Crofton Environmental Effects Monitoring (EEM) Cycle Five Interpretive Report
March 2010
Prepared for: Catalyst Paper Corporation Crofton, British Columbia
Suite 200 – 850 Harbourside Drive, North Vancouver, British Columbia, Canada V7P 0A3 • Tel: 1.604.926.3261 • Fax: 1.604.926.5389 • www.hatfi eldgroup.com CROFTON ENVIRONMENTAL EFFECTS MONITORING (EEM) CYCLE FIVE INTERPRETIVE REPORT
Prepared for:
CATALYST PAPER CORPORATION CROFTON DIVISION PO BOX 70 CROFTON, BC V0R 1R0
Prepared by:
HATFIELD CONSULTANTS #200 - 850 HARBOURSIDE DRIVE NORTH VANCOUVER, BC V7P 0A3
MARCH 2010
CR1327.2
#200 - 850 Harbourside Drive, North Vancouver, BC, Canada V7P 0A3 • Tel: 1.604.926.3261 • Toll Free: 1.866.926.3261 • Fax: 1.604.926.5389 • www.hatfieldgroup.com TABLE OF CONTENTS
LIST OF FIGURES ...... v LIST OF APPENDICES ...... viii ACKNOWLEDGEMENTS ...... ix EXECUTIVE SUMMARY ...... x
1.0 INTRODUCTION ...... 1-1
2.0 MILL, STUDY AREA, AND CYCLE FIVE DESIGN UPDATE ...... 2-1 2.1 MILL OPERATIONS ...... 2-1 2.1.1 Process Description and Update ...... 2-1 2.1.2 Effluent Quality ...... 2-4 2.1.3 Spills to the Receiving Environment ...... 2-5 2.1.4 Study Area Updates ...... 2-5 2.2 CYCLE FIVE STUDY DESIGN UPDATE ...... 2-5
3.0 SUBLETHAL TOXICITY TESTING OF MILL EFFLUENT ...... 3-1 3.1 SUBLETHAL TOXICITY TEST METHODS ...... 3-2 3.1.1 General Methods and Definitions ...... 3-2 3.1.2 Sublethal Toxicity Test Methods ...... 3-3 3.2 RESULTS AND DISCUSSION ...... 3-4 3.2.1 Topsmelt Growth and Survival Test ...... 3-4 3.2.2 Echinoderm Fertilization Test ...... 3-5 3.2.3 Algal Reproduction Test ...... 3-6 3.2.4 Potential Zone of Sublethal Effect ...... 3-7 3.3 CONCLUSIONS ...... 3-8
4.0 FISH POPULATION SURVEY ...... 4-1 4.1 INTRODUCTION ...... 4-1 4.2 METHODS ...... 4-2 4.2.1 Modifications to Sampling Design ...... 4-2 4.2.2 Control/Impact Design ...... 4-2 4.2.3 Sampling Locations and Collection Dates ...... 4-3 4.2.4 Field Sampling Procedures ...... 4-5 4.2.5 Laboratory Procedures ...... 4-6 4.2.6 Analytical Approach ...... 4-8 4.3 RESULTS ...... 4-14 4.3.1 Mussels ...... 4-14 4.3.2 Oysters ...... 4-25 4.3.3 Supporting Environmental Variables ...... 4-35 4.4 DISCUSSION ...... 4-38
Crofton EEM Cycle Five i Hatfield 5.0 FISH TISSUE SURVEY ...... 5-1 5.1 TISSUE ANALYSES: DIOXINS AND FURANS ...... 5-1 5.1.1 Sediment ...... 5-1 5.1.2 Crab Hepatopancreas ...... 5-1 5.2 TAINTING EVALUATION ...... 5-3
6.0 BENTHIC INVERTEBRATE COMMUNITY SURVEY ...... 6-1 6.1 INTRODUCTION ...... 6-1 6.2 METHODS ...... 6-2 6.2.1 Modifications to Sampling Design ...... 6-2 6.2.2 Gradient Design ...... 6-2 6.2.3 Sampling Locations and Collection Dates ...... 6-2 6.2.4 Field Sampling Procedures ...... 6-3 6.2.5 Field Processing Procedures ...... 6-7 6.2.6 Laboratory Procedures ...... 6-8 6.2.7 Analytical Approach ...... 6-9 6.3 RESULTS ...... 6-13 6.3.1 Benthic Invertebrate Communities ...... 6-13 6.3.2 QA/QC and Verifications ...... 6-24 6.3.3 Supporting Environmental Variables ...... 6-24 6.4 DISCUSSION ...... 6-36 6.4.1 Effects Along the Exposure Gradient ...... 6-36 6.4.2 Grades of Impact Along the Exposure Gradient ...... 6-38 6.4.3 Characterizing Cycle Five Effects ...... 6-41
7.0 CONCLUSIONS ...... 7-1
8.0 CLOSURE ...... 8-1
9.0 REFERENCES ...... 9-1
10.0 GLOSSARY ...... 10-1
Crofton EEM Cycle Five ii Hatfield LIST OF TABLES
Table 2.1 Annual results for process effluent quality variables, Catalyst Paper, Crofton Division, 2007 to 2009...... 2-3 Table 3.1 Potential Zone of Sublethal Effect, Catalyst Paper Corporation, Crofton Division, EEM Cycle Five...... 3-8 Table 4.1 Bivalve survey control/impact sampling design, Crofton EEM Cycle Five, 2009...... 4-2 Table 4.2 Bivalve survey sampling locations and collections, Crofton EEM Cycle Five, 2009...... 4-3 Table 4.3 Statistical analyses performed to evaluate effects on bivalves, Crofton EEM Cycle Five, 2009...... 4-13 Table 4.4 Whole-organism metric summary statistics for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-16 Table 4.5 Whole-organism indices summary statistics for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-17 Table 4.6 Tests for differences in mussel whole organism metrics and indices among near-field, far-field, and reference (far-far-field) stations, Crofton EEM Cycle Five, March 2009...... 4-22 Table 4.7 Response-based effects assessment for mussels using statistical results, Crofton EEM Cycle Five, March 2009...... 4-22 Table 4.8 Comparison of Post hoc power analysis of ANCOVAs conducted to test for differences in mussel condition and GSI, and a priori analysis of number of samples actually required to achieve sufficient power, Crofton EEM Cycle Five, March 2009...... 4-24 Table 4.9 Whole-organism metric summary statistics for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-26 Table 4.10 Whole-organism indices summary statistics for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-27 Table 4.11 Tests for differences in oyster whole organism metrics and indices among near-field, far-field, and reference (far-far-field) stations, Crofton EEM Cycle Five, July 2009...... 4-33 Table 4.12 Response-based effects assessment for oysters using statistical results, Crofton EEM Cycle Five, July 2009...... 4-33 Table 4.13 Results of post hoc power analysis of ANCOVAs conducted to test for differences in oyster condition, Crofton EEM Cycle Five, July 2009...... 4-35 Table 4.14 Habitat characteristics and water quality at Pacific blue mussel sampling stations, Crofton EEM Cycle Five, March 2009...... 4-36
Crofton EEM Cycle Five iii Hatfield Table 4.15 Habitat characteristics and water quality at Pacific oyster sampling stations, Crofton EEM Cycle Five, July 2009...... 4-36 Table 6.1 Benthic invertebrate sampling locations and collections, Crofton EEM Cycle Five, 2009...... 6-4 Table 6.2 Summary of benthic invertebrate community statistics, Crofton EEM Cycle Five, March 2009...... 6-15 Table 6.3 Forty most abundant benthic taxa, Crofton EEM Cycle Five, March 2009.1 ...... 6-16 Table 6.4 Results of regressions and correlations conducted to test relationships between benthic community variables and gradients of pulpmill effluent exposure, Crofton EEM Cycle Five, March 2009...... 6-20
Table 6.5 Results of Spearman rank correlations (rs) among all benthic invertebrate metrics (n=12), Crofton EEM Cycle Five, 2009...... 6-22 Table 6.6 Habitat characteristics, near-bottom water quality, and sediment quality at benthic invertebrate community survey stations, Crofton EEM Cycle Five, 2009...... 6-27 Table 6.7 Results of regression analyses conducted to test relationships between supporting environmental variables and gradients of pulpmill effluent exposure (distance from the outfalls and C:N), Crofton EEM Cycle Five, 2009...... 6-32
Table 6.8 Results of Spearman rank correlations (rs) among sediment and near-bottom water quality variables (n=12), Crofton EEM Cycle Five, 2009...... 6-34
Table 6.9 Results of Spearman rank correlations (rs) between benthic community metrics and supporting variables (n=12), Crofton EEM Cycle Five, 2009...... 6-35 Table 6.10 Summary of observed responses of benthic invertebrate communities with C:N ratio exposure gradient, Crofton EEM Cycle Five, March 2009...... 6-37 Table 6.11 Summary of observed responses of benthic invertebrate communities with distance from outfalls, Crofton EEM Cycle Five, March 2009...... 6-37 Table 6.12 Scheme used to grade impacts of sediment chemistry on benthos...... 6-39 Table 6.13 Assessment of sediment quality and benthic community health at each station sampled for Crofton EEM Cycle Three, February 2003, based on “impact grades” provided in EEM Technical Guidance.1 ...... 6-40 Table 6.14 Assessment of sediment quality and benthic community health at each station sampled for Crofton EEM Cycle Five, March 2009, based on “impact grades” provided in EEM Technical Guidance.1 ...... 6-41
Crofton EEM Cycle Five iv Hatfield LIST OF FIGURES
Figure 2.1 Location of Catalyst Paper, Crofton Division, on Stuart Channel, British Columbia...... 2-2
Figure 2.2 Annual production and effluent flows from 1970 to 2009 Catalyst Paper Corporation, Crofton Division...... 2-4
Figure 2.3 Mean daily total suspended solids (TSS), biochemical oxygen demand (BOD), and adsorbable organic halides (AOX) in effluent, Catalyst Paper Corporation, Crofton Division, 1981 to 2009...... 2-6
Figure 3.1 Effect of exposure to Catalyst Paper, Crofton Division effluent on topsmelt survival, expressed as LC50 ±95% confidence limits, EEM Cycle Five...... 3-4
Figure 3.2 Effect of exposure to Catalyst Paper, Crofton Division effluent on topsmelt growth, expressed as IC25 ±95% confidence limits, EEM Cycle Five...... 3-5
Figure 3.3 Effect of exposure to Catalyst Paper, Crofton Division effluent on echinoderm fertilization, expressed as IC25 ±95% confidence limits, EEM Cycle Five...... 3-6
Figure 3.4 Effect of exposure to Catalyst Paper, Crofton Division effluent on algal reproduction, expressed as IC25 ±95% confidence limits, EEM Cycle Five...... 3-7
Figure 3.5 Geometric means of IC25 and LC50 results from sublethal toxicity tests of Catalyst Paper, Crofton Division effluent for EEM Cycle One through Cycle Five...... 3-8
Figure 4.1 Pacific oyster and mussel sampling locations, Crofton EEM Cycle Five, 2009...... 4-4
Figure 4.2 Mean length, width and girth (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-15
Figure 4.3 Mean whole wet weight (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-15
Figure 4.4 Weight distributions for Pacific blue mussels by station, Crofton EEM Cycle Five, March 2009...... 4-17
Figure 4.5 Mean condition (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-18
Figure 4.6 Mean shell density (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-18
Figure 4.7 Sex ratios for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-19
Crofton EEM Cycle Five v Hatfield Figure 4.8 Mean GSI (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-19
Figure 4.9 Mean age (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-20
Figure 4.10 Mean size-at-age (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009...... 4-20
Figure 4.11 Age distributions for Pacific blue mussels by station, Crofton EEM Cycle Five, March 2009...... 4-21
Figure 4.12 Oyster density at stations sampled for Crofton EEM Cycle Five, July 2009...... 4-25
Figure 4.13 Mean length, width and girth (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-28
Figure 4.14 Mean whole wet weight (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-28
Figure 4.15 Weight distributions for Pacific oysters by station, Crofton EEM Cycle Five, July 2009...... 4-29
Figure 4.16 Mean condition (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-29
Figure 4.17 Mean shell density (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-30
Figure 4.18 Mean age (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-31
Figure 4.19 Age distributions for Pacific oysters by station, Crofton EEM Cycle Five, July 2009...... 4-31
Figure 4.20 Mean size-at-age (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009...... 4-32
Figure 4.21 Near-shore water temperatures at bivalve sampling stations, Crofton EEM Cycle Five, March to July 2009...... 4-37
Figure 5.1 Estimated dispersion of effluent from Catalyst Paper, Crofton Division, and 2009 fisheries closures in Stuart Channel...... 5-2
Figure 5.2 Crofton sediment 2,3,7,8-T4CDD, T4CDF and TEQ, 1990 to 2009...... 5-4
Figure 5.3 Crofton Dungeness crab hepatopancreas 2,3,7,8-T4CDD, 1990 to 2009...... 5-5
Figure 5.4 Crofton Dungeness crab hepatopancreas 2,3,7,8-T4CDF, 1990 to 2009 ...... 5-7
Crofton EEM Cycle Five vi Hatfield Figure 5.5 Crofton Dungeness crab hepatopancreas 2,3,7,8-T4CDD TEQs (WHO 2005 TEFs), 1990 to 2009...... 5-9
Figure 6.1 Crofton EEM Cycle Five benthic invertebrate sampling locations, March 2009...... 6-5
Figure 6.2 Mean adult benthic invertebrate density (± SE) per station, Crofton EEM Cycle Five, March 2009...... 6-14
Figure 6.3 Taxonomic richness (total per station) of benthic invertebrate communities, Crofton EEM Cycle Five, March 2009...... 6-17
Figure 6.4 Evenness of benthic invertebrate communities, Crofton EEM Cycle Five, March 2009...... 6-17
Figure 6.5 Simpson's Diversity across benthic invertebrate communities, Crofton EEM Cycle Five, March 2009...... 6-18
Figure 6.6 Bray-curtis similarity indices for benthic communities, Crofton EEM Cycle Five, March 2009...... 6-19
Figure 6.7 Significant regressions of mean adult density, Bray-Curtis Index, and evenness against C:N ratio and distance from outfalls, showing 95% confidence intervals, Crofton EEM Cycle Five, March 2009...... 6-21
Figure 6.8 Significant Spearman rank correlations of benthic metrics, Crofton EEM Cycle Five, 2009...... 6-22
Figure 6.9 Dendrogram describing similarity of benthic invertebrate communities at station sampled for Crofton EEM Cycle Five, based on Bray-Curtis dissimilarity coefficients among stations...... 6-23
Figure 6.10 Particle size1 distribution of sediments at benthic invertebrate community survey stations, Crofton EEM Cycle Five, 2009...... 6-25
Figure 6.11 Near-bottom water quality at benthic invertebrate community survey stations, Crofton EEM Cycle Five, 2009...... 6-26
Figure 6.12 TOC, TN, and C:N ratio at benthic invertebrate community survey stations, Crofton EEM Cycle Three (2003) and Cycle Five (2009)...... 6-30
Figure 6.13 Redox potential and sulphides at benthic invertebrate community survey stations, Crofton EEM Cycle Three (2003) and Cycle Five (2009)...... 6-31
Figure 6.14 Scatterplots showing significant Spearman rank correlations (rs) and best-fit regression line between benthic community metrics and supporting variables, Crofton EEM Cycle Five, 2009...... 6-36
Crofton EEM Cycle Five vii Hatfield LIST OF APPENDICES
Appendix A1 Sublethal Toxicity Data and Calculations
Appendix A2 Bivalve Survey: ALS Tissue Analytical Reports
Appendix A3 Bivalve Survey: Metrics and Indices Data
Appendix A4 Tissue Survey: AXYS Sediment and Crab Analytical Reports
Appendix A5 Benthic Survey: ALS Sediment Analytical Reports
Appendix A6 Benthic Survey: Aquametrix Analytical Reports
Appendix A7 Benthic Survey: Columbia Science Invertebrate Data and QA/QC Reports
Appendix A8 Benthic Survey: Power Analysis for Regressions
Appendix A9 Benthic Survey: Redox Potential and Sulphides Preparation, Calibration, and Analysis
Crofton EEM Cycle Five viii Hatfield ACKNOWLEDGEMENTS
Primary investigators for the EEM Cycle Five program for Catalyst Paper, Crofton Division, from Hatfield Consultants included Nara Henderson, Noah Baker, and Martin Davies. Susan Stanley prepared the maps, and Christina Delapaz assisted with report production.
