Using biodiversity surveys, website and film to promote the value of the Kabobo massif for conservation support. Final Report

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1 Using biodiversity surveys, website and film to promote the value of the Kabobo massif for conservation support Final Report

2 Contents Project outputs/results... 3 Biodiversity survey reports... 3 Kabobo Massif Profile... 3 Kabobo Massif Biodiversity... 4 Current conservation challenges... 5 On site DNA sequencing... 6 Technology... 6 Preliminary tests... 6 Kabobo Massif test... 6 Conclusion... 7 Mammals... 8 Large mammals checklist Small mammals (non bats) checklist Amphibians and Reptiles Amphibians checklist Reptiles checklist References CEPF SMALL GRANT FINAL PROJECT COMPLETION REPORT Short-term impacts (project objectives) Long-term impacts (project objectives) Lessons Learned Additional Funding Safeguard Policy Assessment Additional Comments/Recommendations... 22

3 Project outputs/results Michele Menegon, Ana Rodriguez Prieto, Marina Moreno, Deo Kujirakwinja Biodiversity survey reports The project is part of the on-going collaboration between the Science Museum of Trento and Wildlife Conservation Society (WCS) DR Congo Program. All the activities in the field and the subsequent work of reporting and film production have been carried out in close collaboration. The main aim of the project was to collect scientific and visual information on Kabobo massif in order to highlight its biological value and to stress the need of a comprehensive, effective conservation strategy for the whole area. To target areas geographically distant from those previously surveyed and cover places never before sampled, in particular to investigate reptiles and amphibians and collect the genetic material needed to assess the actual herpetological diversity. During our expedition, we also carried out on site DNA sequencing to species belonging to different taxonomic groups, through the use of a mobile lab that can perform DNA extraction, amplification and sequencing outside a laboratory (see The obtained sequences where then sent to GenBank via satellite connection, to verify the genetic identity of the sample. The expedition and main features and Kabobo landscape and beauty have been the subject of a documentary and a dedicated real-time blog and webpage. Note that due to custom issues, only recently solved, the samples collected during the 2017 expedition have yet to be exported to Europe for more detailed analysis. Kabobo Massif Profile The Kabobo massif is the largest, isolated area of montane forest in Eastern DRC, it stretches for about 100 km along the western shore of the Lake Tanganyika and is naturally separated from the Itombwe massif to the north, by a 50 km wide savannah through which crosses the Kilombwe River (Kerbis et al., 2013), making it an area of potential high species endemism, as the savannah could constitutes a barrier to dispersal both for forest associated bird, mammal, reptile and amphibian species. Given its size, isolation and geographic location it could therefore represent an area of outstanding biological value and conservation importance at global scale. Since the 1960s it has been the hideout of armed militias who have been fighting against the government and it has remained largely unexplored till recently. Despite the still scarce information on its biodiversity, the region, based on data collected during the 50s of the last century, has been identified as an important bird area. In 2008 WCS undertook a socioeconomic survey of the communities living around the Kabobo massif which showed that most of the people interviewed were in favour of creating a protected area. More recently, in order to grant access to forest resources to the local communities living along the boundaries of the protected area, seems that the establishment of a National Reserve could be the next step forward (Plumptre et al., 2008; 2015).

