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Research
Development of Sarcophaga dux (Diptera: Sarcophagidae) at constant temperatures and differential gene expression for age estimation of intrapuparial
Yadong Guo
Department of Forensic Science,School of Basic Medical Science,Central South University,Changsha,Huan,China
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Sarcophaga dux (Diptera: Sarcophagidae) is a necrophagous flesh fly with potential forensic value in estimating minimum postmortem interval (PMImin). The basic developmental data and precise intrapuparial age estimates are significant for PMImin estimationof application entomological data in legal medicine.
Methods
The development parameters of S. dux at seven constant temperatures from 16°C to 34°C were investigated by rearing using pig lung in the artificial climate box. The appropriate reference genes and intrapuparial differentially expressed genes (DEGs) of S. dux at constant temperatures 34, 25 and 16°C were selected and analyzed using RT-qPCR for more precisely age estimations.
Results
The developmental durations of S. dux at 16, 19, 22, 25, 28, 31 and 34°C from larviposition to adult eclosion were 1478.6±18.3, 726.1±15.8, 538.5±0.9, 394.1±9.5, 375.6±10.8, 284.1±7.3, and 252.5±6.1 h, respectively. The thermal summation constant of S. dux was 5341.71±249.29 degree hours, and the developmental threshold temperature was 12.266±0.35°C. The most reliable reference genes under intrapuparial and different temperature in our study were: GST1 and 18S rRNA for 34°C group, GST1 and RPL49 for 25 °C group, and 18S rRNA and 28S rRNA for 16 °C group，the four differential expression genes (Hsp60, A-alpha, ARP, RPL8) can be used to more precisely intrapuparial age estimation of S. dux.
Conclusions
The basic developmental data of S. dux at constant temperatures, such as body length changing of the larva period, accumulated degree hours and duration of development, to establish developmental models that can be used to estimate the PMImin. The selection and evaluation of appropriate reference genes at different experimental conditions are primary with RT-qPCR. The differentially expressed gene can contribute to more accuracy age estimations of S. dux intrapuparial. The result from this study can make contributes to the use of S.dux for estimating PMImin.
Keywords
Postmortem interval, Forensic entomology, developmental datas, differentially expressed genes, Sarcophaga dux
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This is a preprint. It has not completed peer review.
Research
Development of Sarcophaga dux (Diptera: Sarcophagidae) at constant temperatures and differential gene expression for age estimation of intrapuparial
Yadong Guo
Department of Forensic Science,School of Basic Medical Science,Central South University,Changsha,Huan,China
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 31 Dec, 2019
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Parasitology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Sarcophaga dux (Diptera: Sarcophagidae) is a necrophagous flesh fly with potential forensic value in estimating minimum postmortem interval (PMImin). The basic developmental data and precise intrapuparial age estimates are significant for PMImin estimationof application entomological data in legal medicine.
Methods
The development parameters of S. dux at seven constant temperatures from 16°C to 34°C were investigated by rearing using pig lung in the artificial climate box. The appropriate reference genes and intrapuparial differentially expressed genes (DEGs) of S. dux at constant temperatures 34, 25 and 16°C were selected and analyzed using RT-qPCR for more precisely age estimations.
Results
The developmental durations of S. dux at 16, 19, 22, 25, 28, 31 and 34°C from larviposition to adult eclosion were 1478.6±18.3, 726.1±15.8, 538.5±0.9, 394.1±9.5, 375.6±10.8, 284.1±7.3, and 252.5±6.1 h, respectively. The thermal summation constant of S. dux was 5341.71±249.29 degree hours, and the developmental threshold temperature was 12.266±0.35°C. The most reliable reference genes under intrapuparial and different temperature in our study were: GST1 and 18S rRNA for 34°C group, GST1 and RPL49 for 25 °C group, and 18S rRNA and 28S rRNA for 16 °C group，the four differential expression genes (Hsp60, A-alpha, ARP, RPL8) can be used to more precisely intrapuparial age estimation of S. dux.
Conclusions
The basic developmental data of S. dux at constant temperatures, such as body length changing of the larva period, accumulated degree hours and duration of development, to establish developmental models that can be used to estimate the PMImin. The selection and evaluation of appropriate reference genes at different experimental conditions are primary with RT-qPCR. The differentially expressed gene can contribute to more accuracy age estimations of S. dux intrapuparial. The result from this study can make contributes to the use of S.dux for estimating PMImin.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8