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85
ISSN 2320-4311
PROMMALIA. Vol. III, 2015
Chakraborty, A., Ghosh, S., Ansar, W. and Banerjee, D. 2015. Thermophilic Pupal Endoparasitoids:
Brachymeria minuta (Hymenoptera: Chalicididae) on forensic indicator Sarcophaga
(Parasarcophaga) albiceps. PROMMALIA, III, 85-94.
http://zoobank.org/urn:lsid:zoobank.org:pub:1F1BA525-2887-44EE-8A2F-4D9461664F6E
Date of Publication: 5th October, 2015
THERMOPHILIC PUPAL ENDOPARASITOIDS: BRACHYMERIA MINUTA
(HYMENOPTERA: CHALICIDIDAE) ON FORENSIC INDICATOR SARCOPHAGA
(PARASARCOPHAGA) ALBICEPS
ABESH CHAKRABORTY, SHYAMASREE GHOSH*, WALIZA ANSAR**
AND DHRITI BANERJEE
Email: abeshc1@gmail.com
Zoological Survey of India, M-Block, New Alipore, Kolkata-700 053.
*School of Biological Sciences, National Institute of Science, Education and Research (NISER),
Bhubaneswar- 751005, India.
**Dept of Zoology, Post Graduate Dept, Asutosh College, 92, S.P. Mookerjee Road Kol-700026.
ABSTRACT
The blow flies and the flesh flies are the most widely used model organisms utilized in the immature
insect development model. Since, their immature stages are used to estimate the post mortem interval
(PMI). Therefore, this study investigates the effect of parasitoid attack on pupal development of the
flesh Sarcophaga (Parasarcophaga) albiceps. Host specificity = 65.62 % and avg. yearly
parasitoidism = 50.93 %.The result might be utilized by forensic entomologist to properly decipher
PMI estimates from reference data, form the thermophilic variety of forensic indicators prevalant in
India
KEYWORDS
Forensic Dipterology, Thermophilic, Pupal, endoparasitoid, Sarcophaga albiceps, India.
INTRODUCTION
The objective of this study was to report the occurence of pupal endoparasitoidism of Brachymeria
minuta (Linnaeus, 1767) (Hymenoptera: Chalicididae) on pupa of Sarcophaga (Parasarcophaga)
albiceps Meigen, 1826 (Diptera: Sarcophagidae), a fly of medical, veterinary and forensic importance.
In India, meterologically a year is divided into 3 seasons (pre monsoon, monsoon and post monsoon).
Chicken meat was used in the all the season as bait to attract insects and malise trap was used for their
capture. In the study 32 pupae of S. (P) albiceps were obtained over a period of 1 year (including 3
seasons), out of which 21 yielded the parasitoid Brachymeria minuta (Linnaeus, 1767). Thereby
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showing average host specificity of 65.62 % and average yearly level of pupal parasitoidism of
50.93%.
Since Sarcophaga (Parasarcophaga) albiceps Meigen, 1826 (Diptera: Sarcophagidae), a fly of
medical veterinary and forensic importance, and has also been reported from dead animals (Senior-
White, 1940) previously from earlier reports. The current experimentation was made on chicken meat
(as bird model). B. minuta shows host specificity and also a specific rate of development, and all these
factors may help forensic dipterology in assessing disruptions of life cycle and also S. albiceps have
been known to infest dried fish and fish products, due to their need for larvipositing in places of fish
drying, they may be potential mechanical carrier of disease due to their association with filth and
cause myasis in humans and animals, therefore this data may also be useful in bio control of these
nuisances.
The parasitoid infestation of dipterans in the various stages of their life cycle seems to be species
specific, as well as specific to stages of life cycle. The parasitoid infestation of forensic dipterans.,viz.
Lucilia sp. (Diptera: Calliphoridae) and Sarcophaga sp. (Diptera: Sarcophagidae) by Alysia
ridibunda, (1829) (Hymenoptera: Barconidae) on beef substrate was reported by (Lindquist, 1940)
and Olivia, 2008. Effects of temperature on parasitoids in Brazil were reported by (Almeida et al,
2002). This emerging trend of micro-ecology, have found potential use in forensic dipterology. The
corpse is treated as substrate for growth of diffrent forensic dipterans and in this case S. (P) albiceps
grown on chicken meat for 3 seasons (pre monsoon, monsoon and post monsoon). As corpse
biochemistry varies across different classes of vertebrates, this parasitoid attack occurred on S. (P)
albiceps pupa, growing on chicken meat (bird model).
