Successful captive breeding of a Malayan pangolin population to the third filial generation

Pangolins are threatened placental mammals distributed in Africa and Asia. Many efforts have been undertaken in the last century to maintain pangolins in captivity, but only a few of them succeeded in maintaining and keeping this species in a controlled environment. This study reports the first systematic breeding of the Critically Endangered Malayan pangolin (Manis javanica) in captivity. Our captive breeding approach successfully improved the reproductive rate for both wild and captive-born female pangolins. From 2016 to 2020, we had 33 wild pangolins and produced 49 captive-born offspring spanning three filial generations. The female offspring further bred 18 offspring, of which 14 (78%) were conceived during the first time of cohabitation with males, and four offspring were conceived during the second cohabitation event, suggesting that they may practice copulation-induced ovulation. We observed that captive-born female pangolins could reach sexual maturity at 7–9 months (n = 4), and male pangolins could mate and successfully fertilise females at nine months age (n = 1). We also observed a female pangolin conceiving on the eighth day after parturition (the fifth day after the death of its pup). Our captive pangolins had a female-biased sex ratio of 1:0.5 at birth, unlike other known captive-born mammals. Also, captive-born pangolins were generally more viable after successful weaning and had a similar gestation length (~185 days) to wild pangolins. Most importantly, we report the first self-sustaining captive population of Malayan pangolins, and this species has an efficient reproduction strategy. These advances provide more comprehensive information for people to understand pangolins, and have implications for conserving endangered Malayan pangolins and providing scientific guidance to the management of other pangolin species.

First report on detection of Babesia spp. in confiscated Sunda pangolins (Manis javanica) in Thailand

Background and Aim: The Sunda pangolin (Manis javanica) is on the International Union for Conservation of Nature Red List of Threatened Species (critically endangered) due to high levels of illegal trafficking for its products. Thailand is one of the habitats of this species, and it has become the main hub for its illegal trafficking. Rehabilitating these captive pangolins and reintroducing them back to the wild are challenging due to the limited knowledge on their diet, management, and diseases. Hemoparasites, including Babesia spp., can cause important protozoal infections in both domestic and wild animals, resulting in the failure of rehabilitation and conservation programs. However, Babesia spp. has not been reported in pangolins. The aim of the study was to determine the prevalence of Babesia spp. in the Sunda pangolin of Thailand. Materials and Methods: A total of 128 confiscated Sunda pangolins from across different regions in Thailand were investigated. These pangolins had been admitted to a regional Wildlife Quarantine Center for rehabilitation before release in the forest. Routine physical examinations were conducted on the animals. We collected blood samples from each pangolin for hematological analysis and to detect Babesia spp. using polymerase chain reaction (PCR) targeting the partial 18s rRNA gene. Results: Babesia-specific PCR detected 53 animals (41.4%) that were positive for Babesia spp. Blood smears were obtained from the positive samples and investigated under a light microscope to observe for trophozoites of Babesia spp. Examination of 40 PCR-positive and -negative samples found no significant differences between the hematological parameters of Babesia-positive and Babesia-negative samples. Eight PCR-positive samples were randomly selected and their DNA was sequenced. Seven and one of sequences match uncharacterized Babesia spp. with 100% and 99.2% similarity, respectively. Phylogenetic analysis demonstrated that our samples form a unique monophyletic clade along with other Babesia spp. detected in the wild. This clade is clearly separated from other Babesia spp. from small carnivores, ruminants, and rats. Conclusion: Our results provide evidence of infection of Sunda pangolins in Thailand by Babesia spp. These pangolins originated from different regions and had not lived together before blood collection. Thus, we suggest that the uncharacterized Babesia spp. found in this study constitute a new group of pangolin-specific Babesia spp. The prevalence of the uncharacterized Babesia spp. was not correlated to pangolin health. Further studies are required to characterize the genomes and phenotypes, including the morphology and pathogenicity of these protozoa. Such information will be helpful for the conservation and health management of the Sunda pangolin.

