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<h3><b>2. </b><a href="https://2018.igem.org/Team:Uppsala/Transcriptomics" >Transcriptomics</a> + <a href="https://2018.igem.org/Team:Uppsala/Phage_Display" >Phage Display</a></h3> | <h3><b>2. </b><a href="https://2018.igem.org/Team:Uppsala/Transcriptomics" >Transcriptomics</a> + <a href="https://2018.igem.org/Team:Uppsala/Phage_Display" >Phage Display</a></h3> | ||
<p>As not a lot is known about our worms of interest, we needed an approach that would allow us to detect the worm without knowing its specific markers. For this, we developed new applications of two existing approaches.</p><br><br> | <p>As not a lot is known about our worms of interest, we needed an approach that would allow us to detect the worm without knowing its specific markers. For this, we developed new applications of two existing approaches.</p><br><br> | ||
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<h3><b>3. </b><a href="https://2018.igem.org/Team:Uppsala/Reporter_System" >Reporter System</a></h3> | <h3><b>3. </b><a href="https://2018.igem.org/Team:Uppsala/Reporter_System" >Reporter System</a></h3> | ||
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<h1><a href="https://2018.igem.org/Team:Uppsala/Human_Practices" >The Global Worming Problem and Society</a></h1> | <h1><a href="https://2018.igem.org/Team:Uppsala/Human_Practices" >The Global Worming Problem and Society</a></h1> | ||
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Revision as of 12:40, 16 October 2018
Uppsala iGEM 2018
The Targets
Symptoms
The release of larvae from cysts can lead to lesions, diarrhea, and potential weight loss. This condition is called cyathostominosis [1]. When untreated, the death tolI can reach up to 50%. During the seasonal rupture of cysts, millions of larvae can be released at the same time, which can result in severe and life-threatening consequences [5]. The infection of small strongyles is not one of presence, but one of quantity. They are not dangerous in small amounts and therefore it is difficult to tell whether a horse needs to be treated or not [9].
If farmers had the possibility to know when to treat their horses, prevention of mass rupture and other severe consequences like increased resistance development could be achieved. To reach this goal we have developed a model that (based on multiple parameters) calculates the optimal amount of treatments in a specified period. This will avoid unnecessary use of anthelmintics by raising the awareness in farmers regarding when they actually need to treat their horses.
Current Methods for Detection
Currently the only method for detecting/counting how many worms there are in an animal is counting nematode eggs in fecal samples. This method is not reliable and also requires farmers and ranchers to send in fecal samples to a lab with trained personnel [4]. This technique is expensive, inconsistent, and requires time sensitive shipping of samples. Currently many horse owners are reluctant to conduct the testing; treating their horses regardless of need which contributes to the resistance problem [10].
Resistance Development
Unfortunately, the extensive overuse of deworming drugs has now lead to the detection of worms that are resistant to the most commonly used drugs [2,4]. Since no new deworming drugs have been approved for use in horses the whole equine industry relies on macrocyclic lactones, currently the most common type of deworming drugs used. Unfortunately, cases of resistance among nematode adults have been spotted for macrocyclic lactones as well. So far, four studies with similar results spanning from Europe to North America have been published with concrete data, linking certain small strongyle species to reduced time until detection of eggs after deworming treatment, showing an increase in resistance [2].
Moxidectin is a very common drug used. It is both hazardous for the environment and is also losing its effectiveness towards worm populations [14,15].
Due to all of these facts, we have during our iGEM project developed a reporter that is suitable for the in vivo environment of horse intestines. Moreover, we have developed new applications on existing techniques to be able to find the promoter that would be coupled to the reporter which would at last create the worm buster.
By implementing the model described under Symptoms together with the worm buster, horse owners will not only know how large the treatments should be but also when the treatments should happen. Thus they complement each other to minimize the amount of anthelmintics used; thereby helping to prevent resistance development.
The Worm Buster
The first thing the team did was to define the desired characteristics of the worm buster and outline the project strategy
Requirements
- Our bacteria needs to live in the intestinal tracts of horses.
- They also need the ability to report a quantitative signal of small strongyle in feces.
The small strongyle buster can therefore work as a diagnostic tool and give horse owners the possibility to use individualized dosage of treatment depending on the level of small strongyle infection.
Project Outline
1. Worm Culturing
2. Transcriptomics + Phage Display
As not a lot is known about our worms of interest, we needed an approach that would allow us to detect the worm without knowing its specific markers. For this, we developed new applications of two existing approaches.
