03 Sol Att A Statement of Work.pdf

STATEMENT OF WORK

National Centers for Coastal Ocean Science

Monitoring and Assessment Branch

Analytical Chemistry Programs

FY2025

National Oceanic and Atmospheric Administration (NOAA)

National Ocean Service (NOS)

National Centers for Coastal Ocean Science (NCCOS)

Stressor Detection and Impacts Division (SDI)

Monitoring and Assessment Branch (MAB)

Mention of products or trade names does not constitute endorsement or recommendation for their use by the United

States government.

TABLE OF CONTENTS

1. PROGRAM MANAGEMENT

1.1. Project Description and Background

1.2. Task Description

1.3. Accreditation Requirements

2. FIELD SAMPLING

2.1. Mussel Watch

2.2. Bioeffects

2.3. Sample Collecting and Processing

2.3.1. Water Quality Indicators

2.3.2. Composited Surficial Sediment

2.3.3. Composited Bivalve Samples

2.3.4. Bioeffects and special study sampling

2.4. Sample Handling and Shipping

2.5. Documentation Requirements

2.5.1. Chain of Custody

3. LABORATORY ANALYSES

3.1. Sample Disposition

3.2. Analytical Methods Requirements

3.2.1. Chemical Contaminants

3.2.2. Contaminants of Emerging Concern (CECs)

3.2.3. Biotic Health Indicators

3.2.4. Water Quality Parameters

3.2.5. Sediment Particle Size

3.2.6. Total Organic Carbon

3.2.7. Histopathology

3.2.8. Bioassays

3.3. Quality Control Requirements

3.3.1. Measurement Quality Objectives

3.3.2. Instrument/Equipment Testing, Inspection, and Maintenance

3.3.3. Data Validation and Usability

3.4. Data Management and Reporting

3.4.1 Sediment, Tissue, and Water Contaminants

3.4.2. Sediment Characterizations

3.4.3. Water Quality Parameters

3.4.4. Bivalve Characterizations

3.4.5. Bioassays

4. ASSESSMENT/OVERSIGHT

5. REFERENCES

6. APPENDICES

Appendix 1. MWP Target Collection Dates.*

Appendix 2. NS&T Data Format Key and Information

Appendix 3. Chemical Analyte Qualifier Codes

Appendix 4. Data Formatting Templates

Appendix 5. Evaluation of Selected Biomarkers

Appendix 6. Sediment Toxicity Assessment

Appendix 7. Example Chain-of-Custody Form

Appendix 8. Example Field Data Collection Form

1. PROGRAM MANAGEMENT

1.1. Project Description and Background

The National Oceanic and Atmospheric Administration’s (NOAA) National Centers for Coastal Ocean Science

(NCCOS) seeks to continue its long-term nationwide monitoring and assessment efforts to survey the condition of the nation’s coastal ocean resources. NCCOS’ Monitoring and Assessments Branch (MAB) National Status and

Trends (NS&T) Program is designed to describe the current status of, and detect changes in, the environmental quality of our nation’s estuarine and coastal environments. Using a combination of targeted and probabilistic sampling designs, a common set of indicators along with an expanding list of analytes shall be utilized to survey and assess the condition of coastal resources, and subsequently incorporated into NS&T’s existing database. The data collected by the Program are provided to regional, federal, state and local resource managers and the public via publications and online access to the data and information products. The Mussel Watch and Bioeffects Assessment

Programs, along with various Special Studies constitute NS&T.

The purpose of the Mussel Watch Program (MWP) is:

1) To monitor the status and trends of toxic chemical accumulations in the nation’s coastal and Great Lakes waters. The program is based on the collection of sediment and bivalve shellfish samples from a network of monitoring sites, and the analysis these sample for a suite of inorganic elements (i.e., metals), organic compounds (i.e., pesticides, PCBs) and emerging contaminants (i.e., PFAS). Currently the program’s sampling follows a rotating regional schedule, with a five-year cycle for each of the five regions, Gulf of

Mexico (GOM), Great Lakes (GL), North East (NE), South East (SE), and West Coast (WC).

2) To provide coastal resource managers and public with data and data products for the evaluation of assessing local and regional coastal conditions, enabling comparisons with national data.

The purpose of the Bioeffects Assessment Program is:

1) To provide spatially intensive toxicity testing, benthic biological community condition assessment, and chemical contaminant analysis in selected coastal ocean resources and sediment in order to describe the nature and scale of biological effects;

2) To provide resource managers with data to evaluate the success of coastal resource management strategies or effects of coastal impacts (land-based sources of pollution, energy exploration, etc.); and

3) To complement the programs of other federal and state agencies. Bioeffects Assessments are place-based typically conducted as discrete site/region characterizations, and field work is generally performed by

NCCOS in cooperation with local partners.

Special projects are sometimes conducted to address targeted resource management needs. These may be conducted in collaboration with other portions of NCCOS, with other federal agencies, or state/local partners. Special Studies may use the standard NS&T analytical suite, a reduced scope of analytes, or contaminants not measured as part of the standard suite. Similarly, Special Studies may sometimes use other tissue matrices beyond the standard biota

(i.e., bivalves, fish, and coral) used by NS&T.

