Journal of Threatened Taxa | www.threatenedtaxa.org | 26 April 2019 | 11(6): 13734–13747

 

Baseline biodiversity and physiochemical survey in Parvati Kunda and surrounding area in Rasuwa, Nepal

 

Jessie Anna Moravek 1 , Mohan Bikram Shrestha 2 & Sanjeevani Yonzon 3

 

1,2,3 Wildlife Conservaiton Nepal, Bafal, Kathmandu 44600 Nepal.

1 jessiemoravek@gmail.com (corresponding author), 2 shrmohan5@gmail.com, 3 sanjeevani@wcn.org.np

 

Abstract: Parvati Kunda, a small, alpine wetland located near the village of Gatlang in Rasuwa, Nepal, is a major source of drinking water for the village, possesses spiritual significance, and is a reservoir of local biodiversity.  This study presents the first scientifically conducted biodiversity survey of the wetland.  Here, biodiversity data (wetland plants, birds, mammals, aquatic insects), basic water chemistry (nutrients, pH, dissolved oxygen, conductivity), and basic bacterial tests (total coliform, Escherichia coli, Giardia, Salmonella, Shigella) for the Parvati Kunda wetland is presented from November 2016 and February and May 2017. Parvati Kunda, two of three alternate village water sources, and several village taps were found to be contaminated with E. coli bacteria. Within and around the wetland, 25 species of wetland plants, nine tree species, 10 macroinvertebrate taxa, 37 bird species, and at least six mammal species were documented.  Acorus calamus was the dominant wetland plant and the rapid proliferation of this species over the past twenty years has been reported by community members.  Future studies that further document and monitor wetland biodiversity are necessary.  This study provides a valuable baseline for future research in this culturally and ecologically important wetland.

 

Keywords: Acorus calamus, Escherichia coli, eutrophication, Himalaya, wetland monitoring.

 

doi: https://doi.org/10.11609/jott.4481.11.6.13734-13747   

 

Editor: L.A.K. Singh, Bhubaneswar, Odisha, India.   Date of publication: 26 April 2019 (online & print)

 

Manuscript details: #4481 | Received 11 August 2018 | Final received 01 April 2019 | Finally accepted 05 April 2019

 

Citation: Moravek, J., M.B. Shrestha & S. Yonzon (2019). Baseline biodiversity and physiochemical survey in Parvati Kunda and surrounding area in Rasuwa, Nepal. Journal of Threatened Taxa 11(6): 13734–13747. https://doi.org/10.11609/jott.4481.11.6.13734-13747

 

Copyright: © Moravek et al. 2019. Creative Commons Attribution 4.0 International License.  JoTT allows unrestricted use, reproduction, and distribution of this article in any medium by adequate credit to the author(s) and the source of publication.

 

Funding: The Asia Foundation, The Luce Foundation, Wildlife Conservation Nepal.

 

Competing interests: The authors declare no competing interests.

 

Author details: Jessie Anna Moravek previously worked as a research consultant at Wildlife Conservation Nepal.  She is now a postgraduate student at Lancaster University in the United Kingdom.  Mohan Bikram Shrestha former conservation officer at Wildlife Conservation Nepal.  Sanjeevani Yonzon is the program director at Wildlife Conservation Nepal. 

 

Author contribution: JAM organized and conducted field data collection (plants, trees, macroinvertebrates, water chemistry) and wrote the manuscript.  MBS organized field logistics, conducted field data collection (trees, birds, mammals), and consulted on the manuscript.  SY acted as project supervisor and consulted on the manuscript.

 

Acknowledgements: The authors would like to thank the people of Gatlang, particularly Cher Shing Tamang, Mingmar Tamang, and Nabeen Lama for assistance in logistics.  Further thanks to Rajeshwar Rijal from Wildlife Conservation Nepal, and to Prajjwal Rajbhandari at the Research Institute for Bioscience and Biotechnology.  Dr Ram Devi Tachamo-Shah at Kathmandu University, Dr Nirmala Pradhan at the Nepal Natural History Museum, and persons at the Godawari National Herbarium provided species identification advice.  This work was jointly funded by The Asia Foundation, the Henry Luce Foundation, and Wildlife Conservation Nepal.

 

 

INTRODUCTION

The Himalaya, often considered the “water towers of Asia”, provide water for almost 1.3 billion people living downstream (Xu et al. 2007).  Nowhere, however, are the Himalayan water resources as critical as in the mountains themselves.  Communities throughout the Himalaya depend on alpine wetlands for drinking water, rely on wetland flora and fauna for food and medicine, and attach spiritual significance to bodies of water (Pandit 1999; Xu et al. 2009).  Furthermore, the Himalayan wetlands provide pristine, favourable habitats for flora and fauna in otherwise harsh mountain environments and are therefore important reservoirs of biodiversity (Murray 2009).

The Himalayan wetlands face many threats.  Climate change already causes rapid glacial reduction in many areas and the ongoing rise in temperature will likely impact rain patterns, wetland thermal regimes, and habitat for flora and fauna, jeopardizing sensitive wetland ecosystems and threatening critical water sources (Yao et al. 2004; IPCC 2007; Tse-ring et al. 2010; Gerlitz et al. 2015).  Another major threat to the Himalayan wetlands is eutrophication.  Eutrophication is characterized by excessive plant and algal growth in a body of water, which often results from human activities that produce excessive nitrogen and phosphorus waste, such as raising livestock or applying fertilizer (Carpenter et al. 1998; Schindler 2006; Binzer et al. 2016).  Multiple studies in Himalayan wetlands identified eutrophication from human-generated pollution as a major threat to regional wetland ecosystem health (Khan et al. 2004; Romshoo & Rashid 2014).

Parvati ‘Kunda’ (Nepali: pond; also called ‘Chhodingmo’), a small wetland in the Rasuwa District of Nepal, provides drinking water to the nearby village of Gatlang that consists of about 400 households (Merrey et al. 2018; Fig. 1).  Parvati Kunda also holds spiritual significance to the people in Gatlang and is a harbour of local biodiversity.  A brief, qualitative assessment of Parvati Kunda was performed by World Wildlife Fund in 2007 (Manandhar 2007) and surveys focussed on local livelihoods (e.g., Merrey et al. 2018).  We, however, could not access any scientific studies from Parvati Kunda that may have characterized the wetland and provide data on biodiversity or water chemistry.  As global temperatures rise and human development advances in Gatlang and throughout Nepal, it is critical to establish a baseline in Parvati Kunda now so as to assess the extent and consequences of future changes.  Furthermore, such baseline information will characterize the contribution of Parvati Kunda to regional vegetation and bird and macroinvertebrate biodiversity, and ultimately provide information that will promote the sustainable management of the wetland ecosystem.

 

Study site

Parvati Kunda is in ward number three of Aama Chhodingmo Rural Municipality, about 100km north of Kathmandu in Nepal’s Rasuwa District.  Climactically, the region is cold and snowy in December through February and temperature maximums occur between May and July.  Seasonal climate variations are dominated by the Indian monsoon that occurs between June and September, while small scale climactic variations depend on altitude and aspect (Kharel 1997).

