Sarcophaga ruficornis (Diptera: Sarcophagidiae) a …€¦ · · 2017-01-03The Journal of Zoology Studies Vol. 3 No. 6 2016 Journalofzoology.com Page 6 Table 3: Result of Tukey

The Journal of Zoology Studies

Vol. 3 No. 6 2016 Journalofzoology.com

Page 1

The Journal of Zoology Studies 2016; 3(6): 01-08

ISSN 2348-5914

JOZS 2016; 3(6): 01-08

JOZS © 2016

Received: 28-10-2016

Accepted: 06-12-2016

Kamal Adhikari

Post Graduate Student,

Gauhati University,

India.

Bulbuli Khanikor

Assistant Professor,

Gauhati University,

India.

Riju Sarma

Research scholar,

Gauhati University,

India.

Sudarshana Mahanta

Research scholar,

Gauhati University,

India.

Jatin Kalita Professor,

Gauhati University,

India.

Corresponding Author:

Bulbuli Khanikor

Assistant Professor,

Gauhati University,

India.

Study on the life history and protein content of Sarcophaga ruficornis (Diptera: Sarcophagidiae) a

forensically important insect

Authors: Kamal Adhikari, Bulbuli Khanikor, Riju Sarma, Sudarshana Mahanta, Jatin Kalita

Abstract

Since past few decades insects have been serving as an important tool in forensic entomology

i.e. in determining the time elapsed since death. The present investigation aims at studying one

of the primary colonizer of carcass namely Sarcophaga ruficornis (Diptera: Sarcophagidiae) in

Guwahati, Assam. The growth and development of S. ruficornis like other insects depends

strictly on the climatic conditions prevailing in the area and level of exposure of the corpse. For

laboratory culture of the flesh fly S. ruficornis, chick liver was taken as bait. The total protein

content of different developmental stages of S. ruficornis and the fresh as well as rotten chick

liver was determined by using the method of Lowry et al. The developmental time of S.

ruficornis was found as 25±3 days during the investigation period (23±1 days during May and

28±2 days during the month of February). The protein content of the liver was found to decrease

during its decomposition stage and the protein content of the developing stages of the fly was

found to increase linearly.

Keywords: Carcass, Sarcophaga ruficornis, Protein, Diptera: Sarcophagidiae, Insect

1. Introduction

Arthropods are among the most evolved groups of animals on earth. So they are found almost

everywhere on earth. Insects like other arthropods play a crucial role in different fields of

modern science like forensic entomology. Knowledge of the distribution, biology and behavior

of insects found at a crime scene can provide information on when, where and how the crime

was committed (Kashyap and Pillai, 1989 [1]

; Anderson and Carvenka, 2001 [2]

; Hall, 2008 [3]

).

Concepts of algor mortis, rigor mortis, and livor mortis play an important role during the first

few hours of death and hardly can be determined up to 3 days by these methods. However, all of

these parameters are affected by many other factors such as body size, age, illness, exertion

period to death etc. and become less valuable as time passes (Simpson and Knight 1985 [4]

;

Henssge et al, 1995 [5]

). Insects are never affected by all these parameters so they play a major

role in determining the post mortem interval, among which flies are of primary significance.

These insects feed on the corpse, oviposit and the larvae hatches into successive instars and

finally emerges as an adult. Blow flies and flesh flies are among the first colonizer (Luna et al.,

2001 [6]

; Bharti and Singh, 2003 [7]

). The study of their first mature maggots can provide the data

of time elapsed since death. Attraction of the arthropod species varies according to the

decomposition state of the corpse (such as fresh, bloat, decay, putrefaction, mummification, and

skelotization). A particular species never stay in the corpse during the whole process of

decomposition (Bornemissza, 1957 [8]

; Braack, 1981 [9]

). There is a succession of species of

arthropods. Each species stay only for a limited period of time (Anderson, 2009 [10]

).



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In the present study a forensically important species,

Sarcophaga ruficornis (Diptera: Sarcophagidae) was

chosen to study the life cycle by keeping them in a

close culture chamber and provided chick liver as bait

within the chamber.