-
Transcriptomics
-
Phage Display
4. Modeling
Optimization of the Time Between Treatments
To make sure the unnecessary use of anthelmintics is minimized we have created a model that compares regular and optimized usage of anthelmintics. When referring to the optimized use of anthelmintics, this means that the horse only receives treatment when the amount of parasites exceeds a certain threshold. Our model calculates given the initial amount of worms on the pasture and in the horse. Ideally, the information could be used with the worm buster to get a better overall result of our project. Our solution will not only be able to tell how large the treatments should be but also when they should be administered. This would guarantee the lowest amount of anthelmintics usage, and hopefully help combat the issue of growing resistance. Due to this fact, our model constitutes a large improvement on the potential implementation of our project since it directly brings us closer to our ultimate goal, which is the decrease in resistance development and the improved physical health of horses.
Survey Analysis
When we had chosen to create a worm buster, we wanted to see if there was a market for this kind of product. To gain knowledge about the market we conducted a market analysis together with the human practises group. The modeling group worked with creating a analysis program that could use the information from the market analysis and see if correlations could be found between different variables.
Future Possibilities
Symptoms
When the worms migrate into the arteries they cause inflammation in the arterial wall and induce the formation of blood clots. When a small blood clot forms it may form an embolism which can detach and travel in the bloodstream until it reaches and blocks smaller blood vessels. This prevents oxygen and nutrient supply to surrounding tissues and may result in colic [13]. This can cause problems in the animals, with the worst case scenario being death [3].
Having these parasites is obviously a huge problem and there is great need for methods that can detect the parasites before ingestion, preventing infection. This makes the need for the second worm buster even larger.
Current Methods for Detection
To be able to count either small or large strongyles, the current methods require growing their eggs from samples and counting them. The disadvantage is that this method only works for adult strongyles that have already laid eggs! This technique only makes it possible to detect strongyles after the infection which is a huge disadvantage! Other methods for detection, such as using an ELISA test or a PCR based method are expensive and require a high level of expertise [11, 12, 16].
In conclusion, the current need is a simple detection method which could be applied to grass samples to be able to check if a pasture is secure for horses to graze on. Therefore we have analysed the possibility of developing the second worm buster by studying fluorescence in grass samples. Our results were promising since the fluorescence was clearly visible in the grass!
Raising Awareness
We have attacked the problem at several angles to raise awareness for our project. We have performed a market analysis of the problem and sent out questionnaires to better judge the severity of the problem. We’ve even been published in a local swedish magazine, explaining the problem and raising awareness locally.
Financial perspective
The equine industry includes many areas like stud farms, breeding, slaughter, trading, training and sports stables, and riding schools [18]. In Sweden the equine industry has a turnover of 450-500 million EUR and horses graze around in fields that in total correspond to 600,000 football fields [17]. There are approximately 350 000 horses is Sweden (2016) which gives Sweden a larger horse/person ratio than many other European countries [18, 19]. A large amount of money is spent on horses within and without the borders of the EU, so this is clearly a global problem.
Even though all the available data points at a high demand for our product, more data is needed to predict how an eventual new GMO product would be perceived. That is why we have worked on the market analysis and economics modelling. The result from these decides if the eventual solutions actually will be possible to implement in real life.
Uppsala 2018 is on a mission for the future of iGEM
In iGEM Uppsala we are worried about the future of iGEM members wellbeing and have seen multiple cases of burnouts. This is something that has to be changed to make iGEM sustainable for the future. To combat this problem we have hosted an event with iGEM Stockholm and Lund which shed light on the growing problem of burnouts and how to prevent them. To further spread the knowledge we have published a booklet about it! The booklet is a summary of useful guidelines and techniques to prevent team members from burn outs. It also speaks about the current situation in iGEM which we have summarised after performing an international survey with answers from current team members and previous team members all over the world.
References
[1] Karlsson J. Parasite detection in extensively hold Gotland ponies. 50. Source: https://stud.epsilon.slu.se/7939/7/karlsson_j_150622.pdf
[2] Molena RA, Peachey LE, Di Cesare A, Traversa D, Cantacessi C. 2018. Cyathostomine egg reappearance period following ivermectin treatment in a cohort of UK Thoroughbreds. Parasites & Vectors 11: 61. Source: https://parasitesandvectors.biomedcentral.com/articles/10.1186/s13071-018-2638-6
[3] Andersson E. Hur påverkas prevalensen av selektiv avmaskning? 22.
