Running head: EBOLA VIRUS RESERVOIR 1

Race Against Viruses:

Chasing the Ebola Virus Reservoir in Democratic Republic of Congo

Annia I Niusulu

Hawaii Pacific University

EBOLA VIRUS RESERVOIR 2

SUMMARY

Throughout history, humanity has experienced a considerable number of lethal

epidemics. One of the latest global threat is known to be the Ebola virus, considered to be one of

the deadliest viruses encountered. The Ebola Virus Disease (EVD) is an extreme illness with a

high instance of casualty hazard (CFR). The Ebolavirus can be categorized into four species,

Ebola Sudan, E. Zaire, E. Ivory Coast and E. Reston. Ebola Sudan and E. Zaire are the most

widely recognized, on the account that they have shown the most outbreak in West Africa. After

review of past and current literature, it was revealed that the Ebolavirus is an animal-borne

(zoonotic) disease, transmitted from animals' hosts into humans. A great amount of studies has

been conducted to have a better understanding of the EVD epidemics. Many studies have been

conducted to test the hypothesis of African fruit bats as Ebola virus reservoir; thus, results remain

inconclusive due to the inability to isolate the viral strand from the bat species. However, there is

an alternate hypothesis that has yet to be explored, which prompted a rationale for investigating a

potential connection between Ebolavirus and insects that serve as food source for the bats.

Impeding primary contamination in humans alone would be the final victory in the battle against

EVD outbreaks. The focus of the study is to identify potential reservoirs of the Ebola virus; and

primary pathways in the spillover event between hosts, secondary vectors and humans of the

current population affected in the Kivu region; doing so that the government and other

international agencies can understand how to battle the threatening virus at its core.
EBOLA VIRUS RESERVOIR 3

INTRODUCTION

Emerging infections are an outgoing threat to species across the world. Terminal

pathogens such as a bacterium, fungi, and virus are considered to be the cause of many

pathogenic diseases. Humans and other beings are likely to become infected by pathogens that

enter the body through contact with other media. Once infected, organisms can continuously

spread the infection across communities, causing damaging outbreaks. The outcome of an

outbreak can vary based on the disease-causing pathogen, the size, and type of previous

outbreak, and the actual exposure to the disease (WHO, 2012). Deadly epidemics have been

circulating the globe for decades; for example, the Black Death, caused by the Bacteria Yersinia

pestis, that killed 75 million people in 1350 (CDC). Also, recent outbreaks such as the Avian

influenza A (H7N9) virus, Middle East respiratory syndrome coronavirus (MERS-CoV), the

Pandemic (H1N1) 2009, and the Ebolavirus, which is considered to be one of the deadliest

viruses encounter so far (WHO, 2012).

Although Ebola outbreaks are considered to be random, it is notable that the disease is

evolving exponentially, each time more potent. The first recorded Ebola virus disease (EVD)

outbreak, in a human population was around 1976-1979 in East Africa (Merens, Bigaillon,

Delaune, 2018). A more recent record of the deadliest reocurance happened between 2013-2016;

caused by drifting of the Ebola virus from Africa to places like Europe, and the United States.

Thus, producing a worldwide pandemic, unforeseen by public health organizations such as the

World Health Organization (WHO), and the Center for Disease Control and Prevention (CDC).

Numerous studies conducted have provided very few knowledges of the virus. Outcomes

point to several factors that impeded a rapid containment of the latest viral occurrence; aspects

such as shortage of health care, population mobility across borders, and the constant changing
EBOLA VIRUS RESERVOIR 4

and adapting of the virus genome (Stehling-Ariza, 2017). Subsequently, officials have

implemented plans worldwide for containment and battle against EVD outbreaks. Seemingly,

back in 2014, it was observed that the new outbreaks of the EVD were managed to be contained

in Lofa County, Liberia (Funk et al., 2017). The study concluded that having strong clinical and

community-based interventions in place can aid in successful containing the virus; however,

Ebolavirus reservoir still uncertain (Funk et al., 2017). Nonetheless, the problem remains, there

is no cure against the virus; EVD still stands as one of the deadliest infections.

