Introducing medical parasitology at the University of Makeni, Sierra Leone

The file attached to this record is the author's final peer reviewed version.Capacity building in Sierra Leone (West Africa) is critical to prevent potential future outbreaks similar to the 2013-16 Ebola outbreak that had devastating effects for the country and its poorly developed healthcare system. De Montfort University (DMU) in the United Kingdom (UK), in collaboration with parasitologists from the Spanish Universities of San Pablo CEU and Miguel Hernández de Elche, is leading a project to build the teaching and research capabilities of medical parasitology at the University of Makeni (UniMak, Sierra Leone). This project has two objectives: a) to introduce and enhance the teaching of medical parasitology, both theoretical and practical; and b) to implement and develop parasitology research related to important emerging human parasites such as Cryptosporidium spp. due to their public health significance. Two UniMak academics, hired to help initiate and implement the research part of the project, shared their culturally sensitive public health expertise to broker parasitology research in communities and perform a comprehensive environmental monitoring study for the detection of different emerging human parasites. The presence of targeted parasites are being studied microscopically using different staining techniques, which in turn have allowed UniMak’s academics to learn these techniques to develop new practicals in parasitology. To train UniMak’s academics and develop both parts of our project, a DMU researcher visited UniMak for two weeks in April 2019 and provided a voluntary short training course in basic parasitology, which is currently not taught in any of their programmes, and was attended by 31 students. These sessions covered basic introduction to medical parasitology and life-cycle, pathogenesis, detection, treatment and prevention of: a) coccidian parasites (Cryptosporidium, Cyclospora and Cystoisospora); b) Giardia intestinalis, Entamoeba and free-living amoebas; c) malaria and d) microsporidia. A theoretical session on common staining techniques was also provided. To facilitate the teaching and learning of these parasites, the novel resource DMU e-Parasitology was used, a package developed by the above participating universities and biomedical scientists from the UK National Health Service (NHS): http://parasitology.dmu.ac.uk/ index.htm. Following the two weeks of training, UniMak’s academics performed different curriculum modifications to the undergraduate programme ‘Public Health: Medical Laboratory Sciences’, which includes the introduction of new practicals in parasitology and changes to enhance the content of medical parasitology that will be subjected to examination. Thus, a new voluntary practical on Kinyoun stain for the detection of coccidian parasites was introduced in the final year module of ‘Medical Bacteriology and Parasitology’; eighteen students in pairs processed faecal samples from pigs provided by the Department of Agriculture and Food Security from a nearby farm. Academics at UniMak used the Kinyoun staining unit (available at http://parasitology.dmu.ac.uk/learn/lab/Kinyoun/story_html5.html; [1]) to deliver this practical. Although our project is at a preliminary stage, it has been shown to be effective in promoting the introduction and establishment of medical parasitology at UniMak and could be viewed as a case-study for other universities in low-income countries to promote the United Nations (UN) Sustainable Development Goals (SDGs) and improve public health understanding of infectious diseases

Applicability of monthly CDC case studies to improve reflection in biomedical science students.

Background Academics from De Montfort University (DMU, UK) in collaboration with other EU universities, virologists and first responders are developing training for health science students to deal with biological incidents. Initially the training is being tested with final year students enrolled on the BSc Biomedical Science (Hons) and in the BMedSci Medical Science (Hons) degree programmes in 2016/17 at DMU. To improve the limited clinical skills of these students, a series of parasitology case studies have been introduced, where students will need to reflect on their knowledge and search for information from different sources to propose possible diagnoses. Reflection is an essential learning tool for developing aspects such autonomous working, critical and analytical thinking and integration of theory with practice (Quintanilla et al., 2016). Methods A series of teaching sessions (theoretical and practical) has been created related to emergency preparedness and response (Peña-Fernández et al., 2016). Students are provided with different scenarios to develop an intervention programme to protect human health in the aftermath of a biological incident involving different biological agents including emerging parasites. During the theoretical component of the training students are provided with different slides collected from the Laboratory Identification of Parasitic Diseases of Public Health Concern (DPDx) (CDC, 2016). Students, by peer group interaction, provide a possible “diagnosis” for each clinical case based on the clinical features presented and microscopic slides provided. Critical thinking is encouraged. Results Although our results are preliminary and we need to evaluate the students’ feedback, the introduction of clinical case-studies has shown to facilitate the acquisition of some clinical skills, particularly in the biomedical science cohort. This is informed by the high level of students’ interaction during these sessions and the formulation of appropriate questions. Students have also shown some gradual improvement in the resolution of clinical case studies throughout the course. Conclusions Despite the fact that student feedback will not be collected until the end of the course, students have display strong engagement and interest in these workshops through interim module level feedback. A priori, these case-studies have been shown to be effective in facilitating the acquisition of different transversal competences including critical thinking, clinical skills, communication and team work

