P1-33BEL-000177 – Final Report

FINAL REPORT NAMIBIA BBI SSFA April 2020

Development of Institutional Synergy to Enable Inventorying and Cultivation of Priority Species for Conservation in the Southern Africa Region: Medicinal in the Family

The overall objective of this Project is to foster long-term cooperation between key and relevant Institutions in Southern Africa on addressing the unsustainable exploitation of medicinal plants in the Region. The initial focus is a group of plants in the Apocynaceae family that recently became popular in northern Namibia. The project endeavours to i) Conduct traditional and molecular-based taxonomic work to clarify the identity and phylogeny of the targeted species, ii) Contribute to an existing public DNA sequence database (e.g. NCBI), iii) Establish propagation initiatives to reduce pressure on the wild resource, and iv) Develop policy to accord the targeted species legal protection in Namibia and other range states. i) and ii) constitute the immediate Project objectives to which this CBD Bio-Bridge Initiative Small-Scale Funding contributes, through six key activities. All six activities were successfully implemented over the period March 2019 – March 2020, as detailed in the following report:

1. Outline in detail the activities implemented during the Project.

Activity 1: Collection trip to northern Namibia – Herbarium voucher specimens of the targeted plants and leaf samples for molecular analysis will be collected from various sites in northern Namibia.

Feedback: Herbarium specimens, leaf samples as well as soil samples were collected from 85 sites across nine Namibian regions in March – April 2019 (see Collection sites 2019 in “Report_BBI Namibia”). However, 2019 suffered a severe drought that affected specimen collection, especially in the arid northwest. Consequently, fieldwork was shortened by two weeks as the area west of Sesfontein/Opuwo was deemed too dry to be worth visiting. This year, 2020, has received good rains, presenting an opportunity to visit the area that was left out.

Activity 2: Visit to Herbarium of Lubango (LUBA) – Namibian partners will visit LUBA to familiarise themselves with the facility and its staff, identify how it can enhance the current Project, discuss the logistics of the upcoming trips to the African Centre for DNA Barcoding (Activity 3) and laboratory training workshop in Windhoek (Activity 4), and determine the feasibility for a joint field collection in Angola in a subsequent phase of the project.

Feedback: Three Namibian botanists visited LUBA and the host Institution ISCED for two days in September 2019. They familiarised themselves with the dhingila specimens held in the Herbarium and other taxa of interest, networked with staff, met the Director of ISCED, Directors of ISPT, and undertook a botanical excursion around Lubango. They were informed that LUBA was in the process of being transferred to the management of the ISPT. The Herbarium recommended a student from ISPT 1

P1-33BEL-000177 – Final Report and one from the University of Agostinho Neto in Luanda to attend the laboratory training workshop in Windhoek.

The networking opportunity that the trip provided will go a long way in supporting continuation of the dhingila work and in contributing to other prospective projects of common interest. Angola is one of Namibia’s major biodiversity stakeholders; the two countries currently share two Trans-frontier Conservation Areas, SIONA and KAZA.

News article: http://www.pcu.uct.ac.za/botanists-visit-lubango-angola-092019

Activity 3: Visit to African Centre for DNA Barcoding (ACDB) – Namibian and Angolan partners will visit the African Centre for DNA Barcoding in Johannesburg, South Africa, to familiarise themselves with the facility, identify how the sequencing services required from the Centre (Activity 5) can be accelerated, due to time constraints, and to strengthen linkages to support potential biodiversity projects in the visiting countries in accessing the facility in future.

Feedback: Four Namibian botanists and two Angolan students undertook a one day visit to the ACDB in December 2019. The delegates were treated to presentations on various projects of the ACDB, a demonstration of the LifeScanner, introduction to the Lab-in-a-box, and a tour of the ACDB laboratory facility. Since Activity 4 preceded the visit the delegates took the samples to ACDB for sequencing. The visit provided insight into the Centre and a platform for communicating services needed or potential opportunities for collaboration.

Activity 4: Molecular analysis laboratory training workshop (extraction and amplification) – A training workshop will be held at the University of Namibia to train Namibian and Angolan participants in laboratory methods for DNA extraction and amplification, using leaf samples collected in Activity 1. Emphasis will be placed on DNA barcoding, but other molecular analysis techniques will also be considered if deemed feasible.

Feedback: A total of eight Namibian and two Angolan participants took part in the laboratory training workshop, which was held towards the end of 2019. They extracted and amplified DNA barcodes from 102 leaf samples of mostly dhingila species collected in the field, but some other Apocynaceae as well as other species of interest field-collected or taken from the National Herbarium or Botanic Garden were also included. The main training session took place from 25 November to 4 December 2019 and was blessed with opening remarks by the Head of the Biological Sciences Department, Prof. John Mufune. The main training was preceded by a trial run which processed the first 50 samples and tested three plant barcode markers, recommending matK for the main training session. News article: http://www.pcu.uct.ac.za/barcoding-dhingila-namibia

Activity 5: Sequencing – Samples from Activity 4 will be sent to the African Centre for DNA Barcoding (ACDB) in South Africa for sequencing.