While the Ebolavirus exists in other parts of the globe, it is in the African country where

is causing the most damage (Aftab, 2018). The latest outbreak begun on August of 2018, caused

by the E. Zaire species, considered the most fatal, with an overall case fatality ratio ranging from

69% to 88% (WHO, 2018). The CDC has reported cases in nine health zones in the Democratic

Republic of Congo. The Democratic Republic of Congo (DRC), is the southernmost country in

Central Africa, holding the fourth most populated area in Africa, and the sixteenth worldwide

(Maganga et al., 2014). The recent EVD outbreak in Congo has an eminent global danger not

only to humans but to flora and fauna as well (Barry et al., 2018). It is vital to assist DRC in

control and full elimination of. Though containment of EVD is the primary objective, the viral

reservoir has to be further investigated; this would aid in tracing the contamination pathway

between host-human. Impeding primary contamination in humans alone would be the final

victory in the battle against EVD outbreaks.

BACKGROUND

Ebola virus is a member of the Filoviridae family, which includes three genera:

Cuevavirus, Marburgvirus, and Ebolavirus. Within genus Ebolavirus, there are four major
EBOLA VIRUS RESERVOIR 5

groups known to cause disease in humans. The four species are Ebola Sudan, E. Zaire, E. Ivory

Coast and E. Reston (Merens, Bigaillon, Delaune, 2018). The RNA of the Ebolavirus is an

enveloped, non-segmented and negative-stranded; viral particles have shown presence of viral

nucleocapsid in the form of a helical single-stranded RNA genome wrapped around viral proteins

NP, VP35, VP30, and L (Malashkevich et al., 1999). Ebolavirus is an animal-borne (zoonotic)

disease, transmitted from animals' hosts into humans. Hence, Ebolavirus is believed to spread

from its host reservoir into humans by consumption or handling infected animals (Rewar &

Mirdha, 2015). Further, EVD spreads via human-to-human interactions, which is considered to

be the primary source of EVD outbreaks in populations (Merens, Bigaillon, Delaune, 2018). In

addition, Figure 1 shows EVD contamination cascade and effects in human organs as illustrated

by previous studies conducted in 2015 by Gebretadik, Seifu, & Gelaw, B.

Still, there is much to be explored; the ecology of the EVD outbreaks, and more

importantly, the specific reservoir of the Ebolavirus is still undetermined. The support for fruit

bats as main reservoirs has been explored in various studies (Arinjay et al., 2018). For example,

in 1996, a study was conducted by Swanepoel et al. with a diverse species of plants and animals,

to test for possible reservoirs of Ebolavirus. During the experiment, the strand, E. Zaire, was

introduced into plants and animal species. The results showed that the virus could be isolated

from bat species Tadarida condylura, T. pumila and Epomophorus wahlbergi, following virus

replication and seroconversion (Swanepoel et al., 1996). Additionally, the results suggested that

the bat species infected with the virus showed no clinical illness post infection; supporting the

idea that the Ebolavirus does not harm bats, making them potential hosts (Swanepoel et al.,

1996). Conversely, results were unable to provide strong evidence supporting the further

transmission of the Ebola virus from bats to another organism (Swanepoel et al., 1996).
EBOLA VIRUS RESERVOIR 6

In another study conducted in 2005, scientist Eric M. Leroy presented molecular evidence

that characterized African fruit bats as the primary reservoir hosts for the Ebolavirus (Alexandre

et al., 2016). In the analysis, numerous samples of birds, bats, and small vertebrates, taken from

infected sites, were tested for the presence of Filoviruses. The results showed the presence of

Marburgvirus and Ebolavirus. From genus Ebolavirus, species E. Sudan, and E. Zaire were

identified by the existence of specific molecule for the Ebolavirus. Immunoglobulin G was found

in 4 out of 17 Hypsignathus monstrous, 8 of 117 Epomops franqueti and 4 out of 58 Myonycteris

torquata bat species (Leroy et al., 2005). Also, the relationship was established between the three

species and their habitat around Ebolavirus hotspots. However, the study was inconsistent; PCR

analysis showed small presence of the Ebolavirus RNA, revealing the possibility that bats were

infected from another host (Leroy et al., 2005).

