Research Proposal: Investigating links between Primary Rainforest Loss and the Zoonotic Disease Ebolavirus.

Introduction & Rationale of project:

This study investigates the link between loss of primary rainforest habitat in Liberia and the increased risk of zoonotic diseases.
We are targeting 3 bat species, as suspected disease reservoirs leading to human transmission of Ebolavirus (1) and will be comparing their infection rate in two locations: a primary rainforest in a protected nature reserve vs. a severely degraded region with significant human impacts. ‘Primary’ refers to forest that exists in original condition, unaffected by human activities.
By communicating findings to the Liberian Government, Forestry Development Authority (FDA), The Forest Trust (TFT), and regulatory bodies managing deforestation, we will exhibit demonstrable cost to human health of deforestation.

The specific objectives are:

  • Determine whether correlations exist between loss of Primary rainforest and infection rate of Ebolavirus in the bat species: Hypsignathus montrosus (Hammer-headed fruit bat), Epomops franqueti (Franquet’s epauletted fruit bat), Myoncteris torquata (Little collared bat);by measuring the infection rates in these bat species in an area of high deforestation and habitat degradation, comparatively with an area of primary rainforest in a protected nature reserve.
  • Once the hypothesis has either been confirmed or nullified, we will examine the potential repercussions of our findings, possibly presenting the case that reducing deforestation could lead to fewer human breakouts of Ebola.
  • Before we could present the case that deforestation creates a greater risk of human Ebola outbreaks, a follow-up study would be needed to connect the level of bat infection, and level of human infection.

Our results will be cooperatively written by team members, then peer reviewed. If successful, our aim is to present these findings to the regulatory bodies responsible for controlling deforestation in Liberia and demonstrate the clear risks to human health of excessive primary forest loss. 

This study will make significant contributions to our understanding of zoonotic disease spread in tropical regions.
There is moral imperative to investigate the changes our species is causing to the geographical landscape of biologically significant areas such as rainforests, and the subsequent consequences.

This significance is heightened when findings may be at the detriment to the health of thousands of people. There have been 26 recorded Ebola outbreaks between 1976-2014 with a mortality rate of 39-88%, fluctuating over time and geographically (Fig.1).

The rationale behind the creation of this study follows a report by Olivero (1), which hypothesised that due to loss of habitat and resources linked to deforestation, animals previously able to widely disperse and live in separate colonies (in the complex habitat that is primary rainforest), are forced to coexist and directly compete more frequently, increasing likelihood of disease transmission.

This hypothesis is reinforced because Ebola is transmitted through contact with infected bodily fluids, overcrowding and increased interaction (such as in cities), has historically led to higher infection rates (2). This has been proven in animals: infection and deaths in great apes rise during the dry season, when water and fruit is scarce, forcing interaction between groups or individuals that may have never come into contact before, competing for resources (3).  

If we sum these rationale,
– increased contact leads to higher transmission,
– the destruction of primary rainforest causes increased contact between animals,
– there is high deforestation in Liberia (Fig.2),
– Liberia was one of the epicentres of the last Ebola epidemic,
– the 3 bat species we have identified are confirmed carriers of this disease,
we can understand the hypothesis that deforestation could correlate with increased outbreaks of Ebolavirus.

When examining the statistics of deforestation in Liberia, we see it’s rate accelerating. Between 2001-2010, 5,958 hectares of primary forest was lost, and between 2011-2018, 18,837 hectares was lost. These figures show the exponential nature of primary forest loss in the region, in the last 7 years, 12,879 hectares more was destroyed than the previous 9 years. Overall, 14% of Liberia’s primary rainforest has been lost (4). As the three species being studied as vectors are tree-roosting bats, this will cause a severe reduction in habitat.


This is a preliminary study, with the long-term goal to conclude whether high deforestation ultimately causes increased Ebolavirus outbreaks in humans. A subsequent investigation on the relationship between bat infection rates and human infection is needed.

Higher death rates could provide proof for companies with interests in deforestation, that their actions are causing tangible damaging effects, for flora, fauna, and humans in the region.

Furthermore, with the current state of affairs around the COVID-19 pandemic, there is focus on the scientific community to understand the occurrence and transmission of zoonotic diseases, in order to prevent events such as this. I believe a case could be made that this proposal begins the first steps of research into long-term strategies to avoid the excessive spread of zoonoses, namely the protection of primary rainforests.