The study was conducted at the ZSI, Kolkata campus, and was conducted for 3 seasons (pre monsoon,
monsoon and post monsoon). The climatic regimes in West Bengal (India) are divided into 3 seasons
of each four months, which are pre monsoon (March-June), monsoon (July-October) and post
monsoon (November-February). Last months of each seasons in 2013-2014 were taken for conducting
the experiment. Malise trap was utilized for capture of the dipteran specimens and some parasitoids
(adults). Suitable pupation material was provided for, in this case a cloth was used to help the larvae
pupate, and then the contents of that piece of cloth, after a few days B. minuta was observed
ovipositing on the pupa, now the contents of the cloth were taken along with the un-infested and
infested pupa into a glass container.
The trap was checked regularly and notes on the appearance time of dipterans, their ovipositing or
larvipositing time, larval developmental time, pupal developmental time to arrival of hymenopteran
parasitoides and emergence of parasitoides from host pupa and photographs taken by the Nikkon P
520 camera. The adult Sarcophagidae and Chalcididae specimens were collected from malise traps
and killed by ethyl ether, and preserved in 70% ethyl alcohol vials of 5 ml. The larvae, pupae, and
infested pupae were kept in 70% alcohol, after diping them in 10% warm KOH sol. for 3 mins.
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MATERIALS AND METHODS
STUDY SITE
The study was conducted in Kolkata, ZSI, Latitude: 22° 30' 51.6888" and Longitude: 88° 19' 30.5256"
recorded by GPS meter. The material for this work was from a dead chicken carcass to attract flies.
COLLECTION
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 trap was used to
capture the flies in one part of ZSI, Kolkata, in 2013-2014. The parasitoides were obtained differently,
as the post feed maggots (larvae of 3rd instar) starts their migration away from the carcass into suitable
pupation material and in this case a cloth was used to help the larvae pupate, and then the contents of
that piece of cloth, after a few days B. minuta was observed ovipositing on the pupa. The contents of
the cloth were taken along with the pupa and infested pupa into a glass container, to see progression
of normal development.
SAMPLING
The trap was checked regularly and notes on the appearance time of dipterans, their ovipositing or
larvipositing time, larval developmental time, pupal developmental to arrival of hymenopteran
parasitoides and emergence of parasitoides from S.(P) albiceps pupa and photographs taken by the
Nikkon P-520 camera.
PRESERVATION
The adult Sarcophagidae and Chalcididae specimens were collected from malise traps and killed by
ethyl ether, and preserved in 70% ethyl alcohol vials of 5 ml. The larvae, pupae, and infested pupae
were kept in 70% alcohol, after diping them in 10% warm KOH sol. for 3 mins.
IDENTIFICATION AND DESCRIPTION
Morphogenitic keys of certified reference materials for identification of specimens were used for the
identification of host and its parasitoid and the method adopted by Nandi. B. C. (2002) and for
parasitoid identification the specimens were sent to S. Sheela (Hymenoptera Section, Z.S.I., Kolkata).
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ANALYTICS
The prevalance of parasitoidism was calculated by the method pioneered by Marchiori, 2002, with
minor modifications for the calculation of host specificity. The prevalance of parasitoidism was
calculated by the following formula:
Level of Parasitoidism = (Emergence of Parasite from pupae /
Total Emergence of normal host from pupae) x 100.
The specificity to the host was calculated by the following formula
Host specificity = (Emergence of B. minuta from Pupa of S. (P) albiceps /
Total pupa produced by S. (P) albiceps) x 100.
The ecological data was processed from the metrological data observed at the site and averaged to
give seasonal development data till pupation of Sarcophagidae larvae to emergence of adult
Brachymeria minuta (Linnaeus, 1767), for the three seasons observed. Light microscopy via Leica M-
205 Stereozoom dissecting microscope, where used the specimens were photo documented, dissected
etc.
RESULTS
The study was conducted in climatic regimes in India (West Bengal) divided into 3 seasons of each
four months, which are Pre monsoon (March-June), Monsoon (July-October) and Post monsoon
(November-February). Last two months of each season were taken for conducting the experiment.