Critically, during this review it has been observed that many studies have shown

favorable results in the correlation between African fruit bats and the Marburgvirus genus

(Amman et al., 2012; Paweska et al., 2012); however, there are many irregularities observed in

the studies of fruit bats. There is a need to search deeper into the possibility that fruit bats can

become contaminated from other sources. Due to the lack of reliability, a review of current

literature will be conducted to gain a better understanding about the reservoirs of the Ebolavirus

in the Democratic Republic of Congo.

LITERATURE REVIEW

Since its emergence in the African continent in 1976, Ebolavirus has been the center of

many questions. The hypothesis surrounding fruit bats as a potential viral reservoir has been the

main focus of many studies but remains inconclusive. Also, spillover events from EVD
EBOLA VIRUS RESERVOIR 7

reservoirs to human vectors is still unclear. Following is an analysis of current literature over the

natural and epidemiological pathways of the EVD reoccurrence.

Bats Species as Reservoir of the Ebola Virus

Bats have been known to be the cause of several human diseases such as Rabies virus,

Severe Acute Respiratory Syndrome-related Coronavirus (SARS-CoV) and others (Smith and

Wang, 2013). Bats belong to the order Chiroptera, and have been the center of studies for

decades, in order to identify Filoviruses reservoirs in Africa. The hypothesis behind bats as the

main reservoir for filoviruses lays in that, when infected, bats do not show symptoms of illness.

During the review, it was noted that most studies use similar methods for research; such as

setting bat traps at the viral hotspots, for a determined amount of time (Pourrut et al.; Paweska et

al., 2012). Consequently, the majority of the focused on analysis of the collected blood

specimens from the bat species; also, the molecular amplification of extracted DNA using

polymerase chain reaction (PCR) examination.

The first viral sequence isolated from bats was during the EVD outbreak in Gabon in

2003 (Pourrut et al., 2007). The results revealed possible E. Zaire reservoirs in about 67% of the

2070 bats capture (Pourrut et al., 2007). The viral RNA 13 appeared in specimens of Epomops

franqueti, Hypsignathus monstrous, and Myonycteris torquata. Additionally, 16 samples

contained EBOV- specific antibodies, identified through blood samples (Pourrut et al., 2007).

Furthermore, Table 1 and Table 2 shows results from the study conducted by Pourrut et al. on

the presence of Ebola antibodies and its prevalence in bat species, before and after Ebola

outbreaks in Gabon and DRC. The results exposed a significant variance in IgG antibody

prevalence of the E. Zaire strand between species of bats collected during the outbreak period

and those collected after the outbreak period.

EBOLA VIRUS RESERVOIR 8

Moreover, in a study conducted in 2012, Marburgvirus viral strand showed in multiple

tissues 2-9 days post-infection, in the Egyptian fruit bat species R. aegyptiacus (Paweska et al.,

2012). Also, 9 to 21 days post infection; the bat species showed IgG antibodies present. Results

also revealed that neither of the bat’s species tested in the study by Paweska et al. showed

clinical symptoms post infection; supporting the idea of bats as Ebolavirus hosts. However, both

studies were flawed. For example, Paweska et al. were unable to isolate the virus from bats

feces. The data was unsupported of the idea that viral transmission can be originated from the

secretion of infected bats (Paweska et al., 2012). Secondly, in the study conducted by Pourrut et

al., it was concluded that the sizes of the samples were too little too drawn out meaningful

conclusions.

In a more recent study, in 2018, Goldstein et al. discovered a new strand of Ebolavirus in

Sierra Leone. Given the name, the Bombali virus, it was found in bat species Chaerephon

pumilus and Mops condylurus. According to the scientists, the bat species were found carrying

viral molecules that can invade human cells. Besides, the report explains how the bats were

found roosting inside homes within the local community; supporting the idea of bats as Ebola

virus’s reservoir. Also, the results supported the pathway of EVD transmission from bats to

humans. However, Goldstein and associates also indicated that further studies about the virus

specifics are necessary; primarily around the area the virus was found.