1.2. Task Description

This Statement of Work (SOW) details the tasks required to carry-out national field sampling efforts, to precisely and accurately measure environmental stressors (e.g., trace metals and organic contaminants, nutrients, nanoparticles, plastic and microplastic particles, personal care products and pharmaceuticals, biomarkers, microbial assays (e.g., fecal coliform), and omics in various environmental matrices (e.g., sediment, water, and biological tissue)), and to provide deliverables in a timely manner to submit proposals.

The primary task for this contract is to provide analytical chemistry services for NCCOS’s Mussel Watch, Bioeffects

Assessment, and Special Studies Programs. Secondarily, NCCOS may contract all or a portion of field sampling activities, particularly for Mussel Watch. The period of performance of the contract shall be five years divided into five options. The Contract Scientific Lead shall interact periodically with the scientific and field staff of the successful laboratory to ensure scientific standards are met.

1.3. Accreditation Requirements

The contractor must maintain the following Laboratory Accreditations:

ISO/IEC 17025 Testing and Calibration: Specifies general requirements for the competence to carry out test and calibrations, including sampling. This accreditation also covers testing and calibration quality systems using standard methods and laboratory-developed or modified methods.

ISO 9000 Series of Standards: A set of international standards on quality management and quality assurance developed to help companies effectively document the quality system elements needed to maintain an efficient quality system. They are not specific to any one industry and can be applied to organizations of any size.

Furthermore, during the course of the contract, NOAA may elect to partner with another agency on a specific project, whose laboratory is also analyzing samples for chemical contaminants. At that time, the Mussel Watch contractor may be asked to participate in an interlaboratory comparison, in order to assess the comparability of the analyses between the two laboratories.

2. FIELD SAMPLING

The NS&T’s MWP is an environmental monitoring program that utilizes both bivalve shellfish and surficial sediments. Mussel Watch sites are not randomly selected but instead located where sufficient mussel or oyster beds are found. Sampling for the program occurs on a regular, rotating basis. Over the 30-year duration of MWP, field sampling has been conducted by contract labs, NCCOS, and local partners. NS&T’s Bioeffects Assessment Program is an environmental assessment and research program customized to the needs of specific estuaries, the Great Lakes, and US coastal waters in order to provide information needed to evaluate, enhance, or develop resource management strategies.

All field crews that participate in field sampling shall utilize the standard collection protocols described for Mussel

Watch and for Bioeffects Programs at https://coastalscience.noaa.gov/. NCCOS may choose to require training for new field crews or in the event that new protocols are required for either standard NS&T programs or Special

Studies. These training sessions would be organized and presented by the NS&T staff or their associates. Field training may be held either at individual sites or at a centralized location. During the training, crews would be instructed on sampling protocols and methods developed for NS&T, then they shall actively participate in hands-on exercises conducted in the field during which all components of the field sampling shall be evaluated by the instructors on a pass/fail basis for each component as they conduct a full NS&T field sampling scenario.

Permits and Permissions

All Mussel Watch sites in coastal waters lie within three nautical miles (nm) of shore. Hence in general, they fall within the authority of coastal states but in some instances, there is shared responsibility with the Federal government. The US Territorial Sea extends from three to 12 nm from shore, with the Federal government having sovereignty over resources in this area. Permission to collect living resources and in some instances permission to collect sediment is required from authorities with jurisdiction.

The contractor shall adhere to all federal and state laws when accessing sites, collecting samples, and transporting samples in the field. The contractor shall obtain all necessary permits including federal and state permits required for the collection of samples. Generally, these are scientific collection permits issued by the state. In some instances, federal permits may also be required when sampling on federal property or collecting organisms out of season.

The contractor shall be solely responsible for timely submission of permit applications, permit fees, and end of permit reporting. Any violation of federal or state laws or regulations including permit violations is the responsibility of the contractor. The contractor shall work through state and/or federal agencies or resource management commissions to ensure that sampling activities on leased shellfish beds are pre-approved.

2.1. Mussel Watch

Presently there are about 300 MWP sites located throughout the contiguous US, Alaska, Hawaii, and Puerto Rico

(Figure 1). Since its launch over 30 years ago, the program has maintained consistent quality assured sampling protocols. However, sampling frequency has changed over the years, and currently, the program has adopted a rotating regional monitoring approach that provides actionable information to regional stakeholders, while preserving its national foundation. In the new approach, bivalves and sediment are collected at historical sites, but also at new-targeted sites in order to assess specific regional needs. Existing Mussel Watch site descriptions can be https://coastalscience.noaa.gov/ found in NOAA Technical Memorandum NOS ORCA 112 which is available upon request from NCCOS. Limited site location and descriptions can be accessed via the NS&T web portal at https://products.coastalscience.noaa.gov/collections/ltmonitoring/nsandt/default.aspx.

Figure 1. Map of historical Mussel Watch sites with the associated number of years sampling has occurred for each site (one sample per year).

Field crews shall access Mussel Watch sites by the best practical means, which may include automobile (walk-up sites), boat, or aircraft. Site visits and sample collection shall occur within a pre-determined sampling window

(several weeks); samples are currently collected in fall/winter (September-March) so that the bivalve lipids shall be at post-spawning levels. An example of target sampling dates can be found in Lauenstein et al., 1997. The field teams shall be provided with a list of sampling sites and target latitude and longitude by NCCOS. The crew shall locate the sites using Wide Area Augmentation System (WAAS) enabled Global Positioning System (GPS).