Generally S. ruficornis is found abundant in carcass in

the early stage of decomposition. It is generally not

found after 3-4 days. It is a medium sized to large sized

fly. Front broad in the female somewhat narrower in

the male. Distal portion of the arista is bare. Abdomen

consists of 4 visible segments. External genitalia in

male are prominent. The puparium is reddish brown

and ovoid in shape. Mode of reproduction of

Sarcophaga is ovoviviparous, i.e. they lay first instar

maggot on the flesh.(Sukontason et al.,2014[11]

).

The purpose of studying the life cycle of a single

species was to get a more accurate and a firm report of

the life cycle. All the life cycle of the necrophagous

flies is more or less similar with the life cycle of

Sarcophaga ruficornis.

2. Methodology

2.1. Obtaining the specimen Sarcophaga ruficornis was collected by exposing a bait

of broiler. Flies were attracted to the bait. They

oviposit in the bait and the maggots grow into

successive instars and finally reached the pupal stage.

Pupae at this stage were collected in an insect proof

container and waited till they hatch.. After few days of

adult fly emergence, few other species of the family

Muscidae, Calliphoridae and Sarcophagidae that also

emerged simultaneously in the cage such as were

removed. Thus, a pure culture of Sarcophaga

ruficornis was achieved and maintained solely by

providing chick liver as bait. The species was

identified as Sarcophaga ruficornis by the experts from

Zoological Survey of India, Kolkata. In the present

study it was found that the species of Sarcophaga were

more abundant in the late winter than the species of

other forensically important insects. So their collection

was easier.

Fig 1: Broiler exposed for decomposition showing infestation and larviposition by flies.

2.2. Culture of S. ruficornis:

The first generation of the adults that emerged was

collected by allowing a bait for larviposition as

mentioned earlier. These adults were again provided

with the chick liver as bait to continue their life cycle.

The culture chamber was made of a glass container of

which the upper open portion was covered with a

mosquito net. Sufficient dry mud with sand was

provided at the bottom of the chamber so that the pre

pupa does not move to and fro in search of the suitable

place for pupation.

Fig 2: Culture chamber



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Fig 3: 2

nd instar maggot of Sarcophagaruficornis

growing in culture.

In this way, four such generations were maintained.

During the culture period no other insects were allowed

to mix with the culture.

During the experimental period broiler was also

exposed separately in the open condition and its

decomposition along with the infestation by different

flies were observed. The broiler was kept inside a wire

gauze of considerable perforation so as to prevent the

attack of other predators in open condition.

Here it was found that the fly life cycle was shorter in

wild than in culture in the laboratory. Due to its small

size the broiler was found to skeletonize after 8 days of

exposure period. After skeletonization, normal visitors

of the corpse disappeared.

2.3 Estimation of Protein

Estimation of protein content was done following the

method of Lowry et al., (1951) [12]

. The total protein of

1st instar larvae, 3

rd instar larvae, pupae and adults of

the fly were taken as the sample. Protein of fresh and

decomposing liver on 5th

day of decomposition were

also estimated following the same method.

2.4. Statistical Analysis

The Tukey test of the protein content of the meat

sample and different instars of S. ruficornis were done

with the help of SPSS (Version 16) software.

3. Results and Discussion

Fig 4: Life cycle of Sarcophaga ruficornis



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Table 1: Duration of different developmental stages of Sarcophaga ruficornis in different temperature.

S. no Time Average Developmental stages Total

Temperature 1st instar 2nd 3rd Pre- Pupa Adult lengh of

(0C) (Days) instar instar pupal (Days) (Days) Life

(Days) (Days) stage cycle(Da (Days) ys)

1 February 28±1 3±1 2±1 2±2 3±2 13±1 5±1 28±2

2 March 30±1.33 2±1 2±1 2±1 3±2 12±2 5±2 26±2

3 April 31±2 2±1 2±1 2±1 2±1 11±1 5±1 24±1

4 May 32±2 2±1 2±1 2±1 1±1 11±1 5±1 23±1

As shown in the table (Table1) above the

developmental period of the fly was found to strictly

dependent on the temperature and level of exposure of

the corpse. In the month of April and May when the

temperature was higher the developmental of the fly

was found to accelerate. Whereas in the month of

February when the temperature was lower the flies

took longer time to develop. This result was in

conformity with the findings of earlier researchers

(Byrd and Butler, 1997 [13]

; Wells and Kurahashi, 1994 [14]

; Boatright et al,. 2010 [15]

).

