Tephritidae fruit flies (Diptera: Tephritidae) are regarded as important damage-causing species due to their ability to cause great economic losses in fruit and vegetable crops. Bactrocera minax and Bactrocera tsuneonis are two sibling species of the subgenus Tetradacus of Bactrocera that are distributed across a limited area of China, but have caused serious impacts. They share similar morphological characteristics. These characteristics can only be observed in the female adult individuals. The differences between them cannot be observed in preimaginal stages. Thus, it is difficult to distinguish them in preimaginal stages morphologically. In this study, we used molecular diagnostic methods based on cytochrome c oxidase subunit I and species-specific markers to identify these two species and improve upon the false-positive results of previous species-detection primers. DNA barcode sequences were obtained from 900 individuals of B. minax and 63 individuals of B. tsuneonis. Based on these 658 bp DNA barcode sequences of the cytochrome c oxidase subunit I gene, we successfully designed the species-specific primers for B. minax and B. tsuneonis. The size of the B. minax specific fragment was 422 bp and the size of the B. tsuneonis specific fragment was 456 bp. A series of PCR trials ensured the specificity of these two pairs of primers. Sensitivity assay results demonstrated that the detection limit for the DNA template concentration was 0.1~1 ng/µL for these two species. In this study, we established a more reliable, rapid, and low-cost molecular identification method for all life stages of B. minax and B. tsuneonis. Species-specific PCR can be applied in plant quarantine, monitoring and control of B. minax and B. tsuneonis.
Worldwide bike-sharing systems are growing in popularity as an alternative, environmentally friendly mode of transportation. As cities seek to further develop bike-sharing programmes, it is important to consider how systems should expand to simultaneously address existing inequalities in accessibility, and best serve demand. In this paper, we determine ideal locations for future bikesharing stations in Glasgow, Scotland, by integrating demand modelling with accessibility considerations. We began by analysing the spatio-temporal trends of bike-sharing usage, and assessed the spatial equity of access to stations in Glasgow. To identify important determinants of bikesharing demand, we ran an ordinary least squares regression model using bike sharing trip data from Nextbike Glasgow. We then quantifiably measured the level of spatial accessibility to stations by applying the two-step floating catchment area (2SFCA) methodology and ran a GIS weighted overlay analysis using the significant determinants of station demand. Lastly, we combined the demand and accessibility results to determine where new stations should be located using a maximum covering location problem (MCLP) that maximized the population served. Our results show that distance from transit stations, distance from downtown, employment rates, and nearby cycling lanes are significant factors affecting station-level demand. Furthermore, levels of spatial access were found to be highest primarily in the centre and eastern neighbourhood of Glasgow. These findings aided in determining areas to prioritize for future station locations, and our methodology can easily be applied to other bike-share programmes with adjustments according to varying aims for system expansion.
Bactrocera dorsalis (Hendel) is a notorious agricultural pest worldwide, and its prevention and control have been widely studied. Bacteria in the midgut of B. dorsalis help improve host insecticide resistance and environmental adaption, regulate growth and development, and affect male mating selection, among other functions. Insects have an effective gut defense system that maintains self-immunity and the balance among microorganisms in the gut, in addition to stabilizing the diversity among the gut symbiotic bacteria. However, the detailed regulatory mechanisms governing the gut bacteria and self-immunity are still unclear in oriental fruit flies. In this study, the diversity of the gut symbiotic bacteria in B. dorsalis was altered by feeding host fruit flies antibiotics, and the function of the gut bacteria was predicted. Then, a database of the intestinal transcriptome of the host fruit fly was established and analyzed using the Illumina HiSeq Platform. The gut bacteria shifted from Gram negative to Gram positive after antibiotic feeding. Antibiotics lead to a reduction in gut bacteria, particularly Gram-positive bacteria, which ultimately reduced the reproduction of the host flies. Ten immunity-related genes that were differentially expressed in the response to intestinal bacterial community changes were selected for qRT-PCR validation. Peptidoglycan-recognition protein SC2 gene (PGRP-SC2) was one of the 10 immunity-related genes analyzed. The differential expression of PGRP-SC2 was the most significant, which confirms that PGRP-SC2 may affect immunity of B. dorsalis toward gut bacteria.
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