Thanks are due to the following people who assisted with field collections:
Mike Hale (crabbing skiff Captain) and his assistant Alex; Nathan Blasco (MV Lobo Captain, Aquametrix); and Dave Stirling (MV Lobo Onboard Redox/Sulphides Chemist, Aquametrix).
Noah Baker and Dan Moats (Environmental Specialists, Hatfield) assisted with laboratory tissue dissections.
We are also grateful to the following analytical subcontractors involved in the project:
ALS Environmental (Vancouver, BC); AXYS Analytical Services Ltd. (Sidney, BC); Cantest (Vancouver, BC); and Dawna Brand (UVic Biology).
The Crofton EEM Local Monitoring Committee (LMC) includes representatives from the federal and provincial governments, non-governmental organizations, community members, First Nations, Hatfield Consultants, and Catalyst Paper, Crofton Division. LMC meetings provided a valuable forum for reviewing results from the previous EEM Cycles, and discussing the design for the Cycle Five program. Hatfield would like to acknowledge members of the Crofton LMC for their assistance:
Janice Boyd (Environment Canada); Bob More (Environment Canada); Rosie Barlak (BC Ministry of Environment); Bernard Bintner (BC Ministry of Environment); Vern White (MP, Office of Nanaimo-Cowichan); Carol Donnelly (CC Advisory Group); Andrew McNaughton (Halalt); John. T. Elliott (Cowichan Tribes Fisheries); Drew Kilback (Catalyst Paper); Michelle Vessey (Catalyst Paper, Crofton Division); Darlene Walkey (Catalyst Paper, Crofton Division); and Bob Ericksen (Catalyst Paper, Crofton Division).
Crofton EEM Cycle Five ix Hatfield EXECUTIVE SUMMARY
The Environmental Effects Monitoring (EEM) Cycle Five program for Catalyst Paper, Crofton Division ran between April 2007 and April 2010, and included process effluent sublethal toxicological testing, and fish population, fish tissue, and benthic invertebrate community surveys.
Sublethal toxicity testing during Cycle Five demonstrated no effects of mill effluent on survival or growth of topsmelt (Atherinops affinis) larvae, effects on echinoderm fertilization at a mean effluent concentration of 23% (IC25), and effects on Champia parvula reproduction at a mean effluent concentration of 1.55% (IC25). Based on a 1% effluent concentration zone of 600 m from the outfall, maximum potential zones of sublethal effect from the effluent discharge point were <6 m for fish survival, 26 m for invertebrate fertilization, and 388 m for algal reproduction.
Dioxin and furan concentrations in sediments and crab near the Crofton mill have dropped considerably following pollution abatement changes in the 1980s and 1990s, including the gradual phase out of elemental chlorine bleaching and its complete elimination in 1996. Although the rate of decline has since slowed, concentrations continue to drop and have not yet leveled off. In 2009, Total Toxic Equivalency (TEQ) concentrations in crabs at four stations were below Health Canada’s consumption threshold (24.4 pg/g hepatopancreas), and levels at the remaining three stations had declined relative to 2006.
A fish population survey was conducted using bivalves to determine the effects of effluent exposure on fish populations near the Crofton mill. The study objective was achieved by sampling mussels and oysters at three stations: CRO3A (near-field), CRO5A (far-field), and CRO4 (far-far-field reference). The control/impact design was used to determine if significant differences (i.e., effects resulting from effluent exposure) could be detected.
Lower densities of older, larger, heavier mussels were present closer to the mill, indicating greater survival, energy use, and energy storage in these organisms relative to those at the reference station. In contrast, oysters in the near-field demonstrated lower survival, energy use, and energy storage than reference oysters; near-field oysters were significantly smaller and lighter, both in general and for their age, and were present in higher densities relative to other stations.
Ratios of male versus female mussels were similar between the near-field and reference areas, and similar GSI between the two areas indicated that sex did not play a role in influencing the significantly greater survival, energy use, and energy storage of mussels in the near-field. Mussels were older in the near-field, indicating that recent spawning and/or spat settlement was not as successful at the near-field station. In contrast, the youngest oysters in the study were only found at the near-field station, and in terms of their size-at-age the near-field oysters were substantially smaller.
Crofton EEM Cycle Five x Hatfield Coexistence of mussels and oysters can be difficult, with the larger oysters often colonizing suitable habitat at the expense of mussels; given the more favourable growing conditions observed for oysters at the reference station (i.e., higher temperature and DO), and the presence of larger oysters and smaller mussels at this station, it is possible that oyster populations have settled at the reference station at the expense of mussels.
In contrast to previous cycles, the response-based effects summary for Cycle Five demonstrated significant effects between the exposure and reference areas for both mussels and oysters. Given that opposing effects were observed for the two species, it is difficult to say whether the differences are associated with natural or mill-related factors.
A benthic invertebrate community survey was conducted to determine effects of effluent exposure on benthos in the Crofton area. The survey objective was achieved by sampling for benthic invertebrates, supporting sediment and water quality data, along a gradient of twelve stations placed at increasing distances from the mill outfalls. The gradient design was based on those performed in Cycles Two and Three, and made it possible to determine if effects on benthic invertebrates were evident along the exposure gradient.
Although sediment quality has improved since Cycle Three, closer to the outfalls sediments continued to be characterized by higher C:N, organic carbon, and lower redox potential. Although conditions have improved since Cycle Three, benthic communities in Cycle Five continued to demonstrate effects along the effluent exposure gradient. Closer to the outfalls, benthic communities exhibited higher densities, were dominated by Capitellid worms, and were statistically less similar to reference communities farther away.
The impact grading scheme recommended for use by federal guidance was applied to the results, and supported the conclusion that conditions since Cycle Three have improved; Cycle Five near-field sediment quality ranged between normal to moderately impacted, and benthic communities were representative of normal to low impact/enrichment conditions. As in Cycle Three, the Cycle Five benthic study demonstrated that, although the historical fibre mat in the vicinity of the Crofton outfalls has decomposed to a sufficient extent that it no longer exerts a toxic (inhibitory) effect on benthic communities, the remaining organic matter continues to contribute to a mild enrichment effect at stations close to the outfalls.
Crofton EEM Cycle Five xi Hatfield 1.0 INTRODUCTION
Under the federal Pulp and Paper Effluent Regulations (PPER; originally released in 1992, revised in May 2004 (Government of Canada 2005), and amended in 2008) pulpmills are required to monitor the chemistry and toxicity of mill effluent and assess its potential effects on the receiving environment. Effluent chemistry (limited to total suspended solids and biological oxygen demand) and acute toxicity are measured to evaluate effluent quality and its potential effects on aquatic biota. However, because many factors can alter the chemistry and toxicity of effluent in the receiving environment, Environmental Effects Monitoring (EEM) studies also are conducted to directly assess the effects of mill effluent on fish, fish habitat, and use of fisheries resources in the vicinity of the effluent discharge (Environment Canada 2005). EEM studies can include:
A fish population survey to assess fish health;
A fish tissue survey to assess concentrations of dioxins and furans (only required for mills where dioxins and furans are present in mill effluent, or where biotic concentrations in the receiving environment exceed Health Canada thresholds);
A benthic invertebrate community survey to assess the condition of fish habitat;
Supporting water quality data to help interpret findings from fish and benthic invertebrate surveys; and
Sublethal toxicity testing to assess effects of effluent on growth and reproduction of representative aquatic organisms.
EEM programs typically are conducted in three year cycles, which begin with the development of a study design, followed by study implementation, data analysis, and reporting. The following cycles have been completed for the Crofton Mill since the onset of the monitoring program:
Cycle One: 1993 to 1996;
Cycle Two: 1997 to 2000;
Cycle Three: 2000 to 2004; and
Cycle Four: 2004 to 2007.
The current program, Cycle Five, ran between April 2007 and April 2010. All components of the EEM program are conducted in accordance with the Pulp and Paper Technical Guidance for Aquatic Environmental Effects Monitoring. The 2005 technical guidance document (TGD) remains applicable to Cycle 5 (Environment Canada 2005), although updated guidance based on the 2008 PPER amendments is in progress.
Crofton EEM Cycle Five 1-1 Hatfield This report presents results from the EEM Cycle Five program for Catalyst Paper, Crofton Division. The program, previously described in the study design (Hatfield 2009), included a fish survey (using bivalves), benthic community survey, supporting sediment and water quality surveys, fish tissue study (crabs and sediments), and sublethal toxicity testing of mill effluent. Information on changes to mill processes, effluent treatment, and/or the receiving environment that occurred during Cycle Five also is presented. Sections in this report include:
Section 2 – Mill, Study Area, and Cycle Five Design Update;
Section 3 – Sublethal Toxicity Testing of Mill Effluent;
Section 4 – Fish Population Survey;
Section 5 – Fish Tissue Survey;
Section 6 – Benthic Invertebrate Community Survey;
Section 7 – Conclusions;
Section 8 – Closure;
Section 9 – References;
Section 10 – Glossary; and
Appendices.
Crofton EEM Cycle Five 1-2 Hatfield 2.0 MILL, STUDY AREA, AND CYCLE FIVE DESIGN UPDATE
2.1 MILL OPERATIONS
2.1.1 Process Description and Update
Catalyst Paper, Crofton Division operates a pulp and paper mill adjacent to Stuart Channel near the town of Crofton, British Columbia (Figure 2.1). Crofton is situated on the southeast coast of Vancouver Island north of Maple Bay, south of Chemainus, and across from Saltspring Island. The mill has been in operation since 1957, and currently produces Kraft pulp, newsprint, and directory paper.
The mill uses a newsprint furnish consisting of 66% mechanical pulp (TMP), 26% de-inked pulp, and 8% fillers (starch and precipitated calcium carbonate). The mechanical pulp wood furnish consists of 97% hemlock and 3% spruce/pine/balsam fir. The kraft pulp wood furnish consists of 35% western red cedar, 23% Douglas fir, 20% spruce/pine/balsam fir, 17% hemlock, and 5% cypress.
Average production in 2009 was 730 ADMt/d, and effluent flow was 92,700 m3/day (Table 2.1, Figure 2.2).