4 Kabobo Massif Biodiversity The Kabobo Massif consists of about 1000 km 2 of sub-montane and montane rainforest ranging from about 770 to over 2700 m asl on the escarpment to the west of Lake Tanganyika. Because there is very little forest left within this altitudinal range in Africa, Kabobo is particularly valuable for the conservation of chimpanzees and other forest species occurring at medium elevation (Plumptre et al., 2008). Till the end of the last century, the last survey on vertebrates in the region was conducted by Prigogine in the 1950 s (Prigogine 1960), who documented the bird community and described one species new to science, the Kabobo Apalis (Apalis kaboboensis). As for mammals, the only information available were those of Prigogine s list (1960) of large mammals and short notes on a single squirrel and single colobus monkey both collected by Prigogine (Kerbis et al., 2013) now known as Prigogine's black and white colobus monkey (Colobus angolensis prigoginei). The region is also known to be important for its populations Bongo, possibly a different subspecies, the Red Colobus (Pilocolobus oustaleti foai) which is also a subspecies confined to this region and several other large mammals, including Elephants (Loxodonta africana), Chimpanzee (Pan troglodytes), Lion (Panthera leo), Hippopotamus (Hippopotamus amphibius) as well as other large mammals such as Buffalo (Syncerus caffer) and Giant Forest Hog (Hylochoerus meinertzhageni) (Plumptre et al., 2008; 2015). Civil war and insecurity from 1960 until 2007 prevented further work or its protection. More recently scientists from WCS have discovered four small mammal species, including two bats, a bird and three plants, including a new species of wild ginger, called Aframomum ngamikkense, confined to the higher altitudes of the forest between 1, meters (Kerbis et al., 2013; Fisher et al., 2017). To date, a total of 562 terrestrial vertebrate species and 1413 plant species have been documented for the Kabobo Massif and the neighbouring Luama landscape.

5 Fig. 1 Some of the typical habitat of the Kabobo massif Current conservation challenges After the initial aerial, biological and socioeconomic survey by WCS and partners, was clear that most people wanted to conserve the forest and wanted to create a National Park, that would be connected to the Luama Katanga hunting reserve and could have resulted in a about 4000 km 2 of natural habitat, with at least 1,500 chimpanzees as well as the only habitat of the endemic species of the Kabobo massif forest. In 2015, DRC government decentralization resulted in the creation of multiple new provinces, and the area originally proposed as National Park was suddenly contained within two provinces. Because of this division, local leaders and communities opted to establish the section of the proposed park in South Kivu Province as the M Mbondo Reserve. On August 3, 2016, the Governor of South Kivu designated this protected area, whose name was changed to the Ngandja Natural Reserve to reflect the local culture and heritage of the region. On December 21, 2016, the second section of the proposed park located in Tanganyika Province was declared as Kabobo Natural Reserve, completing the priority protected area complex. Now the various protected areas are manged separately and their combination into a larger, comprehensive, protected area would facilitate and make more efficient its management. Current main threats, at selected sites, especially close to the villages by the lake shore or on the side facing the Luama landscape, are Illegal logging, livestock encroachment, and bush meat hunting.

6 On site DNA sequencing The main goal of this laboratory project was to perform an on site species identification of the some of the samples collected through barcode sequencing technique. Technology A standard genetic species identification workflow includes a DNA extraction step, a DNA amplification step and finally a DNA sequence step. Normally, this procedure take place in a laboratory equipped with at least 9 different lab apparatus and a standard electrical supply. The lab procedure could last almost a week. In order to perform a genetic analysis directly in the field, we have developed a system that could be easily transported, that can be powered by portable batteries and that reduces the time of analysis up to a day. All the laboratory equipment and the reagents has been selected and adapted to be utilized under tropical conditions. The sequencing part has only been possible thanks to MinION portable sequencer. This small sequencer has been developed by Oxford Nanopore Technologies. Our team has established a close collaboration with Nanopore team in order to set up the proper functioning of this portable sequencer out of the lab and in tropical, remote, environment. Preliminary tests Before the expedition we set up an experimental workflow in our standard lab to test all the system and its improvements. We analyzed four different taxa: one mammal, one bird, one amphibian and one reptile. We also performed the analysis starting with different types of biological samples in order to reproduce as much as possible the expeditions conditions. We obtained promising results and the system was ready to be tested again in a field expedition. Kabobo Massif test During the first week of expedition we have successfully sequenced 2 mammal, 2 reptile and 3 amphibian species. The most interesting, preliminary, results of the on-site sequencing, is the one obtained from the tree Pangolin in the genus Phataginus which resulted in having 3% level of divergence from the nearest sequence of P. tricuspis, the species know to occur across the forested areas in Eastern DRC, available on GenBank. In the case of the antelope s sequence, the comparison between sequences obtained in the GenBank database indicates a 96% analogy with Cephalophus callipygus, a species found only in western Africa, in an area 2000 km away from Kabobo. The sister species, as determined from a recent phylogenetic reconstruction, is C. weynsi, whose sequence is missing from the GenBank and could represent the sequenced species. We have also sequenced a frog in the Amietia genus, which resulted putatively new to science, after comparing its DNA sequence with the largest available Amietia sequences database (Eli Greenbaum pers com). In other cases, the on-site sequencing confirmed species identification, as in the case of the lizard Trachylepis maculilabris and the Albertine Rift endemic tree frog Leptopelis mtoewaate.