Basic Methodology:

  1. Set up base camp and field laboratory in Lofa-Mano National Park, in prime location for all 3 bat species. They’re prevalent in this area, as you can see from the map of their geographical distribution (Fig. 3)
  2. Set up 3 mist nets, in clear flight paths through the trees.
    We aim to create randomly selected sample groups from the species at each location, to represent the infection rate of the whole population. We will remove sampling bias by placing the nets in areas heavily frequented by all species, such as routes near nesting sites; this way we can ensure that the animals we catch aren’t just the weaker individuals who have strayed into our traps, so the sample groups are an accurate representation of the population.
  3. Each morning pull down the mist nets. Identify whether any individuals of the three target species are caught in the nets. Transport relevant animals to the field lab, release any by-catch.
  4. Humanely euthanise the animals, using anaesthesia and cardiac puncture. Requirements include anaesthetic agent, towel, cotton, 19-25G needle with 1-5 ml syringe, surgical blade, tube (internal diameter of 0.1 to 0.3 mm) for thoracotomy, plastic disposable bag and blood sample collection tubes. During blood sample collection, animal will be in terminal anaesthesia. Appropriate needle is used for blood sample collection with or without thoracotomy. Blood sample will be taken from the heart, preferably from the ventricle slowly to avoid collapsing of heart.
    We must euthanise the animals because in order to run virus isolation, we must have blood serum, and in order to create a serum we need a high volume of blood sample, which in itself would severely damage the bat. In addition, the locations with the highest viral load in a body infected with ebolavirus are the liver and spleen. We need these tissues to run the DNA amplification analysis later.
  5. Work throughout the day to preserve the relevant tissues from every animal caught that night. Serum will be obtained by centrifugation of whole blood and stored in liquid nitrogen for transportation. Necropsy specimens of liver and spleen from each animal is placed in Cryovials and immediately frozen in dry nitrogen.
  6. Repeat procedures 2-5 every day for our time in this location.
  7. Pack up camp, field lab, and specimens. Travel to Totota & set up base camp and field lab.
  8. Repeat steps 2-5 for five days.
  9. Travel to The University of Liberia, in Monrovia.
  10. Once at the university, begin analysis of specimens using their specialist equipment.
  11. Serology, Virus Isolation, Immunohistochemical Staining, and DNA Amplification and Sequencing will be carried out at the facilities provided in the department of ‘Veterinary Public Health’.
    These tests will be carried out in cooperation between our team, Professor Olayinka Ishola and Professor Sampson Chea; we will be led by the Professors at the Medical School, as well as their students. We will follow the basic methodology from the Leroy 2005 paper to carry out our specimen analysis, as this paper produced successful results (3) (The Leroy methodology can be found in the Supplementary Information segment at the end of this document).

Once back in the UK, we will do statistical analysis to determine whether there is a higher proportion of Ebola-infected bats in location A (Lofa-Mano National Park – area of pristine primary rainforest) or location B (Totota – area of highly degraded rainforest).
If there is a significant difference in this proportion, we can make conclusions as to what this means, if not, the null hypothesis will be accepted, saying there is no correlation between rainforest quality and Ebola infection in these 3 virus-reservoirs.

Project Advisors & in-country Associates

Dr Mika Peck – Senior Lecturer in Biology, University of Sussex.
Mika provided guidance with the development of my project, such as methodologies and logistics, as well as providing specialist information on conservation programmes in tropical rainforests. Some relevant studies Mika has spearheaded are The Santa Lucia Reserve in the Ecuadorian Andes, the Choco Project in Ecuador, and involvement in indigenous communities in Papua New Guinea.

Professor Olayinka Ishola – Lecturer in Biology, Head of Department of Veterinary Public Heath, University of Liberia Medical School.
Professor Ishola is a specialist in epidemiology, virology, and veterinary bacteriology. She is our lead associate in-country, due to her extremely relevant skill set. Additionally, Professor Ishola is the lead of a Lab with all the relevant equipment that we will need to carry out the analysis of our collected specimens.

Professor Sampson Chea – Lecturer in Biochemistry and Molecular Biology – University of Liberia.
Sampson has provided guidance for the analytical methods of our specimens, due to his detailed knowledge of molecular biology in the context of disease. He will assist and oversee the project stage at the university.

Sayamajunkon S.E Forkay – BSc Biology, University of Liberia.
Published a significant paper in 2019: “Serological evidence of Ebola virus exposure in dogs from affected communities in Liberia: A preliminary report”. Sayamajunkon is a student in Professor Ishola’s lab, and she has successfully carried out and published a report on the virus isolation of Ebola in a serum form, from animals. Due to these connections, it will be very useful to have her assistance at this stage of the methodology. Additionally, Sayamajunkon’s successfully published methodology has provided guidance to our study.

