ABSTRACT. The Nee Soon stream drainage in the Central Catchment Nature Reserve is virtually the last remaining fragment of primary freshwater swamp forest in Singapore. The forest type has been poorly studied in the Southeast Asia. The hydrology, water quality, as well as aquatic flora and fauna all have great theoretical and practical significance. The ecology and management of the Nee Soon freshwater swamp forest are reviewed, with remarks on their national, regional and global contexts. This review sets the scene for a three-year integrated conservation and management study completed in 2016.

Food mislabelling is a growing world-wide problem that is increasingly addressed through the authentication of ingredients via techniques like mass spectrometry or DNAsequencing. However, traditional DNA sequencing methods are slow, expensive, and require well-equipped laboratories. We here test whether these problems can be overcome through the use of Nanopore sequencing. We sequenced 92 single and 13 mixed-species samples bought in supermarkets and restaurants in Singapore which has a large and diverse seafood trade. We successfully obtained DNA barcodes for 94% and 100% of the single-and mixed-species products after correcting the numerous sequencing errors of MinION reads with a correction pipeline optimized for DNA barcodes. We find comparatively low levels of clear-cut mislabelling for single-species samples (7.6 %) while the rates are higher for mixed-species samples (38.5 %). These low rates are somewhat deceptive, however, because of the widespread use of vague common species names that do not allow for a precise assessment of the expected ingredients. With regard to the clearly mislabelled single-species products, higher-value products (e.g., prawn roe, wild-caught Atlantic salmon, halibut) are replaced with lowervalue ingredients (e.g., fish roe, Pacific salmon, arrowtooth flounder) while more serious problems are observed for mixed-species samples. Cuttlefish and prawn balls repeatedly contained pig DNA and 100% of all mixed samples labelled as containing crustaceans ('crab', 'prawn', 'lobster') only yielded fish barcodes. We conclude that there is a need for more regular testing of seafood samples and suggest that due to speed and low-cost, MinION would be a good instrument for this purpose. We also emphasize the need for developing clearer labelling guidelines..

10Food mislabelling is a growing world-wide problem that is increasingly addressed 11 through the authentication of ingredients via techniques like mass spectrometry or DNA-12 sequencing. However, traditional DNA sequencing methods are slow, expensive, and 13 require well-equipped laboratories. We here test whether these problems can be 14 overcome through the use of Nanopore sequencing. We sequenced 92 single and 13 15 mixed-species samples bought in supermarkets and restaurants in Singapore which has a 16 large and diverse seafood trade. We successfully obtained DNA barcodes for 94% and 17 100% of the single-and mixed-species products after correcting the numerous 18 sequencing errors of MinION reads with a correction pipeline optimized for DNA 19 barcodes. We find comparatively low levels of clear-cut mislabelling for single-species 20 samples (7.6 %) while the rates are higher for mixed-species samples (38.5 %). These low 21 rates are somewhat deceptive, however, because of the widespread use of vague 22 common species names that do not allow for a precise assessment of the expected 23 ingredients. With regard to the clearly mislabelled single-species products, higher-value 24 products (e.g., prawn roe, wild-caught Atlantic salmon, halibut) are replaced with lower-25 value ingredients (e.g., fish roe, Pacific salmon, arrowtooth flounder) while more serious 26 problems are observed for mixed-species samples. Cuttlefish and prawn balls repeatedly 27 contained pig DNA and 100% of all mixed samples labelled as containing crustaceans 28 ('crab', 'prawn', 'lobster') only yielded fish barcodes. We conclude that there is a need 29 for more regular testing of seafood samples and suggest that due to speed and low-cost, 30 MinION would be a good instrument for this purpose. We also emphasize the need for 31 developing clearer labelling guidelines. 32 33 34

Many invertebrate and plant species are difficult to identify even by taxonomic experts. This has created a major obstacle for understanding the ecology of tropical environments. Here we explore the use of new large-scale, cost-effective approaches to species identification using Next-Generation Sequencing ("DNA barcodes"). Due to the rapid drop in sequencing cost, such barcodes have the potential to help with many identification tasks and they will facilitate regular monitoring of habitats. We use this approach to explore the species diversity of Nee Soon freshwater swamp forest and provide taxonomic identification tools for the fauna and flora of the forest. DNA-barcode libraries were generated for the flora (>1000 barcodes; 170 chloroplast genomes) and fauna (ca. 3000 barcodes). In addition, highresolution images of 502 animal and 200 plant species were placed on an online image database ("Biodiversity of Singapore"). These images are available to help experts and non-experts alike to identify and appreciate these species. The new databases document Nee Soon's impressive diversity, but they are also important for in-depth studies of fauna-floral species interactions. For example, the plant barcodes were used to reconstruct the diet of Raffles' banded langur based on faecal samples. Overall, we find that the fauna in Nee Soon freshwater swamp forest is very diverse and includes many rare species, and that the species composition is very distinct from those living in surrounding habitats. Animal specimens are readily sequenced, while plant specimens (especially those represented by sapwood samples) remain a challenge. However, newer techniques (e.g. based on genome skimming) are starting to help with obtaining plant DNA-barcodes.

The Nee Soon freshwater swamp forest is a vital area for biodiversity conservation in Singapore. A survey of the aquatic macroinvertebrates in the streams of the Nee Soon drainage was carried out to capture a representative sample of the communities present. Here, we present the different groups of macroinvertebrates sampled as well as their abundance and distribution within the freshwater swamp forest. An annotated checklist of the orders of the macroinvertebrates found in the freshwater swamp forest follows, together with information on their distribution and abundance within the Nee Soon catchment.