Invasion genetics and development of rapid diagnostics of insect pests on traded plants

Background: Global trade of plant products is a major driving force for the unintended spread of economically harmful insect pests. This PhD thesis aimed at (i) developing and implementing molecular tools for the on-site identification of invasive insect pests at points of entry (POEs) for plant import products as a prevention measure; and (ii) investigating the invasion history of the mosaic leafhopper Orientus ishidae, a potential vector of grapevine Flavescence dorée phytoplasma. Methods: To achieve the first goal, loop-mediated isothermal amplification (LAMP)-based genetic assays for the rapid on-site identification of Bemisia tabaci, Thrips palmi and several invasive fruit flies of the genera Bactrocera and Zeugodacus were developed. Using publicly available DNA sequences, LAMP primers were designed to specifically target a fragment of the mitochondrial cytochrome c oxidase subunit 1 gene. To address the second goal of this PhD thesis, the invasion genetics of O. ishidae was studied, an invasive insect species that spread from its native range from in East Asia to North America in the first half of the 20th century and only recently colonised Europe. Possible source populations and invasion pathways were investigated by assessing the genetic structure of 41 O. ishidae populations from Asia, Europe, and North America based on a mitochondrial marker and 641 single nucleotide polymorphisms (SNPs) generated by double digest restriction-site associated DNA (ddRAD) sequencing. Results: Validation performed under laboratory and on-site conditions demonstrated the robustness and reliability of the developed LAMP identification assays. Analysing 319 insect specimens, the overall diagnostic test efficiency was 98% and the overall diagnostic test specificity was 100%. The small number of false-negative results (2%) originated either from previously unknown biotypes, not included in the initial primer design, or from handling errors during LAMP preparation. The results from the molecular genetic analyses of O. ishidae revealed a clear genetic separation between a native population from Asia and the non-native populations from Europe and North America. Among the non-native populations, only faint signals of spatial genetic structuring were found. However, when comparing non-native populations from Europe and North America, elevated levels of admixture of genetically distant mitochondrial haplotypes were observed for European populations. Conclusion: Characterised by high analysis speed (<1 h) and simplicity in use (only 1 pipetting step), the validated LAMP assays were found to be suitable identification tools for on-site application by plant health inspectors. Since completion of the validation phase, the developed identification assays are routinely deployed in the phytosanitary import control process of Switzerland. The considerable genetic separation between native and non-native populations of O. ishidae together with the strikingly high genetic similarity of European and North American populations suggest an invasion scenario in which North American populations served as source for the European invasion. A slightly reduced genetic structure combined with increased admixture of genetically distant mitochondrial haplotypes furthermore indicate that the European colonisation history was shaped by multiple introductions from North America, complemented by frequent intra-European gene flow. Taken together, it is hypothesised that the overall genetic complexity of non-native populations was strongly driven by frequent international trade of plants infested by O. ishidae

Réduction des risques et jeux d'argent. Questions ouvertes par la révision du dispositif suisse

