Mitochondrial DNA analysis of critically endangered Chinese Pangolins (Manis pentadactyla) from Nepal

Chinese Pangolins (Manis pentadactyla) are Critically Endangered and one of the most illegally traded mammals globally. We generated first COI sequences from five individuals of this species from Nepal. BLASTn search of our 600bp sequences at GenBank showed pair-wise identity between 99.17% and 100% to M. pentadactyla. There were three haplotypes and a total of five variable sites among five M. pentadactyla sequences. Neighbor-joining tree revealed that all M. pentadactyla from Nepal clustered into same group further splitting into two sub-groups albeit with low bootstrap value, suggesting potential multiple geographic origins. The K2P distance was 0.3% within group and 0.7% between four sequences from Bhaktapur and Kavrepalanchok districts (Mape2, Mape3, Mape5 and Mape6) and museum sample (Mape10). This study has generated reference samples for M. pentadactyla from Nepal and will be helpful in understanding dynamics of illegal trade of this species and in successful identifi- cation of M. pentadactyla from Nepal even in the absence of intact specimens

Genetic diversity and population structure of Fusarium fujikuroi causing Bakanae, an emerging disease of rice in India

45-52Bakanae caused by Fusarium fujikuroi (Nirenberg), is emerging as a serious threat for rice (Oryza sativa. L.) cultivation in India. In this study, 63 isolates of Fusarium fujikuroi isolated from symptomatic diseased plants were characterized for their morphology, pathogenicity and molecular variability using universal rice primers (URP). Of the 12 URPs used in the study, 6 primers could produce polymorphic fragments in all the isolates. The URP 17R primer was highly polymorphic (100%), whereas, the URP 1F primer produced 75% polymorphic fragments. A dendrogram obtained from the combined analysis of 6 URP primers categorized the isolates into four clusters, where most of the isolates from Punjab and Haryana were clustered separately. Mating type of the population was identified based on MAT-1 and MAT-2 region universal primers for Gibberella fujikuroi. Among the 63 isolates, 18 (28.57%) were identified as MAT-1 and 45 (71.42%) as MAT-2. The effective population number for mating type was 89% of the total population. Since the distribution frequencies of both mating types were not equal in the Indian population of F. fujikuroi, it could conclude that majority of the multiplication of isolates under field conditions was through asexual reproduction. However, the presence of both mating types in F. fujikuroi indicates that the population is also capable of sexual reproduction. Therefore, it is important to develop cultivars with inbuilt resistance to bakanae disease, taking into consideration the factors such as environmental conditions and variability of the pathogen in the area of intended cultivation

Genetic diversity and population structure of Fusarium fujikuroi causing Bakanae, an emerging disease of rice in India

Bakanae caused by Fusarium fujikuroi (Nirenberg), is emerging as a serious threat for rice (Oryza sativa. L.) cultivation in India. In this study, 63 isolates of Fusarium fujikuroi isolated from symptomatic diseased plants were characterized for their morphology, pathogenicity and molecular variability using universal rice primers (URP). Of the 12 URPs used in the study, 6 primers could produce polymorphic fragments in all the isolates. The URP 17R primer was highly polymorphic (100%), whereas, the URP 1F primer produced 75% polymorphic fragments. A dendrogram obtained from the combined analysis of 6 URP primers categorized the isolates into four clusters, where most of the isolates from Punjab and Haryana were clustered separately. Mating type of the population was identified based on MAT-1 and MAT-2 region universal primers for Gibberella fujikuroi. Among the 63 isolates, 18 (28.57%) were identified as MAT-1 and 45 (71.42%) as MAT-2. The effective population number for mating type was 89% of the total population. Since the distribution frequencies of both mating types were not equal in the Indian population of F. fujikuroi, it could conclude that majority of the multiplication of isolates under field conditions was through asexual reproduction. However, the presence of both mating types in F. fujikuroi indicates that the population is also capable of sexual reproduction. Therefore, it is important to develop cultivars with inbuilt resistance to bakanae disease, taking into consideration the factors such as environmental conditions and variability of the pathogen in the area of intended cultivation

Effect of Hydrocortisone on Mortality and Organ Support in Patients With Severe COVID-19: The REMAP-CAP COVID-19 Corticosteroid Domain Randomized Clinical Trial.

