Rapid behavioral responses of endangered tigers to major roads during COVID-19 lockdown.

Roads pose a major, and growing, challenge for the conservation of endangered species. However, very little is known about how endangered species behaviorally respond to roads and what that means for road mitigation strategies. We used the nation-wide lockdown in Nepal during the COVID-19 pandemic as a natural experiment to investigate how dramatic reductions in traffic volume along the national highway affected movements of two GPS-collared tigers (Panthera tigris)-a globally endangered species. This work is the first systematic research on tigers in Nepal using radiotelemetry or GPS tracking data since the 1980s. We found that the highway more strongly constrained the space use and habitat selection of the male in Parsa National Park than the female in Bardia National Park. Over the entire study period, the female on average crossed 10 times more often per week than the male, and when he was near the highway, he was over 11 times more probable to not cross it than to cross during the day. However, we also found that the cessation of traffic during the pandemic lockdown relaxed tiger avoidance of roads and made the highway more permeable for both animals. They were 2-3 times more probable to cross the highway during the lockdown than before the lockdown. In the month following the lockdown, the space use area of the male tiger tripled in size (160-550 km2), whereas the female's shrunk to half its previous size (33-15 km2). These divergent patterns likely reflect differences between the two parks in their highway traffic volumes and regulations as well as ecological conditions. Our results provide clear evidence that vehicle traffic on major roads impede tiger movements, but also that tigers can respond quickly to reductions in human pressures. We conclude by identifying various actions to mitigate road impacts on tigers and other endangered species.

Non-exploitative human disturbance provides shelter for prey from predator.

Human activities can influence behaviors of predators and prey, as well as predator-prey interactions. Using camera trap data, we investigated whether or to what extent human activities influenced behaviors of predators (tigers and leopards) and prey (sambar deer, spotted deer, wild boar, and barking deer), and predator-prey interactions in the Barandabhar Corridor Forest (BCF), Chitwan District, Nepal. A multispecies occupancy model revealed that the presence of humans altered the conditional occupancy of both prey and predator species. Specifically, the conditional occupancy probability of prey was substantially higher (ψ = 0.91, CI = 0.89-0.92) when humans were present than when humans were absent (ψ = 0.68, CI = 0.54-0.79). The diel activity pattern of most prey species overlapped strongly with humans, whereas predators were generally more active when humans were absent. Finally, the spatiotemporal overlap analysis revealed that human-prey interactions (i.e., the probability that both humans and prey species being present on the same grid at the same hourly period) was ~3 times higher (10.5%, CI = 10.4%-10.6%) compared to spatiotemporal overlap between humans and predators (3.1%, CI = 3.0%-3.2%). Our findings are consistent with the human shield hypothesis and suggest that ungulate prey species may reduce predation risk by using areas with high human activities.

Canine Distemper Virus in Tigers (Panthera tigris) and Leopards (P. pardus) in Nepal.

From wild dogs (Lycaon pictus) in the Serengeti to tigers (Panthera tigris altaica) in the Russian Far East, canine distemper virus (CDV) has been repeatedly identified as a threat to wild carnivores. Between 2020 and 2022, six Indian leopards (P. pardus fusca) presented to Nepali authorities with fatal neurological disease, consistent with CDV. Here, we report the findings of a serosurvey of wild felids from Nepal. A total of 48 serum samples were tested, comprising 28 Bengal tigers (P. t. tigris) and 20 Indian leopards. Neutralizing antibodies were identified in three tigers and six leopards, equating to seroprevalences of 11% (CI: 2.8-29.3%, n = 28) and 30% (CI: 12.8-54.3%, n = 20), respectively. More than one-third of seropositive animals were symptomatic, and three died within a week of being sampled. The predation of domestic dogs (Canis lupus familiaris) has been posited as a potential route of infection. A comparison of existing diet studies revealed that while leopards in Nepal frequently predate on dogs, tigers do not, potentially supporting this hypothesis. However, further work, including molecular analyses, would be needed to confirm this.

Status, distribution and habitat use by Mugger crocodile (Crocodylus palustris) in and around Koshi Tappu Wildlife Reserve, Nepal.

