National Ecological Observatory Network (NEON) Domain 11...

National Ecological Observatory Network (NEON)

Domain 11 Core Tower, Soil Sampling Array, Terrestrial Sampling, Aquatic Array, and Aquatic Sampling at Lyndon B. Johnson National Grasslands

Proposal to USFS, Lyndon B. Johnson National Grasslands

National Ecological Observatory Network

NEON, Inc. 1685 38th Street Suite 100 Boulder, CO 80301 www.neoninc.org

NEON Proposal for Domain 11 Lyndon B. Johnson National Grasslands

This proposal has been prepared for the NEON, Inc. Domain 11 Core tower, terrestrial, and aquatic sites

located in the Lyndon B. Johnson National Grasslands and Pringle Creek. The proposed term for the

tower site, soil array, terrestrial sampling and aquatic array is approximately 30 years. The actual ground

disturbing activities will be limited to the tower foundation, instrument hut foundation, soil arrays, soil pit,

staging areas, access paths to the instrument hut and soil arrays, aquatic infrastructure, access paths

and stairs at Pringle Creek, power and communications runs, and small disturbances from terrestrial

sampling using beetle pitfall traps and collecting soil samples.

Purpose and Need for Facility

Justification The nation’s Grand Challenges in environmental science have been reviewed by the National Research

Council, the International Geosphere-Biosphere Programme, the Millennium Ecosystem Assessment,

Diversitas, and the US Climate Change Science Program. The Grand Challenges identify tasks within the

environmental science field requiring additional research to yield scientific and practical information for

the future. Three challenges have been identified as causes of change in continental – scale ecology

which could be addressed by a continental – scale observatory system: climate change, land use change

and invasive species.

The National Ecological Observatory Network (NEON/ NEON, Inc.) will establish a new level of ecological

science. It is the first ecological observatory network designed to test and develop ecological theory by

detecting and forecasting ecological change at continental scales over multiple decades. Such research

will enable a deep understanding of large-scale processes, feedbacks, and implications for both

ecosystems and human societies. Continuous NEON data streams, made possible by state-of-the-art

computing power and sensor and communications technologies, will be a national resource for ecological

research.

Objective

NEON is a non-profit, continental-scale observatory for discovering and understanding the impacts of

climate change, land-use change, and invasive species on ecology. NEON partitioned the United States

into 20 eco-climatic domains, each of which represents different regions of vegetation, landforms, climate,


and ecosystem performance. In each domain NEON will gather long-term data on ecological responses

of the biosphere to changes in land use and climate, and of feedback with the geosphere, hydrosphere

and atmosphere. The NEON Observatory is the first initiative in the biological sciences to be supported

through the National Science Foundation’s Major Research Equipment and Facilities Construction fund.

Each domain will host a fully instrumented core site in minimally managed “wild land” area slated to

operate for the 30-year lifetime of NEON, two relocatable sites related to land use, invasive species,

urban effects to operate for 10-12 years, and aquatic sites. The site proposed at Lyndon B. Johnson

National Grasslands is a proposed core site for Domain 11. The tower and aquatic sites are proposed to

be part of the NEON Observatory for the duration of 30 years.

Terrestrial Design Factors

Site Infrastructure Overview The Lyndon B. Johnson National Grasslands proposed NEON core site requires the installation of an

ecological observatory tower with guy line support, instrument hut, soil array, access paths, terrestrial

sampling plots and buried electrical and communications conduit. The project involves the collection of

data across the Lyndon B. Johnson National Grasslands property for a period of 30 years following the

installation of the scientific equipment. The entire tower, soil array, instrument hut, and path construction

survey boundary is approximately 2.25 acres.

The tower will be located at 33.401231, -97.570033 and the instrument hut at 33.401181, -97.569836.

The soil array will extend approximately 462 ft. from the tower along a perpendicular path. Ecological

samples and measurements will be collected through terrestrial sampling. Sampling plots will be

distributed throughout the Lyndon B. Johnson National Grasslands property and will be minimally

invasive. A 60% design review was conducted with the Lyndon B. Johnson National Grasslands staff on

August 12, 2014. On April 21, 2015 NEON personnel met with Lyndon B. Johnson National Grasslands

staff to walk the site and discuss any required design changes. This proposal reflects the changes

discussed during that April site walk. Please see appendix A for a general layout of the proposed tower

site.

Tower NEON proposes to construct a 72 ft. tall square lattice tower supported with four dead man guy anchors.

The four guy anchor foundations would be placed at an approximate depth of 10 ft. and will measure


approximately 6x8 ft., a small portion will be visible above grade where the guy wires connect to the

anchor. Each guy anchor will require a 20 ft. clear radius. The tower foundation footprint will be

approximately 8x8 ft. The tower will serve as a supporting structure for scientific sensors at various

heights above the ground. It will be equipped with an internal ship ladder system for access and

landings/working platforms for personnel observations and installations.

Instrument Hut

NEON proposes to install a prefabricated instrument hut to house electronic instrumentation, gas bottles,

tools, safety equipment, and other items for use during operations. The hut measures 8x20x9 ft. with a

10x20 ft. footprint. The foundation will be comprised of (6 to 8) 2x2 ft. concrete piers for the hut. The piers

will be 2 ft. deep, with a maximum underground footing width of 3.5 ft. The tower will be located

approximately 62.5 ft. from the tower to the East.

Example of a NEON instrument hut.

Example of a NEON tower with instruments.