ANALISIS KINERJA PENEGAKAN HUKUM DALAM MENURUNKAN KASUS PERDAGANGAN ILLEGAL SATWA YANG DILINDUNGI DI PROVINSI KALIMANTAN BARAT

This research is motivated by the high cases of illegal trade of protected wildlife in West Kalimantan Province. Animal trade tends to be endangered and included in the endangered list. Actions of animal trade that still occur in West Kalimantan Province indicate the low level of law enforcement and weak supervision by related parties in combating cases of protected animal trade. The purpose of this study is to examine the factors causing the occurrence of illegal trade crimes against protected animals in the West Kalimantan region, and analyze the performance of stakeholders in law enforcement against the illegal trade of protected animals in the West Kalimantan region based on their role and authority. This study uses a survey method that is guided by a questionnaire. The selection of respondents by purposive sampling, data analysis using policy analysis then the data is descriptive with tabulation. The results of the study show that in the period of 2016-2019 there were 49 cases of illegal wildlife trade in West Kalimantan Province, the traded animals are species belonging to protected animals and classified in the Appendix I. category. The dominant protected species traded in the 2016-2019 period originated from anteater species (Manis javanica), Orang-utan (Pongo pygmaeus), and Turtle (Chelonia sp.). The dominant factor in the illegal trade of protected animals in the province of West Kalimantan in the 2016-2019 period was economic factors, followed by motives for the collection of rare goods and consumption. The performance of stakeholders in law enforcement against protected animals in the West Kalimantan region has succeeded in handling criminal cases against protected animals, all of which ended with the execution of the defendant.Keywords: law enforcement performance, protected animals, illegal trading, stakeholders.AbstrakPenelitian ini dilatarbelakangi tingginya kasus perdagangan illegal satwa dilindungi di Provinsi Kalimantan Barat. Perdagangan satwa yang dilakukan cenderung pada spesies-spesies langka dan termasuk dalam daftar terancam punah. Aksi perdagangan satwa yang masih terjadi di Provinsi Kalimantan Barat mengindikasikan masih rendahnya penegakan hukum dan lemahnya pengawasan oleh pihak terkait dalam memberantas kasus perdagangan satwa yang dilindungi. Tujuan penelitian ini untuk mengkaji faktor penyebab terjadinya kejahatan perdagangan illegal terhadap satwa yang dilindungi, serta menganalisis kinerja pemangku kepentingan dalam penegakan hukum terhadap perdagangan illegal satwa yang dilindungi di wilayah Kalimantan Barat berdasarkan peran dan kewenangan. Penelitian ini menggunakan metode survey yang dipandu dengan kuesioner. Pemilihan responden dengan purposive sampling, analisis data menggunakan analisis kebijakan kemudian data dideskriptif dengan tabulasi. Hasil penelitian menunjukan kurun waktu 2016-2019 terdapat 49 kasus perdagangan satwa illegal di Provinsi Kalimantan Barat, satwa yang diperdagangkan merupakan jenis satwa yang termasuk ke dalam satwa dilindungi dan tergolong dalam kateogri Appindiks I. Satwa dilindungi dominan yang diperdagangkan pada periode 2016-2019 berasal dari spesies trenggiling (Manis javanica), Orang utan (Pongo pygmeus), dan Penyu (Chelonia sp.). Faktor dominan perdagangan illegal satwa dilindungi di Provinsi Kalimantan Barat dalam periode 2016-2019 adalah faktor ekonomi kemudian diikuti motif koleksi barang langka dan konsumsi. Kinerja pemangku kepentingan dalam penegakan hukum terhadap satwa yang dilindungi di wilayah Kalimantan Barat, telah berhasil menangani kasus pidana terhadap satwa dilindungi yang kesemuanya berahir dengan eksekusi kepada terdakwa.Kata kunci: kinerja penegakan hukum, perdagangan illegal, satwa dilindungi, pemangku kepentingan.

Molecular Detection of Novel Borrelia Species, Candidatus Borrelia javanense, in Amblyomma javanense Ticks from Pangolins

A novel Borrelia species, Candidatus Borrelia javanense, was found in ectoparasite ticks, Amblyomma javanense, from Manis javanica pangolins seized in anti-smuggling operations in southern China. Overall, 12 tick samples in 227 (overall prevalence 5.3%) were positive for Candidatus B. javanense, 9 (5.1%) in 176 males, and 3 (5.9%) in 51 females. The phylogenetic analysis, based on the 16S rRNA gene and the flagellin gene sequences of the Borrelia sp., exhibited strong evidence that Candidatus B. javanense did not belong to the Lyme disease Borrelia group and the relapsing fever Borrelia group but another lineage of Borrelia. The discovery of the novel Borrelia species suggests that A. javanense may be the transmit vector, and the M. javanica pangolins should be considered a possible origin reservoir in the natural circulation of these new pathogens. To our knowledge, this is the first identification of a novel Borrelia species agent in A. javanense from pangolins. Whether the novel agent is pathogenic to humans is unknown and needs further research.

