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The approach of combining cost-effective nanopore sequencing and emerging environmental DNA (eDNA) metabarcoding could prove to be a promising tool for biodiversity documentation, especially in Malaysia. Given the substantial funding constraints in recent years, especially in relation to the country's biodiversity, many researchers have been limited to conduct restricted research without extended monitoring periods, potentially hindering comprehensive surveys and could compromise the conservation efforts. Therefore, the present study aimed to evaluate the application of eDNA metabarcoding on freshwater fish using short reads generated through nanopore sequencing. This assessment focused on species detection in three selected rivers within the Endau Rompin Landscape in Malaysia. Additionally, the study compared levels of species detection between eDNA metabarcoding and conventional sampling methods, examined the effectiveness of primer choice, and applied both metabarcoding and shotgun sequencing to the eDNA approach. We successfully identified a total of 22 and 71 species with an identification threshold of >97% and >90%, respectively, through the MinION platform. The eDNA metabarcoding approach detected over 13% more freshwater fish species than when the conventional method was used. Notably, the distinction in freshwater fish detection between eDNA primers for 12S rRNA and cytochrome oxidase I was insignificant. The cost for eDNA metabarcoding proved to be more effective compared to conventional sampling with cost reduction at 33.4%. With favourable cost-effectiveness and increased species detection, eDNA metabarcoding could complement existing methods, enhance holistic diversity documentation for targeted habitats and facilitate effective conservation planning.

Environmental DNA (eDNA) metabarcoding is a promising tool for regular biological monitoring, especially for freshwater fish, which are facing tremendous threats worldwide. The application of eDNA detection is a dramatic improvement on common methods of biomonitoring as it produces tangible results in a short time with low effort and little expense. However, the accuracy of the technique is largely dependent on the availability of genetic references for the target organisms. In this study, we investigated the availability of genetic resources for freshwater fish in Malaysia in three public depositories, National Center for Biotechnology Information, Barcode of Life Data System, and Mitochondrial Genome Database of Fish, focusing on seven targeted genes of mitochondrial DNA. We found that only 68.6% of freshwater fish found in Malaysia had information on at least one of the seven targeted genes, with data on cytochrome c oxidase subunit 1 being most commonly available. Genetic information for threatened and endemic species were underrepresented (33.3%-41.7%), yet fish of commercial value and invasive species were well explored genetically. Although there is still room for improvement to achieve comprehensive and reliable genetic resource information for freshwater fish in Malaysia, the application of eDNA metabarcoding is still highly relevant. This is since the current decline in freshwater fish diversity in Malaysia is alarming and because the technique will assist in the ongoing effort to generate new genetic references for Malaysian freshwater fish.

Malaysia is blessed with lush tropical rainforests that harbour an exceptional diversity of amphibians and reptiles. However, compared to other animal groups, amphibians and reptiles have received limited attention in research, despite their ecological significance. With amphibians and reptile species having been declining rapidly due to anthropogenic activities, there is a pressing need to conserve these species and their habitats. Environmentally Sensitive Areas (ESAs) are designated regions that are beneficial due to their critical role in providing essential ecosystem services and serving as repositories of biodiversity. Nonetheless, the classification of ESAs in Malaysia lacks biological elements and only focuses on physical attributes. To enhance the current ESA classification framework by integrating biological components, there is an urgent need to obtain information on diversity and habitat in Malaysia. Therefore, the objectives of this study were twofold: to determine the diversity of amphibians and reptiles in Gunung Belumut Amenity Forest and to conduct a comparative analysis between the herpetofauna assemblages in Gunung Belumut with other forest reserves in Peninsular Malaysia. The survey was carried out between March and June 2022, with additional sampling conducted in February 2023. The Visual Encounter Survey (VES) and pitfall trap methods were employed to survey the herpetofauna species, focusing on both aquatic and terrestrial habitats within the study area. A total of 210 individuals representing 38 species of herpetofauna were recorded, comprising 18 amphibian and 20 reptile species. Amongst the observed species, Limnonectes blythii was the most frequently encountered amphibian, with 59 individuals observed, while the dominant reptile species was Cyrtodactylus consobrinus, represented by eight individuals. This pioneering study serves as a vital baseline documentation of the amphibian and reptile assemblages in Gunung Belumut Amenity Forest. It