Conclusively, during the review of several works of literature, it was noted potential

connections between gender, males to female ratio, offspring survival, migration of bats, and

deforestation, as factors in need of further research and clarification. Besides, the literature

reviewed did not provide any information towards studies focusing on insects that are part of

bat's diets as a possible reservoir for the virus. Filovirus ecology, in specially, the specific
EBOLA VIRUS RESERVOIR 9

relations between bats and the Ebolavirus are still unclear. Further investigation is still needed in

order to determine the role of bats in the Ebola virus contamination pathway.

Non-Human Vectors of Ebola virus

While the idea still surrounds bats as primary sources of Ebolavirus, other animals can be

vectors of the deadly disease. Primates, for example, were believed to be the cause of infections;

the idea of primates as reservoirs was unsupported due to the high rate of mortality following

infection. However, it has been supported that primates do participate in the spillover event into

human populations (Harish et al., 2015) Besides, several studies have also suggested that

surveillance of potential Ebola outbreaks within the local population of primates can serve as a

method of preventing human contamination.

Furthermore in 2005, Rouquet et al. published a study in which the results point to a

correlation between the EVD outbreaks in animals and humans. During the study, the scientists

identified species of animal carcasses to include, Gorillas, and Chimpanzees as victims of the

virus before the outbreak spilled into the human population. In addition, results from the study as

seen in Figure 2, showed the distribution of carcasses found within the DRC and Gabon borders

surrounding the EVD outbreak in human population between 2001-2003 (Rouquet et al., 2015).

Seemingly, in a separate study conducted by scientists Pigott et al. the results also suggested a

correlation between animals infected with the Ebolavirus and outbreak in human population.

Also, the scientists reported that a significant infection in primates was found to be around the

DRC, which can be observed in Table 3, and Table 4; the information was taken from the study

and provides data from between 1994-2008 on the location and types of animals found affected

by the Ebola disease, using reported data of Ebola virus in animals as of 2014.
EBOLA VIRUS RESERVOIR 10

CONCLUSION

The Democratic Republic of Congo has been the site of numerous studies related to the

Ebolavirus disease since 1976. Furthermore, with the most recent EVD outbreak, in 2018, the

Democratic Republic of Congo stands as an eminent global danger not only to humans but to

flora and fauna as well. The Democratic Republic of Congo is home to many rare and

endangered species such as the Bonobo and the Okapi; it is of high urgency to stop these species

from further contamination and to do so, it is vital to investigate Ebola virus reservoirs.

The conclusion of all of the studies reviewed indicated that, although, there has been

extensive research surrounding Ebolavirus and its spread in the African continent, preventing

primary infections between host and humans is still unresolved. Furthermore, the ecology and

main reservoir of the Ebolavirus remains undetermined. It is vital to assist DRC in containment

and full elimination of the Ebolavirus. In order to do so, further investigation on how the virus

persists and reemerges in the DRC is fundamental. This document proposes to investigate one of

the nine health zones in DRC with the most current EVD outbreak. Furthermore, the study will

focus on identifying the factors involved in the spillover event, in order to locate the Ebolavirus

reservoir and secondary vectors.

RATIONALE

After reviewing much of the past and current literature, it is noted that many of the

studies were inconclusive and presented many flaws. Consequently, it is clear that there are little

to no conclusive answers and sound evidence pointing to the reservoir for the Ebolavirus in the
EBOLA VIRUS RESERVOIR 11

DRC. Also, it is apparent that there are many unanswered questions as to why the Ebolavirus

keeps resurfacing in the DRC.

The spread of Ebolavirus disease, in 2013-2016 to other parts of the globe, placed the

planet in a state of emergency unforeseen by many officials, and the general population. The

fashion in which Ebolavirus keeps emerging in DRC is a gamble that humanity cannot afford to

take. The country’s proximity to regional borders constitutes a risk for further contamination.

