2.1.1. Online training

The training of Indonesian participants was initiated in the context of the Center for Botanicals and Chronic Diseases, or CBCD (https://cbcd.rutgers.edu/), funded by the USA National Institutes of Health – Fogarty International Center (NIH–FIC). This centre provides training across a spectrum of research disciplines, spanning basic biomedical and botanical to clinical and applied sciences, including translational and implementation science. Using a culturally sensitive approach, it connects traditional use of botanical therapeutics in participating countries – Indonesia and Tajikistan – with the state-of-the-art research capabilities of participating USA institutions. By combining intensive training in Western science with robust knowledge of traditional botanical medicines, the CBCD program aims to cultivate scientists who can integrate these two systems for the benefit of participating countries. Instruction is organised around four sections: Ethics Core, Botanical Core, Analytical Core and Drug Development Core.

The 2021–22 cohort of the program included twenty-five trainees, sixteen of whom were Indonesian from three Indonesian institutions. Trainee selection considered academic performance, English proficiency, and personal research interests. Training was conducted online using the Rutgers University Canvas learning management system and Zoom video conferencing, avoiding disruption by the global COVID-19 pandemic lockdowns. A mix of synchronous and asynchronous lectures was used for instruction delivery, and regular assignments and examinations were used to monitor progress and assess understanding. In addition, trainees undertook research on a self-selected topic under the guidance of mentors from Indonesia and the USA.

2.1.2. Practical RAMES-STN workshop

After the easing of COVID-19 lockdown measures, an immersive in-person full-day training workshop on RAMES and STN technologies, funded by the Asia-Pacific Network for Global Change Research (APN), took place in July 2022 in the Mt. Halimun-Salak National Park in West Java. This protected, highly biodiverse location was selected to underscore the portable character of these technologies while reinforcing the notion that our natural environment contains a vast array of phytochemicals with invaluable bioactivities. The seventeen Indonesian trainees included six PhD students, eight MS students, and three BSc students. Thirteen of these trainees were selected from the group who had already completed CBCD training, while the remaining four were candidates for the subsequent CBCD cohort.

The large number of workshop participants, as well as their heterogeneous levels of technical expertise, potential language barriers and time limitations, all posed significant potential challenges. These were addressed prior to the event through careful organisation, and all trainees were also sent a detailed manual in English and asked to read it before the workshop. On workshop day, two Rutgers scientists provided instruction in plant collection methods, including documentation and species identification. They subsequently learned the rapid preparation of ethanolic extracts and their immobilisation onto glass microfiber discs using RAMES technology and then learned to apply STN field-adapted bioassays to assess the antibacterial, antifungal, and antioxidant bioactivities of the freshly prepared extracts. All technical details about these methodologies are published in Skubel et al. (2018). After the workshop, participating researchers from Universitas Nasional (UNAS) received a kit containing all materials for performing RAMES extraction and STN functional assays on 1,000 samples.

2.2.1. Collection and extraction of samples using RAMES technology

The development of the pilot Indonesia MAGIC Library was spearheaded by a team of Indonesian scientists from UNAS. This team comprised three senior researchers specialising in medicinal plants, environmental sciences, and ethnobotany, along with three young scientists who had completed the training described in Section 2.1. Rutgers scientists also assisted via regular progress meetings and participation in some collection activities. The UNAS team conducted field collection trips to 25 areas of Indonesia (Figure 1, Table 1), and local guides assisted with preliminary taxonomic identification. Species selected for the library at this pilot stage were Indonesian species with previously known medicinal properties. Whenever a species of interest was preliminary identified in the field, the team took pictures of the whole plant, as well as flowers or fruits if present, and recorded its GPS coordinates prior to collecting it using scissors and placing it inside a labelled plastic bag. The collected samples were then individually photographed using a portable lightbox just before their extraction. These images would later be used to confirm preliminary identification using taxonomic keys.

For each species, samples were collected from various tissues – leaves, stems, rhizomes, tubers, flowers, fruits and/or seeds – depending on availability and with minimal disruption to the plants. Samples were processed using RAMES technology while still in the field to minimise phytochemical degradation during transport. For each species, two grams of plant material were extracted in 5 ml of 95% ethanol using a specially modified Dremel®; cordless rotary tool. The extract was filtered and loaded onto 10 mm borosilicate glass microfibre discs, which were dried using a portable fan (Skubel et al., 2018). The process was repeated until reaching a minimum of 40 discs per sample, each holding 90 µl of extract. Dry discs were stored in resealable plastic bags and transported back to UNAS for storage at −20 °C.

2.2.2. Long-term sample storage and data management

Once dry, discs from each sample were stored in resealable plastic bags, each labelled with the corresponding unique sample ID and species name. These bags were then stored in a −20 °C lockable freezer within the UNAS laboratories.

While the library was being created, comprehensive data about each sample was kept in an Excel spreadsheet. This information included unique identification numbers, scientific and common names, family, plant organ collected, name of collector, GPS coordinates, plant habitat, whether the sample was from a cultivated species, and a summary of its traditional medicinal uses. Upon formally establishing the Indonesia MAGIC Library as a distinct unit within the Center for Medicinal Plants Research (CMPR) at UNAS, a section was created within the CMPR website to centralise all available information.

