Ethno-medicinal uses and cultural importance of stingless bees and their hive products in several ethnic communities of Bhutan.

BACKGROUND: Indigenous and non-indigenous people in subtropical and temperate areas of Bhutan share an intricate relationship with stingless bees for diverse purposes including ethno-medicinal uses. Stingless bees hold significant importance in the realms of social, economic, cultural, and spiritual aspects. Bhutan's cultural traditions demonstrate a strong bond with the environment, exemplified by the regular use of honey from stingless bees for remedies such as treating the common cold, cough, and sore throat. METHODS: Ethnographic research was conducted to document the ethno-medicinal uses and cultural importance of stingless in Bhutan. We deployed semi-structured interviews with stingless beekeepers and honey collectors including traditional healers who perform religious rituals for curing and preventing physical and mental illness. RESULTS: We documented 22 different uses of stingless bee honey in food, medicine, veterinary medicine, crafts, beliefs, and religious purposes. The relative cultural importance (RCI) of stingless bees among Bhutan's ethnic communities was assessed through our calculations. It was determined that these bees hold notably greater significance for the Lhotshampa communities compared to other ethnic groups in Bhutan. This finding demonstrates the dependence of Hindu ethnic communities on natural resources in their everyday life. All participant communities largely exploit these bees through destructive extraction practices. They often find the natural nests in nearby forests, transfer them as a log hive to their backyards, and practice traditional meliponiculture. CONCLUSION: The ethnic communities of Bhutan use stingless bees for various purposes and the local knowledge are persistent. However, significant efforts should be made to address the ethno-medicinal, ecological, biological, and commercial perspectives of meliponiculture in Bhutan.

Vitamin D Status among the Population Visiting Jigme Dorji Wangchuck National Referral Hospital, Bhutan.

Objective: To study the vitamin D status among the Bhutanese population visiting the Jigme Dorji Wangchuck National Referral Hospital in Thimphu, Bhutan. Materials and Methods: This is a retrospective descriptive study involving the extraction of data from a hospital database. Records of Bhutanese patients who had taken vitamin D tests in the last two years (2020-2021) were included in the study. Result: A total of 1175 individuals took the vitamin D test during the study period, and the age ranged between 1 day and 94 years. Over 60% of the participants were females. The study found that over 83% of our study population had serum vitamin D levels lower than the normal range/deficient. Around 18% of the participants had severe deficiency. Conclusion: The study found that most participants, including children, had vitamin D deficiency, and the finding was homogenous across gender and age groups. Further studies are required to validate these findings and identify the factors associated with vitamin D deficiency in the population for targeted public health interventions.

Knowledge, attitude and practice on antimicrobial use and antimicrobial resistance among competent persons in the community pharmacies in Bhutan.

Introduction: Since the discovery, antimicrobials have been used to treat variety of infections both in humans and animals caused by microbes. However, with the increasing use, microbes developed resistance to the antimicrobials and many of the antimicrobials became ineffective against certain microbes. Many factors are reported to contribute to the resistance of microbes to antimicrobials. One contributing factor is the misuse and overuse of antibiotics which mainly occur due to the lack of knowledge, careless attitudes, and incorrect practices about use of antibiotics. Methods: This cross-sectional survey study was conducted among the competent persons (CP) in the community pharmacies in Bhutan, with the aim to assess their knowledge, attitude and practice (KAP) on antimicrobial use (AMU) and antimicrobial resistance (AMR). Results: Results from the survey revealed that the competent persons had good level of knowledge about antimicrobial use and antimicrobial resistance. They also had favourable attitude towards antimicrobial resistance and rational use of antimicrobials. Their knowledge and attitude had led to good practices while dispensing antimicrobials from their pharmacies. However, almost all of them had never had any opportunity to take part in activities related to antimicrobial use and antimicrobial resistance that were organized by the public sector. Many of them did not even hear or know about the existence of the policies on use of antimicrobials or on curbing antimicrobial resistance in the country. Conclusion: Involvement of the community pharmacies through trainings and participations in policy making processes is seen as a vital mechanism that can eventually help achieve the goals in the national drive towards reducing antimicrobial resistance.

Current Status and Conservation Management of Farm Animal Genetic Resources in Bhutan.

Farm Animal Genetic Resources (FAnGR) ensures food security and maintains genetic diversity. The efforts to conserve FAnGR are minimal in Bhutan. In the pursuit of increasing livestock productivity, farmers are rearing livestock that narrows the range of genetic diversity. This review attempts to summarize the status of FAnGR and the efforts to conserve them. Some unique livestock breeds in Bhutan are Nublang (cattle breed), Yak, Saphak (pig breed), Yuta (horse breed), Merak-Saktenpa (horse breed), and Belochem (chicken breed). There was a drop in the yak, buffalo, equine, pig, sheep, and goat populations. Both in situ and ex situ conservation measures are in place for some of the breeds and strains (e.g., Nublang and traditional chicken). Conservation efforts are limited to the government, but other individuals, stakeholders and non-government organizations must play an increasing role in preserving genetic diversity. It is pertinent that Bhutan comes up with a policy framework to conserve its indigenous cattle.

A Comprehensive Analysis of Citrus Tristeza Variants of Bhutan and Across the World.

