Biological Indicators, Habitat Classification and Its Assessment

Studies on Spring Conservation: Biological Indicators, Habitat Title Classification and its Assessment( Dissertation_全文 )

Author(s) Sun, Ye

Citation 京都大学

Issue Date 2020-03-23

URL https://doi.org/10.14989/doctor.k22610

Right

Type Thesis or Dissertation

Textversion ETD

Kyoto University

Studies on Spring Conservation: Biological Indicators, Habitat Classification and its Assessment

湧水保全に関する研究

ー生物指標種、生息地分類及びアセスメントー

（正確に記入すること。論文タイトルの和訳または英訳を付記しない。）

孫燁

【資料 3】申請者

【内表紙】

博士（総合学術）

Studies on Spring Conservation: Biological Indicators, Habitat Classification and its Assessment

湧水保全に関する研究ー生物指標種、生息地分類及びアセスメントー

孫燁

京都大学大学院総合生存学館

2020 年 3 月

CONTENTS

TABLE OF CONTENTS ...... 1

ABSTRACT ...... 3

INTRODUCTION ...... 5

i. Springs in the Landscape...... 5

(1) Biodiversity Values of Springs ...... 6

(2) Social and Cultural Values of Springs ...... 7

ii. Challenges to Springs ...... 8

(1) Spring Degradation and River Management ...... 8

(2) Methods for Groundwater and Spring Monitoring ...... 9

(3) Riverine Spring Classification ...... 10

iii. Objectives and Scope of Research ...... 12

CHAPTER 1

Development of Spring Indicator of Benthic Invertebrate Taxa ...... 13

1.1 A Broad-Scale Survey of Benthic Invertebrates in Springs ...... 13

(1) Data Collection of Spring Fauna ...... 13

1.2 Results of Investigating Invertebrate Fauna of Springs ...... 22

(1) Taxonomic Composition of Spring Fauna...... 22

(2) Spring Indicators of Benthic Invertebrates ...... 28

1.3 Applicatin of Spring Indicator ...... 32

1.4 Conclusion ...... 35

CHAPTER 2

Classification of Riverine Spring Habitats and Fauna Characteristics ...... 36

2.1 Spring Classification in this Study ...... 36

2.2 Study Sites and Data Collection ...... 39 1

(1) Spring-Flow Type: Hodakanomori, Gamada River ...... 40

(2) Floodplain Spring: Hiru Valley, Gamada River ...... 42

(3) Water’s Edge Spring: West Side of Kamogamo Shrine, Kamo River ...... 44

(4) Under-Water Spring: East Side of Shimogamo Shrine, Kamo River ...... 45

2.3 Data Analysis...... 46

2.4 Biological Differences Among Spring Habitat Types ...... 49

(1) Taxonomic Composition ...... 49

(2) The Patterns of Invertebrate Diversity ...... 51

(3) The Patterns of Ecological Types ...... 52

2.5 Discussion ...... 55

2.6 Conclusion ...... 59

CHAPTER 3

Application to Conservation of Spring Ecosystems and Environmental Education .... 60

3.1 Spring Monitoring ...... 61

3.2 Spring Habitat Assessment ...... 66

3.3 Environmental Education Project ...... 67

(1) Project Overview ...... 68

(2) Project Planning ...... 68

(3) Project Implementation ...... 70

(4) Project Accomplishments ...... 71

Conclusions and Future Directions...... 73

Acknowledgments...... 74

References ...... 75

Appendix 1...... 84

Appendix 2...... 121

ABSTRACT

Freshwater springs are a significant component of basin landscapes. They have an important role in sustaining biodiversity in aquatic systems, which intrinsically link to human welfare by providing various ecosystem services. However, spring habitats have been under risk of deterioration by land reclamation and water resource development. Therefore, the conservation of springs should be considered an important part of integrated basin management. In this thesis, we focus on the key elements to improving spring management including identification of biological indicators of spring fauna, classification of riverine spring habitats and identification of their ecological roles, and provision of methods for spring assessment in basin management.

1. The role of groundwater in the surface water ecosystem is not fully understood. Groundwater can be easily affected by land reclamation, water resource development, and climate change. The future policy is needed to better understand the interaction between groundwater and surface water. We present biological indicators of benthic invertebrates to evaluate the contribution of groundwater to surface water bodies. Because the benthic community is so dependent on its surroundings and strongly affected by its environment, including sediment composition and quality, water quality, as well as hydrological factors that influence the physical habitat, it serves as a biological indicator that reflects the overall condition of the aquatic environment.

We collected data of benthic invertebrates from both field research and literature at a continental and world-wide scale. By analyzing their taxonomic and ecological types, we identified spring indicator taxa of benthic invertebrates based on their dependent degree to the groundwater environment.

A total of 1,448 aquatic invertebrate species representing 58 orders were found from 249 research sites. The spring indicators were identified as spring dependent species including groundwater species (Stygobites and Stygophiles), cave species (Troglobites and Troglophile), and stenothermal species. Considering the geographical distribution patterns of the spring indicator taxa, stenothermal species were classified into "cold stenothermal species" which evolutionarily originate in more boreal regions and "warm stenothermal species" derived from more tropical regions. The ecological interpretation of these stenothermal species was discussed in relation to climatic zones and the altitude of the basin concerned. Based on the variations of spring contribution into river ecosystems, suggested by the spring indicator species, we proposed an application procedure of the spring indicators for environmental assessment and nature conservation works in river management.

2. Springs that are hydro-geologically connected to river channels are considered to have different spatial dimensions of interaction with surface water. Such interactions create a mosaic of inner- connected micro-habitats that play an important ecological role in structuring benthic invertebrate

3 assemblages. However, little is known about the spring typological variations and their ecological roles in a river system.

We have identified two major spring types within the braided river landscape, based on their locations in relation to the main river channel and flood plain. These are the following:

Spring-flow type: the spring emerges from outside of the one-year-floodplain zone, forming spring flows into a mainstream channel.

Riverbed spring type: the spring emerges within the riverbed (The riverbed is defined as areas within the one-year-floodplain zone).

Based on the relative relationship between spring location and the water level of the mainstream channel, the riverbed spring is classified into three types:

(1) Floodplain spring: spring emerges within the upper zone of the riverbed.

(2) Water’s edge spring: spring emerges on the water’s edge at low-water-level.

(3) Under-water spring: spring emerges under the water in the mainstream channel.

Field research and literature studies were conducted to identify biological values across different spring habitat types. Taxonomic composition, species richness, biodiversity patterns, and ecological types (matrix type, lifestyle, functional feeding group, and food type) of benthic invertebrates were analyzed. The results showed distinct biodiversity patterns and ecological types in spring habitats.

We found high levels of species richness in springs located outside of the floodplain zone and springs emerging underwater in the main river channel. Springs that emerge in the upper zone within the floodplain zone appeared to have higher biodiversity than those close to the river channel. In addition, the results demonstrated distinct patterns of ecological types associated with environmental conditions in spring habitat variations. Thus, it is suggested that spring conservation and river management should recognize the dynamic interactions between springs and surface water in riverine spring habitats and their important consequences for biodiversity and ecological types of invertebrates.

3. There are many factors and multi-stakeholders engaging in river management practice. Experts and researchers would best serve the spring conservation and decision-making process by providing information and suggestions to decision-makers and the public. We suggest that an effective spring assessment and sustainable basin management should consider the hydrogeological context of springs and their ecological values. We recommend biological indicator and regional spring typology as key steps of the basis of spring monitoring and assessment. Advice is given for researchers engaging environmental education to raise public environmental awareness, and a project report provides an extended example of the action research process.

INTRODUCTION

Groundwater emerges to the earth's surface, it becomes spring. As an important part of our landscape, springs play an important role in the health and longevity of our society as well as our planet. However, this integrating property also exposes springs a range of direct and indirect human activities (Scarbrook et al., 2007). River management and land use practice should carefully consider the values of springs and balance the conflicting uses when determining management actions to protects.

Section i provides the information on biological, social and cultural values of springs in our landscape. Section ii gives an introduction of challenges to springs using the example of the Kamo River to address the importance of sustainable spring management. Section iii and section v illustrate the purpose and the scope of this study.

i. Springs in the Landscape The special characteristics of springs, the locations of springs among groundwater ecosystems, surface water, and terrestrial ecosystems have led to a high value of biodiversity and a high contribution to ecosystem services (Figure 1). Despite the relatively small area of springs within the landscape, these ecosystems support a high number of endangered species and rare groundwater- dependent species. Springs serve as refugia for aquatic species from the river ecosystem. Springs also hold great cultural significance and have a great contribution to cultural services in terms of spiritual and historical experiences, scientific discovery and education.

Figure 1 The role of springs among ecosystems.

(1) Biodiversity Values of Springs

It is well known that the different physical, chemical and biological factors in the river ecosystem influence the distribution pattern of aquatic organisms. Springs and spring-fed streams are respectively regarded as habitats for some characteristic aquatic organisms in the river ecosystem.

Springs and spring-fed streams are stable in water temperature. They have a relatively long-term temperature of the aquifer. The spring water is known as cooler in summer and warmer in winter, comparing to surface water. Springs, spring-fed rivers and streams are important habitats for coldwater fish species. Coldwater fish species such as coho salmon and rainbow trout are increasingly threatened by climate change. As water temperature rises, they lost their habitats and must adapt or migrate to other habitats. However, freshwater fish cannot migrate too far and the river corridors, barriers and physical conditions (saltwater tolerance, temperature, etc.) also prevent them from migration. If they cannot adapt or migrate, the loss of habitats will eventually lead to their extinction. Springs, spring-fed rivers and streams are becoming more and more important for cold-water fish species because the water volume and temperatures in these river systems are more resilient to variation in precipitation and climate change than surface runoff watersheds. Springs and spring-fed streams are proving these species the cold-water refuges to survive as climate changes and the water temperature of surface water-fed streams becomes warm.

The environmental conditions in the springs and spring-fed streams are relatively stable. They are stable in water flow, sediment dynamics and disturbance through flood seasons and supra-seasonal drought (Stubbington, et al. 2013). Therefore, springs and spring-fed streams are acting as refuges for some aquatic organisms during the flood seasons and supra-seasonal drought seasons.

The content of spring water depends on the nature of the geology through which it passes. Springs and spring-fed streams usually contain minerals as they move through the underground rocks, minerals become dissolved in the water. Some spring water has significant amounts of minerals, and some spring water contains significant amounts of dissolved sodium salts. Besides the physical and chemical stability, springs and spring-fed streams are also shown to have smaller and more isolated habitat areas, and fewer large predators, compared to higher-order streams (Glazier, 1991). Because of these physical and chemical characteristics, springs and spring-fed streams become unique habitats (refuges) in the realm of running waters. Springs and spring-fed streams are found to be characterized by distinctive aquatic species communities and marked heterogeneity of environmental conditions and communities (Gray, et al. 2011). Several studies showed that springs sustained high levels of biodiversity of aquatic organisms (e.g., Illies, 1978). Some previous studies also reported that spring water is acting as hot spots of aquatic biodiversity and productivity in the river ecosystem, because springs are connections of the groundwater system and interface water system (Cantonati et al., 2006; Scarsbrook et al., 2007).

(2) Social and Cultural Values of Springs

Springwater has strongly linked with human life as an indispensable environmental resource for basic needs, accumulating historical values of the relationship between water and human. Especially cool springs have devoted to human life and culture. Therefore, springs have historical values for communities across the globe and the potential values can go beyond our imagination that requires our constant attention to preserve them.

It is easy to consider that clean water had played a far more critical role in human life and played a central role in survivability before the accomplishment of a modern water system. For instance, it is said that some aboriginal people in Australia have built travel routes between different springs and these springs have promoted their trade for their daily use such as stones to make weapons and tools, foods or so on. Many of the sites still have been utilized as community gatherings and ceremonies, such as initiation rituals, funerals, and marriages1.

Also, in Japan location of springs influenced architectures of ancient castles and systems of towns since freshwater resources were used by people like drinking, cooking, and washing. Shinokura Shrine is one of the examples which tells the belief of citizens on spring water as an important material for curing eye disease. Thus, Japanese shrines have various stories that are closely tied with rituals, festivals, and historical monuments. In the Wakasa area, according to folklore, an Indian monk named Jicchu built Todayji Nigatsu-do and invited deities in celebrating the construction of the great Buddha. One of the deities was late for the celebration and he swore to give spring water in compensation. The well was later called 'Wakasai’, is a symbolical value for the community even today.

Jeju Island in Korea, spring water was a daily need of the people until modern water facilities were introduced in the 1980s. Domestic and agricultural water was provided by the spring. Thereby, most villages were formed along with the location where spring water was abundant2. Certain folk beliefs found on Jeju Island states that the water was used for nursing mothers when their breast milk was not sufficient. Bile is a word in the Jeju Island dialect and means a flat and wide rock on the ground. Water springing out from rock is called Gomang3-mul, Goet-mul or Bile-mul in the local language and Jeol-mul indicates spring water near the pagoda.

The aforementioned examples denote how spring water is strongly related to human history and culture, showing its significant value worth to protect from the view of cultural service.

1 Cultural Values of the Great Artesian Basin Fact Sheet, Great Artesian Basin Coordinating Committee, Australia, 2016. URL: www.gabcc.gov.au/publications/cultural-values-fact-sheet. 2 Park, W. B., Ha, K., 2012. Springwater and water culture on Jeju Island. Groundwater 50 (1), 159-165. 3 Gomang is a word in the Jeju Island dialect that indicates a hole. 7 ii. Challenges to Springs

(1) Spring Degradation and River Management

The special location of springs among the groundwater system and surface water system leads to many challenges in spring management. River management and regulation are considered as important factors that can influence the health of springs. Diversion, channelization, and impoundment can give severe impacts on braided river springs (Scarsbrook et al., 2007). Until recently, very little research has focused on springs ecosystems or their dependent species. This lack of information and attention to springs ecosystems has resulted in the loss of many springs through poor groundwater and land-use practices.

In Kamo River (Kyoto, Japan), instream springs and small spring-fed ponds were serving as habitats and refugia for coldwater species, such as Plecoglossus altivelis (Picture 1). After the river channel has been dredged for flood control, sediments flowed along from the up-streams have been dredged. Some small ponds of braided river springs surrounded by sediments lost their protection and mixed with surface water. The loss of such spring habitats caused much death of fishes and creatures that rely on those spring habitats (Picture 2). Losing springs habitat can cause diversity loss and environmental crisis. The conservation of springs is essential both for the value of biodiversity and their functional roles.

Picture 1 Spring pond surrounded by sediments in Kamo River and species (Plecoglossus altivelis) that uses springs as refugia in summer seasons (Photo by Nakasuji Y., 2011).

Picture 2 Died fishes led by the spring habitat loss (Photo by Fujibayashi, K., 2017).

(2) Methods for Groundwater and Spring Monitoring

Springs are completely fed by groundwater. The permanence of springs relies on the constant water supply of groundwater. Streams, lakes, and wetlands are discharged by groundwater continuously or occasionally in terms of springs. Therefore, monitoring groundwater flow, evaluating the contribution of groundwater to surface water system is essential for spring conservation and river basin management.

Models, Simulations and Direct Field Measurements

With the increasing attention for the importance of interaction between groundwater and surface water bodies, several direct and indirect methods have been conducted to evaluate the contribution of groundwater to surface water systems in previous studies. Rozemeijer et. al. (2010) used direct filed- scale measurements of groundwater flow route contribution to surface water contamination. The groundwater discharge was separately captured from the tube drain effluent in the filed by a novel experimental setup. Sherlyn (2004) estimated groundwater discharge to streams using indirect methods of hydrograph-separation techniques, drought-streamflow measurements, and linear- regression analysis of streamflow duration. The groundwater discharge data were then used in a groundwater model to evaluate groundwater flow. Heat tracer methods based on vertical and streamed temperature profiles and regional mass balancing approach based on measurements are also used as indirect methods to estimate the contribution of groundwater discharge to streams (Kalbus et al., 2006; Eertwegh et al., 2006).

Among these methods, indirect measurements combing simulations and models were widely used in previous researches to estimate the groundwater flow. The limitation is that most of these indirect methods required some certain assumptions thus it is rather difficult to estimate the discharge of groundwater on a local scale. For instance, the indirect method of hydrograph-separation assumes that the tracer concentrations of the individual flow routes are constant in time, while as a matter of fact, several studies have shown variable solute concentrations (Rozemeijer et al., 2009; Tiemeyer et al.,

2006; Langlois and Mehuys, 2003). Direct filed-scale measurements of groundwater flow play an important role in evaluating groundwater contribution to streams, however, it is difficult for some field researches which are operated under limited conditions within a limited time, to capture sufficient data for estimation.

Biological Indicator of Benthic Invertebrates

Benthic invertebrates include surface water species, groundwater-dependent species, and some wetland and terrestrial species that benefit from the groundwater environment inhabit in spring habitats (Danielopol and Pospisil, 2001). Because the benthic community is so dependent on its surroundings, strongly affected by their environment, including sediment composition and quality, water quality, and hydrological factors that influence the physical habitat, it serves as a biological indicator that reflects the overall condition of the aquatic environment. For example, benthic invertebrates have been used as indicator species to predict species richness of multiple taxonomic groups in previous studies (Fleishman et al., 2005). However, documentation of the above species and the methods of using them as biological indicators for groundwater and springs is scant.

(3) Riverine Spring Classification

Springs located among groundwater ecosystems, river ecosystems, and terrestrial ecosystems, are known as an important part of our landscape. They are characterized by marked heterogeneity of environmental conditions and distinctive aquatic species communities (Staudacher and Fu¨reder, 2007; Kubíková et al., 2012). Previous studies reported springs as hotspots of aquatic biodiversity, providing habitats for a myriad of aquatic organisms including surface water species, groundwater species and stenothermal species (Roca, 1993; Cantonati et al., 2006; Scarsbrook et al., 2007; Lencioni, 2008). As freshwater organisms evolve, adapting to the hierarchical habitat structure, it is essential to improve our understanding of the structure and function of different types of springs. Such classifications would lay the groundwork for regional conservation efforts (Springer et al., 2008).

The hydrogeological context has been seen as the determinant of spring structure and function, which drives the varied physicochemical characteristics of spring flows (van der Kamp, 1995). Previous studies have classified springs based on their geological characteristics (e.g., emergence environment, the sphere of discharge and channel dynamics) and physicochemical parameters (e.g., water temperature, discharge and groundwater flow paths). The classification of Byran (1919) was divided into classes of springs resulting from non-gravitational forces and springs resulting from gravitational forces, using the geologic structure and the origin of the water as the main criteria. Meinzer (1923) characterized springs by their discharge. Clarke (1924) proposed a classification of springs based on the criteria of geologic origin, physical properties, and chemical properties. Stiny (1933) described three main classes of springs based on flow. Fetter (1980) focused on the endpoint of 10 the groundwater flow path and divided springs into five main classes: depression spring, contact spring, fault spring, sinkhole spring and fracture spring. Alfaro and Wallace (1994), Wallace and Alfaro (2001); Springer et al. (2008); and Springer and Stevens (2009) reviewed and updated documentation and classifications of springs in use.

Barquin and Scarsbrook (2008) pointed out that the interaction between groundwater and surface water influences the structure and function of spring habitats. Indeed, the location of springs at the interface between groundwater and surface water drives a variety of physicochemical characteristics of spring habitats. As Kamp (1995) pointed out in his study, the interaction between groundwater and surface water may influence flow stability, thermal constancy and water chemistry, which may form the ecological roles of spring habitats in the context of a riverine system.

Riverine springs which are hydro-geologically connected to the river channels are considered to have different spatial dimensions of interaction: interactions from spring to river and interactions between spring and flood plain. Previous studies have shown such interactions play an important role in determining biodiversity patterns of benthic assemblages (Arscott et al., 2005; Malard et al., 2006), although at which point this occurs remains to be studied. Some studies showed that floods reduce invertebrate abundances and diversity (Burgherr et al., 2002; Reckendorfer et al., 2006). On the other hand, Gray et al. (2006) investigated braided channels, springs and hillslope streams in the Waimakariri River and the results showed that spring systems embedded within the floodplain zone have respectively high invertebrate biodiversity. Likens (2009) claimed that the species diversity of invertebrates increases from groundwater to surface spring habitats to downstream sites.

Although channel dynamic and flood events were pointed out as important factors that influence spring structure and function, little is known about the riverine spring typological variations and their ecological functions. Generally, springs flowing into one or more stream channels were defined as Rheocrene or flowing spring (Bornhauser, 1913; Hynes, 1970). Springer et al. (2008) illustrated the importance of continuum between spring and stream channels and classified Rheocrene into three types based on spring and runoff channel dynamics and morphologies: spring-dominated type which flows into the headwater of a stream where there is little runoff flow, runoff-dominated type which flows into a stream channel and has significant runoff contribution, and the intermediate type. The spring-dominated type tends to be relatively stable. The runoff-dominated type is considered to have more influences from flood events.

Rivers typically have flood events that occur at different points in time and create a dynamic flood plain zone. Such flood plain elements make spatially complex and temporally variable groundwater- surface water exchanges (Stanford and Ward, 1993; Brunke and Gonser, 1997; Poole et al., 2002). Floodwaters can carry surface water and eroded sediments of soils, which can influence the structure and functions of spring habitats. If a spring is located close to the floodplain zone, its water quality,

11 soil condition, vegetation types, density, and fauna features may be influenced by the floods. Consequently, these interactions create a mosaic of heterogeneous, connected micro-habitats that play an important role in shaping spatial patterns of benthic invertebrate biodiversity (Ward et al., 2002). Flood effect has been considered as an important factor in springs but few spring classifications take into account flood dynamics in spring classification. How, and to what extent the spring typology and ecology are related to flood elements remain largely untested.

iii. Objectives and Scope of Research

This thesis aims to examine the key elements of spring research and conservation including biological indicators, habitat classification and spring assessment c in different scales. By understanding the hydrogeological context and the ecological roles of springs in ecosystems, we aim to provide sufficient information and effective methods for sustainable spring management.

We conducted a broad-scale survey of benthic invertebrates in springs (Chapter 1). The data of benthic invertebrates were collected from both field research and literature at a continental and world- wide scale. By analyzing their taxonomic and their ecological types, the purpose was to propose a biological indicator list for monitoring groundwater and springs.