Flies oviposit only when the carcass is fresh (Archer,

2003 [16],[17]

; Hall et al., 1993 [18]

). This has been found

true in the present study. But in the favorable season, in

case of the small carcass the fly infestation was

recorded maximum and therefore the oviposition rate

was also found maximum. The huge number of the

larvae was found to feed on the carcass voraciously

and skeletonize it within a very short period of time.

Moreover, the oviposition time was observed upto

early bloated period. Flies were observed to visit the

carcass till later part of the early bloated stage but they

were not found to deposit maggot on it. The flies were

rarely seen in the late decay phase and almost never

seen on the dry phase.

During the investigation period, the culture of the flesh

fly Sarcophaga ruficornis was almost successfully

completed. The adult fly lived for 3-7 days during

which it deposit maggot in the bait until it was fresh.

After 3-4 days the flies were resting on the wall of the

culture chamber and avoided the bait. Life cycle

depended on the temperature and humidity. It was

observed that the life cycle which was studied in

February was longer than the life cycle that was

studied in May. Likewise with the advent of summer

life cycle shortened. The average life cycle in culture

was found to be 25±3.

It was also found that the fly grown in culture had a

longer life cycle than the fly growing in the wild.

3.1 Protein Estimation Proteins of different stages of S. ruficornis were

determined by following the method described by

Lowry et al., (1951) [11]

.

Table 2: Showing the protein content of developmental stages

Sl No. Sample Protein(mg/ml tissue ±SE)

1 Fresh Liver 9.8±0.08

2 Rotten Liver 8.4±0.01

3 1st instar 7.0±0.38

4 Last instar 11.84±0.05

5 Pupa 20.3±0.25

6 Adult 17.22±0.08



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Fig 5: Bar diagram showing the relationship between standard protein and optical density

Fig 6: Bar diagram showing the protein content of different developmental stages

Fig 7: Bar diagram showing the gradual decrease of protein content during decomposition and the increase of the

protein content in the developing stages of Sarcophaga ruficorins



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Table 3: Result of Tukey test for the protein content

Serial No I group J group Mean difference(I-

J) Significance

95% confidence intervel

Lower

bound

Upper

bound

1 Fresh liver

rotten liver 1.39333* .000 1.2587 1.5280

1st instar 2.73667* .000 2.6020 2.8713

Last instar -2.02333* .000 -2.1580 -1.8887

pupa -10.50333* .000 -10.6380 -10.3687

adult -7.41000* .000 -7.5447 -7.2753

2 Rotten liver

fresh liver -1.39333* .000 -1.5280 -1.2587

1st instar 1.34333* .000 1.2087 1.4780

Last instar -3.41667* .000 -3.5513 -3.2820

pupa -11.89667* .000 -12.0313 -11.7620

adult -8.80333* .000 -8.9380 -8.6687

3 1st instar

fresh liver -2.73667* .000 -2.8713 -2.6020

rotten liver -1.34333* .000 -1.4780 -1.2087

Last instar -4.76000* .000 -4.8947 -4.6253

pupa -13.24000* .000 -13.3747 -13.1053

adult -10.14667* .000 -10.2813 -10.0120

4 Last instar

fresh liver 2.02333* .000 1.8887 2.1580

rotten liver 3.41667* .000 3.2820 3.5513

1st instar 4.76000* .000 4.6253 4.8947

pupa -8.48000* .000 -8.6147 -8.3453

adult -5.38667* .000 -5.5213 -5.2520

5 Pupa

fresh liver 10.50333* .000 10.3687 10.6380

rotten liver 11.89667* .000 11.7620 12.0313

1st instar 13.24000* .000 13.1053 13.3747

Last instar 8.48000* .000 8.3453 8.6147

adult 3.09333* .000 2.9587 3.2280

6 Adult

fresh liver 7.41000* .000 7.2753 7.5447

rotten liver 8.80333* .000 8.6687 8.9380

1st instar 10.14667* .000 10.0120 10.2813

Last instar 5.38667* .000 5.2520 5.5213

pupa -3.09333* .000 -3.2280 -2.9587

* The mean difference is significant at the 0.05 level.

From result of Tukey test (table 3) for the protein

content of fresh and rotten liver along with the

different developmental stages of S. ruficornis, the

values were found significantly different from each

other.