7 Conclusion The preliminary results show how important is to perform a genetic species identification analysis to quantify the value of the biodiversity present in a certain area. To perform such analysis, it is mandatory to set up a completely portable lab that can work under different field conditions. Our team has developed such us system although it still need some improvements to work with a perfect regime. The use of a portable lab could significantly accelerate the evaluation processes of the biological value of an area. In fact, after almost an year, we are still waiting for the necessary permits, both from DRC and Italian authorities, in order to export the samples collected during the survey and to be able to perform regular DNA analysis under regular laboratory conditions. Results obtained in the field are therefore the only available genetic information on Kabobo biodiversity.

8 Mammals Data on mammal species were collected through the use of camera traps and by interviews with local hunters and assessment of skins and parts of hunted animals. Recorded species confirm many of the species listed by Plumptre et al, 2008 and Kerbis et al., The rare Prigogine's black and white colobus (Colobus angolensis prigoginei) was observed and photographed once (see Fig. 2), at lower elevation. The species is only known for Kabobo and only one sight record of this taxon was made during the recent WCS surveys. An additional specimen of the recently described Myosorex kabogoensis was collected at high altitude (see Fig. 2). The skin of a still unidentified four-toed mongoose (genus Bdeogale) was obtained from a local miner at approximately 2600 m asl (see Fig. 3). Several more mammal samples are awaited to be exported and further analysed. Fig. 2 Some of the mammal species observed on Kabobo massif: from left to right and up to down: L hoest monkey (Cercopithecus lhoesti); the endemic Prigogine's black and white colobus monkey (Colobus angolensis prigoginei); Phataginus cf. tricuspis and, possibly, the recently described and endemic Myosorex kabogoensis (detailed analysis of the specimen is needed to get to a final identification).

9 Fig. 3 Skin of a Bdeogale sp. a still unidentified four-toed mongoose from high elevation.

10 Large mammals checklist Procaviidae Dendrohyrax cf. dorsalis (Fraser, 1855) Elephantidae Loxodonta africana (Blumenbach, 1797) Cercopithecidae Cercopithecus ascanius (Audebert, 1799) Cercopithecus denti (Thomas, 1907) Cercopithecus lhoesti (P. Sclater, 1899) Cercopithecus doggetti (Pocock, 1907) Lophocebus albigena (Gray, 1850) Papio anubis (Lesson, 1827) Colobus angolensis prigoginei (Verheyen, 1960) Pliocolobus foai oustaleti (Trouessart, 1906) Hominidae Pan troglodytes (Blumenbach, 1775) Manidae Smutsia gigantea (Illiger, 1815) Manis cf. tricuspis (Rafinesque, 1821) Felidae Leptailurus serval (Schreber, 1776) Profelis aurata (Temminck, 1857) Viverridae Civettictis civetta (Schreber, 1776) Genetta genetta (Linnaeus, 1758) Genetta maculata (Gray, 1830) Nandinidae Nandinia binotata (Gray, 1830) Herpestidae Herpestes sanguineus (Rüppell, 1835) Mungos mungo (Gmelin, 1788) Bdeogale sp. Hyaenidae Crocuta crocuta (Erxleben, 1777) Canidae Canis adustus (Sundevall, 1847) Mustelidae Mellivora capensis (Schreber, 1776) Suidae Hylochoerus meinertzhageni (Thomas, 1904) Potamochoerus larvatus (F. Cuvier, 1822) Bovidae Oreotragus oreotragus (Zimmermann, 1783) Syncerus caffer (Sparrman, 1779) Tragelaphus euryceros (Ogilby, 1837) Tragelaphus scriptus ornatus (Pocock, 1900) Cephalophus nigrifrons kivuensis (Lönnberg, 1919) Cephalophus weynsi (Thomas, 1901) Philantomba monticola (Thunberg, 1789) Sciuridae Funisciurus carruthersi chrysippus (Thomas 1923) Funisciurus pyrropus akka (de Winton, 1895) Heliosciurus rufobrachium ssp. (Waterhouse, 1842 Heliosciurus ruwenzorii vulcanius (Thomas, 1909) Paraxerus boehmi vulcanorum (Thomas, 1918) Protoxerus stangeri kabobo (Verheyen, 1959 Hystricidae Atherurus africanus (Gray, 1842) Small mammals (non bats) checklist Soricidae Crocidura cf. dolichura Crocidura fuscomurina Crocidura olivieri Crocidura lwiroensis Myosorex kabogoensis Gliridae Graphiurus murinus Muridae Lophuromys aquilus Hybomys aff. univittatus Hylomyscus stella Mus minutoides Mus triton Pelomys minor Praomys degraaffi Praomys jacksoni Praomys sp. Rattus rattus List modified from Kerbis et al., 2013