Also, the high migration status, from mine workers, and asylum seekers, to the many natives

that transit to South Africa and Europe, deduces a substantially high risk of another global

spread. The Democratic Republic of Congo stands as the home to the second largest rainforest in

the world, making it the most biodiverse African country. Thus, the country’s possession of rare

and endangered species such as the Bonobo, and the Okapi; raises preservation worries.

The main reason for investigating the current EVD is because the Democratic Republic

of Congo stands as an eminent, large-scale threat not only to humans but to flora and fauna as

well. Similarly, if left unchecked the virus can quickly be spread to the other parts of the

continent and finally to the whole world. In other words, it is easy for Ebola to become an

international epidemic if left unsolved. Many studies have been conducted to test the hypothesis

of African fruit bats as Ebolavirus reservoir; thus, results remain inconclusive due to the inability

to isolate the viral strand. However, there is an alternate hypothesis that has yet to be explored

and prompted the rationale for investigating a potential connection between Ebolavirus and

insects that serve as food source for the bats. The focus of the study is to identify potential

reservoirs of the Ebola virus; and primary pathways in the spillover event between hosts,

secondary vectors and human population affected in the Kivu region; doing so that the
EBOLA VIRUS RESERVOIR 12

government and other international agencies can understand how to combat the dangerous virus

at its core.

METHODS

A team of 15 scientists will deploy to the Democratic Republic of Congo. Exclusively, to

the Beni providence located in North Kivu, where CDC reports the latest Ebola virus outbreak

started in October of 2018. The methods employed by the team will be as followed:

• Identify 10 collection sites around the Beni Providence determined by Ebola outbreak in

humans.

• Set up bat traps in each site and collect samples for 20 days at each location.

• Following capture, bats will be sorted into different cages by species.

• Collect blood, urine and/or feces samples from all specimens

• Isolated samples by

- Bat Species

- Sex (M/F)

- Ages (0-5, 6-11, 12-17, 18-20 years).

• After samples are taken, bats are to be released into their habitat.

Consequently, local insects that are known as the food source for the bat species collected will

also be gathered and samples of blood will be collected and sorted out by species and collection

site. The scientific team will be divided into teams:

• Team 1: Scientist 1-5 will be in charge of setting up and monitoring collection sites for

the bat species. Will also be sorting the bat species according to the sorting methods
EBOLA VIRUS RESERVOIR 13

explained above. Team 1 will also be responsible for releasing bats after use and for

retracting the traps.

• Team 2: Scientist 6-10 will be responsible for collecting local insect species known as the

food source for the local bat species. Will also sort insects according to species.

• Team 3: Scientist 11-15 will be responsible for collecting blood, urine and /or feces

samples from bats species in the appropriate vial and preparing them for transport to a

medical laboratory in the United States.

Samples from both, bats species and insect species, will be brought back to the laboratory

in the United States. The tests will be conducted using Enzyme-linked immunosorbent assay

(ELISA), and Reverse transcriptase polymerase chain reaction (PCR). The main focus during

testing will be to isolate the Ebola virus RNA from the species collected. The results from this

study will be compared to data from existing human and nonhuman victims of EVD, in the North

Kivu region. The goal is to stablish a contamination pathway from insects to bats, to human and

nonhuman vectors. According to the results, models such as tables and figures will constructed.

TIME TABLE

Below is the log format to be utilize in this study. A new time table will be used for each

of the 10 viral hotspots that will be identify once in country. The dates will be adjusted

accordingly. The length of the study is expected to be 320 days.

Date Day Activities

2/11/19 1st Arriving in Kivu province.
EBOLA VIRUS RESERVOIR 14

Setting up base camp and locating potential collection sites where

Team 1 and 2 will be setting up their camp.

3/11/19 2nd Team 1 and 2 will deploy to collection sites and set up traps.
Team 3 will remain at main camp and set up for specimen collection.

4/11/2019- 20 days Collection day begins.

24/11/2019
Team 1 will monitor, collect and sort bat species
Team 2 will also begin to monitor, collect and sort insect species
Team 3 will support 1 and 2 by providing meals and beverages and
any other needs in the daily basics.