Agreement between the given coordinates and the actual in-the-field siting of a subtidal sampling station should be within 0.02 nm, which is equivalent to a radius of approximately 40 meters (m). In cases where the vessel cannot navigate to within 0.02 nm of the intended site (e.g., the site is actually landlocked or the depth too shallow), the crew shall record the station as “intended-unsampleable” and thoroughly document the reason(s) on the Station

Information Data Sheet. The crew may then access the site by foot if there are bivalves there to sample, or relocate the vessel to the nearest position that permits sampling and conduct the monitoring, recording the coordinates and other characteristics of the relocated site. When this issue arises, the contractor shall contact the Contracting

Officer’s Representative (COR) and MWP Program Manager (PM) prior to leaving the site for further direction.

Intertidal sampling locations (walk-up sites) that are being revisited should be sited within the accuracy provided by the GPS unit, or approximately 5m.

Within each site, three independent samples (stations) shall be taken. These station samples shall be packaged separately and returned to the laboratory. A subset of samples from each station at a site shall then be composited in the laboratory. The intent of compositing three stations per site is to take inter-station variability into account with each analysis. Offshore subtidal sites are 400m radius circles with the center of that circle being the given latitude and longitude for that site. It is within this 400m radius that the three stations per site shall be collected. Shoreline intertidal sites are defined as a 100m linear distance along a tidal horizon to either side of the site center (local conditions may restrict this distance to less than 100m).

Sampling substrates are limited to rock (including rip-rap and jetties), concrete walls, sand, and mud. No samples collected as part of the regular MWP work shall be taken from structures whose composition could be a source of chemical contamination; for example, wood pier pilings and navigation aids shall be avoided.

Establishing new sites

The project manager may direct a field team to establish a new monitoring site based upon prior knowledge before the field crew enters the field. However, upon visiting a pre-existing site the field crew may find that no live bivalves are present and the site is non-viable. Also, in cases where conditions have changed and an existing site becomes devoid of viable oyster beds, at the direction of the MWP PM in charge of oversight and COR, the field https://products.coastalscience.noaa.gov/collections/ltmonitoring/nsandt/default.aspx crew may be instructed to evaluate relocating the site to nearby locations where bivalves are available. A detailed description on how to establish a new Mussel Watch site is provided in Apeti et al. (2012)

Over the course of this multi-decade program some sample sites have been lost due to a number of factors including but not limited to subsidence, destruction of substrate (i.e., pier or seawall), and die-off of mollusk beds. If a field crew finds a previous sampling location where it is no longer possible to collect samples, the crew shall photo document the conditions at the site along with a written description of the conditions which do not allow sampling to occur and provide both electronically to NCCOS. New sites may be established to replace lost ones in coordination with the MWP PM.

Mussel Watch sampling

The SOP for collecting bivalve samples for the MWP is described in Apeti et al. (2012). Field activities performed at each site should require approximately one to two hours per site. The number of sites sampled per day is subject to such factors as weather, seas, ease of access, and travel distance.

Sampling effort for each site varies depending on sampling techniques required. In addition, equipment and transportation necessary to get to the site can differ substantially as well (Table 1).

Table 1. Transportation and accouterments needed for sample collection*.

Transportation needed Percentage of sites Sampling gear Percentage of sites

Automobile (walk-up) 48 Hand collection 76

Boat 51 Diving (snorkeling) 5

Float plane - Day boat trip 1 Dredging 19

*Note: This information is a summary of all of the Mussel Watch sites. Currently all Mussel Watch samples are collected by NCCOS personnel and collaborating organizations. It is anticipated that these arrangements shall continue into the future.

At each sampling site, all crews shall uniformly collect a core set of data and samples following NCCOS methods and protocols (see section 2.3.1 below). Core field data/samples include: dissolved oxygen, pH, salinity, temperature, depth, transmittance, clarity, actual latitude and longitude, weather conditions, and observations on tides.

Sediment sampling

The SOP for collecting bivalve samples for the MWP is described in Apeti et al. (2012). Sediment sites shall be coincident with bivalve sites. Every effort is made to maintain the sediment sampling site as near to the bivalve sites as possible, preferably within the defined bounds of the bivalve site. Sediment sites are depositional and subtidal

(never exposed at lowest low tides; note: sediments are not to be collected at low tide in the intertidal zone using only a sediment scoop). Surficial sediment should contain at least 20% fine-grained material (≤ 64 microns) on a dry-weight basis. Samples that do not meet these grain-size criteria shall not be collected for contaminant analysis unless approved by the COR. Sediment sites should be exposed to the same water mass as the corresponding bivalve site. Sediment sites that are an exception to this have been/shall be identified with a separate four-letter site acronym. If acceptable sediment cannot be located within the established sediment site, the COR shall be contacted and relocation of the sediment site may be discussed and agreed to.

The COR shall specify through written or electronic correspondence which sites shall be sampled for bivalves and/or sediment and when. Delivery orders issued from this contract will detail the specific list of sites to be sampled from. Sites shall be collected within three weeks of the target dates identified in Appendix 1 unless mutual agreement between government and contractor.

Field teams shall notify the COR and CO if a site cannot be collected so that an alternate or new site can be established. Problems in sample collection that would require compromising the site criteria shall be brought to the immediate attention of the COR by sampling crews or principal investigator, for joint resolution. This includes changing the existing designated site species or finding suboptimal sediment characteristics. If it is determined by the field team that a minor relocation or abandonment of the sampling site is required, the COR shall be contacted immediately and approval shall be obtained prior to any sampling effort at a new location. The decision to relocate sites remains with the COR. If another location is chosen for sampling, a determination shall be made by the COR as to whether the new location shall be considered a minor “relocation” of the site or whether the new site is significantly different and should be considered a newly established site. New sites, established during each sampling year, shall be assigned new site names and unique site codes. Sites minimally relocated may be renamed and receive a new four-letter code if this change would better signify the relocated site; this is also at the discretion of the COR and CO.

