High-throughput DNA metabarcoding for determining the gut microbiome of captive critically endangered Malayan tiger (Panthera tigris jacksoni) during fasting

The Malayan tiger (Panthera tigris jacksoni) is a critically endangered species native to the Malaysian Peninsula. To imitate wild conditions where tigers do not hunt every day, numerous wildlife sanctuaries do not feed their tigers daily. However, the effects of fasting on the gut microbiota of captive Malayan tigers remains unknown. This study aimed to characterise the gut microbiota of captive Malayan tigers by comparing their microbial communities during fasting versus normal feeding conditions. This study was conducted at the Melaka Zoo, Malaysian Peninsula and involved Malayan tigers fasted every Monday. In total, ten faecal samples of Malayan tiger, two of Bengal tiger (outgroup) and four of lion (outgroup) were collected and analysed for metabarcoding targeting the 16S rRNA V3–V4 region. In total, we determined 14 phyla, 87 families, 167 genera and 53 species of gut microbiome across Malayan tiger samples. The potentially harmful bacterial genera found in this study included Fusobacterium, Bacteroides, Clostridium sensu stricto 1,Solobacterium, Echerichia shigella, Ignatzschineria and Negativibacillus. The microbiome in the fasting phase had a higher composition and was more diverse than in the feeding phase. The present findings indicate a balanced ratio in the dominant phyla, reflecting a resetting of the imbalanced gut microbiota due to fasting. These findings can help authorities in how to best maintain and improve the husbandry and health of Malayan tigers in captivity and be used for monitoring in ex-situ veterinary care unit

Determining the diet of wild Asian elephants (Elephas maximus) at human–elephant conflict areas in Peninsular Malaysia using DNA metabarcoding

Human–elephant conflict (HEC) contributes to the increasing death of Asian elephants due to road accidents, retaliatory killings and fatal infections from being trapped in snares. Understanding the diet of elephants throughout Peninsular Malaysia remains crucial to improve their habitat quality and reduce scenarios of HEC. DNA metabarcoding allows investigating the diet of animals without direct observation, especially in risky conflict areas. The aim of this study was to determine: i) the diet of wild Asian elephants from HEC areas in Peninsular Malaysia using DNA metabarcoding and ii) the influence of distinct environmental parameters at HEC locations on their feeding patterns. DNA was extracted from 39 faecal samples and pooled into 12 groups representing the different sample locations: Kuala Koh, Kenyir, Ulu Muda, Sira Batu, Kupang-Grik, Bumbun Tahan, Belum-Temengor, Grik, Kampung Pagi, Kampung Kuala Balah, Aring 10 and the National Elephant Conservation Centre, which served as a positive control for this study. DNA amplification and sequencing targeted the ribulose-bisphosphate carboxylase gene using the next-generation sequencing Illumina iSeq100 platform. Overall, we identified 35 orders, 88 families, 196 genera and 237 species of plants in the diet of the Asian elephants at HEC hotspots. Ficus (Moraceae), Curcuma (Zingiberaceae), Phoenix (Arecaceae), Maackia (Fabaceae), Garcinia (Clusiaceae) and Dichapetalum (Dichapetalaceae) were the highly abundant dietary plants. The plants successfully identified in this study could be used by the Department of Wildlife and National Parks (PERHILITAN) to create buffer zones by planting the recommended dietary plants around HEC locations and trails of elephants within Central Forest Spine (CFS) landscape

Haplotype analysis of long-tailed macaques in TNB Bukit Selambau, Peninsular Malaysia, leads to inferences of fissionfusion social structure

Long-tailed macaques have been roaming near Tenaga National Berhad Bukit Selambau Solar(TBSS) causing human–macaque conflicts. This study reveals the social organization and genetic variation of these macaques. Macaques’ groups were determined via direct observation and closed circuit television. Genomic DNA from 29 fecal samples were extracted and proceeded with amplification of the D-loop region of mitochondrial DNA. There are four main groups of primates at TBSS. Some members of the Kuil group and the Pekan group shared haplotype 1, forming a fission-fusion society. This finding can be used for TNB in mitigation plans involving human– macaque conflict

De novo sequencing, assembly and analysis of eight different transcriptomes from the Malayan pangolin