Importance: Evidence regarding corticosteroid use for severe coronavirus disease 2019 (COVID-19) is limited. Objective: To determine whether hydrocortisone improves outcome for patients with severe COVID-19. Design, Setting, and Participants: An ongoing adaptive platform trial testing multiple interventions within multiple therapeutic domains, for example, antiviral agents, corticosteroids, or immunoglobulin. Between March 9 and June 17, 2020, 614 adult patients with suspected or confirmed COVID-19 were enrolled and randomized within at least 1 domain following admission to an intensive care unit (ICU) for respiratory or cardiovascular organ support at 121 sites in 8 countries. Of these, 403 were randomized to open-label interventions within the corticosteroid domain. The domain was halted after results from another trial were released. Follow-up ended August 12, 2020. Interventions: The corticosteroid domain randomized participants to a fixed 7-day course of intravenous hydrocortisone (50 mg or 100 mg every 6 hours) (n = 143), a shock-dependent course (50 mg every 6 hours when shock was clinically evident) (n = 152), or no hydrocortisone (n = 108). Main Outcomes and Measures: The primary end point was organ support-free days (days alive and free of ICU-based respiratory or cardiovascular support) within 21 days, where patients who died were assigned -1 day. The primary analysis was a bayesian cumulative logistic model that included all patients enrolled with severe COVID-19, adjusting for age, sex, site, region, time, assignment to interventions within other domains, and domain and intervention eligibility. Superiority was defined as the posterior probability of an odds ratio greater than 1 (threshold for trial conclusion of superiority >99%). Results: After excluding 19 participants who withdrew consent, there were 384 patients (mean age, 60 years; 29% female) randomized to the fixed-dose (n = 137), shock-dependent (n = 146), and no (n = 101) hydrocortisone groups; 379 (99%) completed the study and were included in the analysis. The mean age for the 3 groups ranged between 59.5 and 60.4 years; most patients were male (range, 70.6%-71.5%); mean body mass index ranged between 29.7 and 30.9; and patients receiving mechanical ventilation ranged between 50.0% and 63.5%. For the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively, the median organ support-free days were 0 (IQR, -1 to 15), 0 (IQR, -1 to 13), and 0 (-1 to 11) days (composed of 30%, 26%, and 33% mortality rates and 11.5, 9.5, and 6 median organ support-free days among survivors). The median adjusted odds ratio and bayesian probability of superiority were 1.43 (95% credible interval, 0.91-2.27) and 93% for fixed-dose hydrocortisone, respectively, and were 1.22 (95% credible interval, 0.76-1.94) and 80% for shock-dependent hydrocortisone compared with no hydrocortisone. Serious adverse events were reported in 4 (3%), 5 (3%), and 1 (1%) patients in the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively. Conclusions and Relevance: Among patients with severe COVID-19, treatment with a 7-day fixed-dose course of hydrocortisone or shock-dependent dosing of hydrocortisone, compared with no hydrocortisone, resulted in 93% and 80% probabilities of superiority with regard to the odds of improvement in organ support-free days within 21 days. However, the trial was stopped early and no treatment strategy met prespecified criteria for statistical superiority, precluding definitive conclusions. Trial Registration: ClinicalTrials.gov Identifier: NCT02735707

Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

SummaryBackground Azithromycin has been proposed as a treatment for COVID-19 on the basis of its immunomodulatoryactions. We aimed to evaluate the safety and efficacy of azithromycin in patients admitted to hospital with COVID-19.Methods In this randomised, controlled, open-label, adaptive platform trial (Randomised Evaluation of COVID-19Therapy [RECOVERY]), several possible treatments were compared with usual care in patients admitted to hospitalwith COVID-19 in the UK. The trial is underway at 176 hospitals in the UK. Eligible and consenting patients wererandomly allocated to either usual standard of care alone or usual standard of care plus azithromycin 500 mg once perday by mouth or intravenously for 10 days or until discharge (or allocation to one of the other RECOVERY treatmentgroups). Patients were assigned via web-based simple (unstratified) randomisation with allocation concealment andwere twice as likely to be randomly assigned to usual care than to any of the active treatment groups. Participants andlocal study staff were not masked to the allocated treatment, but all others involved in the trial were masked to theoutcome data during the trial. The primary outcome was 28-day all-cause mortality, assessed in the intention-to-treatpopulation. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936.Findings Between April 7 and Nov 27, 2020, of 16 442 patients enrolled in the RECOVERY trial, 9433 (57%) wereeligible and 7763 were included in the assessment of azithromycin. The mean age of these study participants was65·3 years (SD 15·7) and approximately a third were women (2944 [38%] of 7763). 2582 patients were randomlyallocated to receive azithromycin and 5181 patients were randomly allocated to usual care alone. Overall,561 (22%) patients allocated to azithromycin and 1162 (22%) patients allocated to usual care died within 28 days(rate ratio 0·97, 95% CI 0·87–1·07; p=0·50). No significant difference was seen in duration of hospital stay (median10 days [IQR 5 to >28] vs 11 days [5 to >28]) or the proportion of patients discharged from hospital alive within 28 days(rate ratio 1·04, 95% CI 0·98–1·10; p=0·19). Among those not on invasive mechanical ventilation at baseline, nosignificant difference was seen in the proportion meeting the composite endpoint of invasive mechanical ventilationor death (risk ratio 0·95, 95% CI 0·87–1·03; p=0·24).Interpretation In patients admitted to hospital with COVID-19, azithromycin did not improve survival or otherprespecified clinical outcomes. Azithromycin use in patients admitted to hospital with COVID-19 should be restrictedto patients in whom there is a clear antimicrobial indication