Soil Array Soil arrays plots will be established off of the 462 ft. path extending south of the tower. There will be five

soil array plots measuring approximately 16.4x16.4 ft. located approximately 33 ft. from the path. Each

array will be accompanied by an electrical arbor. The arbors will measure 4.5 ft. tall with two sign posts

mounts. Each arbor will be accompanied by a device post with NEMA box to supply power. Each plot will

have several in ground and above ground sensors installed. Each belowground sensor will be installed in

a hole approximately 2.5 inches in diameter either vertical or at a 45 degree angle. The depth of the

sensor will depend on the particular measurement to be collected and the site, the maximum depth is 7 ft.

The soil array holes will be made using a portable Geoprobe machine. The sensors will collect data

including soil temperature, soil water content, soil CO2, minirhyzotron, soil heat flux plates, line quantum

and precipitation throughfall collector (a tipping precipitation gauge that is installed under the plant

canopy).

Precipitation Collection (DFIR) The Double Fence Intercomparison Reference (DFIR) is a primary standard precipitation collection

system that will be deployed at NEON Core tower sites. This assembly contains weighing-type

precipitation collector (triple redundancy), one metal-altar shield and two double-wooden octagonal

fences following U. S. Climate Reference Network (USCRN) specifications.

Example of a soil array arbor with on grade (non-trenched) conduit.


There will be two DFIRs at the Lyndon B. Johnson National Grasslands, one at the tower location and

one at the aquatic location. The tower DFIR will be located at 33.399417, -97.566975 and will be

surrounded by three fences. The USCRN required fences are wooden, measure 6 ft. high and are placed

in a 26 ft. diameter octagon. These fences act as wind shields and improve the ability to measure both

liquid and solid precipitation without influence from horizontal winds. In addition to the USCRN required

fences, NEON will also install a cattle panel fence to protect the tower DFIR and USCRN fences from

cattle. The precipitation collector will measure approximately 5 ft. tall and 4 ft. in diameter. The center

post and four supporting posts will be anchored with concrete. Up to 22 trees may be removed in the

vicinity of the DFIR to provide the necessary 52 m (170.6 ft.) buffer for the precipitation gauge.

Auxiliary Portals and Device Posts The auxiliary portal is a designated point near the edge of the tower observation area where the utility

company’s power and data services enter the site location. Device posts and portals provide

infrastructure to distribute power and communications to sensors on site. Device posts are approximately

4.3 ft. tall and 1 ft. wide. Portals are approximately 4.3 ft. tall and 3 ft. wide.

The tower auxiliary portal and a transformer will be located at approximately 33.401333, -97.564989

however, micrositing may occur with coordination of local utility company. The auxiliary portal will be

approximately 1633 ft. from the instrument hut.

NEON will install device posts with NEMA box enclosures to supply power. These posts are proposed

along the soil array arbors and independent sensors, such as the DFIR, to enable line of sight

communication between sensors. The NEMA box enclosures can be housed on the ground to minimize

visual impacts if required by the site host.

Example of a DFIR system; left to right: internal precipitation gauge and outer protective fencing.


Power and Fiber Distribution NEON will use power supplied by Wise Electric Cooperative. Underground utilities and communications

conduit will be trenched and completed with a standard walk-behind trencher to minimize impacts.

Extended overhead lines will be kept clear of trees by hand clearing saplings, as necessary, for the

duration of the NEON project.

Proposed power and communication conduit will run from the existing utility pole located at 33.393717, -

97.568017. The power and conduit lines will be trenched approximately 4,037 linear feet from the existing

utility pole, along the fence line and fire break to the proposed auxiliary portal (located approximately at

33.401333, -97.564989). Then the conduit will be trenched approximately 1,633 linear feet to the

instrument hut (located approximately at 33.401181, -97.569836), approximately 462 linear feet from the

hut to the end of the soil array, and also approximately 650 linear feet down a short stint of the horse trail

to the DFIR (located approximately at 33.399417, -97.566975).

Boardwalks, Paths and Improved Surfaces NEON proposes to lay gravel as needed for safe and reliable passage over the proposed power and

communication conduit/ access route running from the existing utility pole located at 33.393717, -

97.568017, along the fence line and fire break to the proposed staging and parking area located at

33.401306, -97.564889. The access route will be approximately 4,037 linear feet.

NEON proposes a 40x40 ft. staging and parking area. This area will be located at the entrance to the site,

adjacent to the auxiliary portal and fire break at 33.401306, -97.564889. The area will be improved with

woven geotextile and class 6 base course. It will become the long term parking area for operations staff to

reduce disturbance to the site.

NEON proposes to create an approximately 830 ft. long unimproved path from the staging area to the

edge of the construction boundary. In addition, technicians will walk down the existing horse trail to

access the DFIR.

NEON proposes to construct approximately 803 ft. long improved gravel path from the edge of the

construction boundary to the instrument hut. The path will be 4 ft. wide with construction constrains

extending 2 ft. parallel from the edge of the path. The path will be improved with class 6 base course.


NEON proposes to construct approximately 462 ft. long improved gravel path from the tower extending

the length of the soil array. The path will be 2.5 ft. wide with construction constrains extending 2 ft. from

the edge of the path. The path will be improved with class 6 base course.

NEON proposes to construct an aluminum boardwalk around the instrument hut and to the tower base.

The boardwalk will be approximately 128 ft. long by 3 ft. wide.

NEON proposes to mechanically remove the briar and nuisance vegetation within the tower construction

limits to ease construction and access to the site during construction. NEON will make every attempt to

avoid substantial trees during the removal process.