Gut microbiome studies have gained significant attention in recent years due to their potential in unveiling the role of microbial communities in animals’ health and ecological processes. However, the lack of standardized protocols in sample handling and processing across studies introduces variability, impeding the comparability of findings. This study addresses this issue by examining methodological variations in gut microbiome research on wildlife and domesticated animals in Southeast Asia. A comprehensive search of 91 relevant studies on the SCOPUS database yielded 54 suitable publications for review, encompassing diverse taxa such as invertebrates (20), fishes (7), reptiles (3), birds (5), and mammals (19). Notably, various methodological approaches were employed to characterize microbial communities, including the source of isolation, various culture-based approaches, sequencing methods, and the targeted markers. Based on the information provided in this study, future studies should strive to develop guidelines and best practices specific to gut microbiome studies. This would enhance comparability and facilitate the integration of findings. Such efforts will also advance our understanding of the microbial diversity associated with wildlife, and its potential implications for their health and conservation

The Asian region is globally renowned for its unparalleled biodiversity, rich ecosystems, and unique cultural heritage. However, the delicate equilibrium between human societies and the diverse flora and fauna faces increasing challenges due to the region's rich biodiversity and escalating utilization of wildlife resources. Hence, this paper aims to document the utilization of endangered wildlife across Asian countries and to report the existing conservation measures and recommendations for sustainable wildlife management over the years. We employed prominent academic databases, specifically SCOPUS, to explore the dynamics of wildlife utilization and its associated implications. From the findings, 9,989 records were successfully identified. However, only 65 articles were included in the review after the screening process. A total of 61 species, comprising 30 mammals, 22 reptiles, one amphibian, and eight birds, all endangered, were identified as subjects of wildlife utilization in 16 Asian countries. China possessed the highest count for wildlife utilization across all categories, including 15 species used for trading purposes. In conclusion, this research underscores the critical need for integrated approaches that balance human needs and conservation imperatives to ensure a sustainable future for Asia's wildlife biodiversity

Amphibians, a diverse and ecologically important group, are facing global declines due to various factors, including habitat loss and climate change. Accurate species identification is crucial for effective conservation efforts, and DNA barcoding has emerged as a powerful tool in this regard. This study compares the efficacy of two DNA barcoding primer sets, targeting the 16S ribosomal RNA gene and the Cytochrome Oxidase I (COI) gene, for identifying 20 amphibian species. While both primer sets successfully amplified sequences, the 16S rRNA gene region identified all 20 samples, whereas the COI region identified 14. The amplified sequences, approximately 550 base pairs for 16S rRNA and 658 base pairs for COI facilitated precise taxonomic placement within amphibian families using Neighbor-Joining phylogenetic trees. These findings enhance DNA barcoding methodology and aid in understanding amphibian diversity, crucial for effective conservation strategies amidst global declines drives by habitat loss, diseases, and climate change

The Asian region is globally renowned for its unparalleled biodiversity, rich ecosystems, and unique cultural heritage. However, the delicate equilibrium between human societies and the diverse flora and fauna faces increasing challenges due to the region's rich biodiversity and escalating utilization of wildlife resources. Hence, this paper aims to document the utilization of endangered wildlife across Asian countries and to report the existing conservation measures and recommendations for sustainable wildlife management over the years. We employed prominent academic databases, specifically SCOPUS, to explore the dynamics of wildlife utilization and its associated implications. From the findings, 9,989 records were successfully identified. However, only 65 articles were included in the review after the screening process. A total of 61 species, comprising 30 mammals, 22 reptiles, one amphibian, and eight birds, all endangered, were identified as subjects of wildlife utilization in 16 Asian countries. China possessed the highest count for wildlife utilization across all categories, including 15 species used for trading purposes. In conclusion, this research underscores the critical need for integrated approaches that balance human needs and conservation imperatives to ensure a sustainable future for Asia's wildlife biodiversity

Dietary assessment plays a crucial role in comprehending the ecological dynamics and nutritional needs of herbivores and omnivores. The metabarcoding technique has emerged as a potent tool for exploring the dietary composition of these animals. However, various metabarcoding techniques have been developed, each with its own advantages and limitations. This study aims to compare the performance of different metabarcoding techniques in herbivores and omnivores diet. We systematically reviewed 159 published manuscripts in Scopus and Google Scholar, and thematic analysis was conducted across several categories, including the marker, platform, and database utilized. Preliminary findings reveal significant variations among metabarcoding techniques across these two animal groups. The trnL gene exhibited higher taxonomic resolution for herbivorous species, whereas the combination of the 'trnL + 16s rRNA' gene exhibited superior performance for omnivorous species. The Illumina platform emerged as the most commonly used method for analyzing the diets of both herbivores and omnivores, with the primary reference database being the National Centre for Biotechnology Information (NCBI). This study offers valuable insights into the strengths and limitations of different metabarcoding techniques for dietary assessment in herbivores and omnivores and optimizing metabarcoding protocols, facilitating more precise and reliable diet analyses within these ecological groups

