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National Mission on Himalayan Studies (NMHS) HIMALAYAN RESEARCH FELLOWSHIP (PRO FORMA FOR THE ANNUAL PROGRESS REPORT) [Reporting Period: from 1st April, 2016 to 30th March, 2017]

Kindly fill the NMHS Fellowship Annual Progress Report segregated into the following 7 segments, as applicable to the NMHS Fellowship nature and outcomes.

1. Fellowship Grant Information and Other Details

2. Fellowship Description at Himalayan Research Associates (H-RAs) Level

3. Fellowship Description at Himalayan Junior Research Associates (H-JRFs) Level

4. Fellowship Description at Institutional/ University Level

5. Fellowship Concluding Remarks/ Annual Summary

6. Specific Research Question(s) Addressed with Succinct Answer(s)

7. Any other information

Please let us know in case of any query at: [email protected]

PRO FORMA NMHS-Fellowship Annual Progress Report (APR)

1. Fellowship Grant Information and Other Details

NMHS Fellowship Grant ID: HSF2015-16_I003

Name of the Institution/ University: Zoological Survey of India, Kolkata

No. of Himalayan Research/Project Associates: 03

No. of Himalayan Junior Research/Project Fellows: 10

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2. Fellowship Description at H-RA Level

Himalayan Research Associates (H-RAs)

H-RAs Profile Description:

Name of the PI and S. No. Name of RA Date of Joining Research Title Qualification Designation

1. Dr. Abesh 03.06.2016 Lepidoptera (Insecta) as Dr. Kailash Ph.D. Kumar Sanyal Potential Indicator-Taxa Chandra, Director for Tracking Climate Change in the Indian Himalayan Landscape

2. Dr. Arockia 13.10.2016 “ Dr. Kailash Ph.D. Lenin Chandra, Director

3. Dr. John Caleb 01.03.2017 “ Dr. Kailash Ph.D. Chandra, Director

Progress Brief (to be filled for each H-RA in separate row):

Research Research/ RA No. Addressed Deliverables Achievements Objective(s) Experimental Work*

1.  Number of Long-Term  Identification of sites for  Initial LTEM identified at HNP (J & [Enclose descriptive file – Ecological/ LTEM. K), GHNP (Himachal Pradesh), AWLS max. 250 words] Environmental  Habitat-suitability mapping of (Uttarakhand), SNP/NVNP (West Monitoring (LTEM) sites threatened Himalayan Bengal) and DDBR (Arunachal establishment. butterflies. Pradesh).  Status & distribution of  Conservation frameworks  Synthesis of secondary information threatened Apollo and development for Himalayan on Himalayan lepidoptera including other Parnassini Lepidoptera. Apollos. butterflies.  Historical sites for repeat monitoring identified.  Identification of collected moths. 2  Monitoring surveys in  Predictive species  Species validation from secondary [Enclose descriptive file – historical collection distribution modeling for literature. Reconnaissance survey at max. 250 words] localities. future climatic scenario. SNP/NVNP (West Bengal).  Climate-Envelope  Landscape level  Sample preparation for DNA modeling & Distribution conservation approach of barcoding. mapping for responses Himalayan Lepidoptera  Identification of collected to changing climate. through Indicator taxa and Lepidoptera. awareness generation. 3.  Molecular Phylogenetic  A barcode database  Curation and alignment of [Enclose descriptive file – Work through DNA generation for future generated sequences of Lepidoptera max. 250 words] Barcoding to resolve molecular and phylogenetic specimens with the representative species complexes researches on Himalayan barcode sequences from global Lepidoptera database for similarity search.

 The developed sequences were submitted to BOLD database under the project ‘Lepidoptera of Indian Himalayas’ for acquiring the BOLD- IDs.

*Experimental work giving full details (in separate sheet, within 300 words) of experimental set up, methods adopted, data collected supported by necessary table, charts, diagrams & photographs. Note: Data, table and figures may be attached as separate source file (.docx, .xls, jpg, .jpeg, .png, .shp, etc. ): Descriptive file and Compiled information on the progress of the project under NMHS fellowship has been attached.

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3. Fellowship Description at H-JRF Level Himalayan Junior Research Project Fellows (H-JRFs)

H-JRFs Profile Description: S. No. Name of JRF Date of Joining Name of the PI Qualification

1. Mohd. Ali 29.09.2016 Dr. Kailash Chandra M.Sc.

2. Kaushik Mallick 06.05.2016 Dr. Kailash Chandra M.Sc.

3. Uttaran Bandyopadhyay 14.05.2016 Dr. Kailash Chandra M.Sc.

4. Rahul Ranjan 24.02.2017 Dr. Kailash Chandra M.Sc.

5. Kamalika Bhattacharyya 14.05.2016 Dr. Kailash Chandra M.Sc.

6. Subrata Gayen 14.05.2016 Dr. Kailash Chandra M.Sc.

7. Gaurab Nandi Das 14.05.2016 Dr. Kailash Chandra M.Sc.

8. Sumantika Chatterjee 03.05.2016 Dr. Kailash Chandra M.Sc.

9. Angshuman Raha 03.05.2016 Dr. Kailash Chandra M.Sc.

10. Rohit Kumar Jaiswal 15.05.2016 Dr. Kailash Chandra M.Sc.

Progress Brief (to be filled for each JRF in separate row): Research/ JRF Research Objectives Deliverable Achievements Experimental No. Work*

1.  To investigate the differential  Generation of  Field survey at HNP (J & K). [Enclose processes influencing the scientific evidences in  Processing of collected Lepidoptera from HNP. descriptive file – distribution pattern of the form of reports  Secondary data entry for Apollos for species max. 250 words] on Lepidopteran Lepidoptera assemblages distribution modelling. diversity of HNP.  Registration and course work done for Ph.D. (moths and butterflies) of  Robust data sets. programme at Saurashtra University on the topic HNP.  Thesis on the subject ‘Assessing Diversity and Distribution of  Generation of robust datasets area. Lepidoptera focusing on Apollos as Flagship for generated through ecological Biodiversity Conservation in Trans-Himalayan monitoring at habitat level Landscape of Ladakh’. 2. •To investigate the differential  Generation of  Field survey at GHNP (Himachal Pradesh). [Enclose processes influencing the scientific evidences in  Processing of collected Lepidoptera from GHNP. descriptive file – Lepidoptera diversity of GHNP the form of reports  Morpho-taxonomy including genitalia dissection max. 250 words] •Generation of robust datasets on Lepidopteran of collected Geometrid moths generated through ecological diversity of GHNP.  Species validation from secondary literature. monitoring at habitat level  Robust data sets.  Registration and course work done for Ph.D. Generation of scientific  Thesis on the subject programme at Saurashtra University on the topic evidences area. ‘Diversity and Distributional Pattern of Family Geometridae in GHNP Conservation Area’. 3. •To investigate how diverse  Generation of  Field survey at AWLS (Uttarakhand). [Enclose are the Lepidopteran scientific evidences  Processing of collected Lepidoptera from AWLS. descriptive file – assemblages in AWLS in the form of  Morpho-taxonomy including genitalia dissection of max. 250 words] •Generation of robust datasets reports on collected Noctuid moths generated through ecological Lepidopteran  Preparation of Indian Notodontidae Checklist. monitoring at habitat level diversity of AWLS.  Data entry from secondary literature.  Robust data sets.  Registration and course work done for Ph.D.  Thesis on the programme at Saurashtra University on the topic subject area. ‘Diversity and Distributional Pattern of Moths with Special Emphasis on Family Noctuidae in AWLS, Uttarakhand’. 4.  To investigate the differential  Generation of  Processing of collected Lepidoptera for DNA [Enclose processes influencing the scientific evidences sequencing. descriptive file – Lepidoptera diversity of KBS in the form of •Species identification of collected samples from max. 250 words]  Generation of robust datasets reports on Central Himalaya. generated through ecological Lepidopteran monitoring at habitat level diversity of AWLS.

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 Robust data sets.  Thesis on the subject area 5. •To investigate the differential • Generation of  Field survey at SNP/NVNP (West Bengal). [Enclose processes influencing the scientific evidences in  Processing of collected Lepidoptera from descriptive file – Lepidoptera diversity of SNP the form of reports on Central Himalaya. max. 250 words] •Generation of robust datasets Lepidopteran diversity  Species validation and entry from secondary generated through ecological of SNP literature. monitoring at habitat level • Robust data sets  Species identification of Central Himalaya • Thesis on the subject collections. area  Registration and course work done for Ph.D. programme at Saurashtra University on the topic ‘Moth Assemblages along Altitudinal Gradient in SNP and NVNP- An Ecological and Molecular Approach Emphasizing on Noctuidae’ 6.  To investigate the  Generation of  Field survey at DDBR (Arunachal Pradesh). [Enclose differential processes scientific  Processing of collected Lepidoptera from descriptive file – influencing the evidences in the Eastern Himalaya. max. 250 words] distribution pattern of form of reports  Compilation of secondary data on Himalayan Lepidoptera assemblages on Lepidopteran butterfly. (moths and butterflies) diversity of DDBR  Validation of Himalayan Pyralidae from  Generation of robust  Robust data sets Secondary literature. datasets generated Thesis on the  Ongoing field sampling at DDBR from 22nd through ecological subject area March 2017 monitoring at habitat level 7.  To investigate the  Generation of  Butterfly sampling at Central Himalaya and [Enclose differential processes scientific DDBR (Arunachal Pradesh). descriptive file – influencing the evidences in the  Synthesis of Himalayan butterfly from max. 250 words] distribution pattern of form of reports secondary literature. Lepidoptera assemblages on Lepidopteran  Processing of collected Lepidoptera from (moths and butterflies) diversity of Eastern Himalaya. in DDBR DDBR  Identification of collected butterfly samples.  Generation of robust  Robust data  Ongoing field sampling at DDBR from 22nd datasets generated sets March 2017. through ecological  Thesis on the monitoring at habitat subject area level 8.  Study of Molecular  Report on status  DNA isolation using organic phase extraction [Enclose Taxonomy of molecular method/silica column kit method/and descriptive file – taxonomy magnetic bead based protocol. max. 250 words]  Amplification of isolated DNA samples.  Analysis of forward and reverse chromatograms to obtain the consensus sequences. 9.  Morphology-based  Report on status  Field survey at GHNP (Himachal Pradesh). [Enclose Taxonomy of morpho-  Morphology based identification of collected descriptive file – taxonomy & moth samples. max. 250 words] generation of  Genitalia based identification of Noctuidae, identification Notodontidae, Geometridae and Erebidae. keys  Validation of Himalayan Lepidoptera from Secondary literature.  Synthesis of secondary data on Himalayan Lepidoptera. 10.  Modelling on GIS-  Robust database  Boundary demarcation of PAs included in the [Enclose framework and Data on species/ project. descriptive file – Compilation habitats  Preparation of Digital Elevation Map (DEM), max. 250 words]  Comparison of robust  Strategy for Contour and Aspect maps of PAs. datasets generated conservation  For NDVI, Landsat 8 and MODIS data procured through ecological from USGS. monitoring  Climatic data downloaded from WorldClim.  Workshop on MAXENT attended at Indian Statistical Institute, Kolkata.  Field survey at SNP/NVNP, GHNP, AWLS and DDBR for ground truthing. *Experimental work giving full details (in separate sheet, within 300 words) of experimental set up, methods adopted, data collected supported by necessary table, charts, diagrams & photographs. Note: Data, table and figures may be attached as separate source file (.docx, .xls, jpg, .jpeg, .png, .shp, etc. ). Descriptive file and Compiled information on the progress of the project under NMHS fellowship has been attached.

