The aim of this Special Issue reprint is to introduce the reader to the latest biotechnological approaches used in crop protection by employing Bacillus thuringiensis (Bt) and its invertebrate active proteins. This Special Issue brought together descriptions of novel isolated Bt strains and insecticidal proteins with previously undescribed biocidal activities, reflecting the ongoing quest for innovation in crop protection to overcome insect resistance and expanding host ranges. Altogether, the content of this reprint will offer multiple benefits for users in agribusiness, the economy, and wider society.

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