11 Amphibians and Reptiles The first available information on Kabobo massif herpetological fauna were those collected by the Belgian herpetologist Laurent during the 1950s. The information he published on some of the species collected in Kabobo and his collection, currently housed at the Africa Museum in Tervuren, represented the only available information on Kabobo herpetofauna till few years ago. In 2007 and again in 2012 WCS carried out expeditions to survey the massif, which increased the number of species documented for the region (Plumptre et al., 2008). The following updated amphibian and reptile lists, are the results of both additional analysis, genetic and morphological, on the material collected in Kabobo during 2007 and 2012 expeditions and the findings, although preliminary, of the material collected during the 2017 expedition. To date 25 species amphibians and 29 species of reptiles are known to occur on Kabobo massif of which 5 and 1 respectively, could represent species new to science. Additionally, species like Cardioglossa cyaneospila and Callixalus pictus are poorly known species, known to occur in very few localities. Fig. 3 Some of the Amphibians and Reptiles observed on Kabobo massif: from left to right and up to down: Cardioglossa cyaneospila; Leptopelis mtoewaate, Trioceros schoutedeni and Rhampholeon sp.

12 Amphibians checklist Species Arthroleptidae Arthroleptis sp. 1 Arthroleptis sp. 2 Arthroleptis pyrrhoscelis Laurent, 1952 Cardioglossa cyaneospila Laurent, 1950 Leptopelis cf. kivuensis Ahl, 1929 Leptopelis mtoewaate Portillo & Greenbaum, 2014 Leptopelis sp. Hemisotidae Hemisus cf. guineensis Cope, 1865 Hemisus guttatus (Rapp, 1842) Hyperoliidae Afrixalus sp. Afrixalus wittei Laurent, 1944 Callixalus pictus Laurent, 1950 Hyperolius marginatus Peters, 1854 Hyperolius constellatus Laurent, 1951 Hyperolius sp. 1 Hyperolius sp. 2 Hyperolius cf. langi Noble, 1924 Phrynobatrachidae Phrynobatrachus dalcqi Laurent, 1952 Phrynobatrachus petropedetoides Ahl, 1924 Pipidae Notes putative new to science putative new to science range extension putative new to science putative new to science putative new to science Possibly a range extension Xenopus wittei Xenopus sp. Ptychadenidae Ptychadena uzungwensis (Loveridge, 1932) Pyxicephalidae Amietia cf. nutti (Boulenger, 1896) Amietia ruwenzorica (Laurent, 1972) Amietia sp. putative new to science

13 Phylogenetic reconstruction of Arthroleptis species including samples collected on Kabobo massif. Based on these preliminary findings samples from Kabobo appear to be particularly distinct, as they do not cluster with samples from other localities and have a genetic divergence suggesting that they could belong to species not yet described. More analyses are needed to assess that taxonomic status of the samples (Modified from Hügli, 2013).