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4. Fellowship Description at Institutional/ University Level

Annual Deliverables/ Outputs (during the reporting year)

S. No. Deliverables/ Parameters No. Description 1. No. of Research Publications (monograph/ articles/ peer-reviewed 02 [Enclose file for description – max. 250 words] articles): 2. No. of Data Sets generated: 03 [Enclose file for description – max. 250 words] 3. No. of Conferences/ Workshops 01 [Enclose file for description – max. 250 words] attended: 4. No. of Sites/ Study Area Covered: 06 [Enclose file for description – max. 250 words] 5. No. of Best Practices suitable for IHR: Need further investigation 6. New Observations/ Innovations 10 [Enclose file for description – max. 250 words] Descriptive file and Compiled information on the progress of the project under NMHS fellowship has been attached.

5. Fellowship Concluding Remarks/ Annual Summary

Conclusions summarizing the achievements and indication of remaining work (within 300 words):

 Recruitment and orientation of 03 Ras and 10 JRFs have been achieved. Literature review on Butterflies and Moths of Indian Himalayan Region has been completed with documentation of 1187subspecies/985 species of butterflies and 3533 species of moths from 6 Himalayan states. Study sites selection and initial reconnaissance surveys were undertaken in all the proposed landscape, except KBR in Sikkim, where work can’t be initiated as permission from Sikkim Forest Department has still not be sanctioned. Field sampling has been done in 12 field sessions in 5 Himalayan states with recording of 4863 number of specimens among which 1464 specimens have been identified following standard taxonomic procedure. Laboratory works were undertaken for morpho and molecular taxonomy which resulted in total 100 nos. of plates depicting genitalic structure of moths and 368 DNA sequences generated which were submitted to BOLD.  The remaining works include: 1. Extensive sampling in all the sites selected for LTEMs which will include light trapping, butterfly transects, vegetation sampling and recording of all the biotic-abiotic parameters influencing the Lepidoptera diversity and distribution. For next two years, sites will be sampled in season-wise covering three main seasons, viz. Pre-Monsoon, Monsoon and Post-Monsoon. 2. Monitoring surveys in Historical collection localities will be undertaken in 12 sites identified from which significant past records on Lepidoptera have been documented. Species distribution map will be generated from primary as well as secondary data, and changes in recorded altitude preferences for individual species will be compared, which will ultimately give indication whether Himalayan Lepidoptera is undergoing any significant changes in distribution and range size due to global climatic warming. 3. Extensive sampling in Trans-Himalayan section of Jammu & Kashmir will be undertaken to record species of Apollo butterflies and their host-plant and biotic parameters which may govern their distribution. Moreover, questionnaire surveys will be undertaken among local people residing in Apollo distribution range to assess the perception of local communities about their potential as Flagship species for biodiversity conservation. Moreover, this data will be used to prepare “Habitat Suitability Mapping” and conservation plan for their long- term survival. 4. DNA barcoding of all the collected Lepidoptera samples will be continued to resolve cryptic species complex and preparing barcode database for future molecular and phylogenetic

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researches.

6. Specific Research Question(s) Addressed with Succinct Answer(s)

S. No. Research Questions Addressed Succinct Answers (within 150-200 words) 1. How diverse are the Lepidopteran Literature review revealed more than 1000 and assemblages in different Himalayan 3500 species of moths and butterflies, Biogeographic Provinces? respectively, from IHR. Butterfly diversity is maximum in Arunachal Pradesh followed by that of Sikkim. While, maximum moth species were recorded from Sikkim followed by Uttarakhand. From field sampling of the present study 1464 number of specimens have so far been been identified which comprises 507 morphospecies under 265 genera (details provided in Table 7). The present study shows North-Western and Western Himalaya is the most diverse biogeographic province with 245 species under 158 genera followed by Central and Eastern Himalaya including 158 and 104 species, respectively. This is preliminary observation, more sampling are required for clear understanding of Lepidopteran assemblage pattern across Himalayan biogeographic provinces. 2. What are the major biotic-abiotic Different biotic and abiotic factors are being factors including anthropogenic collected during field sampling of the present disturbances shaping the distribution study. These include weather parameters, such pattern of Lepidopteran assemblages in as temperature and relative humidity at every extreme Himalayan Biogeographic one hour interval during sampling, wind speed, Provinces? cloud cover etc., ambient light, vegetation parameters, such as canopy cover, tree and shrub density, herb cover, litter cover, dominant tree and shrub species etc. and disturbance parameters, such as logging percentage, grazing intensity, fire sign, distance of human habitation etc. Further sampling are required to understand the effects of these variables as well as to identify relevant variables in shaping the distribution pattern of Lepidopteran assemblages in IHR. 3. Which are the altitudinally restricted, Primary field sampling indicated several species thus climatically vulnerable species from family Geometridae, such as Photoscotosia assemblages and what are the non- amplicata and P. multilinea, Opisthograptis climatic threats to their long term tridentifera and O. sulphurea, Abraxas sp., from survival? family Noctuidae, such as Diphtherocome pallida and D. fasciata, Hermonassa sp., Xestia sp., Valeriodes sp., Phlogophora sp. from family Erebidae, like Spilosoma leopardine and from Bombycidae, such as Mustilia sphingiformes etc. are high altitude restricted species which are only found in the altitude range from 3200-4000 m. These taxa will be monitored, particularly in the present study to see altitudinal shifts in their distribution pattern, change in abundance and

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phenology and thus, can be used as potential indicator for climate change. Observation from extensive field sampling of the present study will identify more such altitudinally restricted species whose predictive distribution modelling will provide new insight on the effect of ongoing climate change scenario. 4 Are there any significant changes in More than 300 historical sites from six different species composition and species states of Indian Himalaya were identified from altitudinal ranges in the historically secondary literature. Species assemblages prominent collection localities in recorded from these sites will be compared with Himalaya? the data of present study for investigating climate change effects on Lepidoptera and associated taxa. Secondary information revealed maximum historical sites from Uttarakhand followed by West Bengal and Sikkim. 5 What are the major directions towards Further explorations of the historical sites are which species distribution range is required to answer this question. shifting due to climatic modifications over last one century? 6 Which Himalayan species/species Preliminary investigation identified several complexes will be facing extinction risks species from Geometridae and Noctuidae as and where will be the suitable habitats high altitude restricted species. These species for their survival in future climatic are facing extinction risk due to ongoing climatic scenario? perturbations. More sampling are required to generate predictive distribution modeling of these and even more target species which will be identified in due course of the present study. 7 What is the population status of Geo-referenced distribution records of 18 threatened species of Apollos and species of Apollos from published literature have other butterfly species of subfamily been entered in specific MS Excel data sheet. Parnassinae, Family Papilionidae? During field work of the present study two species of Apollo have been encountered till date. To comment on the population status of Apollos more extensive samplings are required. 8 Which ecological factors define their Different climatic and habitat variables are being optimum habitat conditioning? recorded for understanding the factors determining Apollo distribution. More sampling are required for better interpretation of data. 9 Under projected future climatic These questions will be answered after scenario, how and where these generating robust dataset of Apollo and other threatened butterflies will be Parnassinae distribution across Indian Himalayan surviving? What will be the key Region from past studies and present field management steps towards their long sampling. term conservation?

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Table 2: Fellowship Description at H-RA Level

Dr. Abesh Kumar Sanyal, RA

Initial Gradsects (altitudinal transects covering a complete altitudinal gradient) were established in HNP, GHNP, AWLS, SNP and DDBR, which will be monitored and surveyed repeatedly (LTEMs) for next two year’s field sampling. The gradsects were chosen so as to cover maximum possible habitat- heterogeneity in each landscape covering all possible habitat types, so that the Lepidopteran fauna characteristic for each habitat type can be sampled. In each gradient sampling stations were marked at every 300m vertical distance. Sampling points were chosen in such a way that it sits within a homogenous vegetation cover representing the characteristic vegetation type of that altitudinal band and light trapping from there shouldn’t attract species from adjacent habitat category. Initial sampling in each sampling stations recorded considerable amount of Lepidopteran fauna (details provided in Table 1). All the samples were processed and curated properly and a significant portion of them were identified (Table 7) following standard taxonomic operation.

The secondary records on moths from Indian Himalayan Region was compiled and analysed against major biotic-abiotic factors. To prepare the primary dataset, material examined or specimen details of species wherever available were extracted and geo-referenced. Sampling points were transformed into richness grids which were then overlaid on altitude, annual precipitation and vegetation layers separately to show moth richness patterns along the IHR. The status of secondary information and potential sites for repeating monitoring surveys are provided in Table 9, 10 & 11.

Dr. Arockia Lenin, RA

Preliminary field surveys for Lepidoptera inventory undertaken during November-December, 2016 in Singalila NP & Neora Valley NP, Central Himalaya. The survey resulted in around 200 moth specimens (details given in Table 5), which were stretched, labelled and curated for further taxonomic studies. Specimens from Central Himalaya and Askot Wildlife Sanctuary were identified following standard taxonomic procedure.