Parvati Kunda is just outside Langtang National Park.  The NP encompasses 1,710km2 (660mi2) area in Nuwakot, Rasuwa, and Sindhulpalchok districts in the central Himalayan region.  Langtang NP and the surrounding area are home to 46 types of mammals and 250 species of birds, including several rare and protected species such as Snow Leopard Panthera uncia, Clouded Leopard Neofelis nebulosi, Leopard Panthera pardus, and Red Panda Ailurus fulgens (Fox et al. 1996; DNPWC 2019).

Gatlang Village is located at 2,300m on a trekking route called Tamang Heritage Trail and is known for its traditional Tamang architecture.  Most inhabitants of Gatlang identify as Tamang and speak the Tamang language (Merrey et al. 2018).  Like other parts of the Langtang region, people grow potato, maize, buckwheat, beans, millet, and barley; raise ‘chauri’ (Nepali: yak-cow hybrid), sheep, and goats; and run lodges, jeep businesses, or make handicrafts that cater to tourists (Merrey et al. 2018).  Gatlang is accessible by road, although there is no regular bus service.

Parvati Kunda (85.261oE & 28.156oN) is a 1.87---hectare groundwater and rainfall/ snowmelt-fed eutrophic wetland located at 2,600m.  The wetland is located 300m above Gatlang and lies on the northern edge of Bongjomane Community Forest from which the community members harvest fodder, firewood, and timber.  Parvati Kunda is religiously significant and is often the subject of local religious festivals.  Water from the deep centre of the wetland is connected to a system of pipes and public taps in the village.  Some village taps are fed from one of the three additional springs in the vicinity.  Water flows out through a small opening on the northeastern edge of the wetland, where a man-made wall confines the outflow channel to about 1m in width (Fig. 3a).  The outflow descends 300m over 1km to Gatlang Village.

 

METHODS

Parvati Kunda was surveyed three times, in November 2016 and in February and May 2017.  Surveys for particular taxa groups were conducted when seasonally appropriate.  Vegetation and physicochemical parameters were measured in 18 quadrats within the wetland, and others were analyzed in a subset of 10 for macroinvertebrates and 12 nutrient quadrats. 

 

Wetland Vegetation

The locations of vegetation quadrats within Parvati Kunda included the inflow plus 17 randomly determined locations from ArcGIS (ESRI 2014; Fig. 3b).  Vegetation surveys included areas that were permanently inundated with emergent or floating macrophytic vegetation, but excluded open water, i.e., the areas inundated but lacking emergent macrophytic vegetation.  Open water was excluded from the location generator using satellite imagery and ArcGIS (ESRI 2014; Google Earth 2016).  No survey site was more than 0.7m deep and all could be accessed by wading.  Vegetation surveys were conducted in November and May only.  February was excluded because there was minimal vegetation growth.  Quadrat-based sampling was applied to the vegetation study using 0.5m2 square quadrats and plants were identified using standard taxonomic keys (Malla et al. 1976).  Unidentified species were collected in herbarium sheets for identification by experts at the National Herbarium in Godavari.  Per cent cover of each species was visually estimated and the number of individuals of each species was counted.  For rhizomous species such as Juncus leucanthus or Acorus calamus, where multiple shoots might be connected by a single root system, individual shoots were counted to avoid digging up the entire plant.  In cases where moss was present, the number of individuals were counted in three 10cm x 10cm sample plots and used to calculate the average number of individuals per square metre.  This was used to estimate the number of individuals in quadrats.

Specific information about the proliferation of A. calamus in Parvati Kunda over the last 20 years was gathered during conversations with community members.  Such local accounts are considered important to understand changes in Parvati Kunda over time since prior published data on wetland vegetation does not exist to the best of our knowledge.  Before commencing biodiversity surveys, the researchers interviewed seven high-profile community leaders specifically about water resources in the village.  Further insight into questions about changes in Parvati Kunda could be obtained from 31 household surveys that were conducted in conjunction with another study during this time.

 

Riparian Trees

Four circular riparian tree plots were created on the northern, southwestern, southern, and northeastern sides of the wetland (Fig. 3c).  The plots measured 5m in radius, 78.54m2 in area, covering at least 5% of the riparian area as defined by the wetland boundary wall.  Trees with a diameter at breast height greater than 10cm were identified and counted.

 

Aquatic Macroinvertebrate Survey

Aquatic macroinvertebrate surveys were conducted in May in 10 of the vegetation plots using the pipe method (Britton & Greeson 1989), where a sturdy plastic bucket with a height of 0.81m and diameter 0.288m and no bottom was placed firmly in the mud at each sampling location (Fig. 3d).  Using a net, the water inside the bucket was agitated and all insects and loose debris were scooped out with five to six scoops of the net, ensuring that at least three scoops scraped the bottom.  Contents of the net were placed in a sieve and excess debris was rinsed and removed in the field.  The remaining debris and macroinvertebrates were preserved in 60% ethanol.  In the laboratory, invertebrates were sorted, counted, and identified to family level.  The Nepal lake biotic index (NLBI) and lake water quality class (LWQC) were calculated using family level identification according to Tachamo-Shah et al. (2011). 

 

Bird Survey

In November, February, and May the point-count method was used for bird observations in which a single observer stayed in one fixed position during a specified period of time and recorded all birds seen or heard during that period (Ralph et al. 1995).  This method was used for one hour in the morning and evening.  The timings of observations were 07.00–08.00 h and 16.00–17.00 h in May and 08.00–09.00 h and 15.00–16.00 h in November and February, for two consecutive days during each survey period.  A single observer stood on the northern bank of the wetland where they had a good view of the entire wetland area with the aid of binoculars.

 

Wildlife Observation

Data on animals were gathered from direct sighting, evidence of animal usage from faeces, wallows and walking signs, and mentions by local people.  Photographic evidence of animals were collected when possible.

 

Physiochemical Parameters

Physical water parameters were measured in each of 18 quadrats and the outlet in November, February, and May.  Measurements for dissolved oxygen, temperature, pH, and electrical conductivity were performed using handheld probes, namely, Lutron DO meter 519, Hanna pHep meter, and Hanna DiST meter.  Nitrate, ammonia, and total phosphate were measured at a subset of 12 quadrats (Fig. 3e).  To measure nutrients, samples were collected in 500ml clean plastic sample bottles, kept cold, and transported to Kathmandu within 24 hours.  Lab work and analysis were performed by CEMAT Water Labs, Kathmandu, Nepal, the procedural references for which are presented in Table 1.