For the method of spring habitat assessment, we proposed a new classification of spring types, focusing on the locations of springs in relation to the main channel of rivers and their floodplain zone (Chapter 2). The location of springs within the floodplain of a river system drives the interaction between groundwater and surface water. The classification put forward in this paper stresses the different types of springs and their functions as habitats for organisms. The principal aim is to identify the characteristic of spring habitat types, highlight their ecological functions in the river ecosystem. In Chapter 3, we discuss the approaches on spring monitoring and spring assessment on a local spring- scale and a basin scale. We suggest that more attention should be paid on the application of biological indicators, local habitat classification and integrated basin management which contribute to spring research and conservation strategies.

CHAPTER 1 Development of Spring Indicator of Benthic Invertebrate Taxa

In this chapter, we focus on the development of biological indicators for spring management and conservation. Springs are suitable habitats for many characterized aquatic invertebrates which have closely dependent on their surrounding environments. The application of spring indicator taxa of benthic invertebrates provides information of an aquifer, which reflect the connections between surface water ecosystem and groundwater ecosystem.

Through a broad-scale of literature review of spring information and their benthic fauna, we collect benthic invertebrate data from previous research and field research at 249 sampling sites. We analyze the fauna composition and identify spring indicator taxa according to their relations to groundwater and spring water. Section 1.2 shows the results of investigating spring indicator taxa. Further discussion is held to classify the application possibility of spring indicator of benthic invertebrate taxa in biodiversity conservation and sustainable spring management.

1.1 A Broad-Scale Survey of Benthic Invertebrates in Springs

(1) Data Collection of Spring Fauna

Macroinvertebrate data were extracted from the most recent publications regarding macroinvertebrate (benthic invertebrate) and spring habitats and two original field surveys conducted in Japan. The data list was created also with the aid of the Web of Science Core Collection (TR) database and the recent checklist of the recent freshwater ostracod fauna (Karanovic, 2012). We selected 249 sampling sites mainly from two water body types: springs and spring-fed flows (Table 1). Throughout this paper, we refer to springs as sampling sites located within a few meters of the groundwater discharge sources. The spring-fed flows refer to the sampling that occurred in the Springbrook or flows discharged by groundwater. The reference list also contains sampling sites of the river. Since some research papers also contain sampling sites of rivers, we selected those data as supplementary references.

The sampling sites were classified into three climate zones: tropical zone, temperate zone and boreal zone, based on the thermal conditions in the study area. According to the theory of Aristotle, the tropical zone in this paper is defined as the region from 23.5ºN to 23.5ºS, the temperate zone is from 23.5ºN to 66.5ºN and 23.5ºS to 66.5ºS. Aristotle dubbed the frigid zone as the area north of the Arctic Circle (66.5ºN) and south of the Antarctic Circle (66.5ºS), as he reasoned that these regions

13 were permanently frozen and uninhabited. The north of the Arctic region and south of the Antarctic region are indeed inhabitable for most aquatic benthic invertebrates. Considering the limited data set of invertebrates could be possibly collected from these area, we decided to divide invertebrate data of area north of 60ºN and south of 60ºS into the frigid zone.

In total, 1448 taxa were identified from the 249 sampling sites. We classified data of benthic invertebrates using phylogenetic methods. The taxonomic rank of phylum, class, order, family, genus and species are identified in each invertebrate organism. For each taxonomic unit, there is a diagnosis, provided based on the most recent publications dealing with the taxon and the World Register of Marine Species (http://www.marinespecies.org).

Table 1 Locations and water body types of reviewed paper in this study (water body types: S=spring, F=Spring-fed flow, R=river).

Number of Tropical Temerate Frugud sites Country River site References zone zone zone S F R No.

Asia Japan ◎ Tsuya stream, Gifu 1 1 Abdelsalam and Tanida (2013) 1 Japan ◎ Tama River basin, Tokyo 44 Shinoda (2007) 2 Japan ◎ Gamata River, Gifu 1 Nomura and Takemon (2007) 3 Japan ◎ Gamata River, Gifu 4 Sun et al. (2018) 4 Japan ◎ Kamogawa River, Kyoto 4 Suzuki (2007) 5 Japan ◎ Kakita River, Shizuoka 1 Takemon (2010) 6 China ◎ Jiuzhaigo Nature Preserve, Yunnan 1 Zhang et al. (2005) 7

Yunan, Sichuan, Guizhou, Guangxi, Hunan, 8 China ◎ Li et al. (2007) 8 Hubei, Anhui, Zhejiang

15 Krakow-Cz ̨estochowa Upland, southern 23 Europe Poland ◎ Dumnicka (2007) 9 Poland Krakow-Cz ̨estochowa Upland, southern 3 6 Poland ◎ Dumnicka (2013) 10 Poland Poland ◎ Lubuska Upland, central-western Poland 21 RYCHŁA (2015) 11 Slovenia ◎ Sava River, Soca River (Julian Alps) 2 Mori and Brancelj (2006) 12 Austria ◎ Konigsbach stream, Tyrolean Alps 1 Füreder et. al. (2001) 13 Austria ◎ Landslide area"Schutt", Carinthia 1 Staudacher and Füreder (2007) 14 Finland ◎ Kiikalannumi groundwater area 5 Ilmonen and Paasivirta (2005) 15 United 2 1 ◎ River Wye catchment, Peak Distinct Smith and Wood (2002) 16 Kingdom United 5 ◎ River Wye catchment, Peak Distinct Smith et al. (2003) 17 Kingdom United 2 2 ◎ Little Stour River, Kent Stubbington and Wood (2013) 18 Kingdom United 1 ◎ Peak District Stubbington et al. (2009) 19 Kingdom

Table 1 (continued). Number of Tropical Temerate Frugud sites Country River site References zone zone zone S F R No.

Europe Spain ◎ Saja and Ason Natural Parks, Cantabria 6 Barquín and Death (2009) 20 Spain ◎ Saja and Ason Natural Parks, Cantabria 1 Barquín and Death (2004) 21 Sweden ◎ Glaciofluvial, Moraine, Limestone Springs 3 Hoffsten and Malmquvist (2000) 22 Switzerland ◎ Swiss National Park 10 20 Fumetti and Blattner (2017) 23 Switzerland ◎ Springs located around the city of Basel 1 Fumetti et al. (2007) 24 Republic of 1 ◎ Gacka River, karstic springs Matić et al. (2016) 25 Croatia 4 Springs in Parma River catchment (Cirone, 4 Italy ◎ Bottazzi et al. (2011) 26

Lagdei, Vezzosa, Biam) Germany ◎ Pfalzerwald mountains 1 Hahn (2000) 27

16 Luxembourg ◎ Spring 1 Martin and Stur (2006) 28

France ◎ Mercantour National Park, Alpes-Maritimes 6 Dole-Olivier et. al. (2015) 29 France ◎ Mercantour National Park 1 Martin et al. (2015.) 30 Turkey ◎ Lake District 1 Yıldız and Balık (2005) 31 Denmark ◎ River Gjern, headwater 1 Iversen et al. (1991) 32 Serbia ◎ Spring Pavkovac,Lezimir,Fruska Gora 1

Ivory coast ◎ Springs 1 Karanovic (2012), Martens and 33 Savatenalinton (2011) Spring on the hill above the village, Gornja 1 Montenegro ◎ Seoca Russia ◎ Springs in the south of Irkutsk area 1 Takhteeva et al. (2010) 34 Mineral Springs in the Kirenga River Basin 1 Russia ◎ Takhteev et al. (2017) 35 and the Upper Reaches of the Lena River Russia ◎ The headwaters of the Volga, boreal zone 1 Schletterer et al. (2014) 36

Table 1 (continued).

Number of Tropical Temerate Frugud sites Country River site References zone zone zone S F R No. 2 1 North America United States ◎ Comal, Hueco and Fern Band Springs, Texas Gibson et al. (2008) 37 United States ◎ Spring habitats, Bridge Creek 1 1 Anderson and Anderson (1995) 38 1 United States ◎ John Bryan State Park, Greene County, Ohio Butler M.J. and Hobbs H.H. (1982) 39 United States ◎ Putnam County, Tennessee 1 Stern and Stern (1969) 40 United States ◎ Cone Spring, Iowa 1 Tilly (1968) 41 United States ◎ Atomic Energy Reservation, Tennessee 1 Wilhm (1970) 42

United States ◎ San Marcos River, Texas 1 Perkin et al. (2012) 43 5 17 United States ◎ Huntingdon County, Pennsylvania Sangiorgi et al. (2010) 44

Ozark uplift region of Greene County, 3 United States ◎ Teresa et al. (2014) 45 Missouri, United States ◎ Headwaters 1 Grubbs (2011) 46 United States ◎ Black Oak Park stream, Ohio 1 McNeish et al. (2017) 47 Canada ◎ Oak Ridges Moraine, Ontario 1 Gathmann and Williams (2006) 48 Canada ◎ Valley Spring, Ontario 1 Williams and Hogg. (1988) 49 Canada ◎ Prince Edward Island 1 Dobrin and Giberson (2003) 50 Lander Springbrook, Roswell Artesian 1 South America Mexico ◎ Noel (1954) 51 Basin Fr ́ıo Basin in the mountains of the 1 Argentina ◎ Northwest of the Chubut Province in Brand and Miserendino (2011) 52 Patagonia

Table 1 (continued).

Number of Tropical Temerate Frugud sites Country River site References zone zone zone S F R No.

9 Springs(Ohinepango, Waitaiki, 1 Oceanic New Zealand ◎ Waihohonu,Taungatara, Slip, Hawdon, Cass, Barquín and Death (2011) 53 Pearse, Riwaka Resurgences),Nelson 2 1 New Zealand ◎ Slip spring, Cora Lynn Sream Death and Winterbourn (1995) 54

7 Springs(Waimak, One Tree Swamp, 7 New Zealand ◎ Hawdon Valley, O'malleys flat, Cora Lynn, Gray (2005) 55 Turkey Fan, Klondyke) Waikuku, Kaniwhaniwha, Gardeners Gut, 5

New Zealand ◎ Collier and Smith (2006) 56 Waitomo springs Bogong High Plains in the highlands of 1

1 Australia ◎ Clements et al. (2016) 57

8 north-eastern Victoria

Baharini Springbrook and Njoro River, 1 1 Africa Kenya ◎ Shivoga (2001) 58 Nakuru dolomitic springs in the North West Province 1 Karanovic (2012), Martens and Sourth Africa ◎ 33 (former western Transvaal) Savatenalinton (2011) Other World scale - Overijssel (EKOO) database 1 Bae et al. (2013) 59 Total 193 43 13

(2) Analysis of Fauna Composition and Identification of Spring Indicator Taxa

The fauna composition was analyzed to the level of species according to the phylogenetic classification. The basic units of phylogenetic classification are identified based on the physical characteristics of organisms and their genetics (`Winsor, 2009). The taxonomic rank of Phylum, Class, Order, Family, Genus and Species are identified in each invertebrate organism.

Spring indicators were identified as spring dependent species. We classified spring dependent species into three groups: groundwater-dependent species, cold stenothermal species, and other species that could be used as indicators.

Picture 3 Two groundwater dwellers (Eocrangonyx sp.) collected from the spring of the Kamo River, Kyoto (Photo by Takemon Y.)

Groundwater dependent species include a high number of diverse stygobites and stygophiles. Stygobites adapt to inhabit the subsurface environment and spend their whole life cycle in groundwater (Giber et al., 1994). They have developed life strategies to adapt to the subsurface environment in which light and resources are scarce. Their adaptation has resulted in the evolution of reduction or loss of eyes (blindness). They are also known as colorless or translucent

and having an elongation of body shapes to move among the voids within rocks (Picture 3). Stygophiles are organisms occur in both groundwater and surface water, while those that occur accidentally in groundwaters are identified as stygoxenes which are not considered as spring indicators in this study. Stygobites are strictly subterranean, and generally do not exist in surface waters. Stygophiles are not necessarily restricted to the subterranean environment, but they can exploit resources in groundwater (Maurice and Bloomfield, 2012).

Although there is no clear information on the geological and hydrogeological controls on groundwater fauna distributions, there are high numbers of groundwater species are known as limited-range endemics with low dispersal due to the limited underground conditions (Maurice and Bloomfield, 2012). Among the classification of functional feeding groups, decomposers and predators are significantly represented in groundwater fauna (Mohr and Poulson, 1996). The groundwater fauna can be sampled from different types of water bodies such as springs, spring- fed streams, hyporheic zone, caves and karst regions. The presence of groundwater fauna in surface waters could potentially be used as spring indicators.

Cold stenothermal species refers to cold-adapted stenotherms of benthic invertebrates, which grow over a narrow range of low temperatures, respectively. Cold stenotherms such as high polar

species have narrow thermal windows and low energy demand lifestyles (Field, 2014). They are commonly found in streams and rivers of high latitude regions, coldwater habitats of low latitude regions, such as headwater, springs, spring-fed and glacier-fed streams. The presence of cold stenothems in the temperate zone could potentially be used as spring indicators. Conversely, warm stenothermal species have adapted to a range of respectively high temperatures and can be commonly found in low latitude regions. Some stenothermal species of high latitude regions can also be considered as warm stenotherms. Because there are these special groups that could not survive if exposed to very freezing temperatures or sub-zero environments during the winter period. When the surrounding water freezes, the water temperature may go beyond their lethal limits of cold hardiness tolerance. In this case, they may find spring water as suitable habitats. The presence of such stenothermal species in high altitude regions during winter seasons could also potentially be used as local spring indicators. Species were classified as stenotherms if they were either known to occur naturally in cold streams, or high latitude regions. Studies on thermal tolerances of taxonomic groups of benthic invertebrates and their lifestyles were used to help identify the group of cold stenotherms.

Picture 4 Four coldwater dwellers (Metriocnemus sp.) found in the instream spring of Kamo River,

Kyoto (Photo by Takemon Y.).

Other species such as epiphytic species are also considered as possible spring indicators as they appear to inhabit spring water. Given the stability of springs as habitats, stones with moss mats are more common at spring sites. This could be one of the possible explanations for the abundance of stone surfaces and moss mat dwellers at springs and spring-fed streams. Many studies have also shown that the epiphytic species prefer cold waters. Tada and Satake (1994) reported epiphytic zoobenthos at the upper reaches of a cool mountain stream. Species Micrasema sp. MC was the most abundant taxon found on bryophyte mats at the headwater sites. Storey and Quinn (2017) have found the epiphytic species, Kempynus (Order Neuroptera) in small perennial headwaters. Within this context, epiphytic species share some common characteristics with stenothermal species.

Besides the above geographic, physical, and ecological factors that may explain why these groups can be considered as spring indicators, there is a group in which species have been recorded as possible spring indicators in the previous works, but the reasons maintained are unknown. During the biological sampling of benthic invertebrates in rivers and streams, some species which can be commonly recorded in spring waters are considered as spring species. However, the reasons why they inhabit springs are not clear and it could be geographic, 21

chemical, physical, ecological or other possible factors. This special group is also considered a possible spring indicator group in this study. The identification of this group refers to the current researches and documents.

1.2 Results of Investigating Invertebrate Fauna of Springs

(1) Taxonomic Composition of Spring Fauna

A total of 1448 aquatic invertebrate taxa representing 58 orders were found from 249 research sites. We identified 52 orders of benthic invertebrate taxa from research sites located in the temperate zone, 15 orders of fauna taxa from the tropical zone and 14 orders from the Frigid Zone. The fauna data in the temperate zone has Diptera, Trichoptera, Ephemeroptera, Plecoptera, Coleoptera, Amphipoda, and Hygrophila as the abundant Order (Figure 2). In the tropical zone, we collected research data mainly composed of Order Diptera, Hemiptera, Coleoptera, Ephemeroptera, Trichoptera, Odonata, and Diplostraca. In the Frigid Zone, Order Diptera, Coleoptera, Trichoptera, Plecoptera, Ephemeroptera, Odonata, and Hemiptera are found to be the common taxa.

The invertebrate faunas in spring habitats are dominated by Diptera and Trichoptera, and Plecoptera. Coleoptera, Ephemeroptera, Amphipoda and Isopoda are also relatively common (Figure 3). In Diptera, Chironomidae had the most number of species that occurred in springs, followed by

Limoniidae, Tipulidae, Dixidae, Simuliidae, Ceratopogonidae and Psychodidae. In Trichoptera, Limnephilidae had the most number of species that occurred in springs, followed by Hydrobiosidae, Conoesucidae, Hydroptilidae, and Polycentropodidae (Figure 4).

In spring-fed flows, invertebrate fauna was dominated by Diptera, Trichoptera Plecoptera and Ephemeroptera. In Diptera, Chironomidae was the most abundant Order, followed by Ceratopogonidae, Tanypodinae, Tipulidae, Diamesinae and Atyidae. In Trichoptera, Limnephilidae, Rhyacophilidae, Brachycentridae, Polycentropodidae, Psychomyiidae, followed by Hydropsychidae, Lepidostomatidae, Arctopsychidae, Conoesucidae, Glossosomatidae, Limnephiloidea, Philopotamidae, Goeridae, Helicophidae, Hydrobiosidae, Hydroptilidae, Leptoceridae and and Sericostomatidae. In Coleoptera, the most abundant families are Nemouridae, Chloroperlidae, and Perlidae (Figure 5).

Figure 2 Taxonomic composition of invertebrate fauna in Tropical Zone, Temperate Zone and Frigid Zone.

In rivers, the invertebrate fauna is mainly dominated by Diptera, Trichoptera, Ephemeroptera, Coleoptera and Plecoptera. In Diptera, Chironomidae was the most abundant family,

followed by Simuliidae, Tipulidae, Ceratopogonidae, Psychodidae, Empididae and Diamesinae (Figure 6). In Trichoptera, Limncephilidae was the most abundant family, followed by Leptoceridae, Hydropsychidae, Hydroptilidae, Glossosomatidae, and Goeridae. In Ephemeroptera, the most abundant families are Baetidae followed by Canada, Heptageniidae, Ephemerellidae, Ephemeridae, and Leptophlebiidae.

Order Diptera, Trichoptera, Plecoptera, Ephemeroptera and Coleoptera were found in three water bodies. The percentage of Order Plecoptera in spring and spring-fed habitats were higher than the river sites. In addition, compared to the taxa composition of river sites where Order Tubificida and Haplotaxida were found, spring habitats and spring-fed habitats were found to have a higher percentage of characterized invertebrate assemblage such as Order Amphipoda, Isopoda and Hygrophila. In the taxa composition of Order Diptera, Family Thaumaleidae and Sciomyzidae were characterized taxa in spring and spring-fed habitats. Several taxa were only found in the spring habitats, such as Pediciidae, Tabanidae, Sciaridae and Syrphidae. In the taxa composition of Order Trichoptera, several characterized taxa were only found in the spring habitats, such as Beraeidae, Oeconesidae, Molannidae, Philorheithridae, and Melanotrichie.

Figure 3 Taxonomic composition of invertebrate fauna collected from the three water bodies: A) Spring habitats, B) Spring-fed habitats, and C) Rivers.

Figure 4 Rank of families in Diptera, Plecoptera and Trichoptera by the number of occurrences from spring habitats.

Figure 5 Rank of families in Diptera, Plecoptera and Trichoptera by the number of occurrences from spring-fed habitats.

Figure 6 Rank of families in Diptera, Trichoptera and Ephemeroptera by the number of occurrences from spring-fed rivers

(2) Spring Indicators of Benthic Invertebrates

A total of 364 benthic invertebrate taxa representing 12 classes, 26 orders and 65 families are identified as spring dependent species (Appendix 1). Groundwater dependent species come from several different taxonomic groups from classes Copepoda, Malacostraca, Ostracoda, and Gastropoda. Within the Copepoda class, there are species of Harpacticoida (Canthocamptidae, Parastenocarididae) and Cyclopida (Cyclopidae) (Figure 7). Within the Malacostraca class, there are species from Amphipoda (Crangonyctidae, Niphargidae, Paraleptamphopidae, Pseudocrangonyctidae, Pontogeneiidae). Within the Ostracoda class, there are species from Podocopida (Candonidae, Cyprididae, Loxoconchidae). Within the Gastropoda class, there are species from Littorinimorpha (Hydrobiidae) and Heterostropha (Valvatidae).Canthocamptidae, Cyclopidae and Niphargidae are the most representative family.

Cold stenothermal species mainly come from class Insecta. Within the Insecta class, there are species of Diptera, Plecoptera (Capniidae, Chloroperlidae, Leuctridae, Nemouridae, Notonemouridae, Perlodidae, Perlidae, Gripopterygidae, Eustheniidae), and Coleoptera (Dytiscidae, Hydrophilidae). Other taxonomic groups include class Rhabditophora (Tricladida), and Malacostraca (Amphipoda). The Amphipoda order includes species from Gammaridae, Seborgiidae, Anisogammaridae, Hadziidae, Hyalellidae, and Mesogammaridae.Dytiscidae, Chironomidae and Nemouridae are the most abundant family. Order Trichoptera (Brachycentridae, Hydroptilidae) and Neuroptera (Osmylidae) are recorded

within epiphytic species.

Figure 7 Taxonomic composition of spring indicator taxa of benthic invertebrate.

Tricladida

There are currently 16 species in 2 families recorded within the Turbelaria (Tricladida) from present data collection of literature and field research. Among them, species in the genera Crenobia, Dugesia, Polycelis in Planariidae, Neppia in Dugesiidae are considered as cold stenothermal organisms. They appear to preferentially inhibit cold waters. They can be found both in headwater and springs. Cold stenothermal organisms may be spring indicators when they are observed in places that have a relatively high-water temperature (e.g., down streams). The remaining species recorded in Tricladida are considered stream dwellers that can be commonly found in surface waters and have no significant revelations with springs.

Gastropoda

A total of 120 families representing 10 orders within Gastropoda are reviewed. A total of 10 species in genus Bythinella, Graziana, Potamopyrgus, Belgrandiella, Bythinella and Iglica within Hydrobiidae (Littorinimorpha) are considered as spring indicators. The family Hydrobiidae referred to as spring snails are considered as spring specialists (Scarsbrook et.al., 2007). They can be found in springs and groundwater habitats (including cave resurgences and seepages).

The remaining families within 9 orders, Basomatophor (Lymnaeidae), Cycloneritmorp (Neritidae), Heterostropha (Valvatidae), Hygrophila (Lymnaeidae, Planorbidae), Limnophila (Ancylidae, Physidae, Lymnaeidae), Pulmonata (Elobiidae), Stylommatophor (Helicidae, Punctidae, Succineidae, Vitrinidae, Vertiginidae, Zonitidae), Basommatophora (Lymnaeidae), Caenogastropoda (Semisulcospiridae) are mainly herbivore, epilithic or epiphytic. Because spring water has high transparency and aquatic plants can get sufficient sunlight and nutrients for production, springs can provide habitats for the herbivore species. Numbers of them are commonly found in spring-fed habitats. Species Valvata piscinails (Valvatidae, Heterostropha) is considered to have a high affinity for springs.