Species validation of huge secondary record of moths from entire Himalayan landscape were undertaken through LEPINDEX (www.nhm.ac.uk/our-science/data/lepindex) and PROJECT LIFE (www.nic.funet.fi/pub/sci/bio/life/insecta/lepidoptera) database. This resulted in generation of updated species information with type locality, original genus, current status, rank, higher classification and geographic distribution.

About 403 identified Lepidoptera samples have been given NMH IDs for DNA leg extraction, and about 226 identified samples have been chopped and processed for DNA isolation by using standard procedure. In addition, The Barcode of Life Data (BOLD) systems details like voucher info (specimen id, field specimen id, museum id, collection code, institution storing), taxonomy (Phylum, class, order, Family, subfamily, genus, species, identifier, identifier method, identifier institution, identification method, tamonomy notes, common name), collection data (sample id, collectors, collection date, Country, State, Region, Sector, latitude, longitude, elevation etc.,) and specimen data (sex, reproduction, life stage, extra info, notes, voucher status and specimen metadata extended fields, etc.) of collected specimens from ASKOT, GHNP, CENTRAL HIMALAYA and DDBR landscapes have been updated.

Dr. John Caleb, RA

The data from the morphological identifiers and geographical coordinates were updated in a common data file. The samples were then processed for DNA extraction. The legs were removed and stored in -20˚C until further process. The stored samples were then processed for DNA extraction. PCR amplification of the approximately 500 base pair (bp) in the mtDNA marker, COI gene was performed. The PCR product was purified by using QIAquickR Gel extraction kit. The bidirectional sequencing of the purified PCR product was carried out by using BigDye® Terminator Cycle Sequencing Kit (v3.1) on 3730 DNA Analyzer (Applied BioSystems) in the in-house sequencing facility of Zoological Survey of India, Kolkata. The generated sequences of 450 Lepidoptera specimens were curated and aligned with the representative barcode sequences from the global database for similarity search. Global database on Lepidoptera has been retrieved from Barcode of Life Data Systems (BOLD). The developed sequences were submitted to BOLD database under the project ‘Lepidoptera of Indian Himalayas’ for acquiring the unique BOLD-IDs for accession. The following batches of specimens identified morphologically are being processed in the similar manner to obtain the barcodes. Barcode database of each specimen has been generated for phylogenetic analysis. These sequences would be used to build phylogenetic trees by specialized software to construct the best fit model. This is also used to compare genetic distinctness of cryptic species complex of Lepidoptera.

Table 3: Fellowship Description at H-JRF Level

Mohd. Ali, JRF

Reconnaissance survey was conducted in Hemis National Park in the vicinities of Hemis Village, Sumdo, Nagdang Village, Ngdang Pho-I, Nagdand Pho-II and Nagdang Pho-II within the Hemis National Park (details provided in Page 2, Table 2). Collected specimens were sorted according to families, stretched and tagged for further taxonomic study. Majority of the specimens were from family Noctuidae comprising 18 morpho-species. One species of Apollo was sighted during the fieldwork and local villagers were approached for the questionnaire about different issues pertaining to Lepidoptera diversity and agriculture practice to make an effective conservation plan.

Secondary records of Apollos from Trans, North-Western and Western Himalaya were compiled and entered in specific Excel format for preparing baseline database for species distribution modelling (Details provided in Table 12). Thus, a total of 18 Apollo species were recorded from secondary literatures. Moreover, more the 40 species of host plants of Apollos have been identified through secondary literature to improve the efficiency of next survey.

Selected for the Ph.D. program at Saurashtra University, Gujarat and completed course work (from 02.01.2017 to 23.03.2017) at Wildlife Institute of India. Ph.D. synopsis on the topic of “Assessing Diversity & Distribution of Lepidoptera focusing on Apollos (Papilionidae: Parnassini) as Flagship for Biodiversity Conservation in Trans-Himalayan Landscape of Ladakh” has been prepared and submitted to Saurashtra University.

Kaushik Mallick, JRF

Preliminary surveys for selecting sampling sites and making primary inventory were undertaken during August-September, 2016 and November-December, 2016 in Great Himalayan National Park, Himachal Pradesh (details provided in Table 4). Specimens were collected from different habitats installing light traps running for 4 hours, in two sessions at a particular site for two consecutive nights, e.g. first night from 8 pm to 12’O clock midnight and second night from 12’o clock midnight to 4 am, to ensure recording all the moths flying in different quarters of night. For setting light traps, Mercury bulb (160W) or a solar powered lantern equivalent to 80 Watt White Actinic bulb and gas petromax was set in front of a white cloth measuring 1.5 m X 2 m. The cloth was fixed vertically between two poles or trees and the bulb was hanged in such a way that it reached nearly half the length of the cloth. Stretching, tagging, genitalia dissection, photography and identification of moths from family Geometridae were done. Environmental and habitat variables collected during sampling were entered and prepared for ecological analysis. Literature references related to climate change and Himalayan Lepidoptera from Zoological Records were collected from the year of 2008, 2011, 2014 and bibliography was prepared.

Selected for Ph.D. program at Saurashtra University, Gujarat and completed course work (from 02.01.2017 to 23.03.2017) at Wildlife Institute of India. Ph.D. synopsis on the topic of “Diversity and Distributional Pattern of Family Geometridae in GHNP Conservation Area” has been prepared and submitted to Saurashtra University.

Uttaran Bandyopadhyay, JRF

Two reconnaissance surveys were done in Askot Wildlife Sanctuary, Uttarakhand and in the months of August-September, 2016 and November-December, 2016. Moths were sampled from different sampling area based on elevation, vegetation type and land-use practices following Stratified Random sampling framework. For this, along a transect ranging from lowest to highest altitude; random sites were selected at every 300m vertical distance. Two or three sampling sites were selected at every habitat for encountering ample representation of different moth species. Small clean vials or glass jars were used as killing bottles in which Ethyl Acetate or Benzene were used as killing agent. Collected specimens were kept in insect envelopes made of oil paper with the proper label of date and site of collection.

Stretching, tagging, genitalia dissection, photography and identification of Moths from family Noctuidae were done. Literature references related to climate change and Himalayan Lepidoptera from Zoological Records from the year of 2010, 2012, 2013 were collected and bibliography was prepared.

180 Species of Family Notodontidae with detailed distribution information from secondary literature entered in the catalogue format for preparing Indian Notodontidae checklist.

Selected for Ph.D. program at Saurashtra University, Gujarat and completed course work (from 02.01.2017 to 23.03.2017) at Wildlife Institute of India. Synopsis writing for Ph.D. on the topic of “Diversity and Distribution Pattern of Moths (Lepidoptera: Heterocera) with special emphasis on Family Noctuidae in Askot Wildlife Sanctuary, Uttarakhand” has been prepared and submitted to Saurashtra University.

Rahul Ranjan, JRF

As no research permission in Kangchendzonga National Park, Sikkim was obtained from the concerned Forest Department, field sampling couldn’t be initiated here. The fellow was engaged in processing and identification of samples collected from different study sites. Leg samples were chopped and processed for further molecular studies. Identified specimens were coded with unique NMH ID. Moreover, specimens of family Pyralidae, Geometridae, Lasiocampidae and Erebidae were identified following standard taxonomic procedure.

Kamalika Bhattacharyya, JRF

Reconnaissance survey for selecting permanent sampling sites were undertaken in Central Himalaya through three field trips during the period of August-September and November-December, 2016 in Singalila National Park and Neora Valley National Park. The period covered four season – pre- monsoons, monsoon, post-monsoons and winter. A stratified random sampling was conducted along an altitudinal gradient by categorizing the entire gradient into distinctive altitude and vegetation envelops primarily based on altitude, vegetation types, and land-use practices. Within each envelop random sampling sites was selected based on micro habitat parameters. 20 trap sites were selected at each sampling sites in order to obtain comprehensive representation of moth species assemblage. The selected sites were positioned at the approximate centre of plots with a homogeneous vegetation cover, so that moth catches at weak light sources largely reflect the local communities. The minimum distance of 50m between nearest trap sites was maintained, taking care that the lights are not visible from nearest sites. Trap sites were chosen such that the immediate vicinity of the trap provide shelter from wind, and giving maximum possible light catchments area. Details of the selected sampling sites and specimens collected are given in Table 5, page 8. Overall, 3339 samples were collected from the study sites during these surveys. Of these 550 samples had been studied for taxonomic identification and 121 of these had been processed for DNA extraction for molecular taxonomic studies.

Selected for Ph.D. program at Saurashtra University, Gujarat and completed course work (from 02.01.2017 to 23.03.2017) at Wildlife Institute of India. Synopsis writing for Ph.D. on the topic of “Moth Assemblages along Altitudinal Gradient in SNP and NVNP- An Ecological and Molecular Approach Emphasizing on Noctuidae” has been prepared and submitted to Saurashtra University.

Subrata Gayen, JRF

Preliminary surveys for selecting sampling sites undertaken in Dihang-Dibang Biosphere Reserve during August-September, 2016 and November-January, 2016. Currently undergoing field survey around Anini, Mippi and Dumdim within Dibang National Park. So far 785 Lepidoptera specimens were collected to assess the species and taxonomic diversity of Dihang Dibang Biosphere reserve and targeted four different routes to assess environmental factors and species diversity. Those routes are having the highest altitudinal gradients up to 4000 m in Eastern Himalaya. All collected specimens have been stretched, tagged and photographed for further taxonomic study. The processed data has been gathered in Excel Workbook (BOLD format). Validation of Superfamily Pyraloidea recorded from Indian Himalayan region for generating annotated list of moths. Literature review done on genus Abraxas (Family Geometridae) for the taxonomy of collected specimens. From the present collection, a total of 95 specimens of 40 species under 35 genera in 10 families have so far been identified.