Detailed descriptions of mill processes, including bleaching, are documented in the Cycle One pre-design report (Hatfield 1994). Some of the key process updates that have occurred since the mill began operating in 1957 include:
Twinning of the outfalls in 1959;
Introduction of an effluent primary clarifier in 1978;
Introduction of a 24-hr activated sludge effluent system in 1982;
Introduction of chlorine-dioxide bleaching substitution in 1985;
Implementation of a modernization program to improve effluent quality between 1988 and 1998;
Installation of a UNOX secondary effluent treatment system (activated sludge breakdown using microorganisms) in 1992; and
Complete elimination of elemental chlorine bleaching at the mill in 1996.
Operational updates that occurred at the mill during Cycle Five included:
Shutdown of Paper Machine #1 from February 1 to May 25, 2009; and
Complete shutdown of the kraft mill from March 11 to September 30, 2009. Since restarting in October 1, 2009, the kraft mill has operated in a single-line configuration (half its two-line capacity).
Crofton EEM Cycle Five 2-1 Hatfield Figure 2.1 Location of Catalyst Paper Corporation, Crofton Division, on Stuart Channel, British Columbia.
123°40'W 123°35'W
LEGEND
T ri nc Lakes / Ponds om al Galiano i C ha Island Rivers / Streams Kuper n ne Island l
Pulpmill
H
o
u
s t
o Depth (metres)
n
P Intertidal
a s s 0 - 20 20 - 50 50 - 100 S tu 100 - 150 a rt C h Saltspring 48°55'N 150 - 200 a n n Island
48°55'N e l 200 - 250
W i lly
I. C h e m a Diffuser in u s R .
Osborn Crofton Bay Booth Catalyst Paper Bay Corporation Crofton Division Vancouver 0120.5 Island Km Scale 1:100,000 Projection: Albers Equal Area - NAD83 t
K:\Data\Project\CR1327\GIS\_MXD\CR1327_01_Location_20081009.mxd 123°45'W 123°40'W 123°35'W Table 2.1 Annual results for process effluent quality variables, Catalyst Paper, Crofton Division, 2007 to 2009.
Parameter 2007 2008 2009 Production Total Production (ADt/day) 2,206 2,103 730 Treated Effluent Quality Effluent Flow (m3/d) 144,900 142,300 92,700 pH 6.2 6.7 6.6 TSS (kg/day) 2,731 3,095 1373
BOD5 day (kg/day) 864 1,012 530 AOX (kg/day) 448 408 322 AOX (kg/ADt) 0.34 0.32 0.46 Toxicity (LC50)1: Daphnia Magna (48h LC50) 100% 100% 100% Rainbow Trout (96h LC50) 100% 100% 100%
2,3,7,8-T4CDD (ppq) ND ND ND
2,3,7,8-T4CDF (ppq) ND ND ND ND = not detected. ADt = air dried metric tones. ppq = parts per quadrillion (pg/L). 1 Percentage of tests conducted where LC50 (effluent concentration that kills 50% of organisms) was >100%.
Crofton EEM Cycle Five 2-3 Hatfield Figure 2.2 Annual production and effluent flows from 1970 to 2009 Catalyst Paper Corporation, Crofton Division.
Kraft mill shutdown for 6.5 mos; PM 1 shutdown for 4 mos 2,500 180,000
2,250 Daily Production Effluent Flow 150,000 2,000
1,750 120,000 /d) 3 1,500
1,250 90,000
1,000
Production (ADt/d) 60,000 750 Effluent Flow (m
500 30,000 250
0 0 1970 1971 1972 1973 1974 1975 1976 1977 1978 1979 1980 1981 1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009
2.1.2 Effluent Quality
Catalyst Paper, Crofton Division, discharges treated final effluent from Kraft and newsprint processes into Stuart Channel from its dual-diffuser main outfalls. An additional smaller outfall collects clean cooling water from a hot-water bypass system and discharges it, without treatment, to the main outfalls.
Effluent quality variables are routinely measured following provincial and federal requirements; annual results for the Crofton mill are presented in Table 2.1 for Cycle Five (i.e., 2007 to 2009) and historical results are presented in Figure 2.2 and Figure 2.3. Mean annual pulp production levels at the mill generally increased between 1997 and 2007, while effluent discharge rates generally declined somewhat (Figure 2.3). In 2008, production declined, and in 2009 production was cut significantly due to a complete shutdown of the kraft mill in March for 6.5 months and a return to only half capacity for the remaining three months of the year; effluent discharge rates declined correspondingly.
Total suspended solids (TSS), biological oxygen demand (BOD), and concentrations of adsorbable organic halides (AOX) in effluent declined following the 1998 modernization program designed to improve effluent quality (Figure 2.3). TSS, BOD, and AOX continued to decline after the 1992 implementation of secondary effluent treatment, and the 1996 elimination of elemental chlorine bleaching at the mill. Between 1999 and 2008, levels of BOD and TSS remained relatively consistent, while concentrations of AOX continued to slowly decline. In 2009, due to significant production cuts, levels of TSS
Crofton EEM Cycle Five 2-4 Hatfield (1,373 kg/day) and BOD (530 kg/day) were lower than in previous years; AOX (0.46 kg/ADt) was within the range of values reported within the last decade (Table 2.1, Figure 2.3).
Tetra-chlorinated dioxins (i.e., 2,3,7,8-T4CDD) and furans (i.e., 2,3,7,8-T4CDF) were not detected in Crofton effluent at any time during Cycle Five (Table 2.1).
In order to remain in compliance with the PPER, mills are required to demonstrate no acute toxicity of effluent to rainbow trout (i.e., all LC50s – effluent concentrations that kill 50% of trout – must be greater than 100% v/v effluent). In Cycle Five, there was no acute toxicity of effluent to either rainbow trout or Daphnia magna; 100% of all tests passed (Table 2.1).
2.1.3 Spills to the Receiving Environment
The following spills to the receiving environment were reported by Catalyst Paper, Crofton Division between 2007 and 2009 (Cycle Five):
100 L of precipitated calcium carbonate slurry spilled into the ocean during a barge offload on February 26, 2009; and
In late September 2009, a new cedar chip pile was stored at the foreshore near #3 dock at the mill. On November 26, 2009, a sample of the chip pile leachate discharging to the ocean failed a trout toxicity test. Immediate action was taken to eliminate the discharge.
2.1.4 Study Area Updates
During Cycle Five there were no other major anthropogenic influences or significant natural ecological variations in the Crofton EEM study area.
2.2 CYCLE FIVE STUDY DESIGN UPDATE
No major changes were made to the Cycle Five study design during field surveys.
Crofton EEM Cycle Five 2-5 Hatfield Figure 2.3 Mean daily total suspended solids (TSS), biochemical oxygen demand (BOD), and adsorbable organic halides (AOX) in effluent, Catalyst Paper Corporation, Crofton Division, 1981 to 2009.
1988-1992: Program implemented 30,000 to improve effluent quality Kraft mill shutdown for 6.5 mos; PM 1 25,000 shutdown for 4 mos 1992: Implementation of 20,000 secondary effluent treatment
15,000 1996: Switch to 100% elemental
TSS (kg/d) TSS 10,000 chlorine-free bleaching
5,000 1981 1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 1988-1992: Program implemented to improve effluent quality 60,000
50,000
40,000 1992: Implementation of secondary effluent treatment 30,000
BOD (kg/d) BOD 20,000 1996: Switch to 100% elemental chlorine-free bleaching 10,000 1981 1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009
1988-1992: Program implemented 6.00 to improve effluent quality
5.00
) 4.00 1992: Implementation of secondary effluent treatment 3.00
2.00
AOX (kg/ADt AOX 1996: Switch to 100% elemental chlorine-free bleaching 1.00 no data 0.00 1981 1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009
Crofton EEM Cycle Five 2-6 Hatfield 3.0 SUBLETHAL TOXICITY TESTING OF MILL EFFLUENT
Summary of Sublethal Toxicity Testing (winter 2007 through summer 2009) for Port Crofton EEM Cycle Five: No effect of effluent on survival or growth of topsmelt (Atherinops affinis) larvae was observed; Effects on echinoderm fertilization were observed at a mean effluent concentration of 23.0% (IC25); Algal reproduction was affected at a mean effluent concentration of 1.55% (IC25); and Based on a 1% effluent concentration zone of 600 m from the Crofton outfall, maximum potential zones of sublethal effect from the effluent discharge point are <6 m for survival of fish, 26 m for invertebrate fertilization, and 388 m for algal reproduction.
Federal and provincial government regulations require pulp and paper mills to undertake toxicity testing as part of their EEM programs to determine potential lethality or inhibitory effects of their effluent on fish and fish habitat. Sublethal and acute lethality tests may indicate whether fisheries resources are being protected or impacted in areas adjacent to their effluent discharges. Current EEM regulations require the use of sublethal toxicity tests to help meet the following objectives (Environment Canada 2005):
Contribute to the field program as part of a weight-of-evidence approach;
Compare process effluent quality between mill types, and measure changes in effluent quality as a result of effluent treatment and process changes; and
Contribute to the understanding of a mill’s relative contribution to downstream water quality in multiple discharge situations.
Sublethal toxicity testing for Crofton EEM Cycle Five included the following tests, as stipulated in the EEM TGD for pulpmills which discharge to a marine receiving environment (Environment Canada 2005):
Fish early life stage development growth and survival tests using topsmelt (Atherinops affinis). This test was excluded from the EEM testing requirements in August 2008 as stated in amendments to the PPER (Government of Canada 2008);
Invertebrate fertilization toxicity test using an echinoderm (either the sand dollar Dendraster excentricus or the purple sea urchin Strongylocentrotus purpuratus); and
Algal reproduction toxicity test using a red marine alga (Champia parvula).
Fish and invertebrate sublethal toxicity testing for Crofton was undertaken by Cantest Ltd. (formerly Vizon SciTech, Vancouver, British Columbia). Champia tests were conducted by the Saskatchewan Research Council, Saskatoon SK. A summary of reported endpoints are included with this Cycle Five interpretive report (Appendix A1).
Crofton EEM Cycle Five 3-1 Hatfield 3.1 SUBLETHAL TOXICITY TEST METHODS
3.1.1 General Methods and Definitions
During Cycle One, quarterly tests were required for the year field studies were completed. Since Cycle Two, sublethal toxicity testing of process effluent must be conducted twice each year. Testing for Cycle Five was initiated in winter 2007 and continued until summer 2009.
In Cycle Five, test seasons assigned were not necessarily representative of the date the test was conducted. The first test period of each year (the “winter” test period) was carried out between March and May. The second test period of each year (the “summer” test period) was carried out between September and January. The apparent discrepancy in the naming of test seasons was due to delays that occurred in Cycle Four, as a result of scheduled retests and restrictions associated with test organism availability. Figures presented in this section provide both the test season name and actual test date to prevent any confusion. The naming discrepancy has not been corrected because it has no effect on the validity of the toxicity results, and because correcting the naming would require that two sequential test periods be conducted too close to each other. The primary intent of having two test periods per year is to ensure tests are evenly spaced within the cycle.
On each test date, a grab sample of effluent was collected by mill personnel, following methods described in the EEM TGD (Environment Canada 2005) and shipped to Cantest Ltd. Sublethal toxicity testing involved exposure of organisms to a series of effluent dilutions. All sublethal toxicity tests were conducted with controls to assess the background response of test organisms and determine the acceptability of the test using predefined criteria. In addition, in-house cultures were tested with a reference toxicant to monitor the health and sensitivity of the culture.
Sublethal toxicity tests report LC50 or IC25 endpoints. Fish larvae growth, algal reproduction and invertebrate fertilization tests provide an IC25 endpoint, which is an estimate of the concentration of effluent that causes 25% inhibition of a quantitative biological function, such as reproduction or growth. The fish larvae test also yields an LC50 endpoint, which is the effluent concentration that is lethal to 50% or more of the test organisms. Confidence limits are given for each value where possible.
A zone of effluent mixing was determined by a plume delineation study during the Cycle One pre-design study (Hatfield 1994). This survey determined the maximum extent of effluent concentrations of 1% (i.e., 100:1 dilution) or greater, potentially present in the receiving water environment. This 1% effluent zone originally was used to define the extent of near-field and reference areas to aid in selecting sites to conduct required environmental sampling.
The 1% effluent zone represents conditions of minimum dilution, maximum extent, and long-term average conditions (i.e., long-term effect of effluent discharge) (Environment Canada 2005), and therefore represents worst-case
Crofton EEM Cycle Five 3-2 Hatfield effluent dilution conditions. For Catalyst Paper, Crofton Division, the maximum extent of 1% effluent was defined as a radial distance of approximately 0.6 km from the pulpmill diffusers (Hatfield 1994).
A maximum potential zone of sublethal effect was calculated for each test species from the geometric mean of the IC25 or LC50 results, and the extent of the 1% effluent concentration zone, as per Environment Canada (2005). This potential zone of sublethal effect describes the area where the effluent concentration exceeds the geometric mean of the IC25 or LC50 result, and is the maximum distance from the effluent discharge where a specified effect may be expressed for a test species. This maximum potential zone of sublethal effect was calculated as follows:
Extent of 1% effluent zone (m) Zone (m) = Geometric mean of IC25 or LC50 results
3.1.2 Sublethal Toxicity Test Methods
General procedures for conducting the topsmelt tests were based on Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to West Coast Marine and Estuarine Organisms, First Edition, EPA/600/R95/136, August 1995 (US EPA 1995). This 7-day static renewal test uses 9- to 15- day-old tospmelt (Atherinops affinis) larvae to assess the toxicity of a sample by comparing the growth and survival of exposed organisms to that observed in control organisms. The endpoints are effluent concentrations that result in 50% survival for seven days (LC50), and 25% inhibition of growth measured as dry weight (IC25) relative to controls.