Clitellata

There are Enchytraeidae and Propappidae (Enchytraeid) recorded in this class. Marionina argentea is identified as a groundwater-dependent species (Martin et al., 2015). Mesenchytraeus armatus is considered a stenothermal species (Healy and Fend, 2002).

Hirudinea

Hirudinea contains two orders, Arhynchobdellida (Erpobdellidae, Salifidae) and Rhynchobdellida (Glossiphoniidae, Piscicolidae). Most of them are stream dwellers. Species Erpobdella octoculata (Piscicolidae, Rhynchobdellida) is identified as cold stenothermal, which may be used as a spring indicator.

Ostracoda

Species in the Podocopida and Calanoida within the class Ostracoda appear to preferentially inhabit springs and other places where groundwater and surface water mix (e.g., hyporheic zones). There are three families (Candonidae, Cyprididae and Loxoconchidae) in the Podocopida and the family Diaptomidae in the Calanoida. Species in genera Candona, Cryptocandona, and Febaefor- miscandona within the Candonidae, genera Ilyocypris, Cyprinotus, Cyclocypris, Cypris, Chydorus, Eucyclops, Eucypris, Iliocypris, Potamocypris, Psychrodromus, Scottia, Cavernocypris, Stenocypris, Cyclocypris and Heterocypris within the Cyprididae, genus Paradiaptomus (Diaptomidae) and genus Pseudolimnocythere (Loxoconchidae) are considered as spring indicators (Karanovic, 2012).

Maxillopoda

Species in the Maxillopoda tend to dominate springs and groundwater habitats. There are two orders and three families recorded in Maxillopoda, Cyclopoida (Cyclopidae), Harpacticoida (Canthocamptidae, Parastenocarididae). Species in genera Acanthocyclops, Afrocyclops, Diacyclops, Eucyclops, Paracyclops, Megacyclops and Itocyclops within the Cyclopidae, genera Attassaheyella (Attheyella), Bryocamptus (Arcticocamptus), Elaphoidella, Hypocamptus, Moraria and Stygepactophanes within the Canthocamptidae and genus Parastenocaris (Parastenocarididae) are considered freshwater spring indicators.

Malacostraca

Species in the Amphipoda and Isopoda appear to inhabit spring and groundwater habitats (Gooch and Glazier, 1991; Webb et al., 1998). Paraleptamphopiidae (Amphipoda) and Phreatoicidae (Isopoda) contain some groundwater and spring species (Scarsbrook et al., 2007). Sutherland (2005) found most Paraleptamphopus species in small springs and streamside ditches. Scarsbrook and Haase (2003) found surface stream form, possible spring specialist, and unpigmented and eyeless groundwater form from Paraleptamphopus species.

In this study, there are four orders (Amphipoda, Isopoda, Decapoda and Thermosbaenacea) and 19 families recorded in the class Malacostraca. Within the Amphipoda, species in the genera Crangonyx, Synurella, Stygobromus, Mexiweckelia within the Crangonyctidae, genera Crangonyx, Orconectes, Stygobromus, Gammarus, Niphargus within the Niphargidae and genus Hyalella, Ecoiphargus, Niphargus within the Gammaridae and genus Paraleptamphopus (Hadziidae), Pseudocrangonyx (Hylellidae), Eocrangonyx (Mesogammaridae), Sternomoera (Pontogeneiidae) are considered as freshwater spring indicators. Species in genera Echinogammarus and Gammarus within the Crangonyctidae are wetland dwellers. They can be found in some types of springs within the wetlands.

Within the Isopoda, species Proasellus deminutus (Asellidae) is considered a freshwater spring indicator. The remaining species within the Asellidae are identified as lentic species that may inhabit in springs. Most species in the families Atyidae and Cambaridae within the Decapoda and the species Tethysbaena texana within the Monodellidae (Thermosbaenacea) are considered stream dwellers. The species Geothelphusa dehaani (Potamidae, Decapoda) appears to be a cold stenothermal organism that inhabits cold waters in headwaters and springs. It can be a possible indicator of freshwater springs.

Insecta

Insecta species denominate surface water habitats around the world. There appear to be a high-level diversity of orders and families within the class Insecta. 127 families representing 12 orders (Ephemeroptera, Odonata, Plecoptera, Hemiptera, Megaloptera, Neuroptera, Trichoptera, Lepidoptera, Diptera, Coleoptera, Thysanoptera) are reviewed in the class Insecta. Most of them are considered stream dwellers. Some contain spring specialists. Species in the genera Pseudoeconesus larvae and Oeconesus were often found in spring sites (Winterbourn et al., 2000; Scarsbrook and Haase, 2003).

Many species in Insecta can be considered cold stenothermal organisms. Families such as Austroperlidae, Capniidae, Chloroperlidae, Gripopterygidae, Leuctridae, Nemouridae, Notonemouridae, Peltoperlidae, Perlidae, Plecoptera within Plecotera, families Sialidae, Corydalidae, Nannochoristidae, Osmylidae within the Megaloptera, families Philopotamidae, Phryganeidae, Psychomyiidae, Pyralidae, Sericostomatidae within the Trichoptera, families Athericidae, Dixidae, Dolichopodidae, Psychodidae, genus Boreochlus, Rheocricotopusand, Stempellinella, Pentaneurella, Pentaneura, species Metriocnemus fuscipes, Prodiamesa olivacea within the Chironomidae, families Dytiscidae, Hydrophilidae within the Coleoptera, Osmylidae within the Neuroptera, family Calopterygidae within the Odonata, Heptageniidae within the Ephemeroptera tend to appear to preferentially inhibit cold waters They can be found both in headwaters and springs. Several species in Insecta can be identified as herbivora, epilithic or epiphytic according to their ecological

information regarding life types and feeding types. Species in the families Apataniidae, Goeridae, Glossosomatidae and Hydroptilidae within Trichopteradominate many stone-surface habitats both in streams and spring-fed habitats. Ephemeroptera is one of the major components of most surface waters, living in diverse substrata such as stony bottoms, sand, mud, and aquatic vegetation, and most of them are herbivores or detritivores ( Davies and Walker, 2013). Baetidae, Ephemereliidae, Heptageniidae, Leptophlebiidae, Lepidostomatidae, Limnephilidae, Ceratopogonidae, Chironomidae, Empididae are mostly filter feeders and deposit feeders which are often found in streams and are usually associated with aquatic plants. They mainly dominate stream habitats; since streams that functioned as lotic habitats appear to be more suitable than lentic habitats in springs.

Bivalvia

Species in Sphaeriidae (Pelecydoda) and Corbiculidae (Veneroida) are considered as stream dwellers that mainly inhabit surface waters and may appear in springs and spring-fed streams by accident.

Oligochaeta

There are three orders and four families recorded in Oligochaeta: Haplotaxida (Tubificidae, Lumbricidae), Lumbriculida (Lumbriculidae) and Tubificida (Naididae). They are lentic species, which appear to inhabit still waters. They can be found both in streams and spring-fed flows.

Arachnida

Species in the genera Atractides, Hgrobates, Lebertia, Partnunia, Sperchon within the Hygrobatidae (Trombidiformes) can be usually found in lentic habitats. Species in the genera Poroliodes (Neoliodidae), Damaeus (Damaeidae), Cepheus (Cepheidae), Amerus (Ameridae), Xenillus (Xenillidae) and Pilogalumna (Galumnidae) within the Sarcoptiformes can be commonly found in wetland habitats. As some types of springs can be functioning as lentic habitats and wetland habitats, lentic species and wetland species in the Arachnida can be also found in springs. The remaining species in the Arachnida are considered stream dwellers.1.3 Application of spring indicator

1.3 Applicatin of Spring Indicator

It is widely known that the special characteristics of springs, the locations of springs at the interface between groundwater, surface water and terrestrial ecosystems have led to a high value of their biodiversity and high contribution to ecosystem services. However, until recently, very little research has focused on spring ecosystems or their dependent species. This lack of information and attention to spring ecosystems has resulted in the loss of many springs through poor groundwater and land-use practices. The key element in sustainable spring habitat management and conservation is to recognize

the location of springs in freshwater ecosystems (Barquı ́n and Scarsbrook, 2008). To approach this, models and simulations have been developed to estimate the groundwater flow. However, it can be difficult to achieve since running a model or a simulation needs some certain assumptions (e.g. underlying geological data), which makes it rather difficult to estimate groundwater flow on a local scale.

As increasing recognition of the importance of cross-discipline studies and development of groundwater ecology, a new paradigm emphasizes that sustainable groundwater management needs not only abiotic, physicochemical criteria, but also the biological criteria (Tomlinson et al., 2007; Hancock et al., 2005; Boulton et al., 2008). Spring fauna has been suggested as a valuable and cheap indicator for monitoring groundwater quality (Tomlinson et al., 2007; Stein et al., 2010). However, the potential use of spring fauna as biological indicators for the assessment of groundwater contribution is still seldom accessible. In this paper, we suggested a list of spring indicators of benthic invertebrates including three potential bio-indicator groups (groundwater-dependent species, stenothermal species and epiphytic species). Once the benthic community of springs has been sampled, this information can be evaluated using the spring indicator list. Regarding the identification of four potential bio- indicator groups, we have provided a short introduction in this paper; further information should be referred to in related documents. There are a few points that still need to be recognized here regarding the identification and application of stenothermal species and groundwater-dependent species. They are very important groups inside the spring indicators as the result of the taxonomic composition shown in Figure 7. Considering the high diversity of these two groups, the identification and application should vary according to geological and geographical environments.

The stenothermal species refer to benthic invertebrates that have narrow ranges of thermal tolerance. It includes cold-adapted and warm-adapted stenotherms. Respectively, non-stenotherms could be considered as eurytherms which have adapted to a wide range of temperature. However, proper identification of a given species can be difficult and uncertain. There are few studies on thermotolerance and cold hardiness involving aquatic insects and accurate data on the upper and lower thermal limits for many species are still rare (Lencioni and Bernabo`, 2017; Logan and Buckley, 2015). Furthermore, the classification of species into stenotherm or eurytherm group can be variable because of their geographical distribution and their state of relying on or being dependent on spring habitats (Figure 8). Generally, high latitude polar species are considered as cold-adapted stenotherms as they possess the lowest upper thermal threshold of all organisms (Lencioni and Bernabo`, 2017). They can be found in streams and coldwater habitats of low latitude regions. During the summer season, due to their upper thermal limits of heat tolerance, they may find springs or hyporheic zones as suitable habitats. For example, the species B. antarctica from the Antarctic region, Pseudodiamesa branickii and Diamesa cinerella Meigen from alpine streams are investigated as cold stenothermal species for thermotolerance and cold hardiness studies (Rinehart et al., 2006; Bernabo` et al., 2011;

Lencioni et al., 2013). Lencioni and Rossar (2005) have reported the subfamily Diamesinae of Diptera as a dominant taxon in cold alpine streams and springs. On the other hand, some stenothermal polar species could not survive if exposed to freezing temperature or sub-zero environments during winter periods when surrounding water freezes or the water temperature goes beyond their lethal limits of cold hardiness. In high latitude regions, some of these species may find springs or hyporheic zones as a refuge (Lencioni and Spitale, 2015). Within this context, stenothermal species could be considered as spring indicators under certain circumstances. In this study, we have selected some stenothermal species as spring indicators. However, when comes to the application of these stenothermal spring indicators, we need to consider their upper and lower thermal limits, their geographical distribution and their spring dependency. Generally, stenothermal polar species could be used as possible spring indicators in temperate and tropical zones. While in Polar regions, benthic invertebrate species that have adapted to a narrow range of temperatures close to local spring water temperature could be considered as biological indicators of springs. Besides the two spring indicator groups, there are also many species recorded as possible spring indicators in the previous works, but the reasons maintained unknown. Species such as epiphytic species (macroinvertebrates associated with bryophytes) could be considered spring indicators under certain circumstances. Given the stability of springs as habitats, stones with moss mats are more common at spring sites. This could be one of the possible explanations for the abundance of the stone surface and moss mat dwellers at springs and spring-fed streams. Many studies have shown that the epiphytic species prefer cold waters. Tada and Satake (1994) reported epiphytic zoobenthos at the upper reaches of a cool mountain stream. Species Micrasema sp. MC was the most abundant taxon found on bryophyte mats at the headwater sites. Storey and Quinn (2008) found the epiphytic species, Kempynus (Order Neuroptera) in small perennial headwaters. Within this context, epiphytic species share some common characteristics with stenothermal species, thus they can be considered possible spring indicators in some cases. Aquatic invertebrates are numerous and exhibit graded responses to various disturbances in their surrounding environments (Merrit et al., 2008). They have been used as bio-indicators in the river management, for purposes such as water quality assessment and habitat quality evaluation (Harikumar et al., 2014; Scherr, 2010). Hamilton (2013) also studied the effects of climate change on stream invertebrates in their role as biological indicators and responses to disturbance. Some studies have researched macroinvertebrate community composition and distribution in springs, however, it is still unclear why these species have adapted or inhabit in springs. Wigger et al. (2015) have researched spring habitats along an altitudinal gradient of about 2000 m in a valley in the Bernese Alps. The purpose was to explore the corresponding spring fauna with the environmental parameters of springs at different altitudes. We expected more attention to spring habitats and their dependent species. It becomes imperative to explore this issue considering the potential risks of temperature variation threatened by climate change. More efforts should be made on genomic approaches, cold hardiness, and thermotolerance studies to provide additional information on stenothermal species. Investigating the 34

distribution and migration of stenothermal species, researching their thermotolerance and cold hardiness would be great strategies for monitoring the influence of climate change. Such exploring needs more integrative studies and research focused on the development of methods and tools for integrated management of water resource systems combining hydrology, engineering, and ecology.

Figure 8 Scheme of the geographical distribution of spring dependency of stenotherms. Spring

dependency refers to the state of relying on or being dependent on spring habitats. Spring contribution to stenothermal species shows the number of spring habitats contributed to supporting the survival of stenothermal species. Blue and red square indicate a latitudinal range of cold and warm stenothermal species, respectively. Dark shaded range in each square indicates spring dependent zone in each species.

1.4 Conclusion

This study identified spring indicator taxa of benthic invertebrates from data set collected from field research and literature on a continental and worldwide scale. The information provided by the spring indicator taxa of an aquifer can reflect the contacts between the surface origins of the water and may be used as an indicator of groundwater and spring evaluation. Given the susceptibility of springs and groundwater to modification from land-use change and water abstraction, many species should be considered at risk, and urgent attention is required to incorporate biological indicators into an assessment of groundwater and spring management. Many factors can be considered during a decision-making policy setting. Applying biological indicators to conservation work is essential for sustainable river ecosystem management and the well-being of human life.

CHAPTER 2 Classification of Riverine Spring Habitats and Fauna Characteristics

There are several different classifications of springs. A wide variety of spring types and several physical classification schemes have been developed. The spring classification system includes geology, hydrology, physical, chemical, and biological parameters at the spring head and within the spring flow. Springs are classified based on its geologic origin, rock type, physical properties, chemical composition, discharge rates, groundwater flow path, etc.

In this chapter, we proposed a new classification of spring types, focusing on the locations of springs in relation to the main channel of rivers based on differences in water levels. The classifications put forward by this paper stress the different meanings of spring as habitats for organisms.

2.1 Spring Classification in this Study We firstly characterize springs by their horizontal distribution in relation to the mainstream channel

and divided springs into two types (Figure 9).

Spring-flow type: the spring emerges from outside of the one-year-floodplain zone, forming spring flows into a mainstream channel.

Riverbed spring type: the spring emerges within the riverbed. In this paper, the riverbed is defined as areas within the one-year-floodplain zone.

Figure 9 Sketches of horizontal distribution of springs in relation to the stream channel. (1) Spring- flow type (a), (2) Riverbed spring type: b. Floodplain spring, c. Water's edge spring and d. Under- water spring. S=spring source; W=wetland.

The characteristic of spring-flow type is that the spring source is located outside the area where flood event occurs. The head source of springs, as well as parts of the spring flow, are not disturbed by surface water from the river. Riverbed spring-type located inside of the one-year-floodplain zone would be inundated by surface water from the main channel when a flood event happens. One-year- floodplain zone is defined as the area that will be inundated by flood events once a year on average; the water level during the flood events is defined as high water level (HWL), the water level during the drought season is referred to as low water level (LWL). The Riverbed spring is classified into three types according to the relative height of spring to the water level of the mainstream channel:

Floodplain spring: spring emerges within the upper zone of the riverbed.

Water’s edge spring: spring emerges on the water’s edge at low-water-level.

Under-water spring: spring emerges under the water in the mainstream channel.

In the classification, the location of the spring-flow type is higher than the high water level of the main channel (Figure 10). Floodplain spring is lower than the high water level. Water's edge spring is closed to the low water level of the main channel, and the under-water spring is at the bottom of the river.

Figure 10 Sketches of riverbed spring types based on the vertical location of springs in relation to channel water level: a. Spring-flow type, b. Floodplain spring, c. Water's edge spring, and d. Under- water spring. G=groundwater; S=spring source; HWL=high water level; LWL=low water level.

2.2 Study Sites and Data Collection Historical observed data collected at four research sites were used to characterize taxonomic composition, diversity, and ecological types of benthic invertebrates in each spring habitat type (Table 2). The observed benthic invertebrate data were collected from field research (Gamada River, 2015) and previous work of Nomura (2007) and Suzuki (2007). Benthic invertebrate samples were collected using a D-frame net. Kicking and sweeping were used at muddy and gravel habitats. Kick and sweep are highly versatile techniques; it is widely used on rock, sand, gravel and mud bottoms (Abdelsalam and Tanida, 2013). The sampling methods used by the US state agencies for collecting macroinvertebrates are surveyed and the results showed that kick sampling was the widest technique that used in fieldwork with D-frame nets as tools(Carter and Resh, 2001). Collected benthic invertebrates with organic particles were transferred into PVC bottles and bags, sorted and preserved in ethyl alcohol. The benthic invertebrate samples were brought back to the laboratory after the fieldwork. The specimens were sorted and identified to the lowest possible level using relevant references.

Table 2 Physicochemical variables at each research site.

Westside of Eastside of

Hodakanomori, Kamikamo Shrine, Kamikamo Shrine, Nebori Valley Hiru Valley Kamo River Kamo River

Spring Type Spring-flow type Floodplain spring Water's edge spring Under-water spring

WT(℃) 8.08 ± 1.33 7.92 ± 1.86 10.00 ± 0.50 12

EC (ms/cm) 36.70 ± 4.00 53.00±8.00 - -

DO (%) 81.06 ± 1.73 100.20 ± 16.60 73.5 ± 23.3

DO (mg/L) 9.58 ± 0.47 11.20 ± 1.60 7.01 ± 3.57

Field research, Reference Nomura (2007) 2015 Suzuki (2007)

(1) Spring-Flow Type: Hodakanomori, Gamada River

Figure 11 The research site investigated by Nomura (2007) in Hodakanomori, Nobori Valley, Gamada River represents the spring-flow type (Photo by Takemon, 2007).

Gamata River (蒲田川, Gamata-gawa) is the upper reaches of River Jinzū basin. It has its source at Hidarimata Valley and Minimata Valley in the eastern part of Gifu Prefecture, Japan, and joins with the Takahara River. Nomura (2007) researched habitat conditions and invertebrate communities of six

valleys in the Gamada basin. Table 3 shows the geographic characteristics of valleys in the Gamata basin, in terms of the catchment area. Among these valleys, Nebori is the valley where researchers found springs emerging from a forest called Hodakanomori (Figure 11). The springs emerge from the forest located outside of the floodplain zone and flow into the main river channel. The benthic invertebrate data collected by Nomura were used for analyzing fauna characteristics of the spring- flow type.

Table 3 The geographic characteristics of valleys in the Gamata basin (Nomura, 2007).

Nebori Hiru Wari Sodega Shiromizu Kuro

Valley Valley Valley Valley Valley Valley

Catchment area (km2) 0.84 0.85 1.08 6.21 2.44 1.43

Bare area (km2) 0.00 0.00 0.05 0.29 0.59 0.43

Collapse area (%) 0.00 0.03 4.69 4.59 24.16 30.05

Number of erosion control dams(including those under 0.00 1.00 9.00 16.00 2.00 2.00 construction)

Konabe Koito Kuriya Karukaya Hora Valley Valley Valley Valley

Cathment area (km2) 4.92 1.90 3.88 0.55 2.34

Bare area (km2) 0.87 0.07 0.09 0.00 0.12

Collapse area (%) 17.71 3.82 2.23 0.56 4.94

Number of erosion control dams(including those under 10(3) 4.00 0.00 0.00 7.00 construction)

(2) Floodplain Spring: Hiru Valley, Gamada River

We researched six sites at four valleys in Gamada River in September 2015 (Figure 13). Physico- chemical parameters were measured at study sites using a water quality monitor (Table 2), and benthic invertebrate samples were collected using D-frame net-tools, kicking and sweeping methods (Appendix 2). In Hiru Valley, we researched a spring within the upper zone of the riverbed (Figure 12). Benthic invertebrate data collected in Hiru Valley were used to show the ecological types in the floodplain spring.

Figure 12 Terrestrial spring: the research site at Hiru Valley (Photo by Yamashiki, 2015).

Figure 13 Study sites of Gamata River (Gifu, Japan): Nebori Valley (S1 and S2), Sodega Valley (S3), Hiru Valley (S4 and S5), and Wari Valley (S6).

(3) Water’s Edge Spring: West Side of Kamogamo Shrine, Kamo River

Kamo River has its source at the area of Mount Sajikigatake in the northern ward of Kyoto. It flows into the Kyoto Basin and joins with the Takano River. Suzuki (2007) researched benthic invertebrate assemblages at the west side of the Kamigamo shrine and east side of the Shimogamo shrine in the Kamo River. At the westside of the Kamigamo shrine, springs were found emerging from the west edge of the Kamo River (Figure 14). The invertebrate data collected at this site were used in the analysis.

Figure 14 Water’s edge spring: the research site located at the westside of Kamigamo Shrine (Photo by Takemon, 2016).

