Abstract

SUMMARYResearch backgroundThe presence of insect fragments is one of the major constrains in stored food commodities and it causes considerable loss in the quality of the produce. The management of the pest is viewed as a huge challenge in foodprocessingindustry. Conventionally, the detection of Tribolium castenaum in the food processing industry is carried out by acid hydrolysis and staining methods that are time consuming and lack precision.Experimental approachConsidering the importance of a quick and effective method, a quantitative polymerase chain reaction (qPCR)-based approach was developed and elucidated in this study. The mitochondrial cytochrome oxidase I (mtCOI) gene was identified as a target due to its abundance in the pest. Specific primers were designed against the target gene by Primer Premier software and amplified in a qPCR.Results and conclusionsThis method is capable of detecting all the ontogenic stages of T. castaneum in stored wheat flour. Earlier experiments had demonstrated that about 20 µg of DNA can be obtained from 2.2 mg of insects. To quantify the infestation levels, the cycle threshold (Ct) values obtained from known samples were subjected to regression analysis and expressed as adult equivalents. In the unknown samples, the infestation was calculated as 1.74 and 0.046 adult insects in 5 g of wheat flour. The maximum permissible limit of insect fragments in flour is 75 insect fragments or approx. 3 adults per 50 g of flour as per the US Food and Drug Administration (FDA). Hence, by adopting this new method, it is possible for the warehouse operators to arrive at a decision to proceed with efficient management practices where wheat flour is stored. Also, this method can be ratified by government agencies associated with international business to ascertain whether the wheat flour meets the standards set by the respective country before subjecting to foreign trade.Novelty and scientific contributionThis study is the first of its kind in the detection and quantification of T. castaneum in milled products. So far, only conventional methods have been employed to assess the presence of the pests and manual counting of fragments are practiced to quantify the infestation levels. The developed qPCR method is faster, reliable and can be employed in milling industries, bakery industries, food processing plants and foreign trade units for critical detection and quantification of T. castaneum pest infestation.

Highlights

  • Wheat is one of the major cereal crops cultivated in India and its production has reached an all-time high of 99.70 million tonnes in 2019 [1]

  • This method is capable of detecting all the ontogenic stages of T. castaneum in stored wheat flour

  • Primer specificity and multiple sequence alignment The main aim of this study was to develop a molecular technique to detect and quantify the presence of red flour beetle in wheat flour. quantitative real-time polymerase chain reaction (qRT-PCR) is a well-known technique based on gene expression for rapid detection and quantification of T. castaneum in food samples

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Summary

Introduction

Wheat is one of the major cereal crops cultivated in India and its production has reached an all-time high of 99.70 million tonnes in 2019 [1]. India ranks second in the production of wheat, despite the total postharvest losses of about 33.5 % in stored wheat and wheat flour [2]. The major issue is the presence of insects, which is responsible for 5–15 % loss during storage [3]. Tribolium castaneum (red flour beetle) is one of the major pests infesting wheat flour during post harvest processing. Even when high standards of sanitary and handling procedures are practiced, the flour is susceptible to infestation by pests [4]. Biotechnol. 59 [1] 112–121 (2021)

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