24/11/2019- 10 days All species will be brought back to main camp.

3/12/2019
Team 3 will then collect urine, blood and feces samples from all bat
and insect species. Team 3 will also prepare vials for transport to the
laboratory.
Team 1 and 2 will commence releasing bats into their habitats as Team
1 is done conducting their part.
Clean up and packing will be conducted by all teams.
All teams will move to next hotspot

EXPECTED OUTCOME

Primary, tests result from the bat samples collected is expected to provide information on the

type of Ebola viral species, the viral strength differences between females and males, and viral

strand differences between bats in the ages of 0-5, 6-11, 12-17, 18-20 years. Additionally, test

results from samples collected from the insects are expected to show whether the presence of
EBOLA VIRUS RESERVOIR 15

Ebola is observed or not. Precisely, Ebola virus is highly expected to be present in the blood of

some species of the insects collected.

Following, results from PCR and Immunoglobin tests form the samples obtain from the

bat's species will be compared to the database containing the results from the database of blood

samples obtained from the human and nonhuman populations, currently infected with the Ebola

virus in the Beni providence. It is expected to create a relationship between the viral RNA

strands found in the different bat species to those of the human and nonhuman populations.

Hence this relationship will be critical to trace spillover events from bats to nonhuman

populations to human populations.

SIGNIFICANCE

If accepted, the hypothesis that Ebola virus reservoir can be found in insects that are used

for consumption, can be a huge breakthrough in the fight against this deadly disease. Data from

this study can excite researches to take an ecological view and critically reconsider and review if

there are other factors that influence filovirus disease outbreak patterns. Determining the virus

reservoir will not only help answer questions about how the virus emerges, but also help local

and international public health workers to be more prepared and aware of future epidemic risks.

TABLES AND FIGURES

EBOLA VIRUS RESERVOIR 16

Figure 1- Taxonomy of filovirus as illustrated previous studies conducted in 2015 by

Gebretadik, Seifu, & Gelaw, B.
EBOLA VIRUS RESERVOIR 17

Figure 2- Geographical representation of potential bat reservoirs for the Ebola virus.
Taken from Olivar and Hayman, 2014.

Table 1. Table taken from studies conducted by Pourrut et al., 2007.

EBOLA VIRUS RESERVOIR 18

Table 2. Table taken from studies conducted by Pourrut et al., 2007.

EBOLA VIRUS RESERVOIR 19

Table 3- Displays information from 1994-2008 on location and types of animals found
affected by the Ebola disease. The table was taken from another study and referenced
(Pigott et al., 2014)
EBOLA VIRUS RESERVOIR 20

Table 4- Displays information from 1994-2008 on location and types of animals found
affected by the Ebola disease. The table was taken from another study and referenced
(Pigott et al., 2014)
EBOLA VIRUS RESERVOIR 21

References

Amman, B. R., Carroll, S. A., Reed, Z. D., Sealy, T. K., Balinandi, S., Swanepoel, R., ... &

Cannon, D. L. (2012). Seasonal pulses of Marburg virus circulation in juvenile Rousettus

aegyptiacus bats coincide with periods of increased risk of human infection. PLoS

Pathogens, 8(10), e1002877.

Aftab A. Ansari. (2018). Clinical features and pathobiology of Ebolavirus infection, Journal of

Autoimmunity. Volume 55, ISSN 0896-8411, https://doi.org/10.1016/j.jaut.2014.09.001.

Alexandre Hassanin, Nicolas Nesi, Julie Marin, Blaise Kadjo, Xavier Pourrut, Éric Leroy, Guy-

Crispin Gembu, Prescott Musaba Aikawa, Carine Ngoagouni, Emmanuel Nakouné,

Manuel Ruedi, Didier Tshikung, Célestin Pongombo Shongo, Céline Bonillo. (2016).

Comparative phylogeography of African fruit bats (Chiroptera, Pteropodidae) provide

new insights into the outbreak of Ebola virus disease in West Africa, 2014–2016.