Mandarin orange is economically one of the most important fruit crops in Bhutan. However, in recent years, orange productivity has dropped due to severe infection of citrus tristeza virus (CTV) associated with the gradual decline of citrus orchards. Although the disease incidence has been reported, very limited information is available on genetic variability among the Bhutanese CTV variants. This study used reverse transcription PCR (RT-PCR) to detect CTV in collected field samples and recorded disease incidence up to 71.11% in Bhutan's prominent citrus-growing regions. To elucidate the extent of genetic variabilities among the Bhutanese CTV variants, we targeted four independent genomic regions (5'ORF1a, p25, p23, and p18) and analyzed a total of 64 collected isolates. These genomic regions were amplified and sequenced for further comparative bioinformatics analysis. Comprehensive phylogenetic reconstructions of the GenBank deposited sequences, including the corresponding genomic locations from 53 whole-genome sequences, revealed unexpected and rich diversity among Bhutanese CTV variants. A resistant-breaking (RB) variant was also identified for the first time from the Asian subcontinent. Our analyses unambiguously identified five (T36, T3, T68, VT, and HA16-5) major, well-recognized CTV strains. Bhutanese CTV variants form two additional newly identified distinct clades with higher confidence, B1 and B2, named after Bhutan. The origin of each of these nine clades can be traced back to their root in the north-eastern region of India and Bhutan. Together, our study established a definitive framework for categorizing global CTV variants into their distinctive clades and provided novel insights into multiple genomic region-based genetic diversity assessments, including their pathogenicity status.

Detection and Molecular Characterization of 'Candidatus Liberibacter asiaticus' and Citrus Tristeza Virus Associated with Citrus Decline in Bhutan.

Citrus, mainly mandarin (Citrus reticulata Blanco), is an economically important fruit crop in Bhutan. Despite having favorable agroclimatic conditions for citrus cultivation, the early decline of fruit-bearing orchards coupled with low crop productivity is a major concern among citrus growers. During a recent survey, an association of 'Candidatus Liberibacter asiaticus' (citrus greening) and citrus tristeza virus (CTV), either singly or as mixed infections in declined citrus trees, was recorded in all four major citrus-growing districts (Tsirang, Dagana, Zhemgang, and Sarpang). Using PCR-based diagnosis, a higher incidence of citrus greening (27.45%) and tristeza (70.58%) was observed in symptomatic field samples. Detection and characterization of 'Ca. L. asiaticus' was performed based on the 16S ribosomal DNA, prophage gene, 50S ribosomal rplA-rplJ gene, and tandem repeats of the CLIBASIA_01645 locus. Similarly, the coat protein, p23, and p18 genes were used as genetic markers for the detection and characterization of Bhutanese CTV. The 'Ca. L. asiaticus' isolates from Bhutan segregated into classes II and III based on the CLIBASIA_01645 locus, analogous to Indian isolates from the northeast region and Term-A based on the CLIBASIA_05610 locus. CTV isolates of Bhutan were observed as closely related to the VT strain, which is considered to be the most devastating. To the best of our knowledge, this is the first study on molecular characterization of 'Ca. L. asiaticus' and CTV isolates and their association with citrus decline in Bhutan.

Conservation threats to the endangered golden langur (Trachypithecus geei, Khajuria 1956) in Bhutan.

Threat assessment is critical to species conservation and management planning, because prior identification and assessment of key threats to conservation planning can assist in developing appropriate interventions or strategies. Comprehensive threat assessments are currently lacking for many threatened primates. In this paper, we classify and rank all direct threats to the endangered golden langur (Trachypithecus geei) in Bhutan in order to provide a practical guide to future conservation of the species. Information on threats was based on interviews with local people, discussion with field forestry staff, and social media interaction. We classified threats to golden langur habitats and populations, and ranked them using Miradi™, an analytical software for the adaptive management of conservation projects. We identified five habitat threats: (1) hydropower development, (2) road development, (3) housing development, (4) resource extraction, and (5) agricultural expansion. We also identified seven population threats: (1) electrocution, (2) road kill, (3) road injury, (4) dog kill, (5) retaliatory killing, (6) illegal pet keeping, and (7) hybridization with capped langurs. We rated the overall threat to golden langurs in Bhutan as 'medium'. Hydropower, road, and housing development constituted 'high' impact, while agricultural expansion, resource extraction, electrocution, and road kill had 'medium' impact; the remaining threats had 'low' impact. To immediately mitigate threats to golden langurs, we recommend: (a) installing speed limit signage and speed breakers with strict enforcement of speed limits; (b) installing insulated electric cables and fencing around power transformers; and (c) reducing and restraining domestic dog populations.

Population abundance and distribution of the endangered golden langur (Trachypithecus geei, Khajuria 1956) in Bhutan.

Reliable population estimates are lacking for many South Asian primate species, including the golden langur (Trachypithecus geei), which is endangered and restricted to Bhutan and northeast India. Although well studied in India, few studies exist on this species in Bhutan. In November 2017, we undertook a nationwide survey of golden langurs in Bhutan using double observers along trail-based transects in 17 blocks within its habitat, and modeled its distribution using MaxEnt. A total of 2439 golden langurs in 222 groups were collectively encountered by 17 teams of double observers, from which, an overall population of 2516 ± SE 363 individuals and 236 ± SE 9 groups were estimated. Group sizes varied from 2 to 35 individuals with a mean of 11 ± SD 0.38 individuals. A total of 468 adult males (19%), 924 adult females (38%), 649 juveniles (27%), and 398 infants (16%) were counted. Adult male-to-female sex ratio was 1:1.97 and adult female-to-infant ratio was 1:0.43. We determined 2848 km2 of suitable area for golden langurs in Bhutan and estimated a density of 0.88 individuals/km2. Our population estimate of golden langurs in Bhutan is much lower than the current IUCN estimate of 4000 individuals for Bhutan, necessitating a reassessment of its current conservation status due to threats from road kills, electrocution, and development activities like road construction, hydropower, and electrical transmission lines. We further recommend our refined double-observer survey method to reliably estimate primate populations in rugged terrain.