Content uploaded by Abesh Chakraborty
Author content
All content in this area was uploaded by Abesh Chakraborty on Dec 14, 2016
Content may be subject to copyright.
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS)
e-ISSN: 2319-2380, p-ISSN: 2319-2372. Volume 8, Issue 8 Ver. II (Aug. 2015), PP 79-89
www.iosrjournals.org
DOI: 10.9790/2380-08827989 www.iosrjournals.org 79 | Page
Developmental analysis of immature stages of Sarcophaga
(Parasarcophaga) albiceps Meigen,1826 (Diptera:Sarcophagidae)
on Gallus gallus carcass:Their applications as forensic indicators
Chakraborty, A.1*; Ghosh, S.2 ; Ansar,W.3 , Banerjee,D.1
1 Zoological Survey of India, Ministry of Environment & Forests (Government of India), M Block, New Alipore,
Kolkata-700 053, West Bengal,India
2School of Biological Sciences, National Institute of Science, Education and Research (NISER), Institute of
Physics Campus, Sachivalaya Marg, PO: Sainik School, Bhubaneswar - 751 005,Odhisa, India.
3Dept of Zoology, Post Graduate Dept, Asutosh College, 92, S.P. Mookerjee Road Kolkata-700026,
West Bengal, India.
Email: abeshc1@gmail.com
Abstract : The blow flies (Calliphoridae) and flesh flies (Sarcophagidae) are among the first wave of faunal
sucession on human and animal cadavers.Thus, their immature stages are used to estimate the post mortem
interval (PMI).The larval development might vary seasonally.A study was conducted to investigate the effect of
seasonal variation on develoment of immature stages (larvae and pupa) of Sarcophaga (Parasarcophaga)
albiceps on carcass of Gallus gallus (n=3). Results showed no significant diffrences in the average length,
average width and calculated average biomass of the imature stages as [F(2,9)=0.0184, p= 0.9817],
[F(2,9)=0.2415, p= 0.7903] and [F(2,9)=0.5254, p= 0.6083] respectively. These results might have important
implications to forensic entomologists, (p >0.05). Since one approach of PMI estimation uses laravae collected
from crimes scene and compareing them with reference data, derived from rearing of larvae.The results indicate
that Sarcophaga (Parasarcophaga) albiceps can be utilized as forensic indicator for above said pourpose, as
there is no significant developmental variation of immature stages seasonally, when reared on Gallus gallus
carcass.
Keywords: Forensic dipterology, India, Sarcophagidae, Forensic entomology, Sarcophaga (Parasarcophaga)
albiceps, Avian model.
I. Introduction
Nature have provided insects with built in receptors, to assess, quantify and locate caracass of both
human and animals, forensic entomology uses this precarious property of insects namely, the orders Diptera and
Coleoptera, to estimate minimum time since death. Post mortem changes of decaying organisms depend on
various factors, Micozzi (1991). Estimating post mortem interval thus becomes a rather difficult task, as time
since death increases the normal physiological and pathological tests reliability and accuracy decreases, Bass
(1984). Since the normal physiological and pathological tests for estimating PMI yeild ambigious results after
84 hours, Henssage (1995) for both human and non-human carrion.
Therefore to avoid ambiguity in PMI estimates and to obtain more accurate information, other process
are needed to be investigated for establishing correct PMI. It has been found that any physical or biochemical
change that is a function of time since death can be utilized for the aforesaid pourpose, better known as PMI
dependent process. A carcass is a depleting resource, which attracts a variety of scavengers, Putman (1983).
Generally to estimate PMI for vertebrate carcass various process are used, whose principals are based on PMI
dependent process, Hall (1990). One of this process is the immature insect species development model, were
immature stages of the insects consume dead human or animal tissue at diffrent stages of their life cycle. The
estimated age of an immature insects that feeds on the carrion provides the minimum PMI. Investigators in
India, when faced with a need to estimate a portion of the PMI from entomological data from the scene lack this
reference data on this model. The current study aims to bridge this gap, for wildlife crimes pertaining to avian
model and extrapolation of the data can be used for human model, Sarcophaga (Parasarcophaga) albiceps
Meigen,1826 being the forensic indicator and model organism for immature insect species development on
avian caracass.[1]
The family Sarcophagidae has 126 species recorded from India. The adult flies of the largest members
of Indian Sarcophagidae are ♂ Sarcophaga (Parasarcophaga) albiceps (Meigen,1826) their size ranging from 11-
18 mm. S.albiceps have been known cause tissue myiasis in bulls and breed in human and rabbit excretements.