 

Biological Water Quality

Because Parvati Kunda is an important source of drinking water for the Gatlang community, we performed bacterial tests in the wetland and in several other spring-fed water sources and village taps.  Using public data on water-borne illnesses from the village health centre to guide our study, we tested for multiple bacterial contaminants, including total coliform, Escherichia coli, Salmonella, Shigella, Giardia, and for the possible presence of any ova, worms, cysts, or amoeba.  Tests for total coliform, E. coli, Salmonella, and Shigella were performed in February and May.  Tests for Giardia and other microscopic stages were done in May.  Sampling was performed at three locations in Parvati Kunda—the inlet, the outlet, and WQ1.  Tests were also conducted in three additional spring sources, namely, Chyange Spring, Sanglang Ghode Spring, and Shernemba Spring, which supplied water to six village taps (Fig. 4).  Water samples were collected in pre-sterilized 250ml collection bottles, kept cold, and transported to Kathmandu within 24 hours.  Lab work and analysis was performed by CEMAT Water Labs (Table 1).

 

Data Analysis

For analysis of wetland vegetation and data from tree surveys, we used density and Simpson’s reciprocal index of biodiversity,

 

 

where n = the total number of organisms of a particular species and N = the total number of organisms of all species.  The indices were calculated for each quadrat using the vegan package in R (R Development Core Team 2016).  Simpson’s reciprocal index was used because it tends to be less sensitive to both the shape of the abundance distribution and small sample sizes, compared to other diversity metrics (Gotelli & Chao 2013).

NLBI was calculated by assigning scores to each macroinvertebrate identified according to family.  The sum of these scores divided by the number of scored taxa is equivalent to the NLBI.  The NLBI score was then translated to a LWQC rating that indicates the degree of pollution likely in the waterbody (Tachamo-Shah et al. 2011).

Total phosphorus was determined using the ammonium molybdate spectrometric method.  Ascorbic acid and acid molybdate were added to a 30ml filtered subsample and formed a complex with orthophosphate ions.  The complex was then reacted with ascorbic acid to form a blue compound, and the absorbance of this compound was measured against a calibration curve to determine the concentration of orthophosphate in the sample (ISO 6878:1998(E)).  Nitrate was measured with the spectrometric method using sulfosalicylic acid.  Sodium salicylate and sulfuric acid were added to a 30ml filtered subsample and reacted with nitrate.  After alkali treatment, a yellow compound formed.  The absorbance of this compound was used to determine the nitrate concentration (ISO 7890-3).  The nesslerization method was used to measure ammonia concentration.  The water sample was filtered, buffered with a borate buffer, and then distilled into a boric acid solution.  Nessler reagent was added to the distillate, reacted with ammonia to produce a yellow colour, and ammonia concentration was measured spectrophotometrically (4500-NH3 C, APHA 17th Ed.).

Total coliform was identified using a fermentation technique.  Lauryl tryptose broth was inoculated in fermentation tubes and incubated for 24h at 35⁰C.  If gas or acid was produced, samples were transferred to green lactose bile and incubated for 48h at 35⁰C.  If coliform colonies were observed, samples were transferred to an agar slant and lauryl tryptose broth fermentation tube for 24h at 35⁰C.  If gas was produced at this stage, part of the agar slant growth was gram stained.  If gram negative rods were present, coliform was present.  Coliform density was estimated by calculating the most probable number (MPN) value from the number of positive green lactose bile tubes (9221 B, APHA 17th Ed.).  Escherichia coli was identified using the membrane filter method.  A quantity of 100mL of sample water was filtered through a 0.45μmpore size membrane filter and placed on a plate of M-endo agar.  The plate was incubated at 35⁰C for 24h and bacterial colonies were inspected for red-metallic colour indicating E. coli.  Density was estimated with MPN (9221 F, APHA 17th Ed.).

Salmonella and Shigella samples were pre-enriched in peptone water, then enriched in tetrathionate broth and incubated at 35⁰C for 24h.  The samples were streaked on Salmonella Shigella (SS) agar.  The selective solid media allowed plates to be screened for the presence or absence of both Salmonella and Shigella (9260 B and E, APHA 17th Ed.).  Giardia, ova, worms, and cysts were identified by microscopic observation of 100ml of sample for presence/ absence analysis.           

 

RESULTS

Biodiversity Surveys

Vegetation

Twenty-five species of plants were identified from surveys of wetland vegetation (Fig. 3b; Table 2).  Surveys also identified a large population of Sphagnum palustre, a type of peat moss that is rare in Rasuwa District (Nirmala Pradhan pers. comm. December 2016).

The total area of the wetland dominated by Acorus calamus was demarcated by a combination of field measurements on a geographic positioning system (GPS) and visual analysis of satellite imagery in ArcGIS (ESRI 2014; Fig. 3a).  Acorus calamus covered about 7,140m2 of the wetland or 38% of the total wetland surface area.  Within vegetation survey plots, A. calamus covered an average of 37% of the area within the 18 survey quadrats, verifying the visual determination of A. calamus coverage in ArcGIS.  

Five of seven key informants identified an increase in A. calamus and perceived the species as a major threat to the wetland.  Sixty-five per cent of respondents in household surveys also identified an increase in A. calamus as the most noticeable change in Parvati Kunda over the past 20 years (Yonzon unpub. data).

Nine species of trees were identified during tree surveys in four different survey plots around the wetland (Fig. 2; Table 3).  Tree diversity (Simpson’s reciprocal index) ranged from 2.571 on the western side (plot 2) to 4.378 on the eastern side (plot 4) of the wetland.

 

Macroinvertebrate Surveys

In total, 10 aquatic macroinvertebrate taxa were identified through surveys (Table 4).  The NLBI was calculated using these invertebrates to assess the overall ecological quality of the wetland.  Parvati Kunda received a ‘poor’ LWQC rating, indicating a likely high level of nutrient pollution (Tachamo-Shah et al. 2011).

 

Bird and Mammal Surveys

Point-count bird surveys identified 37 species of birds in and around the wetland (Table 5).  Opportunistic observations by the field research team confirmed the presence of six mammal species in or near the wetland area, within 10m from the water’s edge.  Community members identified four additional mammals that were not directly observed by the researchers (Table 6).

 

Water Properties

Ammonium and total phosphate levels were mostly negligible (<0.1mg/l).  Nitrate was higher at the inlet (2.3mg/l in February and 2.9mg/l in May), but low or negligible in other locations (Table 7).  Other parameters including temperature, dissolved oxygen, pH, and electrical conductivity differed over seasons.  Average values are presented in Table 8.

 

Biological Contaminant Tests

Parvati Kunda was found to be contaminated with E. coli in both February and May and the Giardia trophozoite that is an active stage was found in May.  Two out of the three local water sources, namely, Chyange Spring and Shernemba Spring, were contaminated with E. coli, Giardia, Salmonella, Shigella, and ova.  Compared to water sources, taps generally contained higher levels of E. coli and greater variation in other tested contaminants (Table 9).

 

DISCUSSION

Change in Vegetation Profile

Based on biodiversity surveys, the semi-aquatic plant Acorus calamus was by far the most dominant plant species in the wetland.  Sphagnum palustre is the second most dominant species and is an important structural plant since it influences water chemistry and provides a platform for other plants to grow on (Glime et al. 1982).  According to accounts obtained from community members during casual conversation as well as interviews of key informants, the proliferation of A. calamus in Parvati Kunda over the past 20 years was high and is a concern for the wetland ecosystem as a source of water for people, animals, and plants. 