2.2. Bioeffects

Bioeffects Assessments are customized to the needs of specific geographical areas of concern and use a combination of sediment chemistry, benthic infauna characterization and sediment toxicity bioassays which comprise the

Sediment Quality Triad approach (Chapman et al., 1987). Bioeffects Assessments are generally a one-time intensive sampling effort that may include both stratified random and targeted sampling.

Bioeffects Assessments and Special Studies can include both targeted and probabilistic sampling designs. All randomly selected sites shall include at least two alternate sites. Field crews shall strictly adhere to the guidelines for siting the station unless there are substantiated reasons that prevent sampling within that defined area. Most studies are designed based on a stratified random sampling approach that uses Geographic Information System (GIS) technology to probabilistically generate sampling locations. Typically, these studies have greater sampling intensity than most Environmental Monitoring and Assessment Program (EMAP)-like studies. Results from such a study can be found in Hartwell and Hameedi (2007).

Surface sediment and/or tissues shall be analyzed for the same contaminants as in the MWP. The approach for sampling is different as Bioeffects Assessments require additional samples including macrobenthic community characterization and sediment toxicity assays. A detailed description of the SOP for field sampling is provided in

Apeti et al. (2012).

2.3. Sample Collecting and Processing

NCCOS samples require working with a variety of physical and biological measurements as follows:

2.3.1. Water Quality Indicators

The first activities that should be conducted upon arriving onsite for NS&T are those that involve water sampling and water column measurements; these samples/data need to be collected before disturbing bottom sediments.

Core water parameters include dissolved oxygen, pH, salinity, temperature, depth, transmittance (Secchi depth), and clarity. Salinity measurements shall be taken at each site. Salinity may be determined with the use of an electronic instrument such as a casting of conductivity, temperature and depth sensors instrument (CTD), or by refractometer reading of the actual water sample taken by Van Dorn/Niskin etc. For any instruments used, NCCOS requires documentation that the instruments have received the manufacturer’s suggested calibration and quality assurance checks.

The water column shall be sampled at each site for the determination of dissolved nutrients (N and P species), chlorophyll concentration, and total suspended solids by using a Van Dorn sampler or Niskin bottle. Water samples shall be collected as follows:

1. Shallow sites (< 2m) – sample at 0.5m (near-surface) and 0.5m off the bottom;

2. Standard site (> 2m) – sample at 0.5m (near-surface), mid-depth, and 0.5m off the bottom;

3. Unless the depth is so shallow that the near-surface and near-bottom overlap; then sample mid-depth only.

An approximately 3-Liter (L) subsample shall be pulled into a clean wide-mouth Nalgene™, or comparable container, to provide water for the remainder of the sample processing. This 3-liter subsample is essentially filtration with the filtrate becoming the dissolved nutrient sample and the filters retained for chlorophyll a. Unfiltered water shall be taken for Total Suspended Solids (TSS) samples.

Approximately 40 milliliters (mL) of filtrate from the above chlorophyll filtration (surface water) shall be collected into a prelabeled, clean 60mL Nalgene™, or comparable container, screw-capped bottle, and stored in a freezer.

Before placing the sample in the freezer, record the approximate salinity (±2ppt (parts per thousand)) on the container. Depending on the analytical instrumentation used, matrix matching of solutions (e.g., standards or wash solutions) may be required for certain of the analytes. The nutrient samples shall remain frozen until they are to be analyzed.

2.3.2. Composited Surficial Sediment

multiple sediment grabs shall be taken by Van Veen or comparable sampler and the surficial sediment layer (top 2-3 centimeters (cm)) shall be collected by spatula or scoop and composited to provide sediment for the analyses of chemical contaminants, Total Organic Carbon (TOC)/Total Inorganic Carbon (TIC), toxicity testing, and grain size determinations. The number of grabs required to yield an adequate volume of composited sediment depends on the surface area described by the particular grab; however, surficial sediment from a minimum of three grabs shall be composited for the final sample. Surficial sediment from the individual grabs shall be combined into a clean, pre-labeled glass jar. Between grabs, the container of composited sediment shall be held on ice and covered with a lid to protect the sample from contamination (e.g., fuel or combustion products). Sediment samples shall be homogenized back in the laboratory, if not already done so in the field, prior to any analyses.

Organic chemical contaminants – approximately 500mL of composited sediment shall be placed into clean, pre-labeled, I-Chem™ jars, or comparable jars; fill containers to approximately 75% of capacity to allow for expansion during freezing. The sample shall be held on wet ice aboard the sampling vessel and, upon transfer to shore storage, the sample may be frozen or kept on ice for shipment back to the laboratory which shall be directed by the COR.

Trace element sediments shall be a 200mL subset of the field collected sample.

When samples are collected for toxicity testing – approximately 2000 – 4000mL (depending on the number of toxicity tests to be performed) of composited sediment shall be placed in clean, pre-labeled, jars. Specific volumes and container type(s) shall be dictated by the bioassays to be performed, ranging from 50mL glass jars to 2-4L plastic bottles. Prior to the field effort the COR shall specify which bioassays shall be conducted. The sample(s) shall be held on wet ice aboard and, upon transfer to shore storage, the sample shall be held at 4°C unless frozen, to await further processing and initiation of testing at the bioassay lab(s).