PREMONSOON (MARCH TO JUNE)
After 7 ± 1 days the Sarcophagidae larvae went in to pupation, at ambient air temperature of 38 ± 3
(in °C), Avg. precipitation (in %)- 0 and Avg. relative humidity (in %) ~ 52 ± 7 and Avg. wind speed
(in Km) ~ 24 ± 7. Brachymeria minuta (Linnaeus, 1767) showed host specificity over the species of
Sarcophaga (Parasarcophaga) albiceps Meigen, 1826 (Diptera: Sarcophagidae), but other dipteran
fauna on the chicken cadaver were not touched by this Hymenopterans. The level of parasitoidism
was an astounding 90%., viz. 18 out of the 20 pupas produced Brachymeria minuta (Linnaeus, 1767),
only 2 pupas produced Sarcophaga (Parasarcophaga) albiceps Meigen.
MONSOON (JULY TO OCTOBER)
After 7 ± 1 days the Sarcophagidae larvae went in to pupation, at ambient air temperature of 33.5 ±
3.5 (in °C), Avg. precipitation (in %) ~ 46.5 ± 11.5 and Avg. relative humidity (in %)- 82.5 ± 7.5 and
Avg. wind speed (in Km) ~ 16 ± 4. Brachymeria minuta (Linnaeus, 1767) showed host specificity
over other species of Sarcophaga (Diptera: Sarcophagidae), flies and other dipteran fauna on the
chicken cadaver were also not touched by this Hymenopterans. The level of parasitoidism was 20 %.,
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viz. 5 out of the 1 pupas produced Brachymeria minuta (Linnaeus, 1767), only 2 pupas produced
Sarcophaga (Parasarcophaga) albiceps Meigen, 1826 and other 2 didn’t develop.
POSTMONSOON (NOVEMBER TO FEBRUARY)
After 7 ± 1 days the Sarcophagidae larvae went in to pupation, at ambient air temperature of 30 ± 5
(in °C), Avg. precipitation (in %) ~ 0 and Avg. relative humidity (in %) ~ 38.5 ± 3.5 and Avg. wind
speed (in Km) ~ 11.5 ± 3.5. Brachymeria minuta (Linnaeus, 1767) showed the same host specificity
over the species of Sarcophaga (Diptera: Sarcophagidae), flies and other dipteran fauna on the
chicken cadaver were not touched by this Hymenopterans. The level of parasitoidism was 42.8 %.,
viz. 3 out of the 7 pupas produced Brachymeria minuta (Linnaeus, 1767), only 2 pupas produced
Sarcophaga (Parasarcophaga) albiceps Meigen,1826 and other 2 didn’t develop. [Graph-1]
Seasonal variation of Parasitism level of Brachymeria
minuta on pupae of Sarcophaga albiceps
0
10
20
30
40
50
60
70
80
90
Premonsoon Monsoon Postmonsoon
Level of Parasitism (in %)
Seasons
Seasonal variation of Parasitism level of Brachymeria m inuta
on pupae of Sarcophaga al biceps
Graph-1
Brachymeria minuta (Linnaeus, 1767) (Hymenoptera: Chalicididae) from pupa of Sarcophaga
(Parasarcophaga) albiceps Meigen, 1826 (Diptera: Sarcophagidae, takes around 20 ± 2 days, 2 hrs.
(479.6 ± 48.2 hrs.) to complete its life cycle. We observed that around 252 ± 4 hours from
larviposition to pupation of S. albiceps, parasitoid attack came exactly around 252 ± 4 hours or
around 10.5 days, when Brachymeria minuta (Linnaeus, 1767) (Hymenoptera: Chalicididae) inserted
its own egg to develop into an adult. The entire life cycle of Sarcophaga (Parasarcophaga) albiceps
on chicken generally completes around 324 ± 12 hrs. But due to parsitoid attack, additional 155.6
hours or around 6 days, 48 hrs was added to the count, thus extending the normal PMI calculations to
20 ± 2 days, 2 hrs (479.6 ± 48.2).(Fig-2)
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Fig.2: Shows the Normal life cycle of S. (P) albiceps and pupal parasitoid
infestation
Normal life cycle of S. (P) albiceps Parasitoid infested life cycle of
(in Bird carcass as model) S. (P) albiceps (in Bird carcass as model)
According to the seasonal variation of parasitoidism level of Brachymeria minuta (Linnaeus, 1767)
(Hymenoptera: Chalicididae) on pupa of Sarcophaga (Parasarcophaga) albiceps Meigen, 1826
(Diptera: Sarcophagidae) seems to be more active in the summer period (pre monsoon), viz. 90%,
followed by winter (post monsoon), viz.42.8% that’s almost half of summer period and their activity
almost negligent in the rainy period (monsoon), viz. 20% that’s almost half of winter period. Only in
the monsoon and post monsoon the 2 pupa’s each didn’t develop. (Table-1 and 2).
