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Introductie
One Health PACT / Avans – Lab-students

Image of a mosquito on a plant

13 3rd year Biology and Laboratorial Research students are till the end of January working on OHPACT-related projects within Avans under the supervision of teachers with One Health PACT as their client in the minor bio-informatics. Within this part of the curriculum of the bachelor education, project based learning is essential. In collaboration with Erasmus MC (Bas Oudemunnink and Reina Sikkema), students are contributing to the species identification of ticks in the Netherlands. In association with Radboud (Tom van der Most), students are developing data-analysis pipeline to determine the presence of RNA viruses in mosquito samples.

Who are these students?

Roy van Engelen

Who am I? 
My name is Roy van Engelen. I am 22 years old bachelor student at the university of applied sciences Avans hogeschool, where I am currently in the third year specializing in patient oriented research.  

What subject will I be focussing on this semester? What will I be researching? 
The subject that I will be focusing on this semester is to determine the detecionlimit of mosquitoes in heteropteras with DNA extraction and real- time qPCR.

What do I want to do with this in the future? 
To be honest only the techniques and the way how projectsetups work from A to Z, because after this degree I want to do a master or study medicine at a university, where I want to specialize in different sectors.

What would I like to learn?

I would like to learn to start a project from the ground up and work on it as much as possible to learn a lot about time management and responsibility of working without help.


Demi Struijk

Who am I? 
My name is Demi Struijk, a nineteen-year-old student at Avans University of Applied Sciences. I am now in my third year of studying biology and medical laboratory research. Currently, I specialize in patient-oriented research.

What subject will I be focussing on this semester? What will I be researching? 
I am currently involved in a project investigating the relationship between predators and mosquitoes. We focus on determining the detection limit of the iDNA from mosquitoes in true bugs using real-time PCR. In this way, we can examine whether the predators can be used as a possible natural pesticide against mosquitoes.

What do I want to do with this in the future? 
I’m not quite sure what I want to do in the future yet. I am certainly interested in patient-oriented research. I want to do something with this later on, but there are many different sectors and techniques, which makes it difficult to choose for now.

What would I like to learn?
As I said, my interest lies more in patient-orientated research. But I’d like to learn about independently setting up surveys, which we’ll be working on a lot this semester


Laura van Schouwenburg

Who am I? 
I am Laura van Schouwenburg, 22 years old and I am currently specializing in patient-oriented research in the third year of the study biology and medical laboratory research at Avans University in Breda.

What subject will I be focussing on this semester? What will I be researching? 
I am currently researching the effect of infection with the Usutu virus on the gene expression of interleukin 1 beta in blackbirds together with Sophie Wouters. This research is being done in collaboration with the University of Utrecht. During this study, we work with real-time PCR and use histological staining to investigate necrosis in the brain.

What do I want to do with this in the future? 
I am still orientating myself within this field, but I already know that my interests lie in researching patient material.

What would I like to learn?

During this project we are mainly engaged in investigating the difference in gene expression between blackbirds that have been infected and blackbirds that have not been infected. Therefore, my main focus is on learning how to investigate this using real-time PCR and associated calculations.

You can find me on LinkedIn


Cheyenne van Kraaij

Biology and medical (laboratory) research

Who am I? 
Hello, my name is Cheyenne van Kraaij and I am 26 years old. I am studying Biology and medical laboratory research at Avans University Breda. I am doing the specialisation patient oriented research on this moment.

What subject will I be focussing on this semester? What will I be researching? 
The research is aimed to achieve a rapid detection method of arboviruses in mosquitoes. Currently, several PCR protocols have been developed that are able to detect virus RNA in birds and mosquitoes, but these protocols are too time consuming to implement on a large scale. In addition, this methods requires gel electrophoresis equipment, which is not easy to deploy in the field. Also, the anatomical effects that the viruses have on the host (mosquito) are not fully known and for this reason additional research is being conducted into the anatomy of the mosquito.