Pangolins are scale-covered mammals, containing eight endangered species. Maintaining pangolins in captivity is a significant challenge, in part because little is known about their genetics. Here we provide the first large-scale sequencing of the critically endangered Manis javanica transcriptomes from eight different organs using Illumina HiSeq technology, yielding ~75 Giga bases and 89,754 unigenes. We found some unigenes involved in the insect hormone biosynthesis pathway and also 747 lipids metabolism-related unigenes that may be insightful to understand the lipid metabolism system in pangolins. Comparative analysis between M. javanica and other mammals revealed many pangolin-specific genes significantly over-represented in stress-related processes, cell proliferation and external stimulus, probably reflecting the traits and adaptations of the analyzed pregnant female M. javanica. Our study provides an invaluable resource for future functional works that may be highly relevant for the conservation of pangolins

Pangolin genomes and the evolution of mammalian scales and immunity

Pangolins, unique mammals with scales over most of their body, no teeth, poor vision, and an acute olfactory system, comprise the only placental order (Pholidota) without a whole-genome map. To investigate pangolin biology and evolution, we developed genome assemblies of the Malayan (Manis javanica) and Chinese (M. pentadactyla) pangolins. Strikingly, we found that interferon epsilon (IFNE), exclusively expressed in epithelial cells and important in skin and mucosal immunity, is pseudogenized in all African and Asian pangolin species that we examined, perhaps impacting resistance to infection. We propose that scale development was an innovation that provided protection against injuries or stress and reduced pangolin vulnerability to infection. Further evidence of specialized adaptations was evident from positively selected genes involving immunity-related pathways, inflammation, energy storage and metabolism, muscular and nervous systems, and scale/hair development. Olfactory receptor gene families are significantly expanded in pangolins, reflecting their well-developed olfaction system. This study provides insights into mammalian adaptation and functional diversification, new research tools and questions, and perhaps a new natural IFNE-deficient animal model for studying mammalian immunity.University of Malaya and Ministry of Education, Malaysia [UM.C/HIR/MOHE/08]; UMRG grant from the University of Malaya and Ministry of Education, Malaysia [RG541-13HTM]; Russian Ministry of Science [11.G34.31.0068]; NIH-NHGRI grant [5U54HG00307907]SCI(E)[email protected]

Population genetics and evaluation of diallel cross of four red tilapia populations in Malaysia / Kayal Vizi a/p Karuppannan

Tilapia is an important farmed freshwater fish in Malaysia. Tilapia farming was introduced in Malaysia in the 1940’s. In 2003, tilapia accounted 21,417 tons of total freshwater aquaculture production. Red hybrid is the most popular tilapia species cultured in Malaysia. One of the constraints faced by the industry is quality of fries. There are quite a number of private hatcheries in Malaysia producing tilapia fries and not much is known about their quality with respect to their genetic variability. Thus this study attempted to characterize the four study populations and one standard population, which were PKPS (PS), Bentong (BT), Enggor (EG), Negeri Sembilan (NS) and Gift (GT) as control population using 20 polymorhic Microsatellite markers. Gift population is Nile tilapia. Their dendogram relationship showed that the hetrozygosity value was high, which indicated the presence of high genetic variability among the five populations. As a continuation, Diallel Crossing was started among four populations, which were PKPS (PS), Bentong (BN), Enggor (EG) and Negeri Sembilan (NS). Attention given to two traits, which were Growth and Survival. In Growth Factor, trait studied was Specific Growth Rate. In Survival, Egg Hatchability, Fry Survival and Adult Fish Survival were the three parameters focused on. As a outcome of this study, managed to produce broodstocks with known genetic attributes through quantitative genetics for fast growth rates based on high heritability, high additive variance and low environmental effects

High-throughput DNA metabarcoding for determining the gut microbiome of captive critically endangered Malayan tiger (Panthera tigris jacksoni) during fasting