Adults of this species seem to be active throught the year and common in this part of India, making them ideal
candidates for being forensic indicators for avian and human models. Sarcophaga (Parasarcophaga) albiceps
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 80 | Page
(Meigen,1826) is an arthopod, that belongs to class Insecta, order Diptera, family Sarcophagidae and subfamily
Sarcophaginae. The genus Sarcophaga was first described by Meigen and the subgenus Parasarcophaga was
first described by Jhonston and Tiegs. Nandi had re-described the Indian species as Parasarcophaga albiceps
but the present accepted valid name is Sarcophaga (Parasarcophaga) albiceps. [2-8]
This fly is almost cosmopolitan in distribution, it is found in PALAEARCTIC - Albania, Armenia,
Austria, Azerbaijan, Belgium, Bulgaria, Byelorussia, China (Gansu, Hebei, Heilongjiang, Henan, Hubei,
Jiangsu, Jilin, Liaoning, Neimenggu, Ningxia, Shaanxi, Shandong, Shanghai, Shanxi, Sichuan, Xizang), Croatia,
Czech Republic, Finland, France, Germany, Greece, Gruzia, Hungary, Israel, Italy, Japan (Hokkaido, Honshu,
Kyushu, Shikoku), Kazakhstan, Latvia, Moldova, Netherlands, North Korea, Norway, Poland, Portugal,
Romania, Russia (Central European Territory, East Siberia, Far East, North European Territory, South European
Territory, West Siberia), Serbia, Slovakia, South Korea, Spain, Sweden, Switzerland, Turkey, Ukraine, United
Kingdom. AFROTROPICAL - Kenya. ORIENTAL - Andaman Is, Bangladesh, Bhutan, Indonesia (Flores, Java,
Kalimantan, Lombok, Sulawesi, Sumatra, Timor), China (Fujian, Guangdong, Guangxi, Guizhou, Hainan,
Hunan, Jiangxi, Yunnan, Zhejiang), Japan (Ryukyu Is), Malaysia (West Malaysia), Laccadive Is, Nepal,
Pakistan, Philippines, Singapore, Sri Lanka, Taiwan, Thailand, Vietnam. AUSTRALASIAN/OCEANIAN -
Australia (Queensland), Hawaiian Is (Hawaii, Kauai, Maui, Molokai, Oahu), Indonesia (Irian Jaya, Moluku),
Papua New Guinea (Bismarck Arch.), Solomon Is.[2,3-8]
S.albiceps has been bred from rabbit and human excretamentment.They are found to be Parasitic on
Nonagria sp. And cause tissue myiasis in bulls.The male attracted to Aristolochia sp. (Senior-White,Aubertin
and Smart, 1940). S.albiceps breeds in decaying organic matter and has been observed to larviposit on mutton in
India and fish in Pakistan (Shazia et al., 2006; Singh & Bharti, 2008). Similar observations have been made of
this species breeding on faeces in the presence of carrion in Thailand (Bänziger and Pape 2004). Also, S.
albiceps has been documented causing cutaneous myiasis of buffalo, cows and humans (Castro et al. 2010).
Larvae of S. albiceps are also economically important organisms as they are facultative predators of a variety of
butterfly (Lepidoptera) pupae and Hymenoptera larvae. [9-15]
The base line data generated from this study might also help in stored product entomology, were this flies
can potentially identify rotting meat, which are not suitable for human consumption, biocontrol of Lepidoptera
and Hymenoptera, myiasis biology and also can be used in spread of enteric disease and its prediction model
(Greenberg,1971). The current study only focuses on the morphotaxonomy, life cycle assesment, seasonal
variations and distribution mapping to assess the imature insects development on avian caracass, which might
help PMI estimation for avian caracass in the states where Sarcophaga (Parasarcophaga) albiceps is
distributed.
II. Materials And Methods
II.a .) Study Site
The study was conducted in Kolkata, ZSI, Latitude :22° 30' 51.6888" and Longitude: 88° 19' 30.5256"
were recorded by GPS meter.A dead Gallus gallus (Linnaeus, 1758) was a bought from a market near
Zoological Survey (ZSI) of India (n=3) for three seasons, Kolkata premises, and was kept in ambient outdoor
conditions, inside the ZSI premises. The data for the abiotic factors was gathered from the meteorological data
was collected from the Meterological Department, Alipore, Kolkata. The local climatic regimes of West Bengal
is sub divided into three seasons, viz., pre monsoon (March to June), monsoon (July to October) and post
monsoon (November to February). During the experiments, the measured average temperature (°C ) ranged
from 35 – 43, in the pre monsoon season, 37 – 30, in the monsoon season and 35 – 25, in the post monsoon
season. Relative humidity (%) ranged from 59 – 45, in the pre monsoon season, 90 - 75, during the monsoon
season and 42 - 35, in the post monsoon season. Average precipatation was null during the pre and post
monsoon seasons, the range of the monsoon season was found to be 58 – 35. And average wind speed (km/hr)
ranged from 31 - 13, in the pre monsoon season, 20 - 12, in the monsoon and 15 – 8, in the post monsoon
season. (See table.1).
II.b .) Collection of the fly specimen
The chicken carcass was placed on a raised platform, surrounded by water on all sides to discourage
ants and malise trap was used for overhead capture of dipteran specimens. The malise trap was used to capture
the flies in one part of ZSI,Kolkata, in 2013-2014. As the post feed maggots (larvae of 3rd instar) starts their
migration away from the carcass and into suitable pupation material, in this case a cotton was used to help the
larvae to pupate, and then the contents of that peice of cotton, was placed into a glass container, to see
progression of normal development. From here few immature specimens (n = 4) were collected for taxonomic,
LM and ecologic studies.