Monitoring changes in Parvati Kunda vegetation such as A. calamus is important because the species composition of wetland macrophytes often reflects water quality.  For example, one study in lake Hiidenvesi in Finland identified notable changes in vegetation communities as lake eutrophication progressed—with vegetation changing from species requiring coarse bottom material and high light levels to species favouring soft bottom substrate and turbid waters (Nurminen 2003).  Similarly, a study in the Tiber River basin in Italy identified a certain selection of wetland plants as bioindicators of water pollution and eutrophication (Ceschin et al. 2010).  Acorus calamus, in particular, was identified as especially tolerant to high nitrogen levels and anoxia (Weber & Brändle 1996; Zhang et al. 2018) and therefore the alleged increase in dominance of this species over time in Parvati Kunda could be an indicator of high nitrogen levels and anoxia in the wetland.  A study in the Hokersar Wetland in the Kashmir Himalaya in India, however, found that A. calamus nearly disappeared in the wetland, which was affected by pollution, siltation, and agriculture and had high levels of nitrogen (Khan et al. 2004).  It is not clear exactly what differences between Hokersar and Parvati Kunda wetlands led to different responses in the A. calamus population, but this could be investigated with further study.  

Understanding the proliferation of A. calamus in Parvati Kunda is also important from the biodiversity perspective.  The changes in water quality and associated changes in wetland vegetation can lead to some plant species out-competing others.  For example, a long-term study in wetlands in the Jura Mountains in Switzerland found that due to N-deposition, species composition trended towards nutrient-rich flora and some species were out-competed and disappeared from the ecosystem over a period of 25 years (Rion et al. 2018).  A similar trend was found at lake Hiidenvesi in Finland, where certain species requiring hard substrates were out-competed by those preferring soft substrates (Nurminen 2003).  It is plausible that the proliferation of A. calamus identified by local community members could have similar effects on wetland plant diversity and community composition.  Although our study did not examine the specific biodiversity impacts of A. calamus, future studies in Parvati Kunda should make use of the baseline information presenting possible threats here to understand the consequences of A. calamus growth within the wetland ecosystem.  

Given the dominance of A. calamus in Parvati Kunda, we expected to find high levels of nutrients in water samples.  This was not the case and nutrient levels were, in most cases, negligible.  Since A. calamus grows both in water and on wetland edges or areas that are not permanently inundated, however, soil could be another critical source of excess nutrients.  For example, a study on residual availability of nitrogen in soil following application of organic fertilizer from farmyard manure or compost found that nitrogen can be immobilized in soils and enrich the soil nitrogen pool over a long term (Gutser et al. 2005).  A similar phenomenon may happen near Parvati Kunda, leading to slow release of nitrogen from the soil rather than from the water.  Furthermore, nutrient addition studies on invasive Phragmites australis in the United States found that nitrogen added to soil, rather than water, significantly increased the growth of the wetland plant P. australis compared to the native Spartina pectinata (Rickey & Anderson 2004).  This indicates that nutrient enrichment of soil can indeed increase growth rates of wetland plants.  Additionally, A. calamus itself might absorb much of the nitrogen entering the wetland.  Zhang et al. (2018) investigated the nitrite stress tolerance of A. calamus during wastewater treatment and found that A. calamus could tolerate up to 30mg/l of nitrite (2018).  Likewise, a study in northeastern China identified A. calamus as a tool for nitrogen removal in floating treatment wetlands, finding that A. calamus demonstrated an 84.2% removal efficiency for total nitrogen (Li & Guo 2017).  Thus, ensuing studies on A. calamus in Parvati Kunda should focus on the nutrient content of both A. calamus and the soil to fully understand the nutrient dynamics in the wetland.

 

Nutrient pollution and microorganisms

The presence of excess nutrients in Parvati Kunda is also indicated by the ‘poor’ LWQC score, which suggests a ‘heavy’ degree of nutrient pollution (Tachamo-Shah et al. 2011).  This rating is related to the diversity and types of macroinvertebrates present in the wetland.  Several direct explanations for a poor rating may exist; for example, lack of substrate and lack of dissolved oxygen as a result of excess nutrient inputs and eutrophication processes can inhibit macroinvertebrate breeding, emergence, and overall survival (Boles 1981; Connolly et al. 2004).  When abiotic conditions such as nutrient levels, substrate type, and dissolved oxygen are unfavourable, the rest of the ecosystem is affected.  It should be noted, however, that although Parvati Kunda received a poor rating, the pipe method used for macroinvertebrate sampling was quantitative instead of qualitative as the NLBI method suggests (Tachamo-Shah et al. 2011).  Even if a more extensive qualitative survey were conducted, however, the LWCQ rating would most likely only move one step from poor to fair.  This would still indicate a moderate level of nutrient pollution.  Furthermore, wetlands in monsoonal systems such as in Nepal are known to change in water quality ranking depending on the season.  For example, a study in the Rampur Ghol ecosystem in Chitwan, Nepal, used a similar macroinvertebrate indicator system to describe water quality and found that water quality and macroinvertebrate diversity was generally lower in the dry season than the rainy season (Gautam et al. 2014).  Likewise, the LWCQ rating in Parvati Kunda probably fluctuates throughout the year and additional qualitative sampling would possibly identify either poor or fair ratings depending on the season.

Several of the macroinvertebrate taxa identified in this study indicate specific details about Parvati Kunda and its level of pollution.  For example, many prior studies found that Chironomidae abundance generally increases with decreasing habitat quality (Fore et al. 1996; Botts 1997) and red Chironomidae are easily identifiable and are therefore a good indicator of this pattern (Tachamo-Shah et al. 2011).  Red Chironomidae was very abundant at all sampling locations within Parvati Kunda, indicating high levels of pollution.  Furthermore, various species in Insecta families of Heteroptera and Corixidae were used as indicators of eutrophication and pollution in Finland (Jansson 1977).  As the Finland study found, species within the Corixidae family vary greatly in their tolerance to pollution, they were assigned a low rating (2) in the NLBI, and their presence in Parvati Kunda indicates high levels of nutrient pollution that may be specifically linked to eutrophication.

 

Threat of Eutrophication

Although wetland eutrophication is a natural process, it is unclear whether human activities within the Parvati Kunda watershed may also be influencing degradation of the wetland.  Activities such as agriculture, livestock raising, and timber harvesting within the wetland watershed may lead to higher levels of nutrient inputs that could explain the patterns we observed in Parvati Kunda (Foote et al. 1996; Prasad et al. 2002).  Most of the Kunda watershed is encompassed by Bongjomane Community Forest, which is utilized by Gatlang community members for fodder and timber harvesting.  Livestock also inhabits the area around the wetland. Future studies should use GIS and satellite imageries to assess landuse changes in the Parvati Kunda watershed over the last 30 years.  Such a study could help indicate how and why nutrient pollution occurred in the wetland.