Feedback: 48 samples were taken to the ACDB on 6 December 2019 for sequencing. Results were received on 6 March 2020.

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Activity 6: Informatics – This activity will entail analysis of the molecular data matrix and possibly morphometric analysis.

Feedback: Towards the end of the training workshop in December participants were introduced to sequence editing and construction of phylogenetic trees using sequences of the Namib Desert endemic Moringa ovalifolia. Upon receiving dhingila sequences from the ACDB, 10 of them were selected on the basis of geographic representativeness, and were then edited and submitted to the Barcode of Life Database (BOLD). The document “From Field to BOLD” in the folder “Report_BBI Namibia” outlines the workflow.

Comparison of collected herbarium specimens with material in the National Herbarium of Namibia and blasting of chloroplast MatK DNA barcode sequences obtained from the training workshop against the Genbank (NCBI) sequence database have identified the targeted dhingila species as macrantha (Clotzsch) Schltr. (Apocynaceae) and sylvestre (Retz.) R.Br. ex Schult (Marsdenia sylvestris (Retz.) P.I. Forst.) (Apocynaceae). Both species are used inseparably as “they taste the same, smell the same, and are both effective”, according to the words of a local dhingila trader who identified them as the “female dhingila” (M. macrantha) – based on the broader-shaped fruit and as “male dhingila” (G. sylvestre) – which has a slender fruit. It also appears that a vine Fockea species (Apocynaceae), a wild jasmine (Oleaceae), and possibly also the alien invasive rubber vine (Apocynaceae) are sometimes confused with the Marsdenia and Gymnema species, especially in the absence of fruit and flowers. While confamilials may present similar phytochemicals and curative properties there is clearly a need for awareness work to ensure that the local people can distinguish the “real dhingila” species.

The literature indicates that M. macrantha naturally occurs in a number of African countries, while G. sylvestre is more widespread in tropical to subtropical Africa and beyond, with natural distribution extending as far east as Arabia, India, and China (Bruyns 2014). The leaves of G. sylvestre, marketed as Indian Ayurveda herbal tea, are used around the world mainly for slimming and to treat diabetes (see e.g. ScienceDirect and this site). While in Namibia the main focus is on the root, some of the ailments targeted are similar (see the document “Dhingila Products” in “Report_BBI Namibia”). There is emerging interest in Namibia to protect dhingila as an indigenous biological resource with potential health and commercial value as well as to ensure benefit-sharing from commercialisation of the traditional knowledge (TK) and any intellectual property associated with the plant, in line with the Nagoya Protocol and the Namibian Access to Biological and Genetic Resources and Associated Traditional Knowledge Act. However, claims of ownership of the G. sylvestre resource and TK could compete with those that could emerge from other range states such as India, where there is a documented long history of traditional use and an established trade. Evidence that there is variation significant enough to warrant delimitation of the Namibian and other regions’ G. sylvestre as separate taxa at any taxonomic level will help avert potential conflict. A degree of evolutionary divergence across such a wide geographic and climatic range is expected. Northwestern Namibia could possibly represent one of the driest, if not the driest, and westernmost habitats of G. sylvestre, which presents an opportunity for comparison with specimens from other regions. Taxonomic revision of the initial description has seen subsequent placement of the Namibian species under Marsdenia sylvestris 3

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(Retz.) P.I. Forst. and (Retz.) R.Br. ex Schult. (see photo of herbarium specimen “Marsdenia sylvestris-Gymnema sylvestre” in ‘Report_BBI Namibia’). Further phylogenetic scrutiny will contribute to clarifying the evolutionary relationship of the Namibian species with Gymnema and Marsdenia species and to establishing which taxonomic name would be most appropriate. Preliminary phylogenetic analysis will be conducted using the barcode nucleotide sequences generated in the present Project and those from other regions mined from the Genbank. Results will be presented through a poster presentation at the upcoming 22nd AETFAT Congress in September 2020 in Zambia.

Field observations have indicated that both dhingila species are widespread throughout northern Namibia and locally abundant in some areas, while trade and utilisation are still relatively small-scale and limited to traditional personal use in the northcentral part of the country. Although some local people reported incidences of plant theft and depletion in their respective areas the problem is probably localised, while the prolonged drought in 2019 likely also affected dhingila regeneration. It is, therefore, unlikely that the current scale of wild harvesting is unsustainable. However, this could change quickly if demand increases and local suppliers start exporting dhingila products – especially as dried, raw plant material. A local trader is actively looking for such an opportunity, and while this would be a welcomed contribution to livelihoods a strategy needs to be in place to manage trade sustainability and potential conservation impacts.

2. What are the main outcomes of the Project? How these results impact the state of the biodiversity? How this Project generated social and economic benefits? Please annex any written relevant document.

The Project has i. promoted molecular techniques and application to compliment traditional taxonomic methods for identification to protect priority species in Namibia and the region, ii. has contributed to the enhancement of technical collaboration across institutions in the region to capitalize on expertise and facilities available, and iii. has contributed to sustainable use of indigenous botanical resources for utilization in traditional medicine in Namibia and southern Africa.