Comptes Rendus Biologies, ISSN 1631-0691, https://doi.org/10.1016/j.crvi.2016.09.005.

Arinjay Banerjee, Vikram Misra, Tony Schountz, Michelle L. Baker. (2018). Tools to study

pathogen-host interactions in bats. Virus Research. Volume 248, ISSN 0168-1702,

https://doi.org/10.1016/j.virusres.2018.02.013.
EBOLA VIRUS RESERVOIR 22

Barry, A., Ahuka-Mundeke, S., Ahmed, Y. A., Allarangar, Y., Anoko, J., Archer, B. N., ... &

Bokenge, T. (2018). Outbreak of Ebola virus disease in the Democratic Republic of the

Congo, April–May 2018: an epidemiological study. The Lancet, 392(10143), 213-221.

Disease outbreaks. (2012, August 24). Retrieved from

http://www.who.int/environmental_health_emergencies/disease_outbreaks/en/

Funk, S., Ciglenecki, I., Tiffany, A., Gignoux, E., Camacho, A., Eggo, R. M., Kucharski, A. J.,

Edmunds, W. J., Bolongei, J., Azuma, P., Clement, P., Alpha, T. S., Sterk, E., Telfer, B.,

Engel, G., Parker, L. A., Suzuki, M., Heijenberg, N., … Reeder, B. (2017). The impact of

control strategies and behavioural changes on the elimination of Ebola from Lofa County,

Liberia. Philosophical transactions of the Royal Society of London. Series B, Biological

sciences, 372(1721), 20160302.

Gebretadik, F. A., Seifu, M. F., & Gelaw, B. K. (2015). Review on Ebola virus disease: its

outbreak and current status. Epidemiology (sunnyvale), 5(204), 2161-1165.

Goldstein, T., Anthony, S. J., Gbakima, A., Bird, B. H., Bangura, J., Tremeau-Bravard, A., ... &

Grodus, M. (2018). The discovery of Bombali virus adds further support for bats as hosts

of ebolaviruses. Nature microbiology, 3(10), 1084.

EBOLA VIRUS RESERVOIR 23

Harish Rajak, Deepak Kumar Jain, Avineesh Singh, Ajay Kumar Sharma, Anshuman Dixit.

(2015). Ebola virus disease: past, present, and future. Asian Pacific Journal of Tropical

Biomedicine. Volume 5, Issue 5. Pages 337-343. ISSN 2221-1691.

https://doi.org/10.1016/S2221-1691(15)30365-8.

Leroy, E. M., Epelboin, A., Mondonge, V., Pourrut, X., Gonzalez, J. P., Muyembe-Tamfum, J. J.,

& Formenty, P. (2009). Human Ebola outbreak resulting from direct exposure to fruit bats

in Luebo, Democratic Republic of Congo, 2007. Vector-borne and zoonotic

diseases, 9(6), 723-728.

Leroy, E. M., Kumulungui, B., Pourrut, X., Rouquet, P., Hassanin, A., Yaba, P., ... & Swanepoel,

R. (2005). Fruit bats as reservoirs of Ebola virus. Nature, 438(7068), 575.

Maganga, G. D., Kapetshi, J., Berthet, N., Kebela Ilunga, B., Kabange, F., Mbala Kingebeni,

P., ... & Cabore, J. (2014). Ebola virus disease in the Democratic Republic of Congo.

New England Journal of Medicine, 371(22), 2083-2091.

Malashkevich, V. N., Schneider, B. J., McNally, M. L., Milhollen, M. A., Pang, J. X., & Kim, P.

S. (1999). Core structure of the envelope glycoprotein GP2 from Ebola virus at 1.9-Å

resolution. Proceedings of the National Academy of Sciences, 96(6), 2662-2667.