TOC – approximately 30mL of composited sediment shall be placed in a small, clean, pre-labeled glass bottle/jar.

The sample shall be held on wet ice aboard and upon transfer to shore storage, the sample may be frozen to await further laboratory analysis. The TOC sample may also be a subset of the 500mL sample taken for chemistry analyses.

Grain size determination – approximately 120mL of composited sediment shall be placed in a clean, pre-labeled, Whirlpak™ or comparable bag. The sample shall be held on wet ice in the field and, upon transfer to shore storage, the sample shall be held at 4°C (sample is not to be frozen) to await further laboratory processing.

2.3.3. Composited Bivalve Samples

At each site, bivalves are to be collected from three independent stations. Offshore subtidal sites are 400 meter (m) radius circles that the three stations per site shall be collected within. Shoreline intertidal sites are defined as a 100m linear distance along a tidal horizon to either side of the site center (local conditions may restrict this distance to less than 100m). These station samples shall be packaged separately and returned to the laboratory. Approximately 100 bivalves shall be collected from sites requiring only one trace element and organic analysis. The approximate number of bivalves to be collected may be changed by the MWP PM. For smaller mollusks such as zebra mussels it may require as many as 1,500 organisms to attain enough sample material such that all analyses can be completed.

All sites shall have bivalves of a specific species and size (mussels: 5-8cm for Mytilus sp. and 1.7-3cm for Dreissena polymorpha/D. bugensis; oysters: 7-10cm for Crassostrea virginica, 7-10cm for C. rhizophorae, and 1-2cm for

Ostrea sandvicensis; and 3-5cm for Chama sinuosa) that are available for collection annually. Additional species to be collected may be specified by the COR. Each site historically has had sufficient bivalves such that repeated biannual harvesting does not seriously deplete the resource, allowing for sampling in subsequent years (e.g., it is not anticipated that the site shall be physically disrupted by anthropogenic activities).

In the field and during shipping, bivalve samples are to be preserved on ice in a manner that shall keep the organisms alive. Bivalves can stay alive up to four days after collection if properly preserved. Use extra caution when ambient air temperatures drop below freezing; ensure iced samples do not get too cold. This is especially important for samples that shall be analyzed for histopathology/gonadal index.

The following data and information shall be collected by the field team: shell length, total weight of all bivalves collected, displacement volume with and without the soft tissues, percent wet and dry weight, lipid content, gonadal index, based on a separate sample of five or ten bivalves from each site, wet and dry weight of tissue analyzed, latitude, longitude, physical description of each sampling site including: digital color photographs, sites plotted on

NOAA charts or other mapping products (e.g., GIS, GoogleEarth™, etc.), directional information on how to access the sites, and date and time of collection.

2.3.4. Bioeffects and special study sampling

NCCOS reserves the right to contract sampling for Bioeffects and special studies. Bioeffects often requires sediment sampling, which would be executed in a manner consistent with section 2.3.2 above. NCCOS occasionally also requires sampling of other types of tissue (e.g., fish, coral, etc.). Fish may be sampled either as whole tissue, or pre-dissected (i.e., liver, etc.). Corals are typically collected as small cores and/or fragments. Detailed protocols are referenced in Section 5. REFERENCES (Apeti 2012a, Kimbrough 2006, and Kimbrough 2007) for these sampling efforts. The contractor shall obtain all relevant permits for collecting these samples.

2.4. Sample Handling and Shipping

Samples collected from the field may be temporarily held in the field, under appropriate conditions, for one to five days while awaiting shipment (or delivery) to centralized storage facilities or processing laboratories. Samples that shall be used for the determination of gonadal index and histopathology shall be shipped on ice (not frozen) as soon as possible after collection and shall not be held for more than three days. Sample handling and storage guidelines are presented in Table 2.

Table 2. Sample handling and storage guidelines for NS&T Monitoring.

Sample Type Container Field Holding Lab Storage Analysis

Sediment:

Organic contaminants Appropriately cleaned glass jars or I-Chem

Wet ice (4°C) or blude ice packs Freezer (-20°C) < 3 months

Inorganic contaminants 125-cc Nalgene Wet ice (4°C) or blude ice packs Freezer (-20°C) < 3 months

Total organic carbon Small glass jar Wet ice (4°C) or blude ice packs Freezer (-20°C) < 3 months

Silt/Clay 125-cc Nalgene Wet ice (4°C) or blude ice packs Refrigerator (4°C) < 3 months

Biota:

Tissue

Individual foil wrapped and combined in Ziploc bag

Wet ice (4°C) or blude ice packs Freezer (-20°C) < 3 months

Histopathology As per sample size Dietrich’s fixative*

Transfer to 70% ethanol 6 months

Biomarker/Genomic Individual in Teflon™ bag inside mesh bag Dry ice Freezer (-80°C) < 3 months

Passive Samplers:

Polar Organic Chemical

Integrated Sampler

(POCIS) /Polyethylene

Devices (PEDs)/

Semipermeable

Membrane Devices

(SPMDs) / Continuous

Low-level Aquatic

Monitoring (CLAM)s or similar

Appropriately cleaned glass/metal container

Wet ice (4°C) or blude ice packs Freezer (-20°C) < 3 months

*Fixative contains formalin which is a hazardous material and shall be shipped following all federally mandated hazardous material shipping procedures.