The amount of protein content in these flies was found

to totally dependent on the protein content of the

carcasses. We know that liver contains relatively more

proteins than other macromolecules (Guinez et al.,

2011 [19]

)



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The protein content of insects depends on the

metamorphosis stage; adults usually have higher

protein content than other instars (Ademolu et. al.

2007) [20]

.

But in the present study the result did not match

exactly with the findings of Ademolu et al. (2007) [20]

From the experiment it was found that the pupal stage

contained the highest amount of protein among all the

developmental stages. Sarcophaga ruficornis has an

ovoviviparous mode of reproduction, in which eggs

hatch into first instar maggot in the female

reproductive system and the 1st instar maggots are

deposited in the carcass. Generally S. ruficornis adults

feed on the sap of the flesh (as the fly is in the culture

chamber). The carcass in the summer dries up very

fast. So, possibly it may happen that the carcass was

not in the state in which the flesh fly feed (Byrd and

Castner, 2001)[21].

First instar maggot after being laid

on carcass feed voraciously and attained successive

instars. During this feeding period they store sufficient

nutrients for the adult stage. From the experiment it

was found that the total protein contents accumulate in

the larvae gradually and reach its peak during the pupal

stage (Fig.6 and Table.2).

When a graph was plotted to show the total protein

content of different developmental stages of S.

ruficornis it was found that the total protein content

rose linearly till the pupal stage and again fell to some

extent in the adult stage. From this finding, it can be

inferred that the quantity of protein that was lost in

adult was utilized during the pupal to adult

transformation process.

An attempt was also made to correlate the protein

content in the fresh liver where the first instar maggots

were usually thrive and rotten liver where the last

instar maggots were usually found. From the

experiment it was found that protein content of the

liver degraded gradually during decomposition process

and the protein content of the developing stages of the

flies increased gradually during development period.

The total protein content of pupa was found to be

20.3±0.25, which was relatively higher in comparison

to the other developmental stages (Fig.6 & Fig.7).

From these findings it can be inferred that the protein

content of different developmental stages of the flies

was dependent on the protein profile of the carcass.

4. Conclusion

Forensic entomology is evolving as an inevitable

branch of forensic studies. The insects that colonize the

carcass serve as clock for the estimation of post

mortem interval. Here in the investigation life cycle of

a forensically important insect Sarcophaga ruficornis

was studied. To quantify the amount of protein present

in the carcass and the growing larva, protein of various

developmental stages and the larvae were estimated

using the method of Lowry et al., (1951). The results

of the investigation reveal that the time required for the

development of a fly varies within a narrow range of

25±3 during the study period. Protein content of the

developing stage increased linearly till pupa and in the

adult it slightly decreased. The result signifies that

heavy infestation of the fly of interest was abundant in

the earlier time, i.e. when the carcass was fresh.

Gradually adult disappeared and maggots began to

grow. The most important tool here is the growing

larvae through which the time of death can be

estimated. The developmental stages of the fly did not

vary much during the time period. The gradual increase

in the protein content of the developmental stages till

pupal stage and slight reduction on adult stage was a

clear indication that the adults feed on the sap of the

flesh (as the fly was in the culture chamber). The

carcass in the summer dries up very fast. So, possibly it

might happen that the carcass was not in the state in

which the flesh fly feed. Further, it is important in

poultry farm to feed the chick with a proteinaceous

diet, so the last instar or the pupal stage can be

recommended as a protein rich source.

5. Acknowledgement

The authors are very much grateful to the UGC for

their financial assistance and the head of the

Department of Zoology Gauhati University, Prof. Dr

Jatin Kalita and Prof. R. K. Bhola for their guidance

and help. The authors also express their heartfelt

gratitude to ZSI, Kolkatta for their help in

identification of the specimens.

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Adhikari K, Khanikor B, Sarma R, Mahanta S, Kalita J. Study on the life history and protein content of Sarcophaga ruficornis (Diptera:

Sarcophagidiae) a forensically important insect. Journal of Zoology Studies. 2016; 3(6):01-08.

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Sarcophaga ruficornis (Diptera: Sarcophagidiae) a …€¦ · · 2017-01-03The Journal of Zoology Studies Vol. 3 No. 6 2016 Journalofzoology.com Page 6 Table 3: Result of Tukey