Bayesian Integration of Genetics and Epigenetics Detects Causal Regulatory SNPs Underlying Expression Variability

The standard expression quantitative trait loci (eQTL) detects polymorphisms associated with gene expression without revealing causality. We introduce a coupled Bayesian regression approach—eQTeL, which leverages epigenetic data to estimate regulatory and gene interaction potential, and identifies combination of regulatory single-nucleotide polymorphisms (SNPs) that explain the gene expression variance. On human heart data, eQTeL not only explains a significantly greater proportion of expression variance but also predicts gene expression more accurately than other methods. Based on realistic simulated data, we demonstrate that eQTeL accurately detects causal regulatory SNPs, including those with small effect sizes. Using various functional data, we show that SNPs detected by eQTeL are enriched for allele-specific protein binding and histone modifications, which potentially disrupt binding of core cardiac transcription factors and are spatially proximal to their target. eQTeL SNPs capture a substantial proportion of genetic determinants of expression variance and we estimate that 58% of these SNPs are putatively causal

Fluorescent D-amino-acids reveal bi-cellular cell wall modifications important for Bdellovibrio bacteriovorous predation

Modification of essential bacterial peptidoglycan (PG) containing cell walls can lead to antibiotic resistance, for example β-lactam resistance by L,D-transpeptidase activities. Predatory Bdellovibrio bacteriovorus are naturally antibacterial and combat infections by traversing, modifying and finally destroying walls of Gram-negative prey bacteria, modifying their own PG as they grow inside prey. Historically, these multi-enzymatic processes on two similar PG walls have proved challenging to elucidate. Here, with a PG labelling approach utilizing timed pulses of multiple fluorescent D-amino acids (FDAAs), we illuminate dynamic changes that predator and prey walls go through during the different phases of bacteria:bacteria invasion. We show formation of a reinforced circular port-hole in the prey wall; L,D-transpeptidaseBd mediated D-amino acid modifications strengthening prey PG during Bdellovibrio invasion and a zonal mode of predator-elongation. This process is followed by unconventional, multi-point and synchronous septation of the intracellular Bdellovibrio, accommodating odd- and even-numbered progeny formation by non-binary division

Studying the presence of free-living amoeba in parks and recreational areas in Leicester, UK.

Background: Acanthamoeba spp., Naegleria fowleri, and Balamuthia mandrillaris are considered emerging human pathogenic free-living amoebae (FLA), producing severe infections in both immunocompromised and immunocompetent humans. Although FLA have been found in virtually any environmental compartment, little is known about the potential role of wild animals as reservoirs for FLA. The aims of this study were: a) to identify the presence of FLA in animal faeces collected from urban parks in Leicester (UK); b) to identify if animal samples require pre-concentration to improve their detection. Materials/methods: Duplicates of 50 animal faecal samples were collected in March 2016: 13 from Castle Gardens (LE1 5WH), 28 from Abbey Park (LE4 5AQ) and 9 from the River Soar footpath between both parks. Only fresh samples were selected and collected on days where there had been no precipitation in the preceding 48 h. Results: A veterinarian identified the animal species as 20 avian (10 waterfowl, 8 pigeons, 2 uncertain); 24 canine (15 dog, 8 fox, 1 uncertain), 2 cats, 1 herbivore and 3 unidentifiable. Duplicates of each sample were pre-concentrated using the routine coprological method, modified Telemann and MIF (merthiolate-iodine-formaline). The DNA was extracted from each sample (fresh and pre-concentrated) using the Fast-DNA-Spin kit following previously described methodology. The QIAamp micro DNA extraction kit was used to remove PCR inhibitors. A triplex real-time TaqMan PCR assay was performed to detect FLA; positive controls for the three amoebae were used. All samples assessed for FLA were negative. Conclusions: Results were inconclusive as all samples were negative for FLA. However, understanding the zoonotic potential of animals in the presence and distribution of FLA in the environment is critical to protecting the public from these emerging human pathogens. Although the study of FLA in mammals is rare, studies have described the presence of Acanthamoeba spp. and other FLA in the gastrointestinal tract and faeces of cow, pig, and squirrel representing a potential source of transmission for these opportunistic parasites. Further studies are needed to determine the appropriateness of using pre-concentrating methods in future monitoring studies to determine the presence of FLA in mammals as our results were inconclusive

Building on-line materials for teaching parasitology to health sciences’ students: initial impressions.

