## 5 Harvesting of Antimicrobial Peptides from _Hermentia illucens_
### Storage conditions:  
After the inoculation of the microorganism, the larvae are incubated under desired conditions. Storage conditions include temperature, humidity and time duration. The slight modification in haemostasis of a living entity can enhance its resistivity against stressors [65]. When homothermic animals are infected by a microorganism, their body temperature increases to reduce the microbial development and stimulate the immune response of the infected host. While insects are Poikalothermic animals, which are incapable of increasing their body temperature when infected, they move to a place with a higher temperature. Additionally, it has been reported that heat shock enhances the metabolism and speeds up the synthesis of defence molecules [66]. Hence, it can be recommended to store the treated larvae at a higher temperature to obtain better results in AMP harvesting. Sample collection:  
For AMP extraction from larvae, either haemolymph is collected, or the whole larva body is utilised. [PERSON] and [PERSON] collected the haemolymph in a precooled tube consisting \begin{table}
\begin{tabular}{c c c c c c} \hline \hline
**S.No.** & **Source** & **Peptide** & **Harvesting Technique** &
\begin{tabular}{c} **Microorganisms** \\ **Inhibited** \\ \end{tabular} & **References** \\ \hline
1. & Haemolymph & & Solid phase extraction & Helicobacter pylori & [58] \\
2. & Crushed larva & stomoxynZH1 & RNA extraction & _S. aureus, E. coli,_ & \\  & & & using Trizol & _Rhizoctonia solani_ and & [59] \\
3. & Grounded larvae & & Maceration & _E. coli, P. fluorescens, M. luteus_ and _B. subtilis_ & [32] \\
4. & Larvae & & Directly used as feed & _Lactobacillus, D-steptocosci_ & [60] \\
5. & Lyophilized larvae & & Homogenised and & _Methicillin resistant_ & [61] \\  & & & extracted with & _Staphylococcus aureus_ & [62] \\
6. & Haemolymph & \begin{tabular}{c} cccropin-like \\ peptide 1 \\ \end{tabular} & \
\begin{tabular}{c} Solid-phase extraction \\ and reverse-phase \\ chromatography \\ \end{tabular} &
\begin{tabular}{c} _E. coli, Enterobacter_ \\ _aerogens_ and \\ _Pseudomonas aeruginosa_ \\ \end{tabular} & [62] \\
7. & Grounded larva & & &
\begin{tabular}{c} Maceration \\ using methanol \\ \end{tabular} & _Salmonella_ and _E. coli_ & [63] \\
8. & Larvae whose digestive & \begin{tabular}{c} Hidefensin-1, \\ Hidiptericin-1 \\ and HiCG13551 \\ \end{tabular} & \
\begin{tabular}{c} TRIeasy- RNA \\ isolation kit \\ \end{tabular} &
\begin{tabular}{c} _Streptococcus pneumonia,_ \\ _E. coli_ and \\ _Staphylococcus aureus_ \\ \end{tabular} & [64] \\ \hline \hline \end{tabular}
\end{table} 
Table 3: Antimicrobial activity of peptides extracted from _H. illucens_. of protease and melanisation inhibitors. Additionally, for small-sized insects, they froze the insects using liquid nitrogen, followed by grinding. Then, the powder was placed in water acidified with trifluoroacetic acid embodied with protease and melanisation inhibitor, followed by centrifugation and filtration [67]. Similarly, in most of the research, it was found that haemolymph is collected in ice-cold tubes consisting of phenylthiourea crystals to avoid coagulation. After collection, haemolymph is centrifuged to remove haemocytes [51]. [PERSON] proposed a method for collecting haemolymph from larva using a collection tube. This tube was prepared by making a hole at the bottom of a 0.5-mL centrifuge tube and then placing this tube in a 1.5 mL tube [68]. Samples were collected from grounded larvae in a solution of methanol, water and acetic acid for characterisation. Additionally, the method to isolate RNA was considered as follows: dissect larvae in ice-cold PBS, soak foregut/ midgut, hindgut and salivary glands in lysis buffer, and then freeze [32]. Another protocol was reported in which larvae were ground and suspended in 20% acetic acid solution, followed by boiling and then centrifugation [55].