Public records available at the village health centre showed persistent infections of intestinal worms, Salmonellosis, and Shigellosis, and we indeed found associated contaminants in Parvati Kunda, three alternative water sources, and several village taps.  Of all water sources and taps tested, only one water source at Sanglang Ghode spring was in compliance of Nepal national drinking water quality standards (Ministry of Physical Planning and Works 2005).  Livestock waste is a probable source of contamination in the wetland itself, especially since livestock was frequently observed inside the wetland enclosure and was observed defecating near the water during bacterial sampling.  Other reasons for contamination may include leaking pipes through which contaminants can seep between the water source and village taps.  The extent and source of bacterial contamination in the wetland, other water sources, pipes, and taps should be further investigated, and methods for maintaining acceptable drinking water quality in the village should be pursued.

 

CONCLUSION

The present study is a baseline study in the Parvati Kunda Wetland and is believed to provide the first description of its wetland vegetation, riparian trees, aquatic macroinvertebrates, birds, physiochemical water parameters, and biological contaminants.  Our data indicate a moderate to heavy degree of nutrient pollution, although the direct and indirect causes and consequences of pollution are not fully understood.  Continued monitoring of the Parvati Kunda wetland will provide a valuable and interesting example of the process of wetland degradation and conservation in the Nepal Himalaya.

Future studies should focus on amphibian- and mammal-specific surveys.  Additionally, watershed level studies that describe land-use changes and may indicate reasons for wetland degradation are necessary.

 

 

Table 1. Tests and methods used for detection and quantification of nutrients and detection of pathogens. 
All tests were performed by CEMAT Water Labs in Kathmandu, Nepal.

 

Test

Method used

Total phosphorus

ISO 6878:1998(E)

Nitrate

ISO 7890-3

Ammonia

4500-NH3 C, APHA 17th Ed.

Total coliform

9221 B, APHA 17th Ed.

E. coli

9221 F, APHA 17th Ed.

Salmonella spp.

9260 B, APHA 17th Ed.

Shigella spp.

9260 E, APHA 17th Ed.

Ova/ worms/ Giardia/ amoeba/ cyst

-

 

 

Table 2. Wetland vegetation recorded in Parvati Kunda, Nepal, in 2016–2017.

 

Order

Family

Scientific name

Acorales

Acoraceae

Acorus calamus

Apiales

Apiaceae

Acronema tenerum

Asparagales

Orchidaceae

Spiranthes sinensis

Asterales

Asteraceae

Aster tricephalus

Asterales

Asteraceae

Pseudognaphalium affine

Asterales

Asteraceae

Senecio laetus

Brassicales

Brassicaceae

Cardamine flexuosa

Brassicales

Brassicaceae

C. impatiens

Brassicales

Brassicaceae

C. loxostemonoides

Caryophyllales

Caryophyllaceae

Arenaria debilis

Caryophyllales

Polygonaceae

Persicaria nepalensis

Caryophyllales

Polygonaceae

Persicaria sp.

Caryophyllales

Polygonaceae

Rumex nepalensis

Lamiales

Lamiaceae

Elsholtzia strobilifera

Lamiales

Scrophulariaceae

Hemiphragma heterophyllum

Lycopodiales

Lycopodiaceae

Lycopodium japonica

Malpighiales

Hypericaceae

Hypericum japonica

Paeoniales

Paeoniaceae

Rubus terutleri

Poales

Juncaceae

Juncus himalensis

Poales

Juncaceae

J. leucanthus

Poales

Poaceae

Calamogrostis pseudophragmites 

Poales

Poaceae

Miscanthus nepalensis

Ranunculales

Ranunculaceae

Anemone rivularis

Ranunculales

Ranunculaceae

Ranunculus cantoniensis

Sphagnales

Sphagnaceae

Sphagnum palustre

 

 

Table 3. Trees found in riparian areas around Parvati Kunda, Nepal, in 2016–2017.

 

Order

Family

Scientific name

Common name

Plot 1

Plot 2

Plot 3

Plot 4

Ericales

Ericaceae

Rhododendron barbatum

Rhododendron

5

 

 

 

Ericales

Ericaceae

Lyonia ovalifolia

Angeri

 

4

1

2

Fagales

Fagaceae

Quercus semicarpifolia

Oak

6

 

1

7

Laurales

Lauraceae

Cinnamomum spp.

Wild Cinnamon

6

2

2

2

Laurales

Lauraceae

Persea odoratissima

Pra

1

 

 

 

Laurales

Lauraceae

Cinnamomum glanduliferum

Wild Cinnamon

 

 

 

2

Malpighiales

Salicaceae

Xylosma controversum

Willow

 

 

6

2

Pinales

Pinaceae

Tsuga dumosa

Himalayan Hemlock

 

 

 

3

Sapindales

Sapindaceae

Acer spp.

Maple

 

6

3

 

Biodiversity index

3.306

2.571

3.267

4.378

 

 

Table 4. Aquatic macroinvertebrates found in Parvati Kunda, Nepal, in 2016–2017.  Two taxa are repeated twice (Haplotaxida: Tubificidae and Coleoptera: Dytiscidae).  They represent different species.  Identification up to the level of family was needed for NLBI calculation and family was only scored once in NLBI.  Trichoptera: Polycentropodidae is not included in the NLBI index score.

 

 

Order

Family

NLBI Score

1

Diptera

Chironomidae

1

2

Haplotaxida

Tubificidae

2

3

Haplotaxida

Tubificidae

Replicate

4

Trichoptera

Polycentropodidae

NA

5

Coleoptera

Dytiscidae

5

6

Odonata

Libellulidae

3

7

Coleoptera

Dytiscidae

Replicate

8

Hemiptera

Corixidae

2

9

Hemiptera

Gerridae

4

10

Veneroida

Sphaeriidae

5

NDBI score/ LWQC rating

3.14/ poor

 

 

Table 5. Bird species of Parvati Kunda and surrounding areas, Nepal.  LC = Least Concern according to IUCN (2016).

 

Order

Family

Scientific name

Common name

IUCN status

Accipitriformes

Accipitridae

Spilornis cheela

Crested Serpent-eagle

LC

Columbiformes

Columbidae

Streptopelia orientalis

Oriental Turtle Dove

LC

Cuculiformes

Cuculidae

Cuculus canorus

Common Cuckoo

LC

Cuculiformes

Cuculidae

C. saturatus

Oriental Cuckoo

LC

Galliformes

Phasianidae

Francolinus francolinus

Black Francolin

LC

Gruiformes

Rallidae

Zapornia bicolor

Black Tailed Crake

LC

Passeriformes

Aegithalidae

Aegithalos concinnus

Black-throated Tit

LC

Passeriformes

Cettiidae

Horornis flavolivacea

Aberrant Bush Warbler

LC

Passeriformes

Corvidae

Corvus macrorhynchos

Large-billed Crow

LC

Passeriformes

Corvidae

Pyrrhocorax pyrrhocorax

Red-billed Chough

LC

Passeriformes

Corvidae

Urocissa flavirostris

Yellow-billed Blue Magpie

LC

Passeriformes

Dicruridae

Dicrurus leucophaeus

Ashy Drongo

LC

Passeriformes

Fringillidae

Carduelis spinoides

Yellow-breasted Greenfinch

LC

Passeriformes

Fringillidae

Carpodacus thura

White-browed Rosefinch

LC

Passeriformes

Laniidae

Lanius schach

Long-tailed Shrike

LC

Passeriformes

Laniidae

L. tephronotus

Grey-backed Shrike

LC

Passeriformes

Leiothrichidae

Trochalopteron lineatum

Streaked Laughingthrush

LC

Passeriformes

Leiothrichidae

T. variegatum

Variegated Laughingthrush

LC

Passeriformes

Leiothrichidae

Heterophasia capistrata

Rufous Sibia

LC

Passeriformes

Motacillidae

Anthus spp.