The gut microbiome of the Sunda pangolin (Manis javanica) reveals its adaptation to specialized myrmecophagy

Background The gut microbiomes of mammals are closely related to the diets of their hosts. The Sunda pangolin (Manis javanica) is a specialized myrmecophage, but its gut microbiome has rarely been studied. Methods Using high-throughput Illumina barcoded 16S rRNA amplicons of nine fecal samples from nine captive Sunda pangolins, we investigated their gut microbiomes. Results The detected bacteria were classified into 14 phyla, 24 classes, 48 orders, 97 families, and 271 genera. The main bacterial phyla were Firmicutes (73.71%), Proteobacteria (18.42%), Actinobacteria (3.44%), and Bacteroidetes (0.51%). In the PCoA and neighbor-net network (PERMANOVA: pangolins vs. other diets, weighted UniFrac distance p < 0.01, unweighted UniFrac distance p < 0.001), the gut microbiomes of the Sunda pangolins were distinct from those of mammals with different diets, but were much closer to other myrmecophages, and to carnivores, while distant from herbivores. We identified some gut microbiomes related to the digestion of chitin, including Lactococcus, Bacteroides, Bacillus, and Staphylococcus species, which confirms that the gut microbiome of pangolins may help them to digest chitin. Significance The results will aid studies of extreme dietary adaption and the mechanisms of diet differentiation in mammals, as well as metagenomic studies, captive breeding, and ex situ conservation of pangolins.

Predicting the Distribution of Sunda Pangolin (Manis javanica Desmarest, 1822) in Way Canguk Research Station, Bukit Barisan Selatan National Park, Lampung

The distribution of a species can help guide the protection activities in their natural habitat. Conversely, the lack of information on this distribution makes the protection strategy of this species difficult. The research was conducted in Way Canguk Research Station, Bukit Barisan Selatan National Park from January until March 2018. The purposes of this research were to create a distribution prediction map of Sunda pangolin (Manis javanica) and estimating the environment variables that most influenced the probability of the distribution. Fourteen points of camera trap coordinates were used for presence data with nine types of environment variables such as elevation, slope, understorey, canopy cover, distance from roads, distance from rivers, distance from villages, food source, and distance from the threat. The result of maxent showed an Area Under the Curve (AUC) value of 0.909 categorized as very good. The highest probability of Sunda pangolin distributions was in the Pemerihan Resort and Way Haru Resort area, while the dominant environmental variables included the distance from the village, the canopy cover, and the distance from threat with the value 47.7; 25.85; and 15.8%, respectively. Prediction maps and environment variables can help to identify the population of Sunda pangolin in the wild and can provide input for the national parks to prioritize protection areas for Sunda pangolin from the increased poaching.

Pathogenicity and transmissibility of a novel respirovirus isolated from a Malayan pangolin

The identification of SARS-CoV-2-like viruses in Malayan pangolins (Manis javanica) has focused attention on these endangered animals and the viruses they carry. We successfully isolated a novel respirovirus from the lungs of a dead Malayan pangolin. Similar to murine respirovirus, the full-length genome of this novel virus was 15 384 nucleotides comprising six genes in the order 3′–(leader)–NP-P-M-F-HN-l-(trailer)−5’. Phylogenetic analysis revealed that this virus belongs to the genus Respirovirus and is most closely related to murine respirovirus. Notably, animal infection experiments indicated that the pangolin virus is highly pathogenic and transmissible in mice, with inoculated mice having variable clinical symptoms and a fatality rate of 70.37 %. The virus was found to replicate in most tissues with the exception of muscle and heart. Contact transmission of the virus was 100 % efficient, although the mice in the contact group displayed milder symptoms, with the virus mainly being detected in the trachea and lungs. The isolation of a novel respirovirus from the Malayan pangolin provides new insight into the evolution and distribution of this important group of viruses and again demonstrates the potential infectious disease threats faced by endangered pangolins.