14 Phylogenetic reconstruction of Afrixalus species including samples collected on Kabobo massif. As in the genus Arthroleptis and based on these preliminary findings samples from Kabobo appear to be particularly distinct, as they do not cluster with samples from other localities and have a genetic divergence suggesting that they could belong to species not yet described. More analyses are needed to assess that taxonomic status of the samples (Modified from Hügli, 2013).

15 Phylogenetic reconstruction of Phrynobatrachus species including samples collected on Kabobo massif. Based on these preliminary findings samples from Kabobo appear to be particularly distinct from other localities in the Albertine Rift forests and have a genetic divergence suggesting that they could belong to species not yet described. More analyses are needed to assess that taxonomic status of the samples (Modified from Hügli, 2013).

16 Reptiles checklist Species Testudinidae Notes Kinixys belliana Gray, 1831 Agamidae Acanthocercus cyanogaster (Ruppel, 1835) Chamaeleonidae Chamaeleo dilepis Loveridge, 1942 Chamaeleo gracilis Hallowell, 1844 Rhampholeon sp. putative new to science Trioceros johnstoni Boulenger, 1901 Trioceros schoutedeni Laurent, 1952 Gekkonidae Hemidactylus mabouia Moreau De Jonnes 1818 Lygodactylus angularis Günther, 1893 Scincidae Trachylepis maculilabris (Gray, 1845) Varanidae Varanus niloticus (Linnaeus, 1766) Pythonidae Python sebae (Gmelin, 1789) Colubridae Crotaphopeltis hotamboeia (Laurenti, 1768) Dasypeltis atra Sternfeld, 1912 Dasypeltis scabra (Linnaeus, 1758) Dipsadoboa shrevei (Loveridge, 1932) Dipsadoboa unicolor Gunther, 1858 Philothamnus ruandae Loveridge, 1951 Philothamnus heterolepidotus (Gunther, 1863) Philothamnus irregularis (Leach, 1810) Philothamnus semivariegatus (Smith, 1840) Thelotornis kirtlandii (Hallowell, 1844) Lamprophiidae Boaedon lineatus Dumeril, Bibron & Dumeril 1854 Duberria lutrix (Linnaeus, 1758) Lycophidion capense (Smith, 1813) Lycodonomorphus bicolor (Gunther, 1893) Elapidae Naja annulata stormsi Peters, 1876 Naja melanoleuca Hallowell, 1857 Viperidae Atheris nitschei Tornier, 1902

17 References Fischer E, et al (2017) A new species of Aframomum (Zingiberaceae) from D.R. Congo. Phytotaxa. 298 (3): Hügli D.M. (2013) Is molecular barcoding always best? Amphibian diversity and biogeography in Nyungwe National Park, Rwanda. Unpublished Master Thesis in Geosciences. Department of Environmental Sciences Biogeography. University of Basel Kerbis, Julian & H. Huhndorf, Michael & Plumptre, Andrew & Hutterer, Rainer & Kaleme, Prince & Ndara, Benjamin. (2013). Mammals, other than bats, from the Misotshi-Kabogo highlands (eastern Democratic Republic of Congo), with the description of two new species (Mammalia: Soricidae). Bonn Zoological Bulletin Plumptre, A.J., Kujirakwinja, D., Matunguru, J., Kahindo, C., Kaleme, P, Marks, B., and Huhndorf M Biodiversity surveys in the Misotshi-Kabogo and Marungu regions of eastern Democratic Republic of Congo with a focus on chimpanzees. Albertine Rift Technical Reports No. 5 Plumptre, A.J., Kujirakwinja, D., Bamba, A., Kibambe, J.P., Muhamba, C. and Ilambu, O. (2015) Conservation Action Plan for the Kabobo-Luama Landscape. Unpublished report to Rainforest Trust, CEPF and USFWS. Prigogine A (1960) La faune ornithologique du Massif du Mont Kabobo. Annales de Musée Royal du Congo Belge, series 8, 85: 1 47