University of Liberia – Medical School (Achille Mario Dogliotti College of Medicine)
Our partnership with the university is integral to this investigation. The staff and students here are providing valuable guidance and assistance to the Lab portion of the methodology, as well as lending us essential scientific equipment to carry out the data/sample collections in the field. Collaborating with the university and its associates is essential to the running of this study.

Lofa-Mano National Park
One of the key national parks of Liberia, the other being Sapo National Park. The zone was protected in 1979 and was declared the most species abundant area of the country. The site is 890m2, and is home to thousands of species, with the aim of maintaining an area of pristine, unexploited, primary rainforest.
This park will be key to our investigation, and we are grateful to the rangers and guardians for allowing us permission to work here.

Green Advocates of Liberia
This is a charitable organisation based in Liberia, working to promote ecological awareness, in combination with human rights issues. They have connections to many parallel organisations and works with a wide range of national and international civil society organizations to promote public disclosure of financial transactions in the extractive industries and forestry sectors. Most significantly they were heavily involved in drafting the 2006 National Reform Forestry Law, which enabled management and conservation of forest resources of Liberia, regulates commercial and other use of forests resources, provides for the protection of the environment and wildlife in forests, regulates the trade in forest products and provides for various other matters relative to forestry and wildlife. We will work with the Green Advocates to disseminate our findings to relevant bodies.

The Forest Trust
TFT is a global environmental conservation charity, with offices in Liberia. This organisation is deeply concerned with the effects of the palm oil industry, and its effects on ecosystems and human communities. The TFT has connections to the point of raw material extraction (forest, plantation, quarry), to ensure products are traceable throughout the supply chain and that people and environment are respected. They also work with key business leaders and supply chain partners, to lobby and learn how to convince these people to source products more responsibly. The collaboration with TFT in the latter stages of this investigation, producing an article for their newsletter that summarises our findings and their consequences, will help us reach a wider and more powerful audience.

Bats without Borders
This organisation works in Africa to gather information on the whole Chiropteranfamily, with specific focus on ecology, conservation, human impacts, bioindicators, and bat disease. They have a very wide reach in terms of public engagement and education, so will be incredibly useful in terms of data dissemination and have shown interest in the results of our study. The endorsement from this body adds weight to our findings and methodology.

Team Members:

NameNationalityAffiliation/ OrganisationQualifications, Skills, Experience relevant to the methodology.Role in Team
Miss Amy HowardBritishUniversity of Sussex.
BSc Zoology.
Bachelor’s degree in Zoology, specialisation in disease ecology.
Proficient with SPSS.

Team Leader
Miss Ruby ArthurBritishUniversity of Sussex. BSc Biology.Bachelor’s degree in Biology, specialisation in virology.
Basic First Aid training course – 2018
Co-Leader & Medical Assistant
Miss Elizabeth MarlingBritishUniversity of Sussex. MSc Ecology, Biodiversity, and Conservation.Master’s degree in Ecology, Biodiversity, and Conservation, with a master’s thesis on bat ecology.
Bat Research Licence level 3 – 2019, experience with identifying, netting, and euthanising bats.
Research Assistant & Bat ID specialist
Mr Tobey ArthurBritishUniversity of Sussex. MSc Global Biodiversity Conservation.Master’s degree in Global Biodiversity Conservation, with a specialisation in rainforest habitats.Research Assistant
Dr Vincent MumfordBritishUniversity of Sussex. PhD Biology.PhD in Biology, with a specialisation in tropical diseases. Advanced First Aid training course – 2017Research Assistant & Chief Medical Officer
Dr Aaliyah CalathesBritishUniversity of Sussex. PhD Geography.PhD in Geography, with a specialisation in deforestation vs. degradation of rainforest habitats.Research Assistant
Professor Olayinka IsholaLiberianUniversity of Liberia. Head of Dept. Veterinary Public Health.Associate Professor, Tutor, and Lab Leader at the University of Liberia. Specialist in epidemiology, virology, and animal bacteriology.Team Leader of in-country members.