Pipit spp.

 -

Passeriformes

Motacillidae

Motacilla cinerea

Grey Wagtail

LC

Passeriformes

Muscicapidae

Enicurus maculatus

Spotted Forktail

LC

Passeriformes

Muscicapidae

Ficedula westermanni

Little Pied Flycatcher

LC

Passeriformes

Muscicapidae

Myophonus caeruleus

Blue Whistling-thrush

LC

Passeriformes

Muscicapidae

Phoenicurus frontalis

Blue-fronted Redstart

LC

Passeriformes

Muscicapidae

Saxicola ferreus

Grey Bushchat

LC

Passeriformes

Muscicapidae

S. torquatus

Common Stonechat

LC

Passeriformes

Paridae

Lophophanes dichrous

Grey-crested Tit

LC

Passeriformes

Paridae

Parus monticolus

Green-backed Tit

LC

Passeriformes

Pycnonotidae

Pycnonotus leucogenys

Himalayan Bulbul

LC

Passeriformes

Sittidae

Sitta himalayensis

White-tailed Nuthatch

LC

Passeriformes

Stenostiridae

Culicicapa ceylonensis

Grey-headed Canary-flycatcher

LC

Passeriformes

Sylviidae

Fulvetta vinipectus

White-browed Fulvetta

LC

Passeriformes

Timaliidae

Pomatorhinus erythrogenys

Rusty-cheeked Scimitar-babbler

LC

Passeriformes

Troglodytidae

Troglodytes troglodytes

Northern Wren

LC

Passeriformes

Turdidae

Turdus unicolor

Tickell's Thrush

LC

Piciformes

Picidae

Picus squamatus

Scaly-bellied Woodpecker

LC

 

 

Table 6. Mammal species occurring around Parvati Kunda, Nepal, in 2016–2017. 
LC = Least Concern and V = Vulnerable according to IUCN (2018).

 

Taxa

Scientific name

Direct observation

Secondary information

IUCN status

Nepal Grey Langur

Semnopithecus schistaceus

x

 

LC

Barking Deer

Muntiacus muntjak

x

 

LC

Royles Pika

Ochotona roylei

x

 

LC

Yellow-throated Marten

Martes flavigula

x

 

LC

Chauri

-

x

 

-

Goats/ Sheep

-

x

 

-

Porquipine

Hystrix spp.

 

x

-

Leopard

Panthera pardus

 

x

V

Mongoose

Herpestes spp.

 

x

-

Wild Boar

Sus scrofa

 

x

LC

 

 

Table 7. Total phosphorus, nitrate, and ammonia measurements for February (10 wetland locations, see Fig. 3e) and
May (three wetland locations—outlet, inlet, and WQ3).  Fields in this table represent a single data point;
only one sample was taken in each location in each season.

 

Sample location

T. phosphorus (mg/l as P)

Nitrate (mg/l as NO3)

Ammonia (mg/l as NH3)

 

February

May

February

May

February

May

Outlet

<0.1

<0.1

<0.1

0.3

<0.1

<0.1

Inlet

<0.1

<0.1

2.3

2.9

<0.1

<0.1

WQ1

<0.1

 

0.7

 

<0.1

 

WQ2

<0.1

 

0.4

 

<0.1

 

WQ3

<0.1

<0.1

<0.1

0.4

<0.1

0.8

WQ4

<0.1

 

<0.1

 

<0.1

 

V3

<0.1

 

0.1

 

<0.1

 

V5

<0.1

 

<0.1

 

<0.1

 

V6

<0.1

 

0.5

 

0.1

 

V8

<0.1

 

<0.1

 

<0.1

 

V9

<0.1

 

<0.1

 

<0.1

 

V10

<0.1

 

0.1

 

<0.1

 

V11

<0.1

 

0.7

 

<0.1

 

 

Table 8. Temperature, dissolved oxygen, pH, and electrical conductivity in 18 sampling locations in the wetland. 
Averages and standard deviations (SD) from the three sample points (November 2016 and February and May 2017) are presented. 
Note: pH was measured only in February and May and conductivity was measured only in November and February.

 

Sample location

Temperature (⁰C)

Dissolved oxygen (mg/l)

pH

Conductivity (mg/l)

Average

SD

Average

SD

Average

SD

Average

SD

Outlet

13.0

2.6

7.4

0.6

7.0

0.5

30.0

0.0

Inlet

12.2

0.6

7.3

0.5

6.2

0.3

20.0

0.0

WQ1

15.5

4.3

5.8

1.2

6.8

0.4

30.0

0.0

WQ2

14.7

6.3

5.5

3.1

5.9

1.0

20.0

0.0

WQ3

20.9

4.1

9.0

6.5

7.0

1.1

20.0

10.0

WQ4

13.9

8.8

8.4

0.7

6.1

0.4

20.0

0.0

WT

15.8

3.5

2.1

1.2

4.5

0.3

10.0

0.0

V1

13.7

2.1

9.4

0.9

6.8

0.4

30.0

0.0

V2

18.0

0.7

10.8

5.8

6.4

1.1

20.0

0.0

V3

16.7

7.0

5.3

2.9

5.9

0.8

30.0

10.0

V4

12.7

4.0

5.5

1.7

5.7

1.1

20.0

0.0

V5

13.4

6.9

6.0

3.2

6.6

0.6

25.0

5.0

V6

14.2

7.7

7.9

1.1

6.4

0.4

20.0

0.0

V7

18.0

10.1

6.7

2.3

6.6

0.2

35.0

5.0

V8

14.0

8.6

7.6

1.2

6.8

0.2

30.0

0.0

V9

14.9

10.2

7.9

0.9

7.0

0.5

30.0

0.0

V10

12.6

9.0

5.6

1.6

5.7

0.5

5.0

5.0

V11

12.8

3.5

5.3

0.3

6.7

0.8

40.0

0.0

V12

10.0

6.6

5.4

1.8

5.9

0.9

20.0

0.0

 

 

Table 9.  Results of tests conducted in February and May 2017 for presence (P) or absence (A) of microorganism per 100ml of water samples from 12 different locations near Parvati Kunda, Nepal.  MPN Index stands for most probable number of colonies present in 100ml of sample, which is determined by comparing the pattern of positive results with statistical tables (Bartram & Pedley 1996).  Key to sample locations (Fig. 4): 1 - PK Inlet, 2 - PK Outlet, 3 - PK WQ1, 4 - Chyange Spring, 5 - Chyange Tap, 6 - Sanglang Ghode Spring, 7 - Sanglang Ghode Tap, 8 - Shernemba Spring, 9 - Shernemba Pipe, 10 - Mill Tap, 11 - Old Parvati Kunda Tap, 12 - New Parvati Kunda Tap.