(4) Under-Water Spring: East Side of Shimogamo Shrine, Kamo River

According to Suzuki (2007), springs were emerging from the bottom of the river on the east side of the Shimogamo shrine, Kamo River (Figure 15). We used the invertebrate data of this research site for analyzing the characteristic of under-water spring.

Figure 15 Under-water spring: the research site at the east side of Shimogamo Shrine, Kamo River (Photo by Takemon, 2007).

2.3 Data Analysis We analyzed the taxonomic composition, diversity, and ecological types of benthic invertebrate samples collected in each spring habitat type using the historical observed data. The analysis of ecological types in each spring type used correspondence analysis (CA) to understand the interactions between benthic invertebrates and spring microhabitats.

Ecological types are developed based on the theory of evolutionary and the concepts of adaptive radiation and adaptive convergence. The benthic invertebrates are strongly affected by their environments, including sediment composition and quality, water quality, and hydrological factors that influence the physical habitats. Because they are so dependent on their surroundings, the ecological types of benthic invertebrates reflect the conditions of the aquatic environment. According to the work of Takemon (2005), the ecological types include habitat type, substrate type, life type, functional feeding group, and food type (Table 4). Habitat type describes the general conditions of habitats. Hygropetric habitats are rocks or stones near the water's edge, wetting with the freshwater of rapids and waterfalls. This type of habitat is characterized by special hygrometric species that have adapted to the moist environment. Lotic water type refers to the dynamic water habitats with any flow speed. Lentic water type refers to the still water body in the river system.

Substrate type refers to the bottom substrate in the stream. We classified substrate type into eight

categories: i.e., rock, stone, gravel, sand, mud, litters, plant body and woody debris. Life type is defined as indexes that reflect physical and hydrogeological characteristics of habitats, physical characteristics and life types of benthic invertebrates. Life types are divided into four groups: swimmers (e.g., Baetidae), attachers (e.g., Blepharoceridae, Simuliidae), crawlers (e.g., Plecoptera, Rhyacophilidae, Corydalidae), and burrowers (e.g., Paraleptophlebia, Potamanthodes formosus, Ephemeridae, Gomphidae, Tanytarsus, Microtendipes).

The functional feeding group is a classification of organisms based on methods for food acquisition: grazers, collector-gatherers, shredders, filter-feeders, and predators. Grazers appear to graze and feed on the surface of stones and rocks (e.g., Epeorus latifolium, Epeorus napaeus, Epeorus ikanonis, Eperorus curvatulus), and collector-gatherers are benthos community which collects and feeds on Fine Particle Organic Matter (e.g., Cincticostella nigra, Paraleptophlebia japonica, Choroterpes altioculus, Acentrella gnom). Shredders are observed feed on fallen leaves, aquatic plants. Filter-feeders feed on FPOM and biomass of microorganisms.

Ecological types of the four research sites were summarized into a two-way contingency table containing 15 rows representing ecological types (matrix type, life type, functional feeding type, and food type) and four columns representing four spring habitat types. Each cell of the table recorded the percentage of species associated with ecological type within each spring habitat (Table 5).

The table of ecological types of the four types of spring habitats was submitted to CA to graphically

represent the degree of association between the spring habitats and their ecological types according to their Chi-squared distances. Since we have applied a three-column table to CA which derives two dimensions (Figure 18), the principal inertias (eigenvalues) cover 86.9% of the total variance (Dimension 1 explains 62.7%, Dimension 2 explains 24.2%). Correspondence analysis (CA) was used to summarize the major ecological type pattern within the spring habitat variations. Species richness was calculated at spring type. The Shannon Wiener diversity index and Pielou's evenness index were used to evaluate the species diversity in the samples.

Table 4 Ecological types of freshwater benthic invertebrates in the river system (FPOM: Fine Particulate Organic Matter, particle size >1mm; COPM: Coarse Particulate Organic Matter, particle size: 0.7μm~1mm).

Habitat type Substrate type Life type Functional feeding Food type

group

Lotic habitat Rock Attachers Grazers Herbivore

Lentic habitat Stone Crawlers Collector-gatherers FPOM feeders

Hygropetric Gravel Swimmers Filter-feeders CPOM feeders

habitat Sand Burrowers Shredders Carnivora

Litters Predators Scavenger

Plant body Suckers

Woody debris

Table 5 Contingency table of species percentage that was associated with ecological types at four different spring habitats (The bold items represent the percentage of benthic invertebrate species that ecological types showed significant relations to the research sites in the later Correspondence Analysis).

Ecological type Type a Type b Type c Type d

St 78.57% 69.70% 46.30% 81.48%

Sa 12.86% 9.38% 21.21% 14.81%

Matrix type Pl 8.57% 18.75% 3.03% 3.70%

SWM 17.14% 9.38% 15.15% 9.26%

CRL 48.57% 68.75% 51.52% 59.26%

BRW 21.43% 9.38% 21.21% 24.07%

Life type AT C 17.14% 12.50% 12.12% 7.41%

GR 24.29% 31.25% 27.27% 25.93%

PR 28.57% 21.88% 18.18% 27.78%

FL 8.57% 18.75% 18.18% 14.81%

SH 14.29% 15.63% 6.06% 7.41%

Functional feeding group CG 24.29% 12.50% 30.30% 24.07%

HERB 22.86% 28.13% 21.21% 24.07%

FPOM 30.00% 31.25% 51.52% 40.74%

CARN 32.86% 25.00% 18.18% 24.07%

CPOM 12.86% 15.63% 6.06% 7.41%

Food type SCAV 1.43% 0.00% 3.03% 3.70%

2.4 Biological Differences Among Spring Habitat Types

(1) Taxonomic Composition

According to the historical observed data, a total of 1791 specimens, representing 70 taxa and 68 families of benthic invertebrates were identified in the Hodakanomori (the spring-flow type, Type a) (Table 6). We classified the sampling data and found 11.42% of the spring indicator taxa of stenothermal species in their total taxa composition. The stenothermal species are in the Order of Trichoptera and Diptera (Dolophilodes sp., Rhyacophila lezeyi, Rhyacophila towadensis, Rhyacophila shikotsuensis, Rhyacophila sp., Pagastia sp., Pericoma sp., Dugesia sp.).

In the Hiru valley (the floodplain spring, Type b), we collected invertebrate samples of 238 specimens, representing 36 taxa and 32 families. 11.11% of the total taxa were identified as stenothermal species, including species in the Order of Plecoptera, Trichoptera and Diptera (Yoraper la uenoi, Wormaldia sp., Rhyacophila sp., Pagastia sp.).

On the west side of Kamigamo Shrine, Kamo River (the water-edge spring, Type c), we analyzed historical observed benthic invertebrate data of 827 specimens, representing 39 taxa and 33 families. Stenothermal species in the Order of Diptera (Dixa nipponica) and groundwater-dependent species (Paratya compressa compressa) were found in this site.

On the east side of Shimogamo Shrine, Kamo river (the under-water spring, Type d), we identified historical data including 2254 specimens representing 64 taxa and 39 families of invertebrates. We found that stenothermal species in the Order of Trichoptera and Diptera (Rhyacophila nigrocephala, Dixa nipponica, Pagastia sp., Geothelphusa dehaani) and groundwater-dependent species in the Order of Ostracoda and Amphipoda (Ostracoda gen. spp., Sternomoera sp.) were researched from this site.

Additionally, several species were found to be associated with multiple spring habitat types. For example, the stenothermal species Pagastia sp. were found in spring-flow type, floodplain spring and under-water spring; Rhyacophila sp. are found in spring-flow type and floodplain spring, and Dixa nipponica are found in water-edge spring and under-water spring.

Table 6 Taxonomic composition in each research site.

Study site Spring Taxonomic Composition of spring indicator Percentage habitat composition taxa of total type taxa

Hodakanomori Spring- 1791 A total of 8 taxa of stenothermal 11.42% flow type specimens, species in the Order of Trichoptera 70 taxa, 68 and Diptera (Dolophilodes sp., families Rhyacophila lezeyi, Rhyacophila towadensis, Rhyacophila shikotsuensis, Rhyacophila sp, Pagastia sp., Pericoma sp., Dugesia sp).

Hiru Valley Floodplain 238 A total of 4 taxa of stenothermal 11.11% spring specimens, species in the Order of Plecoptera, 36 taxa, 32 Trichoptera and Diptera (Yoraperla families uenoi, Wormaldia sp., Rhyacophila

sp., Pagastia sp.).

Westside of Water- 827 Dixa nipponica (Diptera) identified 5.13% Kamigamo edge specimens, as stenothermal species and Shrine, Kamo spring 39 taxa, 33 groundwater-dependent species of River families Paratya compressa were found at this site.

Eastside of Under- 2254 A total of 5 taxa of stenothermal 7.81% Kamigamo water specimens, species in the Order of Trichoptera Shrine, Kamo spring 64 taxa, 39 and Diptera were found River families (Rhyacophila nigrocephala, Dixa nipponica, Pagastia sp., Geothelphusa dehaani) and 5 taxa of groundwater-dependent species in the Order of Ostracoda and Amphipoda were found (Ostracoda gen. spp., Sternomoera sp.).

(2) The Patterns of Invertebrate Diversity

The patterns of abundance, species richness, diversity and evenness of benthic invertebrates varied among the four researched spring habitat types (Figure 16). The abundance and species richness are higher at spring-flow type (Type a) and under-water spring (Type d), and lower at floodplain spring (Type b) and water's edge spring (Type c). The richness index values varied from the lowest value of 36 at floodplain spring to the highest value of 64 at under-water spring. The Shannon Wiener diversity index and Pielou's evenness index remain similar at the spring-flow type and floodplain spring, higher than water's edge spring and under-water spring. The Shannon Wiener index varied from the lower values of 1.83 at water's edge spring and 2.50 at under-water spring to the higher values of 3.10 at the spring-flow type and 3.00 at floodplain spring. The Pielou index varied from 0.50 at water's edge spring and 0.55 at Under-water spring to 0.75 at spring-flow spring and 0.84 at floodplain spring.

Figure 16 Benthic invertebrate abundance, species richness, Shannon-Wiener diversity index and Pielou's evenness index in the 4 spring sites (Type a: Spring-flow type, Hodakanomori); Type b: Floodplain spring, Hiru Valley); Type c: Water’s edge spring, West side of Kamigamo Shrine, Kamo River; Type d: Under-water spring, East side of Shimogamo Shrine, Kamo River).

(3) The Patterns of Ecological Types

The results of ecological type analysis showed that in the Hodakanomori site, there was a relatively higher percentage (78.57%) of benthic invertebrates that prefer to inhibit on stones than other matrix types (Figure 17). 48.57% of the total taxa were defined as crawlers in life type index. 28.57% of total taxa were identified as predators in the functional feeding group and 32.86% of benthic invertebrate data were defined as carnivora in the food type.

In Hiru Valley, the biological sample analysis in ecological types showed that the study site was respectively related to matrix type of stones (69.70%), life type of crawlers (68.75%), functional feeding group of grazers (31.25%), and FPOM feeders (31.25%).

On the west side of Kamigamo shrine, Kamo River, the site has 46.30% of stones as the matrix type, respectively higher than other matrix types. The life type, functional feeding group and food type indexes showed that this site was characterized by crawlers (51.52% of total taxa in the life type), collect-gatherers (30.30% of total taxa in the functional feeding group), and FPOM feeders (51.52% of total taxa in the food type).

The results of ecological type analysis at the east side of Shimogamo shrine showed that the spring provided more habitats related to matrix type of stones (81.48%). The life type, functional feeding group and food type identification results showed that this site was more related to crawlers (59.26%),

predators (27.78%), and FPOM feeders (40.74%).

All spring habitats were found a high percentage of macroinvertebrates that prefer matrix types of stones and life types of crawlers. Correspondingly, the percentage of plants is higher in floodplain spring and spring-flow type. Swimmers, attachers, predators and carnivores are more commonly found in spring-flow type. Crawlers, grazers, filter-feeders, shredders, herbivores, and CPOM feeders are more commonly found in floodplain spring. The percentage of collect-gathers and FPOM feeders are respectively higher in water's edge spring. The percentage of burrowers and scavengers are comparatively higher than those of others.

Figure 17 Ecological type composition in four spring habitats (Matrix type: Stones/St, Sands/Sa and Plants/Pl; Life type: Swimmers/SWM, Crawlers/CRL, Burrowers/BRW and Attachers/ATC; Functional feeding group: Grazers/GR, Predators/PR, Filter-feeders/FL, Shredders/SH and Collector- gatherers/CG). 53

Figure 18 Ordination diagrams of CA analysis on ecological types (Matrix type: Stones/St, Sands/Sa and Plants/Pl; Life type: Swimmers/SWM, Crawlers/CRL, Burrowers/BRW and Attachers/ATC;

Functional feeding group: Grazers/GR, Predators/PR, Filter-feeders/FL, Shredders/SH and Collector- gatherers/CG) in relation to four types of spring habitats (Type a: Spring-flow type, Hodakanomori); Type b: Floodplain spring, Hiru Valley); Type c: Water's edge spring, West side of Kamigamo Shrine, Kamo River; Type d: Under-water spring, East side of Shimogomo Shrine, Kamo River).

2.5 Discussion Previous studies reported that spring water acts as a hotspot for aquatic biodiversity and productivity in river ecosystems, because springs are connections of groundwater systems and interface water systems (Cantonati et al., 2006; Scarsbrook et al., 2007). In this study, we focused on developing and understanding several criteria of spring habitat classification and their characteristics of benthic invertebrate communities, which can be fruitfully applied to the answering the question regarding the distinct ecological roles that springs play in a riverine system. We provide a typology of spring habitats within the riverine system. The new method classifies spring habitats based on the locations of springs in relation to the main river channels. In previous spring classifications, springs flowing into one or more stream channels were defined as Rheocrene; springs emerging from low gradient wetlands were defined as Helocrene; and springs emerging from hillslopes were identified as Hillslope (Bornhauser, 1913; Hynes, 1970; Grasby & Londry, 2007; Springer et al., 2008). However, we classified spring types not only based on their geological locations but also their locations in relation to the main river channels and floodplain elements. We divided springs into two groups at a river landscape scale, dividing Rheocrene spring-type into two groups: spring flow type, located outside of the floodplain zone and riverbed spring type, located within the floodplain zone. In addition, both spring types may contain Helocrene and Hillslope which emerge from wetlands and hillslopes but may have different levels of interaction with surface water.

Barquin and Scarsbrook (2008) pointed out that the interaction between groundwater and surface water influences the structure and function of spring habitats, and our results showed that different types of spring habitat have different characteristics of benthic invertebrates. In contrast to the study which suggests species diversity of invertebrates may increase from groundwater to surface spring habitats to downstream sites ( Likens, 2009), we found lower levels of diversity at sites that were closer to rivers than spring sites. Such a pattern may reflect the role of flood disturbance as a constraint to invertebrate diversity (Ward et al., 2002). Gray et al. (2006) investigated braided channels, springs and hillslope streams in the Waimakariri River and the results showed that spring systems embedded within the floodplain zone have respectively high invertebrate biodiversity. Our results showed that springs emerging in the upper zone within the floodplain zone (Floodplain spring) appeared to have higher diversity than those emerging in the lower zone within the floodplain zone (Water’s edge spring), and springs emerging under the water in the main river channel (Under-water spring). The intermediate levels of disturbance may be responsible for explaining the greatest diversity observed in spring within the upper zone of the riverbed. Consequently, our study suggests that spring habitat types with different degrees of connectivity with surface water have different levels of biodiversity and richness. This supports the theory that floods may reduce invertebrate diversity (Burgherr et al., 2002; Reckendorfer et al., 2006) in the case of spring within the lower zone of the floodplain zone and spring emerging under the river. Digby (1999) found that secondary production

within a braided river increased as habitat stability increased, and spring habitats located relatively close to river channels may have unstable environmental conditions in which strong surface water disturbances constrain the invertebrate diversity. However, there is also another circumstance in which flood disturbances may contribute to invertebrate diversity. In this study, we found that springs that emerge within the upper zone of the one-year-floodplain zone showed the highest variety of benthic invertebrates. The results indicated that flood disturbance plays an important role in the arrangement of invertebrate diversity patterns.

Furthermore, our Correspondence Analysis (CA) on ecological types of benthic invertebrates at each research site has contributed to the extension of the findings of previous observations. The results suggest that invertebrate communities are not only distinguishable in species diversity and richness, but also vary in terms of their ecological types (e.g. lifestyle and food types). As previous studies suggested that many invertebrates have morphological, behavioral and life-history strategies to cope with flood effects (Rempel et al., 1999; Sedell, et al., 1990; Townsend & Hildrew 1994), invertebrates also appear to have different life strategies to adapt to their environment. Spring-flow type appeared to have more interaction with stones, swimmers, attachers, predators, and Carnivora. Spring-flow type is the spring habitat located outside of the floodplain zone and therefore is likely to be stable and has stony and rocky substrates. The nutrients of spring water promote the growth of plants and algae. Swimmers adapt to swimming in lotic and lentic habitats. Attachers prefer stone surface, stony and rocky substrates, feeding on plants and algae. Predators are carnivores that adapt to the environment where FPOM is scarce. The Floodplain spring emerging in the upper zone of the riverbed share similar ecological patterns with spring-flow type. Both spring-flow type and floodplain spring have a high percentage of plants, shredders and CPOM feeders. Shredders feed on plants and leaves. COPM is more common in the up-stream with trees and plants. Water's edge spring and under- water spring located relatively close to the main river channel seem to have more interactions with surface water and more disturbances than the other spring habitats. These two spring types share similarities in their patterns of ecological types. Both water's edge springs and under-water springs respectively are characterized by matrix type of sands. Their invertebrate communities include burrowers that live in the interspace of sands and gravels, collect-gatherers which feed on algae and FPOM. The results of the Correspondence Analysis indicated that substrate types (rocks, stones, sands, and plants), food types, life types and the types of functional feeding groups were responsible for discriminating invertebrate assemblages in the four different spring habitats. Spring contribution (spring size and spring permanence), habitat stability, water quality and the relations to surface water might be the factors most responsible for explaining the different patterns of ecological types in the four spring habitats.

Riverine springs and spring-fed streams are respectively regarded as habitats for some characteristic aquatic organisms. Small spring-fed channels and backwaters are found as sites of high invertebrate

productivity and diversity (Pierce 1979; Hughey 1989). Gray et al. (2006) pointed out the view of braided rivers being unstable habitats with low species diversity may not "take into account the diverse array of non-braided aquatic habitats present in their floodplains". In this study, we identified the distinct hydrogeological patterns in the arrangement of spring macro-habitats within a riverine system. Our results demonstrate the importance of spring habitat diversity and their varying degrees of interactions with surface water in structuring invertebrate diversity and ecological patterns. Our thinking about the ecological role of springs and macro-habitats within the riverine system should no doubt be grounded in how we conceive the notion of basin management itself. Studies of braided rivers suggest that braided river management should consider habitat diversity and the dynamic mosaic of interconnected habitats that characterize the river (Digby, 1999; Arscott et al., 2005). Consequently, how we think about each spring type in river management is likely related to our understanding of two relationships:

a. The hydrogeological context of springs related to river system;

b. The ecological role of spring habitats in the river ecosystem.

Table 7 shows our ideas of thinking on riverine springs across these two dimensions. The rows represent the spring types which were classified according to their hydrogeological structure, where spring types differed in their interactions with main river channels (surface water). The columns show

the ecological functions of springs within the riverine system: accessibility as a refuge, habitat stability and habitat suitability for stygophile. We use the information provided in Table 7 to highlight the importance of identifying distinct micro-habitats of riverine springs. The locations of spring types in relation to the main river channel water level may influence the functions of springs as habitats for aquatic organisms. Environmental conditions in the springs are relatively stable. Thus, the spring-flow type is considered to provide a more stable and suitable habitat for stygophile and other unique aquatic species. In addition, springs are stable in water flow, sediment dynamics and disturbance through flood seasons and supra-seasonal drought (Stubbington & Wood 2013). Springs and spring- fed streams can function as refuges for some aquatic organisms during flood seasons and supra- seasonal droughts. Springwater also has relatively long-term, stable temperatures of water which makes it a suitable habitat and refuge for cold stenothermal organisms to survive climate changes. The accessibility as a refuge for aquatic animals may vary according to different patterns. In this case, water's edge springs and under-water springs which are closer to the main river channel may have more accessibility as refuges for aquatic animals.

Table 7 Habitat function of each spring type.

Accessibility Habitat suitability Spring type Habitat stability as refuge for stygophiles

a. Spring-flow type ～＋＋＋＋＋＋＋＋

b. Floodplain spring ＋＋＋＋

c. Water’s edge spring ＋＋＋

d. Under-water spring ＋＋＋＋

Different types of spring habitats may have different ecological roles and interactions with benthic invertebrates. One well-understood conception of how river and spring management provides goods and services is that many factors and multi-stakeholder relations engage one another in river management practice, and the resulting compromise reflects the best possible balancing of conflicting demands. In this system, experts and researchers would best serve this decision making progress by identifying the ecological roles of different spring types in the form of policy alternatives to decision-

makers who can then set up the spring and river management priorities among different courses of conservation plans.

It is important to recognize different types of spring habitats associated with the dynamics of floodplain elements for structuring the spring-associated benthic assemblages. We suggest that river management would benefit greatly from the understanding of how different habitats associated with aquatic biodiversity patterns (and their functional grouping pattern), to obtain more information about ecological processes for predicting the effects of long-term environmental changes. To predict the effects of such changes, models are required but there are some circumstances in which models cannot provide detailed information. The field observation research spanning many years constantly faces challenges.

2.6 Conclusion We provide a classification of spring habitats within the braided river landscape, which can be useful in identifying biodiversity values across different spring habitats of a braided riverscape. The spring types are classified based on the locations of springs in relation to the main river channels. This helped to demonstrate the importance of different types of spring habitats associated with the dynamics of flood-plain elements for structuring the biodiversity and ecological types of spring- associated benthic assemblages. We hope local leaders and stakeholders can recognize the dynamic interactions between springs and surface water in the arrangement of riverine spring habitats, and their important consequences for biodiversity and ecological types of invertebrates. Accordingly, appropriate decisions and governance frameworks underpinning management practices can be made according to the local context.