Mugger crocodile is found in various locations of Nepal, ranging from Eastern to Western lowlands, and is also a resident crocodilian of the Koshi Tappu Wildlife Reserve (KTWR). Fewer studies have been carried out on the status and distribution of this crocodilian with limited information on its habitat characteristics. This study sets forth to highlight the population status, distribution and habitat use by mugger crocodile in and around KTWR. Detailed surveys were conducted in the rivers, natural and private ponds of the KTWR in December 2020. Every 500 m along the survey transects, habitat characteristics were recorded in each habitat station as part of the habitat survey. The dependent variables were presence or absence of mugger crocodile in each habitat station. Meanwhile, the independent variables included habitat predictors such as; water depth, slope, elevation, distance to roads, distance to settlements, bank substrate, and human disturbance. Generalized Linear Model under binary logistic regression was employed to test variables associated with presence and absence of muggers for statistical significance. The population survey of the muggers was assessed by day counts either using dugout canoe or walking along transects. During the research period, 35 individuals of muggers were recorded. The probability of mugger sighting augmented with increased slope and depth. 34 muggers (97.14 %) were basking, while 1 (2.85%) were seen swimming out of the total muggers detected. In private and public ponds, 22 muggers were sighted, while 13 were sighted in the main Koshi river, its branches, and the Moriya river. Sixteen muggers (45.71 %) were sighted within the reserve, while the remaining 19 muggers (54.28 %) were sighted in the buffer-zone areas. The probability of mugger sighting varied considerably with regard to slope and water depth among the seven habitat predictors examined. In lieu of prevailing fishing pressure in the river systems of the KTWR and easy availability of prey species for muggers in private fishponds; the muggers are likely use private fishponds more frequently. In order to subdue the muggers inside the KTWR, the reserve authority has a vital role for proper habitat management decisions and regulation of fishing activities.

First report on the molecular detection of Entamoeba bovis from the endangered wild water buffalo (Bubalus arnee) in Nepal.

BACKGROUND: The Asiatic wild water buffalo (Bubalus arnee) is an endangered species that is conserved in the Koshi Tappu Wildlife Reserve (KTWR), Nepal, and was recently translocated to the Chitwan National Park (CNP). Gastrointestinal (GI) parasites are the cause of significant negative health and production impacts on animals worldwide. METHODS: A coprological survey of GI parasites of wild water buffalo was carried out in the CNP in 2020. Fresh dung samples (n = 25) were collected from wild water buffaloes and analysed using sedimentation and flotation techniques for morphological identification of parasite cysts, oocysts and eggs. RESULTS: Nine different GI parasites were recorded of which Entamoeba spp. (20 samples, 80%) were the most common. The presence of Entamoeba spp. was further validated using polymerase chain reaction (PCR) analysis and DNA sequencing. The PCR results were positive for all of the microscopically positive samples, and the species was identified as Entamoeba bovis. Three samples were sequenced and formed a cluster of E. bovis, which was separated from other Entamoeba spp. in phylogenetic analysis. CONCLUSION: This is the first report for molecular detection of E. bovis from wild water buffaloes in Nepal. Future work should focus on the prevalence of such infections in water buffaloes in forest environments.

COVID-19 lockdown frees wildlife to roam but increases poaching threats in Nepal.

To contain transmission of COVID-19, lockdowns or strict restrictions of people's mobility outside their residences were instituted in a majority of countries worldwide, including Nepal, where the first phase of nationwide lockdown was observed from 24 March to 21 July 2020. This sudden halt in human outdoor activities brought positive and negative impacts on forests and wildlife. We undertook a study to learn the impact of the COVID-19 lockdown on wildlife and forests in the protected areas (PAs) of Nepal. Between July and September 2020, data on illegal activities recorded by the staff of PAs and also those reported by media were collected and analyzed. Key informant interviews (KII) were done with the park officers and security personnel by virtual communication (telephone, messenger app, and video call) to collect detailed information and for corroboration. The collected data were categorized into four groups: (a) wildlife killed, (b) wildlife injured, (c) arrest incidents related to forest crime, and (d) arrest incidents related to wildlife crime. Data from the fiscal year 2019-2020 were analyzed, comparing before lockdown and after. Among 20 PAs investigated during the lockdown, the study found substantial increases in wildlife death in two PAs, Banke National Park, and Bardia National Park. Similarly, Chitwan National Park (CNP) and Shivapuri Nagarjun National Park (SNNP) witnessed a rise in wildlife poaching. CNP and SNNP are located close to densely populated cities and also have human settlements in their peripheries. Wildlife was sighted freely roaming inside PAs during the lockdown, presumably because the absence of visitors and human activities during the lockdown decreased disturbance. Thus, the wildlife was enjoying the freedom of movement on the one hand, and on the other hand was threatened by poachers, many of whom were laid off from other activities and were taking advantage of the lapse in security.

Landscape variables affecting the Himalayan red panda Ailurus fulgens occupancy in wet season along the mountains in Nepal.