Tocilizumab in patients admitted to hospital with COVID-19 (RECOVERY): a randomised, controlled, open-label, platform trial

Background: In this study, we aimed to evaluate the effects of tocilizumab in adult patients admitted to hospital with COVID-19 with both hypoxia and systemic inflammation. Methods: This randomised, controlled, open-label, platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]), is assessing several possible treatments in patients hospitalised with COVID-19 in the UK. Those trial participants with hypoxia (oxygen saturation <92% on air or requiring oxygen therapy) and evidence of systemic inflammation (C-reactive protein ≥75 mg/L) were eligible for random assignment in a 1:1 ratio to usual standard of care alone versus usual standard of care plus tocilizumab at a dose of 400 mg–800 mg (depending on weight) given intravenously. A second dose could be given 12–24 h later if the patient's condition had not improved. The primary outcome was 28-day mortality, assessed in the intention-to-treat population. The trial is registered with ISRCTN (50189673) and ClinicalTrials.gov (NCT04381936). Findings: Between April 23, 2020, and Jan 24, 2021, 4116 adults of 21 550 patients enrolled into the RECOVERY trial were included in the assessment of tocilizumab, including 3385 (82%) patients receiving systemic corticosteroids. Overall, 621 (31%) of the 2022 patients allocated tocilizumab and 729 (35%) of the 2094 patients allocated to usual care died within 28 days (rate ratio 0·85; 95% CI 0·76–0·94; p=0·0028). Consistent results were seen in all prespecified subgroups of patients, including those receiving systemic corticosteroids. Patients allocated to tocilizumab were more likely to be discharged from hospital within 28 days (57% vs 50%; rate ratio 1·22; 1·12–1·33; p<0·0001). Among those not receiving invasive mechanical ventilation at baseline, patients allocated tocilizumab were less likely to reach the composite endpoint of invasive mechanical ventilation or death (35% vs 42%; risk ratio 0·84; 95% CI 0·77–0·92; p<0·0001). Interpretation: In hospitalised COVID-19 patients with hypoxia and systemic inflammation, tocilizumab improved survival and other clinical outcomes. These benefits were seen regardless of the amount of respiratory support and were additional to the benefits of systemic corticosteroids. Funding: UK Research and Innovation (Medical Research Council) and National Institute of Health Research

Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

Background: Many patients with COVID-19 have been treated with plasma containing anti-SARS-CoV-2 antibodies. We aimed to evaluate the safety and efficacy of convalescent plasma therapy in patients admitted to hospital with COVID-19. Methods: This randomised, controlled, open-label, platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]) is assessing several possible treatments in patients hospitalised with COVID-19 in the UK. The trial is underway at 177 NHS hospitals from across the UK. Eligible and consenting patients were randomly assigned (1:1) to receive either usual care alone (usual care group) or usual care plus high-titre convalescent plasma (convalescent plasma group). The primary outcome was 28-day mortality, analysed on an intention-to-treat basis. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936. Findings: Between May 28, 2020, and Jan 15, 2021, 11558 (71%) of 16287 patients enrolled in RECOVERY were eligible to receive convalescent plasma and were assigned to either the convalescent plasma group or the usual care group. There was no significant difference in 28-day mortality between the two groups: 1399 (24%) of 5795 patients in the convalescent plasma group and 1408 (24%) of 5763 patients in the usual care group died within 28 days (rate ratio 1·00, 95% CI 0·93–1·07; p=0·95). The 28-day mortality rate ratio was similar in all prespecified subgroups of patients, including in those patients without detectable SARS-CoV-2 antibodies at randomisation. Allocation to convalescent plasma had no significant effect on the proportion of patients discharged from hospital within 28 days (3832 [66%] patients in the convalescent plasma group vs 3822 [66%] patients in the usual care group; rate ratio 0·99, 95% CI 0·94–1·03; p=0·57). Among those not on invasive mechanical ventilation at randomisation, there was no significant difference in the proportion of patients meeting the composite endpoint of progression to invasive mechanical ventilation or death (1568 [29%] of 5493 patients in the convalescent plasma group vs 1568 [29%] of 5448 patients in the usual care group; rate ratio 0·99, 95% CI 0·93–1·05; p=0·79). Interpretation: In patients hospitalised with COVID-19, high-titre convalescent plasma did not improve survival or other prespecified clinical outcomes. Funding: UK Research and Innovation (Medical Research Council) and National Institute of Health Research