2.5. Documentation Requirements

A comprehensive project requires a structured system to ensure that all pertinent data are documented, and that samples are appropriately labeled, handled, stored, and transferred through all phases from field collection to final analysis. Although standard formats for data/sample collection and reporting shall be established for field and laboratory activities, not all aspects of sample handling shall be addressed by the forms alone. Therefore, additional written documentation is required to augment cradle to grave history for sample possession.

Field crews, whether NCCOS, contractors, or regional partners (i.e., state, academic, and federal personnel) shall record in-the-field data on hardcopy field sheets (see Appendix 6 for example) and, at a later date, all field data shall be transcribed into an electronic format for transmission. All field sheets shall be identified with a station

Identification (ID) code and dated; upon completion of the field entries, the person recording the data shall sign each sheet. Field sheets are designed to lead the sampling team through a logical sequence of steps and checks that further ensures sampling protocols are followed. The Field Lead shall verify that all field sheets are accounted for and complete prior to departing the sampling station.

All core data recorded on field sheets shall be transcribed into an electronic format within a week following collection (target period, within a week). The fields in the electronic format shall resemble the hardcopy raw data forms. The hardcopy data forms filled out for a given station shall be compiled into a “station data package” and supplied to NCCOS as Portable Document Format (PDF) files within a week of the collection of a particular site.

2.5.1. Chain of Custody

The contractor shall be able to accept delivery of field samples on every day of the week including weekends, and every effort shall be made to make deliveries during business hours on Monday through Friday.

NCCOS does not require the stringency of Good Laboratory Practices (GLPs) – Chain-of-Custody protocols, but the following level of accountability is expected. Field crews shall inventory samples and temporarily store them under designated conditions to await shipment or delivery to the processing laboratories. An example Chain-of-Custody form can be found in Appendix 5, and the Chain of Custody requirements for the field collection team and the receiving laboratory are as follows:

Field collection team –

1) A master inventory of all field samples that are expected to be collected (separate list(s) for each sample type and corresponding station IDs), with check off fields providing – documentation of all samples that are collected (when, and by whom)

2) Sample transfer information/invoice (where, what, to whom, and when, and by whom samples are transferred or shipped)

Laboratory receiving the samples –

1) Documentation (sample log-in form) of the person receiving; when and what they receive; and general condition of shipment (e.g., breakage, thawed, etc.)

2) Reconciliation that what was reported shipped was in fact received

3) Deposition/distribution of samples (e.g., where stored and holding conditions), with a master electronic list and a hardcopy list filed in the general area where the samples are held.

4) Check-out/check-in procedure for sample release to analysts and subsequent return to storage.

The field team shall retain copies of shipping invoices and the originals shall be sent with the samples as they are transferred. The field copies shall be compiled into a complete set and submitted to NCCOS (as PDFs) for archiving.

The recipient of the samples (processing laboratory) shall inventory the physical samples against the invoice and alert NCCOS in the event of any missing samples. If a sample is missing, the laboratory shall go through appropriate channels to contact the field team as soon as possible so that they may attempt to locate the sample at their end or possibly re-sample.

3. LABORATORY ANALYSES

Contractors shall ensure that all NCCOS samples analyzed meet the protocols, standards, and quality guidelines presented herein.

3.1. Sample Disposition

The receiving lab participates in the transfer documentation as outlined in section 2.5 above. Samples shall be retained by the laboratory for each site sampled, for each year samples were collected for at least five years. These samples shall be reanalyzed, at no addition cost, should data problems arise that cannot be resolved by other means.

If necessary, samples shall be sent to a referee laboratory for confirmation analysis.

Left over samples (bivalve and sediments), sample homogenates, and sample extracts are the property of NCCOS, but are to be kept by the contract labs for five years unless requested by NCCOS. Samples kept for more than five years may be shipped to NCCOS, at no additional cost, at the end of the contract period.

3.2. Analytical Methods Requirements

Analytical procedures for NS&T range from straightforward determinations such as percent gravel, sand, silt, and clay to comprehensive analyses of chemical contaminants in complex environmental matrices. Examples of these procedures for the various analyses are described in Kimbrough and Lauenstein (2006) and Kimbrough et al. (2007).

3.2.1. Chemical Contaminants

The laboratory analyses of NS&T samples may include analyses of sediment, fish, bivalve, coral, and water samples along with passive sampling media (POCIS, PEDs, etc.). All NS&T studies, whether Mussel Watch, Bioeffects, or

Special Studies, require contaminant chemistry measurements. Past methods used to perform these analyses are documented in NOAA Technical Memoranda that are detailed in the reference section below (section 5).

The contractor shall address the suite of core and intermittent NS&T analytes listed in Table 3. Core analytes includes toxic and trace elements and legacy organic contaminants (such as PAHs, PCBs) while intermittent contaminants are those analytes that are measured infrequently. The contaminants listed in Table 3 do not represent a comprehensive list: other contaminants including currently unidentified compounds may be requested at a later date as they become environmentally relevant. Pricing for newly requested contaminants shall be discussed at the time of request, and the inability of the contractor to successfully conduct the newly requested analysis after a reasonable amount of attempts may not be held against the contractor.