 

Sample location

Total coliform (MPN index/100ml)

E. coli (MPN index/100ml)

Salmonella spp. (P/A)*

Shigella spp. (P/A)*

Ova (P/A)*

Worms (P/A)*

Giardia (P/A)*

Amoeba (P/A)*

Cyst (P/A)*

 

Feb

May

Feb

May

Feb

May

Feb

May

Feb

May

Feb

May

Feb

May

Feb

May

Feb

May

1

9

150

4

9

-

A

-

A

-

A

-

A

-

A

-

A

-

A

2

0

1100

0

460

A

A

A

A

-

A

-

A

-

A

-

A

-

A

3

-

210

-

39

-

A

-

A

-

A

-

A

-

P

-

A

-

A

4

93

23

43

4

P

A

P

P

-

P

-

A

-

A

-

A

-

A

5

-

4

-

4

-

A

-

A

-

A

-

A

-

A

-

A

-

A

6

-

0

-

0

-

A

-

A

-

A

-

A

-

A

-

A

-

A

7

-

1100

-

43

-

A

-

P

-

A

-

A

-

A

-

P

-

A

8

-

240

-

43

-

A

-

A

-

A

-

A

-

A

-

A

-

A

9

-

240

-

43

-

A

-

A

-

A

-

A

-

A

-

A

-

P

10

460

1100

240

240

P

A

P

A

-

A

-

A

-

A

-

P

-

A

11

43

1100

15

460

-

A

-

A

-

A

-

A

-

A

-

A

-

A

12

23

240

3

93

-

A

-

A

-

A

-

A

-

A

-

A

-

A

 

 

For figures – click here

 

 

REFERENCES

 

Bartram, J. & S. Pedley (1996). Multiple fermentation tube technique 10.3. In: Bartram, J. & R. Balance (eds.). Water Quality Monitoring - A Practical Guide to the Design and Implementation of Freshwater Quality Studies and Monitoring Programmes.United Nations Environment Programme and the World Health Organization, 383pp.

Binzer, A., C. Guill, B.C. Rall & U. Brose (2016). Interactive effects of warming, eutrophication and size structure: impacts on biodiversity and food-web structure. Global Change Biology 22(1): 220–227.

Boles, G.L. (1981). Macroinvertebrate colonization of replacement substrate below a hypolimnial release reservoir. Hydrobiologia 78(2): 133–146. https://doi.org/10.1007/BF00007587

Botts, P. (1997). Spatial pattern, patch dynamics and successional change: chironomid assemblages in a lake Erie coastal wetland. Freshwater Biology 37(2): 277–286. https://doi.org/10.1046/j.1365-2427.1997.00145.x

Britton, L.J. & P.E. Greeson (1989). Methods for Collection and Analysis of Aquatic Biological and Microbiological Samples. Techniques of Water-Resources Investigations of the U.S. Geological Survey, Book 5. 172pp.

Carpenter, S.R., N.F. Caraco & D.L. Correll (1998). Nonpoint pollution of surface waters with phosphorus and nitrogen. Ecological Applications 8(3): 559–568.

Ceschin, S., V. Zuccarello & G. Caneva (2010). Role of macrophyte communities as bioindicators of water quality: application on the Tiber River basin (Italy). Plant Biosystems 144(3): 528–536. https://doi.org/10.1080/11263500903429221

Connolly, N.M., M.R. Crossland & R.G. Pearson (2004). Effect of low dissolved oxygen on survival, emergence, and drift of tropical stream macroinvertebrates. Journal of the North American Benthological Society 23(2): 251–270. https://doi.org/10.1899/0887-3593(2004)023<0251:EOLDOO>2.0.CO;2

DNPWC (2019). Langtang National Park. Department of National Parks and Wildlife Conservation Nepal. https://dnpwc.gov.np/protected_areas/details/langtangnationalpark. Accessed 22 April 2019.

ESRI (2014). ArcGIS Desktop: Release 10. Date of access is 2 November 2016.

Foote, A.L., S. Pandey & N.T. Krogman (1996). Processes of wetland loss in India. Environmental Conservation 23(1): 45–54. https://doi.org/10.1017/S0376892900038248

Fore, L.S., J.R. Karr & R.W. Wisseman (1996). Assessing Invertebrate Responses to Human Activities: Evaluating Alternative Approaches. Journal of the North American Benthological Society 15(2): 212–231.

Fox, J., P. Yonzon & N. Podger (1996). Mapping conflicts between biodiversity and human needs in Langtang National Park, Nepal. Conservation Biology 10(2): 562–569. https://doi.org/10.1046/j.1523-1739.1996.10020562.x

Gautam, B., R. Maskey, R.P. Sapkota & D.R. Dangol (2014). Aquatic macro-invertebrates as bio-indicators: an approach for wetland water quality assessment of Rampur Ghol, Chitwan, Nepal. Journal of Institute of Science and Technology 19(2): 58–64. https://doi.org/10.3126/jist.v19i2.13853

Gerlitz, J.Y., S. Banerjee, N. Brooks, K. Hunzai & M. Macchi (2015). An approach to measure vulnerability and adaptation to climate change in the Hindu Kush Himalayas, pp151–176. In: Filho W.L. (eds.). Handbook of Climate Change Adaptation. Springer, Berlin, Heidelberg, 2198pp. https://doi.org/10.1007/978-3-642-38670-1_99

Glime, J.M., R.G. Wetzel & B.J. Kennedy (1982). The effects of bryophytes on succession from alkaline marsh to sphagnum bog. American Midland Naturalist 108(2): 209–233. https://doi.org/10.2307/2425482

Google Earth (2016). Gatlang, Rasuwa, Nepal. Lat 28.156331, long 85.261294. DigitalGlobe, V. 7.3.2. Avaialble online at http://www.earth.google.com. Accessed on 1 November 2016.

Gotelli, N.J. & A. Chao (2013). Measuring and estimating species richness, species diversity, and biotic similarity from sampling data, pp195–211. In: Levin S.A. (ed.) Encyclopedia of Biodiversity, second edition, Volume 5. Academic Press, Waltham, MA.

Gutser, R., T. Ebertseder, A. Weber, M. Schraml & U. Schmidhalter (2005). Short-term and residual availability of nitrogen after long-term application of organic fertilizers on arable land. Journal of Plant Nutrition and Soil Science 168(4): 439–446. https://doi.org/10.1002/jpln.200520510

IPCC (2007). Contribution of Working Group II to the Fourth Assessment Report of the Intergovernmental Panel on Climate Change, 2007. In: Parry, M.L., O.F. Canziani, J.P. Palutikof, P.J. van der Linden & C.E. Hanson (eds.). Cambridge University Press, Cambridge, United Kingdom and New York, NY, USA. https://www.ipcc.ch/report/ar4/wg2/. Accessed 2 September 2017.

