Comments to be incorporated in the project

1. DNA Barcoading is an ultimate approach towards identification of benthic macro-invertebrates at genome level by using classical taxonomical approach of biomonitoring. 2. Information provided is inadequate in terms of name of moitoring locations and reference site. Comments: Listout the number of sampling locations on River Pamba and specify the reference station located at upstream reach which is not affected by any human influences.

The Ganga basin accounts for a little more than one fourth(26.3 %) of the countrys total geographical area and is the biggest river basin in India, covering the entire states of Uttarakhand , Uttar Pradesh , Bihar , Delhi , parts of Punjab, Haryana, Himachal Pradesh, Rajasthan, Madhya Pradesh and West Bengal. The main river stream originates in the northern most part of Uttarakhand flows through Uttar Pradesh, Bihar and West Bengal and finally drains into the Bay of Bengal. Rising from the icy caves of Gangotri glacier at the height of 4255 above mean sea level, River Ganga starts its long journey to join River Alaknanda and becomes River Ganga near Devprayag. River Ganga is the longest river (2,525 km) and has largest river basin (861,404 km2) in India. The main stretch of River Ganga runs from Haridwar to Allahabad through over Nagal, Bijnor, Garhmukteshwar, Hasanpur, Anupshahar, Narora, Sahaswan, Kasgang, Ptiali, Kampil, Kaimgang, Fatehgarh, Kannauj, Bithaur, Brahmavart, Kanpur and finally Allahabad. At Allahabad, it joins with a major tributary River Yamuna and thereafter passing through Banaras, Patna. At Ganga Sagar in West Bengal, it joins Bay of Bengal. Following sampling locations have been selected on River Ganga. Reference location will be at Gangotri. 1. Gangotri 2. Haridwar 3. Nagal 4. Bijnor 5. Garhmukteshwar 6. Hasanpur

7. Anupshahar 8. Narora

9. Sahaswan 10. Kasgang 11. Ptiali 12. Kampil 13. Kaimgang 14. Fatehgarh 15. Kannauj 16. Bithaur 17. Brahmavart 18. Kanpur 19. Allahabad 20. Banaras 21. Patna 22. Ganga Sagar

3. The details of coverage of reach of the River Ganga, Pamba and tributaries are not specified. Seasonal variation may prevent comparision of sample taken in different seasons. Comments: In the first phase, Main River Pamba and River Ganga will be considered from origin to discharge. The frequency of sampling will be in three different season. For River Ganga, sampling will be carried out during Postmonsoon(Octobr-November), Winter (January-February) and Summer (MayJune). At Reference station sampling will be restricted to Post-monsoon and summer season. 4. The immature stages of mayflies are aquatic. They fly away after attaining mature stages. Adults are delicate insects with a very short life span. They do

not feed(mouthparts are vestigial) and some speciesemerge, reproduce and die in a single daysimultaneously. Benthic macro-invertebrates do not respond to all impacts. Odonate larvae are preyed upon mainly by fish but also by frogs, birds, cray fish, andeach other. In such circumstances, information on mayflies and dragonflies will not be sufficient for life cycle analysis. Comments: The present study does not take into consideration of life cycle analysis. Only the aquatic stages are considered for water quality assessment using Biological Water Quality Criteria developed by CPCB (Table 1). Unlike fish, benthic macro-invertebrates cannot move around much so they are less able to escape from the impacts of sediment and other pollutants that diminishes the water quality. Therefore, benthic macro-invertebrates can give us reliable information on river water quality compared to other aquatic fauna.

Table 1 Biological Water Quality Criteria (BWQC) Sl.No Taxonomic Groups Range of Range of Water quality Water saprobic diversity characteristic quality score Score class (BMWP) 0.2 - 1 Clean A Indicator Colour

Ephemeroptera, Plecoptera, Trichoptera, 7 and more Hemiptera, Diptera Ephemeroptera, Plecoptera, Trichoptera, 67 Hemiptera, Planaria, Odonata, Diptera Ephemeroptera, Plecoptera, Trichoptera, Hemiptera, Odonata, 36 Crustacea, Mollusca, Polychaeta, Diptera Hirudinea, Oligochaeta Mollusca, Hemiptera, Coleoptera, Diptera, 2 5 Oligochaeta Diptera, Oligochaeta No animals 02

Blue

0.5 - 1

Slight Pollution

Light blue

0.3 - 0.9

Moderate Pollution

Green

0.4 Less 0 - 0.2

& Heavy Pollution

Orange

Severe Pollution

Red

5. Despite its relevance, the DNA technique of genetic barcoading will not be able to detect the subtle variations that often delineate very similar specie, sub species and populations from each other. It would be appropriate to associate information for maintenance of aquatic ecology by linking studies on higher forms of fauna present in the reaches of rivers under proposal. Comments: Among all the fresh water invertebrates, only Arthropods, annelids, Molluscs and Platihelminthes are considered for Biological Water Quality assessment wherein taxonomic identification is required up to family/genus/species level. For example, Nymphal stage of Ephemeroptera,Plecoptera and Odonata, Larval stage of Trichoptera, Dipteraand adult stage of Hemiptera, Coleoptera, Molluscs, Annelida and Platihelminthes are living in water bodies. However, insects are confined in River Ganga mostly to the shallow reach upto Allahabado whereas, the deeper reches from Banaras downstream, population of benthic macro-invertebrates are dominated by Crustaceans, Molluscs, Annelids and Platyhelminthes. In view of covering the entire length of River Ganga, all the higher forms of benthic macro-invertebrates may be included in the study. 6. Information about baseline data is not provided in the proposal hence which data set will be used to resolve the phylogenetic relationships among the species is not clear. Comments : Kindly provide your ow overview on the subject. Other comments on the project proposal: Remove all the word such as Imature stages in the text. Do not use samples were identified . Use samples will be identified. From the Materials and methods.