No specific methods are required for these analyses; it is left to each laboratory to develop its own best analytical procedures based on their available resources, personnel and bench state of the art. However, the QA/Quality

Control (QC) for these analyses is performance-based, and the laboratory shall meet the minimum quality criteria set forth. The NCCOS performance-based approach to QA/QC for analytical chemistry is involved and is described in detail in Kimbrough et al. (2006 and 2007). Although each laboratory develops their own method for most of the analyses to be performed, certain approaches are generally accepted. The contractor shall refer to NS&T method documents listed in section 5. Methods shall be cited or presented for all contaminants.

Table 3. Required contaminant analyses. This is not a comprehensive list of required analytes, other contaminants including currently unidentified compounds shall be requested as they become environmentally relevant.

Routine analyses of NS&T samples shall be conducted in batch runs consisting of 20 or fewer field samples along with a full complement of QC samples, typically including: continuing calibration curves, reagent blanks, Matrix

Spikes (MS) and MS Duplicates (MSD), and a Reference Material (RM either a Standard RM (SRM) or a laboratory control material). Refer to Kimbrough et al. (2006 and 2007) for a comprehensive discussion of the performance-based QC philosophy and components. If the quality criteria are not met, the analyst shall take corrective actions and rerun the batch. When laboratories adhere to this level of in-house data review, only batches that pass the general

QC checks shall be submitted as final data to NCCOS.

When these QC checks are embedded in each batch, the analyst shall assess the overall data quality on a per batch basis and shall take corrective measures if there are deficiencies. If data for a class of compounds consistently fails any of the NS&T quality standards, the laboratory management shall notify NCCOS of the problem and seek recommended corrective action prior to submitting the final data report. Kimbrough et al. (2006 and 2007) present a comprehensive listing of the key quality control elements for chemical analysis and are available online at:

https://coastalscience.noaa.gov/.

3.2.2. Contaminants of Emerging Concern (CECs)

CECs represent a broad class of both chemicals and the associated analytical methods to quantify them. By their nature, analysis of this type of chemical currently represents intermittent work within the scope of this SOW.

However, during the course of this contract the nature and importance of individual CECs may change and represent a larger requirement than currently proposed.

Current CECs that are of specific interest to NS&T include, but are not limited to: Human Use Pharmaceuticals, Personal Care Products, hormones, nonyl-phenols, chlorinated paraffins, and microplastics. Due to the complex nature of, and continually developing technologies and methods for measuring CECs, pricing for CECs can be discussed on a per request basis, though price increases of greater than 10% above the initial discussed price shall require written justification be provided to the COR.

3.2.3. Biotic Health Indicators

Biotic health indicators also represent a broad class of laboratory analysis. Analysis of this type represents intermittent work within the scope of this SOW. As with CECs, during the course of this contract the nature and importance of individual biotic health indicators may change and represent a larger requirement than currently proposed.

The biotic health indicators that are of specific interest include, but are not limited to: metabolomics, transcriptomics, proteomics, lipidomics, genomics, and cellular biomarkers (e.g., GSH, Lipox, AcHE). Due to the complex nature of, and continually developing technologies and methods for measuring biotic health indicators, pricing for these types of analysis can be discussed on a per request basis, though price increases of greater than

10% above the initial discussed price shall require written justification be provided to COR.

3.2.4. Water Quality Parameters

When requested by NCCOS, water quality shall be evaluated for the analyses of indicators of anthropogenic enrichment, including nutrient levels. Samples for these indicators shall be obtained by filtering site water (collected https://coastalscience.noaa.gov/ at the depth regimes described in Section 2.3.1) and retaining the material filtered out for subsequent analyses if requested; the filtrate shall be used for the analyses of soluble nutrients.

Currently nitrate and nitrite analyses are based on the methodology of Armstrong et al. (1967) and utilize cadmium for reduction of NO3

- to NO2

-. Orthophosphate is measured using chemistry based on the investigations of Bernhardt and Wilhelms (1967) with the modification of hydrazine as a reductant. Silicate determination is accomplished using the methods of Armstrong et al. (1967) using stannous chloride. Ammonium analysis is based on the method of

Harwood and Kuhn (1970) using dichloro-isocyanurate as an oxidizer. Urea is measured using diacetyl-monoximine and themicarbozide. The total concentrations of nitrogen and phosphorus are determined after an initial decomposition step. This method involves persulfate oxidation while heating the sample in an autoclave (115 C, 20 minutes) (Hansen and Koroleff 1999). After oxidation of the samples, nutrient concentration is determined as nitrate and orthophosphate. Method Detection Limits (MDLs) for nutrient related analysis are found in Table 4.

Table 4. MDLs for nutrient related analysis.

Analyte Method Detection

Limit (µM)

Method Detection

Limit (mg/L)

Standard Range

(µM)

Standard Range

(mg/L)

NO3

- 0.177 0.01 3.85 - 30.14 0.23 - 1.86

NO2

- 0.01 0.0004 0.09 - 0.72 0.006 - 0.033

HPO4

- 0.03 0.002 0.35 - 2.18 0.021 - 0.21

HSIO3

- 0.155 0.014 4.05 - 30.08 0.25 - 2.80

NH4

+ 0.07 0.001 0.42 - 3.44 0.026 - 0.062

Urea 0.205 0.012 0.59 - 4.42 0.036 - 0.265

3.2.5. Sediment Particle Size

Particle size distribution is determined in marine sediments on the basis of the Wentworth scale method. The major size classes determined are; gravel (-2 phi to -5 phi), sand (+4 phi to -1 phi), silt (+5 phi to +7 phi), and clay (+8 phi and smaller). Determining particle size in sediments is important due to potential correlations with contaminant levels.

