Identification of CITES-Listed Euphorbia royleana through DNA Barcoding Technology: A New Facet in Wildlife Forensics

Disorganized and chaotic collection of the Euphorbia plant species from the wild is one of the major reasons for its endangered status. According to CITES, the trade in Euphorbia royleana species is prohibited under Appendix II. However, the trade continues unabated as current identification methods do not discriminate between closely related species. In the present study, a DNA barcoding method has been used to establish inter- and intra-specific divergences of both matK and rbcL regions by using pairwise genetic distance measurement methods for evaluating the maximum barcoding gap. The matk and rbcL yielded a 100% amplification and sequencing success rate to distinguish closely related species of Euphorbia royleana unambiguously. The matk and rbcL showed average interspecific genetic distance divergence values of 0.031and 0.015, respectively. The maximum number of species-specific SNPs was observed in matK sequences at seven consecutive sites, which could distinguish Euphorbia royleana from closely related species. The best candidate barcoding region to identify Euphorbia royleana was found to be matK with a single-locus barcoding approach. Furthermore, the species discrimination method was developed with the help of species-specific SNPs derived from the matK barcoding region to accurately authenticate Euphorbia royleana, and it provided 100% species resolution

High-resolution geophysical imaging of reptile burrows (San Salvador rock iguana, The Bahamas): implications for ichnology and conservation ecology

Neoichnological research of terrestrial tracemakers in coastal settings provides important palaeoenvironmental information about their context within the subaerial facies. Here we present the first geophysical dataset of reptile burrows in a carbonate substrate and use it to help visualize parts of the burrows of the Bahamian (San Salvador) rock iguana (Cyclura rileyi). High-resolution 800 MHz ground-penetrating radar (GPR) images within an enclosure on San Salvador Island were employed to discriminate between the electromagnetic signal response from subsurface anomalies related to air-dominated voids or live animals within burrows. The dielectric contrast between the carbonate substrate and open burrows was sufficient to identify the majority of 15-20-cm-wide subsurface extensions of the inclined tunnels in the upper 30-40 cm. Whereas limestone clasts induced some interference, it is possible to differentiate their high-amplitude diffractions from those produced by the iguana burrows. Our research indicates that GPR imaging is a viable, rapid, non-invasive method of visualizing animal burrows, with implications to neoichnology, paleoichnology, and conservation ecology of semi-fossorial species. Furthermore, the critically endangered status of Bahamian land iguanas, as well as ongoing threats from natural and introduced pressures, highlights the need for research into their ichnological record.

Genetic Diversity and Population Structure Revealed by SSR Markers on Endemic Species Osmanthus serrulatus Rehder from Southwestern Sichuan Basin, China

Osmanthus serrulatus Rehder (Oleaceae) is an endemic spring-flowering species in China. It is narrowly distributed in the southwestern Sichuan Basin, and is facing the unprecedented threat of extinction due to problems associated with natural regeneration, habitat fragmentation and persistent and serious human interference. Here, the genetic diversity and population structure of 262 individuals from ten natural populations were analyzed using 18 microsatellites (SSR) markers. In total, 465 alleles were detected across 262 individuals, with a high polymorphic information content (PIC = 0.893). A high level of genetic diversity was inferred from the genetic diversity parameters (He = 0.694, I = 1.492 and PPL = 98.33%). AMOVA showed that a 21.55% genetic variation existed among populations and the mean pairwise Fst (0.215) indicated moderate genetic population differentiation. The ten populations were basically divided into three groups, including two obviously independent groups. Our results indicate that multiple factors were responsible for the complicated genetic relationship and endangered status of O. serrulatus. The concentrated distribution seems to be the key factor causing endangerment, and poor regeneration, human-induced habitat loss and fragmentation seem to be the primary factors in the population decline and further genetic diversity loss. These findings will assist in future conservation management and the scientific breeding of O. serrulatus.