CHAPTER 3 Application to Conservation of Spring Ecosystems and Environmental Education

Spring habitats present as a mosaic of heterogeneous, connected micro-habitats in our landscape. Despite the small area of springs, spring habitats support many characteristic creatures. Sun et al (2018) showed that the spatial location of springs among groundwater, surface water and terrestrial ecosystems has an important role in shaping species composition and spatial distribution of benthic invertebrate communities. The location and geomorphological structure of spring habitat are crucially important also for cold stenothermal fishes to survive under high water temperature conditions in temperate streams. Cold stenothermal fishes such as salmonids (Naito et al, 2012) and Ayu fish (Nakagawa et al, 2015) are increasingly threatened by climate change. As water temperature rises, they lost their coldwater habitats and must adapt or migrate to other habitats. Springs and spring-fed streams provide the cold stenothermal species refuges to survive as the water temperature becomes warm. Consequently, instream springs play an important role in biodiversity conservation under global warming conditions.

To conserve the spring ecosystems sustainably, monitoring and assessment of springs are

indispensable. Management and land-use practices without sufficient information and attention to spring ecosystems may lead to significant loss of springs, biodiversity and ecological functions for human benefits. To promote integrated researches and management policies, in section 3.1 and section 3.2, we combine information on spring monitoring and assessment approaches. For the spring-scale approach, we suggest the use of spring indicators and spring habitat classification.

In the case of springs connected with surface water stream flows, the ecological function of springs will change with the size and depth of the spring waterbody in comparison with the stream waterbody. For predicting the thermal regime changes of the spring ecosystems with changing size and depth of the waterbody, the numerical model approach is useful particularly for assessing the human impact on the ecosystems. In this section, we provide information on spring researches in the Lake Biwa basin and suggest that more attention should be paid on the integrated lake management contributed to spring monitoring.

As providing such information and suggestions to decision-makers are essential to set up the conservation priorities in ecosystem management practice, environmental education serves to raise public environmental awareness. In section 3.3, the advice is given for the researches engaging in environmental education, and an environmental education project provides an extended example of the action research process.

3.1 Spring Monitoring

Several direct (field measurements) and indirect methods (models and simulations) of groundwater monitoring bring great benefits to spring management, With the consideration of biological factors associated with the environments, springs and groundwater can be monitored on a local scale. Aquatic invertebrates are numerous and exhibit graded responses to various disturbances in their surrounding environments (Merrit et al. 2008). Because they are closely dependent on their surrounding environments, the invertebrates serve as biological indicators of a variety of environments. They have been used as bio-indicators in river management, for purposes such as water quality assessment and habitat quality evaluation (Harikumar et al. 2014; Scherr, 2010). Hamilton and Anna (2013) also studied the effects of climate change on stream invertebrates in their role as biological indicators and responses to disturbances. Springs are considered as a hotspot of biodiversity. There are some characterized invertebrate assembles inhabiting in the springs. We identified spring indicator taxa of benthic invertebrates from data sets collected from field researches and literature on a continental and world-wide scale (Chapter 1). The information provided by the spring indicator taxa of an aquifer can reflect the contacts between surface water and groundwater at a certain point compared to hydrochemical data indicating the origin of the water and may be used as indicators of groundwater and springs management. Collecting invertebrate samples and analyze the composition using the spring indicator list can provide information on the spring identification in a

spring-scale. Monitoring the invertebrate communities can also help us to access the stability and health of springs.

Field measurements require time and energy to capture sufficient data for estimation, but it is difficult for some field researches which is operated under limited conditions within a limited time. Models and simulations can make a basin-scale scientific estimation of water flows within a region based on the data analysis and some certain assumptions.

Lake Biwa is the largest freshwater lake in Japan, which locates in Shiga Prefecture. Groundwater and springs are found in the alluvial fan, delta regions at the western shore and the bottom of the lake (Kobayasi and Tatumi, 1999). The main source of local groundwater and springs are considered as the snowmelt of Hira Mountain (Horiuchi and Kobayashi, 2008). Several researches have been conducted to investigate the physicochemical characteristics of springs and their relationship with aquatic fishes and plants at the shore (Kobayashi and Tatumi, 1999; Horiuchi and Kobayashi, 2008; Takahashi and Asaeda, 2014). Nakayama et al. (2000) have evaluated the groundwater flow system in the Lake Biwa basin by analyzing the stable isotope ratios in precipitation and groundwater. However, there are few studies focused on spring monitoring in the lake basin. There are several studies conducted to estimate the nutrient and ecosystem cycling in Lake Biwa using models and simulations (Okugawa and Somiya, 1983; Somiya et al., 1993).

Yamashiki et al. (2003) developed a 3D modeling approach for Lake Biwa basin management, which enables calculation in the water temperature distribution, flow rates and concentrations of pollutants and organic simulation conditions. By simulating the physicochemical variables (eg. water temperature and changes) of the lake, the status of springs in the lake basin can be estimated by the model. For example, during the winter season, there is usually little effective circulation in a frozen lake (Figure 20). However, springs emerging from the bottom of the lake are considered to have a respectively higher temperature than surrounding bottom water. As a result, the warm spring water will rise gradually because of their smaller density, vertical convection thus takes place. In the case of an unfrozen lake (e.g., Lake Biwa), there are circulations during the winter season. Spring flows and springs emerging from the bottom of the lake are more likely to rise or mix with the lake water. During the summer, the lake is stratified and has a surface water layer (epilimnion) with a higher temperature than spring water and the surrounding temperature at thermocline might be a similar temperature (Figure 19). Springs emerging from the bottom of the lake are more likely to rise and mix with the water in hypolimnion and thermocline. This can be numerically simulated under the assumption of temperature profile around the spring. To estimate the spring flows in a lake, we use the Biwa 3D model to simulate the water temperature change in Lake Biwa during the winter season (Figure 21). We assume that spring waters (water temperature: 14℃) flow into the westside (Ado River mouth: 35°19'N, 136°04'E) and eastside (Maibara section: 35°19'N, 136°16'E) of the lake with river discharges. The discharges of spring water are calculated according to their proportion of river water. The simulation is run from January to March, using the input data include the monthly water temperature and river discharge data of 2002. In January, the lake is separated into two temperature zone. The water temperature at the lake bottom is lower than the upper zone. After we run the model, the water temperature of Lake Biwa is gradually changed. The final simulation result shows that the separated temperature zone disappeared in March. The result indicates that there might be circulation occurred in the lake which mixes the upper zone water and bottom water. There are clear spring flows shown on the sides of the lake. The east side has more spring flow discharge than the west side. This is probably because the river discharge of the east side is larger than the west. On both sides, we found most of the spring flows tend to stay in the upper zone, only litter spring water flowing into the lake bottom. Springwaters tend to rise or stay in the upper zone of the lake, probably because they have a smaller density. Further approaches are needed to grope for the application possibility of lake modeling in the spring monitoring.

Groundwater discharges to surface water in terms of springs. In many places, springs can be found in streams and lakes, providing water, nutrients and stable water temperature. To achieve sustainable spring management, we need to pay attention to the connections between springs and other water bodies, integrating spring conservation into river and lake management framework. Spring monitoring and assessment should also consider approaches from different scales. In Chapter 2, we emphasized

the importance of identifying different riverine spring types, integrating spring conservation with river management. We suggest that more attention should be paid on the further step ahead towards approaches integrating spring monitoring with lake basin management.

Figure 19 The circulation pattern in a lake during summer.

Figure 20 The circulation pattern in a frozen lake and unfrozen lake during winter.

Figure 21 Vrtical distribution along a cross-sectin (Ado River mouth – Maibara section) of water temperature, comparing the results in January and March (2002) calculated by Biwa 3D model.

3.2 Spring Habitat Assessment

The classification and evaluation of spring habitats are necessary to provide basic information, which can be used to establish spring management and biodiversity conservation priorities. However, classifying spring habitats is a challenge since springs are highly variable habitats (Kubikova et al., 2012). Investigators have classified spring habitats based on the geologic origin, rock type, physical properties, chemical composition, discharge rates, groundwater flow path, habitat condition, etc. The new classification of spring types proposed in Chapter 2, focuses on the locations of springs in relation to the main channel of rivers based on differences in water levels. The classifications stress demonstrates the importance of different types of spring habitats associated with the dynamics of flood-plain elements for structuring the spring-associated benthic assemblages.

Various studies in spring ecology have been conducted to identify the influence of springs with different habitat conditions on the invertebrate communities. Myers and Resh (2002) researched the species composition, richness and distribution of macroinvertebrates in springs with different physical conditions and they found that the cold water temperatures were the factors most responsible for species richness of Trichoptera. Wood and Gunn (2003) investigated the physical habitat and discharge patterns of karst (limestone) springbrook systems and their macroinvertebrate fauna to evaluate the roles of habitat structure and flow permanence on macroinvertebrate community composition. They found flow permanence and water temperature exerted more impacts on the

abundance of macroinvertebrates than habitat structure. Kubikova et al. (2012) analyzed the composition of spring fauna in relation to the mesoscale habitat conditions in a geologically homogeneous area and demonstrated the importance of mesoscale differences in spring habitat conditions for benthic assemblages.

The differences in habitat conditions such as stream bed (Wallace and Webster,1996), land use (Resh et al., 1988), substrate type (Buss et al., 2004), water temperature(Townsend et al., 1997; Merritt et al., 2008), habitat stability (Death and Winterbourn, 1995) were pointed out as important environmental factors for structuring the invertebrate communities. Springs and their fauna are also associated with the river and the dynamics of floodplain elements (Scarsbrook et al., 2007). Classifying spring types with the consideration of floodplain elements is important for understanding the impacts of surface water on spring ecosystems. The information can be used to establish spring management and conservation priorities in the river ecosystems.

3.3 Environmental Education Project

Nowadays, more and more scientific research outcomes and information have been provided to solve environmental issues and many of them have achieved great success. As researchers, we promote the ecosystem management practice by providing such knowledge and suggestions to decision-makers who can then set up the management priorities.

During my oversea internship in the European Regional Centre for Ecohydrology (ERCE), I have learned knowledge and bio-technologies of water pollution control projects in Poland. Project EKO- ROB aims to reduce diffuse pollution in the Pilica basin and improve the water quality of Sulejów Reservoir. Project EH-REK is conducted to improve water quality and allow continued recreational use of rehabilitated reservoirs in the city of Łodź. Both projects have developed innovative biotechnologies to achieve the sustainable goal of watershed management. Many traditional environmental strategies for solving water quality problems are costly and less effective due to some constraints. Project EKO-ROB constructed diversified ecotones using vegetations to reduce the diffuse pollution in Pilica basin. The ecohydrological approach used in Project EKO-ROB is integrated strategies for restoring aquatic ecosystems and improving water quality. This cost-effective ecohydrological approach is needed for sustainable water resource management. During the stay at the institution, I also had opportunities to attend some environmental conservation-related field trip and field researches which mainly focused on chemical pollutants and bacterial studies. Participating in

these activities gave me different experiences and knowledge of environmental conservation work. It is also a good chance to have communications with local researchers and local people, gaining a better understanding of local problems.

As long as we are confident about these scientific technologies, we are still facing the challenges of water pollution in the current situation. A part of the pollution comes from local agriculture activities. Ecological buffer zoom has been constructed between agricultural lands and rivers to reduce the pollution, however, due to the technological limits and the lack of understanding from local farmers, the pollution problem is not completely solved. As our understanding of nature and approaches to protect the environment have come sophisticated and complex, the gap between science and public understanding is also becoming a serious issue.

We consider it is essential to not only publish the studies in an academic journal but also share the knowledge and information to the general public through an environmental education project. Publishing the views in an international academic journal is generally a medium for the dissemination of knowledge through the research articles which are more appreciated by the governments and scholars. On the other hand, the activities of environmental education projects provide chances for citizens to learn from researchers and to be exposed to views they might never hear about in their daily life. That then leads to an improved understanding of the environment. From environmental

education activities, people can get a taste of real expertise and insight. The improved environmental awareness, in turn, leads to more environmental commitment and actions as well as more contributions to the decision-making process in environmental management practice.

(1) Project Overview

Project Name: Environmental education on foreign tourists in Japan

Project Type: Community education

Time Period: July ~ October, 2019

Project objectives:

The project aims to develop an environmental education model that can expand the environmental information and scientific knowledge to society and citizens in a way that allows them to make their choices based on their interests and values.

The project focuses on expanding the scientific knowledge of environmental conservation to international tourists in Japan. We designed the environmental education model using the cultural and natural resources of Japanese shrines and tea ceremonies. From this project, tourists can learn Japanese traditional views of nature from Japanese culture, shrines and tea ceremony. We will also

discuss environmental issues such as algae eutrophication and their relationship with human beings, using the water ponds inside the shrine as teaching examples.

We believe that after knowing the important relationship between nature and humans through the process of "知" (Knowing), people can be more aware of the trade-off mechanism between short-term personal gain and long-term communal value and be able to choose which path they wish to take. The project model was designed and discussed among project members and developed through the process of implementation.

The implementation of this project is considered as our process of "思" (Thinking) and "行" (Action). Consequently, we hope our efforts could contribute to public awareness improvement which can lead to the "思" (Thinking) and "行" (Action) of the society.

(2) Project Planning

● Target Group

The targets are foreign tourists who travel to Japan. According to Japan National Tourism Organization (JNTO), the estimated number of international travelers to Japan in January 2019 was about 2.7 million (+7.5% from the previous year, +180,000 travelers), recording the highest figure for 68

January. Foreign tourists have certainly become an important part of Japanese society. The tourist boom is spreading the economic benefits to Japan. It also brings us a great opportunity to communicate and learn.

Tourists were invited to participate in the activities of the project, where we can have the opportunity to communicate, conducting our environmental education and interview. We designed the environmental education model intending to develop the necessary multilateral disciplines to give more influence on people.

● Project Design

We designed an experience activity for international tourists where we can have the opportunities to conduct scientific communication and environmental education. Based on the needs of our target group, we planned to approach the environmental education project by introducing Japanese traditional views of nature from Japanese culture, shrines and tea ceremony, discussing the environmental issues such as algae eutrophication using the water ponds inside the shrine as teaching examples (Figure 22). The ideas of designing the project are shared and discussed among project members and guests. We investigated the methods, tools, and techniques we could use to achieve our goal and objective.

• Brainstorm ideas on necessary multilateral disciplines and possible methods to achieve the purpose of environmental education on foreign tourists.

• Design the contents and schedule of environmental education projects and research necessary information about Sumiyoshi shrine and Tea ceremony.

• Be aware of the characteristics, strengths, and weaknesses of the project. Keep in mind who our target group is and what is the most appropriate, effective and efficient method of achieving our objectives with this target group.

There are different types of environmental education activities and program which have been conducted in Japan so far. I have participated in environmental education activities in Kyoto (Kamo River, Takano River, and Midorogaike) and Shizuoka (Kakida River). These activities educated students and citizens by sampling and identifying benthic invertebrates from rivers, ponds, and springs. In this environmental education project, we are concerned primarily with influencing knowledge and understanding of nature and environment, not only by scientific knowledge but also by traditional views and values of nature. Therefore, we agreed to firstly introduce Japanese traditional shrines and tea ceremony which foreign tourists may have strong interests in. Through the process, we will introduce the issue of algae eutrophication happening in the water ponds of the shrine. The introduction then leads to the relationship between water issues and human beings. We will use both the cultural and natural resources of shrines to achieve our goal of environmental

education.

(3) Project Implementation

We introduce, promote and set the tone for the project through social media (SNS), websites and friend's introduction systems. At the beginning of the project, we opened our schedule as more as possible to draw attention and gain sufficient participants. We hold the project activities in the morning from 9:30 to 12:00 and from 13:00 to 16:00. The group size is set up to 10 people. People are different and will learn differently, therefore we consider to hold each activity on a small-scale to increase communication and effectiveness. The activity duration is set up to 2.5~3 hours, in which we will spend 1.5 hours to introduce the shrines and cultures and 45 minutes to experience the tea ceremony.

Among thousands of shrines in Japan, we chose Sumiyoshi Taisha for several reasons. Sumiyoshi Taisha has almost 2000 years of history. Long history, an important role in Japanese culture and unique architectural styles constructed using natural resources is considered to be the appropriate place to introduce traditional Japanese nature views as well as the values and cultural services that nature has brought to us. Furthermore, there are algae-filled ponds inside the shrine where water turns to green in the cause of algae blooming. These ponds are habitats for wildlife such as fishes, ducks, and turtles. It is a common issue that people appear to feed the wildlife for fun, using bread, rolls, chips and other human "snack food". However, what people don't know is the act of feeding can promote excessive algae growth, leading to decrease oxygen levels, green and smelling water. Moreover, feeding human food to wildlife is also not healthy for the wild animals. I have encountered such an issue during my Poland internship, so I decided to use the ponds as a teaching example for tourists.

After the walk in the shrine, there is a short break and after that, the guests will have a chance to experience a Japanese traditional tea ceremony. During the tea ceremony, we introduce the spirit of tea, the culture of ikebana and the relationships between tea ceremony and nature. We hope the cup of tea can bring guests a moment to experience the engagement with nature, being close to nature and integrate with nature.

We consider increasing people's understanding of nature by approaching a variety of methods. Combined with tea ceremony and Japanese shrines, this environmental education becomes a project that not only uses "information giving" methods but also allows people to experience and participate in.

• Project type: Community education

• Time period: July ~ October

• Activity duration: 2.5 ~ 3 hours

• Group size: Up to 10 people

• Hosted in: English, Chinese (Simplified & Traditional)

• Activity location: Osaka

Figure 22 The content of the environmental education project (Photo by Sun, 2019).

(4) Project Accomplishments

• Project period: 7/1~9/30

• Implementation times (7/1~9/30): 28

• The number of participants in total: 82

• Participant's hometown (country/region): United States, Canada, China (Mainland), • Hong Kong, Taiwan, Mexico.

• Language: English, Chinese

The project has been conducted for more than three months since July. In total, we have implemented 28 times of activities (7 times per month) either in English or Chinese. By the end of September, 82 foreign tourists participated in our project (Figure 23). The guests are from the United States, Canada, China (Mainland), Hong Kong, Taiwan, and Mexico.

Figure 23 The general information (hometown) of the participants.

The purpose of the project is to introduce Japanese traditional cultures and values on nature and educate people on scientific knowledge about our environment and the influence of human activities. The project aims to develop an environmental education model for foreign tourists who come to visit Japan. Therefore, we focused more on how to develop and improve our project. We have improved our project content during the implementation and promoted it to more people. We hope this project

will be of great benefit for people interested or involved in planning and running community education / environmental education projects. It can be used by:

• Individuals and groups

• Community action group

• Environment conservation groups

• River management committees

• Schools and universities

• Government and non-government agencies

Environmental education is important for environmental conservation and raising awareness. However, it is not the only factor that influences people's thinking and actions toward nature. The project may not result in an immediate social change or public awareness improvement in the short term. The environmental education model designed and developed in this project should be considered as one part of contributions that address the issues which in turn leads to people's arising awareness and actions.

Conclusions and Future Directions

The special characteristics of springs, the locations of springs at the interface between surface water and groundwater ecosystems, have led to a high value of biodiversity and high contribution to ecosystem services. Human source use and management can affect the conservation of groundwater and springs spring habitat and biodiversity. Given the susceptibility of springs and groundwaters to modification from land-use change and water abstraction, many species should be considered at risk, and urgent attention is required to incorporate biological indicators into assessment of groundwater and spring habitat classification into conservation and management of the freshwater system. Environmental education is another important factor that should be considered during scientific communication and the decision-making process.

We expected more attention to spring habitats and their dependent species. It becomes imperative to explore this issue considering the potential risks of temperature variation threatened by climate change. More efforts should be made on genomic approaches, cold hardiness, and thermotolerance studies to provide us additional information on stenothermal species. Exploring as broad a range of issues as possible also need cross-study approaches. More integrative studies that combine hydrogeology, engineering, ecology, education and philosophy are needed to achieve sustainable river management. The benefits should be readily accessible to the managers, policymakers and public.

Acknowledgments

I would like to express my gratitude to all those who helped me during the writing of this thesis. My deepest gratitude goes first and foremost to Professor Yosuke YAMASHIKI and Professor Yasuhiro TAKEMON my supervisors, Professor Liang Zhao my mentor, for their constant encouragement and guidance. I am also deeply indebted to all the other tutors and teachers in the Graduate school of Advanced Integrated Studies in Human Survivability for their direct and indirect help to me. Special thanks should go to the Leading Program in Graduate school of Advanced Integrated Studies and the Hodaka Sedimentation Observatory, Kyoto University for the support and assistance in the field, and all those who provided who have helped in carrying out the research.

Chapter 1 is accepted:

Ye SUN, Yasuhiro TAKEMON, Yosuke YAMASHIKI. (in press) Freshwater spring indicator taxa of benthic invertebrates. Ecohydrology & Hydrobiology, 2019.

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Appendix 1 Invertebrate indicators of freshwater springs (Types of spring dependece, “G”: Groundwater dependent species, “C”: coldwater stenothermal organisms, "O": other possible indicator species).