3.2.6. Total Organic Carbon

Analysis of sediment TOC shall be conducted for Mussel Watch and Bioeffects samples. Determining TOC in sediments is important due to potential correlations with contaminant levels.

3.2.7. Histopathology

Because the relative proportion of gonadal (20% to 50% of body weight) to somatic tissue can significantly impact the body burden of contaminants and because pollutants, pathology, and parasite body burdens in bivalves can be linked, the MWP began surveying the sentinel organisms for their reproductive state and histopathology parameters.

Assessment of the reproductive state of bivalve populations requires an analysis of the state of gonadal development.

Determination of reproductive stage is included as part of the MWP to give an indication of the amount of gametic material in bivalve tissues at the time of chemical analysis. The histopathology parameters measured in the bivalves include an array of parasites (Table 7a) such as prokaryotes (e.g., rickettsia, chlamydia), tapeworms (e.g., cestodes), and roundworms (e.g., nematodes). Oyster diseases and pathological conditions such as tumor, neoplasm and necrosis are also measured.

Bivalve reproductive state and histopathological condition shall be analyzed based on the SOP described in the

NOAA Technical Memorandum NOS NCCOS 27 (Kim et al. 2006). Though that reference allows a range of 5-15 animals for gonadal and histopathological analyses, subsequent data analysis indicates five replicates may be insufficient to fully characterize their prevalence. Therefore, NCCOS requires analysis of 12 individual bivalves for reproductive stage and 10 organisms for histopathology.

For the reproductive state characterization, the stage in the gametogenic cycle shall be assigned based on the maturity of the follicles and gametes, and a numerical value shall be assigned as described in the Tables 5 and 6.

Table 5. Mussel and Zebra Mussel Development Stages.

Reproductive stage Description

Resting/spent gonad

Stage 0 Inactive or undifferentiated

Developing gonad

Stage 1 Gametogenesis has begun; no ripe gametes visible

Stage 2 Ripe gametes present; gonad developed to about one-third of its final size

Stage 3 Gonad increased in mass to about half the fully ripe condition; each follicle contains, in area, about equal proportions of ripe and developing gametes

Stage 4 Gametogenesis still progressing, follicles contain mainly ripe gametes

Ripe gonad

Stage 5 Gonad fully ripe, early stages of gametogenesis rare; follicles distended with ripe gametes; ova compacted into polygonal configurations; sperm with visible tails

Spawning gonad

Stage 4 Active emission has begun; sperm density reduced; ova rounded off as pressure within follicles is reduced

Stage 3 Gonad about half empty

Stage 2 Gonadal area reduced; follicles about one-third full of ripe gametes

Stage 1 Only residual gametes remain; some may be undergoing cytolysis

Table 6. Oyster Development Stages

Developmental Stage Value Description

Sexually undifferentiated 1 Little or no gonadal tissue visible

Early development 2 Follicles beginning to expand

Mid-development 3 Follicles expanded and beginning to coalesce; no mature gametes present

Late development 4 Follicles greatly expanded and coalesced, but considerable connective tissue remaining; some mature gametes present

Fully developed 5 Most gametes mature; little connective tissue remaining

Spawning 6 Gametes visible in gonoducts

Spawned 7 Reduced number of gametes; some mature gametes still remaining evidence of renewed reproductive activity

Spawned 8 Few or no gametes visible; gonadal tissue atrophying

Tables 7a and 7b provide the list of histopathology parameters currently measured by the MWP. This is not by any means an exhaustive list of parasites and disease that can be found in bivalves; other relevant parameters should be augmented with this list. The histopathological examinations are based on quantitative or semi-quantitative approaches (Table 8). The quantitative approach is based on direct counts of parasites and pathologies, while the semi-quantitative evaluations use predetermined infection intensity scales.

Table 7a. List of parasites measured by the MWP as part of the histopathology assessment of bivalves.

Parasite Category Parasites

Cestodes Body cestodes, Gill cestodes, Mantle cestodes

Copepods Body Copepods, Gill Copepods, Gut Copepods

Ciliates Digestive tract Ciliates, Large gill Ciliates, Small gill Ciliates, Gut Ciliates

Protozoans Digestive tubule protozoan, Gut protozoan

Nematodes Nematode

Trematodes Trematode sporocyst, Trematode metacercaria, Proctoeces

Gregarines Body Gregarines, Gill Gregarines, Mantle Gregarines

Prokaryotes Digestive tubule rickettsia, Gut rickettsia, Chlamydia

Coccidians Pseudoklossia

Pea crabs Pinnotherid crab

Table 7b. List of diseases and tissue conditions/pathologies measured by the MWP as part of the histopathology assessment of bivalves.

Disease/tissue condition category Parasites/Tissue Conditions

Dermo P. marinus

MSX H. nelsoni

Digestive tubule conditions Digestive gland atrophy, Unusual digestive tubule

Edema Edema

Necrosis Focal necrosis, Diffuse necrosis

Neoplasia Neoplasm

Hemocytic infiltration Focal hemocytic infiltration, Diffuse hemocytic infiltration

Tumor Tumor

Xenoma Xenoma

Table 8. List of quantitative and semi-quantitative categories for each bivalve taxon.