EBOLA VIRUS RESERVOIR 24

Mérens, A., Bigaillon, C., & Delaune, D. (2018). Ebola virus disease: Biological and diagnostic

evolution from 2014 to 2017. Médecine Et Maladies Infectieuses, 48(2), 83-94.

doi:10.1016/j.medmal.2017.11.002

Narra, R., Sobel, J., Piper, C., Gould, D., Bhadelia, N., Dott, M., … Jhung, M. (2017). CDC

Safety Training Course for Ebola Virus Disease Healthcare Workers. Emerging Infectious

Diseases, 23, S217–S224. https://doi-org.hpu.idm.oclc.org/10.3201/eid2313.170549

Paweska, J. T., Van Vuren, P. J., Masumu, J., Leman, P. A., Grobbelaar, A. A., Birkhead, M., ... &

Kemp, A. (2012). Virological and serological findings in Rousettus aegyptiacus

experimentally inoculated with vero cells-adapted hogan strain of Marburg virus. PloS

one, 7(9), e45479.

Pigott, D. M., Golding, N., Mylne, A., Huang, Z., Henry, A. J., Weiss, D. J., Brady, O. J.,

Kraemer, M. U., Smith, D. L., Moyes, C. L., Bhatt, S., Gething, P. W., Horby, P. W.,

Bogoch, I. I., Brownstein, J. S., Mekaru, S. R., Tatem, A. J., Khan, K., … Hay, S. I.

(2014). Mapping the zoonotic niche of Ebola virus disease in Africa. eLife, 3, e04395.

doi:10.7554/eLife.04395

Pigott, D. M., Millear, A. I., Earl, L., Morozoff, C., Han, B. A., Shearer, F. M., ... & Bhatt, S.

(2016). Updates to the zoonotic niche map of Ebola virus disease in Africa. Elife, 5,

e16412.
EBOLA VIRUS RESERVOIR 25

Pourrut, X., Delicat, A., Rollin, P. E., Ksiazek, T. G., Gonzalez, J. P., & Leroy, E. M. (2007).

Spatial and temporal patterns of Zaire ebolavirus antibody prevalence in the possible

reservoir bat species. The Journal of infectious diseases, 196(Supplement_2), S176-S183

Pourrut, X., Kumulungui, B., Wittmann, T., Moussavou, G., Délicat, A., Yaba, P., ... & Leroy, E.

M. (2005). The natural history of Ebola virus in Africa. Microbes and infection, 7(7-8),

1005-1014.

Rewar, S., & Mirdha, D. (2015). Transmission of Ebola Virus Disease: An Overview. Annals of

Global Health, 80(6), 444. doi:10.1016/j. aogh.2015.02.005

Rouquet, P., Froment, J. M., Bermejo, M., Kilbourn, A., Karesh, W., Reed, P., Kumulungui, B.,

Yaba, P., Délicat, A., Rollin, P. E., … Leroy, E. M. (2005). Wild animal mortality

monitoring and human Ebola outbreaks, Gabon and Republic of Congo, 2001-2003.

Emerging infectious diseases, 11(2), 283-90.

Smith, I., & Wang, L. F. (2013). Bats and their virome: an important source of emerging viruses

capable of infecting humans. Current opinion in virology, 3(1), 84-91.

EBOLA VIRUS RESERVOIR 26

Stehling-Ariza, T., Lefevre, A., Calles, D., Djawe, K., Garfield, R., Gerber, M., … Shahpar, C.

(2017). Establishment of CDC Global Rapid Response Team to Ensure Global Health

Security. Emerging Infectious Diseases, 23, S203–S209. https://doi-

org.hpu.idm.oclc.org/10.3201/eid2313.170711

Smith, D. W., Rawlinson, W. D., Kok, J., Dwyer, D. E., & Catton, M. (2015). Virological

diagnosis of Ebolavirus infection. Pathology, 47(5), 410-3.

Swanepoel, R., Leman, P. A., Burt, F. J., Zachariades, N. A., Braack, L. E., Ksiazek, T. G.,

Rollin, P. E., Zaki, S. R., … Peters, C. J. (1996). Experimental inoculation of plants and

animals with Ebola virus. Emerging infectious diseases, 2(4), 321-5.

WHO. (2018). Ebola Virus Disease Factsheet.

www.who.int/news-room/fact-sheets/detail/ebola-virus-disease