Alien Registration- Ranger, Matilda P. (Strong, Franklin County)

https://digitalmaine.com/alien_docs/19296/thumbnail.jp

Stability of nuclear and mitochondrial reference genes in selected tissues of the Ambrosia beetle Xylosandrus germanus

SIMPLE SUMMARY: The ambrosia beetle Xylosandrus germanus (Blandford) is a destructive wood-boring insect of horticultural tree crops. A fungal mutualist is cultivated within host trees that provides the sole source of nutrition for the larvae and adults of this beetle. Female X. germanus adults use a pouch-like structure (i.e., mycangium) to maintain and transport spores of their fungal mutualist. To facilitate future studies examining gene expression of X. germanus’ mycangium, the identification of stable genes unaffected by experimental treatments is needed to provide a standard reference during gene expression studies. Selected tissue types were dissected from laboratory-reared and field-collected specimens of the ambrosia beetle X. germanus to evaluate the stability of five reference genes, namely, 28S ribosomal RNA (28S rRNA), arginine kinase (AK), carbamoyl-phosphate synthetase 2-aspartate transcarbamylase-dihydroorotase (CAD), mitochondrial cytochrome oxidase 1 (CO1), and elongation factor-1α (EF1α). The reference genes CO1 and AK were identified as primary and secondary reference genes. By contrast, EF1α was considered unsuitable for use as a reference gene during gene expression studies with X. germanus. These results will aid in normalizing the expression of target genes during studies with X. germanus. ABSTRACT: The fungus-farming ambrosia beetle Xylosandrus germanus (Blandford) uses a pouch-like structure (i.e., mycangium) to transport spores of its nutritional fungal mutualist. Our current study sought to identify reference genes necessary for future transcriptome analyses aimed at characterizing gene expression within the mycangium. Complementary DNA was synthesized using selected tissue types from laboratory-reared and field-collected X. germanus consisting of the whole body, head + thorax, deflated or inflated mycangium + scutellum, inflated mycangium, and thorax + abdomen. Quantitative reverse-transcription PCR reactions were performed using primers for 28S ribosomal RNA (28S rRNA), arginine kinase (AK), carbamoyl-phosphate synthetase 2-aspartate transcarbamylase-dihydroorotase (CAD), mitochondrial cytochrome oxidase 1 (CO1), and elongation factor-1α (EF1α). Reference gene stability was analyzed using GeNorm, NormFinder, BestKeeper, ΔCt, and a comprehensive final ranking by RefFinder. The gene CO1 was identified as the primary reference gene since it was generally ranked in first or second position among the tissue types containing the mycangium. Reference gene AK was identified as a secondary reference gene. In contrast, EF1α was generally ranked in the last or penultimate place. Identification of two stable reference genes will aid in normalizing the expression of target genes for subsequent gene expression studies of X. germanus’ mycangium

Weekly Survivorship Curves of Soybean Aphid Biotypes 1 and 4 on Insecticidal Seed-Treated Soybean

Thiamethoxam, an insecticide used in soybean seed treatments, effectively suppresses soybean aphids (Aphis glycines) Matsumura (Hemiptera: Aphididae) for a short time after planting. However, exactly when and how quickly soybean aphid populations could increase is unknown. Likewise, we lack data on virulent soybean aphid biotypes (that can overcome soybean resistance) when fed on seed-treated soybean. Determining the survival of soybean aphids over time on insecticidal seed-treated soybean is critical for improving soybean aphid management and may provide insights to manage aphid virulence to aphid resistant-soybean. In greenhouse and field experiments, aphid-susceptible soybean plants (with and without an insecticidal seed treatment) were infested at 7, 14, 21, 28, 35, and 42 days after planting (DAP). We compared aphid survival among biotypes 1 (avirulent) and 4 (virulent) and insecticide treatment 72 h after infestation. We also measured thiamethoxam concentrations in plant tissue using liquid chromatography-tandem mass spectrometry. As expected, soybean aphid survival was significantly lower on seed-treated soybean up to 35 DAP for both biotypes, which correlates with the decrease of thiamethoxam in the plant over time. Moreover, we found no significant difference between avirulent and virulent biotype survivorship on insecticidal seed-treated soybean plants, although we did find significantly greater survival for the virulent biotype compared with the avirulent biotype on untreated soybean in the field. In conclusion, our study further characterized the relative short duration of seed treatment effectiveness on soybean aphid and showed that survivorship of virulent aphids on seed-treated soybean is similar to avirulent aphids