A modified version of the malise trap was utilized to capture adults, the schematics and collection of
the adult. Diffrent developmental stages were observed on taxonomic and ecologic basis, and the immature
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 81 | Page
insects were collected with the help of fine forceps for taxonomic studies and were killed with ether and
preserved in 70% alcohol. The experiments, taxonomic and morphological identification and quantification
were carried out in the above mentioned Laboratory of Diptera Section, Kolkata, India. The meteorological data
was collected from the Meterological Department, Alipore, Kolkata.
The construct was checked regularly and notes on the apperance time of S.albiceps, their larvipositing
time, larval developmental time, pupal developmental to adult emergence, were photodocumented by the nikkon
p520 camera and Leica EZ4 HD.
II.c .) Morphological identification of the different developmental stages of Sarcophaga (P) albiceps
The morphology of Sarcophaga (P) albiceps was observed under light microscope (LM) at each stage
in its life cycle. To observe the anatomical feature of the genitalia, both male and female flies were dissected
and examined under LM [18].The male genitalia and the 5th sternite, are the most important characteristic
feature used to differentiate different flesh fly species. The abdominal segments between 3rd and 4th segments
of the flies were dissected on the clean glass slide using a sharp blade, transferred to a mixture of 10% sodium
hydroxide and 95% ethanol for 3 days and following procedures were done as detailed in ref.14, 18. The
genitalia and ovipositor were observed under Leica M205 Stereozoom dissecting microscope and photographs
taken by the allied Leica camera.The growth and development of Sarcophaga (P) albiceps initiating from the
first instar larvae, to development into second, third instar larvae, pupae, imago and finally emergence of the
adult fly (Figs. 1) on dead carrion of the Gallus gallus (n=3) is being reported for the first time from India thus
establishing the role of Sarcophaga (P) albiceps as a forensic indicator for wild life crime and stored products
entomology from India.
II.d) Developmental data analysis
The immature stages (larvae and pupa) and adults were measured to see the rate of development of Sarcophaga
(P) albiceps in the carcass of Gallus gallus (n=3), in all the seasons. The length, width and biomass of larvae (n
= 5) were calculated from Leica EZ4 HD microscopic measurement data for all three seasons. The time spent
was monitored recorded and the percentage of time spent in each stage was calculated.The effects of various
seasons on the growth and development (length, width and biomass) of S. albiceps were subjected to a one way
ANOVA. (Sokal and Rohlf 1981).
III. Results
III.a) Morphology-based identification
III.a.1) Description of the morphology of the male and female adult fly
III.a.1.1) Male Morpho-description
The males are generally of variable size ranging from moderately large to large (11-17 mm).The width
of the frons about 3/5th to that of an eye. Frontal vittae, para-frontal and para-facial, black with silvery pollen on
it. Antennae darkish brown. The 1st segment of the antennae seems to be blackish brown, 2nd segment darkish
black and 3rd moderately brown with whitish pollen. Vibrissae and arista long and plumose along basal 2/3rd of
ficial ridge.Palpi brownish.Probosics jet black and stout. Thorax greyish in colour with three longitidunal black
stripes. Pro stigmatic and pro pleural bristles well developed and accompanied with short hairs. Pro and
mesothoracic spiracles brown; latero-scutellar, apico-scutellar and disco-scutellar bristles were 3, 1, and 1 pair
each respectively.
Wings hayline, with brownish veins; R1 bare ;R 4+5 with a row of 9 short setae located dorsally and
extending to atleast the half of basal node upto r-m. Short setae long ventral surface of basal node of R 4+5 ; fifth
costal segment a little shorter than the 3rd , the former with short spines along basal half of its anterior margin;
costal spines stout; epaulet black with short spines; basicostal scale brown; squma white; halter brown.
Legs black in colour; fore femur with a pair of rows of long bristles along posterodorsal surface and a
row of bristles along posterior margin of ventral surface. Mid femur with a row of 3 bristles along middle
portion of anterolateral surface; Fore tibia with a row of 2-3 short bristles; hind tibia with 2 bristles on middle
portion of posterodorsal surface.
Abdomen with silvery grey checkered pattern. The 2nd and 3rd sternites each with 4 bristles, 4th and 5th
with 2 long bristles.The 5th sternite Y-shaped with stout spines laterally and long hairs terminally on arms.
The genetial capsule looks oddly golden yellow to brownish yellow in colour. The 1st and 2nd genital
segments reddish-brown in color; inner forceps stout, elongated, without protuberance and spines at inner
surface of subapical part, but with tuft of long hairs along basal half; in contrast the outer forceps elongated,
somewhat kidney shaped with hairs along distal half. As per ref. 2, 4.(see Fig.1)
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 82 | Page
III.a.1.2) Female Morpho-description
The females are generally,shorter than the males (7 - 12 mm).There is little biometric diffrence with the
males. Apico-scutellar bristles were absent in females. Chaetotaxy of legs same as in males.The 2nd and 3rd
sternites each with 4 bristles, 4th and 5th with 2 long bristles, 6th broader than the rest, 7th without hairs but with
a row of stout marginal bristles, 8th bare, membranous and slightly concave; anal sternite with short hairs; 7th
tergite with long bristles. As per ref. 2, 4.( (see Fig.1)
III.a.1.3) Description of the morphology of the developmental stages (larva and pupa) of Sarcophaga (P)
albiceps Morphological features of the immature stages special attention was given to 3rd instar larvae and pupa
of Sarcophaga (P) albiceps, which revealed distinct morphological parameters of larvae.To identify the larval
stages attention was given to the LM of anterior and posterior spiracles and 2nd and 3rd integument of pupa. The
identifying criteria utilized for the identification of Sarcophaga (P) albiceps 3rd instar larvae and pupae was the
presences of spines on the interband area of the thoracic and abdominal segments, which are long and pointed.