Initialement associé à l’approche de santé publique des drogues illégales, le concept de réduction des risques et dommages (RDR) a été défini par des programmes et des pratiques visant à réduire les conséquences négatives des conduites addictives sur les plans sanitaire, social et économique, et ce, sur la base d’indicateurs scientifiquement pertinents. En matière de jeu excessif, l’approche de santé publique demeure le parent pauvre et le concept de RDR n’a été implanté que de manière marginale, subordonnée à des politiques soucieuses de maximiser les recettes de l’État. En Suisse, la révision de la législation sur les jeux d’argent réactualise la question de la RDR appliquée au jeu excessif. En érigeant la protection des joueurs au rang d’objectif premier du dispositif projeté, le gouvernement suisse a contraint ses réseaux de prévention à revenir sur les fondements de la RDR, à considérer l’efficience des différentes mesures de prévention du jeu excessif et à faire un bilan critique du dispositif de régulation préexistant. Si l’addiction aux jeux d’argent est aujourd’hui reconnue comme une maladie, il y a lieu d’interroger la responsabilité de l’État sous l’angle du droit à la santé. Cependant, il existe un conflit structurel entre la perception de la santé publique et la perception des opérateurs de jeu, qui ont des impératifs économiques de rentabilité. Pour la santé publique, la liberté économique des opérateurs est, par définition, restreinte par l’intervention de l’État, car les opérateurs remplissent la fonction d’auxiliaires chargés d’offrir un jeu à moindre risque par des monopoles d’État ou des régimes d’octroi de licences. Pour les opérateurs, il s’agit d’abord de préserver la liberté économique. Un débat sur les nouveaux cadres de régulation et une redéfinition des objectifs des politiques publiques concernant les jeux d’argent apparaissent inévitables.The concept of Harm Reduction (HR) was initially associated with the public health approach towards illegal drugs. This concept has been defined as programmes and practices to reduce the negative consequences of addictive drives upon health, social and economic functioning. Such reductions are based upon observations of scientifically relevant indicators. Regarding excessive gambling, the public health approach remains largely overlooked. The concept of RR has only been implemented in a marginal way, subordinate to policies concerned with maximizing the State’s income. In Switzerland, the revision of gambling legislation once again raises the question of applying HR to excessive gambling. By placing the protection of players as a projected primary objective, the Swiss government forces its prevention networks to return to HR foundation, to draw attention on the effectiveness of different prevention measures for excessive gambling, and to make a critical evaluation of pre-existing regulation practices. Today gambling addiction is recognized as an illness, and in doing so, an obligation for the State as a right to health issue. However, there is an existing structural conflict between the public health perspective and that of gambling operators, whose economic aim is to make profit. For public health, the economic freedom of operators is, by definition, restricted by the intervention of the State. The operators fulfill the role of auxiliaries, charged with the responsibility of providing a game with less risk, by State monopolies or grant licensing regimes. For the operators, such a vision hinders their hopes of economic freedom. A debate on the new regulating framework and a redefinition of the objectives of public policies towards gambling appears inevitable.Relacionado en un principio con el enfoque de salud pública sobre drogas ilegales, el concepto de reducción de riesgos y daños (RDR) fue definido por programas y prácticas destinados a reducir las consecuencias negativas de los comportamientos adictivos en los planos sanitario, social y económico, sobre la base de indicadores específicamente pertinentes. En materia de juego excesivo, el enfoque de salud pública continúa siendo el pariente pobre y el concepto de RDR fue implantado solamente de manera marginal, subordinado a políticas preocupadas por maximizar los ingresos del Estado. En Suiza, la revisión de la legislación sobre los juegos por dinero reactualiza la cuestión de la RDR aplicada al juego excesivo. Colocando la protección de los jugadores en el rango de primer objetivo del dispositivo proyectado, el Gobierno suizo ha obligado a sus redes de prevención a volver a considerar los fundamentos de la RDR y la eficacia de las diferentes medidas de prevención del juego excesivo y a hacer un balance crítico del dispositivo de reglamentación preexistente. Si se reconoce en la actualidad que la adicción a los juegos por dinero es una enfermedad, este hecho permite interrogar la responsabilidad del Estado desde el punto de vista del derecho a la salud. Sin embargo, existe un conflicto estructural entre la percepción de la salud pública y la percepción de los operadores de juego, que tienen imperativos económicos de rentabilidad. Para la salud pública, la libertad económica de estos operadores está, por definición, limitada por la intervención del Estado, puesto que los mismos cumplen la función de auxiliares, encargados de ofrecer un juego con menos riesgos para los monopolios del Estado o los regímenes de otorgamiento de licencias. Para los operadores, se trata en primer lugar de preservar la libertad económica. Parece inevitable en este contexto la necesidad de mantener un debate sobre los nuevos marcos regulatorios y la redefinición de los objetivos de las políticas públicas en lo que respecta a los juegos por dinero

Mitochondrial plasticity in brachiopod ( Liothyrella spp.) smooth adductor muscle as a result of season and latitude