Next-generation sequencing (NGS) has transformed genomics, presenting significant potential for advancing wildlife conservation. NGS technology offers opportunities and challenges for addressing scientific questions in wildlife management. Unfortunately, there is a lack of comprehensive records on NGS implementation in Southeast Asia, particularly concerning wildlife conservation. To address this gap, we analyzed NGS studies focused on wildlife monitoring in Southeast Asia and introduced a practical framework for implementing NGS technologies in global wildlife conservation, especially in Southeast Asia. We systematically reviewed NGS studies in wildlife monitoring from the SCOPUS database. We identified 137 relevant publications from 11 countries, with Malaysia contributing 36% of the studies. The included studies were categorized into five themes: species identification, dietary assessment, health monitoring, taxonomic resolution, and whole-genome sequencing, with the majority focusing on 38 publications related to wildlife health. The framework developed in this study help the researchers and conservation practitioners with insights on NGS technology application in conservation while also addressing the benefits, limitations, and ethical considerations associated with NGS use. This review offers a brief overview of NGS usage and provides guidelines for embracing NGS as a valuable tool for effective wildlife conservation strategies in our rapidly changing world

The use of environmental DNA (eDNA) as the genetic tool to monitor biodiversity has been increasing across the world, including Malaysia. Over a decade, the technique has become widely used in wildlife conservation with the technologies called next-generation sequencing (NGS). Unfortunately, as one of the top megadiverse countries, Malaysia is still behind in this field since eDNA methods outperform traditional surveys. Hence, in this study, we analyzed the paper related to eDNA studies in Malaysia, which focused on wildlife monitoring. We conducted a systematic bibliographic search and quantitative review of studies published before June 2021 from Google Scholar and Scopus database. Forty published eDNA studies were found, and each paper was classified based on five themes (species identification, diet assessment, health monitoring, resolve taxonomic, whole-genome sequencing) according to the study analysis. This study aims to identify gaps in eDNA in Malaysia, which can later be useful for future conservation actions and development by applying technology in wildlife monitoring.

Recently, citizen scientists have gained recognition for engaging the public in collaboration with professional scientists in scientific actions. Citizen scientists act as an important force contributing to the Community, Education, and Public Awareness (CEPA) initiatives and promoting sustainable development. Thus, the study's objectives are to conduct a systematic review for citizen scientists in addressing the challenges of working in environmental conservation research and to address the way forward for fellow citizen scientists. We conducted a systematic literature review using the SCOPUS database in September 2023 to review the relatable article on citizen science. The result successfully extracted 118 related articles by applying a few keywords, ‘Conservation’, ‘Education’, ‘Environment’, ‘Management’, and ‘Citizen Scientist’ as a primary keyword. Notably, approximately 62% of articles published focused on environmental conservation involving animals and plants. This study also highlights the contributions of citizen scientists in conserving and monitoring biodiversity, including volunteering activities. In conclusion, this review emphasizes citizen scientists' transformative potential in supporting CEPA and offers insights into the crucial role of citizen scientists and their promising pathway toward sustainability

The use of environmental DNA (eDNA) as the genetic tool to monitor biodiversity has been increasing across the world, including Malaysia. Over a decade, the technique has become widely used in wildlife conservation with the technologies called next-generation sequencing (NGS). Unfortunately, as one of the top megadiverse countries, Malaysia is still behind in this field since eDNA methods outperform traditional surveys. Hence, in this study, we analyzed the paper related to eDNA studies in Malaysia, which focused on wildlife monitoring. We conducted a systematic bibliographic search and quantitative review of studies published before June 2021 from Google Scholar and Scopus database. Forty published eDNA studies were found, and each paper was classified based on five themes (species identification, diet assessment, health monitoring, resolve taxonomic, whole-genome sequencing) according to the study analysis. This study aims to identify gaps in eDNA in Malaysia, which can later be useful for future conservation actions and development by applying technology in wildlife monitoring.