The aim of this project is to test whether a DNA/RNA quantification using the Qubit allows rapid identification of positive PCR reactions with reduced gel electrophoresis usage. This is done by isolating the obtained mosquito virus RNA by the TriPure method, then the concentration of the isolated RNA is retrieved with the Tapestation, after the Tapestation the RNA concentration and purity is determined with the Qubit RNA BR detection. The RNA is then converted to cDNA with Reverse Transcirptase (RT) to perform an RT-Endpoint-PCR. These PCR products are analyzed with the Qubit dsDNA HS. With the results of the QUbit, a detection limit can be set and the so-called cutoff value can be determined for the arbovirus research. After this, the virus cDNA will be examined with the Nanopore sequencing in order to identify and characterize the samples. The arboviruses included in this study are West Nile, Usutu and Sincebis. As additional research for this project, there will be an investigation to examine if it’s possible to make the mosquitoes visible underneath the microscope by the paraffin method, in particular, the focus will be on the brain.

What do I want to do with this in the future? 
I like to develop myself in various research methods to find out wich research field suits me the best. For this reason I like to try many different research area’s, especially the ones I am not familiar with on this moment. 

What would I like to learn?

Like I told in the last question I would like to learn various new methods and research subjects so I can found out wich research area fits me the most. So I like to aim on methods that are still unknown for me. So I like to get known with the unknown.

You can find me on LinkedIn


Samuel Odro

Who am I?
I am Samuel Odro, 22 years old and living in Tilburg. I study Biology and medical lab research at Avans University, I’m in my 3rd year. I love Histology, so that’s something I will be focusing on this study. In my free time I like to do weightlifting, reading and gaming.

What subject will I be focusing on this semester? What will I be researching?
arboviruses and the surveillance of them. RNA isolation with a TRI pure kit, RT-End point PCR, QUBIT PCR (for viral ssRNA and cDNA) and gel electrophoresis will be performed. This to see if the QUBIT can deliver a quick and reliable PCR result without the need of gel electrophoresis. Part of this research is determining the cut-off value for a positive PCR result. This is done with arthropods from Erasmus MC infected with these 3 arboviruses; West-Nile virus, Usutu virus and Sindbis virus. 

Also the effect of these arboviruses on the anatomy (especially the brain) of arthropods will be
researched histologically. Arthropods from Wageningen University will be processed and stained with the; H&E and Alcian blue-PAS staining. The presence of malaria plasmodium will be researched by a May-Grunwald Giemsa stain. 

What do I want to do with this in the future?
I would like to apply any techniques that relate to histology or laboratory animal science

What would I like to learn?
Virology, the working mechanisms of viruses

You can find me on LinkedIn


Alisha Biharie 

Who am I?
My name is Alisha Biharie and I am 21 years old. I am in my third year of biomedical sciences at Avans University of applied sciences in Breda, and I am currently following a specialization in patient-oriented research. After the specialization I intend to follow a minor about immunology and infectious diseases at Leiden University of applied sciences. 

What subject will I be focusing on this semester? What will I be researching?
During this semester I will focus on the effect of the Usutu virus on the gene expression of interferon gamma in blackbirds. In collaboration with Utrecht University and the OH-pact, this research can be performed. During this project the endpoint PCR, RT-PCR and real-time PCR techniques will be used to  analyse the gene-expression. The potential damage to the brain tissue caused by the Usutu virus will be examined by performing a H&E staining. 

What do I want to do with this in the future?
After this project I would like to continue studying infectious diseases and their effect on the host on both micro and macro level. 

What would I like to learn?
I would like to learn more about the mechanism of the Usutu virus and other viruses responsible for infectious diseases. I wish to know more about the ability of the viruses to infiltrate host cells and get past the immune system.

You can find me on LinkedIn


Fleur Nuijten

Who am I?                                                                                                                           
I’m Fleur Nuijten, a 19-year student in my third study year of forensic research at Avans University in Breda. Currently I’m following a minor about patient centred research. My plan is to do a specialization in forensic biology in the second half of this year. 