Gaurab Nandi Das, JRF

Initial surveys for butterfly inventory are being undertaken in following period: Singalila National Park during August-September, 2016, November to January, 2017 in Dihang Dibang Biosphere Reserve (DDBR), and ongoing survey in DDBR from March, 2017. Data on species richness and abundance of butterflies in respective altitudinal zones were collected on 500m transects which were walked for 30 minutes each under sunny and open sky, when the cloud cover was less than 70%. All butterflies seen during the transect walk in an imaginary 5x5x5 (m) box around the observer were recorded. Butterflies that were too fast or too distant to reliably identify during flight were not counted. Butterflies that could not be readily identified visually were either photographed or captured using hand-held sweep net and were released after identification. The very rare or difficult to identify species were captured and taken back to laboratory for further identification process. Thus, from Singalila NP, 18 species of butterflies were recorded with photographic evidence. Among them, mostly dominated genera Lethe with 6 species, of which Lethe atkinsonia, Lethe maitrya and Lethe goalpara were recorded. From DDBR, about 139 species of butterflies were recorded in 17 transects, among which most interesting findings are Drupadia scaeva cyara, which is first time record from Arunachal Pradesh, Eastern Himalaya. Additionally, 297 secondary literatures on butterflies of Indian Himalayan Region were screened and analysed which resulted in a total of 1187 subspecies/985 species under 337 genera belonging to 6 families (Table 8, Figure 11).

Sumantika Chatterjee, JRF

DNA isolation has been carried out for 500 samples using organic phase extraction method/silica column kit method/and magnetic bead based protocol. Out of 500 samples, 468 samples were successfully amplified for about 648 base pairs (bp) from the 5' end of the mtCOI gene using primer pair LCO-HCO. The amplified PCR products of 468 samples were checked in 1.2% agarose gel. The PCR amplified products were purified using QIAquick Gel Extraction Kit (Qiagen, Valencia, CA) following the manufacturer’s protocols. Approximately, 15 ng of the purified PCR product for each of 468 samples was used for sequencing. The cycle sequencing was performed with BigDye®Terminator ver. 3.1 Cycle Sequencing Kit (Applied Biosystems, Inc.) using 3.2 Picomoles of both forward and reverse PCR primers on a ABI thermal cycler with following parameters: 96 °C for 1 min, then followed by 25 cycles of 96 °C for 10 s, 50 °C for 5 s and a final extension at 60 °C for 1 min15 s. The cycle sequencing products were cleaned by using BigDye X-terminator kit (Applied Biosystems Inc.) and loaded on 48 capillary ABI 3730 Genetic analyser. The SeqScape software version 2.7 (Applied Biosystems Inc.) was used to analyze the forward and reverse chromatograms to obtain the consensus sequences.

The generated forward and reverse sequences were aligned with the representative barcode sequences from global database for similarity search. The developed sequences were submitted to BOLD database under the project ‘Lepidoptera of Indian Himalayas’ for acquiring the BOLD-IDs (details provided in Table).

Angshuman Raha, JRF

Initial collection survey was undertaken in GHNP in August-September, 2016. Primary identification of 1464 number of moth specimens were done through studying external morphology like wing venation, wing markings and spots, head and leg characters. Literatures consulted for identification were: Hampson (1892-1896), Holloway (1987-2014), Haruta (1997-2003), Moths of Thailand; and online resources like: www.mothsofindia.org, www.jpmoth.org, www.africanmoths.com. Further, genitalia dissection was done for confirmation of species whose external morphological characters were not sufficient for species-level identification. For genitalia study, methodology provided in Robinson (1976) was adopted. The detached abdomens were kept in 10% KOH overnight for dissolving away the muscles and unwanted parts. Soaked abdominal segments were dissected in 20% ethyl alcohol for taking out genitalia which were further studied and photographed under Leica S8AP0 HD binocular microscope. After proper dehydration, the genitalia were stored in vials containing a mixture of alcohol and glycerol in the ratio of 3:1. Nearly 100 specimens of family Noctuidae, Notodontidae and Geometridae were dissected for genitalia study (identified species list has been provided in Table 7).

Rohit Kumar Jaiswal, JRF

Initially Toposheets for all the study sites were acquired from Survey of India. Initial boundary was demarcated for all the PAs along with sampling routes and points marked at every 200 m according to vegetation type. Satellite data for Digital Elevation Model (SRTM 1 Arc-Second Global) have been downloaded in Georeferenced Tagged Image File Format (GeoTIFF). DEM, Contour map for 200m interval and Aspect map with the 2m grid size have been prepared for all PAs (Fig. 1,3,5,7 & 8). For NDVI, Landsat 8 and MODIS data downloaded from United States Geological Survey (USGS) and climatic variable data downloaded from WORLDCLIM for different temporal scale.

Preliminary surveys for ground-truthing were undertaken in Singalila National Park, Askot Wildlife Sanctuary and Great Himalayan National Park and Dihang-Dibang Biosphere Reserve.

The prediction of spatial species distribution of threatened Apollo butterflies in Indian Himalayan Biogeography zones are been carried out using MAXENT (Maximum Entropy), which uses species presence-only record data and climate variable data to predict the environmental suitability for the particular species. The model uses mathematical algorithm to analyse association between species and environmental conditions. The presence-only record data was collected from secondary literature and from Global Biodiversity Information Facility (GBIF). The data processing and analysis are being performed on Arc-GIS and “R” platform. Since evaluation or validation is critical and important for modelling study, so in this study cross validation procedure like k-fold partitioning or subsetting for model evaluation and validation have been adopted.

Table 4. Fellowship Description at Institutional/ University Level 1. No. of Research Publications: Published: 01 “Geometridae Stephens, 1829 from different altitudes in Western Himalayan Protected Areas of Uttarakhand, India” in SHILAP Revta. lepid., 44 (176) December 2016: 21: ISSN: 2340-4078, ISSN: 0300-5267. Under Review: 01 “Record of Drupadia scaeva cyara Hewitson, 1878 (Lycaenidae : Lycaeninae : Theclini) from Lower Dibang Valley, Arunachal Pradesh, India” in Journal of Threatened Taxa. Under Preparation: 07  “Preliminary records of moths from different altitudes in Askot Wildlife Sanctuary, Uttarakhand”  “On a collection of moths from Namdapha National Park, Arunachal Pradesh”  “Moths of Indian Himalayan Region: Faunal overview & status and gap in our current knowledge”  “Taxonomic studies on the Phlogophora sp. (Noctuidae) collected from Indian Himalayan region with two new records from India”  “Preliminary records of moths from different altitudes in Central Himalayan landscape of West Bengal”  “Catalogue of Indian Notodontidae with three new records from India”  “An annotated list of moths from Indian Himalayan Region”

2. No. of Data Sets generated: 03  Project Console-Lepidoptera of Indian Himalayas [ZSILH]: Sequence data of 368 specimens, of 39 identified species, containing Geometridae (199 sequences), Noctuidae (56 sequences), Erebidae (45 sequences), Crambiade (34 sequences).  Validated & geo-referenced Lepidoptera species records from Indian Himalayan Region  Geo-referenced Apollo species records from Indian Himalayan Region

3. No. of Conferences/ Workshops attended: 01 On “Workshop on Species Distribution Modelling in MAXENT”, organized by Indian Statistical Institute, Kolkata from 09th to 14th January, 2017, by Rohit Kumar Jaiswal. Species distribution modelling was demonstrated in R platform. MAXENT species distribution modelling was taught in Arc GIS & DIVA GIS.

4. No. of Sites/ Study Area Covered: i. Hemis National Park, Jammu & Kashmir (1 gradient explored) ii. Great Himalayan National Park, Himachal Pradesh (4 gradients explored) iii. Askot Wildlife Sanctuary, Uttarakhand (2 gradients explored) iv. Singalila National Park, West Bengal (1 gradient explored) v. Neora Valley National Park, West Bengal (2 gradients explored) vi. Dihang-Dibang Biosphere Reserve, Arunachal Pradesh (3 gradients explored)

5. No. of Best Practices suitable for IHR: NA

6. New Observations/ Innovations: Family Notodontidae: Three species, each from genera Phalera Hübner, 1819 (Phalerinae), Clostera Samouelle, 1819 (Pygaerinae) and Hiradonta Matsumura, 1924 (Ptilodoninae) are being recorded as new records to India. Moreover, three species, each from genera Allodonta Staudinger, 1887 (Ptilodoninae), Honveda Kiriakoff, 1962 (Biretinae) and Ginshachia Matsumura, 1929 (Pygaerinae) have been found interesting with the probability of becoming new to science. Family Noctuidae: Two species of Phlogophora, P. meticulodina (Prout, 1950), P. subpurpurea Leech, 1900 has been recorded for the first time from India. Family Geometridae: Two species, each from genera Chiasmia Hübner, 1823 (Ennominae) and Hyposidra Guenée, 1857 (Ennominae) have been found new to India.

LEPIDOPTERA (INSECTA) AS POTENTIAL INDICATOR-TAXA FOR

TRACKING CLIMATE ANNUAL REPORT CHANGE IN THE INDIAN HIMALAYAN 2016-17 LANDSCAPE

(1st April 2016-31st March 2017) Zoological Survey of India, NMHS Fellowship Program Kolkata

Contents

Sl. No. Topic Page No.