(see Fig.2)
III.b.) Ecology and Life cycle analysis
The Sarcophaga (P) albiceps seems to be larviparous and therefore no eggs were found from the
carcass in all the three seasons. Sarcophaga (P) albiceps completes its life cycle in 326.11 hrs in premonsoon,
336.38 hrs in monsoon and 343.09 hrs in post monsoon season, the weight of the three carcass being equal for
all the seasons. Therefore on average Sarcophaga (P) albiceps tends to complete its life cycle in Gallus gallus
carcass in about 335.43 hrs.We observed the seasonal variation of diffrent phases of life cycle of Sarcophaga
(P) albiceps, and plotted in bar charts (see Table.2 and Fig.3). Distribution map was created to assess where this
data may be utilised in future, this is as per the distribution of Sarcophaga (P) albiceps, as per liturature and
other collection data sources. (see Fig.4)
III.c.) Developmental data analysis
The data of the LM measurements were utilized for analysis of the diffrent developmental stages and
adult. The time spent in various stages are averaged and standard error was calculated for the same. The length
and width of the various stages were also averaged and standard error was calculated for the same. The whole
process was done for all the seasons. The biomass was calculated from the primary LM data of length and
width, and putting them in fourmula for cylinder, biomass was calculated, afterwards this data was averaged and
standard error was calculated. The calculated biomass might fit the immature stages better than the adult stages,
since the legs and wings arent accounted for in this calculations method. ANOVA (1-way) was done for all the
stages averages of (length, width and calculated biomass) for all the seasons.
IV. Discussion
IV.a.) Developmental data analysis (Seasonal)
Each developmental stages,were stopped wacthed and the averages were calculated.The study was
conducted in 2014. The female Sarcophaga (Parasarcophaga) albiceps, generally arrives at a carcass after
around 24 hours. After their mating with the males, females larviposits.
IV.a.1.) Premonsoon season (PRM)
From initiation of larviposition to deposition of 1st instar larvae it took around 27 hours and 16 minutes,
from 1st instar larvae to 2nd instar larvae it took around 31 hours and 40 minutes, from 2nd instar larvae to 3rd
instar larvae it took around 46 hours and 37 minutes, from 3rd instar larvae to pupae it took around 111 hours
and 8 minutes and from pupae to adult it took around 110 hours and 10 minutes.Therefore the total life cycle
took around 326 hours and 11 minutes to complete its life cycle. (See Graph.1 and Graph.2.)
IV.a.2.) Monsoon season (M)
From initiation of larviposition to deposition of 1st instar larvae it took around 28 hours, from 1st instar
larvae to 2nd instar larvae it took around 32 hours and 30 minutes, from 2nd instar larvae to 3rd instar larvae it
took around 47 hours and 38 minutes, from 3rd instar larvae to pupae it took around 115 hours and 23 minutes
and from pupae to adult it took around 113 hours and 47 minutes.Therefore the total life cycle took around 336
hours and 38 minutes to complete its life cycle. (See Graph.1 and Graph.2.)
IV.a.3.) Postmonsoon season (PSM)
From initiation of larviposition to deposition of 1st instar larvae it took around 28 hours and 50 minutes,
from 1st instar larvae to 2nd instar larvae it took around 33 hours and 28 minutes, from 2nd instar larvae to 3rd
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 83 | Page
instar larvae it took around 48 hours and 23 minutes, from 3rd instar larvae to pupae it took around 117 hours
and 20 minutes and from pupae to adult it took around 115 hours and 48 minutes.Therefore the total life cycle
took around 343 hours and 9 minutes to complete its life cycle. (See Graph.1 and Graph.2.)
IV.a.4.) Average annual developmental time
Therefore on average from initiation of larviposition to deposition of 1st instar larvae it took around 28
hours and 28 minutes, from 1st instar larvae to 2nd instar larvae it took around 32 hours and 32 minutes, from 2nd
instar larvae to 3rd instar larvae it took around 47 hours and 32 minutes, from 3rd instar larvae to pupae it took
around 114 hours and 50 minutes and from pupae to adult it took around 113 hours and 1 minute.Therefore the
total life cycle took around 335 hours and 43 minutes on average to complete its life cycle. With the average
prevalant abiotic conditions. (see Table.1 and Graph.1.)