General procedures for the echinoderm fertilization test are based on the methodology document Biological Test Method: Fertilization Assay Using Echinoids (Sea Urchins and Sand Dollars), Report EPS 1/RM/27, December 1992, and November 1997 amendments (Environment Canada 1997b). The test assesses the fertilization success of an echinoderm using either sand dollar (Dendraster excentricus) or sea urchin (Strongylocentrotus purpuratus). Male and female gametes are exposed to the test material for 20 minutes; percent fertilization is compared between the controls and the sample concentrations to determine if any significant inhibition of fertilization is observed. The IC25 endpoint is the percent effluent concentration where fertilization is reduced by 25% from control fertilization rates.
Procedures for conducting the marine algae (Champia parvula) reproduction test are based on Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Water to Marine and Estuarine Organisms, Third Edition, EPA 821/R/02-014, October 2002 (US EPA 2002). The algal reproduction test is a static, non-renewal, test where male and female plants are exposed to a test sample for a 48-hour period, followed by a 6- to 8-day recovery period. The effluent concentration endpoint (IC25) used to assess toxicity is the inhibition of cystocarp reproduction by 25% by the end of the recovery period compared to the control.
Crofton EEM Cycle Five 3-3 Hatfield 3.2 RESULTS AND DISCUSSION
Catalyst Paper, Crofton Division pulpmill conducted six sublethal toxicity tests from winter 2007 through summer 2009 for Cycle Five. The following issues were associated with Cycle Five testing:
Winter 2007 and summer 2007 invertebrate tests were repeated due to poor culture health in the original tests; and
Summer 2009 invertebrate test. A higher than usual sperm: egg ratio was required in the control to achieve an egg fertilization rate that met test protocol criteria. Despite the higher ratio, egg fertilization rates observed during testing were much lower than control rates, though still within test protocol criteria.
3.2.1 Topsmelt Growth and Survival Test
Topsmelt growth and survival tests were completed for the winter 2007 through summer 2008 test periods. Consistent with amendments made to the Pulp and Paper Effluent Regulations (Government of Canada 2008), this test was no longer required after winter 2008. For Crofton pulpmill effluent, the topsmelt LC50 (survival) and IC25 (growth) results were >100% (i.e., the highest concentration of effluent tested resulted in no observed effect), indicating no toxicity during the Cycle Five testing period (Figure 3.1, Figure 3.2). These results are similar to those observed in Cycle Four (Hatfield 2007).
Figure 3.1 Effect of exposure to Catalyst Paper, Crofton Division effluent on topsmelt survival, expressed as LC50 ±95% confidence limits, EEM Cycle Five.
>100 >100 >100 100
80
60
40 LC50 (% effluent)
20 Test no longer required
0 13-Apr-07 4-Sep-07 31-Mar-08 12-Jan-09 7-Apr-09 24-Nov-09
Winter 2007 Summer 2007 Winter 2008 Summer 2009 Winter 2009 Summer 2009 Sublethal Toxicity Testing Period (with actual test date)
Crofton EEM Cycle Five 3-4 Hatfield Figure 3.2 Effect of exposure to Catalyst Paper, Crofton Division effluent on topsmelt growth, expressed as IC25 ±95% confidence limits, EEM Cycle Five.
>100 >100 >100 100
80
60
40 IC25 (% (% effluent) IC25
20 Test no longer required
0 13-Apr-07 4-Sep-07 31-Mar-08 12-Jan-09 7-Apr-09 24-Nov-09
Winter 2007 Summer 2007 Winter 2008 Summer 2009 Winter 2009 Summer 2009 Sublethal Toxicity Testing Period (with actual test date)
3.2.2 Echinoderm Fertilization Test
Echinoderm fertilization results for the summer 2006 test period (Cycle Four) are included in this report (Figure 3.3); these results were excluded from the Cycle Four report because brine controls failed to meet the minimum survival criteria and results were unavailable at the time the report was finalized. The echinoderm fertilization IC25 for the summer 2006 retest was 10.5% v/v effluent.
Echinoderm (sand dollar or sea urchin) fertilization IC25s ranged from 4.1% to 55.5% v/v effluent in Cycle Five, with a geometric mean of 23.0%. Results were variable across test periods with no apparent increasing or decreasing trends, and were similar to those observed in Cycle Four. As described above, a higher than usual sperm:egg ratio was required during testing in the summer 2009 test period to achieve an egg fertilization rate in the control that met test protocol criteria. Egg fertilization rates observed during testing were within test protocol requirements, but were much lower than observed in the controls. As a result, the summer 2009 test (4.1% v/v effluent) may falsely indicate an elevated level of toxicity. Dose-response curves were relatively consistent across all test periods (Appendix A1).
Crofton EEM Cycle Five 3-5 Hatfield Figure 3.3 Effect of exposure to Catalyst Paper, Crofton Division effluent on echinoderm fertilization, expressed as IC25 ±95% confidence limits, EEM Cycle Five.
100
80
60 55.5 47.1
40 30.1 IC25 (% IC25 (% effluent) 21.0 22.3 20 10.5 4.1 0 5-Mar-07 7-May-07 9-Oct-07 31-Mar-08 12-Jan-09 7-Apr-09 24-Nov-09
Summer 2006 Winter 2007 Summer 2007 Winter 2008 Summer 2008 Winter 2009 Summer 2009
Cycle Four Sublethal Toxicity Testing Period (with actual test date)
3.2.3 Algal Reproduction Test
IC25 results and confidence limits for Cycle Five tests for algal reproduction using Champia parvula are summarized in Figure 3.4. Reproduction IC25 values ranged from 0.80% to 2.56% v/v effluent for a geometric mean of 1.55%. Results from Cycle Five indicate an increased effect of effluent on algal reproduction compared with all other cycles. Slight enrichment responses (enhancement of growth caused by nutrients present in the effluent) were observed at low effluent concentrations (<0.8% v/v effluent) during winter 2007, summer 2008, and winter 2009, testing periods. IC25 endpoints were variable during Cycle Five, and no increasing or decreasing trends were apparent. Overall, dose-response curves were relatively consistent during Cycle Five (Appendix A1).
Crofton EEM Cycle Five 3-6 Hatfield Figure 3.4 Effect of exposure to Catalyst Paper, Crofton Division effluent on algal reproduction, expressed as IC25 ±95% confidence limits, EEM Cycle Five.
100
80
60
40 IC25 (% (% IC25 effluent)
20
2.43 1.44 1.60 1.20 0.80 2.56 0 16-Apr-07 4-Sep-07 31-Mar-08 12-Jan-09 25-May-09 24-Nov-09
Winter 2007 Summer 2007 Winter 2008 Summer 2008 Winter 2009 Summer 2009
Sublethal Toxicity Testing Period (with actual test date) 3.2.4 Potential Zone of Sublethal Effect
Table 3.1 presents the geometric means of IC25 and LC50 results for each test species for all cycles, and the resulting Potential Zone of Sublethal Effect calculated using the defined 1% effluent zone (600 m). Calculations of geometric means and Potential Zones of Sublethal Effects can be found in Appendix A1.
The potential zone of sublethal effect for topsmelt cannot be calculated with any accuracy, as the IC25 and LC50 concentrations were always greater than the highest concentration of effluent tested. Consequently, the zone is shown as being less than the distance calculated (<6.0 m) assuming that the IC25 or LC50 was equal to the highest concentration tested.
The zone of sublethal effect increased slightly for echinoderms in Cycle Five (26.0 m) compared to Cycle Four (25.8 m) signifying a slight increase in toxicity; however, overall results since Cycle One indicate a decrease in toxic effect on echinoderm fertilization (Figure 3.5). Since Cycle One, the zone of sublethal effect for echinoderm fertilization has decreased from 37.5m to 26.0 m (Table 3.1). The potential zone of sublethal for algal reproduction effect increased from 219 m in Cycle Four to 388 m in Cycle Five. This zone of sublethal effect is the largest for Cycle Five, and has increased since Cycle One, signifying an increase in toxicity over time (Table 3.1, Figure 3.5).
Crofton EEM Cycle Five 3-7 Hatfield Table 3.1 Potential Zone of Sublethal Effect, Catalyst Paper Corporation, Crofton Division, EEM Cycle Five.
IC25 or LC50 Geometric Mean (% v/v) Potential Zone of Sublethal Effect 1(m) Sublethal Toxicity Test Species Cycle Cycle Cycle Cycle Cycle Cycle Cycle Cycle Cycle Cycle 1 2 3 4 5 1 2 3 4 5 Atherinops affinis2 Growth 67.1% 67.3% 77.9% 99.6% >100% 9.0m 8.9m 7.7m 6.0m <6.0m Survival 66.8% 67.3% 77.9% >100% >100% 9.0m 8.9m 7.7m <6.0m <6.0m Echinoderm Fertilization 16.0% 36.7% 17.7% 23.2%3 23.0% 37.5m 16.3m 33.8m 25.8m3 26.0m Champia parvula 4.5% 44.5% 7.9% 2.7% 1.6% 135m 13.5m 76.2m 219m 388m Reproduction
1 Based on 1% effluent zone of 2,000m. 2 Cycle Five geomeans and Potential Zones of Sublethal Effect for topsmelt growth and survival are based on three test periods. Testing for this species was no longer required after winter 2008. 3 Echinoderm fertilization test results for the summer 2006 test period were excluded from this calculation in the Cycle Four Interpretive Report because retest results were unavailable at the time the report was finalized. Values for Cycle Four presented here have been updated to include these results.
3.3 CONCLUSIONS
Toxicity testing results contribute to the overall weight-of-evidence used to assess potential environmental effects of effluent discharges. When interpreting sublethal toxicity data it is important to keep in mind that laboratory toxicity test results may not accurately predict toxicity in receiving environments. Laboratory tests involve single species that may or may not be found in the study area, and there is an assumption that there is no background toxicity in the area. Furthermore, effluent plume dispersion and dilution vary due to tides and currents, and receiving waters and effluent quality characteristics vary seasonally or in other ways that could affect toxicity levels.
Figure 3.5 Geometric means of IC25 and LC50 results from sublethal toxicity tests of Catalyst Paper, Crofton Division effluent for EEM Cycle One through Cycle Five.
Cycle One Cycle Two Cycle Three Cycle Four Cycle Five 100
80
60
40
20 Percent Effluent Concentration Percent 0 Topsmelt early life stage Topsmelt early life stage Echinoderm Algal survival (LC50) growth (IC25) survival (IC25) reproduction(IC25)
Crofton EEM Cycle Five 3-8 Hatfield The quality of effluent discharged from the Crofton pulpmill, as measured by sublethal toxicity results, was similar to effluent quality observed in previous EEM cycles for topsmelt and echinoderm. However, effluent quality has decreased for algal reproduction relevant to previous cycles (Table 3.1, Figure 3.5). Effluent did not affect fish early life stage growth or survival, echinoderm fertilization was affected at a mean effluent concentration of 23.0%, and algal reproduction was affected at a mean effluent concentration of 1.55%.
The sublethal toxicity testing results indicate that Crofton effluent may influence the receiving environment in a zone up to a calculated distance of 388 m from the pulpmill diffuser.
Crofton EEM Cycle Five 3-9 Hatfield 4.0 FISH POPULATION SURVEY
Summary of Fish Population Survey for Crofton EEM Cycle Five: The overall objective of the fish population survey was to determine the effects of effluent exposure on fish populations near the Crofton mill. The study objective was achieved by sampling mussels and oysters at three stations: CRO3A (near-field), CRO5A (far-field), and CRO4 (far-far-field reference). The control/impact design was used to determine if significant differences (i.e., effects resulting from effluent exposure) could be detected; Lower densities of older, larger, heavier mussels were present closer to the mill, indicating greater survival, energy use, and energy storage in these organisms relative to those at the reference station; Reproductive energy use did not show any significant differences between near-field and reference mussels; In contrast to mussels, oysters in the near-field demonstrated lower survival, energy use, and energy storage than reference oysters. Oysters at the near-field station were significantly smaller and lighter, both in general and for their age, and were present in higher densities relative to other stations; Ratios of male versus female mussels were similar between the near-field and reference areas, and GSI did not demonstrate any significant difference between the two areas, indicating that sex did not play a role in influencing the significantly greater survival, energy use, and energy storage of mussels in the near-field; Mussels were older in the near-field, indicating that recent spawning and/or spat settlement was not as successful at the near-field station; In contrast to mussels, the youngest oysters in the study were only found at the near-field station, and in terms of their size-at-age the near-field oysters were substantially smaller; Coexistence of mussels and oysters can be difficult, with the larger oysters often colonizing suitable habitat at the expense of mussels; given the more favourable growing conditions observed for oysters at the reference station (i.e., higher temperature and DO), and the presence of larger oysters and smaller mussels at this station, it is possible that oyster populations have settled at the reference station at the expense of mussels; and In contrast to previous cycles, the response-based effects summary for Cycle Five demonstrated significant effects between the exposure and reference areas for both mussels and oysters. Given that opposing effects were observed for the two species, it is difficult to say whether the differences are associated with natural or mill-related factors.