I. Background of the Project 1 II. Objective-wise Progress 1  Objective I: Field sampling 1  Objective I: Species Identification & Morpho-Taxonomy 12  Objective II: Compilation of Secondary Database 25  Objective III: Secondary Data Compilation of Apollos 29  Objective III: Apollo Species From Primary Field Work 29  Objective IV: Methodology Adopted for DNA Barcoding 30 III. DETAIL OF SEQUENCE SUBMITTED TO BOLD 38 List of Plates Plate 1: Genitalia study of Family Notodontidae 14 Plate 2: Genitalia study of genus Phlogophora (Family Noctuidae) 15 Plate 3 & 4: Different Habitat Types Sampled 33-34 Plate 5-7: Moths Recorded from Different Himalayan Landscape 35-37

Himalayan Lepidoptera Project

I. Background of the Project Lepidoptera are considered as ‘flagships’ in insect conservation. Besides their popularity in human perception for nature appreciation, their biotic and abiotic specifications make them a suitable taxon for ecological entomology and habitat conservation studies. Being the third largest insect order with 1,57,424 species known worldwide, they are mostly phytophagous and thus show stochasticity according to the floral perturbations. Several aspects of climate change, such as elevated CO2 and O3 concentrations, elevated temperature and UV radiation, and changes in rainfall patterns directly affect host plant abundance and quality, which in turn disturbs physiology, behaviour and life history strategies and ultimately reduction in abundance, richness and diversity of Lepidopteran fauna. Indian Himalayan Region (IHR) due its habitat mosaics ranging from dry and rugged terrain of trans and western side to the lush cloud forests of eastern Himalaya harbours rich Lepidoptera fauna including many rare and endemic species. Vulnerability of IHR to continuous exasperation of climate change makes it of immediate concern that a thorough assessment of diversity and distribution pattern of Himalayan Lepidoptera is need of the hour, which will assist in laying the foundation for future monitoring programme in this fragile landscape. II. Objective-wise Progress Objective I: To investigate the differential processes influencing the diversity and distribution pattern of Lepidoptera assemblages (moths and butterflies), thus analysing influence of biogeography, suit of biotic-abiotic factors and anthropogenic disturbance on Lepidoptera diversity along altitudinal gradient in six Himalayan Biogeographic Provinces . Field Sampling Altogether following field sampling sessions were undertaken: Table 1: Brief detail of surveys conducted till date. Biogeographic Province Site Survey Period Specimens Collected Trans Himalaya (1A) Hemis National Park, Jammu & 07.ix.2016 - 17.ix.2016 49 Kashmir North-Western Great Himalayan National 25.viii.2016 - 24. viii.2016 891 Himalaya (2A) Park, Himachal Pradesh 04.xii.2016 - 15.xii.2016 Western Himalaya (2B) Askot Wildlife Sanctuary, 18.ix.2016 – 28.ix.2016 486 Uttarakhand 24.xi.2016 - 02.xii.2016 Central Himalaya (2C) Singalila National Park, West 20.viii.2016 - 15.ix.2016 3339 Bengal 06.xi.2016 – 10.xi.2016 29.xi.2016 - 11.xii.2016 Eastern Himalaya (2D) Dihang Dibang Biosphere 20.viii.2016 -15.ix.2016 501 Reserve, Arunachal Pradesh 29.xi.2016 - 10. i.2017 22.iii.2016 - Ongoing *Fieldwork in Tibetan Plateau (1B) in Kangchendzongha Biosphere Reserve, Sikkim can’t be initiated due to permission issue

Himalayan Lepidoptera Project

Trans-Himalaya, Hemis National Park (HNP), Jammu & Kashmir Research Objectives • To investigate the differential processes influencing the distribution pattern of Lepidoptera assemblages (moths and butterflies) of Hemis National Park • Generation of robust datasets generated through ecological monitoring at habitat level The reconnaissance survey was conducted in the vicinities of Hemis Village, Sumdo, Nagdang Village, Ngdang Pho-I, Nagdand Pho-II and Nagdang Pho-II within the Hemis National Park. After the reconnaissance survey Seven sampling site has been chosen as permanent sampling sites - Sumdo, Nemaling, Nagdang village, Nagdang Pho-I-II-III and Markha, from inside the boundary of national park. At every sampling site, at least 2-3 sampling points has been chosen for repetitive sampling based on habitat type, elevation, and habitat quality and species abundance data from the survey. Specimen collection details have been provided in Table 2. Light trapping for moths and Random Survey for butterflies were conducted at every site to get the abundance data. Specimens were collected even in the absence of collection permission at the time of survey. Approximately 21 moth specimens were collected from Hemis Village, outside the national park boundaries and 20 specimens from inside the national park. Total 41 moth specimen have been collected from the National Park. Many Butterflies species including Apollo has been sighted in the area. The village Sumdo, which means the base point or gateway of 3 different valleys was the main basepoint of this current survey. The research team talked with local villagers about different issues pertaining to Lepidoptera diversity and agriculture practise. According to local people, plenty of butterfly species has generally been sighted during the month of June to August. The mountain of the village has good pasture land which are well-watered and thus different species of flowering plants grow here which makes it a suitable habitat for butterflies, even in extreme winter months. According to villagers there are some illegal trading of butterfly specimens by some foreigner who usually visit the area for recreational and trekking purpose. People expressed their concern on receding ice caps from the top of the mountains and resultant decline in water availability which is one of the main cause of declining butterfly numbers in recent years. Although the team got very limited time for field activity, a good proportion of Lepidopteran species pool was recorded during the course of the study. It was noted that the major Lepidopteran species were active around village agricultural patches and these habitats are stronghold for local species diversity in the resource-poor seasons. Local people take considerable pride for the wildlife species inhabiting the area and there is a nice scope to involve local people in conservation management practices, using the Apollo butterflies as flagships for Lepidoptera conservation. The team proposes to use questionnaire survey to assess socio-economic backdrop of Apollo conservation focussing on their role as pollinators for major crops, illegal trading of the species and perception of local communities for formulating effective conservation management strategies. Creating awareness among local communities and forest managers on the importance of Lepidoptera as a tool for monitoring climate change and habitat alterations is an urgent task in this climatically fragile Trans- Himalayan landscape.

Himalayan Lepidoptera Project

Table 2: Specimen collection and details of survey area. No. of No. of Elevation Moth Butterfly Sites Lat. Long. Remark (m) Samples Samples Collected Collected Hemis N33⁰54’ E77⁰44’ Sample collection; Light 3601 21 Village 39.6” 39.9” 2 Trapping N33⁰51’ E77⁰42’ 3 Sample collection; Light Sumdo 3690 4 15.0” 28.1” Trapping; Area Within HNP Nagdang N33⁰52’ E77⁰39’ 2 Sample collection; Light 3942 16 Village 00.9” 34.6” Trapping; Area Within HNP 0 Apollo Sighted, Data taken on Nagdang N33⁰52’ E77⁰38’ 4126 0 vegetation & topography. Pho-I 20.8” 18.6” Suitable Habitat for Butterflies. Nagdang N33⁰52’ E77⁰37’ 1 4237 0 Pollard Walk Transect Pho-I I 37.7” 32.1” 0 Random sighting; Well-watered Nagdang N33⁰52’ E77⁰36’ 4348 0 pasture land for livestock Pho-III 44.6” 51.9” grazing

Figure 1: Sampling Sites in Hemis National Park; DEM and Contour Map & Aspect Map of HNP

Himalayan Lepidoptera Project

North West Himalaya, Great Himalayan National Park (GHNP), Himachal Pradesh Research Objectives • To investigate the differential processes influencing the distribution pattern of Lepidoptera assemblages (moths and butterflies) of Great Himalayan National Park • Generation of robust datasets generated through ecological monitoring at habitat level Two reconnaissance surveys were done in August and December in which a total of four trek routes (sampling gradients) have been selected. About 650 specimens of Moth have been collected from GHNP and the adjacent areas. Live photograph of all moth specimens were taken in the field. All specimens were sorted and stretched. From the August tour, 11 species of butterflies have been recorded with photographic evidence. Among them, two very rare species of Apollo butterflies have been found. Table 3: Details of the surveys conducted at GHNP. Number of Sampling Longitude Altitude (m) Moth Latitude Sites Date samples Collected Ropa 26.08.16 31.76558 N 077.35765 E 1515 313 Lapah 27.08.16 31.77089 N 077.42718 E 2247 47 Dhel 29.08.16 31.75681 N 077.46146 E 3567 226 Shakti 30.08.16 31.78847 N 077.49486 E 2258 75 Thati 08.09.16 31.73912 N 077.35873 E 2917 10 Padhar 06.09.16 31.74993 N 077.37323 E 2437 26 Thinhi 07.09.16 31.72932 N 077.36654 E 3226 6 Maror 15.09.16 31.78827 N 077.53188 E 2480 59 Parkachi 17.09.16 31.79900 N 077.59122 E 2975 6 Ropa 04.09.16 31.76558 N 077.35765 E 1515 0 Gajnau 08.09.16 31.74055 N 077.37722 E 2607 87 Thihni 09.09.16 31.72930 N 077.36652 E 3224 23 Bhagisari 10.09.16 31.81105 N 077.38952 E 2435 13

400 356 350 300 250 200 150 102 108 100 72

Number of specimens of Number 50 2 3 1 3 3 9 2 0

Figure 2: Family-wise abundance pattern of moths from GHNP

Himalayan Lepidoptera Project

Figure 3: Sampling Sites in GHNP; DEM and Contour Map & Aspect Map of GHNP

Himalayan Lepidoptera Project

Western Himalaya, Askot Wildlife Sanctuary (AWLS), Uttarakhand Research Objectives • To investigate the differential processes influencing the distribution pattern of Lepidoptera assemblages (moths and butterflies) of Askot Wildlife Sanctuary • Generation of robust datasets generated through ecological monitoring at habitat level Two reconnaissance surveys were undertaken in September and November in which two trek routes (sampling gradients) have been selected. About 570 specimens of Moth have been collected from AWLS and the adjacent areas. Live photograph of all moth specimen was taken in the field. All specimens were sorted and stretched. From the August-September tour, 12 species of butterflies have been recorded with photographic evidence. One rare and important species of Apollo butterfly was also recorded from the region. Table 4: Details of the surveys conducted at AWLS. Number of Sampling Altitude (m) Latitude Longitude Moth samples Sites Date Collected Gowalghat 22.09.16 29.91536 N 080.4032 E 2314 72 Vayman 23.09.16 29.93124 N 080.38678 E 3218 71 Vatyakhan 24.09.16 29.94033 N 080.39720 E 3390 38 Ringal Fst 25.09.16 29.92458 N 080.39169 E 2932 66 Baram Pol 26.09.16 29.86902 N 080.37186 E 1121 285 Jauljivi 19.09.16 29.75087 N 080.37885 E 632 37 Jauljivi 27.11.16 29.75087 N 080.37885 E 764 11 Ganagaon 24.11.16 29.75440 N 080.38208 E 2480 59 Naryan Ar 29.11.16 29.97136 N 080.65444 E 2536 0 Payang 30.11.16 29.98000 N 080.65938 E 2657 0 Lankari 1.12.16 29.97677 N 080.67483 E 2250 0