Fencing and Security

The soil horizon pit will be covered with plywood and surrounded with construction fence during the few

days the pit is open.

The tower DFIR will be enclosed with three fences. Two are U. S. Climate Reference Network (USCRN)

required wooden fences measuring 6 feet high and are placed in a 26 foot diameter octagon. The third

fence will be a cattle panel fence to protect the DFIR and wooden fences from cattle.

Two 8x4 ft. tube gates will replace the existing fence gate at the entrance of the public road to the site.

The existing horse gate will not be replaced.

Soil Horizon Pit A soil horizon pit will be dug 5x6 ft. wide, up to 7 ft. deep at 33.401411, -97.567275. The pit is used for

identifying the soil horizon characteristics and will have samples of dirt removed. The soil pit will be open

for approximately one week and will coincide with construction activities. While the pit is open it will be

covered with plywood and surrounded with construction fence as a safety precaution. Upon completion,

the pit will be re-filled with previously excavated dirt.


Terrestrial Sampling Terrestrial plot sampling is proposed to occur throughout the Lyndon B. Johnson National Grasslands

property boundaries. The sampling activity consists of collection methods that are considered to be low

disturbance including vegetation clipping, soil sampling, bird observations, small mammal trap-and-

release, gravid mosquito trapping, beetle pitfall trapping and tick drag sampling. Sampling will be

conducted within several different types of plots. Plots will be marked with PVC stakes and/or aluminum

disk markers flush with the ground. The USFS will be provided with list and map of possible plot locations

containing contingencies. NEON will work with the USFS to avoid any exclusion areas or known research.

Please see appendix B for a map of contingency terrestrial plot locations.

Approximately 30 tower plots: Tower plots measure approximately 20x20m and are located within

the tower’s airshed. Plant and soil samples will be collected from tower plots.

Approximately 30 distributed plots: Distributed plots include base plots, mosquito points, tick

transect loops, small mammal grids and bird grids.

o Base plots measure 40x40 m and host sampling for plants, beetles, and soils.

o Approximately 6 tick transect loops measuring 40x40m.

o Approximately 6 to 8 small mammal trapping grids measuring 90x90m.

o Approximately 10 to 12 bird observation walking grids measuring 500x500m, and

o Approximately 10 mosquito collection points.

Ground Beetles and Vertebrate Bycatch Shifts in ground beetle distribution and populations can indicate significant changes in the local ecological

community. Beetles will be collected using pitfall traps embedded in the ground. The cups are 7 cm in

depth and 11 cm in diameter. The traps will use a diluted solution of non-toxic propylene glycol to

Example of the soil horizon pit; left to right: initial assessment and sample collection.


preserve the samples for DNA analysis. The traps will be checked bi-weekly throughout the growing

season (typically April through September, but varies based on site) and will be removed when the

sampling season ends. Each pitfall trap is covered with a low clearance 20 cm x 20 cm hard plate cover

(approximately 1.5 cm above the lip of the cup) to mitigate non-target species collection. The cover

denies access to all but the smallest vertebrates; however, it is possible for very small amphibians,

reptiles, and mammals to still enter the trap. As an added precaution, a threshold of 15 individuals per

species, per plot, per season has been established. When 15 cumulative individuals of any given

vertebrate species are captured at a single plot in one sampling season, a problem ticket will be triggered

for consideration by NEON staff scientists. A copy of NEON’s IACUC protocols is available upon request.

Mosquitoes Mosquitoes are sensitive to climate variation and they are important as disease vectors (e.g. West Nile

virus and Dengue). Mosquitoes will be collected using CO2 baited light traps. These traps operate by

slowly releasing CO2, mimicking the exhaled breath of an animal. The CO2 and light attract mosquitoes

which are drawn into the trap by a fan. Mosquito traps will be located in each of the major vegetation

types.

Ticks Ticks will be collected to improve understanding of how the prevalence of infectious agents (e.g. Lyme

disease) changes over time within ecosystems. Nymphal and adult-stage ticks will likely be collected

March through November using a drag sampling method, pulling a small flannel 1m x 1m swath of cloth

across the ground. Sampling will occur at six different locations at each site.

Birds NEON will contract experienced field technicians to collect bird abundance and diversity data using

passive techniques of point counts involving only visual and auditory observations at pre-determined

locations. There will not be any collection or capture of bird species. Point counts involve a single

observer standing at a survey location for about 10 minutes with only binoculars, a laser rangefinder, and

a datasheet. The number of survey locations varies with the size of a site, ranging from 20 points up to

135 points. Surveys will be conducted within 20 minutes of the official sunrise time once or twice every

breeding season at each site. Timing of breeding varies across sites, but typically occurs from April - June

in much of the US. Technicians will conduct point counts over a minimum of 2 days up to a maximum of

21 days per year.


Small Mammals NEON will collect data about small mammals including diversity, reproductive condition, demography, and

seroprevalence in order to better understand how environmental and land use changes are affecting

populations over time, including the occurrence of the pathogens that infect them (e.g., hantaviruses,

Lyme disease). A mark-recapture study of small mammals will be conducted with blood collected from

target species for offsite pathogen screening and ultimate archiving at an accredited institution. Sherman

live traps (folding or non-folding, approximately 3 x 3.75 x 9 or 12 inches long) or Tomahawk live traps (5

x 5 x 16 inches long) will be used for trapping. Up to eight trapping grids will be deployed per site, with

three grids designated for blood collection. Each grid will consist of 100 live traps (10 m spacing – 10

rows – 10 columns). Traps will be set for three consecutive nights per bout every month at Lyndon B.