Background: It is widely recognised that the use of web-based teaching resources is an increasingly important method for delivering education, and it will be particularly important in the near future due to the progressively increasing number of health science students and the current number of academics in the “European Higher Education Area”. The study of parasitology and infectious diseases is essential to build professionals in the health sector with the key knowledge and skills to face global public health threats such as food-, water- or vector-borne infectious diseases outbreaks. However, the current time dedicated to the teaching of this discipline in all health sciences degrees at De Montfort University (DMU, Leicester, UK) is very little or non-existent depending on the degree/master. Methods: An innovative teaching group at DMU is trying to fill this gap in the currently available teaching offer in line with new trends in global health education, the large number of students enrolled in any health degree and the increasing number of students that would like to study this discipline (but due to different commitments do not have enough time or resources to study on a full time basis). Thus, an innovative teaching group from different EU Universities (DMU and the Spanish universities: University of San Pablo CEU, University of Alcalá, and University Miguel Hernández de Elche) and clinicians (University Hospitals of Leicester, UK) have started to design, create and develop a complete on-line package in Parasitology for undergraduate and postgraduate students that study health sciences. Results: The e-Parasitology package will be accessible through the DMU website (http://parasitology.dmu.ac.uk) in 2017 and will be focused on infection, prevention and treatment of major and emerging parasitological diseases. Conclusions: This teaching resource will aid our undergraduate and postgraduate students to gain a significant knowledge in parasitology by promoting self-learning and internationalization. This poster will explore one of the first mini-modules developed so far related with Toxocara, a helminthiasis with prevalence rates that can reach as high as 40% or more in parts of the world, and the challenges for its development

Exploring the presence of human pathogenic free-living amoebas in different water ecosystems in Leicester, UK.

Background: The presence and distribution of Acanthamoeba spp., Balamuthia mandrillaris and Naegleria fowleri (human pathogenic free-living amoebas, FLA) in different environmental compartments and geographical locations in Europe remains unknown. These FLA can be a public health threat as their cysts are highly resistant to harsh environmental conditions. The aim of this pilot study was to determine the presence of FLA in different water ecosystems close or in Leicester city (Leicestershire, UK) as information on the presence these emerging parasites in the UK is limited in the literature. Materials/methods: A total of 30 water samples were collected from different open water environments in Leicester during winter 2016/17 including: the River Soar and the Grand Union Canal (a canalised section of the River Soar), different lakes highly frequented for fishing or leisure (e.g. John Merricks' Lake, Kings Lears Lake; Bennion Pools Fishing Lake), and a marina near River Soar. The River Soar is rich in wildlife including water birds, fish and plant populations attracting large numbers of users. Water samples were obtained following protocol 1623 described by US EPA and concentrated using IDEXX® Filta Max system following manufacturer's instructions. DNA extraction from concentrated water was performed from each water sample with Fast DNA® Kit. A triplex real-time TaqMan PCR assay was performed to detect FLA; positive controls for the three amoebae were used. Results: All 30 samples assessed for FLA were negative. However these results should be considered as inconclusive as, although rare, several studies have reported the presence of Acanthamoeba spp. in the UK domestic water supplies which may indicate the presence of these human pathogens in other water systems including the environment. Moreover, the incidence of Acanthamoeba keratitis has increased in recent years in England. Conclusions: Further studies will be needed to determine the presence and distribution of FLA in the open water systems monitored to protect the public as recent evidence indicates an increase in infections due to these emerging human pathogens globally. This information is crucial to develop novel strategies to protect humans and increase the awareness of these protozoan parasites in aquatic environments in the UK

Feasibility study of Nb3Al Rutherford cable for high field accelerator magnet application

Feasibility study of Cu stabilized Nb{sub 3}Al strand and Rutherford cable for the application to high field accelerator magnets are being done at Fermilab in collaboration with NIMS. The Nb{sub 3}Al strand, which was developed and manufactured at NIMS in Japan, has a non-copper Jc of about 844 A/mm{sup 2} at 15 Tesla at 4.2 K, a copper content of 50%, and filament size of about 50 microns. Rutherford cables with 27 Nb{sub 3}Al strands of 1.03 mm diameter were fabricated and tested. Quench tests on a short cable were done to study its stability with only its self field, utilizing a high current transformer. A pair of 2 meter long Nb{sub 3}Al cables was tested extensively at CERN at 4.3 and 1.9 K up to 11 Tesla including its self field with a high transport current of 20.2 kA. In the low field test we observed instability near splices and in the central region. This is related to the flux-jump like behavior, because of excessive amount of Nb in the Nb{sub 3}Al strand. There is possibility that the Nb in Nb{sub 3}Al can cause instability below 2 Tesla field regions. We need further investigation on this problem. Above 8 Tesla, we observed quenches near the critical surface at fast ramp rate from 1000 to 3000 A/sec, with quench velocity over 100 m/sec. A small racetrack magnet was made using a 14 m of Rutherford cable and successfully tested up to 21.8 kA, corresponding to 8.7 T