4.1 INTRODUCTION
Mills are required to evaluate the effects of effluent on fish populations if the effluent is diluted to a concentration of more than 1% of release within 250 m of the discharge point (Environment Canada 2005). At Crofton, the 1% zone of effluent concentration has been defined to be an elliptical area approximately 600 m in diameter around each outfall on the main diffuser (Figure 4.1).
Health indicators were measured in wild Pacific oysters during the fish survey at Crofton for Cycles One, Two, and Three. During Cycles Two and Three, oysters did not exhibit effects associated with the exposure gradient in the vicinity of the Crofton mill (Hatfield 2000, 2004). No fish survey was required for Crofton EEM Cycle Four; as per technical guidance, given no effect was observed during two consecutive fish surveys (Cycles Two and Three) this component could be repeated in six years (Cycle Five).
Crofton EEM Cycle Five 4-1 Hatfield In Cycle Five, Pacific oysters were again used as a sentinel species for the EEM fish survey, in order to facilitate comparisons with historical results. The Cycle Five design was a repeat of Cycle Three, evaluating the same oyster parameters within the same three areas of the study zone (i.e., near-field, far-field, far-far-field). EEM guidance requires the use of two sentinel fish species in order to fulfill the requirements of the fish survey (Environment Canada 2005). Given these requirements, as well as recent scientific and technical improvements in the use of bivalve mollusks as environmental monitors, Pacific blue mussels (Mytilus trossolus) were used as the second sentinel species during the Crofton EEM Cycle Five fish survey.
4.2 METHODS
4.2.1 Modifications to Sampling Design
No other modifications were made to the Crofton EEM Cycle Five design (Hatfield 2009a).
4.2.2 Control/Impact Design
Oysters and mussels were collected at one station from each of the three key zones along a gradient of expected effluent exposure: one near-field station, one far-field station, and one far-far-field (reference) station (Table 4.2; Figure 4.1). The control/impact study used analysis of variance (ANOVA) and analysis of covariance (ANCOVA) to determine differences whole organism metrics and indices between stations. This approach allowed for within-station variability to be determined.
Table 4.1 Bivalve survey control/impact sampling design, Crofton EEM Cycle Five, 2009.
Distance from Intertidal Station Area Effluent Exposure Level Outfalls CRO4 (Dayman Island) 9.6 km Far-far-field Considered to be reference CRO5A (North Reef) 3.1 km Far-field Exposure outside 1% effluent zone CRO3A (Bisco Island) 0.4 km Near-field Exposure within 1% effluent zone (600 m)
Crofton EEM Cycle Five 4-2 Hatfield 4.2.3 Sampling Locations and Collection Dates
Oysters and mussels were collected at identical sampling locations during the Cycle Five fish survey to allow for comparison of endpoints among species; locations also corresponded with control/impact stations sampled in Cycle Three in order to allow comparisons over time. Mussels were sampled in spring (March) 2009; oysters were sampled in summer (July) 2009. Sampling locations and collection dates are presented in Table 4.2.
Table 4.2 Bivalve survey sampling locations and collections, Crofton EEM Cycle Five, 2009.
Subsamples Submitted Station Collection Date Coordinates Samples Collected for Analyses CRO4 11-Mar-2009 48° 58.341 40 blue mussels 40 individual tissues: dry weight (ALS) (Dayman Island) 123° 41.561 Habitat Description 1 composite x 40 tissues: lipids (ALS) Water Quality 40 individual shells: aging (UVic) 6-Jul-2009 48° 58.529 64 Pacific oysters 64 individual tissues: dry weight (ALS) 123° 41.623 Habitat Description 1 composite x 64 tissues: lipids (ALS) Water Quality 64 individual shells: aging (UVic)
CRO5A 12-Mar-2009 48° 53.175 40 blue mussels 40 individual tissues: dry weight (ALS) (North Reef) 123° 38.338 Habitat Description 1 composite x 40 tissues: lipids (ALS) Water Quality 40 individual shells: aging (UVic) 15-Jul-2009 48° 54.857 64 Pacific oysters 64 individual tissues: dry weight (ALS) 123° 37.621 Habitat Description 1 composite x 64 tissues: lipids (ALS) Water Quality 64 individual shells: aging (UVic) CRO3A 12-Mar-2009 48° 53.175 40 blue mussels 40 individual tissues: dry weight (ALS) (Bisco Island) 123° 38.338 Habitat Description 1 composite x 40 tissues: lipids (ALS) Water Quality 40 individual shells: aging (UVic) 15-Jul-2009 48° 52.771 65 Pacific oysters 65 individual tissues: dry weight (ALS) 123° 37.760 Habitat Description 1 composite x 65 tissues: lipids (ALS) Water Quality 65 individual shells: aging (UVic)
Crofton EEM Cycle Five 4-3 Hatfield Figure 4.1 Pacific oyster and mussel sampling locations, Crofton EEM Cycle Five, 2009.
123°40'W 123°35'W
LEGEND
Waterbody
49°0'N Stream Network
Pulpmill
T r in Depth (metres) co ") CR04 m a ") l Intertidal i C h a n 0 - 20 n e Kuper l 20 - 50 H 50 - 100 Island o
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s 100 - 150
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o
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s Sampling Location ") Pacific Oyster (March 2009) ") Mussel S (July 2009) tu a CR05 rt Saltspring C h ")")
48°55'N a Island n n e l
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I.
Diffusers
R . ") CR03 ")
Osborne Crofton Bay Booth Bay Catalyst Paper Corporation Crofton Division
Vancouver 0120.5 Island Km Scale 1:100,000 Projection: Albers Equal Area - NAD83 48°50'N t
K:\Data\Project\CR1327\GIS\_MXD\B_InterpretiveReport\ 123°40'W 123°35'W CR1327_B07_OysterMussel_20100112.mxd As in previous bivalve surveys, Cycle Five sampling of oyster and mussel communities was timed to be shortly in advance of their anticipated spawning periods in the Crofton area. For oysters, this spawning event can take place any time between late June and early September, but most typically occurs in late July or early August (Quayle 1988). Sampling was therefore performed in early July. Wild Pacific blue mussels (Mytilus trossolus) were collected in March 2009 during benthic invertebrate sampling, in advance of their expected spawning period which is known to occur in the Strait of Georgia during April or May (Cowles 2005).
4.2.4 Field Sampling Procedures 4.2.4.1 Oyster Collections
Pacific oysters were collected at low tide from the intertidal zone at bivalve collection stations on July 6 and 15, 2009. 65 oysters of all sizes were collected by hand from each of the three stations semi-randomly, using the following sampling methods:
A 0.25 m2 (0.5 x 0.5 m) quadrat was placed randomly (i.e., thrown behind the back) along the rocky, intertidal shore at each station;
All oysters (i.e., of all sizes) within the quadrat were collected, except those that were tightly cemented to substrate and therefore could not be effectively sampled without destroying the organism;
Quadrats were placed until a total of 65 oysters were collected from each station; and
The numbers of quadrats used was recorded to allow calculation of oyster density at each station.
All collected specimens were placed into buckets, and kept alive with cool seawater and a bubbler until they could be dissected (within 24 hours).
4.2.4.2 Mussel Collections
Although Mytilus trossolus were the target wild blue mussel species, it should be noted that the non-indigenous blue mussel species, Mytilus edulis, is also present in coastal British Columbia (Pomeroy and Goyette 1983). The two species (M. trossolus and M. edulis) are not easily distinguishable without DNA analyses (Salazar 2000), and therefore may be sampled in the field as one species. Sampling both species as one should not pose any problems when it comes to analyses based on non-reproductive measurement endpoints. However, differences may exist in the reproductive biology of each species, specifically with respect to the duration of their respective spawning periods (St-Jean et al. 2003). Sampling in advance of spawning periods for both species alleviated this as an issue, and allowed for meaningful comparisons of mussel reproductive condition between stations, regardless of species.
Crofton EEM Cycle Five 4-5 Hatfield Pacific blue mussels (M. trossolus and M. edulis) were collected at low tide from the intertidal zone at bivalve collection stations on March 11 and 12, 2009. Forty mussels of all sizes were collected by hand at each of the three sampling stations using the same semi-random sampling methods used to collect oysters (see Section 4.2.4.1). Mussels were kept cool and wet/moist until they could be dissected that same day.
4.2.4.3 Supporting Environmental Variables
Supporting environmental variables (i.e., water quality and habitat characteristics) were measured twice at each of the three bivalve collection stations: once during the March 2009 mussel survey, and once during July 2009 oyster survey. Continuously-recording thermographs were installed at each station in March 2009 and left in situ until oyster collections in July, in order to develop water temperature profiles.
Habitat Characteristics
Physical habitat characteristics (i.e., substrate type, macrophytic coverage, slope, and aspect) were measured within the intertidal bivalve collection habitat at each station during low tide. Slope was measured using a clinometer, and aspect was determined using a GPS compass. General features of each station were noted and photo-documented. Water Quality
Ambient sea water was sampled at each intertidal bivalve collection station for the following measures using a YSI water quality meter:
temperature (±0.1°C); salinity (±0.1 ppt); and dissolved oxygen (±0.01 mg/L).
The YSI probe was lowered 0.5 m into surface water near the shore at each station.
Thermographs
One continuously-recording thermograph (HOBO Water Temp Pro) was installed at each bivalve collection station in order to obtain a series of water temperature profiles between the spring (March) and summer (July) seasons. Thermographs were installed during low tide at a depth of approximately 0.5 m below the lowest low tide line, and were fixed in place using rope and weights.
4.2.5 Laboratory Procedures 4.2.5.1 Oyster Dissections
General oyster dissection methods followed those of previous recent studies (e.g., Hatfield 2000, 2004). Dissections were conducted in a climate-controlled facility to maximize precision of measurements. Dissection equipment was cleaned between each organism dissection.
Crofton EEM Cycle Five 4-6 Hatfield The surface of each oyster was scrubbed to remove foreign material. Each individual’s length, width, and depth (mm) were recorded using calipers; whole weight of the organism (± 0.1 g) also was measured. Volume (±5 mL) of the whole oyster was determined by water displacement in a measured beaker.
The oyster was then opened and all internal tissue and liquid removed to a pre-labelled, pre-weighed glass sample jar and weighed (±0.1 g). Tissues were then frozen prior to shipment to ALS Environmental Ltd. (Vancouver, BC) for determination of individual dry-weights and station-composite lipids, following methods outlined in Section 4.2.5.3. Volume of the empty valves (±5 mL) was then determined by displacement in the beaker, and weights were taken (±0.1 g). Valves for one organism were then placed in a Ziploc bag and labelled with an individual identification number.
Hermaphrodism and changes of sex in oysters is common (Quayle 1988), and the separation of the gonadal mass from the visceral mass is impractical and limits accurate estimation of gonad size for indirect estimation of fecundity. Despite attempts, it was not possible to sex oysters based on visual assessment (e.g., colour, size of gonadal tissues) given these tissues were non-distinct and intermingled with the digestive gland. Therefore, oyster sex and fecundity was not evaluated for the Cycle Five survey.
4.2.5.2 Mussel Dissections
Dissections were conducted in a climate-controlled facility to maximize precision of measurements. Dissection equipment was cleaned between each organism dissection.
Individual mussels were cleaned of external foreign material, then measured for shell length, width, and girth (±1 mm) using calipers; whole weight of the organism (±0.001 g) also was measured using an electronic balance. Mussels were then placed on a stainless-steel tray for dissection.
Each mussel was opened and the mantle (which includes gonadal tissues) was identified, removed to a pre-labelled, pre-weighed glass sample jar, and weighed (±0.001 g) using an electronic balance. Sex and maturity were determined based on gonad colour and size.
Remaining internal tissues and liquid were removed, added to the sampling jar containing the gonadal tissues, and weighed (±0.001 g). Tissues were then frozen prior to shipment to ALS Environmental Ltd. (Vancouver, BC) for determination of individual dry-weights and station-composite lipids, following methods outlined in Section 4.2.5.3. Empty valves for one mussel were weighed (±0.001 g), and then placed in a Ziploc bag labelled with an individual identification number. Valves were shipped to the University of Victoria for aging analysis, following methods outlined in Section 4.2.5.4.
Crofton EEM Cycle Five 4-7 Hatfield 4.2.5.3 Dried Tissue Weights and Lipids
ALS Environmental Ltd. (Vancouver, BC) oven-dried individual oyster and mussel tissue samples at 70°C. After a 48-h drying period, tissues were removed from the oven and allowed to cool for ten minutes. Dried meats for each individual were then weighed and the results reported. ALS also analyzed one composite sample of dried meat for each species from each station (i.e., 40 mussels and 65 oysters) for percent lipids.
4.2.5.4 Age Analysis
All aging analyses were undertaken by Ms. Dawna Brand of the University of Victoria (UVic), Biology Department.
Oysters and mussels were aged following procedures described in Anwar et al. (1990) and Richardson et al. (1970) along with the following modifications. Radial sections of Blue mussel and Pacific oyster shells were cut in a plane from the umbo to the growing point of the shell. Shells were then polished on the cut edge using successively finer grit wet dry paper. Polished shells were burned by lightly passing thru a flame, then coated with immersion oil and examined using a dissecting microscope. Age was determined by growth ring counts at the umbo, at the thickest end of the shell and along the cut plane from the umbo to the growing point. For oysters muscle scars were also used for aging. For QA/QC each shell was aged independently three times. An electronic spreadsheet (MS Excel format) of data including station location, shell ID and age was prepared. Given oyster and mussel shells exhibit a high degree of checking, flaking and powdering, only the rings on the chondrophore are reliable for determining growth. Due to the number of check marks present, growth is determined more easily with a dissecting scope, with rings counted at a lower magnification, similar to the break and burn method employed for fish otoliths. Ages were recorded up to the year of growth the oysters or mussels were in at time of collection.