Protected regions, such as Malaysia's Panti Forest Reserve (PFR), play a vital role in preserving freshwater biodiversity. However, the absence of baseline data on stream macroinvertebrates and sediment nutrient contents presents a significant challenge. This study, conducted from August 21-24, 2022, as part of the Panti Expedition organized by the Johor State Forestry Department, aimed to investigate the current nutrient flows and stream macroinvertebrates in PFR, providing preliminary assessments. Utilizing kick and grab sampling techniques, we collected stream macroinvertebrates and benthic sediments along the entire upstream-to-downstream gradient of Pelepah Kiri and Gunung Muntahak. Our findings revealed a diverse assemblage of 385 macroinvertebrates from 28 distinct families, with notable abundance observed in Baetidae (14.0%), Simuliidae (10.6%), and Hydropsychidae (10.6%). The analysis of overall taxonomic composition indicated a Jaccard index similarity of 0.64, suggesting overlapping communities between the two sites. Predators were significantly more abundant in Pelepah Kiri, while collectors were more abundant in Gunung Muntahak. The expedition trails exhibited varying levels of moisture content (ranging from 17.2% to 31.3%), total organic carbon content (≤ 0.4%), and total nitrogen levels (ranging from 1,330 mg/kg to 3,180 mg/kg), while phosphorus loading remained undetectable due to low concentration levels. Nutrient analyses indicated no significant impact of total nitrogen on macroinvertebrate family richness, abundance, and diversity. The study's biological metrics demonstrated good stream quality, with taxa richness ranging from 21 to 20 and EPT taxa richness ranging from 6 to 10. These results highlight the diverse freshwater macroinvertebrate communities in PFR, suggesting the maintenance of favorable water quality for biodiversity preservation. Overall, this study provides critical baseline data for future conservation management strategies in P

PrimaTourism is a well-known aspect of nature tourism that provides valuable benefits for the primate species conservation. Malaysia, with its rich diversity of more than 25 primate species inhabited in the country, holds significant potential for enhancing primate-based tourism. However, the information on primate species in potential tourism spots like Tasik Chini remains limited, with only a few data on their behaviour and ecology. Additionally, the lack of comprehensive information on the primate species poses challenges in identifying potential PrimaTourism product. Thus, this study aimed to assess primate diversity and explore the perception of PrimaTourism among local people in Tasik Chini. A survey and interview session were conducted over five days in two villages around Tasik Chini (Kampung Gumum and Felda Chini), involving a total of 32 respondents. Most respondents reported the presence of two common primate species in their area: the long-tailed macaque (Macaca fascicularis) and the pig-tailed macaque (Macaca nemestrina). The result suggested a significant majority (81%) of respondents expressed support for the dusky leaf monkey (Trachypithecus obscurus) as an ideal primate species for PrimaTourism in Tasik Chini, citing its attractive coloration and favoring the development of primate-based tourism. Overall, these findings can serve as essential baseline data for implementing primate tourism in the area, engaging the local community to attract tourists to Tasik Chini effectively and eventually serve as a conservation tool for the primate species.

Due to the high demand in the fisheries market, species identification is vital for commercial fish to avoid mislabeling and fraud in marketplaces. The increasingly threatening human activities like overfishing, blast fishing, poison fishing, and trawling have become the major threats to fish and caused an urgent need for documentation of both marine and freshwater fishes. Commercial fish products sometimes are sold by their parts, such as fins which further create confusion for identification purposes. Morphological identification also requires highly skilled ichthyologists to avoid misidentification. Thus, identification through DNA barcoding can be utilized as it holds the potential for accurate and rapid identification. Here we identified the commercial marine fish species sold in Muar Fish Market, Johor, using the Cytochrome Oxidase subunit I (COI) gene of mitochondrial DNA (mtDNA). A 700 base-pair sequence of the COI region was targeted by amplifying extracted DNA from 28 fin samples collected from the fish market. From 28 samples, we successfully sequenced 16 samples, and by using phylogenetic analysis, (Neighbor-Joining (NJ), Maximum Parsimony (MP) and Bayesian Inference (BI)) we successfully identified the samples belonging to nine families (Scombridae, Platycephalidae, Carangidae, Ariidae, Polynemidae, Coryphaenidae, Sphyraenidae, Muraenesocidae, Engraiulidae), 12 genera and 16 species. This study reports the first documentation of commercial fish sold in Muar through the DNA barcoding technique. This technique should be expanded further to identify any species with important conservation implications, such as endangered species, for improved management of fisheries in Malaysia.