Living Coast: Understanding Local Perspectives and Values of the Coast using Community Voice Method in Portsmouth and on the Durham Heritage Coast

This project was commissioned by Natural England with the aim of using the Community Voice Method to explore the values and perceptions of people in Portsmouth and along the Durham Heritage Coast in relation to their local coastal environments and the England Coast Path. The study focused on less affluent areas because wider evidence, including Natural England’s Monitor of Engagement with the Natural Environment, suggests this is where people are less likely to experience and, therefore benefit from, the natural environment. Through understanding what people think and feel about the coast, it is hoped the benefits of the England Coast Path and a healthy environment can be harnessed for a diverse range of users. The work was undertaken by the Marine Conservation Society, the University of Greenwich and Community Voice Consulting. The approach involved conducting filmed interviews with sample populations in both areas. Forty-one people were interviewed in 33 interviews across the two areas (19 in Portsmouth and 22 along the Durham Coast). The aim was to have a diverse sample of interviewees that included both regular coastal users and those who visited less often, or not at all. The sample populations were fairly evenly divided between active and limited users with a range of ages represented, although the greatest number of responses was in the 60-69 age range. Two films (each approximately 20 mins long) were produced based on the analysis of the interview transcripts. The Portsmouth CVM film can be viewed at: https://youtu.be/jJB94V2g7LY The Durham Heritage Coast CVM film can be viewed at: https://www.youtube.com/watch?v=CmCdqdLoVww A short documentary film explaining the background and method used in this research is available at: https://www.youtube.com/watch?v=Rxj_GQyCX2s The CVM films were shown in 5 locations along the Durham coast and 3 in Portsmouth in a variety of community settings. The screenings and workshops provided opportunities to involve some additional people from the local communities, add content that people thought was missing in the films, or expand topics that had already been raised. For analysis, the interviews were divided into themes related to wellbeing, place values, features of coastal places and responses to the England Coast Path

The behaviour of lubricated EHD contacts subjected to vibrations

Machine components containing contacts working in elastohydrodynamic (EHD) conditions are often subjected to vibrations. These may be originated from the mechanism or machine the contact is part of, the surrounding environment and within the contact itself. The influence of vibrations upon the behaviour of elastohydrodynamic films has been studied experimentally in a number of papers, but a comprehensive study of the effect of the parameters of the oscillatory motion upon the film thickness has not been carried out yet. In this study the authors evaluate the effect of the frequency of the oscillatory motion upon the EHD film thickness. Optical interferometry is used to measure lubricant film thickness in a ball-on-flat disc arrangement. A high – speed camera records the interferometric images for later analysis and conversion into film thickness maps. The disc runs at a constant angular velocity while the ball is driven by the traction forces developed in the EHD film. In steady state conditions, this would ensure pure rolling conditions, however in the present investigation the ball is subjected to harmonic vibrations in a direction perpendicular to the plane of the film. The contact under study is lubricated by basic oils and the temperature is kept at a constant value of 60°C. The aim of this paper is to understand how vibrations influence the lubricant film formation

Genetic Diversity of PCR-Positive, Culture-Negative and Culture-Positive Mycobacterium ulcerans Isolated from Buruli Ulcer Patients in Ghana.

Culture of Mycobacterium ulcerans from Buruli ulcer patients has very low sensitivity. Thus confirmation of M. ulcerans infection is primarily based on PCR directed against IS2404. In this study we compare the genotypes obtained by variable number of tandem repeat analysis of DNA from IS2404-PCR positive cultures with that obtained from IS2404 positive, culture-negative tissue. A significantly greater genetic heterogeneity was found among culture-negative samples compared with that found in cultured strains but a single genotype is over-represented in both sample sets. This study provides evidence that both the focal location of bacteria in a lesion as well as differences in the ability to culture a particular genotype may underlie the low sensitivity of culture. Though preliminary, data from this work also suggests that mycobacteria previously associated with fish disease (M. pseudoshottsii) may be pathogenic for humans