4.2.6 Analytical Approach 4.2.6.1 Data Handling
Excel was used to format data, calculate bivalve metrics and indices, and generate descriptive statistics and graphs. All statistical analyses were performed using SYSTAT v.10 statistical software (SPSS Inc. 2000), and power analyses were conducted using G*Power software (Faul and Erdfelder 1992). Separate analyses were conducted for each species (mussels and oysters) and sex (mussels only).
4.2.6.2 Community Metrics
Excel was used to determine densities of mussels and oysters (number/m2) at each station using counts of mussels and oysters collected per quadrat, and the total number of quadrats collected per station.
Crofton EEM Cycle Five 4-8 Hatfield 4.2.6.3 Whole-organism Metrics
A number of whole-organism metrics were tabulated in Excel and used in data analyses for both mussels and oysters, including:
Length, width, and girth (mm);
Whole wet (total body) weight (g);
Dry meat weight (g);
Shell weight (g);
Whole volume (ml);
Cavity volume (ml);
Shell volume (ml); and
Age (yrs).
Internal cavity volume for oysters was calculated as whole volume less shell volume, which were directly measured through volume displacement during dissections (see section 4.2.5.1). Cavity volume for mussels (mm3) was estimated as "shell length x shell width x shell depth" divided by a correction factor of 5.0; the mm3 volume was multiplied by 0.001 to obtain mL units. This method of estimating cavity volume has been used successfully for mussels collected at other BC coastal mills. Regression analyses indicate that estimated cavity volume correlates significantly with actual cavity volume in blue mussels (r=0.971, r2=0.943, df=1, 123, p<0.0005). However, the estimated volume is approximately five times larger than the actual volume; hence a 5.0 correction factor is used.
4.2.6.4 Whole-organism Indices
A variety of whole-organism indices will be calculated in Excel and used to assess condition, growth, and survival in both mussels and oysters, including condition index, shell density, size-at-age, and gonadosomatic index (mussels only).
Condition Index
Condition Index (K) describes the relationship between a bivalve’s weight and the internal cavity volume, and ultimately is a measure of an organism’s fatness within its shell. Bivalve condition has been shown historically to be negatively affected by industrial and other types of marine pollution (Quayle 1988).
Condition was calculated as:
Dry weight of meat (g) x 1000 Condition (g/cm3) = Internal cavity volume (mL)
Crofton EEM Cycle Five 4-9 Hatfield Shell Density
Shell density describes the relationship between the valve weight and volume, and has been shown by some researchers to be affected by both natural and anthropogenic stresses (Brown and Hardwick 1988). His and Robert (1987) observed a thickening of oysters' shells in response to the introduction of antifoulant paints to harbours in the southeastern United States; Seaman (1985, cited in Brown and Hardwick 1988) observed holes and thinning in shells of oysters growing in areas of low salinity.
Given the unpredictable shape of oyster valves, shell density was calculated using direct measurements of valve weight and volume, as follows:
Dry valve weight (g) Oyster Shell Density (g/cm3) = Dry valve volume (cm3)
Given the predictable shape of mussel valves, and the difficulty in accurately their shell volume directly, estimated internal cavity volume was used as a proxy for shell volume in the following equation adapted from Aldrich and Crowley (1986):
Dry valve weight (g) x 1000 Mussel Shell Density (mg/cm3) = Internal cavity volume (cm3)
Size-at-age
Size-at-age describes the relationship between a bivalve’s weight and age, and was calculated as:
Dry weight of meat (g) Size– at-Age (mg/yr) = Age (yr)
Gonadosomatic Index
Gonadosomatic index (GSI), calculated by expressing gonad weight as a percentage of total meat weight (% gonadal tissue), was used to evaluate reproduction in mussels.
GSI was calculated as:
Mantle weight (g) x 100 GSI = Wet tissue weight (g)
GSI could not be determined for oysters, given their gonadal tissues are substantially intermingled with other tissues, making separation of the gonadal mass from the visceral mass impractical.
Crofton EEM Cycle Five 4-10 Hatfield 4.2.6.5 Statistical Analyses Summary Statistics
Summary statistics, including mean, median, standard deviations, standard error, and minimum and maximum values were calculated in Excel for each of the whole-organism metrics and indices.
ANOVAs
SYSTAT was used to perform two-tailed Analysis of variance (ANOVA) and Tukey’s multiple comparisons in order to compare oyster and mussel metrics and indices between stations CRO3A, CRO5A, and CRO4 to determine if there were differences between the near-field, far-field, and far-far-field areas, respectively. The following whole organism metrics and biological indices were used:
Age;
Dry meat weight;
Total body weight;
Shell weight; and
Shell density.
Analyses were conducted using data for individual bivalves. ANOVAs were performed using a significance level of α = 0.10, as required by the most recent technical guidance document (Environment Canada 2005). Residuals from the ANOVA were saved and qualitatively evaluated for normality and homogeneity of variance (i.e., ANOVA assumptions) using residual plots.
If data failed to meet ANOVA assumptions, analyses were conducted using log10-transformed variables. If assumptions of the model were not met using transformed variables, ANOVAs and Tukey’s comparisons were conducted using ranked data.
If large outliers were detected in the data set, they were removed and data was reanalyzed using the methods described above. Results were compared to non parametric analyses of the whole data set, to determine the effect of the outlier on the overall results.
Crofton EEM Cycle Five 4-11 Hatfield ANCOVAs
SYSTAT was used to generate analyses of covariance (ANCOVA) to test for differences in relationships between oyster and mussel indices (according to different covariates) between stations. Indices and their potential covariates that were tested included:
Size-at-age:
o dry meat weight (dependent variable) by age (covariate); and
o total body weight (dependent variable) by age (covariate).
Condition Index:
o dry meat weight (dependent variable) by total body weight (covariate); and
o dry meat weight (dependent variable) by cavity volume (covariate).
GSI (relative gonad weight):
o Gonad weight (dependent variable) by wet meat weight (covariate).
An assumption of the ANCOVA model is that the slopes of the regression lines are equal between areas; therefore, differences in slopes were tested prior to conducting the ANCOVA. Generally, ANCOVA is fairly robust even when slopes are not equal, so slopes were considered different when p<0.01 (Paine 1998). Data were log10-transformed where appropriate. As recommended in the updated EEM guidance (Environment Canada 2005), tests of significance used a probability level (α, two-tailed) of 0.10.
Response-based Effects Assessment
Biological variables were organized into broad subgroups, according to EEM technical guidance (Environment Canada 2005) and as adapted from a theoretical effects-assessment framework for finfish developed by Gibbons and Munkittrick (1994). Although this framework was not developed for application to shellfish, the same general principles apply. This framework can be used to determine types of effects that a stressor such as water pollution may have on bivalve populations, namely:
Survival – Response to a stressor that caused direct or indirect mortality of bivalves or reduced reproduction in the population would be manifested as an increase in mean age in an exposed population relative to reference bivalves;
Crofton EEM Cycle Five 4-12 Hatfield Energy use or expenditure – Response to a stressor that affected energy expenditure, either positively or negatively (e.g., through an increase or decrease in available food resources), would be manifested as differences in total weight or dry meat weight between exposed and reference populations, with these variables increasing or decreasing with increases or decreases in energy expenditure of bivalves examined; and
Energy storage – Response to a stressor that affected energy storage would be manifested as differences in condition between exposed and reference populations, with these variables indicating more or less storage of energy.
Table 4.3 summarizes how ANOVAs and ANCOVAs were used to analyze bivalve metrics and indices in order to evaluate effects associated with the Crofton mill.
Table 4.3 Statistical analyses performed to evaluate effects on bivalves, Crofton EEM Cycle Five, 2009.
Type of Dependent Statistical Parameter Covariate (X) Response Variable (Y) Procedure Survival Age Age None ANOVA Energy Size Dry Meat Weight None ANOVA Expenditure Total Weight None ANOVA Gonad size Gonad Weight Wet Meat Weight ANCOVA (mussels only) Size-at-age Dry Meat Weight Age ANCOVA Total Weight Age ANCOVA Energy Condition Dry Meat Weight Total Weight ANCOVA Storage Dry Meat Weight Cavity Volume ANCOVA Shell Condition Shell Weight None ANOVA Shell Density None ANOVA
Power Analyses
Power analysis was performed using G*Power in order to evaluate the possibility of false negative results (i.e., concluding that no difference in response exists when one actually does). Peterman (1990) has argued that the consequences of false negatives may be greater than the dangers of false positives (i.e., concluding a difference when none exists), and that environmental impact assessments should have adequate statistical power to detect meaningful differences. In other words, it is necessary to determine whether a comparison that did not show a statistical difference actually had sufficient power to detect a given difference, if one existed.
Crofton EEM Cycle Five 4-13 Hatfield Statistical power is a function of sample size, variability and magnitude of difference (i.e., effect size) one wishes to detect. The effect size is not easily defined. The Fish Survey Expert Working Group has recommended an effect size of ±25% in relative gonad size for EEM fish surveys (Environment Canada 2005). This approach was modified for use in the Crofton EEM Cycle Three program, and was used again in this Cycle Five study. Power analyses were conducted to evaluate whether there was sufficient power to detect a 25% difference in condition between populations for both oysters and mussels. The mean squared error (MSE) term from the ANCOVA statistical model provided the estimate of between-area variance in condition. Statistical comparisons were considered to have sufficient power (P=1-β, probability of detecting an effect size) when P≥0.90 (Environment Canada 2005). 4.3 RESULTS
4.3.1 Mussels 4.3.1.1 Community Metrics
In March 2009, 40 mussels were collected and analyzed from each of the three bivalve collection stations (i.e., CRO3A, CRO4, and CRO5A). Densities of mussels along shorelines at these stations were variable among stations. At far-far-field station CRO4 (Dayman Island), densities were very high (284 mussels/m2), and the target number of organisms was obtained using one quadrat sample. At stations CRO3A (Bisco Island) and CRO5A (North Reef), densities were so low (often zero to just one or two mussels) that randomly placed quadrats were not used to sample these stations and densities could not be calculated. 4.3.1.2 Whole-organism Metrics
Whole-organism metrics are presented in tabular form in Appendix A3 for each blue mussel collected at each station. Metrics reported include length, width, girth, whole wet weight, wet meat weight, wet mantle weight, dry meat weight, shell weight, cavity volume, sex, and age. 4.3.1.3 Whole-organism Indices
Whole-organism indices are presented in tabular form in Appendix A3 for each blue mussel collected at each station. Indices reported include condition index, shell density, size-at-age, and GSI. 4.3.1.4 Summary Statistics
Descriptive statistics summarizing characteristics measured in blue mussels from each sampling station are presented in Table 4.4 and Table 4.5. Size and Weight Distributions
Overall, mussels were largest and heaviest at station CRO5A (far-field), and smallest and lightest at station CRO4 (far-far-field) (Figure 4.2 and Figure 4.3). Weights throughout the study area ranged between 0.64 g and 7.94 g (Table 4.4); weight frequency distributions were slightly variable from station to station (Figure 4.4):
Crofton EEM Cycle Five 4-14 Hatfield Mussels from CRO3A exhibited a relatively normal, but slightly bimodal, weight frequency distribution curve; the highest frequencies of mussels fell within the 2-3 g and, to a lesser extent, the 4-5 g categories; Mussels from CRO5A exhibited a relatively normal weight frequency distribution curve, and the majority of mussels fell within the 3-4 g category; and Mussels from CRO4 exhibited a slightly positively skewed weight frequency distribution curve; frequencies of smaller mussels (< 2 g) were higher than mid- to larger-sized mussels.