IV.b.Developmental data analysis (Biometric)
One approach in PMI estimation involves killing the larvae collected from crime scene and comparing
the measured length with reference data derived from rearing of larvae in the laboratory where they are most
commonly fed the liver (Willams and Richardson,1984; Anderson,2000; Grassberg et.al., 2003) or ground beef
(Wells and La Motte,1995) of various mammalian species. In the present study Gallus gallus has been utilized
as bait to attract Sarcophaga (Parasarcophaga) albiceps for three seasons (n = 3). The few of the immature
specimens from stages were seperated from carcass (n = 4) and light microscopy was done for both taxonomic
and biometic studies. The length and width were recorded in mm and the biomass was calculated of immature
specimens were calculated using the fourmula of volume of cylinder from the website
http://www.onlineconversion.com/object_volume_cylinder_tank.htm, the volume was calculated for the
immature stages, and recorded in mm³. The adults were not included in the biometric study as they serve almost
no pourpose in this immature stage biometric study based PMI estimation.Standard deviation (s) and standard
error was calculated for length, width and biomass (Huth.et.al., 1994).
IV.b.1.) Premonsoon season (PRM)
In the premonsoon season the length of the 1st instar larvae was found to be 0.3817 ± 0.0085 (s =
0.0170). The length of the 2nd instar larvae was found to be 1.012 ± 0.1121 (s = 0.2243).The length of the 3rd
instar larvae was found to be 1.8845 ± 0.1513 (s = 0.3026). The length of the pupae was found to be 1.3751 ±
0.0483 (s = 0.0967). The width of the the 1st instar larvae was found to be 0.0775 ± 0.0057 (s = 0.0114). The
width of the 2nd instar larvae was found to be 0.2445 ± 0.0318 (s = 0.0637). The width of the 3rd instar larvae
was found to be 0.5017 ± 0.0360 (s = 0. 0712).The width of the pupae was found to be 0.5242 ± 0.0145 (s =
0.0291). The biomass of the 1st instar larvae was found to be 0.0017 ± 0.0002 (s = 0.0004). The biomass of the
2nd instar larvae was found to be 0.0538 ± 0.0154 (s = 0.0309).The biomass of the 3rd instar larvae was found to
be 0.3882 ± 0.0776 (s = 0. 1552).The biomass of the pupae was found to be 0.2985 ± 0.0237 (s = 0.0476). (see
Table.2)
IV.b.2.) Monsoon season (M)
In the premonsoon season the length of the 1st instar larvae was found to be 0.3517 ± 0.0028 (s =
0.0057). The length of the 2nd instar larvae was found to be 0.9130 ± 0.0988 (s = 0.1977).The length of the 3rd
instar larvae was found to be 1.6875 ± 0.0447 (s = 0.0895).The length of the pupae was found to be 1.3750 ±
0.0368 (s = 0.0736). The width of the the 1st instar larvae was found to be 0.0475 ± 0.0237 (s = 0.0057). The
width of the 2nd instar larvae was found to be 0.1950 ± 0.0975 (s = 0.0794). The width of the 3rd instar larvae
was found to be 0.3060 ± 0.0455 (s = 0.0910).The width of the pupae was found to be 0.4115 ± 0.0564 (s =
0.1129). The biomass of the 1st instar larvae was found to be 0.0005 ± 0.0000 (s = 0.0001). The biomass of the
2nd instar larvae was found to be 0.0336 ± 0.0152 (s = 0.0305).The biomass of the 3rd instar larvae was found to
be 0.1353 ± 0.0424 (s = 0.0849).The biomass of the pupae was found to be 0.1621 ± 0.0483 (s = 0.0966). (see
Table.2)
IV.b.3.) Postmonsoon season (PSM)
In the premonsoon season the length of the 1st instar larvae was found to be 0.3720 ± 0.0043 (s =
0.0087). The length of the 2nd instar larvae was found to be 1.0132 ± 0.0706 (s = 0.1413).The length of the 3rd
instar larvae was found to be 1.7450 ± 0.1245 (s = 0.2490).The length of the pupae was found to be 1.3650 ±
0.0500 (s = 0.1001). The width of the the 1st instar larvae was found to be 0.0116 ± 0.0058 (s = 0.0116). The
width of the 2nd instar larvae was found to be 0.2312 ± 0.1156 (s = 0.0680). The width of the 3rd instar larvae
was found to be 0.4892 ± 0.0358 (s = 0.0717).The width of the pupae was found to be 0.5217 ± 0.0140 (s =
0.0280). The biomass of the 1st instar larvae was found to be 0.0017 ± 0.0002 (s = 0.0004). The biomass of the
2nd instar larvae was found to be 0.0473 ± 0.0149 (s = 0.0299).The biomass of the 3rd instar larvae was found to
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 84 | Page
be 0.3344 ± 0.0573 (s = 0.1146).The biomass of the pupae was found to be 0.2934 ± 0.0234 (s = 0.0469). (see
Table.2) The three seasons data were averaged for length, width and calculated biomass of the immature
stages,the results showed no significant diffrences in the average length, average width and calculated average
biomass of the imature stages as [F(2,9) = 0.0184, p = 0.9817], [F(2,9) = 0.2415, p = 0.7903] and [F(2,9) =
0.5254, p = 0.6083] respectively, the averages were plotted in error bars ± standard error, slight intersection
between the error bars meant that average length, average width and calculated average biomass, had little
variations, which might be significantly diffrent seasonaly on Gallus gallus carcass. These results might have
important implications to forensic entomologists, (p > 0.05). And the scatter plot data was made from time and
the averages of the three seasons data were averaged for length, width and calculated biomass of the immature
stages, to see how it varied over time. (See Graph.3)
V. Figures and Tables
Table.1.)
