay

Advance decay Dry

remain

882 846 857 877 848 864 910 886 928 961 951 971 897 934 885


(b)











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








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

Research Article Comparative Study of Dipteran Species ...downloads.hindawi.com/journals/insects/2013/398159.pdf · and Their Succession on Rabbit Carrion in Two Different Mangrove