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Leakey Foundation Final Report Survey of bonobos (Pan paniscus) in Tshuapa-Lomami-Lualaba area, Democratic Republic of Congo Paco Bertolani Department of Archaeology and Anthropology University of Cambridge Brief Summary The main objective of this pilot study was to assess the feasibility of establishing a long-term site for bonobo behavioural research in TL2 region, Maniema province, Democratic Republic of the Congo. During a 3-month field trip, I was able to spend two full months in the forest looking for bonobos and collecting faecal samples for DNA analysis. The other month was mostly used for travelling to and from the site. As expected, bonobos were difficult to find, and not being habituated, whenever they were found, they ran away giving alarm calls. We tried to track them, but with little success. Nevertheless, we were able to see them 7 times on different days. From October to December 2012, we collected 87 fresh faecal samples, and an additional 69 samples were collected by field assistants between January and March 2013, thanks to funding provided by Dr. Beatrice Hahn, University of Pennsylvania. From these 156 faecal samples, we extracted bonobo DNA from 128. A preliminary analysis of mitochondrial DNA and 8 microsatellite nuclear loci revealed a minimum number of 63 individual bonobos. Poachers regularly visit the area and hunt bonobos and other wildlife. Such illegal hunting is not much by the local communities bordering the park because the TL2 project seemed to be successful in persuading the local people. Despite the remoteness of the site, and the presence of hunters, I believe that establishing a long-term research site in TL2 is feasible and recommended for the potential interesting results and the positive effect on bonobo conservation in the long-term. Brief Summary of Publications The 128 bonobo faecal samples collected during this study contributed to a large scale project on the origin of the Plasmodium vivax parasite. This study was recently published in a collaborative paper: Liu et al. 2014. African origin of the malaria parasite Plasmodium vivax. Nature Communications 5:3346. DOI: 10.1038/ncomms4346. As expected, all bonobo samples were negative from the parasite, but the samples collected from other apes across their African distribution, and the sequence of the Plasmodium vivax have been interpreted in light of an

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African origin of P. vivax parasite, against the previous assumption that P. vivax was a Southeast Asian species. For the genetic characterisation of the individuals sampled, additional analyses are needed in order to know how many samples are duplicat4es (or not). I am currently collaborating with geneticists in the University of Cambridge and the Sanger Institute to further analyse the DNA samples collected in the field. Such a work will potentially lead to additional publications. Detailed description of results Study site The site is very remote, accessible from the town of Kindu (capital of Maniema province) by a two-day trip, first day on motorbike to the village of Chombe Kilima, and second day on foot for about 45 km along a path crossing the proposed TL2 national park. This trail leads to Katopa, a village just outside the protected area on the western bank of Lomami River (Figure 1). Katopa is one of the main permanent camps of TL2 Project.

Figure 1: TL2 proposed National Park in the Democratic Republic of Congo.

Map of the study site In order to navigate through the site and place bonobo observations in a geographic context, I did a GIS map of the study area using remote sensing datasets and data collected on the ground with GPS. A few versions of this map are illustrated in Figures 1, 2, and 5.

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Figure 2: Study area in TL2.

Bonobo sightings and traces With a field team of four persons, based in two temporary camps, we were able to survey an area covering about 75 km2. We encountered traces of bonobos (old nests and feeding remains) almost everywhere we went (Figure 2). Over two months, we were able to see bonobos seven times. Their reaction upon seeing humans was the typical reaction of non-habituated apes: alarm calls and fleeing, then becoming silent.

Figure 3: Bonobos found at their nest site early in the morning. Right: Adolescent male; left: adult female with infant.

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Genetics We collected a total of 156 fresh faecal samples from which we could extract bonobo DNA in 128 of them. 124 samples were from the area surveyed during this study, other 32 were collected opportunistically outside the surveyed area, and some of these (N=15) were from the other side of the Lomami river (Figure 5). The 128 samples with enough bonobo DNA had 16 different mitochondrial haplotypes (D-loop). The first preliminary analysis of 8 microsatellite loci in different chromosomes revealed the presence of at least 63 individuals. The real number of individuals is likely to be higher because of the presence of null alleles, which can represent either homozygous at that locus, or heterozygous in which PCR (Polymerase Chain Reaction) failed to amplify one allele. The presence of more individuals is confirmed by the fact that the same genetic profile was found in places too far away to be visited by the same individual, and because samples from different individuals (collected at the same site, but from different nests) had the same genetic profile. It is also possible that two individuals share the same allele at each of the 8 loci analysed. In addition, bonobos are not strictly territorial like their sister species, the chimpanzee. Although each community has its own territory, it has been observed that individuals of two different communities can spend some time together (up to a few days). Thus, samples collected at one place not necessarily belong to individuals from the same community. For these reasons, the first analysis of the genetic data collected cannot tell us how many communities are in the areas surveyed, and how big are these communities. Figure 4: Collection of faecal samples. Right: Jean Serge Ikembe (field assistant); left: myself.

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Figure 5: Sites where fresh faecal samples were found.

Figure 6: Faecal samples collected from two different nesting sites on a single day (N=20).

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Illegal Hunting Poaching is a serious issue in TL2 proposed National Park. Although the efforts of the TL2 conservation project were successful in decreasing the hunting pressure by people living in the surrounding areas, some hunting still occurs by “professional” hunters from more distant areas. Traces of hunting activity, such as old camps or cartridges are abundant. In the past, hunters have burnt TL2 camps and left messages written on the trees (Figure 7). The project also burns poachers’ camps (and meat if found) whenever they are encountered. If shotguns are heard by the project team and collaborators, an expedition is organised in order to catch the poachers, together with authorities of ICCN (Institut Congolais pour la Conservation et la Nature). During my stay in TL2, two captures occurred. The first time a poacher was caught, and was kept for 3 days in the village (Katopa), “judged” by the chef du village and other people, his rifle and hunting dogs confiscated, and he was subsequently released. The second episode was more serious and led to severe injuries by a member of the TL2 project, as well as the poacher. This time the poacher was taken to court in Kindu. A report of this episode is described in the following website: http://www.bonoboincongo.com/2012/10/22/ambush-by-armed-poachers-in-congo/

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A C Figure 7: A) Message left by poachers at Camp bonobo, after having burnt it. From Kiswahili (badly written): “TL2, there is no park here. If you dare to come here, you will die. Fuck off”. B) Smoked bonobo hands found by TL2 project staff at a poachers’ camp. C) A temporary poachers’ camp.

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Conclusions Poaching in TL2 is a serious problem, but bonobos and other wildlife are still abundant. Establishing a long-term research site is feasible, although risky. Over the long term it might have a protective effect on wildlife, discouraging poachers from visiting the nearby areas. This is more or less what happened in Taï National Park, Ivory Coast, where the permanent research station has discouraged poachers from hunting in the areas near the research station, with a visible effect of wildlife densities, which are higher in the vicinity of the research station than anywhere else in the National Park. Acknowledgements I thank the Leakey Foundation and Beatrice Hahn for funding to conduct this pilot study. ICCN (Institut Congolais pour la Conservation et la Nature) for granting permits to work in DRC. I am grateful to Terese and John Hart, and all the staff of TL2 project for logistic collaboration in DRC. Special thanks to our field team: Jean Serge, Jamais Tombé, and Brazos.