Johnson National Grasslands. Traps will be set as the sun sets and checked each consecutive morning

as soon as it is light enough to work.

All captured individuals will be marked with either one numbered ear tag (0.25 inches long, National Band

and Tag, Model 1005-1) or a subcutaneous, RFID PIT tag (pre-sterilized 8mm or 12mm - Biomark, Inc.).

Mode of tagging will be based on the length of the external pinnae, as some species, such as voles (e.g.,

Microtus spp., Myodes spp.) and pocket mice (e.g., Chaetodipus spp., Perognathus spp.), do not have

sufficient pinnae for securing ear tags. Ear tags are small, metal, and uniquely numbered (4 numbers

each). Ear tags do not cause unnecessary pain when applied quickly and with sharp and correct

application (using provided tool and placed in cartilage at base of external pinnae). PIT tags are injected

under the skin on the back of the animal. This placement does not impede movement, and tags do not

migrate from this location. In rare cases, if PIT tagging is deemed undesirable, considering the condition

of an animal (e.g., highly stressed), an animal may be released without tagging. Shrews will not be

tagged, but instead marked on the belly with a colored permanent marker, if not previously marked, and

release immediately. Individuals are marked to allow us to assess and study population dynamics,

reproductive condition, timing of colonization of new individuals, longevity and movement, and a variety of

other aspects of life history that require permanent individual marking. Rabbits, moles, gophers and

squirrels will be released with no handling.

Tissue samples to be collected will include one ear tissue sample from the untagged ear for genetic

analyses, using a sterile 2 mm tissue biopsy punch, and clipped whiskers and/or hair for isotopic analyses

to assess changes in water stress. Fecal samples will also be collected from captured individuals. Rabbits,

moles, gophers and squirrels will be released with no handling. Samples will not be collected from shrews.


Blood samples will be collected from target species. Samples will not be collected from Sciurids, Soricids,

Talpids, Geomyids, any individual weighing less than 10 grams, any individual with a pronounced or

physically debilitating injury, any individual that has already been captured and bled during a current

sampling bout, and/or any species excluded from bleeding based on permitting regulations. For additional

information regarding bleeding protocols please refer to the attached proposal.

All captured individuals will be processed according to the listed methodologies and released at the point

of capture. Large squirrels, gophers, weasels and rabbits will be released with no processing. Shrews will

be measured and identified and then released when possible. A copy of NEON’s small mammal IACUC

protocol is available upon requested.

Plants NEON will collect plant data to better understand how plant species vary seasonally and throughout the

years. Rare, threatened and endangered species will not be collected.

Aboveground biomass/productivity sampling

Clip harvest protocol – minimally destructive (results in collections)

-Herbaceous biomass clipped within distributed and tower plots

-1 clip per bout per plot from a 0.1m x 2 m strip; 2 clips per bout per plot if site is managed for grazing.

-Data collected 1 time per year, every 3 years at distributed plots

-Data collected 1-2 times per year, every year at Tower plots; data collected every 4 weeks in Tower plots

if site is managed for grazing.

-1 additional clip bout held every 5 years to address herbaceous biomass biogeochemistry. 1 clip per

selected plot from a 0.1m x 2 m strip. Four tower plots and 16 distributed plots will be sampled.

Special Case Protocol: Bryophyte harvesting - related to Clip Harvest protocol; only occurs if a site has

>20% moss/bryophyte cover across plots.

-minimally destructive (results in collections)

-Data collected 1 time per year in tower plots only

-1 clip per bout per plot from 0.1m x 2m strip

Plant Coarse Downed Wood protocol – tallying and bulk density sampling from nonliving downed logs

and branches (bulk density sampling results in collections)

-Measured/collected within distributed and tower plots

-Nondestructive tally measurements once every three years


-Bulk density samples collected once during first 3 years; collected once again 5 years after first collection

Plant Litter fall protocol – collection from nonliving plants (results in collections)

-Measured/collected within tower plots

-Elevated traps: Deciduous forest traps will be deployed year round and will be collected once in the

spring and once every two weeks during leaf senescence.

-Elevated traps: Mixed forest traps will be deployed year round and will be collected at least once per

month and once every two weeks during leaf senescence.

-Elevated traps: Conifer forest traps will be deployed year round and will be collected year round

-Ground traps: traps will be deployed year round – collected once per year.

Plant Measurement Veg Structure protocol – minimally destructive (no collections – tagging)

-Measured within distributed and tower plots

-Data collection 1 time per year, every three years at distributed plots

-Data collection 1 time per year, every year at tower plot

Plant Leaf area index protocol – nondestructive

-Measured/ collected within distributed and tower plots

-Data collection 1 time per year, every three years at distributed plots

-Data collection every 2 weeks during the growing season, annually at tower plots

Plant phenology protocol – nondestructive


-Data collection frequency depends on plants and growing season.

Plant diversity protocol – minimally destructive (results in collections)


-Initial collection of 20-40 common species to be collected, pressed, mounted.

-Non-destructive measurements made one or two times annually.

Belowground biomass sampling

Plant belowground biomass protocol – minimally destructive (results in collections)

-Measured/ collected within tower plots

-Collections one time per year, every five years.


Soils NEON will collect soil samples to analyze chemical, physical, isotopic, microbial, and biological properties.