180 168 160 140 123 120 100 79 80 60 43 specimenof No. 40 20 1 1 3 5 5 1 2 0

Figure 4: Family-wise abundance pattern of moths from AWLS

Himalayan Lepidoptera Project

Figure 5: Sampling Sites in AWLS; DEM and Contour Map & 3-D Map of AWLS

Himalayan Lepidoptera Project

Central Himalaya, Singalila & Neora Valley National Park (SNP/NVNP), West Bengal Research Objectives • To investigate the differential processes influencing the Lepidoptera diversity (moths and butterflies) of Singalila National Park • Generation of robust datasets generated through ecological monitoring at habitat level Reconnaissance survey of the study site for selecting permanent sampling sites were undertaken through three field trips during the period of August-September and November- December, 2016. The period covered three season – monsoon, post-monsoons and winter. 137 samples of lepidopterans were collected in the first survey from outside the national park boundary. 121 moths and 6 butterflies were collected during repeat sampling conducted in November- December. Samples have been sorted, stretched and processed for further taxonomic studies. 1222 samples of lepidopterans were collected during reconnaissance survey in upper and lower NVNP in August-September, 2016. Table 5: Details of the surveys conducted at SNP/NVNP. Study area Sampling Sites Co-ordinate Altitude Habitat Date Samples (m) Collected Singalila NP Gairibas N27.0427°/E88.0371° 2600 Temperate 2.12.16 05 Tumbling N 27.03147°/ E88.06885° 2914 Temperate 28.8.16 NA Tonglu N 27.034147°/E 88.079398° 3049 Temperate 29.8.16 Kalpokhari N27.0783°/E88.0187° 3021 Temperate 3.12.16 05 Sandakphu N27.1061°/E88.0024° 3540 Alpine 1.9.16 Lamahatta N27.0495°/E 88.3427° 1795 Subtropical 5.12.16 33 Chitra N26.9913°/E88.1119° 2366 Subtemperate 3.9.16 137 Srikhola N 27.1304°/E 88.0735° 1931 Subtropical 8.12.16 110 Neora Valley Suntali Khola N27.01042 E88.78983 751 Subtropical 9.11.16 3184 NP Mouchaki camp N27.0268°/E88.7864° 1186 Subtropical 7.11.16 102 Rishop N27.1068°/E88.6495° 2591 Subtemperate 5.9.16 675 Lava N27.0863°/E88.6615° 2200 Subtropical 7.9.16 547

180 160 140 120 100 80 60

40 20 No. of Individuals recorded Individualsof No. 0

Figure 6: Family-wise abundance pattern of moths from Central Himalaya

Himalayan Lepidoptera Project

Figure 7: Sampling Sites in SNP; DEM and Contour Map of Singalila National Park

Himalayan Lepidoptera Project

Eastern Himalaya, Dihang Dibang Biosphere Reserve (DDBR), Arunachal Pradesh Research Objectives • To investigate the differential processes influencing the distribution pattern of Lepidoptera assemblages (moths and butterflies) of Dihang Dibang Biosphere Reserve • Generation of robust datasets generated through ecological monitoring at habitat level Primary survey was conducted, between 20th August to 20th September 2016, with gradient transect walk and quadrat sampling started from 449 m to 2163 m altitude for reconnaissance of Lepidopteran diversity. Light-trap stations were installed at different altitude at Roing, Mayodiya, Anili, Theei, Cheepe for documentation of moth diversity. Extensive quadrat sampling was not possible due to heavy rainfall. A total of 341 individuals of moth species were documented through photography and 216 individual specimens were collected. Other parameters like weather parameter, humidity, temperature, canopy and cloud cover data had been recorded. In continuation to the previous field survey conducted during, we resampled previously marked points and established new light-trap stations between 29th November 2016 to 10th January 2017. We obtained 285 moth specimens and the species identification is in process for collected specimens. As the survey was conducted in peak winter, nocturnal Lepidoptera activity was not much, with minimum temperature of 5.2⁰ C at which we got moth samples. During this survey, we recorded and documented 139 butterfly species. Nymphalidae was the most dominant butterfly family followed by Lycaenidae, Pieridae, Hesperiidae and Papilionidae, in this survey. Drupadia scaeva cyara is the first-time record from Arunachal Pradesh, Eastern Himalaya, and recorded after a gap of 83 years from India (last recorded by Eliot, 1934 from Gangtok). This is also first photographic evidence from mainland India. Table 6: Details of the surveys conducted at DDBR. SL. Site Date of Longitude Latitude Altitude Temp Humidity No. of No survey (m) (℃) (%) Specimen . collected 1 Roing 23.08.16 28.09.24.9 95.5.50.03 607 37 99 99 2 Myodia 25.08.16 28.13.59.16 95.54.29.3 2463 35 82 82 3 Engalin 29.08.16 28.33.23.91 95.37.16.6 1627 34.2 64 64 4 Cheppe 31.08.16 28.36.38.2 95.29.47.1 1987 35.2 98 98 5 Point 73 06.12.16 28.30.30.2 95.49.04.0 826 13.33 56 17 6 Anelih 08.12.16 28.31.41 95.46.18.5 868 15.3 60 48 7 Ch_800 09.12.16 28.32.09.3 95.44.24.9 830 15.9 64 26 8 Ch_600 10.12.16 28.32.19.2 95.44.00.2 664 11.8 62 24 9 Ch_820 12.12.16 28.32.29.5 95.39.54.6 820 13.2 64 11 10 Ch_900 13.12.16 28.32.66.00 95.38.724 836 9.1 62 7 11 Chepe 16.12.16 28.36.560 95.29.874 1552 9.93 65.66 15 12 Ch_1900 18.12.16 28.37.678 95.29.158 1900 8.33 63.66 15 13 Ch_2100 19.12.16 28.37.849 95.29.318 2110 9.4 63.66 42 14 Ch_1200 21.12.16 28.34.234 95.32.655 1278 9.2 62 43 15 Ch_925 22.12.16 28.32.601 95.37.871 925 14.16 68 14 16 Idulli 07.01.16 27.59.525 95.49.623 147 14.9 77 23

Himalayan Lepidoptera Project

Figure 7: Sampling points and DEM of Dihang-Dibang Biosphere Reserve

Himalayan Lepidoptera Project

. Species Identification & Morpho-Taxonomy  So far, across the entire Himalayan landscape, 4863 number of specimens have been collected, stretched and labelled for further identification process.  Among them, 1464 number of specimens have been identified which comprises 507 morpho- species under 265 genera (The identified genera list with number of species and specimens identified are provided in Table 7).  Primary identification was done through studying external morphology like wing venation, wing markings and spots, head and leg characters. Literatures consulted for identification were: Hampson (1892-1896), Holloway (1987-2014), Haruta (1997-2003), Moths of Thailand; and online resources like: www.mothsofindia.org, www.jpmoth.org, www.africanmoths.com.  Further, genitalia dissection was done for confirmation of species whose external morphological characters were not sufficient for species-level identification. For genitalia study, methodology provided in Robinson (1976) was adopted. The detached abdomens were kept in 10% KOH overnight for dissolving away the muscles and unwanted parts. Soaked abdominal segments were dissected in 20% ethyl alcohol for taking out genitalia which were further studied and photographed under Leica S8AP0 HD binocular microscope. After proper dehydration, the genitalia were stored in vials containing a mixture of alcohol and glycerol in the ratio of 3:1.

800 Genera Species Specimen 697 700

600 547 500

400

300 245 220 200 158 158 107 88 104 100

0 W & NW Himalaya Central Himalaya Eastern Himalaya

Figure 9: Site-wise collection and identification details

Himalayan Lepidoptera Project

80 Western Central Eastern 70 75

30 32 31 20 22 19 10 13 1 3 4 2 11 8 5 9 7 0 Hesperiidae Papilionidae Pieridae Lycaenidae Nymphalidae

140 No.of Genera No. of Species

120

100

Nolidae Tinedae Cossidae Erebidae Pyralidae Eutelidae Uraniidae Noctuidae Crambidae Sphingidae Zygaenidae Saturniidae Callidulidae Drepanidae Thyriddidae Lasciocampi… Bombycidae Limacodidae Eupterotidae Geometridae Notodontidae

Figure 10: Family-wise identified species of butterfly and moth

 Nearly 100 specimens of family Noctuidae, Notodontidae and Geometridae were dissected for genitalia study. Five species of genus Phlogophora Treitschke, 1825 (Noctuidae) have been identified, out of which two species are being recorded for the first time from India. More than 20 specimens of 8 species under 7 genera of family Geometridae were dissected for identification confirmation. Two species, each from genera Chiasmia Hübner, 1823 (Ennominae) and Hyposidra Guenée, 1857 (Ennominae) have been found interesting which require further investigation. In Notodontidae, the genitalia dissection indicates 19 different species under 12 genera. Three species, each from genera Phalera Hübner, 1819 (Phalerinae), Clostera Samouelle, 1819 (Pygaerinae) and Hiradonta Matsumura, 1924 (Ptilodoninae) are being recorded as new records to India. Moreover, three species, each from genera Allodonta Staudinger, 1887 (Ptilodoninae), Honveda Kiriakoff, 1962 (Biretinae) and Ginshachia Matsumura, 1929 (Pygaerinae) have been found interesting with the probability of becoming new to science.

