Abstract

Using insect hemolymph (“blood”) and insect body surface elutions, researchers can perform rapid and cheap biochemical analyses to determine the insect’s immunology status. The authors of this publication describe a detailed methodology for a quick marking of the concentration of total proteins and evaluation of the proteolytic system activity (acid, neutral, and alkaline proteases and protease inhibitors), as well as a methodology for quick “liver” tests in insects: alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and urea and glucose concentration analyses. The meaning and examples of an interpretation of the results of the presented methodology for biochemical parameter determination are described for the example of honey bees.

Highlights

  • Insects are the largest group of animals

  • The value protein concentration in the analysis sample is for further analysis of protein concentration in the of sample is necessary for further ofnecessary the proteolytic system

  • Carrying out rapid biochemical tests to determine the activity of the proteolytic system makes sense for the hemolymph solution and the elution from the body surface, which is not the case with rapid “liver” analyses and concentrations of urea and glucose, as shown in

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Summary

Introduction

Insects are the largest group of animals. They occur in almost all types of environment around the world. Sick and weakened insects can be a source of infection for other invertebrates; they sometimes cause huge economic losses (it is estimated that the value of pollination by honey bees in one year in the United States is worth about 14.6 billion USD) [3]. They may cause initially invisible changes that negatively affect other larger organisms, including humans [4]. An additional advantage and argument for conducting screening biochemical tests on invertebrates is that they are often treated as lower-level animals; the research using them does not require applying for legal permits and is not restricted in most countries

Materials
Hemolymph Collection
Insect immobilization
Elution from the Insect’s Body Surface
Total Protein
The value shown in Figure
Protease Activity Analyses
The optimal pH have should be added
Protease Activity Level Calculation
Protease Inhibitor Activity Analyses
Calculation of Protease Inhibitor Activity Levels
Aspartate
Calculation
Urea Concentration
Glucose
Analysis Adjusted to the Type of Biological Material
Additional Tips for Applying Presented Methodologies
Meanings of the Proteolytic System Activity
The Proteolytic System in the Hemolymph
Urea and Glucose Parameters
Conclusions
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