Figure 4.2 Mean length, width and girth (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
Length (mm) Mean length (all stations) 35.0 Width (mm) Mean w idth (all stations) Girth (mm) Mean girth (all stations) 30.0
25.0
20.0
15.0 Size (mm) Size
10.0
5.0
0.0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Figure 4.3 Mean whole wet weight (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
4 Mean w eight (all stations) 3
3
2
2
1 Mean wet weight (g) weight wet Mean 1
0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Crofton EEM Cycle Five 4-15 Hatfield Table 4.4 Whole-organism metric summary statistics for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
Length Width Gir th Age Whole wet Wet Mantle Wt. Shell Wt. Dry Tissue Est Cavity Station Summary Statistic (mm) (mm) (mm) (yrs) weight (g) Tissue Wt. (g) (g) Wt. (g) Vol (ml) CRO3ASample size (n)40404040394034404040 Bisco Isl. Minimum 24.00 13.00 8.00 2.00 1.32 0.32 0.02 0.67 0.04 0.62 (near-field) Maximum 39.00 20.00 17.00 3.00 5.64 1.77 0.30 3.06 0.30 2.65 Median 29.00 16.00 12.00 3.00 2.75 0.80 0.10 1.24 0.13 1.08 Mean 28.73 16.00 12.40 2.95 2.99 0.84 0.12 1.38 0.13 1.18 Standard Dev 3.45 1.71 1.92 0.04 1.13 0.33 0.08 0.55 0.06 0.44 Standard Error 0.55 0.27 0.30 0.22 0.18 0.05 0.01 0.09 0.01 0.07 CRO5ASample size (n)40404039404040404040 North Reef Minimum 24.00 13.00 9.00 2.00 1.23 0.52 0.07 0.82 0.12 0.56 (far-field) Maximum 38.00 20.00 15.00 3.00 5.41 2.17 0.61 2.25 0.43 2.13 Median 30.00 16.00 12.50 3.00 3.06 1.20 0.23 1.43 0.26 1.21 Mean 29.88 16.43 12.45 2.90 3.11 1.25 0.25 1.48 0.26 1.25 Standard Dev 2.83 1.66 1.34 0.05 0.87 0.35 0.12 0.31 0.07 0.33 Standard Error 0.45 0.26 0.21 0.31 0.14 0.06 0.02 0.05 0.01 0.05 CRO4Sample size (n)40404040404036384040 Dayman Isl. Minimum 17.00 10.00 7.00 1.00 0.64 0.08 0.01 0.28 0.01 0.24 (far-far-field) Maximum 43.00 21.00 17.00 4.00 7.94 1.98 0.43 3.52 0.33 2.71 Median 24.50 13.00 10.00 2.00 1.63 0.42 0.03 0.75 0.06 0.65 Mean 24.98 13.23 10.20 2.23 1.98 0.50 0.06 0.90 0.08 0.75 Standard Dev 5.40 2.39 2.20 0.11 1.40 0.33 0.08 0.63 0.05 0.47 Standard Error 0.85 0.38 0.35 0.70 0.22 0.05 0.01 0.10 0.01 0.07
Crofton EEM Cycle Five 4-16 Hatfield Table 4.5 Whole-organism indices summary statistics for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
Summary Condition Index Shell Density Size-at-Age Station GSI Statistic (mg/cm3) (mg/cm3) (mg/yr)
CRO3A Sample size (n) 40 40 40 34 Bisco Island Minimum 36.01 885.58 11.67 3.70 (near-field) Maximum 179.69 1953.13 99.67 26.96 Median 118.53 1150.23 44.17 11.10 Mean 113.58 1172.96 44.88 12.69 Standard Error 5.03 30.38 3.01 0.89 Standard Dev 31.81 192.13 19.02 5.21 CRO5A Sample size (n) 40 40 39 40 North Reef Minimum 156.96 899.05 44.00 9.92 (far-field) Maximum 272.99 2306.55 143.33 33.13 Median 204.89 1179.88 86.67 18.05 Mean 208.16 1221.59 88.83 19.01 Standard Error 4.35 39.09 3.61 0.87 Standard Dev 27.52 247.25 22.55 5.52 CRO4Sample size (n)40384040 Dayman Island Minimum 28.65 599.11 11.00 0.00 (far-far-field) Maximum 158.85 1924.49 81.25 21.80 Median 103.67 1115.09 28.50 8.05 Mean 103.40 1153.57 32.79 8.06 Standard Error 3.98 35.35 2.31 0.78 Standard Dev 25.18 217.92 14.58 4.92
Figure 4.4 Weight distributions for Pacific blue mussels by station, Crofton EEM Cycle Five, March 2009.
20 15 CRO3A (Bisco Island, near-field) 10 5 0 Frequency (n) Frequency 0-1 1-2 2-3 3-4 4-5 5-6 6-7 7-8 Weight (g) 20 15 CRO5A (North Reef, far-field) 10 5 0 Frequency (n) 0-1 1-2 2-3 3-4 4-5 5-6 6-7 7-8 Weight (g) 20 15 CRO4 (Dayman Island, far-far-field) 10 5 0 Frequency (n) Frequency 0-1 1-2 2-3 3-4 4-5 5-6 6-7 7-8 Weight (g)
Crofton EEM Cycle Five 4-17 Hatfield Condition
Mean condition of mussels was similar at far-far-field station CRO4 (103.4 mg/cm3) and near-field station CRO3A (113.6 mg/cm3); at far-field station CRO5A, mussel condition was greater (208.2 mg/cm3) (Figure 4.5).
Figure 4.5 Mean condition (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
250.00
) Mean condition (all stations) 3 200.00
150.00
100.00
50.00 Mean condition index (mg/cm 0.00 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Shell Density
Mean shell density of mussels was similar at all three stations, measuring 1154 mg/cm3 at CRO4 (far-far-field), 1173 mg/cm3 at CRO3A (near-field), and 1222 mg/cm3 at CRO5A (far-field) (Figure 4.6).
Figure 4.6 Mean shell density (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
1400.00 Mean shell density (all stations) ) 3 1200.00
1000.00
800.00
600.00
400.00
200.00 Mean shell density(mg/cm 0.00 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Crofton EEM Cycle Five 4-18 Hatfield Sex Ratios and GSI
At all three stations, relatively more male mussels were present than female mussels (Figure 4.7). The percentage of males versus females was 45% vs. 40% (1.13 ratio) at near-field station CRO3A, 47.5% vs. 42.5% (1.12 ratio) at far-far-field station CRO4, and 57.5% vs. 42.5% (1.35 ratio) at far-field station CRO5A. 10% of the mussels at station CRO4 were immature (i.e., unknown sex), and 15% were immature at station CRO3A. No immature mussels were sampled at station CRO5A.
Mean GSI varied between stations, measuring 8.06 at CRO4 (far-far-field), 12.7 at CRO3A (near-field), and 19 at CRO5A (far-field) (Figure 4.8).
Figure 4.7 Sex ratios for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Male Female Unknow n (Immature)
Figure 4.8 Mean GSI (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
25.0 Mean GSI (all stations)
20.0
15.0
10.0 Mean GSI
5.0
0.0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Crofton EEM Cycle Five 4-19 Hatfield Age Distributions and Size-at-Age
Mean age of mussels varied somewhat between stations. Mussels were overall younger at far-far-field station CRO4 (2.2 years average), while at CRO5A and CRO3A mussels averaged 2.9 and 3 years, respectively (Figure 4.9).
Age frequency distributions were slightly variable between stations (Figure 4.11):
At stations CRO3A and CRO5A, the majority of mussels fell within the 3 year-old age category, with a 2 two year-old individuals; and
At station CRO4, a more wide age distribution was sampled. 2 year-olds were the most prevalent age category, with some 1, 3, and 4 year-olds.
When size-at-age was measured at each station, mussels at CRO5A were larger for their age (89 mg/yr) than mussels at CRO3A (45 mg/yr) and CRO4 (33 mg/yr) (Figure 4.10).
Figure 4.9 Mean age (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
4 Mean age (all stations)
3
2
Mean age (yrs) 1
0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Figure 4.10 Mean size-at-age (±SE) for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
100 90 Mean size-at-age (all stations) 80 70 60 50 40 30 20 10 Mean size-at-age(mg/yr) 0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Crofton EEM Cycle Five 4-20 Hatfield Figure 4.11 Age distributions for Pacific blue mussels by station, Crofton EEM Cycle Five, March 2009.
40 35 CRO3A (Bisco Island, near-field) 30 25 20 15 10 Frequency (n) Frequency 5 0 1234 Age (yr) 40 35 CRO5A (North Reef, far-field) 30 25 20 15 10 Frequency (n) 5 0 1234 Age (yr) 40 35 CRO4 (Dayman Island, far-far-field) 30 25 20 15 10 Frequency (n) Frequency 5 0 1234 Age (yr) ANOVAs
Table 4.6 presents results of ANOVAs performed to evaluate differences in whole organism metrics and indices (i.e., age, total weight, dry meat weight, shell weight, and shell density) among the three sampling stations.
Age, total weight, dry meat weight, and shell weight were all lowest in mussels at reference station CRO4 relative to the other two stations. Dry meat weight was also lower at near-field station CRO3A than at far-field station CRO5A; no difference was identified for age, total weight, and shell weight between near-field and far-field stations.
No statistical differences were observed between stations for shell density.
Crofton EEM Cycle Five 4-21 Hatfield Table 4.6 Tests for differences in mussel whole organism metrics and indices among near-field, far-field, and reference (far-far-field) stations, Crofton EEM Cycle Five, March 2009.
Whole Organism Metrics/Indices ANOVA ANCOVA Type of Slope Intercept Pairwise Comparisons p-value3 Pattern Response Dependent Variable Covariate p-value2 p-value1 p-value2 Ref4 vs. NF Ref4 vs. FF NF vs. FF Survival Age (ranked) - 0.000 --0.000 0.000 0.817 NF, FF > Re f Energy Use Dry meat weight - 0.000 --0.000 0.000 0.000 FF > NF > Ref Total weight (ranked) - 0.000 --0.000 0.000 0.519 NF, FF > Re f
GSI (Log10 mantle weight) Log10 wet meat wt - 0.629 0.006 0.882 0.007 0.004 FF > NF, Ref
Size-at-age (Log10 dry meat weight) Log10 age - 0.493 0.000 0.058 0.000 0.000 FF > NF, Ref
Size-at-age (Log10 total weight) Log10 age - 0.105 0.060 0.254 0.028 0.335 FF > Re f
Energy Storage Condition (Log10 dry meat wt) Log10 total weight - 0.301 0.000 0.028 0.000 0.000 FF > NF > Ref
Condition (Log10 dry meat wt) Log10 cavity volume - 0.914 0.000 0.155 0.000 0.000 FF > NF, Ref Shell weight (ranked) - 0.000 --0.000 0.000 0.135 NF, FF > Re f Shell density (ranked) - 0.409 - - 0.957 0.415 0.577 - 1 The assumption of equal interaction slopes betw een areas w as met (p > 0.01). 2 Bolded value indicates that the means or y-intercepts w ere significantly different betw een areas (p ≤ 0.10). 3 Tukey's comparisons used for ANOVA (p ≤ 0.10); GLM used for ANCOVA after verifying equal interaction slopes betw een station pairs (p ≤ 0.033). 4 Far-far-field used as reference. NF = near-field (CRO3A, Bisco Island), FF = far-field (CRO5A, North Reef), Ref = reference (CRO4, Dayman Island). Table 4.7 Response-based effects assessment for mussels using statistical results, Crofton EEM Cycle Five, March 2009.
Type of Dependent Statistical Reference Exposure % Parameter Covariate (X) Criteria1 Effects?2 Response Variable (Y) Procedure Mean3 Mean3 Difference Survival Age Age None ANOVA NF > Ref Yes (p = 0.000) 36.875 73.175 98% Energy Size Dry Meat Weight None ANOVA NF > or < Ref Yes (p = 0.000) 0.08 0.13 74% Expenditure Total Weight None ANOVA NF > or < Ref Yes (p = 0.000) 37.00 67.85 83% Gonad size Gonad Weight Wet Meat Weight ANCOVA NF > or < Ref No (p = 0.882) -1.24 -1.23 1% Size-at-age Dry Meat Weight Age ANCOVA NF > or < Ref No (p = 0.058) -1.11 -1.01 9% Total Weight Age ANCOVA NF > or < Ref No (p = 0.254) 0.30 0.36 18% Energy Storage Condition Dry Meat Weight Total Weight ANCOVA NF > or < Ref Yes (p = 0.028) -1.11 -1.02 8% Dry Meat Weight Cavity Volume ANCOVA NF > or < Ref No (p = 0.155) -1.09 -1.03 5% Shell Condition Shell Weight None ANOVA NF > or < Ref Yes (p = 0.000) 34.51 65.05 88% Shell Density None ANOVA NF > or < Ref No (p = 0.957) 55.42 57.60 4% 1 Near-field (CRO3A) used as exposure station, far-far-field (CRO4) used as reference. 2 p < 0.10 for ANOVA; p < 0.033 for ANCOVA. 3 Least Squares Mean from ANOVA or ANCOVA test output.
Crofton EEM Cycle Five 4-22 Hatfield ANCOVAs
Table 4.6 presents results of ANCOVAs performed to evaluate differences in whole organism indices (i.e., size-at-age, GSI, and condition) among the three sampling stations.
Condition (using cavity volume), GSI, and size-at-age (using dry meat weight) were lower at the near-field and reference stations relative to the far-field station; no difference was identified between near-field and reference for these variables. Size-at-age (using total body weight) was lower at the far-field station than at the reference station; no difference was identified between the near-field station and either of the other two stations. Condition (using total weight) was lower at the reference station relative to the near-field station, and lower at the near-field station relative to the far-field station.
Response-based Effects Assessment
Statistical results are summarized in Table 4.7 according to criteria for a response-based effects assessment of effluent on mussel populations.
A survival response was observed, whereby mean age in the exposed (near-field) population was greater by 98% relative to reference mussels. This response can be indicative of direct or indirect mortality of oysters or reduced reproduction.
An energy expenditure response was observed, whereby mean size (both according to dry meat weight, and total weight) in the exposed population was greater by 74% and 83%, respectively, relative to reference mussels. This response can be indicative of greater energy use in the exposed population as a result of an increased availability of food resources. Reproductive energy expenditure (i.e., GSI) and size-at-age did not exhibit any response.
An energy storage response was observed, whereby body condition (using body weight as a covariate) and shell condition (according to shell weight) in the exposed population was greater by 8% and 88%, respectively, relative to reference mussels. This response can be indicative of an increased storage of energy in response to effects on the exposure population. Neither of the other indicators of body and shell condition (i.e., condition index using cavity volume as a covariate, and shell density) demonstrated a significant response.