Seasons
Average
tempurature
Average
relative
humidity
(in %)
Average
precipatation
Average
wind
speed
(in ° C)
(in %)
(in
km/hr)
Pre
monsoon
39.4
51.94
0
25.67
Monsoon
33.54
82.47
43.4
15.87
Post
monsoon
30.06
38.31
0
11.5
Fig.1) Morphology of adult Sarcophaga (Parasarcophaga) albiceps male and female
A:Side view adult male genital capsule and 5th sterite: The genitalia of an adult male was dissected and
processed as mentioned in material and methods, viewed under LM. c:cercus;ep:epandrium; prg:pregonite;
j:juxta; v:vesica ; s:surstylus; pht: phallic tube.
B.Female Genitalia:The female genitalia was dissected out and processed as mentioned in materials and
methods. Sternites 6,7and 8,signum,epiproct,cerci and genital tergites 1 and 2 are shown in the figure.ster-6-8:
sternites 6-8; sig:Signum; epi:Epiproct; cer: Cerci; gen-ter-1, 2: Genital tergites 1-2.
C: The 5th sternite: The U- shaped 5th sternite was dessicted out for better visualization,This is also the
conformitory test for identifying the species as shown by LM.
D.Left fore wing of an adult male:The fore wing was visualized under LM and different parts were noted.Sc:
Sub Costal vein; C: Costal vein; R1: anterior branch of radius; R2+3: radial vein; M: media; r-m: radial-medial;
bm-cu: basal-medial-cubital; CuA: anterior cubital.
Fig.2) Morphology of immature stages (3rd instar larvae) of Sarcophaga (Parasarcophaga) albiceps
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 85 | Page
A: Anal division: Posterior view of 3rd instar larvae of Sarcophaga (Parasarcophaga) albiceps, p1-p6: papillae
(1,2,3,4,5,6).
B: Anal division: Posterior view of 3rd instar larvae of Sarcophaga (Parasarcophaga) albiceps, pos: posterior
spiracles (1,2).
C: Anterior of body: Ventral view of post feed 3rd instar larvae of Sarcophaga (Parasarcophaga) albiceps,
t1-t3: thoracic segments (1,2,3).
D: Anterior spiracle.
E: Anterior of body: Anterior end of body of post feed 3rd instar larvae of Sarcophaga (Parasarcophaga)
albiceps, ans1,2: anterior spiracles (1,2).
F: Anal division of body ventral view: Post feed 3rd instar larvae of Sarcophaga (Parasarcophaga) albiceps,
shows iba11 : inter band area 11. There are a total of 12 iba’s in Sarcophaga (Parasarcophaga) albiceps, not
marked.
Fig.3) Life cycle of Sarcophaga (Parasarcophaga) albiceps.
The figure represents the life cycle starting from adult, to different instars, pupae and then imago in a
cyclical way, time taken between the phases are given with average times ± SE on the Gallus gallus carcass for
the three seasons. (Feeding phases on the right, migratory phases on the left and reproductory phase on top).
Fig.4.) Distribution map of Sarcophaga (Parasarcophaga) albiceps in India
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 86 | Page
Sarcophaga (Parasarcophaga) albiceps are marked in the colour red in the physical map of India.
Andhra pradesh, Arunachal Pradesh, Assam, Bihar, Chandigarh, Delhi, Goa, Gujrat, Harayana, Himachal
Pradesh;Kullu;6,000ft, Mizoram, Nagaland, Rajasthan, Sikkim, Tamil Nadu, Tripura, Uttar Pradesh, West
Bengal, Andaman and Nicobar, Karnataka, Kerala, Madhya Pradesh, Manipur, Maharashtra, Orissa, Panjab,
Daman Diu, Pondicherry are marked in light red in the physical map of India.[3]
Graph.1.) The graph shows the time ± SE, for developmental period of Sarcophaga (Parasarcophaga) albiceps
for completion of life cycle in the diffrent seasons and overall annual average on Gallus gallus carcass, West
Bengal, India.
Graph.2.) Graph showing the time ± SE, taken by various stages during development period of Sarcophaga
(Parasarcophaga) albiceps for completion of life cycle in the diffrent seasons and overall annual average on
Gallus gallus carcass, West Bengal, India.
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 87 | Page
Table.2.) Shows the average of time, length, width and calculated biomass ± their respective SE
Stages
Avg.