Habitat temperature and mitochondrial volume density (Vv(mt,mf)) are negatively correlated in fishes, while seasonal acclimatization may increase Vv(mt,mf) or the surface density of the mitochondrial cristae (Sv(im,mt)). The effect of temperature on invertebrate mitochondria is essentially unknown. A comparison of two articulate brachiopod species, Liothyrella uva collected from Rothera Station, Antarctica in summer 2007, and Liothyrella neozelanica collected from Fiordland, New Zealand in winter 2007 and summer 2008, revealed a higher Vv(mt,mf) in the Antarctic brachiopod. The Sv(im,mt) was, however, significantly lower, indicating the Antarctic brachiopods have more, less reactive mitochondria. L. uva, from the colder environment, had larger adductor muscles in both absolute and relative terms than the temperate L. neozelanica. Furthermore, a seasonal comparison (winter vs. summer) in L. neozelanica showed that the absolute and relative size of the adductor increased in winter, Vv(mt,mf) was unchanged, and Sv(im,mt) was significantly increased. Thus, seasonal acclimatization to the cold resulted in the same number of more reactive mitochondria. L. neozelanica was clearly able to adapt to seasonal changes using a different mechanism, i.e. primarily through regulation of cristae surface area as opposed to mitochondrial volume density. Furthermore, given the evolutionary age of these living fossils (i.e. approximately 550million years), this suggests that mitochondrial plasticity has roots extending far back into evolutionary histor

Risk Factors for Fear of Recurrence in Head and Neck Cancer Patients.

OBJECTIVE Fear of recurrence (FoR) affects the quality of life of head and neck cancer survivors. Identification of factors predisposing to FoR may help to recognize and treat patients at risk. MATERIALS AND METHODS For this exploratory study, 101 disease-free head and neck cancer survivors completed a cross-sectional survey in 2017 that included the FoR questionnaire at a random point in time during their follow-up. Additionally, the patients were asked to choose their favorite among four follow-up schedules with or without systematic imaging and varying frequency of visits. RESULTS Elevated FoR was present in 36.6% of patients. Females and patients ≤65 years showed significantly higher FoR overall scores than males (score difference 3.40; CI 0.49-6.32; p = 0.022) and patients >65 years (score difference 4.25; CI 1.58-6.92; p = 0.002). A history of cancer recurrence or second primary malignancy increased the relative risk (RR) for elevated FoR (RR 1.7; CI 1.01-2.86; p = 0.046). Tumor stage and treatment modality were not significantly associated with elevated FoR or FoR overall score. Higher FoR overall scores were recorded in patients who favored intensive follow-up plans (mean overall FoR score 18 vs. 15; SD 7.7; p = 0.076) and systematic imaging in follow-up (17 vs. 13, SD 7.1; p = 0.034). CONCLUSION Fear of recurrence in head and neck cancer patients is associated with female sex, younger age, and history of a past recurrence or second primary malignancy. Due to its high prevalence, it should be addressed in clinical practice and future research. LEVEL OF EVIDENCE N/A Laryngoscope, 2022

Risk Factors for Fear of Recurrence in Head and Neck Cancer Patients

Objective: Fear of recurrence (FoR) affects the quality of life of head and neck cancer survivors. Identification of factors predisposing to FoR may help to recognize and treat patients at risk. Materials and methods: For this exploratory study, 101 disease-free head and neck cancer survivors completed a cross-sectional survey in 2017 that included the FoR questionnaire at a random point in time during their follow-up. Additionally, the patients were asked to choose their favorite among four follow-up schedules with or without systematic imaging and varying frequency of visits. Results: Elevated FoR was present in 36.6% of patients. Females and patients ≤65 years showed significantly higher FoR overall scores than males (score difference 3.40; CI 0.49-6.32; p = 0.022) and patients >65 years (score difference 4.25; CI 1.58-6.92; p = 0.002). A history of cancer recurrence or second primary malignancy increased the relative risk (RR) for elevated FoR (RR 1.7; CI 1.01-2.86; p = 0.046). Tumor stage and treatment modality were not significantly associated with elevated FoR or FoR overall score. Higher FoR overall scores were recorded in patients who favored intensive follow-up plans (mean overall FoR score 18 vs. 15; SD 7.7; p = 0.076) and systematic imaging in follow-up (17 vs. 13, SD 7.1; p = 0.034). Conclusion: Fear of recurrence in head and neck cancer patients is associated with female sex, younger age, and history of a past recurrence or second primary malignancy. Due to its high prevalence, it should be addressed in clinical practice and future research