References No.1 - No.29 (Table 1) Type Class Order Family Taxa 1 1 2 3 4 5 6 7 8 9 10 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 1

1 Turbelaria Tricladida Dugesiidae Neppia montana C 2 Turbelaria Tricladida Planariidae Crenobia alpina C + + + + 3 Turbelaria Tricladida Planariidae Dugesia japonica C + + 4 Turbelaria Tricladida Planariidae Dugesia dorotocephala C 5 Turbelaria Tricladida Planariidae Dugesia gonocephala C + 6 Turbelaria Tricladida Planariidae Dugesia lugubris C + 7 Turbelaria Tricladida Planariidae Dugesia spp. C + +

84 8 Turbelaria Tricladida Planariidae Polycelis felina C + +

9 Turbelaria Tricladida Planariidae Polycelis nigra. C + 10 Turbelaria Tricladida Planariidae Polycelis tibetica C + 11 Turbelaria Tricladida Planariidae Polycelis spp. C + + + 12 Turbelaria Tricladida Planariidae Phagocata morgani C 13 Turbelaria Tricladida Planariidae Phagocata sp. C 14 Bivalvia Pelecypoda Sphaeriidae Sphaerium corneum C 15 Gastropoda Heterostropha Valvatidae Valvata piscinalis G + 16 Gastropoda Littorinimorpha Hydrobiidae Belgrandiella kuesteri G + 17 Gastropoda Littorinimorpha Hydrobiidae Belgrandiella fontinalis G + 18 Gastropoda Littorinimorpha Hydrobiidae Bythinella bavarica G 19 Gastropoda Littorinimorpha Hydrobiidae Bythinella cylindrica G 20 Gastropoda Littorinimorpha Hydrobiidae Bythinella opaca opaca G

Appendix 1 (continued). References No.1 - No.29 (Table 1) Type Class Order Family Taxa 1 1 2 3 4 5 6 7 8 9 10 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 1

21 Gastropoda Littorinimorpha Hydrobiidae Bythinella schmidti G + 22 Gastropoda Littorinimorpha Hydrobiidae Bythinella spp. G + + + 23 Gastropoda Littorinimorpha Hydrobiidae Bythinella dnkeri G + 24 Gastropoda Littorinimorpha Hydrobiidae Fontigens nickliniana G 25 Gastropoda Littorinimorpha Hydrobiidae Graziana sp. G + 26 Gastropoda Littorinimorpha Hydrobiidae Iglica hauffeni G + 27 Gastropoda Littorinimorpha Hydrobiidae Potamopyrgus doci G 28 Gastropoda Littorinimorpha Hydrobiidae Sadleriana supercarinata G + 29 Clitellata Enchytraeida Enchytraeidae Marionina argentea G +

30 Clitellata Enchytraeida Enchytraeidae Marionina sambugarae G Arhynchobdelli 31 Hirudinea da Erpobdellidae Erpobdella octoculata C + + + 32 Oligochaeta Lumbriculida Lumbriculidae Lumbriculus variegatus O 33 Oligochaeta Tubificida Naididae Aberrantidrilus stephaniae G Ilyodrilus frantzi 34 Oligochaeta Tubificida Naididae Brinkhurst O 35 Oligochaeta Tubificida Naididae Nais variabilis O + 36 Oligochaeta Tubificida Naididae Opidonais serpentina O Psammoryctides 37 Oligochaeta Tubificida Naididae deserticola O 38 Oligochaeta Tubificida Naididae Quistadrilus multisetosus O Trombidiforme 39 Arachnida s Hygrobatidae Atractides panniculatus O + +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Type Class Order Family Taxa 1 1 2 3 4 5 6 7 8 9 10 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 1

40 Arachnida Trombidiformes Hygrobatidae Sperchon squamosus O

41 Arachnida Araneae Oribatida Oribatida sp. C 42 Ostracoda Podocopida Candonidae Candona acuminata G 43 Ostracoda Podocopida Candonidae Candona lindneri G + 44 Ostracoda Podocopida Candonidae Candona hyalina G 45 Ostracoda Podocopida Candonidae Candona gr. neglecta G + 46 Ostracoda Podocopida Candonidae Cryptocandona vavrai G + 47 Ostracoda Podocopida Candonidae Fabaeformiscandona sp. G +

86 48 Ostracoda Podocopida Cyprididae Ilyocypris bradyi G +

49 Ostracoda Podocopida Cyprididae Ilyocypris gibba G 50 Ostracoda Podocopida Cyprididae Ilyocypris biplicata G 51 Ostracoda Podocopida Cyprididae Ilyocypris beauchampi G 52 Ostracoda Podocopida Cyprididae Cyprinotus salinus G 53 Ostracoda Podocopida Cyprididae Cyclocypris brevisetosa G 54 Ostracoda Podocopida Cyprididae Cypris pubera G 55 Ostracoda Podocopida Cyprididae Chydorus sphaericus G 56 Ostracoda Podocopida Cyprididae Chydorus sp. G 57 Ostracoda Podocopida Cyprididae Cyprididae gen. spp. G + 58 Ostracoda Podocopida Cyprididae Eucyclops serrulatus G 59 Ostracoda Podocopida Cyprididae Eucypris inflata G 60 Ostracoda Podocopida Cyprididae Iliocypris inermis G

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

61 Ostracoda Podocopida Cyprididae Potamocypris pallida G + 62 Ostracoda Podocopida Cyprididae Psychrodromus betharrami G + 63 Ostracoda Podocopida Cyprididae Psychrodromus fontinalis G + + 64 Ostracoda Podocopida Cyprididae Psychrodromus olivaceus G + 65 Ostracoda Podocopida Cyprididae Scottia pseudobrowniana G + 66 Ostracoda Podocopida Cyprididae Stenocypris subterranea G 67 Ostracoda Podocopida Cyprididae Cavernocypris subterranea G + Pseudolimnocythere 68 Ostracoda Podocopida Loxoconchidae hypogea G + + + + +

87 69 Ostracoda Ostracoda Ostracoda Ostracoda gen. spp. G

70 Ostracoda Podocopida Cyprididae Heterocypris oblonga G 71 Ostracoda Podocopida Cyprididae Herpetocypris chevreuxi G 72 Ostracoda Podocopida Cyprididae Humphcypris subterranea G 73 Ostracoda Podocopida Cyprididae Humphcypris anomala G 74 Ostracoda Podocopida Cyprididae Humphcypris brevisetosa G 75 Ostracoda Podocopida Cyprididae Humphcypris chappuisi G 76 Ostracoda Podocopida Cyprididae Humphcypris decipiens G 77 Ostracoda Podocopida Cyprididae Humphcypris exigua G 78 Ostracoda Podocopida Cyprididae Humphcypris greenwoddi G 79 Ostracoda Podocopida Cyprididae Humphcypris leleupi G 80 Ostracoda Podocopida Cyprididae Humphcypris sewelli G

Appendix 1 (continued). References No.1 - No.29 (Table 1) Type Class Order Family Taxa 1 1 1 2 3 4 5 6 7 8 9 10 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 1 2

81 Ostracoda Podocopida Cyprididae Humphcypris thysvillensis G 82 Ostracoda Podocopida Cyprididae Humphcypris tumens G 83 Maxillopoda Cyclopoida Cyclopidae Acanthocyclops agamus G + 84 Maxillopoda Cyclopoida Cyclopidae Acanthocyclops sp. G + 85 Maxillopoda Cyclopoida Cyclopidae Diacyclops bicuspidatus G 86 Maxillopoda Cyclopoida Cyclopidae Diacyclops copepodids G + 87 Maxillopoda Cyclopoida Cyclopidae Diacyclops disjunctus G + 88 Maxillopoda Cyclopoida Cyclopidae Diacyclops languidoides G + 89 Maxillopoda Cyclopoida Cyclopidae Eucyclops serrulatus G

90 Maxillopoda Cyclopoida Cyclopidae Eucyclops sp. G + 91 Maxillopoda Cyclopoida Cyclopidae Megacyclops viridis G + 92 Maxillopoda Cyclopoida Cyclopidae Macrocyclops albidus G 93 Maxillopoda Cyclopoida Cyclopidae Macrocyclops sp. G 94 Maxillopoda Cyclopoida Cyclopidae Itocyclops sp. G + 95 Maxillopoda Cyclopoida Cyclopidae Speocyclops kieferi G + 96 Maxillopoda Cyclopoida Cyclopidae speocyclops racovitzai G + 97 Maxillopoda Cyclopoida Cyclopidae Paracyclops fimbriatus G 98 Maxillopoda Cyclopoida Cyclopidae Paracyclops imminutus G + 99 Maxillopoda Cyclopoida Cyclopidae Afrocyclops sp. G Harpacticoid- Canthocampti- Attassaheyella 100 Maxillopoda a dae (Attheyella) crassa G +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 Atheyella 101 Maxillopoda Harpacticoida Canthocamptidae nordenskioldii C Bryocamptus (Rheocamptus) 102 Maxillopoda Harpacticoida Canthocamptidae copepodites G +

Bryocamptus 103 Maxillopoda Harpacticoida Canthocamptidae cuspidatus G + 104 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus dacicus G 105 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus nivalis G

106 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus rhaeticus G + +

Bryocamptus 107 Maxillopoda Harpacticoida Canthocamptidae vandouwei G +

108 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus pygmaeus G +

109 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus ethinatus G +

110 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus tatrensis G +

111 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus typhlops G +

112 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus zschokkei G +

113 Maxillopoda Harpacticoida Canthocamptidae Cypria ophthalmica G

Appendix 1 (continued).

Type References No.1 - No.29 (Table 1) Class Order Family Taxa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

Elaphoidella 114 Maxillopoda Harpacticoida Canthocamptidae pseudophreatica G + Elaphoidella 115 Maxillopoda Harpacticoida Canthocamptidae phreatica G + Epactophanes 116 Maxillopoda Harpacticoida Canthocamptidae richardi G + 117 Maxillopoda Harpacticoida Canthocamptidae Hypocamptus brehmi G + Moraria (Moraria) 118 Maxillopoda Harpacticoida Canthocamptidae alpina G + 119 Maxillopoda Harpacticoida Canthocamptidae Moraria poppei G +

120 Maxillopoda Harpacticoida Canthocamptidae Moraria stankovitchi G + 121 Maxillopoda Harpacticoida Canthocamptidae Moraria varica G +

122 Maxillopoda Harpacticoida Canthocamptidae Stygepactophanes sp. G + 123 Maxillopoda Harpacticoida Parastenocarididae Parastenocaris sp. G + + Harpacticoida gen. 124 Maxillopoda Harpacticoida Harpacticoida spp. G 125 Malacostraca Amphipoda Seborgiidae Seborgia relicta G Jesogammarus 126 Malacostraca Amphipoda Anisogammaridae fluvialis C + Crangonyx 127 Malacostraca Amphipoda Crangonyctidae pseudogracilis G + Crangonyx 128 Malacostraca Amphipoda Crangonyctidae subterraneus G + 129 Malacostraca Amphipoda Crangonyctidae Crangonyx floridanus G + +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

130 Malacostraca Amphipoda Crangonyctidae Synurella ambulans O + 131 Malacostraca Amphipoda Crangonyctidae Stygobromus cf. ambulans G + 132 Malacostraca Amphipoda Crangonyctidae Stygobromus pecki G 133 Malacostraca Amphipoda Crangonyctidae Stygobromus russelli G 134 Malacostraca Amphipoda Crangonyctidae Stygobromus gen. spp. G 135 Malacostraca Amphipoda Gammaridae Echinogammarus gen. spp. G + + 136 Malacostraca Amphipoda Gammaridae Gammarus balcanicus G + 137 Malacostraca Amphipoda Gammaridae Gammarus fossarum G + + + + 138 Malacostraca Amphipoda Gammaridae Gammarus lacustris G

91 139 Malacostraca Amphipoda Gammaridae Gammarus pulex G + + +

140 Malacostraca Amphipoda Gammaridae Gammarus minus G 141 Malacostraca Amphipoda Hadziidae Mexiweckelia hardeni G 142 Malacostraca Amphipoda Hyalellidae Hyalella azteca G Mesogammari- 143 Malacostraca Amphipoda dae Eoniphargus kojimai G + + 144 Malacostraca Amphipoda Niphargidae Niphargus aquilex G + 145 Malacostraca Amphipoda Niphargidae Niphargus fontanus G + 146 Malacostraca Amphipoda Niphargidae Niphargus foreli G + 147 Malacostraca Amphipoda Niphargidae Niphargus laisi G + 148 Malacostraca Amphipoda Niphargidae Niphargus sp. aff. puteanus G + 149 Malacostraca Amphipoda Niphargidae Niphargus sp. G + + + + 150 Malacostraca Amphipoda Niphargidae Niphargus gineti G +

Appendix 1 (continued). References No.1-29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

151 Malacostraca Amphipoda Niphargidae Niphargus kochianus G + 152 Malacostraca Amphipoda Niphargidae Niphargus longidactylus G + 153 Malacostraca Amphipoda Niphargidae Niphargus gr. stygius G + 154 Malacostraca Amphipoda Niphargidae Niphargus tatrensis G + Paraleptamph- 155 Malacostraca Amphipoda opidae Paraleptamphopus sp. G Pseudocrango- 156 Malacostraca Amphipoda nyctidae Pseudocrangonyx sp. G + + 157 Malacostraca Amphipoda Pontogeneiidae Sternomoera sp. G + 158 Malacostraca Decapoda Cambaridae Orconectes immunis G

92 159 Malacostraca Decapoda Potamidae Geothelphusa dehaani C +

160 Malacostraca Isopoda Asellidae Proasellus deminutus O + 161 Insecta Ephemeroptera Baetidae Baetis alpinus C + + + 162 Insecta Ephemeroptera Baetidae Baetis vernus C + 163 Insecta Ephemeroptera Baetidae Baetis vagans C Maccaffertium 164 Insecta Ephemeroptera Heptageniidae meririvulanum C 165 Insecta Ephemeroptera Heptageniidae Rhithrogena loyolea C + 166 Insecta Odonata Calopterygidae Argia vivida C 167 Insecta Odonata Calopterygidae Calopteryx atrata C + + 168 Insecta Odonata Calopterygidae Calopteryx sp. C 169 Insecta Plecoptera Capniidae Capnopsis schilleri C + 170 Insecta Plecoptera Chloroperlidae Alloperla mediana C

Appendix 1 (continued). References No.1-29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

171 Insecta Plecoptera Chloroperlidae Alloperla sp. C 172 Insecta Plecoptera Chloroperlidae Siphonoperla burmeisteri C + 173 Insecta Plecoptera Chloroperlidae Siphonoperla montana C + 174 Insecta Plecoptera Leuctridae Leuctra braueri C + + 175 Insecta Plecoptera Leuctridae Leuctra digitata C + 176 Insecta Plecoptera Leuctridae Leuctra tenella C 177 Insecta Plecoptera Leuctridae Leuctra rosinae C + 178 Insecta Plecoptera Leuctridae Perlomyia sp. C 179 Insecta Plecoptera Leuctridae Leuctra juv. [group A] C +

93 180 Insecta Plecoptera Leuctridae Leuctra juv. [group B] C +

181 Insecta Plecoptera Nemouridae Amphinemura standfussi C + + 182 Insecta Plecoptera Nemouridae Amphinemura sulcicollis C + 183 Insecta Plecoptera Nemouridae Protonemura meyeri C + 184 Insecta Plecoptera Nemouridae Protonemura auberti C + + 185 Insecta Plecoptera Nemouridae Protonemura brevistyla C + 186 Insecta Plecoptera Nemouridae Protonemura lateralis C + 187 Insecta Plecoptera Nemouridae Protonemura nimborum C + 188 Insecta Plecoptera Nemouridae Protonemura sp. C + + 189 Insecta Plecoptera Nemouridae Nemoura fulva C + 190 Insecta Plecoptera Nemouridae Nemoura flexuosa C +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

191 Insecta Plecoptera Nemouridae Nemoura albidipennis C 192 Insecta Plecoptera Nemouridae Nemoura cinerea C + + + + + + 193 Insecta Plecoptera Nemouridae Nemoura completa C 194 Insecta Plecoptera Nemouridae Nemoura obtusa C + 195 Insecta Plecoptera Nemouridae Nemoura picteii C + + + 196 Insecta Plecoptera Nemouridae Nemoura mortoni C + 197 Insecta Plecoptera Nemouridae Nemoura sinuata C + 198 Insecta Plecoptera Nemouridae Nemoura vallicularia C 199 Insecta Plecoptera Nemouridae Nemoura gen. spp. C + + +

94 200 Insecta Plecoptera Nemouridae Nemurella picteti C + + + + + + +

201 Insecta Plecoptera Notonemouridae Malenka flexura C 202 Insecta Plecoptera Notonemouridae Spaniocerca zelandica C 203 Insecta Plecoptera Notonemouridae Spaniocerca sp. C 204 Insecta Plecoptera Notonemouridae Cristaperla sp. C 205 Insecta Plecoptera Notonemouridae Halticoperla viridans C 206 Insecta Plecoptera Perlodidae Dictyogenus alpinum C + 207 Insecta Plecoptera Perlodidae Dictyogenus fontium C + 208 Insecta Plecoptera Perlodidae Diura nanseni C + 209 Insecta Plecoptera Perlodidae Isogenus subvarians C 210 Insecta Plecoptera Perlidae Acroneuria abnormis C

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

211 Insecta Plecoptera Perlidae Eccoptura xanthenes C 212 Insecta Plecoptera Perlidae Kamimuria quadrata C + 213 Insecta Plecoptera Perlidae Isoperla albanica C + 214 Insecta Plecoptera Perlidae Isoperla rivulorum C + 215 Insecta Plecoptera Perlidae Perta sp. C + 216 Insecta Plecoptera Peltoperlidae Peltoperla arcuata C 217 Insecta Plecoptera Gripopterygidae Megaleptoperla grandis C 218 Insecta Plecoptera Gripopterygidae Megaleptoperla sp. C 219 Insecta Plecoptera Gripopterygidae Taraperla howsei C

95 220 Insecta Plecoptera Gripopterygidae Zelandobius unicolor C

221 Insecta Plecoptera Gripopterygidae Zelandobius pilosus C 222 Insecta Plecoptera Gripopterygidae Zelandobius confusus C 223 Insecta Plecoptera Gripopterygidae Zelandobius furcillatus C 224 Insecta Plecoptera Gripopterygidae Zelandobius illiesi C 225 Insecta Plecoptera Gripopterygidae Zelandobius sp. C 226 Insecta Plecoptera Gripopterygidae Zelandoperla decorata C 227 Insecta Plecoptera Gripopterygidae Zelandoperla sp. C 228 Insecta Plecoptera Eustheniidae Stenoperla maclellani C 229 Insecta Megaloptera Sialidae Sialis fuliginosa C + + 230 Insecta Megaloptera Corydalidae Archichauliodes diversus C

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

231 Insecta Megaloptera Corydalidae Parachauliodes japonicus C + 232 Insecta Megaloptera Nannochoristidae Nannochorista philpotti C 233 Insecta Megaloptera Nannochoristidae Nannochorista sp. C 234 Insecta Neuroptera Osmylidae Kempynus sp. C 235 Insecta Neuroptera Osmylidae Osmylidae gen. sp. G 236 Insecta Trichoptera Apataniidae Apatania eatoniana O + 237 Insecta Trichoptera Brachycentridae Brachycentridae gen. spp. O + + 238 Insecta Trichoptera Hydrobiosidae Psyllobetina attunga C 239 Insecta Trichoptera Hydroptilidae Orthotrichia gen. spp. O

96 240 Insecta Trichoptera Philopotamidae Dolophilodes sp. C +

241 Insecta Trichoptera Philopotamidae Wormaldia copiosa C + 242 Insecta Trichoptera Philopotamidae Wormaldia occipitalis C + 243 Insecta Trichoptera Philopotamidae Wormaldia sp. C + + + 244 Insecta Trichoptera Phryganeidae Ptilostomis sp. C 245 Insecta Trichoptera Polycentropodidae Plectrocnemia conspersa C + + + + + + + + 246 Insecta Trichoptera Polycentropodidae Plectrocnemia geniculata C + + + + 247 Insecta Trichoptera Polycentropodidae Plectrocnemia sp. C + + + + Polycentropus 248 Insecta Trichoptera Polycentropodidae flavomaculatus C + 249 Insecta Trichoptera Polycentropodidae Polycentropus sp. C 250 Insecta Trichoptera Psychomyiidae Tinodes unicolor C +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

251 Insecta Trichoptera Rhyacophilidae Rhyacophila towadensis C 252 Insecta Trichoptera Rhyacophilidae Rhyacophila sp.X-2 C + + + 253 Insecta Trichoptera Rhyacophilidae Rhyacophila sp.RD C + + + 254 Insecta Trichoptera Rhyacophilidae Rhyacophila fasciata Hagen C + 255 Insecta Trichoptera Rhyacophilidae Rhyacophila shikotsuensis C 256 Insecta Trichoptera Rhyacophilidae Rhyacophila lobifera C 257 Insecta Trichoptera Rhyacophilidae Rhyacophila lezeyi C + 258 Insecta Trichoptera Rhyacophilidae Rhyacophila impar C 259 Insecta Trichoptera Rhyacophilidae Rhyacophila transquila C

97 260 Insecta Trichoptera Rhyacophilidae Rhyacophila brevicephara C +

261 Insecta Trichoptera Rhyacophilidae Rhyacophila brunnea C 262 Insecta Trichoptera Rhyacophilidae Rhyacophila dorsalis C + + 263 Insecta Trichoptera Rhyacophilidae Rhyacophila nigra C + 264 Insecta Trichoptera Rhyacophilidae Rhyacophila spp. C + + + 265 Insecta Trichoptera Sericostomatidae Sericostoma personatum C + + + 266 Insecta Trichoptera Sericostomatidae Sericostoma sp. C + + 267 Insecta Lepidoptera Pyralidae Nymphulinae gen.spp. C + 268 Insecta Diptera Athericidae Atherix sp. C + + + 269 Insecta Diptera Chironomidae Diamesa bertrami C + 270 Insecta Diptera Chironomidae Diamesa cinerella C +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

271 Insecta Diptera Chironomidae Diamesa latitarsis C + 272 Insecta Diptera Chironomidae Diamesa gen. spp. C + 273 Insecta Diptera Chironomidae Anzacladius sp. C 274 Insecta Diptera Chironomidae Boreochlus sp. C 275 Insecta Diptera Chironomidae Metriocnemus fuscipes C + 276 Insecta Diptera Chironomidae Parochlus sp. C 277 Insecta Diptera Chironomidae Prodiamesa bathyphila C 278 Insecta Diptera Chironomidae Prodiamesa olivacea C + + + 279 Insecta Diptera Chironomidae Prodiamesa sp. C

98 280 Insecta Diptera Chironomidae Rheocricotopus atripes C + +

281 Insecta Diptera Chironomidae Rheocricotopus effusus C + + 282 Insecta Diptera Chironomidae Rheocricotopus fuscipes C + 283 Insecta Diptera Chironomidae Stempellinella flavidula C + 284 Insecta Diptera Chironomidae Stilocladius montanus C + 285 Insecta Diptera Chironomidae Boreoheptagyia legeri C + 286 Insecta Diptera Chironomidae Boreoheptagyia sp. C 287 Insecta Diptera Chironomidae Pagastia sp.1 C + + 288 Insecta Diptera Chironomidae Pagastia sp.2 C + + + 289 Insecta Diptera Dixidae Dixa (Dixella) californica C 290 Insecta Diptera Dixidae Dixa dilatata C + +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

291 Insecta Diptera Dixidae Dixa inextricata C 292 Insecta Diptera Dixidae Dixa marculata -agg. C + + 293 Insecta Diptera Dixidae Dixa modesta C 294 Insecta Diptera Dixidae Dixa naevis C 295 Insecta Diptera Dixidae Dixa nipponica C + 296 Insecta Diptera Dixidae Dixa neoaliciae C 297 Insecta Diptera Dixidae Dixa submaculata C + 298 Insecta Diptera Dixidae Dixa similis C 299 Insecta Diptera Dixidae Dixa sp. C + + + + +