These samples will also provide data for GIS and digital elevation models to create detailed soil maps for

the study sites. Sampling bouts will take place up to three times per year at approximately 10 plots

distributed throughout the site. Sampling events generally correspond with periods of high biological

activity, such as during peak greenness and seasonal transitions. At each plot, three soil samples will be

taken to a depth ranging from 30 cm to 1 meter, and at a diameter ranging between 3.5 cm to 11 cm. Soil

samples will be sent to a laboratory for analysis. Holes will be backfilled according to the property owner’s

specifications.

NEON also proposes a one-time site characterization soil sampling effort subcontracted through the

Natural Resources Conservation Services (NRCS). This assessment is an essential component of

NEON’s mission in understanding the biogeochemical processes at NEON sites. This important work

presents a unique opportunity for Lyndon B. Johnson National Grasslands, NEON and the public to

obtain and share valuable data on the physical and chemical properties of soil at NEON’s long-term

monitoring plots at Lyndon B. Johnson National Grasslands.

The NRCS is a service within the U.S. Department of Agriculture that has been characterizing and

sampling soils for over 100 years. The overarching objective of the NRCS is to conserve soil resources,

and it uses the highest standards when sampling soil. The NRCS has 12 regional offices that employ soil

scientists who are experts in local soil conditions. Through its Web Soil Survey program, it has mapped

soils in over 95% of the nation’s counties, and fulfilling this contract with NEON will add greatly to the

nation’s scientific knowledge.

The goal of this one-time effort is to characterize the physical and chemical properties of the soil

associated with the NEON sampling plots by conducting field observations of soil layers and sampling soil

layers for laboratory analyses. Soil sampling would only occur within NEON distributed and tower plots

previously evaluated by cultural resource contractors. After making field measurements of soil depth and

morphology, about one kilogram of soil material per soil horizon would be collected for laboratory tests.

The combination of field and laboratory measurements will provide critical ecological data for

understanding biogeochemical processes both at your site and across the NEON observatory.

Two soil sampling methods may be used: pit-excavation, and soil coring. The preferred method for soil

survey sampling is by pit excavation, because it both yields higher quality data and generally has a lower

visible impact. Soils often exhibit variable properties over distances and depths of just a few centimeters,

and pit excavation allows direct physical observation of soil horizon variability with depth and permits

accurate sampling of discrete soil horizons, which is critical for understanding overall soil quality and


fertility. Importantly, certain soil measurements such as soil bulk density cannot be made without pit

excavation. The bulk density measurement is essential for converting weight-based values to field

volumes, and for determining the site’s nutrient availability and carbon storage capability.

The standard methodology for soil surveying by pit excavation involves hand excavation of a small pit

approximately one meter square by one meter deep in size. Occasionally, if needed, an additional one

meter deep core will be taken from the floor of the pit for soil categorization purposes and then replaced.

On average, we expect that 15 to 20 plots would be sampled, but the number could range between 10 to

34 plots. At one square meter, a pit’s affected surface area is relatively small - less than 0.01% of the total

surface area of a 40m x 40m plot. During excavation, each soil horizon is carefully collected on a tarp and

kept separate from other horizons. Once sampling is complete, the hole is refilled with the excavated soil,

reassembling the original soil profile layer by layer and striving to ensure that even surface vegetation

remains intact. Because pit excavation technique reconstructs the pre-survey soil conditions, it is

considered less impactful than sampling by a coring device.

The alternative soil sampling method employs a soil coring device, which for this effort involves a hand

auger or spade. Hand augers of about 10 centimeters (4 inches) in diameter will be used to examine soils

to a depth of about one meter. Cored samples would be taken at the same number of plots as for the pit

excavations across the proposed NEON site. In order to obtain a similar amount of material as from pit

excavations, up to 16 core samples may be collected at each plot. After coring is completed, NRCS would

push in the upper edges of the hole to create a sealed surface, leaving behind a slight depression but no

visible hole. Through physical and biological processes the void beneath naturally fills in after a few

months to about a year, depending on soil type. Based on the small core diameter and the sealed surface,

this method should not present a risk to cattle or wildlife. While this is the preferred and most scientifically

sound mitigation strategy, if this method is deemed unacceptable, the NRCS could use other mitigation

procedures such as backfilling holes with clean, non-native soil material. Introducing non-native soil may

change the chemical and physical properties of the soil.


Site 1: Example of a NRCS soil pit during the City of Spokane Soil Survey.


Site 2: Example of a NRCS soil pit during an urban soil survey.


Airborn Observation Platform

The Airborne Observation Platform (AOP) is designed to measure the effects of land use change, and

changes in vegetation state and ecosystem functioning including the presence and effects of invasive

species through the use of airborne remote sensing technologies. Airborne spectroscopy and waveform

LiDAR will quantify plant species type and function, and vegetation structure and heterogeneity at the

scale of individual shrubs and larger plants. Panchromatic photography (30 cm resolution) will retrieve

fine-scale information on land use, roads, impervious surfaces, and built structures. The AOP is aircraft-

mounted and operated instrumentation and will not result in any ground impacts to the National

Grasslands. Flyovers are proposed to occur once a year. Flight paths have not been established, but will

be coordinated with staff at Lyndon B. Johnson National Grasslands and with local air traffic control prior

to any flights.