Optimization of reproduction of ricefish endemic to Southeast Sulawesi Oryzias woworae Parenti & Hadiaty, 2010 through different sex ratios in spawning

Oryzias woworae has a beautiful color and has been traded as ornamental fish. This species is an endemic ricefish from Muna Island, Southeast Sulawesi, and is threatened with endangered status. Information on the optimal spawning sex ratio in O. woworae is unknown. This study aimed was to examine the optimization of reproduction based on the sex ratio of male: female broodstock O. woworae involved in spawning. The sex ratio of male: female spawning broodstock used were 1:1, 1:2, 1:3, and 1:4. The male and female brooders used had a total length of 3,1 ± 0,5 cm and 2,5 ± 0,5 cm. After adaptation, the broodstock of O. woworae was put into aquariums according to the treatment, and each container contained three spawning substrates. The harvesting of eggs on the substrate is carried out two times a day. The results showed that spawning O. woworae with a ratio of 1♂: 4♀ produced the lowest number of eggs and was significantly different with ratios of 1♂: 1♀ and 1♂: 2♀ (P<0,05). The ratio of 1♂ : 3♀ gave the highest egg hatching of 55% but was not significantly different from other treatments (P>0,05). The difference in male and female ratios did not affect the survival rate of O. woworae larvae (P>0,05), with values from 91,9-100%. The highest larvae produced was found in the spawning ratio of 1♂ : 3♀ with 37 larvae but not significantly different from other treatments (P>0,05). The water quality values during the study were temperature 26,5-310C, pH 5,5-8,8, and dissolved oxygen 5,3-6,0 mg L-1. O. woworae broodstock spawning can be optimized with a male to female ratio of 1:3.

Genetic Diversity of 17 Autochthonous Italian Chicken Breeds and Their Extinction Risk Status

The preservation of genetic variability of autochthonous poultry breeds is crucial in global biodiversity. A recent report revealed small breed size and potential risk of extinction of all native Italian poultry breeds; therefore, a correct assessment of their genetic diversity is necessary for a suitable management of their preservation. In this work, we provided an overview of the contribution to poultry biodiversity of some Italian autochthonous breeds reared in conservation centers devoted to local biodiversity preservation. The level of genetic diversity, molecular kinship, inbreeding, contribution to overall genetic diversity, and rate of extinction of each breed were analyzed with a set of 14 microsatellite loci in 17 autochthonous chicken breeds. To evaluate genetic variability, total number (Na), and effective number (Ne) of alleles, observed (Ho) and expected (He) heterozygosity, and F (Wright’s inbreeding coefficient) index were surveyed. The contribution of each analyzed breed to genetic diversity of the whole dataset was assessed using MolKin3.0; global genetic diversity and allelic richness contributions were evaluated. All the investigated loci were polymorphic; 209 alleles were identified (94 of which private alleles). The average number of alleles per locus was 3.62, and the effective number of alleles was 2.27. The Ne resulted lower in all breeds due to the presence of low-frequency alleles that can be easily lost by genetic drift, thus reducing the genetic variability of the breeds, and increasing their risk of extinction. The global molecular kinship was 27%, the average breed molecular kinship was 53%, and the mean inbreeding rate 43%, with a self-coancestry of 78%. Wright’s statistical analysis showed a 41% excess of homozygous due to breed genetic differences (34%) and to inbreeding within the breed (9%). Genetic variability analysis showed that 11 breeds were in endangered status. The contribution to Italian poultry genetic diversity, estimated as global genetic diversity, and ranged from 30.2 to 98.5%. In conclusion, the investigated breeds maintain a unique genetic pattern and play an important role in global Italian poultry biodiversity, providing a remarkable contribution to genetic variability.