99 300 Insecta Diptera Dixidae Paradixa fuscinervis C

301 Insecta Diptera Dixidae Paradixa harrisi C 302 Insecta Diptera Dixidae Paradixa neozelandica C 303 Insecta Diptera Dixidae Dixidae gen.spp. C + + + + + 304 Insecta Diptera Dolichopodidae Dolichopodidae n. det. C 305 Insecta Diptera Psychodidae Pericoma albitarsis C 306 Insecta Diptera Psychodidae Pericoma bipunctata C 307 Insecta Diptera Psychodidae Pericoma spp. C + + + 308 Insecta Diptera Psychodidae Psychoda parthenogenitica C 309 Insecta Diptera Psychodidae Psychoda sp. C + Telmatoscopus 310 Insecta Diptera Psychodidae albipunctatus C

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

311 Insecta Diptera Psychodidae Telmatoscopus aldrichanus C 312 Insecta Diptera Psychodidae Telmatoscopus basalis C 313 Insecta Diptera Psychodidae Telmatoscopus bipunctata C Telmatoscopus 314 Insecta Diptera Psychodidae quadripunctatus C 315 Insecta Diptera Psychodidae Thornburghiella slossoni C 316 Insecta Diptera Psychodidae Threticus bicolor C 317 Insecta Diptera Psychodidae Tinearia alternicula C 318 Insecta Diptera Chironomidae Pentaneurella sp. C

100 319 Insecta Diptera Chironomidae Pentaneura sp. C

320 Insecta Diptera Thaumaleidae Thaumalea testacea O + 321 Insecta Diptera Thaumaleidae Thaumaleidae gen. spp. C + 322 Insecta Coleoptera Dytiscidae Agabus biguttatus C + + 323 Insecta Coleoptera Dytiscidae Agabus bipunstulatus C + + + + + 324 Insecta Coleoptera Dytiscidae Agabus didymus C + 325 Insecta Coleoptera Dytiscidae Agabinus glabrellus C 326 Insecta Coleoptera Dytiscidae Agabus guttatus C + + + 327 Insecta Coleoptera Dytiscidae Agabus lapponicus C + 328 Insecta Coleoptera Dytiscidae Agabus melanarius C + + + 329 Insecta Coleoptera Dytiscidae Agabus spp. C + + + + + 330 Insecta Coleoptera Dytiscidae Antiporus stigosulus C

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

331 Insecta Coleoptera Dytiscidae Coelambus sp. C + 332 Insecta Coleoptera Dytiscidae Colymbetes sp. C + 333 Insecta Coleoptera Dytiscidae Colymbetinae gen. spp. C + 334 Insecta Coleoptera Dytiscidae Graptodytes sp. C + 335 Insecta Coleoptera Dytiscidae Haideoporus texanus C 336 Insecta Coleoptera Dytiscidae Hydroporus memnonius C + + 337 Insecta Coleoptera Dytiscidae Hydroporus nigrita C + + + 338 Insecta Coleoptera Dytiscidae Hydroporus obsoletus C + 339 Insecta Coleoptera Dytiscidae Hydroporus palustris C + 101 340 Insecta Coleoptera Dytiscidae Hydroporus incognitus C + + +

341 Insecta Coleoptera Dytiscidae Hydroporus tristis C + 342 Insecta Coleoptera Dytiscidae Hydroporus sp. C + 343 Insecta Coleoptera Dytiscidae Hydroglyphus sp. C 344 Insecta Coleoptera Dytiscidae Huxelhydrus syntheticus C 345 Insecta Coleoptera Dytiscidae Liodessus deflectus C 346 Insecta Coleoptera Dytiscidae Platambus maculatus C + 347 Insecta Coleoptera Dytiscidae Platambus sp. C + 348 Insecta Coleoptera Dytiscidae Rhantus sp. C + Stictotarsus 349 Insecta Coleoptera Dytiscidae duodecimpustulatus C +

Appendix 1 (continued). References No.1 - No.29 (Table 1) Class Order Family Taxa Type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

350 Insecta Coleoptera Hydrophilidae Anacaena lutescens C + + + 351 Insecta Coleoptera Hydrophilidae Anacaena globulus C + + + 352 Insecta Coleoptera Hydrophilidae Aliplus sp. C + 353 Insecta Coleoptera Hydrophilidae Ametor latus C 354 Insecta Coleoptera Hydrophilidae Berosus affinis C + 355 Insecta Coleoptera Hydrophilidae Berosus nigriceps C 356 Insecta Coleoptera Hydrophilidae Berosus sp. C + 357 Insecta Coleoptera Hydrophilidae Coelostoma orbiculare C + 358 Insecta Coleoptera Hydrophilidae Helophorus brevipalpis C + + + + 102 359 Insecta Coleoptera Hydrophilidae Helophorus fuscipes C + 360 Insecta Coleoptera Hydrophilidae Helophorus strigifrons C + 361 Insecta Coleoptera Hydrophilidae Helophorus sp. C + + + Hydrobius fuscipes 362 Insecta Coleoptera Hydrophilidae (Linnaeus) C + + + Hydrobius fuscipes 363 Insecta Coleoptera Hydrophilidae (Fabricius) C + 364 Insecta Coleoptera Hydrophilidae Hydrobius sp. C

Appendix 1 (continued). References No.30 - No.59 (Table 1) Class Order Family Taxa Type 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59

1 Turbelaria Tricladida Dugesiidae Neppia montana C + 2 Turbelaria Tricladida Planariidae Crenobia alpina C + 3 Turbelaria Tricladida Planariidae Dugesia japonica C 4 Turbelaria Tricladida Planariidae Dugesia dorotocephala C + 5 Turbelaria Tricladida Planariidae Dugesia gonocephala C 6 Turbelaria Tricladida Planariidae Dugesia lugubris C 7 Turbelaria Tricladida Planariidae Dugesia spp. C + 8 Turbelaria Tricladida Planariidae Polycelis felina C 9 Turbelaria Tricladida Planariidae Polycelis nigra. C 103 10 Turbelaria Tricladida Planariidae Polycelis tibetica C 11 Turbelaria Tricladida Planariidae Polycelis spp. C 12 Turbelaria Tricladida Planariidae Phagocata morgani C + 13 Turbelaria Tricladida Planariidae Phagocata sp. C + 14 Bivalvia Pelecypoda Sphaeriidae Sphaerium corneum C + 15 Gastropoda Heterostropha Valvatidae Valvata piscinalis G + 16 Gastropoda Littorinimorpha Hydrobiidae Belgrandiella kuesteri G 17 Gastropoda Littorinimorpha Hydrobiidae Belgrandiella fontinalis G 18 Gastropoda Littorinimorpha Hydrobiidae Bythinella bavarica G + 19 Gastropoda Littorinimorpha Hydrobiidae Bythinella cylindrica G + 20 Gastropoda Littorinimorpha Hydrobiidae Bythinella opaca opaca G +

Appendix 1 (continued). References No.30 - No.59 (Table 1) Type Class Order Family Taxa 5 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 9

21 Gastropoda Littorinimorpha Hydrobiidae Bythinella schmidti G 22 Gastropoda Littorinimorpha Hydrobiidae Bythinella spp. G 23 Gastropoda Littorinimorpha Hydrobiidae Bythinella dnkeri G 24 Gastropoda Littorinimorpha Hydrobiidae Fontigens nickliniana G + 25 Gastropoda Littorinimorpha Hydrobiidae Graziana sp. G 26 Gastropoda Littorinimorpha Hydrobiidae Iglica hauffeni G 27 Gastropoda Littorinimorpha Hydrobiidae Potamopyrgus doci G + Sadleriana 28 Gastropoda Littorinimorpha Hydrobiidae supercarinata G 104 29 Clitellata Enchytraeida Enchytraeidae Marionina argentea G + 30 Clitellata Enchytraeida Enchytraeidae Marionina sambugarae G + 31 Hirudinea Arhynchobdellida Erpobdellidae Erpobdella octoculata C + 32 Oligochaeta Lumbriculida Lumbriculidae Lumbriculus variegatus O + Aberrantidrilus 33 Oligochaeta Tubificida Naididae stephaniae G + Ilyodrilus frantzi 34 Oligochaeta Tubificida Naididae Brinkhurst O + 35 Oligochaeta Tubificida Naididae Nais variabilis O + 36 Oligochaeta Tubificida Naididae Opidonais serpentina O + Psammoryctides 37 Oligochaeta Tubificida Naididae deserticola O + 38 Oligochaeta Tubificida Naididae Quistadrilus multisetosus O + 39 Arachnida Trombidiformes Hygrobatidae Atractides panniculatus O 40 Arachnida Trombidiformes Hygrobatidae Sperchon squamosus O +

Appendix 1 (continued). References No.30 - No.59 (Table 1) Type Class Order Family Taxa 5 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 9

41 Arachnida Araneae Oribatida Oribatida sp. C + 42 Ostracoda Podocopida Candonidae Candona acuminata G + 43 Ostracoda Podocopida Candonidae Candona lindneri G + 44 Ostracoda Podocopida Candonidae Candona hyalina G + 45 Ostracoda Podocopida Candonidae Candona gr. neglecta G 46 Ostracoda Podocopida Candonidae Cryptocandona vavrai G 47 Ostracoda Podocopida Candonidae Fabaeformiscandona sp. G 48 Ostracoda Podocopida Cyprididae Ilyocypris bradyi G + 49 Ostracoda Podocopida Cyprididae Ilyocypris gibba G +

105 50 Ostracoda Podocopida Cyprididae Ilyocypris biplicata G +

51 Ostracoda Podocopida Cyprididae Ilyocypris beauchampi G + 52 Ostracoda Podocopida Cyprididae Cyprinotus salinus G + 53 Ostracoda Podocopida Cyprididae Cyclocypris brevisetosa G + 54 Ostracoda Podocopida Cyprididae Cypris pubera G + 55 Ostracoda Podocopida Cyprididae Chydorus sphaericus G + 56 Ostracoda Podocopida Cyprididae Chydorus sp. G + 57 Ostracoda Podocopida Cyprididae Cyprididae gen. spp. G 58 Ostracoda Podocopida Cyprididae Eucyclops serrulatus G + 59 Ostracoda Podocopida Cyprididae Eucypris inflata G + 60 Ostracoda Podocopida Cyprididae Iliocypris inermis G +

Appendix 1 (continued). References No.30 - No.59 (Table 1) Type Class Order Family Taxa 5 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 9

61 Ostracoda Podocopida Cyprididae Potamocypris pallida G 62 Ostracoda Podocopida Cyprididae Psychrodromus betharrami G 63 Ostracoda Podocopida Cyprididae Psychrodromus fontinalis G 64 Ostracoda Podocopida Cyprididae Psychrodromus olivaceus G 65 Ostracoda Podocopida Cyprididae Scottia pseudobrowniana G 66 Ostracoda Podocopida Cyprididae Stenocypris subterranea G + 67 Ostracoda Podocopida Cyprididae Cavernocypris subterranea G Loxoconchidae Pseudolimnocythere cf. 68 Ostracoda Podocopida hypogea G

106 69 Ostracoda Ostracoda Ostracoda Ostracoda gen. spp. G + + + 70 Ostracoda Podocopida Cyprididae Heterocypris oblonga G + 71 Ostracoda Podocopida Cyprididae Herpetocypris chevreuxi G + 72 Ostracoda Podocopida Cyprididae Humphcypris subterranea G + 73 Ostracoda Podocopida Cyprididae Humphcypris anomala G + 74 Ostracoda Podocopida Cyprididae Humphcypris brevisetosa G + 75 Ostracoda Podocopida Cyprididae Humphcypris chappuisi G + 76 Ostracoda Podocopida Cyprididae Humphcypris decipiens G + 77 Ostracoda Podocopida Cyprididae Humphcypris exigua G + 78 Ostracoda Podocopida Cyprididae Humphcypris greenwoddi G + 79 Ostracoda Podocopida Cyprididae Humphcypris leleupi G + 80 Ostracoda Podocopida Cyprididae Humphcypris sewelli G +

Appendix 1 (continued). References No.30 - No.59 (Table 1) Type Class Order Family Taxa 5 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 9

81 Ostracoda Podocopida Cyprididae Humphcypris thysvillensis G + 82 Ostracoda Podocopida Cyprididae Humphcypris tumens G + 83 Maxillopoda Cyclopoida Cyclopidae Acanthocyclops agamus G 84 Maxillopoda Cyclopoida Cyclopidae Acanthocyclops sp. G 85 Maxillopoda Cyclopoida Cyclopidae Diacyclops bicuspidatus G + 86 Maxillopoda Cyclopoida Cyclopidae Diacyclops copepodids G 87 Maxillopoda Cyclopoida Cyclopidae Diacyclops disjunctus G Diacyclops languidoides 88 Maxillopoda Cyclopoida Cyclopidae group G

107 89 Maxillopoda Cyclopoida Cyclopidae Eucyclops serrulatus G + 90 Maxillopoda Cyclopoida Cyclopidae Eucyclops sp. G + 91 Maxillopoda Cyclopoida Cyclopidae Megacyclops viridis G 92 Maxillopoda Cyclopoida Cyclopidae Macrocyclops albidus G + 93 Maxillopoda Cyclopoida Cyclopidae Macrocyclops sp. G + 94 Maxillopoda Cyclopoida Cyclopidae Itocyclops sp. G 95 Maxillopoda Cyclopoida Cyclopidae Speocyclops kieferi G 96 Maxillopoda Cyclopoida Cyclopidae speocyclops racovitzai G 97 Maxillopoda Cyclopoida Cyclopidae Paracyclops fimbriatus G + 98 Maxillopoda Cyclopoida Cyclopidae Paracyclops imminutus G 99 Maxillopoda Cyclopoida Cyclopidae Afrocyclops sp. G + Harpactico- Canthocampti- 100 Maxillopoda ida dae Attassaheyella crassa G Harpactico- Canthocampt- 101 Maxillopoda ida idae Atheyella nordenskioldii C + +

Appendix 1 (continued). References No.30 - No.59 (Table 1) Type Class Order Family Taxa 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 59 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 Bryocamptus (Arcticocamptus) 102 Maxillopoda Harpacticoida Canthocamptidae copepodites G

Bryocamptus 103 Maxillopoda Harpacticoida Canthocamptidae cuspidatus G 104 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus dacicus G 105 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus nivalis G +

106 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus rhaeticus G

108 Bryocamptus 107 Maxillopoda Harpacticoida Canthocamptidae vandouwei G

Bryocamptus (Bryocamptus) 108 Maxillopoda Harpacticoida Canthocamptidae pygmaeus G Bryocamptus (Echinocamptus) 109 Maxillopoda Harpacticoida Canthocamptidae ethinatus G Bryocamptus (Rheocamptus) 110 Maxillopoda Harpacticoida Canthocamptidae tatrensis G

111 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus typhlops G

112 Maxillopoda Harpacticoida Canthocamptidae Bryocamptus zschokkei G

Appendix 1 (continued). References No.30 - No.59 (Table 1) Type Class Order Family Taxa 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 59 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8

113 Maxillopoda Harpacticoida Canthocamptidae Cypria ophthalmica G + Elaphoidella 114 Maxillopoda Harpacticoida Canthocamptidae pseudophreatica G 115 Maxillopoda Harpacticoida Canthocamptidae Elaphoidella phreatica G 116 Maxillopoda Harpacticoida Canthocamptidae Epactophanes richardi G 117 Maxillopoda Harpacticoida Canthocamptidae Hypocamptus brehmi G Moraria (Moraria) 118 Maxillopoda Harpacticoida Canthocamptidae alpina G 119 Maxillopoda Harpacticoida Canthocamptidae Moraria poppei G 120 Maxillopoda Harpacticoida Canthocamptidae Moraria stankovitchi G 121 Maxillopoda Harpacticoida Canthocamptidae Moraria varica G 109 122 Maxillopoda Harpacticoida Canthocamptidae Stygepactophanes sp. G

Parastenocaridida 123 Maxillopoda Harpacticoida e Parastenocaris sp. G Harpacticoida gen. 124 Maxillopoda Harpacticoida Harpacticoida spp. G + 125 Malacostraca Amphipoda Seborgiidae Seborgia relicta G + Jesogammarus 126 Malacostraca Amphipoda Anisogammaridae fluvialis C Crangonyx 127 Malacostraca Amphipoda Crangonyctidae pseudogracilis G Crangonyx 128 Malacostraca Amphipoda Crangonyctidae subterraneus G 129 Malacostraca Amphipoda Crangonyctidae Crangonyx floridanus G 130 Malacostraca Amphipoda Crangonyctidae Synurella ambulans O

Appendix 1 (continued). References No.30 - No.59 (Table 1) Type Class Order Family Taxa 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 59 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 Crangonyctid- 133 Malacostraca Amphipoda ae Stygobromus russelli G + Crangonyctid- 134 Malacostraca Amphipoda ae Stygobromus gen. spp. G + 135 Malacostraca Amphipoda Gammaridae Echinogammarus gen. spp. G 136 Malacostraca Amphipoda Gammaridae Gammarus balcanicus G 137 Malacostraca Amphipoda Gammaridae Gammarus fossarum G + + 138 Malacostraca Amphipoda Gammaridae Gammarus lacustris G + 139 Malacostraca Amphipoda Gammaridae Gammarus pulex G + + 140 Malacostraca Amphipoda Gammaridae Gammarus minus G + + 141 Malacostraca Amphipoda Hadziidae Mexiweckelia hardeni G +

110 142 Malacostraca Amphipoda Hyalellidae Hyalella azteca G + Mesogammar- 143 Malacostraca Amphipoda idae Eoniphargus kojimai G 144 Malacostraca Amphipoda Niphargidae Niphargus aquilex G 145 Malacostraca Amphipoda Niphargidae Niphargus fontanus G 146 Malacostraca Amphipoda Niphargidae Niphargus foreli G 147 Malacostraca Amphipoda Niphargidae Niphargus laisi G 148 Malacostraca Amphipoda Niphargidae Niphargus sp. aff. puteanus G 149 Malacostraca Amphipoda Niphargidae Niphargus sp. G + 150 Malacostraca Amphipoda Niphargidae Niphargus gineti G 151 Malacostraca Amphipoda Niphargidae Niphargus gr. kochianus G 152 Malacostraca Amphipoda Niphargidae Niphargus gr. longidactylus G 153 Malacostraca Amphipoda Niphargidae Niphargus gr. stygius G 154 Malacostraca Amphipoda Niphargidae Niphargus tatrensis G Paraleptampho 155 Malacostraca Amphipoda pidae Paraleptamphopus sp. G +

Appendix 1 (continued). References No.30 - No.59 (Table 1) Class Order Family Taxa Type 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 Pseudocrangon- 156 Malacostraca Amphipoda yctidae Pseudocrangonyx sp. G 157 Malacostraca Amphipoda Pontogeneiidae Sternomoera sp. G 158 Malacostraca Decapoda Cambaridae Orconectes immunis G + 159 Malacostraca Decapoda Potamidae Geothelphusa dehaani C Proasellus deminutus 160 Malacostraca Isopoda Asellidae agg. O 161 Insecta Ephemeroptera Baetidae Baetis alpinus C 162 Insecta Ephemeroptera Baetidae Baetis vernus C + 163 Insecta Ephemeroptera Baetidae Baetis vagans C +

111 Maccaffertium 164 Insecta Ephemeroptera Heptageniidae meririvulanum C +

165 Insecta Ephemeroptera Heptageniidae Rhithrogena loyolea C 166 Insecta Odonata Calopterygidae Argia vivida C + 167 Insecta Odonata Calopterygidae Calopteryx atrata C 168 Insecta Odonata Calopterygidae Calopteryx sp. C + 169 Insecta Plecoptera Capniidae Capnopsis schilleri C 170 Insecta Plecoptera Chloroperlidae Alloperla mediana C + 171 Insecta Plecoptera Chloroperlidae Alloperla sp. C + Siphonoperla 172 Insecta Plecoptera Chloroperlidae burmeisteri C 173 Insecta Plecoptera Chloroperlidae Siphonoperla montana C 174 Insecta Plecoptera Leuctridae Leuctra braueri C + 175 Insecta Plecoptera Leuctridae Leuctra digitata C

Appendix 1 (continued). References No.30 - No.59 (Table 1) Class Order Family Taxa Type 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59

176 Insecta Plecoptera Leuctridae Leuctra tenella C + 177 Insecta Plecoptera Leuctridae Leuctra rosinae C 178 Insecta Plecoptera Leuctridae Perlomyia sp. C + 179 Insecta Plecoptera Leuctridae Leuctra juv. [group A] C 180 Insecta Plecoptera Leuctridae Leuctra juv. [group B] C 181 Insecta Plecoptera Nemouridae Amphinemura standfussi C 182 Insecta Plecoptera Nemouridae Amphinemura sulcicollis C 183 Insecta Plecoptera Nemouridae Protonemura meyeri C 184 Insecta Plecoptera Nemouridae Protonemura auberti C

112 185 Insecta Plecoptera Nemouridae Protonemura brevistyla C

186 Insecta Plecoptera Nemouridae Protonemura lateralis C 187 Insecta Plecoptera Nemouridae Protonemura nimborum C 188 Insecta Plecoptera Nemouridae Protonemura sp. C 189 Insecta Plecoptera Nemouridae Nemoura fulva C 190 Insecta Plecoptera Nemouridae Nemoura flexuosa C 191 Insecta Plecoptera Nemouridae Nemoura albidipennis C + 192 Insecta Plecoptera Nemouridae Nemoura cinerea C + 193 Insecta Plecoptera Nemouridae Nemoura completa C + 194 Insecta Plecoptera Nemouridae Nemoura obtusa C 195 Insecta Plecoptera Nemouridae Nemoura picteii C 196 Insecta Plecoptera Nemouridae Nemoura mortoni C

Appendix 1 (continued). References No.30 - No.59 (Table 1) Class Order Family Taxa Type 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59

197 Insecta Plecoptera Nemouridae Nemoura sinuata C 198 Insecta Plecoptera Nemouridae Nemoura vallicularia C + 199 Insecta Plecoptera Nemouridae Nemoura sp. C 200 Insecta Plecoptera Nemouridae Nemurella picteti C + 201 Insecta Plecoptera Notonemouridae Malenka flexura C + 202 Insecta Plecoptera Notonemouridae Spaniocerca zelandica C + + 203 Insecta Plecoptera Notonemouridae Spaniocerca sp. C + 204 Insecta Plecoptera Notonemouridae Cristaperla sp. C + 205 Insecta Plecoptera Notonemouridae Halticoperla viridans C +