Professor Sampson CheaLiberianUniversity of Liberia. Tutor in Biochemistry.Assistant Professor, Tutor, and Lab Leader at the University of Liberia. Specialist in Biochemistry and Molecular Biology.Leader of Laboratory
Miss Sayamajunkon ForkayLiberianUniversity of Liberia. BSc BiologyBachelor’s degree in Biology. Published a significant paper on the Serological testing of Ebola. Student Leader in Professor Ishola’s Lab.Research Assistant
Miss Akeelah KamaraLiberianUniversity of Liberia. MSc Biosciences.Master’s degree in Biosciences, with a specialisation in medical science and virus testing.Research Assistant
Mr Mohammed KollieLiberianUniversity of Liberia. MSc Biology.Master’s degree in Biology, member of Professor Ishola’s Lab of Veterinary Public Health, specialisation in disease transmission & ecology.Research Assistant
Miss Islah FlomoLiberianUniversity of Liberia. BSc Zoology.Bachelor’s degree in Zoology, member of Professor Ishola’s Lab of Veterinary Public Health, Specialist knowledge of native bat species.Research Assistant

Budget

ExpendituresDescriptionPrice
Total
 
Pre-fieldwork / preparation

Transport to airport

 £120
Training
         
We have pre-trained bat, rainforest, and first aid specialists on the team. No training needed. £0
International travel

Flights are £420 each round trip x6

£2,520
Subsistence (accommodation and food)

 
First night at Johprin Motel in Harbel = £40
Bella Casa Hotel for our last 5 days, while working at the University Medical School = £920.
Food across the 22 days, for 6 people (based on estimates using Liberian Dollar) = £600
 £1,560
In-country travel.

4×4 van/coach, that fits 7 passengers from Jason Farmington Car Rental in Harbel is £130 per day
Petrol for 22 days: Fuel is £3 per gallon/ £0.60 per litre in Liberia.
A large van with OK fuel efficiency will use 10liters per 100km.
Our total travel is 209miles / 336km.
We can estimate a fuel cost of £50
 £2,910
 Local counterparts / guides
                        
N/A £0
Field equipmentWe will bring some light equipment from the UK, which we have permission to use from the University of Sussex: black-out bags, labelling for samples.
Other equipment will be lent to us by our associates at the University of Liberia: centrifuge for serology, liquid and dry nitrogen for sample preservation, test tubes and containers for samples. All analysis of samples will be carried out at the University’s facilities. We will need to purchase and bring new mist nets.
 £20
 InsuranceDue to the COVID-19 pandemic, all travel insurance companies, and comparison sites have suspended the services for international travel. We hope that this situation will have improved by November, when the study is due to begin, so an accurate insurance quote will be provided as soon as possible. £~

(estimate of £150)
 Medical / health & safety
         
Comprehensive first aid supplies £50
Post-fieldwork activitiesAnalysis of samples will be carried out in partnership with associates at University = £0
Drive hired van back to airport = £0 Journey Heathrow – Home = £120
£120
Preparation of project reportWriting report and leaflets for distribution= £0
Printing and Photocopying =£50
£50
Dissemination of findings          Review and publish report.£150
SUB-TOTAL £7,600
Contingency (usually 10% of sub-total) £706
TOTAL £8,306

Special Considerations

Environmental:
When mist netting for bats, we will have the nets up for as little time as possible, to avoid injuring or killing any other species; the nets will only be up during the night, bat’s active hours, and also the less conventionally active time for birds. We will not kill more bats than needed for effective analysis. All team members will be trained on environmentally friendly waste disposal.

Social & Cultural:
To avoid upset or alienation of local people, our presence on their land will be discussed prior to our arrival, and their voluntary involvement or assistance will be welcomed. All team members will be briefed on the local cultural norms before arrival. We will seek ethical and social approval for our methodologies before beginning, from our partners in Liberia.

Currently, as with many other countries around the world, Liberia has declared a state of emergency, due the COVID-19 pandemic. At present, estimates (using data from other countries that have moved past the worst infection/death rates) suggest that the lockdown measures in place will have lifted by mid-November, when the study is planned for. Others predict that a 2nd wave of the virus will develop in the winter, and these measures may have to be reinstated in certain countries. If this occurs, where travel is limited or the virus is extremely rife in Liberia, the study will be postponed.

However, if the COVID-19 crisis has passed, there are still advisories from the Government on travel to Liberia.
Travellers are advised to avoid large crowds such as demonstrations or places often visited by foreigners/tourists, to lessen the risk of terrorist attacks or crime such as robbery.
Many medical facilities expect to be paid up-front for treatment, so we must carry provisions for this, in case of emergencies. We must stay in contact with our host organisation about the support that they can provide to us while in the country.
We are told to avoid travelling at night outside Monrovia, except to or from Roberts International Airport. So, all of our driving will be done during daylight hours.