What subject will I be focussing on this semester? What will I be researching?
This semester I will be preforming a project concerning the Usutu virus in blackbirds. The gene-expression of toll like receptor 3 (TLR3) in healthy and USUV infected birds will be analysed with a qPCR. The resulting data is compared with each other to determine a possible change in TLR3 activity after USUV infections. 

What do I want to do with this in the future? 
In the course of this study, I have discovered multiple subject that have gained my interest. These are mainly subjects in the biological sector of my study like: microbiology, histology and pathology. I’m still uncertain on what exactly I would like to do in the future, but I hope this project can help me orientate myself about other subjects and disciplines that might suit my interests.

What would I like to learn?

There’s a couple of new methods that will be performed during this project, mainly concerning real-time PCR and the analysis of gene expression. I think this project will teach me more about gene-expression, how to research it, and how it works. Furthermore, I hope to learn more about viruses and infections and how to treat them. 

You can find me on LinkedIn


Hanae

Who am I? 
My name is Hanae, and I am 21 years old. I am a third-year biomedical research student at Avans Hogeschool. 

What subject will I be focussing on this semester? What will I be researching? 
I am currently working on a project with Utrecht University for my specialization in Patient-oriented research. The project aims to detect a potential difference in interferon- γ gene expression in infected blackbirds with the USUTU virus and non-infected blackbirds using molecular diagnostics.

What do I want to do with this in the future? 
My interest definitely lies in public health, and I would love to help people around the world.  

What would I like to learn?
Due to global warming, the spread of exotic diseases is growing. To make a change and leave a beautiful planet for the children of tomorrow, it is important for me to gain more knowledge about this topic. I also want to learn more about the role of mosquitos in transferring the viruses and the different immune pathways that are involved during USUV infection in birds.

You can find me on LinkedIn


Floor Steinvoort 

Who am I? 
My name is Floor Steinvoort and I’m twenty years old. I am in the third year of my biomedical research bachelor at the university of Avans Breda. I recently chose to specialize in patient-orientated research as of this year. After this I’d like to continue my studies by doing a minor targeted on animal testing and maybe do an internship abroad. 

What subject will I be focussing on this semester? What will I be researching? 
Because of the specialization I chose I’m doing a project targeting arboviruses, specifically ones spread by mosquitoes. This project was commissioned by the Erasmus Medical Centre’s department of Viroscience. Our goal is to develop a protocol for the quick detection of the arboviruses, Usutu, West-Nile and Sindbis using a Qubit. This kind of protocol could hopefully eliminate most of the need for other slower and more labour intenstive methods. 

What do I want to do with this in the future? 
I’m not sure what the future will hold for me as I am still trying the figure out what I want to do. What I am sure of is that I want to work in the patient orientated field, even if I haven’t figured out the kind of research yet.

What would I like to learn?
I’d like to learn more about what working in viroscience is like, since I have never done a project surrounding the topic before. Also I’d like to explore the diffrient protocols and methods that we are able to use. As we get free reign it would be a good opportunity to not only learn about new protocols but also learn about what setting research up from scratch is really like. 

You can find me on LinkedIn


Jip van Gurp

Who am I? 
My name is Jip van Gurp. I’m a 20 years old student in my third year of biology and medical laboratory research at Avans University in Breda. Currently I’m specializing in patient-oriented research. 

What subject will I be focussing on this semester? What will I be researching? 
The subject I’m focussing on is about the Usutu virus in blackbirds. The gene-expression of toll-like receptor 3 (TLR3) in infected and healthy blackbirds will be analyzed through a qPCR. With this information, it can be determined whether USUV is changing the  expression of TLR3 genes or not.  

What do I want to do with this in the future? 
I don’t have a specific plan for the future. I think the biology pathway is the pathway that I’m most interested in. think of histology or pathology. And specific the patient-oriented research, like what I’m specializing in now. 

What would I like to learn?
Gene-expression does interest me a lot. I would like to learn more about the gene-expression in different organisms and how different infections may cause a change in the expression of these genes.