Himalayan Lepidoptera Project

Plate 1: Genitalia study of Family Notodontidae

Himalayan Lepidoptera Project

Plate 2: Genitalia study of genus Phlogophora (Family Noctuidae)

Himalayan Lepidoptera Project

Table 7: Number of species and specimens under identified genera of moths from Western & North-Western Himalaya, Central Himalaya and Eastern Himalaya

Western & North-Western Himalaya Central Himalaya Eastern Himalaya Family Genus No. of Species No. of Specimens No. of Species No. of Specimens No. of Species No. of Specimens Tinedae Zarba 1 1 Cossidae Cossus 2 4 Xyleuetes 1 1 Zeuzera 2 4 1 2 Limacodidae Altha 1 3 Chalcoscelides 1 8 Phocoderma 1 1 1 2 Zygaenidae Chalcosia 1 1 Gynautocera 1 1 1 2 Neochalcosia 1 1 Pidorus 2 4 1 1 Thyriddidae Banisia 1 1 Rhodoneura 1 1 Callidulidae Pterodacta 1 1 Pyralidae Endotricha 2 8 Orthaga 1 1 Orybina 1 1 2 2 Sacada 1 2 1 2 1 7 Crambidae Agathodes 1 1 1 8 Agrioglypta 1 4 Agrotera 1 3 Blepharomastix 1 1 Botyodes 1 8 3 5 Bradina 1 1 Cirrhochrista 2 2 Conogethes 1 1

Himalayan Lepidoptera Project

Cotachena 2 2 1 2 Cydalima 1 2 Cydalima 1 2 Endocrossis 1 1 Eoophyla 1 1 Filodes 1 1 Glyphodes 3 6 3 7 2 7 Heortia 1 2 Heterocnephes 1 2 Lamprophaia 1 1 Maruca 2 3 1 2 1 1 Nausinoe 1 1 1 2 Neadeloides 1 4 Pagyda 1 1 2 9 Palpita 1 11 2 14 Parotis 3 6 1 1 Pleuroptya 3 15 1 1 Pygospila 2 18 1 1 Rhimphalea 1 1 Spoladea 2 3 2 16 Syllepte 1 1 Synclera 1 2 Syngamia 1 5 Uncobotyodes 1 1 Drepanidae Cyclidia 1 1 Ditrigona 1 1 Gaurena 1 2 1 4 1 2 Habrosyne 1 1 1 1 Thyatira 1 1 Tridrepana 1 1 Lasciocampidae Euthrix 1 1 1 3

Himalayan Lepidoptera Project

Ganisa 1 1 Gastropacha 1 1 1 8 Kunugia 1 2 Metanastria 1 1 Paralebeda 2 4 Eupterotidae Apona 1 1 Eupterote 4 5 1 3 Bombycidae Mustilia 1 3 Triuncina 2 3 Saturniidae Acarontia 1 2 Actias 1 1 Antherea 1 1 Archaeoattacus 1 1 Leopa 1 1 1 5 Sphingidae Acherontia 1 1 Acosmeryx 1 1 1 1 Agrius 1 1 1 1 Amplypterus 1 1 Anambulyx 1 1 Clanis 2 2 Deilephila 1 1 Elibia 1 1 Euphanacra 1 1 Hippotion 2 2 1 5 Marumba 1 2 Polyptychus 1 1 Psilogramma 1 1 Rhagastis 1 7 Rhopalopsyche 1 2 Rhyncholaba 1 2 Theretra 1 14 5 11 1 4

Himalayan Lepidoptera Project

Uraniidae Epiplema 1 5 Geometridae Abraxas 8 21 5 136 4 16 Agathia 1 1 Alcis 1 2 Anonychia 2 19 Antrigodes 1 1 Aporandria 1 1 Arichana 4 65 1 1 2 4 Biston 1 1 1 1 Cidariplura 1 3 Chiasmia 1 2 1 2 Chlorissa 1 1 Chlorodontopera 1 2 Chlororithra 1 1 Chorodna 2 3 Chrysocraspeda 1 3 Comostola 1 2 Corymica 1 1 Ctenognophos 1 3 Dysphania 1 1 Ecliptopera 1 1 Electrophaes 1 1 Elophos 1 1 Erebomorpha 1 1 Eucyclodes 1 1 1 1 Eustroma 1 11 Fascelina 1 1 1 1 Heterolocha 2 2 1 1 Heterostegane 1 1 Hypochrosis 1 1 Krananda 3 4 1 2

Himalayan Lepidoptera Project

Lophpphelma 1 1 Loxaspilates 3 19 Luxiaria 1 1 Medasina 2 5 Milionia 1 1 Mixochlora 1 2 Odontopera 2 7 Ophthalmitis 1 1 Opisthograptis 2 2 Organopoda 1 1 Ornithospila 1 1 Ourapteryx 3 21 2 14 1 3 Parasynegia 1 1 Percnia 1 1 1 6 2 6 Photoscotosia 3 49 Phthonandria 1 1 Plutodes 1 1 1 2 1 1 Pingasa 3 4 Problepsis 2 5 1 1 Pseudomiza 1 1 1 1 Psilocambogia 1 1 Psyra 2 5 1 1 Rhodometra 1 1 Sarcinodes 1 1 Scopula 7 18 1 15 Tanaoctenia 1 1 1 4 Tanaorhinus 1 1 Thalassodes 1 1 Thinopteryx 1 1 Tristeirometa 1 1 Vindusara 1 1

Himalayan Lepidoptera Project

Xandrames 1 1 1 1 Zeheba 1 5 Notodontidae Acmeshachia 2 5 Clostera 1 4 1 1 Formofentonia 1 1 Ginschachia 1 1 Phalera 3 6 Ptilodon 1 1 Saliocleta 1 1 Stauropus 1 1 Syntypistis 1 1 Tarsolepis 1 1 Erebidae Achaea 1 1 Aegocera 1 1 1 1 Anisoneura 1 2 Arcte 2 3 Arctornis 1 4 Argina 1 2 Artena 1 2 Asota 3 5 1 2 Asura 1 1 Baorisa 1 1 Barsine 2 9 3 4 1 3 Bastilla 1 1 Bocula 1 1 Calesia 2 5 Carsina 1 1 Catocala 1 4 Chrysaegia 1 9 Chrysorabdia 3 7 1 1 Cispia 1 3

Himalayan Lepidoptera Project

Coleta 1 1 Conilepia 1 1 Cosmophila 1 1 Creatonotos 2 7 1 10 Cyana 8 24 4 16 1 3 Dasychira 1 1 Erebus 2 5 1 1 1 1 Ericeia 1 1 Eudocima 1 4 3 6 3 3 Euproctis 5 11 Fodina 1 1 Gandhara 2 5 Gazalina 1 1 Hamodes 1 2 Imaus 1 3 Ischyja 2 6 Lacera 1 3 Lemyra 1 1 Lyclene 5 17 3 8 1 1 Lygephila 1 4 Lymantria 6 12 2 5 1 1 Mecodina 1 1 Miltochrista 1 1 Nannoarctia 1 1 Numenes 1 3 1 2 Nyctemera 1 2 3 4 2 3 Oenistis 1 9 Olepa 1 1 Ophiusa 1 2 Pindara 2 3 Psilochira 1 1

Himalayan Lepidoptera Project

Pterogonia 1 1 Rema 1 1 Sarbanissa 1 1 Serrodes 1 2 Simplicia 1 2 3 3 Spilosoma 5 9 2 6 1 1 Spirama 2 6 Stigmatophora 1 2 Thyas 2 4 Tiruvaca 1 1 Utethesia 1 13 Vamuna 2 3 1 6 2 4 Xanthodes 3 7 Eutelidae Eutelia 1 2 Penicillaria 1 1 Nolidae Gabala 1 1 Gadirtha 1 4 Pseudoips 1 1 Risoba 1 3 2 4 Tyana 1 2 Westermannia 2 9 Noctuidae Achaeops 1 1 Acontia 1 1 Acronicta 1 2 Agrapha 1 2 Agrotis 1 1 Amphipyra 1 4 Anaplectoides 1 3 Apsara 1 1 Autographa 1 4 Blasticorhinus 1 1 1 1

Himalayan Lepidoptera Project

Western & North-Western Himalaya Central Himalaya Eastern Himalaya Family Genus No. of Species No. of Specimens No. of Species No. of Specimens No. of Species No. of Specimens

Callyna 1 1 Calopistria 1 3 2 3 Calyptra 1 1 Chasmina 1 2 Chrysodeixis 3 34 1 5 Conservula 1 1 Dichromia 3 8 Dictyestra 1 1 Diptherocome 1 8 2 10 Disepholcia 1 1 Haderonia 1 9 Mythimna 1 8 Nacna 1 1 Ochropleura 2 5 Oraesia 1 3 2 7 Ozarba 1 1 Paraxestia 1 1 Phlogophora 3 5 1 2 Polia 1 3 Polypogon 1 1 Polytela 1 1 Spodoptera 2 2 1 1 Total 245 697 158 547 104 220

Himalayan Lepidoptera Project

Objective II: Repeating monitoring surveys in historical collection localities originally carried out by earlier workers, and generating current species-distribution map from primary as well as secondary data, to see if there are any changes in species distribution range and generating predictive species distribution modelling for future climatic scenario.

. Compilation of Secondary Database: Initially literatures regarding diversity and distributional pattern of Lepidoptera done on GIS domain in the mountainous ecosystems worldwide have been searched and reviewed for better representation of data. To prepare the primary dataset, keyword searches for ‘climate change’, ‘Lepidoptera’, ‘Himalayan Biogeographic zone and provinces’, ‘Indian Himalayan states’ and ‘Lepidoptera (both moth & butterflies) inventory and distribution’ were made. Thus, a list of literatures was found most of which comprises State Fauna Series and Conservation Area Series published by Zoological Survey of India. Other than these, ‘Fauna of British India’ series are among the oldest literatures. Material examined or specimen details of species where ever available in these literatures were extracted and entered in MS Excel to prepare primary dataset. In excel sheet columns named Family, Subfamily, Genus, Species, Author, Locality or site, State, Latitude, Longitude, Altitude, Examples, Collector and Source were entered. From the literature review part of every specimen the details of these properties were put in the respective columns. For different dates and different localities in particular state separate entries were made for the same species. Since most of the studies were done earlier when devices like GPS were not used, coordinates for the collection localities were not being provided in these literatures. In some cases, only the altitudes of the collection sites were provided. These sites were searched in the internet for geographic coordinates. In case of sites whose coordinates were not available in the internet, or the sites have different names in past and present and thus couldn’t be accurately located, coordinates of the nearby towns, cities, localities etc. were considered and included in the dataset. From this dataset, a species list including state-wise distribution and altitude records of each species of the IHR was prepared for further analyses.

Butterfly: Total 297 secondary literatures were screened and analysed. After extensive literature review, a total of 1187 subspecies/985 species under 337 genera belonging to 6 families have been reported.

800 700

600 500 400

300 200 100

0 Jammu & Himachal Uttarakhan West Arunachal Sikkim Kashmir Pradesh d Bengal Pradesh Genus 99 142 203 262 216 266 Species 201 283 448 686 491 692 Subspecies 245 291 455 709 494 725 Figure 11: State-wise distribution of Butterfly genera, species and subspecies in IHR

Himalayan Lepidoptera Project

Table 8: Number of genera, species and subspecies of butterflies under different families and subfamilies.