Risk Analysis & Crisis Management

If one of our team members was taken ill or injured whilst at either Base Camp (Lofa-Mano or Totota), they would be immediately assessed by our two medical officers (Dr Vincent Mumford and Miss Ruby Arthur), and an action plan would be made.
In the event of minor injuries or illness, such as sprains, cuts, bruises, dehydration, or heat exhaustion, the affected team member and medical officers will use the facilities onsite to treat the situation, and the patient will take the remainder of the day for rest, as a precaution, to avoid worsening the situation, or increasing probability of errors in the methodology.

In the event that a member of the team is seriously injured or becomes worryingly ill, the rest of the study will be called off, to ensure proper care of the individual.
If the patient is near our vehicle, we will pack up as much of the camp as possible.
If this occurs at Base Camp A, we drive to Liberia Government Hospital, which is the nearest health centre to Lofa-Mano (47 miles).
If this occurs at Base Camp B, we drive to the Phebe Hospital, which is the nearest health centre to Totota (31 miles).
If this occurs at our time working at the University, we will drive to the JFK Hospital in southern Monrovia (1.7 miles).
If a serious incident occurs and we are not nearby our vehicle or camp, the patient will be carried to the nearest access road, and an emergency vehicle will be contacted on our radio/ phone.
If a team member is taken to hospital with a serious injury, in-country members will contact relatives ASAP, and keep them updated on the treatment plan.

In the case of extreme circumstances, such as natural disasters or war, making it unsafe for the entire team to remain on location, we will follow the advice from the UK government in terms of an extraction plan. There is a British Embassy Centre in Monrovia, who will be made aware of our presence in the country, and we will have the direct contact details of a correspondent here. Local air evacuations operate out of Monrovia, and would be arranged if needed, through our long-distance radios, phones, or the emergency services.

Description & Maps of area

Environmental Conditions:
Liberia is on the southwest corner of the West Coast of Africa, bordering the Atlantic Ocean in the south, Sierra Leone in the west, Guinea in the north and Cote d’Ivoire in the east. Due to its equatorial position, a typical tropical weather system. The Liberian dry season lands typically December-February, meaning that our expedition will catch the end of the rainy season, so the plants and animals will be nourished and healthy, but we will avoid the threat of monsoons/flash floods.
The most remote area is the Lofa-Mano Base Camp, also the region posing the most risks to the study, facilities such as roads and electricity are less reliable. However, there are many small towns in the surrounding areas if we get lost or need to ask advice. The other areas of the expedition are well developed towns.

Cultural Conditions:
Our expedition involves travelling to various locations, we will come into contact with multiple ethnicities and groups, and the team will need in-depth training on conduct in these regions.
There are a number of cultural distinctions in Liberia, such as the high prevalence of ‘secret societies’, namely Poro, Sande, and Krahn. As well as the commonplace practices of polygamy, bushmeat hunting, and superstition: Liberian culture is characterised by a predisposition towards secrecy (encapsulated in the concept of ifa mo – “do not speak it”) and the involvement of mysterious forces in human lives.

References

1. Recent loss of closed forests is associated with Ebola virus disease outbreaks. Olivero, Jesus, Fa, John and Real, Raimundo. 14291, 2017, Scienific Reports , Vol. 7.

2. Lewnard, Joseph. Dynamics and control of Ebola virus transmission in Montserrado, Liberia: a mathematical modelling analysis. The Lancet: Infectious Diseases. 2014, Vol. 14, 12, pp. 1189-1195.

3. Fruit bats as reservoirs of Ebola virus. Leroy, Eric and al., et. s.l. : Nature, 2005, Nature , Vol. 438, pp. 575-576.

4. Weisse, Mikaela. When Tree Cover Loss is really Forest Loss: New Planation Maps Improve Forest Monitoring. Global Forest Watch. [Online] 26 January 2016. [Cited: 6 April 2020.] https://blog.globalforestwatch.org/data-and-research/when-tree-cover-loss-is-really-forest-loss-new-plantation-maps-improve-forest-monitoring.

5. Bowles, J. Ebola, jobs and economic activity in Liberia. Journal of Epidemiology & Community Health. 2016, Vol. 70, 1, pp. 271-277.

6. Overview, Control Strategies, and Lessons Learned in the CDC Response to the 2014–2016 Ebola Epidemic. Bell, B, Damon, I and Jernigan, D. 3, s.l. : Centres for Disease Control and Prevention, 2016, Vol. 65, pp. 4-11.

Published by amyandkatherine

We are two friends of 12 years, trying to start careers in journalism.

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