Table-1: Shows ecological parameters along with parasitoid (B. minuta) infestation levels on S.
(P) albiceps pupa for three seasons
Seasons Level of
Parasitoi
dism
(%)
Total
pupa of
S.(P)
albiceps
Emergen
ce of
S.(P)
albiceps
Emergen
ce of
B.
minuta
Pupa
that
didn’t
develop
Average
ambient
temp
(°C)
Average
Precipat
ation
(%)
Average
Relative
Humidit
y
(%)
Average
Wind
speed
(%)
Pre
Monsoon
90 20 2 18 0 38.94 0 52.10 25.94
Monsoon 20 5 2 1 2 33.75 43.10 82.30 15.90
Post
Monsoon
42.8 7 3 2 2 29.54 0 38.13 11.04
91
Table-2: Shows the developmental times of B. minuta on pupa of S. (P) albiceps for three seasons
Stages/Seasons Pre Monsoon
(Duration of
stages in hrs)
Monsoon
(Duration of
stages in hrs)
Post Monsoon
(Duration of
stages in hrs)
Average
yearly
(Duration of
stages in
hrs)
Pupa of S.(P) albiceps 162 198 180 180± 18
Laying of Egg of B. minuta on
Pupa of S.(P) albiceps
54 66 60 60±6
Larvae of B.minuta on Pupa of
S.(P) albiceps
89.8 109.8 100 99.8±10
Pupa of B.minuta on Pupa of
S.(P) albiceps
80.7 98.7 90 89.8±9
Adult of B.minuta on Pupa of
S.(P) albiceps
44.9 54.9 50 49.9±5
Total life cycle of Parasitoid on
on Pupa of S.(P) albiceps
431.4 527.4 480.0 479.6±48.2
Almost in all the pupas which were infested by the parasite a pattern followed, the parasite seemed to
have oviposited on the 2nd and 3rd segments of pupa, perhaps this might be a possible binding site for
Brachymeria minuta (Linnaeus, 1767) (Hymenoptera: Chalicididae) on pupa of Sarcophaga
(Parasarcophaga) albiceps Meigen, 1826 (Diptera: Sarcophagidae).Also the pupas infested by
parasitoid shrunk in size, when compared to pupas that underwent normal development. (Fig. 3)
Fig. 3: A: Shrunken S. (P) albiceps Pupal case infested by Parasitoid and B: Normal Pupal case.
In order to characterize the identified fly and parasitoid, we have taken light microscopic images of
the third instar larva and the male genitalia of S. (P) albiceps (unpublished data). (Fig-2 and 1).
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Fig 1: Shows B. minuta habitus
In the study a total of 32 pupae of S. (P) albiceps were obtained over a period of 1 year (including 3
seasons), out of which 21 yielded the parasitoid Brachymeria minuta (Linnaeus, 1767). Thereby
showing average host specificity of 65.62 % and average yearly level of pupal Parasitoidism of
50.93%.
DISCUSSION
The current study throws light on probable disruption rate of the very thermophilic variety of
Sarcophaga (Parasarcophaga) albiceps Meigen, 1826 (Diptera: Sarcophagidae), when attacked by
parasitoid Brachymeria minuta (Linnaeus, 1767). Current data suggest that parasitoidism vary
seasonally, peaking in summer, temperature seems to be directly proportional to development of
Brachymeria minuta (Linnaeus, 1767) on pupa of Sarcophaga (Parasarcophaga) albiceps Meigen,
1826, whereas humidity seems to be inversely proportional to development Brachymeria minuta
(Linnaeus, 1767) on pupa of Sarcophaga (Parasarcophaga) albiceps Meigen, 1826. As this field is
still virgin, therefore there a tremendous scope for future work in this sphere.