113 206 Insecta Plecoptera Perlodidae Dictyogenus alpinum C

207 Insecta Plecoptera Perlodidae Dictyogenus fontium C 208 Insecta Plecoptera Perlodidae Diura nanseni C 209 Insecta Plecoptera Perlodidae Isogenus subvarians C + 210 Insecta Plecoptera Perlidae Acroneuria abnormis C + 211 Insecta Plecoptera Perlidae Eccoptura xanthenes C + 212 Insecta Plecoptera Perlidae Kamimuria quadrata C 213 Insecta Plecoptera Perlidae Isoperla albanica C 214 Insecta Plecoptera Perlidae Isoperla rivulorum C 215 Insecta Plecoptera Perlidae Perta sp. C 216 Insecta Plecoptera Peltoperlidae Peltoperla arcuata C + 217 Insecta Plecoptera Gripopterygidae Megaleptoperla grandis C + 218 Insecta Plecoptera Gripopterygidae Megaleptoperla sp. C +

Appendix 1 (continued). References No.30 - No.59 (Table 1) Class Order Family Taxa Type 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59

219 Insecta Plecoptera Gripopterygidae Taraperla howsei C 220 Insecta Plecoptera Gripopterygidae Zelandobius unicolor C + 221 Insecta Plecoptera Gripopterygidae Zelandobius pilosus C + 222 Insecta Plecoptera Gripopterygidae Zelandobius confusus C + 223 Insecta Plecoptera Gripopterygidae Zelandobius furcillatus C + + 224 Insecta Plecoptera Gripopterygidae Zelandobius illiesi C + 225 Insecta Plecoptera Gripopterygidae Zelandobius sp. C + 226 Insecta Plecoptera Gripopterygidae Zelandoperla decorata C + + 227 Insecta Plecoptera Gripopterygidae Zelandoperla sp. C + 114 228 Insecta Plecoptera Eustheniidae Stenoperla maclellani C + 229 Insecta Megaloptera Sialidae Sialis fuliginosa C 230 Insecta Megaloptera Corydalidae Archichauliodes diversus C + 231 Insecta Megaloptera Corydalidae Parachauliodes japonicus C 232 Insecta Megaloptera Nannochoristidae Nannochorista philpotti C + 233 Insecta Megaloptera Nannochoristidae Nannochorista sp. C + 234 Insecta Neuroptera Osmylidae Kempynus sp. C + 235 Insecta Neuroptera Osmylidae Osmylidae gen. sp. G 236 Insecta Trichoptera Apataniidae Apatania eatoniana O 237 Insecta Trichoptera Brachycentridae Brachycentridae gen. spp. O 238 Insecta Trichoptera Hydrobiosidae Psyllobetina attunga C + 239 Insecta Trichoptera Hydroptilidae Orthotrichia gen. spp. O 240 Insecta Trichoptera Philopotamidae Dolophilodes sp. C +

241 Insecta Trichoptera Philopotamidae Wormaldia copiosa C 242 Insecta Trichoptera Philopotamidae Wormaldia occipitalis C 243 Insecta Trichoptera Philopotamidae Wormaldia sp. C 244 Insecta Trichoptera Phryganeidae Ptilostomis sp. C + 245 Insecta Trichoptera Polycentropodidae Plectrocnemia conspersa C + 246 Insecta Trichoptera Polycentropodidae Plectrocnemia geniculata C + 247 Insecta Trichoptera Polycentropodidae Plectrocnemia sp. C Polycentropus 248 Insecta Trichoptera Polycentropodidae flavomaculatus C

115 249 Insecta Trichoptera Polycentropodidae Polycentropus sp. C + + + 250 Insecta Trichoptera Psychomyiidae Tinodes unicolor C 251 Insecta Trichoptera Rhyacophilidae Rhyacophila towadensis C 252 Insecta Trichoptera Rhyacophilidae Rhyacophila sp.X-2 C 253 Insecta Trichoptera Rhyacophilidae Rhyacophila sp.RD C Rhyacophila fasciata 254 Insecta Trichoptera Rhyacophilidae Hagen C 255 Insecta Trichoptera Rhyacophilidae Rhyacophila shikotsuensis C 256 Insecta Trichoptera Rhyacophilidae Rhyacophila lobifera C + 257 Insecta Trichoptera Rhyacophilidae Rhyacophila lezeyi C 258 Insecta Trichoptera Rhyacophilidae Rhyacophila impar C 259 Insecta Trichoptera Rhyacophilidae Rhyacophila transquila C 260 Insecta Trichoptera Rhyacophilidae Rhyacophila brevicephara C

261 Insecta Trichoptera Rhyacophilidae Rhyacophila brunnea C + 262 Insecta Trichoptera Rhyacophilidae Rhyacophila dorsalis C 263 Insecta Trichoptera Rhyacophilidae Rhyacophila nigra C 264 Insecta Trichoptera Rhyacophilidae Rhyacophila spp. C + + 265 Insecta Trichoptera Sericostomatidae Sericostoma personatum C + 266 Insecta Trichoptera Sericostomatidae Sericostoma sp. C + 267 Insecta Lepidoptera Pyralidae Nymphulinae gen.spp. C 268 Insecta Diptera Athericidae Atherix sp. C + 269 Insecta Diptera Chironomidae Diamesa bertrami C 116

270 Insecta Diptera Chironomidae Diamesa cinerella C 271 Insecta Diptera Chironomidae Diamesa latitarsis C 272 Insecta Diptera Chironomidae Diamesa gen. spp. C 273 Insecta Diptera Chironomidae Anzacladius sp. C 274 Insecta Diptera Chironomidae Boreochlus sp. C + 275 Insecta Diptera Chironomidae Metriocnemus fuscipes C 276 Insecta Diptera Chironomidae Parochlus sp. C 277 Insecta Diptera Chironomidae Prodiamesa bathyphila C + 278 Insecta Diptera Chironomidae Prodiamesa olivacea C 279 Insecta Diptera Chironomidae Prodiamesa sp. C + + + 280 Insecta Diptera Chironomidae Rheocricotopus atripes C

281 Insecta Diptera Chironomidae Rheocricotopus effusus C 282 Insecta Diptera Chironomidae Rheocricotopus fuscipes C 283 Insecta Diptera Chironomidae Stempellinella flavidula C 284 Insecta Diptera Chironomidae Stilocladius montanus C 285 Insecta Diptera Chironomidae Boreoheptagyia legeri C 286 Insecta Diptera Chironomidae Boreoheptagyia sp. C 287 Insecta Diptera Chironomidae Pagastia sp.1 C 288 Insecta Diptera Chironomidae Pagastia sp.2 C 289 Insecta Diptera Dixidae Dixa (Dixella) californica C +

117 290 Insecta Diptera Dixidae Dixa dilatata C

291 Insecta Diptera Dixidae Dixa inextricata C + 292 Insecta Diptera Dixidae Dixa marculata -agg. C + 293 Insecta Diptera Dixidae Dixa modesta C + 294 Insecta Diptera Dixidae Dixa naevis C + 295 Insecta Diptera Dixidae Dixa nipponica C 296 Insecta Diptera Dixidae Dixa neoaliciae C + 297 Insecta Diptera Dixidae Dixa submaculata C 298 Insecta Diptera Dixidae Dixa similis C + 299 Insecta Diptera Dixidae Dixa sp. C + + + + + 300 Insecta Diptera Dixidae Paradixa fuscinervis C + 301 Insecta Diptera Dixidae Paradixa harrisi C + 302 Insecta Diptera Dixidae Paradixa neozelandica C +

303 Insecta Diptera Dixidae Dixidae gen.spp. C + 304 Insecta Diptera Dolichopodidae Dolichopodidae n. det. C + 305 Insecta Diptera Psychodidae Pericoma albitarsis C + 306 Insecta Diptera Psychodidae Pericoma bipunctata C + 307 Insecta Diptera Psychodidae Pericoma spp. C 308 Insecta Diptera Psychodidae Psychoda parthenogenitica C + 309 Insecta Diptera Psychodidae Psychoda sp. C + + 310 Insecta Diptera Psychodidae Telmatoscopus albipunctatus C + 311 Insecta Diptera Psychodidae Telmatoscopus aldrichanus C +

118 312 Insecta Diptera Psychodidae Telmatoscopus basalis C +

313 Insecta Diptera Psychodidae Telmatoscopus bipunctata C + Telmatoscopus 314 Insecta Diptera Psychodidae quadripunctatus C + 315 Insecta Diptera Psychodidae Thornburghiella slossoni C + 316 Insecta Diptera Psychodidae Threticus bicolor C + 317 Insecta Diptera Psychodidae Tinearia alternicula C + 318 Insecta Diptera Chironomidae Pentaneurella sp. C + 319 Insecta Diptera Chironomidae Pentaneura sp. C + + + 320 Insecta Diptera Thaumaleidae Thaumalea testacea O 321 Insecta Diptera Thaumaleidae Thaumaleidae gen. spp. C 322 Insecta Coleoptera Dytiscidae Agabus biguttatus C 323 Insecta Coleoptera Dytiscidae Agabus bipunstulatus C

324 Insecta Coleoptera Dytiscidae Agabus didymus C 325 Insecta Coleoptera Dytiscidae Agabinus glabrellus C + 326 Insecta Coleoptera Dytiscidae Agabus guttatus C 327 Insecta Coleoptera Dytiscidae Agabus lapponicus C 328 Insecta Coleoptera Dytiscidae Agabus melanarius C 329 Insecta Coleoptera Dytiscidae Agabus spp. C + 330 Insecta Coleoptera Dytiscidae Antiporus stigosulus C + 331 Insecta Coleoptera Dytiscidae Coelambus sp. C 332 Insecta Coleoptera Dytiscidae Colymbetes sp. C

119 333 Insecta Coleoptera Dytiscidae Colymbetinae gen. spp. C

334 Insecta Coleoptera Dytiscidae Graptodytes sp. C 335 Insecta Coleoptera Dytiscidae Haideoporus texanus C + 336 Insecta Coleoptera Dytiscidae Hydroporus memnonius C 337 Insecta Coleoptera Dytiscidae Hydroporus nigrita C 338 Insecta Coleoptera Dytiscidae Hydroporus obsoletus C 339 Insecta Coleoptera Dytiscidae Hydroporus palustris C 340 Insecta Coleoptera Dytiscidae Hydroporus incognitus C 341 Insecta Coleoptera Dytiscidae Hydroporus tristis C 342 Insecta Coleoptera Dytiscidae Hydroporus sp. C + + 343 Insecta Coleoptera Dytiscidae Hydroglyphus sp. C + 344 Insecta Coleoptera Dytiscidae Huxelhydrus syntheticus C + 345 Insecta Coleoptera Dytiscidae Liodessus deflectus C +

346 Insecta Coleoptera Dytiscidae Platambus maculatus C 347 Insecta Coleoptera Dytiscidae Platambus sp. C 348 Insecta Coleoptera Dytiscidae Rhantus sp. C + Stictotarsus 349 Insecta Coleoptera Dytiscidae duodecimpustulatus C 350 Insecta Coleoptera Hydrophilidae Anacaena lutescens C 351 Insecta Coleoptera Hydrophilidae Anacaena globulus C 352 Insecta Coleoptera Hydrophilidae Aliplus sp. C 353 Insecta Coleoptera Hydrophilidae Ametor latus C + 120 354 Insecta Coleoptera Hydrophilidae Berosus affinis C 355 Insecta Coleoptera Hydrophilidae Berosus nigriceps C + 356 Insecta Coleoptera Hydrophilidae Berosus sp. C 357 Insecta Coleoptera Hydrophilidae Coelostoma orbiculare C 358 Insecta Coleoptera Hydrophilidae Helophorus brevipalpis C 359 Insecta Coleoptera Hydrophilidae Helophorus fuscipes C 360 Insecta Coleoptera Hydrophilidae Helophorus strigifrons C 361 Insecta Coleoptera Hydrophilidae Helophorus sp. C Hydrobius fuscipes 362 Insecta Coleoptera Hydrophilidae (Linnaeus) C Hydrobius fuscipes 363 Insecta Coleoptera Hydrophilidae (Fabricius) C 364 Insecta Coleoptera Hydrophilidae Hydrobius sp. C +

Appendix 2 Bethic invertebrate sampling in the Gamata basin (Research sites: Nebori Valley, Sodega Valley, Hiru Valley and Hiru Valley).

Order Family Japanese Name Taxon Nebori Nebori Sodega Sodega Hiru Hiru War i War i Total Lotic Lentic Lotic Lentic Lotic Lentic Lotic Lentic Ephemeroptera Ephemeridae フタスジモンカゲロウ Ephemera japonica 1 Ephemeroptera Leptophlebiidae トビイロカゲロウ属の 1 種 Paraleptophlebia sp. 2 2 Ephemeroptera Baetidae フタバコカゲロウ Baetiella japonica 1 10 1 Ephemeroptera Baetidae シロハラコカゲロウ Baetis thermicus 2 7 15 4 196 20 9 Ephemeroptera Baetidae ヨシノコカゲロウ Alainites yoshinensis 4 9 11 7 14 4 Ephemeroptera Baetidae トビイロコカゲロウ Nigrobaetis chocoratus 1 1 Ephemeroptera Baetidae トゲトガリコカゲロウ Tenuibaetis ursinus 1 Ephemeroptera Heptageniidae キイロヒラタカゲロウ Epeorus aesculus 6 Ephemeroptera Heptageniidae ユミモンヒラタカゲロウ Epeorus nipponicus 1 1 1 Ephemeroptera Heptageniidae ミドリタニガワカゲロウ Ecdyonurus viridis 1 1 Ephemeroptera Ameletidae ヒメフタオカゲロウ属 Ameletus sp. 1 8 18 ヨシノマダラカゲロウ 121 Ephemeroptera Ephemerellidae Drunella ishiyamana 2 15 2 Plecoptera Chloroperlidae ミドリカワゲラ科の数種 Chloroperlidae gen. spp. 3 1 2 13 3 1 37 19 19

Plecoptera Nemouridae オナシカワゲラ属の数種 Nemoura sp. 10 10 18 4 10 Plecoptera Nemouridae ユビオナシカワゲラ属の１種 Protonemura sp. 1 2 1 1 3 Plecoptera Nemouridae フサオナシカワゲラ属の１種 Amphinemura sp. 4 Plecoptera Perlodidae アミメカワゲラ科の１種 Perlodidae gen. spp. 2 3 5 Plecoptera Perlidae カミムラカワゲラ属の１種 Kamimuria sp. 1 1 Plecoptera Perlidae モンカワゲラ Calineuria stigmatica 1 5 1 Plecoptera Peltoperlidae ミヤマノギカワゲラ Yoraperla uenoi 1 4 1 Neuroptera Osmylidae ヒロバカゲロウ科の１種 Osmylidae gen spp. Trichoptera Philopotamidae ヒメタニガワトビケラ属の１種 Wormaldia sp. 2 Trichoptera Brachycentridae マルツツトビケラ属の一種 Micracema sp. 4 5 5 2 4 Trichoptera Lepidostomatidae オオカクツツトビケラ Lepidostoma crassicorne 2 3 2 Trichoptera Brachycentridae カクスイトビケラ Brachycentridae gen spp. 2 7 Trichoptera Arctopsychidae シロフツヤトビケラ属 Parapsyche sp. 3

Appendix 2 (Continuted).

Order Family Japanese Name Taxon Nebori Nebori Sodega Sodega Hiru Hiru War i

Lotic Lentic Lotic Lentic Lotic Lentic Lotic Trichoptera Hydropsychidae ミヤマシマトビケラ属 Diplectgrona sp. 7 4 Trichoptera Lepidostomatidae カクツツトビケラ属の数種 Lepidostoma sp. 5 14 Trichoptera Glossosomatidae ニチンカタヤマトビケラ Glossosoma nichinkata 12 Trichoptera Rhyacophilidae トワダナガレトビケラ Rhyacophila towadensis 1 2 4 Trichoptera Rhyacophilidae シコツナガレトビケラ Rhyacophila shikotsuensis Trichoptera Rhyacophilidae レゼイナガレトビケラ? Rhyacophila lezeyi Trichoptera Rhyacophilidae タシタナガレトビケラ Rhyacophila impar Trichoptera Rhyacophilidae トランスクィラナガレトビケラ Rhyacophila transquila Trichoptera Rhyacophilidae ヒロアタマナガレトビケラ Rhyacophila brevicephara Trichoptera Rhyacophilidae ナガレトビケラ属 X-2 Rhyacophila sp.X-2 1 7

122 Trichoptera Hydroptilidae オトヒメトビケラ Orthotrichia sp.

Coleoptrea Elmidae ヒメドロムシ亜科 Elmidae gen. spp. 1 Coleoptera Hydrophilidae ガムシ科の１種 Hydrophilidae gen. spp. 1 Coleoptera Scirtidae マルハナノミ属 Elodes sp. 1 2 Diptera Tipulidae モロフィルス属の 1 種 Molophilus sp. Diptera Tipulidae カスリヒメガガンボ属の１種 Limnophia sp. Diptera Tipulidae ウスバガガンボ属の１種 Antocha sp. 1 Diptera Athericidae クロモンナガレアブ Sragina caerulescence Diptera Empididae オドリバエ科の数種 Empididae gen. spp. 6 2 1 2 Diptera Sciomyzidae ヤチバエ科の 1 種 Sciomyzidae gen. spp. Diptera Dixidae ホソカ属の１種 Dixa sp. Diptera Psychomyidae チョウバエ科の１種 Pericoma sp. 12 Diptera Simulidae アシマダラブユ属の 1 種 Simulium sp. 2 4 2 Diptera Ceratopogoniidae ヌカカ科の１種 Bezzia sp. 1 3

Appendix 2 (Continuted).

Order Family Japanese Name Taxon Nebori Nebori Sodega Sodega Hiru Hiru War i War i Total

Lotic Lentic Lotic Lentic Lotic Lentic Lotic Lentic Trichoptera Hydropsychidae ミヤマシマトビケラ属 Diplectgrona sp. 7 4 7 Trichoptera Lepidostomatidae カクツツトビケラ属の数種 Lepidostoma sp. 5 14 5 Trichoptera Glossosomatidae ニチンカタヤマトビケラ Glossosoma nichinkata 12 6 12 Trichoptera Rhyacophilidae トワダナガレトビケラ Rhyacophila towadensis 1 2 4 3 Trichoptera Rhyacophilidae シコツナガレトビケラ Rhyacophila shikotsuensis Trichoptera Rhyacophilidae レゼイナガレトビケラ? Rhyacophila lezeyi Trichoptera Rhyacophilidae タシタナガレトビケラ Rhyacophila impar Trichoptera Rhyacophilidae トランスクィラナガレトビケラ Rhyacophila transquila Trichoptera Rhyacophilidae ヒロアタマナガレトビケラ Rhyacophila brevicephara Trichoptera Rhyacophilidae ナガレトビケラ属 X-2 Rhyacophila sp.X-2 1 7

123 Trichoptera Hydroptilidae カクヒメトビケラ属 Orthotrichia

Trichoptera Hydroptilidae オトヒメトビケラ Orthotrichia Coleoptrea Elmidae ヒメドロムシ亜科 Elmidae gen. spp. 1 Coleoptera Hydrophilidae ガムシ科の１種 Hydrophilidae gen. spp. 1 Coleoptera Scirtidae マルハナノミ属 Elodes sp. 1 2 Diptera Tipulidae モロフィルス属の 1 種 Molophilus sp. 1 Diptera Tipulidae カスリヒメガガンボ属の１種 Limnophia sp. 1 Diptera Tipulidae ウスバガガンボ属の１種 Antocha sp. 1 1 Diptera Athericidae クロモンナガレアブ Sragina caerulescence Diptera Empididae オドリバエ科の数種 Empididae gen. spp. 6 2 1 2 1 6 Diptera Sciomyzidae ヤチバエ科の 1 種 Sciomyzidae gen. spp. 1 Diptera Dixidae ホソカ属の１種 Dixa sp. 1 Diptera Psychomyidae チョウバエ科の１種 Pericoma sp. 12 1 Diptera Simulidae アシマダラブユ属の 1 種 Simulium sp. 2 4 2 2

Appendix 2 (Continuted).

Order Family Japanese Name Taxon Nebori Nebori Sodega Sodega Hiru Hiru War i War i Total

Lotic Lentic Lotic Lentic Lotic Lentic Lotic Lentic Diptera Ceratopogoniidae ヌカカ科の１種 Bezzia sp. 1 3 4 Diptera Tanypodinae モンユスリカ亜科の 1 種 Tanypodinae gen spp. 2 4 2 Diptera Diamesinae タニユスリカ属の 1 種 Boreoheptagyia sp. Diptera Diamesinae アルプスケユキユスリカ Pagastia sp. 2 Diptera Diamesinae オオユキユスリカ属の 1 種 Pagastia sp. 1 1 1 Diptera Orthocladiinae ツヤユスリカ属の１種 Cricotopus Nostococladius sp.（頭黒） Diptera Orthocladiinae サワユスリカ属 Potthastia sp. Diptera Orthocladiinae ハダカユスリカ属の１種 Caediocladius sp. Diptera Orthocladiinae ホソケブカエリユスリカ属 Neobrillia Kawai

124 Diptera Orthocladiinae ケバネエリユスリカ属の数種 Metriocnemus sp. Diptera Orthocladiinae エリユスリカ Orthocladius sp. 4

Diptera Orthocladiinae ヌカユスリカ属の一種 Thienenanniella sp. 2 2 Diptera Orthocladiinae ニセケバネエリユスリカ属の１種 Parametriocnemus sp. 7 6 1 13 Diptera Orthocladiinae テンマクエリユスリカ属 Eukiefferiella sp. Diptera Orthocladiinae ニセテンマクエリユスリカ属 Tvetenia sp. 1 1 3 3 5 6 3 13 Diptera Orthocladiinae シッチエリユスリカ属 Georthocladius sp. 3 3 Diptera Chironominae エダヒゲユスリカ属の１種 Cladotanytarusus sp. Diptera Chironominae ヒゲユスリカ属の 1 種 Tanytarsus sp. 1 14 38 4 18 53 Diptera Chironominae ハモンユスリカ属の 1 種 Polypedilum sp. 2 8 2 Diptera Chironominae ナガレユスリカ属の 1 種 Rheotanytarsus sp. 1 6 1 23 8