A Sustainable Technique for Colony Multiplication by Eduction of Wild Nests of the Stingless Bee Tetragonula iridipennis Smith

Colony multiplication of stingless bees, Tetragonula iridipennis, largely relies on the eduction of wild colonies from their natural nesting sites in India. During the hiving of wild colonies, colonies were destroyed with the loss of robust wild foragers and built-in storage reserves over the years. The present study was conducted to devise a technique to sustainably multiply the colonies of stingless bees from the wild colony and the colony establishment and development during the eduction process. The annexure hives provided for eduction were accepted in a shorter time (3.25 ± 1.18 days), with the construction of storage pots observed at 7.75 ± 1.59 days after hive acceptance by the bees. The movement of foragers between the wild colony and the annexure hives was noticed for 13.80 ± 4.20 days. The foragers settled in the annexure hives and started foraging after 18.20 ± 2.49 days. The advancing fronts were observed at 26.67 ± 2.58 days after the addition of the laying queen in the established annexure hives. There was a significant increase in the number of inhive workers after the queen seeding in the annexure hives. This technique is the easiest and most sustainable non- destructive way of multiplication of stingless bee colonies without loss in viability of the perennial wild colony

Development of recombinase polymerase amplification-based colorimetric detection assay for rapid identification of invasive cassava mealybug, Phenacoccus manihoti Matile-Ferrero

Phenacoccus manihoti Matile-Ferrero (Hemiptera: Pseudococcidae), is an economically important invasive cassava pest responsible for the massive devastation of cassava in Asia and African continent. Initially, identifying this invasive pest posed challenges because it closely resembled native mealybug species. Additionally, the traditional morphological identification process is labor-intensive and time-consuming. Detecting invasive pests at an early stage is crucial, hence development of a rapid detection assay is essential. In the current study, we have developed a simple, rapid, sensitive, and efficient molecular detection assay for P. manihoti based on Recombinase Polymerase Amplification (RPA). The primers for the RPA assay were designed using unique nucleic acid sequences of P. manihoti, and the protocol was standardized. Specificity test demonstrated that the RPA assay could amplify DNA of P. manihoti only, and no amplification was observed in six other mealybug species. The specificity of assay was confirmed using SYBR green-based colorimetric detection and gel electrophoresis where positive samples showed 195 bp amplicon size in P. manihoti samples. The assay successfully amplified P. manihoti DNA in thirty minutes at an annealing temperature of 41° C in a water bath and displayed a sensitivity of 72.5 picograms per microliter. The assay's simplicity, rapidity, and high sensitivity make it a valuable tool for detecting and monitoring P. manihoti in quarantine stations and facilitating in development of a portable diagnostic kit