Aquatic Design Factors

Site Infrastructure Overview The Pringle Creek proposed NEON core aquatic site requires the installation of aquatic monitoring

instruments, meteorological instruments, access paths, wooden stairs, aquatic sampling and buried or

overhead electrical power and communications lines (to be determined). The project involves the

collection of data for a period of 30 years following the installation of the scientific equipment. Tall grasses,

shrubs or other vegetation may need to be trimmed back initially to install infrastructure and then

maintained as part of ongoing maintenance. Over the 30 year project it is also possible that new trees

may start to grow which would need to be removed. Please see Appendix E for a map showing

geographic locations of aquatic infrastructure and the proposed utilities run.

Aquatic Infrastructure Aquatic sensor 1 (S1) and aquatic sensor 2 (S2) will be installed at each end of the aquatic reach. S1 and

S2 are comprised of an approximately 9 ft. long pole with mounted sensors. Each pole will be anchored

into the substrate of the stream. S1 is proposed to be located at 33.378403, -97.783672 and S2 is

proposed to be located at approximately 33.378469, -97.781503. The sensors measure water surface

elevation, specific conductivity, dissolved oxygen, turbidity, chlorophyll a, fluorescent dissolved organic

matter (fDOM), photosynthetically active radiation (PAR), water temperature, and nitrates. The pressure

transducer, staff gauge and digital camera for determining discharge will be co-located with the S2 sensor.


DFIR: The Double Fence Intercomparison Reference (DFIR) is a primary standard precipitation collection

system. This assembly contains weighing-type precipitation collector (triple redundancy), one metal-altar

shield and two double-wooden octagonal fences following U. S. Climate Reference Network (USCRN)

specifications. The USCRN fences are wooden, measuring approximately 6 ft. high with the most exterior

fence placed in a 26 ft. diameter octagon around the precipitation collector. These fences act as wind

shields and improve the ability to measure both liquid and solid precipitation without influence from

horizontal winds. The precipitation collector will measure approximately 5 ft. tall and 4 ft. in diameter. The

center post and four supporting posts will be anchored with concrete. The Pringle Creek DFIR will be

located at approximately 33.377267, -97.781572.

Wet Deposition NADP: The National Atmospheric Deposition Program (NADP) sensor will measure water

chemistry of precipitation. The sensor will extend approximately 5 ft. above ground level. It will be

anchored in the ground with concrete at the depth of approximately 3.4 ft. The sensor is proposed to be

located at approximately 33.377252, -97.781659.

S1/S2 sensors with different anchoring depending on substrate; left to right: bedrock, cobble, sand.


Micromet: The aquatic meteorological station will measure approximately 12.3 ft. tall and 7 ft. in diameter.

It is proposed to be located at approximately 33.377200, -97.781783. The station will measure air

temperature, PAR (above water), incoming radiation (long/short wave), barometric pressure, wind speed

and direction, and relative humidity.

Groundwater monitoring wells: Eight groundwater monitoring wells are proposed at the following

coordinates in Table 1. Micrositing may occur during installation. Each well will vary in depth, depending

on the substrate. The actual well structure will measure approximately 2 inches in diameter and 3 ft. in

height. Each will be anchored with a 6x6 in. concrete footprint. Wells will be installed with a rotary auger.

Each well will contain a sensor to monitor water surface elevation, temperature, and specific conductivity.

Proposed wet deposition sensor.

Proposed aquatic meteorological station.


Well ID Latitude Longitude

D11-PRIN-OW-01 33.377834 -97.784363

D11-PRIN-OW-02 33.378255 -97.783752

D11-PRIN-OW-03 33.377535 -97.783518

D11-PRIN-OW-04 33.378305 -97.782651

D11-PRIN-OW-05 33.378313 -97.781759

D11-PRIN-OW-06 33.377739 -97.782197

D11-PRIN-OW-07 33.378722 -97.783856

D11-PRIN-OW-08 33.378823 -97.781677

Proposed groundwater well; left to right: side-view of installed well and aboveground design of outer casing.

Table 1: Well identification numbers and corresponding GPS coordinates.


Auxiliary Portals and Device Posts The auxiliary portals are located at a designated point near the edge of the aquatic observation area

where the utility company’s power and data services enter the site location. Device posts and portals

provide infrastructure to distribute power and communications to sensors on site. Device posts are

approximately 4.3 ft. tall and 1 ft. wide. Portals are approximately 4.3 ft. tall and 3 ft. wide.

The aquatic auxiliary portals will be located at approximately 33.377797, -97.7833. However, micrositing

may occur with coordination of local utility company. Wise County electric will run overhead power to the

auxiliary portals. After routed through the auxiliary portals, conduit power and communication lines will

transition to underground trenched power to the sensor device posts.

The device posts are proposed along the stream bank and adjacent to independent sensors to enable

line of sight communication between sensor arrays. The NEMA box enclosures can be housed on the

ground to minimize visual impacts if required by the site host. Locations of sensor supporting device posts

are as follows: aquatic micromet device post will be located approximately at 33.377255, -97.781783;

aquatic NADP/DFIR device post will be located approximately at 33.377252, -97.781603; aquatic S1

device post will be located approximately at 33.378353, -97.783672; and the aquatic S2 device post will

be located approximately at 33.378436, -97.7815.

Paths and Improved Surfaces NEON proposes to use an existing USFS road for access and parking near the aquatic site. The

proposed parking area is approximately at 33.376129, -97.781210. In order to park near this area NEON

respectfully requests gate and road access from the USFS so field techs may drive within 426.5 ft. of the

met station and DFIR.

NEON proposes trimmed native vegetative footpaths through the riparian habitat of Pringle Creek. An

unimproved footpath will lead to S1. Aluminum stairs or a ladder are proposed to access S1, it will likely

be located at approximately 33.378289, -97.783154. A second unimproved footpath will lead to S2.