IUCN (2018). The IUCN Red List of Threatened Species, Version 2018-2. Avaiable online at www.iucnredlist.org. Downloaded on 1 April 2019.

IUCN (2016). The IUCN Red List of Threatened Species, Version 2016-3. Avaialble online at www.iucnredlist.org. Downloaded on 1 April 2019. 

Jansson, A. (1977). Distribution of Micronectae (Heteroptera, Corixidae) in lake Päijänne, central Finland: correlation with eutrophication and pollution. Annales Zoologici Fennici 14(2): 105–117.

Khan, M.A., M.A. Shah, S.S. Mir & S. Bashir (2004). The environmental status of a Kashmir Himalayan wetland game reserve: aquatic plant communities and eco-restoration measures. Lakes and Reservoirs: Research and Management 9(2): 125–132. https://doi.org/10.1111/j.1440-1770.2004.00242.x

Kharel, F.R. (1997). Agricultural crop and livestock depredation by wildlife in Langtang National Park, Nepal. Mountain Research and Development 17(2): 127. https://doi.org/10.2307/3673827

Li, X. & R. Guo (2017). Comparison of nitrogen removal in floating treatment wetlands constructed with Phragmites australis and Acorus calamus in a cold temperate zone. Water, Air & Soil Pollution 228(4): 132. https://doi.org/10.1007/s11270-017-3266-z

Malla, S.B., A.B. Shrestha, S.B. Rajbhandari, T.B. Shrestha & P.M. Adhikari (1976). Flora of Langtang and cross section vegetation survey (central zone). Kathmandu Department of Plant Resources. Dept. of Medicinal Plants, Thapathali, Kathmandu (Nepal) Ministry of Forests.

Manandhar, Y. (2007). Parbati Kunda, Rasuwa. World WildlifeFund. Available online at http://wwfnepal.org/?170420/Parvati-Kunda-Booklet#. Accessed on 01 October 2016.

Merrey, D.J., A. Hussain, D.D. Tamang, B. Thapa & A. Prakash (2018). Evolving high altitude livelihoods and climate change: a study from Rasuwa District, Nepal. Food Security 10(4): 1055–1071.

Ministry of Physical Planning and Works (2005). National Drinking Water Quality Standards and Directives.Nepal Department of Water Supply and Sewerage.Downloaded on 23 January 2018.  http://mowss.gov.np/assets/uploads/files/NDWQS_2005_Nepal.pdf

Murray, A.B. (2009). A Manual for an Inventory of Greater Himalayan Wetlands. International Centre for Integrated Mountain Development,Kathmandu, Nepal, 57pp.

Nurminen, L. (2003). Macrophyte species composition reflecting water quality changes in adjacent water bodies of lake Hiidenvesi, SW Finland. Annales Botanici Fennici 40: 188–208.

Pandit, A.K. (1999). Freshwater Ecosystems of the Himalaya. Parthenon, New York, 207pp.

Prasad, S.N., T.V. Ramachandra, N. Ahalya, T. Sengupta, A. Kumar, A.K. Tiwari, V.S. Vijayan & L. Vijayan (2002). Conservation of wetlands of India: a review. Tropical Ecology 43(1): 173–186.

R Development Core Team (2016). R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing. Available online at http://www.r-project.org.

Ralph, C.J., S. Droege & J.R. Sauer (1995). Managing and monitoring birds using point counts: standards and applications. In: Ralph, C.J., J.R. Sauer & S. Droege (technical editors). Monitoring bird populations by point counts. Gen. Tech. Rep. PSW-GTR-149. Albany, California: US Department of Agriculture, Forest Service, Pacific Southwest Research Station: p. 161–168, 149.

Rickey, M.A. & R.C. Anderson (2004). Effects of nitrogen addition on the invasive grass Phragmites Australis and a native competitor Spartina pectinata. Journal of Applied Ecology 41(5): 888–896. https://doi.org/10.1111/j.0021-8901.2004.00948.x

Rion, V., J.D. Gallandat, J.M. Gobat & P. Vittoz (2018). Recent changes in the plant composition of wetlands in the Jura Mountains. Applied Vegetation Science 21(1): 121–131. https://doi.org/10.1111/avsc.12338

Romshoo, S.A. & I. Rashid (2014). Assessing the impacts of changing land cover and climate on Hokersar Wetland in Indian Himalayas. Arabian Journal of Geosciences 7(1): 143–160. https://doi.org/10.1007/s12517-012-0761-9

Schindler, D.W. (2006). Recent advances in the understanding and mnagement of eutrophication. Limnology and Oceanography 51(1): 356–363.

Tachamo-Shah, R.D., D.N. Shah & H. Nesemann (2011). Development of a macroinvertebrate-based Nepal Lake Biotic Index (NLBI): an applied method for assessing the ecological quality of lakes and reservoirs in Nepal. International Journal of Hydrology Science and Technology 1(1–2): 125–146.

Tse-ring, K., E. Sharma, N. Chettri & A. Shrestha (2010). Climate Change Impact and Vulnerability in the eastern Himalayas: Synthesis Report. International Centre for Integrated Mountain Development (ICIMOD), Kathmandu, Nepal, 30pp.

Weber, M. & R. Brändle (1996). Some aspects of the extreme anoxia tolerance of the Sweet Flag, Acorus calamus L. Folia Geobotanica 31(1): 37–46. https://doi.org/10.1007/BF02803992

Xu, J., A. Shrestha, R. Vaidya & M. Eriksson (2007). The Melting Himalayas: Regional Challenges and Local Impacts of Climate Change on Mountain Ecosystems and Livelihoods. InternationalCentre for Integrated Mountain Development (ICIMOD), Kathmandu, Nepal, 13pp.

Xu, J., R.E. Grumbine, A. Shrestha, M. Eriksson, X. Yang, Y. Wang & A. Wilkes (2009). The melting Himalayas: cascading effects of climate change on water, biodiversity, and livelihoods. Conservation Biology 23(3): 520–530. https://doi.org/10.1111/j.1523-1739.2009.01237.x

Yao, T., Y. Wang, S. Liu, J. Pu, Y. Shen & A. Lu (2004). Recent glacial retreat in high Asia in China and its impact on water resource in northwest China. Science in China, Series D: Earth Sciences 47(12): 1065–1075. https://doi.org/10.1360/03yd0256  

Zhang, X., D. Wei, Z. Hu, A. Zhang, F. Li, J. Zhang, Y. Wu & Y. Tang (2018). Tolerance and physiological responses of Sweet Flag (Acorus calamus L.) under nitrite stress during wastewater treatment. Ecological Engineering 122: 107–111. https://doi.org/10.1016/j.ecoleng.2018.07.032