Family Subfamily Genus Species Sub-Species Hesperiidae Coeliadinae 5 19 Pyrginae 22 64

Hesperiinae 53 125 80 208 221

Papilionidae Papilioninae 7 58 Parnassiinae 2 17

9 75 120

Pieridae Pierinae 16 48 Coliadinae 6 25 22 73 88

Riodinidae Riodininae 4 12 18 Lycaenidae Poritiinae 2 3

Miletinae 6 8

Curetinae 1 2 Lycaeninae 98 253 107 266 304 Nymphalidae Danainae 5 18

Satyrinae 33 139

Calinaginae 1 4 Amathusiinae 7 12 Nymphalinae 63 156

Charaxinae 3 16 Acraeinae 2 4

Libytheinae 1 2 115 351 436 Total 337 985 1187

Of 985 butterflies of Himalaya, 274 (28%) species and subspecies are listed under Indian Wildlife (Protection) Act, 1972 (WPA) of which 73 species come under schedule I, 191 species come under schedule II and 10 under schedule IV. In WPA, maximum species and subspecies names are taxonomically outdated, but here we provide numbers according to current valid names of species and subspecies. As per Gupta & Mandal (2005), 74 species also come under Red Data Book of Indian Butterflies. A substantial portion of Himalayan butterflies are localised either in one or two Himalayan states may be due to some ecological barriers or phylogenetic history, thus clearly indicating the conservation priority of these species and their habitats in this fragile landscape.

Himalayan Lepidoptera Project

Moth: After screening 93 secondary literature, 3533 validated species, 4 times higher than currently known butterfly species, are known from Indian Himalayan Region, which is distributed over 1234 Genera across 39 Families.

Table 9: Genera and species number of different families of moth recorded from IHR.

Sl No. Family No. of Genus No. of Species 1 HEPIALIDAE 4 8 2 PSYCHIDAE 1 2 3 TINEIDAE 6 15 4 GRACILLARIIDAE 2 2 5 YPONOMEUTIDAE 1 1 6 PLUTELLIDAE 2 4 7 LYONETIIDAE 1 1 8 AUTOSTICHIDAE 2 2 9 LECITHOCERIDAE 5 6 10 OECOPHORIDAE 6 7 11 COSMOPTERIGIDAE 1 1 12 GELECHIIDAE 2 3 13 PTEROPHORIDAE 9 17 14 IMMIDAE 1 2 15 TOTRICIDAE 21 35 16 COSSIDAE 6 26 17 SESIIDAE 5 14 18 LIMACODIDAE 19 79 19 ZYGAENIDAE 32 102 20 THYRIDIDAE 10 33 21 HYBLAEIDAE 1 1 22 CALLIDULIDAE 6 6 23 PYRALIDAE 48 106 24 CRAMBIDAE 134 345 25 DREPANIDAE 24 43 26 LASIOCAMPIDAE 22 61 27 EUPTEROTIDAE 7 20 28 BRAHMAEIDAE 2 2 29 BOMBYCIDAE 6 9 30 SPHINGIDAE 48 168 31 SATURNIIDAE 13 38 32 EPICOPEIIDAE 3 3 33 URANIIDAE 14 43 34 GEOMETRIDAE 269 964 35 NOTODONTIDAE 55 95 36 EREBIDAE 260 868 37 EUTELIIDAE 18 34 38 NOLIDAE 24 57 39 NOCTUIDAE 144 310 TOTAL 1234 3533

Himalayan Lepidoptera Project

Species richness of moths was mapped in 0.15 resolution, i.e. nearly 16 km2 grid cells across the entire Himalayan landscape, to analyse it on a broad geographical basis.

Table 10: Family, Genera and species richness of moths recorded from six Himalayan states in IHR.

State Total No. of Family Total No. of Genus Total No. of Species Jammu & Kashmir 10 79 128 Uttarakhand 31 706 1089 Himachal Pradesh 20 134 370 Sikkim 37 699 2168 West Bengal 17 188 404 Arunachal Pradesh 19 109 235

Potential Sites to repeat Historical Transects Figure 8: Moth richness pattern across Indian Himalayan landscape based on secondary literature

Table 11: Sate-wise and site-wise moth species records from secondary literature. State Sites Geo-Coordinates Species Record Jammu & Kashmir Kashmir 33.7782° N, 76.5762° E 20 Himachal Pradesh Dharamsala 32.2190° N, 76.3234° E 403 Simla 31.1048° N, 77.1734° E 222 Subathu 30.9754° N, 76.9902° E 106 Uttarakhand Bhimtal 29.3461° N, 79.5519° E 579 Kumaon 29.2794° N, 79.4704° E 334 Mussoorie 30.4599° N, 78.0664° E 153 Sikkim Gangtok 27.3389° N, 88.6065° E 28 Tumin 27.3318° N, 88.5142° E 22 West Bengal Darjeeling 27.0360° N, 88.2627° E 1216 Arunachal Pradesh Tawang 27.5781° N, 91.8757° E 105 Siang 28.1097° N, 95.1432° E 92

Himalayan Lepidoptera Project

Objective III: To evaluate status and distribution of threatened Apollo and other Parnassinae butterflies and preparing habitat suitability map to assess optimum habitat conditioning for identification of future suitable habitats for this group under “Climate Envelop Modelling” across Himalayan Biogeographic Provinces.

. Secondary Data Compilation of Apollo Distribution in IHR

Table 12: State-wise records of Apollos from secondary literature. State Total no. of Species Total Number of Number of Exact Recorded Species Entry Locality Recorded Jammu & Kashmir 18 1027 208 Himachal Pradesh 15 138 26 Uttarakhand 8 16 14 Sikkim 16 28 7 West Bengal 1 1 0

. Species Recorded from Primary Field Work So Far

Table 13: Apollo species recorded during the present study. Sl Species Protected Area Locality Altitude Abundance No. (m) 1. Parnassius epaphus Hemis NP, J & K 33.87713 N; 77.62558 E 4237 1

2. Parnassius hardwickii GHNP, HP 31.79999 N; 77.59122 E 2975 3

3. Parnassius hardwickii GHNP, HP 31.72932 N; 77.36654 E 3226 2

4. Parnassius hardwickii AWLS, UK 29.93124 N; 80.38678 E 3218 1

5. Parnassius hardwickii AWLS, UK 29.94033 N; 80.39720 E 3390 1

Parnassius epaphus Parnassius hardwickii

A Typical Habitat of Apollo butterfly sampled within Hemis NP

Himalayan Lepidoptera Project

Objective IV: To compare genetic distinctness of Lepidopteran species commonly occurring in Western, Central and Eastern Himalaya through DNA Barcoding to resolve species complexes and cryptic species, thus generating a barcode database for future molecular and phylogenetic researches on Himalayan Lepidoptera.

. Methodology Adopted

Collection of specimens

Putative morpho-species Morphology data

Species delimitation DNA extraction Estimation of OTUs (Operational Taxonomic Units) PCR, eletrophoresis and Sequencing

Sequence analysis

Genetic divergence Speciation in terms of number of substitution

Diagnostic characters GMYC (The generalised mixed Yule Coalescent) Phylogenetic analysis Reciprocal monophyly

Coalescent analysis Identify independently evolving lineages PTP (Poisson tree process) Evolutionarily significant units (ESUs

 DNA isolation has been carried out for 500 samples using organic phase extraction method/silica column kit method/and magnetic bead based protocol.  Out of 500 samples 468 samples were successfully amplified for about 648 base pairs (bp) from the 5' end of the mtCOI gene using primer pair LCO-HCO. The PCR reaction was set in a 50 µl total volume containing 20 Picomoles of each primer, 20 mM Tris-HCl (pH 8.0), 100 mM KCl, 0.1 mM EDTA,1 mM DTT, 1.8 mM MgCl2, 0.25 mM of each dNTP, and 1U of Taq polymerase (Takara BIO Inc., Japan) with the following cycling parameters: 5 min at 94 °C; followed by 40 cycles of 30 s at 94 °C, 40 s at 49 °C, 1 min at 72 °C and final extension for 5 min at 72 °C.  The amplified PCR products of 468 samples were checked in 1.2% agarose gel. The PCR amplified products were purified using QIAquick Gel Extraction Kit (Qiagen, Valencia, CA) following the manufacturer’s protocols.

Himalayan Lepidoptera Project

 Approximately, 15 ng of the purified PCR product for each of 468 samples was used for sequencing.  The cycle sequencing was performed with BigDye®Terminator ver. 3.1 Cycle Sequencing Kit (Applied Biosystems, Inc.) using 3.2 Picomoles of both forward and reverse PCR primers on a ABI thermal cycler with following parameters: 96 °C for 1 min, then followed by 25 cycles of 96 °C for 10 s, 50 °C for 5 s and a final extension at 60 °C for 1 min15 s.  The cycle sequencing products were cleaned by using BigDye X-terminator kit (Applied Biosystems Inc.) and loaded on 48 capillary ABI 3730 Genetic analyser, housed at Zoological Survey of India, Kolkata.  The SeqScape software version 2.7 (Applied Biosystems Inc.) was used to analyze the forward and reverse chromatograms to obtain the consensus sequences.  The generated forward and reverse sequences were aligned with the representative barcode sequences from global database for similarity search.  The developed sequences were submitted to BOLD database under the project ‘Lepidoptera of Indian Himalayas’ for acquiring the BOLD-IDs.

PCR amplification of partial mitochondrial cytochrome c oxidase subunit I gene segment (about 700 base pairs) of Lepidoptera specimens

Illustration of representative DNA Barcode for Lepidoptera of

Indian Himalayas

Himalayan Lepidoptera Project

Summary of Sequence submission to Barcode of Life Data Systems (BOLD) platform. Annexure III for details.

Himalayan Lepidoptera Project

Plate 3: Different Habitat Types Sampled

Himalayan Lepidoptera Project

Plate 4: Different Habitat Types Sampled

Himalayan Lepidoptera Project

Plate 5

Himalayan Lepidoptera Project

Plate 6

Himalayan Lepidoptera Project

Plate 7

Himalayan Lepidoptera Project

DETAIL OF SEQUENCE SUBMITTED TO BOLD

Himalayan Lepidoptera Project