Hours
(Mean
± SE)
Avg. Length
(in mm)
(Mean ± SE)
Avg. Width
(in mm)
(Mean ± SE)
Avg.Calculated Biomass
(in mm³)
(Mean ± SE)
PRM
M
PSM
PRM
M
PSM
PRM
M
PSM
1st instar
larvae
28.28
±
0.3914
0.3817
±
0.0085
0.3517
±
0.0028
0.3720
±
0.0043
0.0775
±
0.0057
0.0475
±
0.0237
0.0116
±
0.0058
0.0017
±
0.0002
0.0005
±
0.0000
0.0017
±
0.0002
2nd instar
larvae
32.32
±
0.5435
1.0120
±
0.1121
0.9130
±
0.0988
1.0132
±
0.0706
0.2455
±
0.0318
0.1950
±
0.0975
0.2312
±
0.1156
0.0538
±
0.0154
0.0336
±
0.0152
0.0478
±
0.0149
3rd instar
larvae
47.32
±
0.5382
1.8845
±
0.1513
1.6875
±
0.0447
1.7450
±
0.1245
0.5017
±
0.0360
0.3060
±
0.0455
0.4892
±
0.0358
0.3882
±
0.0776
0.1353
±
0.0424
0.3344
±
0.0573
Pupae
114.50
±
1.8058
1.3751
±
0.0483
1.3750
±
0.0368
1.3650
±
0.0500
0.5242
±
0.0145
0.4115
±
0.0564
0.5217
±
0.0140
0.2985
±
0.0237
0.1621
±
0.0483
0.2934
±
0.0234
Graph.3) Show developmental trajectory on the right side as average biometric measurment data’s are plotted
against average time for three seasons ± SE.On the left are the graphs from ANOVA (1-way) data’s averages of
lengths, width and biomass.
VI. Conclusion
The systematic analysis of dipteran colonies on carcass can be a valuable forensic tool in the
determination of PMI (Keh,1985).Many variables that generally influence the rate of development for insects
are not usually incorporated into the insect development model, for estimating PMI. This is largely due to the
lack of base line data of relevant to the non human model implied (Villet.et.al., 2010). However in the current
scheme of models, which uses aging of blow fly larvae, are produced by extrapolation from analysis of the
development rates of larvae,generally reared on single medium: liver (Levot et. Al., 1979; Byrd and Butler,
1996) or ground beef (Anderson,2000; Grassberger et.al.,2003) for human model, so for avian model Gallus
gallus was utilized for the pourpose of rearing and cultivating referance data for the highly thermophilic variety
that is prevalent in India and its extrapolated data might be utilized for human model also.
Since the average length, width and calculated biomass, did not show any significant diffrence,
therefore this current work may be used as reference data for S.albiceps as a forensic indicator, for future studies
on length and width based PMI estimation. In this study, biomass of immature stages was calculated for the
same pourpose as length and width. More study are need to be undertaken.Perhaps in the future a similar study
on diffrent substrates might be conducted, to assess significant variation on diffrent substrate on a spacio-
temporal basis.
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 88 | Page
In conclusion, larvae of Sarcophaga (Parasarcophaga) albiceps seemed to target the head region as the
site of entry in to the carcass.The present results suggest that Sarcophaga (Parasarcophaga) albiceps show no
significant variances in length and width, therefore this can be utilized as a reference data for PMI estimations.
The present result also suggests some limitation in the current scheme of forensic application of data that derive
from a type of animal tissue, other than that on which the larvae has been feeding on. The other being that
mostly till date blow fly larval length and width have been utilized for PMI estimations, current paper also
includes pupa and the forensic indicator being flesh fly instead of blow fly, for generation of base line data. Also
another parameter is added to the arsenal of forensic entomologist, the volume of larvae and pupae are taken
into consideration as its a product of length and width, wich might be useful in PMI estimation and other allied
discplines.
Acknowledgements
The experimental study was conducted in the existing facility and premises of Zoological Survey of
India, Kolkata and the facilities provided by Director, ZSI is kindly acknowledged. Dr. Dhriti Banerjee, is the
head of the Diptera Section at the Zoological Survey of India (ZSI), Kolkata, Dr. Shyamasree Ghosh, is the
Scientific Officer, SBS, NISER, Bhubaneswar, Dr. Waliza Ansar, is the Lecturer in the Dept. of Zoology,
Asutosh College, Kolkata and Mr. Abesh Chakraborty, is the research scholar in ZSI, Kolkata. The authors
express their gratitude to the institutes, ZSI, Kolkata, NISER, Bhubaneswar and Asutosh College Kolkata for
their support.
References
[1]. J.H.Byrd and J.L.Castner (editors), Forensic Entomology: The utility of Arthopods in Legal Investigations:2eds (CRC press,FL:
Boca Raton,2009).
[2]. B.C.Nandi, The Fauna of India and Adjacent countries: Diptera::Sarcophagidae: vol-X (Z.S.I.press, Kol:New Alipore,2002).
[3]. D.D.Mercedes, D.E.Hardy, A catalog of the Diptera of the Oriental region, Suborder Brachycera through Division Aschiza,
Suborder Cyclorrahapha Vol-II,(The University Press of Hawaii,Honolulu,1977).
[4]. R.Senior-White, D.Aubertin, J.Smar, The fauna of British India, including the remainder of theOriental region, Diptera. Family
Calliphoridae.: Vol. VI. (Taylor and Francis, London,1940).
[5]. Flesh flies (Diptera:Sarcophagidae).2014.Electronic reference. Retrived from http://sarcophagidae.myspecies.info/.
[6]. Y.Roskov,T.Kunze, L.Paglinawan, T.Orrell, D.Nicolson, A.Culham, N.Bailly,P.Kirk,T.Bourgoin,G.Baillargeon, F.Hernandez,
A.De Wever, eds (2013). Species 2000 & ITIS Catalogue of Life, 15th August 2015. Digital resource at
www.catalogueoflife.org/col/. Species 2000: Reading, UK.