The Malayan tiger (Panthera tigris jacksoni) is a critically endangered species native to the Malaysian Peninsula. To imitate wild conditions where tigers do not hunt every day, numerous wildlife sanctuaries do not feed their tigers daily. However, the effects of fasting on the gut microbiota of captive Malayan tigers remains unknown. This study aimed to characterise the gut microbiota of captive Malayan tigers by comparing their microbial communities during fasting versus normal feeding conditions. This study was conducted at the Melaka Zoo, Malaysian Peninsula and involved Malayan tigers fasted every Monday. In total, ten faecal samples of Malayan tiger, two of Bengal tiger (outgroup) and four of lion (outgroup) were collected and analysed for metabarcoding targeting the 16S rRNA V3–V4 region. In total, we determined 14 phyla, 87 families, 167 genera and 53 species of gut microbiome across Malayan tiger samples. The potentially harmful bacterial genera found in this study included Fusobacterium, Bacteroides, Clostridium sensu stricto 1, Solobacterium, Echerichia shigella, Ignatzschineria and Negativibacillus. The microbiome in the fasting phase had a higher composition and was more diverse than in the feeding phase. The present findings indicate a balanced ratio in the dominant phyla, reflecting a resetting of the imbalanced gut microbiota due to fasting. These findings can help authorities in how to best maintain and improve the husbandry and health of Malayan tigers in captivity and be used for monitoring in ex-situ veterinary care unit

Metabarcoding data analysis revealed the plant dietary variation of long-tailed macaque Macaca fascicularis (Cercopithecidae, Cercopithecinae) living in disturbed habitats in Peninsular Malaysia

The long-tailed macaque (Macaca fascicularis) has a wide range in both Peninsular Malaysia and Borneo. Although the primates are especially vulnerable to habitat alterations, this primate lives in disturbed habitats due to human-induced land-use. Thus, this study presents a faecal metabarcoding approach to clarify the plant diet of long-tailed macaques from five locations in Peninsular Malaysia to represent fragmented forest, forest edge, island and recreational park habitats. We extracted genomic DNA from 53 long-tailmacaque faecal samples. We found 47 orders, 126 families, 609 genera and 818 species across these five localities. A total of 113 plant families were consumed by long-tailed macaques in Universiti Kebangsaan Malaysia, 61 in the Malaysia Genome and Vaccine Institute, 33 in Langkawi Island, 53 in Redang Island and 44 in the Cenderawasih Cave. Moraceae (33.24%) and Fabaceae (13.63%) were the most common families consumed by long-tailed macaques from the study localities. We found that habitat type impacted diet composition, indicating the flexibility of foraging activities. This research findings provide an understanding of plant dietary diversity and the adaptability of this macaque with the current alteration level that applies to long-tailed macaque conservation management interest in the future. Keywords Malaysian primates, Southeast Asia, DNA metabarcoding, trnL, next-generationsequencin

Metabarcoding data analysis revealed the plant dietary variation of long-tailed macaque Macaca fascicularis (Cercopithecidae, Cercopithecinae) living in disturbed habitats in Peninsular Malaysia

The long-tailed macaque (Macaca fascicularis) has a wide range in both Peninsular Malaysia and Borneo. Although the primates are especially vulnerable to habitat alterations, this primate lives in disturbed habitats due to human-induced land-use. Thus, this study presents a faecal metabarcoding approach to clarify the plant diet of long-tailed macaques from five locations in Peninsular Malaysia to represent fragmented forest, forest edge, island and recreational park habitats. We extracted genomic DNA from 53 long-tailed macaque faecal samples. We found 47 orders, 126 families, 609 genera and 818 species across these five localities. A total of 113 plant families were consumed by long-tailed macaques in Universiti Kebangsaan Malaysia, 61 in the Malaysia Genome and Vaccine Institute, 33 in Langkawi Island, 53 in Redang Island and 44 in the Cenderawasih Cave. Moraceae (33.24%) and Fabaceae (13.63%) were the most common families consumed by long-tailed macaques from the study localities. We found that habitat type impacted diet composition, indicating the flexibility of foraging activities. This research findings provide an understanding of plant dietary diversity and the adaptability of this macaque with the current alteration level that applies to long-tailed macaque conservation management interest in the future

Haplotype analysis of long-tailed macaques in TNB Bukit Selambau, Peninsular Malaysia, leads to inferences of fissionfusion social structure

Long-tailed macaques have been roaming near Tenaga National Berhad Bukit Selambau Solar (TBSS) causing human–macaque conflicts. This study reveals the social organization and genetic variation of these macaques. Macaques’ groups were determined via direct observation and closedcircuit television. Genomic DNA from 29 fecal samples were extracted and proceeded with amplification of the D-loop region of mitochondrial DNA. There are four main groups of primates at TBSS. Some members of the Kuil group and the Pekan group shared haplotype 1, forming a fission-fusion society. This finding can be used for TNB in mitigation plans involving human– macaque conflict