Aluminum stairs or a ladder are proposed to access S1, it will likely be located at approximately

33.378379, -97.781519. Footpaths will require vegetation maintenance throughout the years which may

include cutting tall grass, trimming branches, or cutting new saplings. Please see Appendix D for the

design drawings of NEON’s aquatic site.


NEON also proposes to create a trimmed, unimproved, native footpath for groundwater access. The path

will likely be 3 feet wide. This path will likely be the access route for the ground water well drill.

Fencing and Security

To protect the site from intrusion and to ensure the safety of cattle, the aquatic DFIR, micromet sensor,

and wet deposition NADP may be enclosed by three strand barbed wire fencing.

The aquatic DFIR will be enclosed with three fences. Two are U. S. Climate Reference Network (USCRN)

required wooden fences measuring approximately 6 ft. high with the most exterior fence placed in a 26 ft.

diameter octagon around the precipitation collector. The third fence will likely be a three strand barbed

wire fence to protect the DFIR and wooden fences from cattle.

Aquatic Sampling and Measurements

NEON plans to measure water chemistry, reaeration, discharge, sediment, and the riparian area in

addition to sampling fish, aquatic macroinvertebrates, microbes, and aquatic plants at Pringle Creek.

Water Chemistry Water Chemistry samples will be collected up to 26 times per year. Each 4 liter sample will be collected

as a grab sample. Samples will be analyzed for general chemistry, anion and cations, dissolved and total

carbon and nutrients, stable isotopes of water and particulate carbon and nitrogen, and dissolved gases

(Carbon Dioxide, Methane, and Nitrous Oxide).

Microbes Microbes will be sampled in wadeable streams with water chemistry samples and scrub samples. Microbe

water sampling occurs 12 times per year with monthly water chemistry samples. Water samples will be

collected from surface water with a 4L bottle. Microbial scrub samples will occur 3 times per year, roughly

spring, summer, and autumn. Depending on the stream bottom, these samples will be collected by

scrubbing a small portion of 3-5 cobbles or pieces of wood with gloves, collecting 3-5 small grabs of sand

or silt sediment, or 3-5 small grabs of plant material. Technicians will remove any invertebrates from the

substrate prior to scrubbing. Microbes scrubbed from the cobble or wood will be collected into a tray and

filtered into a capsule filter, sand/silt samples will be places in a Falcon tube, and plant grab samples will

be placed in a Whirl-pak®. Microbe samples will be flash-frozen and sent to a lab for analysis.


Reaeration Reaeration (i.e., gas exchange) is the movement of oxygen from the atmosphere into the water, and is

measured as the net rate (i.e. gain and loss of oxygen) at which gas exchanges across the air-water

interface. Reaeration represents the net flux of O2 and CO2 into and out of the atmosphere, and

therefore accurately quantifying reaeration rates will be important for climate change research and carbon

budget calculations.

Reaeration is measured approximately six to 12 times per year by a simultaneous and continuous

injection of an inert gas, sulfur hexafluoride (SF6), or propane gas in a rare instance. SF6 is inert, has

minimal biological uptake and is deemed safe for aquatic life. The inert gas tracer is bubbled into an area

of slow-moving water in the stream. Approximate gas injection rates for SF6 are 100 mL/min for streams

with flow ~ 50-200 L/s and 200-300 mL/min for large streams >1000 L/s. To account for dilution due to

surface or groundwater inputs, a conservative solute tracer such as chloride (Cl-) or bromide (Br-) is

added to the stream in addition to the inert gas. Rhodamine may be used as the conservative tracer in

rare cases. The conservative tracer is added to the stream at a known rate (based on discharge), aiming

to increase the stream conservative tracer concentration 5-10 mg/L above background (i.e., pre-addition)

levels. Tracer addition needs to only be large enough to detect at the most downstream sampling site

and will vary by location, discharge and background conductivity.

Once the conservative tracer reaches a plateau (i.e., is no longer increasing) at the most downstream

sampling site (measured via a handheld electrical conductivity meter), water and gas samples are

collected at three to four locations downstream of the tracer addition, and the injection is stopped.

Discharge Flow, or discharge, will be studied as it directly affects the physical, chemical, and thermal attributes of a

stream’s ecosystem. NEON will monitor stage (height of water in the stream) to calculate discharge

continuously in all river and stream sites within the Observatory. The discharge data product will be a

crucial input to a number of additional high-level NEON data products, such as stream metabolism and

nutrient fluxes.

The NEON approach to obtaining a continuous record of stream discharge is via two complementary

methods used to generate a stage-discharge rating curve (SDRC). The SDRC is an empirical relationship

(formula) between a stream stage and the associated discharge at that stage which allows automated

measurements of stage to be converted to discharge. NEON staff will collect several concurrent

measurements of stream-level and discharge over a range of stream-levels and flow rates using an in-


stream pressure transducer and collecting measurements with a hand-held velocity meter during wading

surveys. Wading surveys will be conducted biweekly (approximately 26 times per year).

Sediment Sediment samples will be collected up to 5 times per year. Each sediment sample will be collected with a

scoop or hand corer from several deposition zones within the stream. A total volume of 5 liters of

sediment will be collected and analyzed by an external laboratory for trace elements, major metals,

organics, organic contaminants (PAHs/PCBs), and sediment grain size.

Riparian mapping Stream riparian and morphology mapping will occur in King’s Creek once per year during base flow and

at peak greenness. Riparian mapping tasks include recording vegetation composition and canopy cover

estimates. Additionally, bank angle, bank texture, and water height will be recorded.