3. Describe how the Project enhanced the Technical and Scientific Cooperation (TSC) between the Parties and organizations involved in it.

Through all activities of the Project Angolan, Namibian and South African botanists, students as well as laboratories and Technologists had the opportunity to interact, network and exchange skills that contributed to the achievements of the Project, and will continue to benefit individual or collaborative projects in the future.

4. Which activity or approach efficiently succeeded to foster sustained TSC and why? Which activities or approaches would be done differently, now that you have experienced them, and why?

All six activities contributed significantly to TSC. This was possible because they were focused and relevant to the main objective of the Project. What could perhaps be done differently in Activity 4 would be to order and purchase lab materials through UNAM rather than the Project doing this directly

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P1-33BEL-000177 – Final Report and paying more. As a regular customer and a large one UNAM has discount privileges and procurement arrangements with some of the suppliers in the country and the Region.

5. Describe the in-kind contribution of the different stakeholders involved in the Project and how this type of contribution could be increased in future projects.

The Directorate of Forestry and its NBRI Subdivision provided, with much appreciation, a field vehicle and fuel for the fieldwork undertaken in March - April 2019. UNAM availed lab space and a competent Technologist. The Herbarium of Lubango ensured that the Namibian botanists made the most of the short visit and organised meetings with Institution Directors. The ACDB put together a programme with presentations and activities and provided lunch. They may have also undertaken some DNA recovery steps at no extra cost as they had indicated that they would do so if need be, because the dhingila samples taken to the ACDB were not on ice during transit and it is possible that there was some DNA degradation. To encourage continued support and to help maintain the good relationship established it is important to keep communicating and updating collaborating institutions and staff on developments and any publications stemming from the Project.

6. If the Project included a participation of major groups like business, subnational and local authorities, NGOs, youth, women, indigenous peoples and local communities, what would be your advice to engage efficiently this specific group?

Our advice through experience as government employees would be to ensure that there is clear and unambiguous communication with such groups to avoid creating expectations that cannot be met. There is often an unrealistic expectation on government to assist in kind or financially, especially with regard to community initiatives.

7. Any other lessons learned to share with the Bio-Bridge team?

We have further learnt that to minimise delay such as the signing of the Project Agreement all the officials who would be signing off on the document should be briefed collectively and well in advance to ensure that they are all informed and have a common understanding about the Project and what needs to happen to facilitate implementation. It took a while to get this Project’s Agreement signed by the MET, with much of the delay experienced at the recommendation stages before the document got to the Accounting Officer’s desk.

8. How this Project could be pursued to maintain or scale up the positive results obtained? How this Project could be replicated in another region?

Platforms such as the upcoming AETFAT Congress are useful opportunities to show-case the Project and to network with potential collaborators.

9. What would be your vision for future, longer term cooperation between the Parties and organizations involved in the Project? Describe any roadmap that was discussed among relevant stakeholders to achieve this.

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The Project will continue to seek additional funding to further the work started in this first phase and to capitalise on the local and regional network that has been built. The taxonomic work could possibly extend to include other genetic markers and techniques for phylogenetic analysis, while propagation trials would need to be pursued as a strategy to reduce pressure on the wild resource should demand increase. Policy would also need to be developed to accord the species protected status.

10. Please compile and attach links to or copies of any media/ video/ articles on the Project.

The folder “Photo gallery and documents” herewith annexed contains photographs, video clips and documents. “Meet the participants” contains a list of participants.

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Meet the Participants

Ms Kirsti Nghidinwa: Chief Conservation Scientist, Ministry of Environment and Tourism • Project Coordinator, Fieldwork, LUBA, Barcoding Training Workshop, ACDB Ms Esmerialda Strauss: Head of National Botanical Research Institute (NBRI) • Barcoding Training Workshop, ACDB Ms Frances Chase: Curator, National Herbarium of Namibia, NBRI • LUBA, Barcoding Training Workshop, ACDB Mr Leevi Nanyeni: Forester, NBRI • Fieldwork, Barcoding Training Workshop Mr Stephen Carr: Senior Forester, NBRI • Fieldwork Prof. Fernanda Lages: Herbarium of Lubango (LUBA) • Host Mr Augustinus Mbangu: Technologist, University of Namibia • Barcoding Training Workshop Ms Quanita Daniel: Forester, NBRI • Barcoding Training Workshop Ms Kahimbi Sikute: Forester, NBRI • Barcoding Training Workshop Dr Elizabeth Ndeunyema: Lecturer, University of Namibia, Ogongo Campus • Barcoding Training Workshop Ms Bianca Pearce: Student, University of Namibia • Barcoding Training Workshop Dr Timothy Sibanda: Senior Lecturer, University of Namibia • Barcoding Training Workshop Mr Frans Kamenye: Game Products Trust Fund • Grant Manager Prof. Michelle van der Bank: African Centre for DNA Barcoding (ACDB) Director • Host, sequencing services 7

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