The differences of body measurements in captive Sumatran elephant on different captivity management and sex

Wibowo DS, Ibrahim A, Widayanti R, Nurcahyo W, Wahyu M, Michaux J, Wijayanto H. 2021. The differences of body measurements in captive Sumatran elephant on different captivity management and sex. Biodiversitas 22: 2619-2624. Sumatran elephants are one of wild animals in Indonesia that currently received critically endangered status. This study aimed to measure the body size of both sexes of Sumatran elephants in the semi-captive (ECCs) and full captive (Zoo) management for conservation purposes. This study was conducted in ECCs in Sumatran island and Zoo on Java island. Body measurements (Chest Girth, Length, Height, Tarsal and Carpal Circumference) were collected from a total of 49 samples of captive Sumatran elephants (25 from EECs and 24 from Zoo). Data were analyzed using a completely randomized factorial design with different management and sex as the factors. The results showed that BL (304.92 vs 311.13) and TC (76.44 vs 77.50) were significantly different (P<0.05) across captivity managements. Sex categories (male and female) showed significant results (P<0.01) on CG (321.71 vs 302.46), BL (303.50 vs 272.49), BH (230.14 vs 208.23) and TC (82.29 vs 74.83). No interaction was identified between captive management and sex on body measurements. In conclusion, there were significant differences in body size of male and female elephants. This result contributed well-researched information to evaluate the conservation management based on body size and for further research on Sumatran elephant conservation.

Conserving an Endangered Canid: Assessing Distribution, Habitat Protection, and Connectivity for the Dhole (Cuon alpinus) in Bhutan

Most canids face population declines and range contractions worldwide. Although the dhole (Cuon alpinus) is widely distributed across 10 countries in South and Southeast Asia, limited studies exist on this species. Despite its globally “Endangered” status and ecological role as an apex predator, assessments on its distribution are limited to a few landscapes and countries. This explains the lack of a dhole-specific species conservation plan in most range countries, including Bhutan where no current population estimate exists. The species has also recovered from a country-wide poisoning campaign in the 1970s and 80s. In this study, we determine the dhole's distribution pattern and assess the protection and connectivity of dhole habitat in Bhutan. We anticipated dholes to be extant within their habitat well-represented in protected areas (PAs) and biological corridors (BCs). We used 721 georeferenced dhole occurrence records and eight environmental variables in MaxEnt software to model potential dhole distribution and habitat suitability. The model output was overlaid on the spatial layers of PAs and BCs to assess habitat protection and connectivity. As anticipated, we found the dhole widely distributed in all districts, PAs, and BCs in Bhutan. Dholes were recorded at the highest elevation range limit of 4,980 m above sea level, which overlapped with the “Vulnerable” snow leopard (Panthera uncia). Our model identified 72% (27,634 km2) of the country as suitable areas for dholes, of which, 31% (11,899 km2) was highly suitable and 41% (15,735 km2) was moderately suitable. Contrary to our expectation, PAs and BCs encompassed only 29% (8,046 km2) and 12% (3,185 km2) of suitable areas for dholes, respectively. A vast majority of the areas we deemed suitable for dholes currently remain unprotected, thus making dholes more vulnerable to human persecution and local extermination. We recommend adjusting PA boundaries to fully encompass suitable dhole habitat, and also advocate improved livestock husbandry to reduce dhole related livestock predation and minimize conflict, thereby ensuring its long-term survival in Bhutan.