B. minuta shows host specificity and also a specific rate of development, all these factors may help
forensic science in establishing pmi, tsd, ton and also S. albiceps have been known to infest due to
their need for larvipositing in places of fish drying, they may be potential disease transmitting vectors
due to their association with filth and cause myasis in humans and animals, therefore this data may
also be useful in biocontrol of these nuisances. Still more study and observations are needed to
correlate the Level of Parasitoidism (%) with average ambient air temperature, average precipitation,
average relative humidity and average wind speed. But current data suggest that parasitoidism vary
seasonally, peaking in summer, temperature seems to be directly proportional to development of
Brachymeria minuta (Linnaeus, 1767) on pupa of Sarcophaga (Parasarcophaga) albiceps Meigen,
1826, whereas humidity seems to be inversly proportional to development Brachymeria minuta
Brachymeria minuta
(Linnaeus,1767)
(Hym e nop tera: C halicid ida e
Thorax
Antenna
Head
Leg
?Abdomen?Abdomen
93
(Linnaeus, 1767) on pupa of Sarcophaga (Parasarcophaga) albiceps Meigen, 1826. (Table. 2) As this
field is still virgin, therefore there a tremendous scope for future work in this sphere.
B. minuta shows host specificity and also a specific rate of development, all these factors may help
forensic science in establishing post mortem interval, time since death, time of negligence and also S.
albiceps have been known to infest due to their need for larvipositing in places of fish drying, they
may be potential disease transmitting vectors due to their association with filth and cause myasis in
humans and animals, therefore this data may also be useful in bio control of these nuisances.
Conflict of Interest: The authors hereby declare no conflict of interest.
ACKNOWLEDGEMENT
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. S.
Sheela is Scientist- C of the Hymenoptera Section at the Zoological Survey of India (ZSI) is
acknowledged for her kind help in identification of the parasitoid, 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
Amat. E.2012. Notes on Necrophagous flies (Diptera: Calyptratae) associated to fish carrion in
Colombian Amazon. Acta Amaz., 40: 2.
Ferriere.H.C and Kerrich. J. G. 1958. Handbook for identification of British insects. (Hymenoptera:
Chalcidoidea). Vol: VIII. (2a): 11-16.
Marchiori.C. H, Pereira.L. A, Filho. O. M. S, Riberio. L. C. S and Borges.V. R. 2002. First occurence
of parasitoid Spalangia endius (Walker) (Hymenoptera: Pteromalidae) in pupae of Peckia
chrysostoma (Wiedemann) (Diptera: Sarcophagidae) in Brazil, Biotemas, 15 (2): 75-78.
Marchiori. C. H. and Borges. V. R. 2013. First occurrence Tachinobia sp (Hymenoptera: Eulophidae)
as a parasitoid of Sarcodexia lambens Walker (Diptera: Sarcophagidae). International
Journal of Advanced Technology and science: 1-3.
Mercedes D. D., Hardy. D. E. 1977. A catalog of the Diptera of the Oriental region, Suborder
Brachycera through Division Aschiza, Suborder Cyclorrahapha (II), Family Sarcophagidae,
The University Press of Hawaii, Honolulu: 557-583 pp.
Mitra, B., Sharma, R. M. 2013. Check-list of Indian flesh flies (Insecta: Diptera: Sarcophagidae);
www.zsi.gov.in/checklist/Indian_sarcophagidae.pdf available from 2nd July, 2013.
94
Mora.M. A. R, Buenaventura, E, Gomes, P. L. M and Amat, E. 2012. Updated checklist and new
records of Calyptratae carrion flies (Diptera: Schizophora) from Valle de Aburrá and other
localities in Colombia. Entomotropica, 27 (1): 27-35.
Nandi, B. C. 2002. Fauna of India: Diptera: Sarcophagidae. Zoological Survey of India. 10: 270-274
Olivia. A. 2008. Parasitoid wasp (Hymenoptera) from puparia of Sarco saprophagus flies (Diptera:
Calliphoridae and Sacrophagidae) in Buenos Aires, Argentina. Rev. Soc. Entomol. Argent. 67
(3-4): 139-141.
Senior-White, R., Aubertin, D., Smart, J. 1940. The fauna of British India, including the remainder of
the Oriental region, (Diptera: Calliphoridae), 6: 208-281.