The Himalayan red panda is an endangered mammal endemic to Eastern Himalayan and South Western China. Data deficiency often hinders understanding of their spatial distribution and habitat use, which is critical for species conservation planning. We used sign surveys covering the entire potential red panda habitat over 22,453 km2 along the mid-hills and high mountains encompassing six conservation complexes in Nepal. To estimate red panda distribution using an occupancy framework, we walked 1,451 km along 446 sampled grid cells out of 4,631 grid cells in the wet season of 2016. We used single-species, single-season models to make inferences regarding covariates influencing detection and occupancy. We estimated the probability of detection and occupancy based on model-averaging techniques and drew predictive maps showing site-specific occupancy estimates. We observed red panda in 213 grid cells and found covariates such as elevation, distance to water sources, and bamboo cover influencing the occupancy. Red panda detection probability [Formula: see text] estimated at 0.70 (0.02). We estimated red panda site occupancy (sampled grid cells) and landscape occupancy (across the potential habitat) [Formula: see text] at 0.48 (0.01) and 0.40 (0.02) respectively. The predictive map shows a site-specific variation in the spatial distribution of this arboreal species along the priority red panda conservation complexes. Data on their spatial distribution may serve as a baseline for future studies and are expected to aid in species conservation planning in priority conservation complexes.

First report of malaria parasites in water buffalo in Nepal.

We present the first molecular-based report on ungulate malaria parasites from water buffalo in Nepal. Fifty-six blood samples were collected from different groups of water buffalo (wild, feral, and domestic) and PCR assays were conducted using Plasmodium spp. cytb specific primers. Two positive cases were detected, one each from feral and domestic individuals. Complete mitochondrial genome sequence (5987 bp) was obtained and examined for nucleotide variations. Sequence analysis revealed identity with type II water buffalo malaria parasites, reported previously, with one A to T nucleotide difference at position 5344. Prevalence, as well as possible economic impacts of water buffalo malaria, should be determined on a wider set of samples from buffalo across Nepal.

Metabolite Profiling and Quantitation of Cucurbitacins in Cucurbitaceae Plants by Liquid Chromatography coupled to Tandem Mass Spectrometry.

Cucurbitaceae is an important plant family because many of its species are consumed as food, and used in herbal medicines, cosmetics, etc. It comprises annual vines and is rich in various bioactive principles which include the cucurbitacins. These steroidal natural products, derived from the triterpene cucurbitane, are mainly the bitter principles of the family Cucurbitaceae. Their biological activities include anti-inflammatory, hepatoprotective, and anti-cancer activities. A total of 10 species belonging to 6 genera of the Cucurbitaceae family along with Cissampelos pareira (Menispermaceae) were included in this study. A comprehensive profiling of certain natural products was developed using HPLC-QTOF-MS/MS analysis and a distribution profile of several major natural products in this family was obtained. A total of 51 natural products were detected in both positive and negative ionization modes, based on accurate masses and fragmentation patterns. Along with this, quantitation of four bioactive cucurbitacins, found in various important plants of the Cucurbitaceae family, was carried out using multiple reaction monitoring (MRM) approach on an ion trap mass spectrometer. Cucurbitacin Q was found to be the most abundant in C. pareira, while Citrullus colocynthis contained all four cucurbitacins in abundant quantities. The developed quantitation method is simple, rapid, and reproducible.

Scientific validation of medicinal plants used by Yakkha community of Chanuwa VDC, Dhankuta, Nepal.

Ethnobotanical knowledge is important among tribal people, but much of the information is empirical due to the lack of scientific validation. The purpose of this study was to document the medicinal plants used by an ethnic group (Yakkha) at Chanuwa VDC of Dhankuta district in Nepal and to validate scientifically in the use of plants based on results of phytochemical, antimicrobial and antioxidant property analyses and available literature reports. Data were collected through interviews of the Yakkha people with the help of a semi-structured questionnaire and the guided field walk method. A total of 30 different medicinal plants were recorded along with their vernacular names (for few plants) used by the Yakkha community's people. Literature review reveals that most of the plant species described herein have also been used in other countries, too. Among 30 plants selected for this study methanol extract of five ethno-medicinal plants viz., Dendrocnide sinuata, Solanum anguivi, Pogostemon cablin, Boehmeria platyphylla and Clerodendrum trichotomum and ethanol extract of C. trichotomum were subjected for antibacterial and antioxidant properties. The antimicrobial activities were measured using the paper disc diffusion method. The antioxidant properties of plants were measured by DPPH and FRAP reduction assay. Among all extracts, ethanol extract of C. trichotomum and methanol extract of B. platyphylla displayed the highest antibacterial and antioxidant activities, respectively.