[4] Traversa D, von Samson-Himmelstjerna G, Demeler J, Milillo P, Schürmann S, Barnes H, Otranto D, Perrucci S, di Regalbono AF, Beraldo P, Boeckh A, Cobb R. 2009. Anthelmintic resistance in cyathostomin populations from horse yards in Italy, United Kingdom and Germany. Parasites & Vectors 2: S2. Source :https://parasitesandvectors.biomedcentral.com/articles/10.1186/1756-3305-2-S2-S2
[5] Matthews JB, Hodgkinson JE, Dowdall SMJ, Proudman CJ. 2004. Recent developments in research into the Cyathostominae and Anoplocephala perfoliata. Veterinary Research 35: 371–381. Source: https://www.ncbi.nlm.nih.gov/pubmed/15236671
[6] Jasovský D, Littmann J, Zorzet A, Cars O. 2016. Antimicrobial resistance—a threat to the world’s sustainable development. Upsala Journal of Medical Sciences 121: 159–164.
[7] K. Nielsen M, Andersson U, K. Howe D. 2015. Diagnosis of Strongylus Vulgaris. University of Kentucky
[8] C. Sellon D, T. Long M. Equine Infectious Diseases. Elsevier Health Sciences, 2007 Source: https://books.google.se/books?id=sZku4lppRfsC&pg=PA483&lpg=PA483&dq=Cyathostominae+horse&source=bl&ots=GtJJjXA5Kd&sig=9ktoAaiQnVSd6xTC_hhnvujvUBE&hl=sv&sa=X#v=onepage&q=Cyathostominae%20horse&f=false
[9] Colen MA, D. C. K van D, F. N. J. K. Anthelmintic resistance in Cyathostominae. Source: https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=20&ved=0ahUKEwj-xuHwxuDbAhXGFZoKHUsSBmM4ChAWCGQwCQ&url=https%3A%2F%2Fdspace.library.uu.nl%2Fbitstream%2Fhandle%2F1874%2F289404%2FOnderzoeksstage%2520anthelmintic%2520resistance%2520in%2520Cyathostominae%2520MAColen.doc%3Fsequence%3D2%26isAllowed%3Dy&usg=AOvVaw2PhKtv4XqGJTqJg86aFIFd
[10] Johnstone DC. 2000. S. vulgaris pathogenesis. WWW-dokument 2000-: http://cal.vet.upenn.edu/projects/merial/Strongls/strong_8d.htm. Retrieved 2018-06-20.
[11] Ling J. 2017. Strongylus vulgaris och Anoplocephela perfoliata. WWW-dokument 2017-07-14: https://stud.epsilon.slu.se/10486/.Retrieved 2018-06-20.
[12] Bracken MK, Wøhlk CBM, Petersen SL, Nielsen MK. 2012. Evaluation of conventional PCR for detection of Strongylus vulgaris on horse farms. Veterinary Parasitology 184: 387–391.
[13] 2013. New Method for Detecting Bloodworms. WWW-dokument 2013-07-27: https://thehorse.com/116401/new-method-for-detecting-bloodworms/. Retrieved 2018-06-21.
[14] Cobb R, Boeckh A. 2009. Moxidectin: a review of chemistry, pharmacokinetics and use in horses. Parasites & Vectors 2: S5.
[15] Corning S. 2009. Equine cyathostomins: a review of biology, clinical significance and therapy. Parasites & Vectors 2: S1.
[16] Nielsen MK, Scare J, Gravatte HS, Bellaw JL, Prado JC, Reinemeyer CR. 2015. Changes in Serum Strongylus Vulgaris-Specific Antibody Concentrations in Response to Anthelmintic Treatment of Experimentally Infected Foals. Frontiers in Veterinary Science, doi 10.3389/fvets.2015.00017.
[17] The Swedish horse sector. WWW-dokument: https://hastnaringen.se/swedish-horse-sector/. Retrieved: 2018-06-23.
[18] Häggblom M, Rantamäki-Lahtinen L, Vihinen H. Equine sector comparison between the Netherlands, Sweden and Finland. 36. Source: http://www.hippolis.fi/UserFiles/hippolis/File/EquineLife/equine_sector_comparison_between_the_netherlands_sweden_and_finland.pdf
[19] Hästar och anläggningar med häst 2016 - JO24SM1701 - In English. WWW-dokument: http://www.jordbruksverket.se/webdav/files/SJV/Amnesomraden/Statistik,%20fakta/Husdjur/JO24/JO24SM1701/JO24SM1701_inEnglish.htm. Retrieved: 2018-06-24.