Effects of Codeine, Sodium Pentothal and Different Temperature Factors on the Growth Rate Development of Chrysomya rufifacies for the Forensic Entomotoxicological Purposes

Forensic Entomology is the use of insects and other arthropods in forensic investigation concerning decomposed bodies and it has become the “gold standard” for estimating time since death in many countries. In addition to estimating the post-mortem interval (PMI) insects that feed on carcasses may also represent a reliable specimen for toxicological analyses (Entomotoxicology) [1-3]. It is very useful for cases where the body has been long dead. Different Species of insects lay eggs on dead compost, Forensic entomologists done research on this kind of insects and their larval lifecycles finally they determine the body has been dead before three days or four days ago. After three days of investigation, insect evidence is most accurate in some method of determining duration time since death. Recently, I have also analyzed this kind of cases’ in which duration time since death was only a few hours previous to discovery [4-6].


Introduction
Forensic Entomology is the use of insects and other arthropods in forensic investigation concerning decomposed bodies and it has become the "gold standard" for estimating time since death in many countries. In addition to estimating the post-mortem interval (PMI) insects that feed on carcasses may also represent a reliable specimen for toxicological analyses (Entomotoxicology) [1][2][3]. It is very useful for cases where the body has been long dead. Different Species of insects lay eggs on dead compost, Forensic entomologists done research on this kind of insects and their larval lifecycles finally they determine the body has been dead before three days or four days ago. After three days of investigation, insect evidence is most accurate in some method of determining duration time since death. Recently, I have also analyzed this kind of cases' in which duration time since death was only a few hours previous to discovery [4][5][6].
Two main ways of using insects to determine duration time since death, by using succession ally waves of insects, using maggot age and its development in three different methods as follows, The first method is used when the corpse has been dead for between a month up to a year or more, and the second method is used when death occurred less than a month prior to discovery [7][8][9][10].
1. These samples were collected randomly from the meat shops.
Meat kept in open environment in Noida and was subjected for collection.
2. The sample flies collected were subjected for collection and rearing of flies.
4. 50 flies were used in this study, placed in 12 jars (4 each). These flies were allowed to rear under different environmental conditions and different drugs ethanol, and cannabis.
5. Vermiculite was filled in rearing chamber.
6. 12 jars placed to observe the colonization of the blow flies.
7. Meats were placed inside the jars treated with drugs.

Control sample
Condition: Room temperature (Dry): The jar containing adult blow flies were placed at room temperature on the 11 th march. The eggs were observed to have been laid by the 14 th march. On the 3 rd day after incubation the 1 st in star stage was observed. From which point counting was performed after 6 hour. By the 78 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 90 hours was of the pupa (Tables 1 and 2).

Control sample
Condition: Room temperature (Humid): The jar containing adult blow flies were placed at room temperature on the 26 th march. On the 4 th day after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 78 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 84 hours was of the pupa (Tables  3 and 4).

Control sample
Condition: Room temperature (Dry): The jar containing adult blow flies were placed at room temperature on the 26 th march. The eggs were observed to have been laid by the 30 th march. On the 6 th day after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 90 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 102 hours was of the pupa (Tables 5 and 6).

Control sample
Condition: Room temperature (Humid): The jar containing adult blow flies were placed at room temperature on the 26 th march. On the 6 th day after incubation the 1 st instar stage of larvae were observed. The 1 st instar stages of larvae were first observed on the 31 th march. From which point counting was performed after 6 hour. By the 84 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 96 hours was of the pupa (Tables 7 and 8).

Ethanol treated sample
Condition: Room temperature (Dry): The jar containing adult blow flies were placed at room temperature on the 15 th march. The eggs were observed to have been laid by the 17 th march. On the 4 th day (18 th march) after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 66 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 72 hours was of the pupa (Tables 9 and 10).

Ethanol treated sample
Condition: Room temperature (Humid): The jar containing adult blow flies were placed at room temperature on the 26 th march. The eggs were observed to have been laid by the 28 th march. On the 5 th day (30 th march) after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 54 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 66 hours (2 nd April) was of the pupa (Tables 11 and 12).     flies were placed at room temperature on the 26 th march. On the 4 th day (29 th march) after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 66 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 84 hours was of the pupa (Tables 13 and 14).

Ethanol treated sample
Condition: Cool temperature (Humid): The jar containing adult blow flies were placed at room temperature on the 26 th march. The eggs were observed to have been laid by the 29 th march. On the 5 th day (30 th march) after incubation, the 1 st in star stage of larvae were observed. From which point counting was performed after 6 hour. By the 54 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 66 hours was of the pupa (Tables 15 and 16).

Cannabis treated sample
Condition: Room temperature (Dry): The jar containing adult blow flies were placed at room temperature on the 15 th march. The eggs were observed to have been laid by the 17 th march. On the 4 th day (18 th march) after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 72 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 78 th hours was of the pupa (Tables 17 and 18).

Cannabis treated sample
Condition: Room temperature (Humid): The jar containing adult blow flies were placed at room temperature on the 26 th march. The eggs were observed to have been laid by the 28 th march. On the 4 th day (29 th march) after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 60 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 72 hours was of the pupa (Tables 19 and 20).

Cannabis treated sample
Condition: Cool temperature (Dry): The jar containing adult blow flies were placed at room temperature on the 26 th march. The eggs were observed to have been laid by the 29 th march. On the 4 th day (30 th march) after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 72 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 84 hours was of the pupa (Tables 21 and 22).

Cannabis treated sample
Condition: Cool temperature (Humid): The jar containing adult blow flies were placed at room temperature on the 2 nd April. The eggs were observed to have been laid by the 5 th April. On the 5 th day (6 th April) after incubation the 1 st instar stage of larvae were observed. From which point counting was performed after 6 hour. By the 66 th hour, the counting was made of a mixture of a larva and pupa and the final reading taken at 72 hours was of the pupa (Tables 23 and 24).

Discussion and Conclusion
In the above study the growth and colonization of blow flies of species Chrysomya rufifacies were studied under different conditions. On the basis of result it was clearly seen that a changes in temperature and humidity bring about a significant changes in growth pattern of the larval stages.
In the condition with the higher temperature larva developed quickly and matured into pupa when compared to the sample grown in cooler temperature. It was also noted that fly larva grew and mature faster when they were placed under humid conditions [15][16][17].
When the effects of the toxins on the growth rates were observed, a clearly distinct change was seen in the growth pattern. Where the control sample took an average of 4 days to grow from 1 st instar to pupae stages, the samples grown in the presence of ethanol and cannabis showed a much faster growth rates.     The number of larvae observed also showed significant differences with the maximum reproduction occurring with the control sample, followed by the cannabis and ethanol showing the least number of larvae [18][19][20][21][22][23].
Therefore it can be concluded that both the studies that were put forward before the start of this study have been proven and that the differences in environmental conditions and presence of drugs affect the growth and colonization of blow flies [24,25]. This study demonstrates again the necessity of considering the possible effects of drugs in tissues on insect growth rates when estimating the postmortem interval (PMI) using entomological techniques.