Conservation assessment and monitoring methods for the Annex V Clubmoss group (Lycopodium spp.) in Ireland

Clubmoss species are moss-like plants without flowers. Ireland is home to four members of the Clubmoss group (Lycopodiaceae): Huperzia selago (L.) Bernh. ex Schrank & Mart., Diphasiastrum alpinum (L.) Holub, Lycopodium clavatum L. and Lycopodiella inundata (L.) Holub. All four species are listed as a group in Annex V of the European Union Habitats Directive (92/42/EEC). Annex V lists species whose taking in the wild may be subject to management measures. Under Article 11 of the Directive, each member state is obliged to undertake surveillance of the conservation status of the species in the Annexes and under Article 17, to report to the European Commission every six years on their status. The conservation status of a species is assessed under four parameters: Range, Population, Habitat for the species and Future prospects. This project investigated these four parameters for the four species in the group and devised methods and protocols for the next round of reporting 2012-2018 which is due in 2019.National Parks & Wildlife Servic

Mycolactone Diffuses into the Peripheral Blood of Buruli Ulcer Patients - Implications for Diagnosis and Disease Monitoring.

BACKGROUND: Mycobacterium ulcerans, the causative agent of Buruli ulcer (BU), is unique among human pathogens in its capacity to produce a polyketide-derived macrolide called mycolactone, making this molecule an attractive candidate target for diagnosis and disease monitoring. Whether mycolactone diffuses from ulcerated lesions in clinically accessible samples and is modulated by antibiotic therapy remained to be established. METHODOLOGY/PRINCIPAL FINDING: Peripheral blood and ulcer exudates were sampled from patients at various stages of antibiotic therapy in Ghana and Ivory Coast. Total lipids were extracted from serum, white cell pellets and ulcer exudates with organic solvents. The presence of mycolactone in these extracts was then analyzed by a recently published, field-friendly method using thin layer chromatography and fluorescence detection. This approach did not allow us to detect mycolactone accurately, because of a high background due to co-extracted human lipids. We thus used a previously established approach based on high performance liquid chromatography coupled to mass spectrometry. By this means, we could identify structurally intact mycolactone in ulcer exudates and serum of patients, and evaluate the impact of antibiotic treatment on the concentration of mycolactone. CONCLUSIONS/SIGNIFICANCE: Our study provides the proof of concept that assays based on mycolactone detection in serum and ulcer exudates can form the basis of BU diagnostic tests. However, the identification of mycolactone required a technology that is not compatible with field conditions and point-of-care assays for mycolactone detection remain to be worked out. Notably, we found mycolactone in ulcer exudates harvested at the end of antibiotic therapy, suggesting that the toxin is eliminated by BU patients at a slow rate. Our results also indicated that mycolactone titres in the serum may reflect a positive response to antibiotics, a possibility that it will be interesting to examine further through longitudinal studies

Combined Inflammatory and Metabolic Defects Reflected by Reduced Serum Protein Levels in Patients with Buruli Ulcer Disease

Buruli ulcer is a skin disease caused by Mycobacterium ulcerans that is spreading in tropical countries, with major public health and economic implications in West Africa. Multi-analyte profiling of serum proteins in patients and endemic controls revealed that Buruli ulcer disease down-regulates the circulating levels of a large array of inflammatory mediators, without impacting on the leukocyte composition of peripheral blood. Notably, several proteins contributing to acute phase reaction, lipid metabolism, coagulation and tissue remodelling were also impacted. Their down-regulation was selective and persisted after the elimination of bacteria with antibiotic therapy. It involved proteins with various functions and origins, suggesting that M. ulcerans infection causes global and chronic defects in the host’s protein metabolism. Accordingly, patients had reduced levels of total serum proteins and blood urea, in the absence of signs of malnutrition, or functional failure of liver or kidney. Interestingly, slow healers had deeper metabolic and coagulation defects at the start of antibiotic therapy. In addition to providing novel insight into Buruli ulcer pathogenesis, our study therefore identifies a unique proteomic signature for this disease

Preparation for a first-in-man lentivirus trial in patients with cystic fibrosis

We have recently shown that non-viral gene therapy can stabilise the decline of lung function in patients with cystic fibrosis (CF). However, the effect was modest, and more potent gene transfer agents are still required. Fuson protein (F)/Hemagglutinin/Neuraminidase protein (HN)-pseudotyped lentiviral vectors are more efficient for lung gene transfer than non-viral vectors in preclinical models. In preparation for a first-in-man CF trial using the lentiviral vector, we have undertaken key translational preclinical studies. Regulatory-compliant vectors carrying a range of promoter/enhancer elements were assessed in mice and human air-liquid interface (ALI) cultures to select the lead candidate; cystic fibrosis transmembrane conductance receptor (CFTR) expression and function were assessed in CF models using this lead candidate vector. Toxicity was assessed and 'benchmarked' against the leading non-viral formulation recently used in a Phase IIb clinical trial. Integration site profiles were mapped and transduction efficiency determined to inform clinical trial dose-ranging. The impact of pre-existing and acquired immunity against the vector and vector stability in several clinically relevant delivery devices was assessed. A hybrid promoter hybrid cytosine guanine dinucleotide (CpG)- free CMV enhancer/elongation factor 1 alpha promoter (hCEF) consisting of the elongation factor 1α promoter and the cytomegalovirus enhancer was most efficacious in both murine lungs and human ALI cultures (both at least 2-log orders above background). The efficacy (at least 14% of airway cells transduced), toxicity and integration site profile supports further progression towards clinical trial and pre-existing and acquired immune responses do not interfere with vector efficacy. The lead rSIV.F/HN candidate expresses functional CFTR and the vector retains 90-100% transduction efficiency in clinically relevant delivery devices. The data support the progression of the F/HN-pseudotyped lentiviral vector into a first-in-man CF trial in 2017

The interactions between native bees and alien plants

THESIS 8850This thesis examines the interactions between native bees, in particular bumblebees (Bombus spp.), and the alien plants they visit. Most alien plants are well integrated into the resident plant-pollinator network. They are thus likely to directly and indirectly affect the foraging behaviour, resource use, and the abundance and diversity of individuals, populations and communities of native generalist pollinators as they may augment but also alter the availability and composition of forage resources, by attaining large floral densities and possibly displacing native plants. To date only a limited number of studies have studied the indirect and direct impacts of alien plants on native bees. I investigated the interactions between native bees and alien plants through observational and experimental field studies that took place in sites along the River Liffey and on Howth, Co. Dublin (Eastern Ireland) and in Spiddal, Co. Galway (Western Ireland)

The impacts of an invasive alien plant and its removal on native bees

Although the alien Impatiens glandulifera successfully invades riparian habitats and is visited by native insects, knowledge of its impact on native bees is limited. We assessed pollinator abundance in field sites where I. glandulifera was absent, present or had been experimentally removed. We measured insect visitation to flowers of potted native plants and to I. glandulifera. Bombus spp. comprised the highest proportion of visitors in invaded sites, whereas solitary bees made up the highest proportion in sites where I. glandulifera was removed. More bees, especially medium- and long-tongued Bombus spp. (B. pascuorum and B. hortorum), foraged on I. glandulifera than the native plant species (possibly because the alien was more abundant). We detected no impact of invasion on standardised pollinator abundance, B. pascuorum abundance, nor functional insect diversity, which may be due to variable climatic conditions. We suggest that future studies focus on impacts on rare or specialised pollinator taxa