3.3.1. First international conference and signature of strategic partnership

In June and July 2022, three forums for scientific discussion and networking were held alongside the RAMES-STN workshop described in Section 2.1.1. These included a small conference between UNAS and Rutgers scientists, a formal meeting and discussion between scientists from Rutgers and the Indonesian National Research and Innovation Agency (BRIN) – Indonesia’s government agency in charge of all government affairs in the field of research and technology–, and most notably, a novel International Conference organised by UNAS, the “1^st International Conference on Natural Products and Chronic Diseases 2022”, held in Jakarta, Indonesia (iconference-ncd.unas.ac.id). This hybrid conference, focusing on the impact of plant natural products on human health, featured speakers from five universities across four countries and garnered wide coverage from local newspapers. The list of attendees, which included a total of 206 participants, featured representatives from fifteen Indonesian and foreign institutions (Figure 5, Table 4).

These discussion forums solidified a strategic partnership among UNAS, BRIN, and Rutgers, leading to the signature of a Memorandum of Understanding (MOU) for long-term collaboration. At the end of 2022, a BRIN researcher and former scientific advisor to the Indonesian president conducted research at the Raskin laboratory at Rutgers as a Fulbright scholar, facilitating further discussion and networking opportunities.

3.3.2. Second international conference and second training workshop

These developments culminated in the rapid organisation of a second, larger international conference as well as a second RAMES-STN training workshop, “The International Conference and Workshop in conjunction with the 8th Indonesia Biotechnology Conference 2023” (https://www.icw-ibc2023.com/). The combined event was jointly organised by BRIN, UNAS, the Indonesian Biotechnology Consortium (KBI), Universitas Mahasaraswati Denpasar (UNMAS) and Rutgers. The two-day hybrid conference took place in Denpasar, Indonesia, in May 2023 and explored the intersection of biodiversity, biotechnology, and health for enhancing sustainable development. It featured nineteen keynote speakers from four countries and attracted 293 participants from 61 institutions (Table 5, Figure 6).

The second RAMES-STN training workshop took place at the Bali Botanical Gardens the day after the conference (Figure 7), following the same full-day format and covering the same topics as the workshop already described in Section 2.1.1. The nineteen participants were conference attendees who had registered for the workshop due to their personal interest, and they included ten researchers, five PhD students, three MSc students and one senior high school student, coming from fourteen institutions across Indonesia (from Java, Sumatra, and Sulawesi), as well as from Okinawa in Japan and from the USA (Figure 2, Table 2). Careful workshop planning prior to the event helped the training run smoothly despite the potential challenges posed once again by the diverse backgrounds of participants, the time limitations and potential language and technical barriers. Once again, the workshop was not formally assessed but rather followed by a post-workshop Q&A session and discussion, during which trainees offered highly positive feedback, expressing eagerness to incorporate the learned techniques into their research. Post-workshop, BRIN representatives received a kit containing all necessary materials for performing RAMES extraction and STN bioassays on 1,000 samples. Both events received thorough coverage from local newspapers as well as the BRIN and KBI websites. Furthermore, two additional Indonesian universities have recently joined the Indonesia MAGIC Library and will participate in future sample collections: Universitas Surabaya (UBAYA) from East Java and Universitas Pancasila from Jakarta.

3.4.1. Suggested next steps

As a completely new resource, the Indonesia MAGIC Library provides numerous avenues for contributing to research, conservation, and economic development in Indonesia. A suggested starting point would be conducting quality control and stability studies of the RAMES disc contents. This can be achieved by eluting the extracts, identifying and quantifying metabolites through UPLC/MS analysis, and comparing them with freshly prepared extracts. A subsequent step could involve rapid functional screening by assessing the antioxidant, antifungal, and antibacterial bioactivities of the extracts with the corresponding STN bioassays.

As Rutgers scientists have developed a broad range of simple, rapid, and portable bioassays beyond the antioxidant, antibacterial, and antifungal – including, for example, assays to detect anti-roundworm, anti-flatworm, anti-Leishmania, or wound healing properties – this rapid functional screening can be broadened to include many bioactivities of interest. Upon identifying extracts with promising bioactivities, it is recommended that detailed metabolic profiling and extensive quantitative analysis of their constituents be performed.

The present pilot version of the library focused on Indonesian plant species with known medicinal properties. Future expansion efforts of the library are also planned to include Indonesian endemic and endangered plant species. In parallel, it would be of great interest to expand the collection by including marine organisms, fungi and insects. Another course of action could be resampling already catalogued species in different seasons or under varied stress conditions to conduct comparative studies, which would provide valuable insights into the impact of environmental and biological stresses on metabolomic profiles. In any case, each new RAMES sample collected in the future should also include a supplemental small, dehydrated piece of tissue (under one gram would suffice) within the same envelope for DNA barcoding and other sequencing strategies, which would serve as a means to validate species identification, thus providing a practical substitute for traditional herbarium vouchers. The incorporation of genomic samples into the Indonesia MAGIC Library was an integral part of the initial vision, as indicated by the acronym MAGIC, which stands for Metabolome And Genome Innovation and Conservation. Consequently, Rutgers scientists have developed and validated a rapid method for field collecting plant samples for DNA studies and barcoding, using silica gel as a drying method (Skubel et al., 2018). In the future, it might also be interesting to develop a near-infrared (NIR) spectroscopy method for the metabolite profiling of RAMES samples, which could serve as a simple, portable, and cost-effective alternative to the current LC-MS method.

Lastly, raising awareness of this unique biological resource at academic gatherings like conferences and scholarly forums is crucial. By doing so, we hope to initiate sample sharing among researchers, leading to the establishment of collaborative research projects within Indonesia and worldwide. Moreover, we expect that awareness of the Indonesia MAGIC Library will encourage more institutions to participate in the collection of samples, thus ensuring its growth over time.

3.4.2. Challenges faced and recommendations for the future

It must also be noted that the project encountered several challenges described below. While these were tackled as effectively as possible, they require attention and strategic planning, providing valuable lessons for any future initiatives. These issues also emphasise the need for subsequent studies to validate the quality of the library.