[7]. T.Pape,& F.C.Thompson,(editors). [1.5, 2013 June]. Systema Dipterorum, Version [1.5]. http://www.diptera.org/, accessed on [15th
August 2015].
[8]. R.Sharma, B.Mitra; 2013. Check-list of Indian flesh flies (Insecta: Diptera: Sarcophagidae)., Electronic reference.
zsi.gov.in/checklist/Indian_sarcophagidae pdf acessed on 2 July 2014.
[9]. KA Meiklejohn, M Dowton, T Pape, JF Wallman, A revised key to the Australian Sarcophagidae (Diptera) with special emphasis
on Sarcophaga (sensulato). Zootaxa, 3680(1), 2013, 148-189.
[10]. H.Bänziger, and T.Pape, Flowers, faeces and cadavers: natural feeding and laying habits of flesh flies in Thailand (Diptera:
Sarcophagidae, Sarcophaga spp.). Journal of Natural History, 38, 2004,1677-1694.
[11]. C.B.Castro, M.D.Garcia, M.I.Arnaldos, and D. González-Mora, Sarcophagidae (Diptera) attracted to piglet carcasses including new
records for Portuguese fauna. Graellsia 66, 2010, 285-294.
[12]. H.Ishijima, Revision of the third stage larvae of synanthropic flies of Japan (Diptera: Anthomyiidae, Muscidae, Calliphoridae and
Sarcophagidae). Japanese Journal of Sanitary Zoology, 18, 1967,47-200.
[13]. Meigen, J.W., Systematische Beschreibung der bekannten europäischen zweiflügeligen Insekten. Fünfter Theil. Schulz, Hamm. xii
+ 412 pp., 1826, 42-54.
[14]. B.Singh and M.Bharti, Some notes on the nocturnal larviposition by two species of Sarcophaga (Diptera: Sarcophagidae). Forensic
Science International, 177,2008, 19-20.
[15]. T.M.Shazia, A.Suhail, and M.J.Yousuf, Systematics and populations of sarcophagid flies in Faisalabad (Pakistan). International
Journal of Agriculture & Biology, 8(6),2006, 809-811.
[16]. P.Fisher, R. Wall, and J.R. Ashworth, Attraction of the sheep blow fly, Lucilia sericata (Diptera: Calliphoridae) to carrion bait in the
field. Bullitein of Entomological Research, 88,1998, 611–616.
[17]. Z.J.O. Adams and M.J.R.Hall, Methods used for the killing and preservation of blowfly larvae and their effect on post mortem
larval length. Forensic Science International, 13,2003,50-61.
[18]. Z.A. El-Moaty and A.E.M.Kheriallah, Developmental variation of the blow fly Lucilia sericata (Meigen,1826) (Diptera:
Calliphoridae) by diffrent substrate tissue types.Journal of Asia-Pacific Entomology,16,2013,297-300.
[19]. G.S.Anderson, Minimum and maximum development rates of some forensically important Calliphoridae (Diptera). Journal of
Forensic Science,45,2000,824-832.
[20]. J.H.Byrd, and J.F.Butler, Effects of temperature on Cochliomyia macellaria (Diptera: Calliphoridae) development. Journal of
Medical Entomology. 33, 1996, 901–905.
[21]. K.Clark, L.Evans, and R. Wall, Growth rates of the blowfly, Lucilia sericata, on different body tissues. Forensic Science
International 156, 2006,145–149.
[22]. D.A.Estrada, M.D.Grella, P.J.Thyssen, A.X.Linhares, 2009. Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae)
developmental rate on artificial diet with animal tissues for forensic purpose. Neotropical Entomology, 38, 2009, 203–207.
[23]. M.Grassberger, E. Friedrich, and C. Reiter, The blow fly Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae) as a new
indicator in Central Europe.International Journal of Legal Medicine. 117, 2003,75–81.
[24]. B. Greenberg, and J.C. Kunich, Entomology and the Law: Flies as Forensic Indicators. (Cambridge University Press, Cambridge
,2002).
[25]. M . Hall and R . Wall,. Myiasis of humans and domestic animals. Advanced Parasitology. 35, 1995,258–334.
Developmental analysis of immature stages of Sarcophaga (Parasarcophaga) albiceps Meigen,1826 ..
DOI: 10.9790/2380-08827989 www.iosrjournals.org 89 | Page
[26]. C.C.Hwang, Urban Ecology of Necrophagous Flies in Greater London. (Ph. D.Thesis) King's College, London, 2004.
[27]. G.W.Levot, K.R.Brown, and E. Shipp, Larval growth of some calliphorid and sarcophagid Diptera. Bullitein of Entomological
Research 69, 1979,469–475.
[28]. K.G.V.Smith,. A Manual of Forensic Entomology. British Museum (Natural History). (London, and Cornell University Press,
Ithaca, New York. 1986).
[29]. M.J.Norušis,. SPSS 13.0 Guide to Data Analysis. (Prentice Hall, Inc., New Jersey, USA, 2005).
[30]. P.Nuorteva, Sarcosaprophagous insects as forensic indicators. In: C.G.Tedeschi, W.G. Eckert, L.G. Tedeschi,. (Eds.), Forensic
Medicine: A Study in Trauma and Environmental Hazards, vol. 2. (Saunders, Philadelphia,. 1072–1095,1977).