Cicer turcicum: A New Cicer Species and Its Potential to Improve Chickpea

Genetic resources of the genus Cicer L. are not only limited when compared to other important food legumes and major cereal crops but also, they include several endemic species with endangered status based on the criteria of the International Union for Conservation of Nature. The chief threats to endemic and endangered Cicer species are over-grazing and habitat change in their natural environments driven by climate changes. During a collection mission in east and south-east Anatolia (Turkey), a new Cicer species was discovered, proposed here as C. turcicum Toker, Berger &amp; Gokturk. Here, we describe the morphological characteristics, images, and ecology of the species, and present preliminary evidence of its potential utility for chickpea improvement. C. turcicum is an annual species, endemic to southeast Anatolia and to date has only been located in a single population distant from any other known annual Cicer species. It belongs to section Cicer M. Pop. of the subgenus Pseudononis M. Pop. of the genus Cicer L. (Fabaceae) and on the basis of internal transcribed spacer (ITS) sequence similarity appears to be a sister species of C. reticulatum Ladiz. and C. echinospermum P.H. Davis, both of which are inter-fertile with domestic chickpea (C. arietinum L.). With the addition of C. turcicum, the genus Cicer now comprises 10 annual and 36 perennial species. As a preliminary evaluation of its potential for chickpea improvement two accessions of C. turcicum were field screened for reproductive heat tolerance and seeds were tested for bruchid resistance alongside a representative group of wild and domestic annual Cicer species. C. turcicum expressed the highest heat tolerance and similar bruchid resistance as C. judaicum Boiss. and C. pinnatifidum Juab. &amp; Spach, neither of which are in the primary genepool of domestic chickpea. Given that C. arietinum and C. reticulatum returned the lowest and the second lowest tolerance and resistance scores, C. turcicum may hold much potential for chickpea improvement if its close relatedness supports interspecific hybridization with the cultigen. Crossing experiments are currently underway to explore this question.

Advances in Ethnobotany, Synthetic Phytochemistry and Pharmacology of Endangered Herb Picrorhiza kurroa (Kutki): A comprehensive review (2010-2020)

: Picrorhiza kurroa Royle ex Benth. (Family: Plantaginaceae) is a well-recognized an Ayurvedic herb. It is commonly called “Kutki” or “Kurro” and ‘Indian gentian’. Iridoid glycosides are the plant’s bioactive constituents and accountable for the bitter taste and medicinal properties of the plant. The iridoid glycosides such as picrosides and other active metabolites of the plant exhibited many pharmacological activities like hepatoprotective, antioxidant, anti-inflammatory, anticancer, immunomodulator, anti-ulcerative colitis, antimicrobial etc. This review aims to provide updated information on the ethnobotany, synthetic phytochemistry, pharmacological potential, safety and toxicology of P. kurroa and its active metabolites. Indiscriminate exploitation, ecological destruction of natural habitats, slower plant growth and unawareness regarding cultivation and uprooting of plants has brought kutki as an endangered status. So, various techniques used for the conservation and production of bioactive metabolites from P. kurroa have also been reported. Information on the plant has been collected from Science Direct, Google Scholar, PubMed, Scopus by using ‘Picrorhiza kurroa’, ‘Picroside-‘, ‘Picroside-II’, ‘Picroliv’, ‘Immunomodulator’ keywords. All studies on ethnobotany, phytochemistry and pharmacology of plant from 2010- 2020 were comprised in this review article. The possible directions for the future research have also been outlined in brief in review article.

Comparative genomic analysis of sifakas (Propithecus) reveals selection for folivory and high heterozygosity despite endangered status

Sifakas (genus Propithecus) are critically endangered, large-bodied diurnal lemurs that eat leaf-based diets and show corresponding anatomical and microbial adaptations to folivory. We report on the genome assembly of Coquerel’s sifaka (P. coquereli) and the resequenced genomes of Verreaux’s (P. verreauxi), the golden-crowned (P. tattersalli), and the diademed (P. diadema) sifakas. We find high heterozygosity in all sifakas compared with other primates and endangered mammals. Demographic reconstructions nevertheless suggest declines in effective population size beginning before human arrival on Madagascar. Comparative genomic analyses indicate pervasive accelerated evolution in the ancestral sifaka lineage affecting genes in several complementary pathways relevant to folivory, including nutrient absorption and xenobiotic and fatty acid metabolism. Sifakas show convergent evolution at the level of the pathway, gene family, gene, and amino acid substitution with other folivores. Although sifakas have relatively generalized diets, the physiological challenges of habitual folivory likely led to strong selection.