Fish NEON proposes to collect fish from Pringle Creek. The data collected from fish sampling will provide

biodiversity information indicating ecosystem health, as well as length and weight which can indicate fish

condition or the health of the fish population. Fish are sensitive to changes in their environment and their

tolerance levels to these changes are well known for most species. Consequently, fish community data

can provide information on the health of the ecosystem.

DC or pulsed DC backpack electrofishing will be used to sample fish in the stream reach. The stream

reach is approximately 1 km in length and will be sampled with the electrofisher via a three pass depletion

study over the reach. Passes will be separated by no less than 30 minute intermissions. If electrofishing

is not effective, minnow traps may be used. Electrofishing-related injuries should affect < 1% of fish

captured. If this number is exceeded at the site, the technicians will stop sampling and contact the NEON

Aquatic Ecologist. Fish will be sampled two times per year, once in the spring and again in autumn.

Fish processing will occur after each electrofishing pass ends. Fish will be identified, weighed, measured,

and then released. Respiration will be monitored for anesthetized fish, then returned to water after

handling. Handling time per individual will be 10 minutes or less. Non-target species will be identified to

the lowest practical taxonomic level, photographed if possible, recorded, and then released without

handling. NEON will voucher unknown or unidentifiable individuals in the field (1-5 individuals per taxon);

these individuals plus any mortally injured individuals will be euthanized. Technicians will ensure that they

keep their hands clean and free of chemicals (i.e., insect repellent, sunscreen) and wet during handling. A

copy of NEON’s fish IACUC is available upon request.


Macroinvertebrates Stream benthic invertebrate communities are strongly affected by disturbance, including shifts in nutrient

and pollutant concentrations, physical disturbances such as floods, scouring, freezing, drought, and

biological disturbance such as predation. Benthic invertebrates will be sampled from riffles, runs, snags

and pools. Riffles will be sampled with a surber net, runs will be sampled with a surber net or kicknet,

snags will be sampled with a snag net, and deep pools will be sampled with a corer and/or petite ponar.

Invertebrate sampling occurs three times per year: spring, summer, and autumn. Sampling bouts will

always occur at or near baseflow conditions. Sampling will not occur directly following a flood in the

stream. Collected specimens will be euthanized, preserved in plastic sample jars, and sent to a laboratory

for analysis.

Aquatic plants and algae Macroalgae, bryophytes, lichens, and aquatic plants will be sampled three times per year to track

changes in abundance and diversity. Aquatic plants, bryophytes, and lichens will primarily be identified

in-situ along point transects and collected in quadrats for biomass measurements. Additionally, the

carbon and nitrogen composition of plants and bryophytes will be measured. Biomass samples will be

processed (identified and weighed) by NEON domain staff at the Domain Lab, any specimens that cannot

be positively identified at the domain lab will be sent to external taxonomists. NEON will not collect

endangered species if present.

For the purposes of analysis, algae can be broken into three distinct groups: macroalgae, benthic

microalgae (periphyton), and sestonic algae (algal cells sampled in the water column). Macroalgae will

be sampled as part of aquatic plant, bryophyte, lichen, and macroalgae sampling. Periphyton will be

sampled from riffles, runs, and pools. The majority of the periphyton community may be colonizing the

leaves of aquatic macrophytes (epiphytes) or woody debris at some sites, thus these substrata are

sampled rather than sampling scarcely populated sandy substrata. Aquatic plant and algae sampling

occurs three times per year: spring, summer, and autumn. Sampling bouts will always occur at or near

baseflow conditions. Sampling will not occur directly following a flood in the stream.


Site Access and Activities

Construction The NEON facilities construction team, approximately ten people comprising both of NEON, Inc.

personnel and its contractors, will require access to the site for the duration of the construction period.

Construction typically lasts four to six months. NEON will employ a Construction Site Supervisor to

oversee all contractor actions, implement health and safety protocols, and track daily activity logs at the

site. Every NEON site will be protected to the maximum extent possible during construction. Strict

guidelines and considerations will apply to materials used in construction to minimize their impact on the

environment. NEON will employ appropriate BMPs in all construction design and activities.

Typical construction equipment may include: mini-excavators, skid-steer and pickup trucks. Typical

construction staff include: NEON construction and engineering staff, NEON safety and permitting staff,

NEON scientists, electrical contractors, foundation contractors, the tower and instrument hut assembly

crew, and inspectors.

Operations Once in operation, each NEON site will be managed by a Manager of Field Operations/ Domain Manager,

who will have a supporting staff of approximately five full-time-equivalent (FTE) personnel and multiple

seasonal field technicians. The Domain Manager will be the point-of-contact (POC) for each site’s land-

owner. The tower and aquatic sites will be visited weekly. NEON may request key access to gated areas

of the National Grasslands if the tower or plots are accessible through a locked gate. The Domain Manger

will coordinate sampling schedules with the Forest Service.

Rehabilitation and Reclamation After the 30 year span of the Domain 11 Core Site, NEON will return the site to USFS requirements. The

existing tower and infrastructure will be disassembled and removed. concrete foundations will be left in

place.


Appendix A

100% design drawings of NEON tower.


Appendix B

Proposed NEON tower infrastructure and power/ access.


Appendix C

Proposed tower and contingency plot locations.


Appendix D

100% design drawings of NEON aquatic site.


Appendix E

Proposed Pringle Creek infrastructure and power run locations.

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