Detection of Ligation Products of DNA Linkers with 5′-OH Ends by Denaturing PAGE Silver Stain

To explore if DNA linkers with 5′-hydroxyl (OH) ends could be joined by commercial T4 and E. coli DNA ligase, these linkers were synthesized by using the solid-phase phosphoramidite method and joined by using commercial T4 and E. coli DNA ligases. The ligation products were detected by using denaturing PAGE silver stain and PCR method. About 0.5–1% of linkers A–B and E–F, and 0.13–0.5% of linkers C–D could be joined by T4 DNA ligases. About 0.25–0.77% of linkers A–B and E–F, and 0.06–0.39% of linkers C–D could be joined by E. coli DNA ligases. A 1-base deletion (-G) and a 5-base deletion (-GGAGC) could be found at the ligation junctions of the linkers. But about 80% of the ligation products purified with a PCR product purification kit did not contain these base deletions, meaning that some linkers had been correctly joined by T4 and E. coli DNA ligases. In addition, about 0.025–0.1% of oligo 11 could be phosphorylated by commercial T4 DNA ligase. The phosphorylation products could be increased when the phosphorylation reaction was extended from 1 hr to 2 hrs. We speculated that perhaps the linkers with 5′-OH ends could be joined by T4 or E. coli DNA ligase in 2 different manners: (i) about 0.025–0.1% of linkers could be phosphorylated by commercial T4 DNA ligase, and then these phosphorylated linkers could be joined to the 3′-OH ends of other linkers; and (ii) the linkers could delete one or more nucleotide(s) at their 5′-ends and thereby generated some 5′-phosphate ends, and then these 5′-phosphate ends could be joined to the 3′-OH ends of other linkers at a low efficiency. Our findings may probably indicate that some DNA nicks with 5′-OH ends can be joined by commercial T4 or E. coli DNA ligase even in the absence of PNK

Creating a model module for the novel resource DMU e-Parasitology.

The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI linkThe study of parasitology has become essential to build future health care professionals with skills to respond to public health threats such as the recent outbreaks due to Cryptosporidium spp. or Giardia in the United Kingdom (UK). To facilitate the teaching of parasitology, which negligible across the different undergraduate and taught masters degrees at De Montfort University (DMU, Leicester, UK), a group from different EU Universities [DMU and the Spanish universities: University of San Pablo CEU (USP-CEU) and University Miguel Hernández], clinicians and practising Biomedical Scientists from the UK National Health Service are developing an on-line package for teaching and learning parasitology named DMU e-Parasitology. This package will have three sections or modules: a theoretical module with mini e-learning units to study major human parasites such as Leishmania spp. or malaria; a virtual laboratory module with units to enhance the learning and study of parasitic diseases such as culture of parasites, staining and use of light microscope to identify these organisms or molecular techniques for the identification of parasites to species level; and a virtual microscope with a real slide collection of clinical samples of parasites. The e-Parasitology resource is being created for undergraduate/postgraduate human health science students, with corresponding degrees of difficulty. Units will include a tool to assess the learning process of the students, in form of a quiz, activity and/or exam, and several formative activities throughout each mini-module. The development of this teaching resource will cover a gap in the traditional teaching and learning methods that are currently used and provided in the participating Universities. The DMU e-Parasitology will aid to our undergraduate students to gain a significant knowledge in parasitology by promoting self-learning. A unit related with Toxocara, a helmintiasis with prevalence rates that can reach as high as 40% or more in different parts of the world, was firstly developed to use as a model for the development of the DMU e-Parasitology. Three undergraduate students that studied parasitology during the first term in 2016/17 [n=27; 6 European Credit Transfer and Accumulation System credits (ECTS); 3rd year module] from the bilingual Pharmacy and Biotechnology degree at USP-CEU were voluntarily recruited to provide comprehensive feedback for this model unit at the beginning of the second term. This unit was tested with these students because of their comprehensive knowledge of parasitology. Students described it as interactive and presenting the appropriate content and resources to study the parasitic disease addressed (toxocariasis). Limitations were the poor navigability in the formative exercise section and the excessive information provided in some slides that could hinder their understanding. The team has addressed these limitations and is using this unit as a model to build the DMU e-Parasitology, which will be accessible through the DMU website (http://parasitology.dmu.ac.uk) in 2018. We consider that this teaching and learning resource will overcome barriers of time, space, equipment and resources; and may help students and scientists around the world in the diagnostic of different parasitic diseases that impact human health