You can find me on LinkedIn


Mike van Griethuijsen 

Who am I? 
I am Mike van Griethuijsen and I am 20 years old. I am currently in my third year at Avans Academy of life sciences and technology studying to become a Bachelor of biomedical research. After this I’d further expand my studies by following a pre-master and eventually a Master in biomedical research and technology.

What subject will I be focussing on this semester? What will I be researching? 
I am currently invested in a project where the aim is to use realtime PCR to determine if eDNA analysis could be used to find mosquito infested waters. Knowing this could play a vital role in fighting mosquitoes and the arboviruses they help transfer.

What do I want to do with this in the future? 
I am not yet certain what the future will hold for me. However I doubt it will contain eDNA analysis. My current dreamjob would probably be overseer for a diagnostic/medical laboratory

What would I like to learn?
I would like to learn more about realtime PCR and eDNA analysis aswell. This project has succeeded in grabbing my interest when I was not that excited the first couple of weeks. Since this project alongside OH pact could drastically change the world for the better I am happy to be a part of it.


Naomi Huisman 

Who am I? 
My name is Naomi Huisman and I am 19 years old. I study biology and medical laboratory research majoring in forensic laboratory research at Avans University of Applied Sciences and I am in my third year. Currently, I am doing a minor in patient-centered research.

What subject will I be focussing on this semester? What will I be researching? 
This semester I will be working on a project optimizing the use of the Qubit in detection of arboviruses. The goal is to design a suitable protocol where the Qubit works optimally for detecting Sindbis, Usutu and West-Nile viruses. Surveillance of the spread of these viruses by mosquitoes requires the examination of large numbers of mosquitoes. Hopefully, other slower and more lab-intensive methods will no longer be needed when an optimized protocol of the Qubit is designed. 

What do I want to do with this in the future? 
Working with issues within virology is entirely new to me. Through this project, that was commissioned by the Erasmus Medical Centre’s department of Viroscience, I see the importance of researching fast and efficient methods for the surveillance. However, I notice in that I am more interested in diagnostics and not so much in researching better protocols and method. I hope to work in health care later, perhaps in virology, who knows. 

What would I like to learn?
As I mentioned, I have no experience in virology yet. By working on this research, I hope to gain more knowledge of virology. I also hope to learn a lot from optimizing. How do I create a good setup to test out an instrument like the Qubit and then how do I make it work optimally? By thinking about this carefully, I hope to learn a lot in the research field.

You can find me on LinkedIn


Sophie Wouters

Who am I? 
I am Sophie Wouters, 19 years old and I am studying biology and medical laboratory reasearch, with patient-oriented research as specialization at Avans university in Breda. And I am in my third year now.

What subject will I be focussing on this semester? What will I be researching? 
the subject I am focussing on this semester is the Usutu virus in blackbirds. And the gene expression of interleukin 1 beta in infected blackbirds and healty blackbirds. The thechniques we will use are real time PCR and we will do histological stainings on the brain of the blackbirds.

What do I want to do with this in the future? 
I am very interested in neurobiology and histological stainings and hope to do something with that in the future.

What would I like to learn?
I am not very familiar with gene expression projects, but I would like to learn more about the change in gene expression of interleukin 1 beta between infected blackbirds and healthy blackbirds.

You can find me on LinkedIn


Joris Goddrie

Who am I? 
I am Joris Goddrie I’m 21 years old. I am currently studying at avans, I’m doing my specialisation in patient-centered research. After this achlor I would like to further expand my knowledge by doing a master in Biomedical research.

What subject will I be focussing on this semester? What will I be researching? 
In my current project I am trying to research if I can determen if I can see how menny musquitos live in a water source and where they procreate using eDNA and realtime-PCR.

What do I want to do with this in the future? 
I think realtime-PCR cou;d play a bbig role in my future because I think it wil be a big part of the detaction of a lort of deceases and it wil be essencial in the research to fight desices.

What would I like to learn?
I would like to learn to wordt with the realtime-PCR and how it can be implemented for the best results further I would like to use my skills and knowlige to help the OH pact fight mosquitos and arboviruses.