Ultrastructure of pedal muscle as a function of temperature in nacellid limpets

Temperature and mitochondrial plasticity are well studied in fishes, but little is known about this relationship in invertebrates. The effects of habitat temperature on mitochondrial ultrastructure were examined in three con-familial limpets from the Antarctic (Nacella concinna), New Zealand (Cellana ornata), and Singapore (Cellana radiata). The effects of seasonal changes in temperature were also examined in winter and summer C. ornata. Stereological methods showed that limpet pedal myocytes were 1-2 orders of magnitude smaller in diameter (≈3.5μm) than in vertebrates, and that the diameter did not vary as a function of temperature. Mitochondrial volume density (Vv(mt,f)) was approximately 2-4 times higher in N. concinna (0.024) than in the other species (0.01 and 0.006), which were not significantly different from each other. Mitochondrial cristae surface density (Sv(im,mt)) was significantly lower in summer C. ornata (24.1±0.50μm2μm−3) than both winter C. ornata (32.3±0.95μm2μm−3) and N. concinna (34.3±4.43μm2μm−3). The surface area of mitochondrial cristae per unit fibre volume was significantly higher in N. concinna, due largely to the greater mitochondrial volume density. These results and previous studies indicate that mitochondrial proliferation in the cold is a common, but not universal response by different species from different thermal habitats. Seasonal temperature decreases on the other hand, leading preferentially to an increase in cristae surface density. Stereological measures also showed that energetic reserves, i.e. lipid droplets and glycogen in the pedal muscle changed greatly with season and species. This was most likely related to gametogenesis and spawnin

Evaluating next-generation sequencing (NGS) methods for routine monitoring of wild bees: metabarcoding, mitogenomics or NGS barcoding

Implementing cost-effective monitoring programs for wild bees remains challenging due to the high costs of sampling and specimen identification. To reduce costs, next-generation sequencing (NGS)-based methods have lately been suggested as alternatives to morphology-based identifications. To provide a comprehensive presentation of the advantages and weaknesses of different NGS-based identification methods, we assessed three of the most promising ones, namely metabarcoding, mitogenomics and NGS barcoding. Using a regular monitoring data set (723 specimens identified using morphology), we found that NGS barcoding performed best for both species presence/absence and abundance data, producing only few false positives (3.4%) and no false negatives. In contrast, the proportion of false positives and false negatives was higher using metabarcoding and mitogenomics. Although strong correlations were found between biomass and read numbers, abundance estimates significantly skewed the communities' composition in these two techniques. NGS barcoding recovered the same ecological patterns as morphology. Ecological conclusions based on metabarcoding and mitogenomics were similar to those based on morphology when using presence/absence data, but different when using abundance data. In terms of workload and cost, we show that metabarcoding and NGS barcoding can compete with morphology, but not mitogenomics which was consistently more expensive. Based on these results, we advocate that NGS barcoding is currently the seemliest NGS method for monitoring of wild bees. Furthermore, this method has the advantage of potentially linking DNA sequences with preserved voucher specimens, which enable morphological re-examination and will thus produce verifiable records which can be fed into faunistic databases

Medical Honey for Wound Care—Still the ‘Latest Resort’?

While the ancient Egyptians and Greeks used honey for wound care, and a broad spectrum of wounds are treated all over the world with natural unprocessed honeys from different sources, Medihoney™ has been one of the first medically certified honeys licensed as a medical product for professional wound care in Europe and Australia. Our experience with medical honey in wound care refers only to this product. In this review, we put our clinical experience into a broader perspective to comment on the use of medical honey in wound care. More prospective randomized studies on a wider range of types of wounds are needed to confirm the safety and efficacy of medical honey in wound care. Nonetheless, the current evidence confirming the antibacterial properties and additional beneficial effects of medical honey on wound healing should encourage other wound care professionals to use CE-certified honey dressings with standardized antibacterial activity, such as Medihoney™ products, as an alternative treatment approach in wounds of different natures