Population changes in Gharial (Gavialis gangeticus) vary spatially in Chitwan National Park, Nepal

Gharials, large crocodilians found only in South Asia, are widely seen as a flagship species for river conservationin Nepal, especially in Chitwan National Park, where a headstart program has supplemented the populationsince 1981. The population has shown signs of recovery only in the last decade, so continued monitoring of populationtrends is vital for conservation. We conducted annual winter population surveys for gharial in Chitwan between 2017and 2022, during which we also characterized riverbank substrate availability and basking preferences. We documentedpotential threats to the species in Chitwan throughout the year. Overall, we counted an increasing number of Gharialsin Chitwan; however mixed-effects modelling of Gharial encounter rate showed that increasing encounters rates are notevenly distributed throughout available habitat, with some river stretches having stable or decreasing trends. Encounterrates on the Rapti River increased in all transects, compared to more variable results on the Narayani River, likelyattributable to higher levels of human disturbance and the impact of captivity on habitat selection. Fewer Gharials wereseen in transects with high levels of disturbance due to sand mining and the extraction of river substrates, highlightingthis threat as a major concern. Regular reports of bycatch in illegal gillnets was the major observed source of mortality.A lack of an increasing population trend in the stretch above a large barrage suggests that recruitment is minimal inthis area, and the dam likely has a negative impact on upstream Gharial recruitment. We cautiously suggest that theChitwan population is recovering, but that recovery is hampered by threats, especially substrate extraction, illegal gillnetfishing, and river fragmentation by a dam

Not Available

Not AvailableSclerotinia stem rot caused by Sclerotinia sclerotiorum (Lib.) de Bary was observed for the first time on pigeonpea in Pusa, Bihar (India) during the years 2012–2014. Typical symptoms of the disease included blighting of twigs, stem lesions, and development of cottony white mycelium and numerous large sclerotia on diseased plant parts. The fungus was identified on the basis of cultural and morphological characteristics, and its pathogenicity was also established. The internal transcribed spacer (ITS) region of pathogen was amplified with primers ITS1 and ITS4, which revealed 100% query coverage along with 99% identity with S. sclerotiorum recovered from different hosts. The highest disease incidence was recorded on cultivar ‘ICPL151’, whereas cultivars ‘MAL13’ and ‘Kudrat’ were observed to have significantly less disease under natural conditionsNot Availabl

Understanding the perception of buffer zone communities to gharial (Gavialis gangeticus) conservation in Chitwan National Park, Nepal

The gharial (Gavialis gangeticus) is a Critically Endangered species found in Nepal and India. In Nepal, gharials are distributed in Chitwan and Bardia National Parks. Gharials are highly threatened due to various and severe anthropogenic activities. In this study, we assess the attitude, perception and willingness of a range of stakeholders living around Chitwan National Park to participate in gharial conservation. We conducted semi-structured questionnaire surveys with open ended questions. We interviewed national park officials (n = 8), local community group members (n = 54), hoteliers (n = 12), conservation partners (n = 5), local elected representatives (n = 38), fishing community members (n = 14), fishing net sellers (n = 15) and students (n = 9). Our study revealed that respondents are familiar with existing conservation awareness activities and expressed their willingness to participate in gharial conservation activities which can be capitalized on for future conservation initiatives for gharials. Further, we suggest to concentrate efforts particularly on local representatives, fishing net sellers and river dependent communities. These efforts include community level gharial knowledge sharing and interaction program, school teaching program, participating community in gharial conservation and its habitat management intervention and livelihood programs