Power Analyses
Table 4.8 presents the results of a post hoc analysis of the statistical power of ANCOVAs to detect a difference (i.e., effect size) of 20 to 30% in mussel condition and GSI between stations, as well an a priori analysis of the number of stations that actually would have been necessary to meet the power requirements (i.e., P≥0.90).
Crofton EEM Cycle Five 4-23 Hatfield Table 4.8 Comparison of Post hoc power analysis of ANCOVAs conducted to test for differences in mussel condition and GSI, and a priori analysis of number of samples actually required to achieve sufficient power, Crofton EEM Cycle Five, March 2009.
Post Hoc Power Analysis A Priori Power Analysis
Whole Actual Actual Actual Required Required Test Organism Sample Sample Effect Actual Post hoc Sample Sample A Priori Groups Indices Size All Size Each Size MSE2 Power (P) Size All Size Each Power (P) 1 Stns (n) Stn (n) (log10) Stns (n) Stn (n)
Condition A. Using total weight as a covariate 20% effect All stations 119 40 0.079 0.019 0.728 193 65 0.900 NF vs Ref 79 40 0.079 0.024 0.728 133 67 0.901 NF vs FF 79 40 0.079 0.019 0.812 106 53 0.902 FF vs Ref 80 40 0.079 0.014 0.907 78 39 0.900 25% effect All stations 119 40 0.097 0.019 0.875 130 44 0.901 NF vs Ref 79 40 0.097 0.024 0.866 89 45 0.900 NF vs FF 79 40 0.097 0.019 0.927 71 36 0.901 FF vs Ref 80 40 0.097 0.014 0.976 53 27 0.902 30% effect All stations 119 40 0.114 0.019 0.953 95 32 0.902 NF vs Ref 79 40 0.114 0.024 0.945 65 33 0.901 NF vs FF 79 40 0.114 0.019 0.977 52 26 0.902 FF vs Ref 80 40 0.114 0.014 0.996 39 20 0.904 B. Using cavity volume as a covariate 20% effect All stations 120 40 0.079 0.015 0.822 153 51 0.901 NF vs Ref 80 40 0.079 0.02 0.799 111 56 0.901 NF vs FF 80 40 0.079 0.014 0.907 78 39 0.900 FF vs Ref 80 40 0.079 0.01 0.969 57 29 0.904 25% effect All stations 120 40 0.097 0.015 0.938 103 35 0.901 NF vs Ref 80 40 0.097 0.02 0.918 75 38 0.902 NF vs FF 80 40 0.097 0.014 0.976 53 27 0.902 FF vs Ref 80 40 0.097 0.01 0.996 38 19 0.900 30% effect All stations 120 40 0.114 0.015 0.984 76 26 0.904 NF vs Ref 80 40 0.114 0.02 0.973 55 28 0.904 NF vs FF 80 40 0.114 0.014 0.996 39 20 0.904 FF vs Ref 80 40 0.114 0.01 1.000 28 14 0.900 GSI A. Using wet meat weight as a covariate 20% effect All stations 110 37 0.079 0.022 0.637 223 75 0.900 NF vs Ref 70 35 0.079 0.028 0.624 155 78 0.901 NF vs FF 74 37 0.079 0.018 0.808 100 50 0.901 FF vs Ref 76 38 0.079 0.021 0.763 117 59 0.902 25% effect All stations 110 37 0.097 0.022 0.797 150 50 0.901 NF vs Ref 70 35 0.097 0.028 0.774 104 52 0.901 NF vs FF 74 37 0.097 0.018 0.924 68 34 0.903 FF vs Ref 76 38 0.097 0.021 0.893 79 40 0.903 30% effect All stations 110 37 0.114 0.022 0.903 109 37 0.900 NF vs Ref 70 35 0.114 0.028 0.880 76 38 0.902 NF vs FF 74 37 0.114 0.018 0.976 49 25 0.900 FF vs Ref 76 38 0.114 0.021 0.960 57 29 0.901
Bold values indicate meets P >= 0.90 pow er objective. 1 Calculated by taking the log10 of 1.2 (20% effect), 1.25 (25% effect), and 1.3 (30% effect). 2 Mean squared error term from ANCOVA model using log10 data. NF = near-field (CRO3A, Bisco Island). FF = far-field (CRO5A, North Reef). Ref = reference (CRO4, Dayman Island).
Crofton EEM Cycle Five 4-24 Hatfield The number of mussels analyzed from each station (i.e., approximately 40) for the Cycle Five control/impact design provided sufficient power to detect a 30% difference in condition and GSI between all three stations (P≥0.90). Sample sizes were also sufficient for detecting a 30% difference in condition and GSI between each pair of sampling stations.
The number of samples was sufficient for detecting a 25% difference in condition using cavity volume as a covariate, was within five samples of being sufficient to detect the same difference in condition using total weight as a covariate, but was not sufficient for detecting a 25% difference in GSI (given the number of immature organisms in the population). 40 samples were not sufficient for detecting a 20% difference in any of the ANCOVA variables.
A priori power analyses based on the current cycle’s effect sizes determined that the optimal number of samples required from each station was 78 to detect a 20% effect, 52 to detect a 25% effect, and 38 to detect a 30% effect.
4.3.2 Oysters 4.3.2.1 Community Metrics
In July 2009, 64-65 oysters were collected and analyzed from each of the three bivalve collection stations (64 at stations CRO5A and CRO4, 65 at station CRO3A). Density of oysters along shorelines at these stations was variable among stations, averaging 45 oysters/m2 at CRO3A (Bisco Island), 12 oysters/m2 at CRO4 (Dayman Island), and 7 oysters/m2 at CRO5A (North Reef) (Figure 4.1).
Figure 4.12 Oyster density at stations sampled for Crofton EEM Cycle Five, July 2009.
50 45 40 ) 2 35 30 25 20 15
Oyster Density (N/m 10 5 0 CRO3A (near-field) CRO5A (far-field) CRO4 (far-far-field) 4.3.2.2 Whole-organism Metrics
Whole-organism metrics are presented in tabular form in Appendix A3 for each Pacific oyster collected at each station. Metrics reported include length, width, girth, whole wet weight, whole volume, wet meat weight, dry meat weight, shell weight, shell volume, cavity volume, and age.
Crofton EEM Cycle Five 4-25 Hatfield Table 4.9 Whole-organism metric summary statistics for Pacific oysters, Crofton EEM Cycle Five, July 2009.
Whole wet Whole Cavity Shell Length Width Gir th Age Meat dry Shell Station Summary Statistic weight volume volume volume (mm) (mm) (mm) (yrs) weight (g) weight (g) (g) (ml) (ml) (ml) CRO3A Sample size (n) 65 65 65 64 65 65 65 65 65 65 Bisco Isl. Minimum 43.0 32.0 19.0 2.0 25.2 1.1 18.1 20.0 10.0 5.0 (near-field) Maximum 129.0 79.0 56.0 9.0 427.7 8.5 342.6 225.0 80.0 200.0 Median 96.0 58.0 42.0 5.0 190.3 4.6 144.9 100.0 40.0 60.0 Mean 93.8 57.9 39.9 4.9 195.8 4.5 153.2 106.7 41.7 65.0 Standard Dev 18.5 10.9 8.9 1.6 96.6 1.8 79.9 53.5 22.1 39.7 Standard Error 2.3 1.4 1.1 0.2 12.0 0.2 9.9 6.6 2.7 4.9 CRO5A Sample size (n) 64 64 64 64 64 64 64 64 64 64 North Reef Minimum 75.0 53.0 19.0 3.0 84.9 2.9 60.3 50.0 20.0 30.0 (far-field) Maximum 190.0 106.0 72.0 10.0 781.2 23.2 642.5 400.0 170.0 310.0 Median 119.5 79.5 45.0 6.0 335.6 12.7 259.9 190.0 65.0 110.0 Mean 121.7 77.4 44.0 5.6 359.6 12.7 283.3 190.9 67.5 123.4 Standard Dev 23.7 11.9 10.0 1.6 164.3 4.3 141.4 78.0 30.0 59.8 Standard Error 3.0 1.5 1.2 0.2 20.5 0.5 17.7 9.7 3.7 7.5 CRO4Sample size (n)64646463646464646464 Dayman Isl. Minimum 80.0 40.0 29.0 3.0 103.1 2.8 24.1 50.0 25.0 20.0 (far-far-field) Maximum 200.0 109.0 75.0 9.0 560.2 23.1 440.0 300.0 150.0 200.0 Median 115.5 77.5 39.5 5.0 260.4 11.6 183.8 150.0 70.0 80.0 Mean 118.9 77.3 41.3 4.9 269.9 12.1 193.0 153.0 71.1 82.0 Standard Dev 24.2 12.2 8.5 1.3 105.5 4.6 87.8 60.0 25.0 40.4 Standard Error 3.0 1.5 1.1 0.2 13.2 0.6 11.0 7.5 3.1 5.1
Crofton EEM Cycle Five 4-26 Hatfield Table 4.10 Whole-organism indices summary statistics for Pacific oysters, Crofton EEM Cycle Five, July 2009.
Condition Index Shell Density Station Summary Statistic Size-at-age (mg/yr) (mg/cm3) (g/cm3) CRO3ASample size (n) 646563 Bisco Island Minimum 52.7 1.29 0.27 (near-field) Maximum 376.7 8.68 2.04 Median 114.3 2.39 0.92 Mean 130.2 2.53 0.95 Standard Dev 61.6 0.91 0.37 Standard Error 7.7 0.11 0.05 CRO5ASample size (n) 646464 North Reef Minimum 72.4 1.48 0.87 (far-field) Maximum 489.1 3.69 5.78 Median 198.5 2.25 2.28 Mean 205.5 2.28 2.33 Standard Dev 75.5 0.37 0.84 Standard Error 9.4 0.05 0.11 CRO4 Sample size (n) 64 64 64 Dayman Island Minimum 42.9 0.24 0.70 (far-far-field) Maximum 468.3 4.39 4.77 Median 176.3 2.46 2.44 Mean 178.0 2.48 2.56 Standard Dev 68.2 0.54 1.03 Standard Error 8.5 0.07 0.13 4.3.2.3 Whole-organism Indices
Whole-organism indices are presented in tabular form in Appendix A3 for each Pacific oyster collected at each station. Indices reported include condition index, shell density, and size-at-age. 4.3.2.4 Summary Statistics
Descriptive statistics summarizing characteristics measured in Pacific oysters from each sampling station are presented in Table 4.9 and Table 4.10.
Size and Weight Distributions
Overall, oysters were largest and heaviest at station CRO5A (far-field), and smallest and lightest at station CRO3A (near-field) (Figure 4.13 and Figure 4.14). Weight frequency distributions for oysters were slightly variable from station to station (Figure 4.15):
Oysters from CRO3A exhibited a relatively normal, but slightly bimodal, weight frequency distribution curve; the highest frequencies of oysters fell within the 150-200 g and, to a lesser extent, the 250-300 g categories; Oysters from CRO5A exhibited a relatively normal weight frequency distribution curve; and Oysters from CRO4 exhibited a slightly positively skewed weight frequency distribution curve; frequencies of smaller oysters were higher than mid- to larger-sized oysters.
Crofton EEM Cycle Five 4-27 Hatfield Figure 4.13 Mean length, width and girth (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009.
Length (mm) Mean length (all stations) 140.0 Width (mm) Mean w idth (all stations) Gir th ( mm) Mean girth (all stations) 120.0
100.0
80.0
60.0 Size (mm) Size
40.0
20.0
0.0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Figure 4.14 Mean whole wet weight (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009.
400 Mean w eight (all stations) 350
300
250
200
150 100 Mean wet weight (g) weight wet Mean 50 0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Crofton EEM Cycle Five 4-28 Hatfield Figure 4.15 Weight distributions for Pacific oysters by station, Crofton EEM Cycle Five, July 2009.
16 CRO3A (Bisco Island, near-field) 12 8 4 0 Frequency (n) 0-50 50-100 100-150 150-200 200-250 250-300 300-350 350-400 400-450 450-500 500-550 550-600 600-650 Weight (g) 16 CRO5A (North Reef, far-field) 12 8 4 0 Frequency (n) 0-50 50-100 100-150 150-200 200-250 250-300 300-350 350-400 400-450 450-500 500-550 550-600 600-650 Weight (g)
16 CRO4 (Dayman Island, far-far-field) 12 8 4 0 Frequency (n) 0-50 50-100 100-150 150-200 200-250 250-300 300-350 350-400 400-450 450-500 500-550 550-600 600-650 Weight (g) Condition
Mean condition of oysters ranged between 158.8 mg/cm3 (CRO3A, near-field) and 205.5 mg/cm3 (CRO5A, far-field); condition of oysters at far-far-field station CRO4 (176.9 mg/cm3) was similar to the mean condition for all three stations (180.4 mg/cm3) (Figure 4.16).
Figure 4.16 Mean condition (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009.
) 250 3 Mean condition (all stations)
200
150
100
50
Mean condition index (mg/cm index condition Mean 0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field)
Crofton EEM Cycle Five 4-29 Hatfield Shell Density
Mean shell density of oysters was similar at all three stations, ranging between 2.3 g/cm3 (CRO5A) and 2.5 g/cm3 (CRO3A and CRO4) (Figure 4.17).
Figure 4.17 Mean shell density (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009.
3.0 Mean shell density (all stations) ) 3 2.5
2.0
1.5
1.0
0.5 Mean shell density (g/cm
0.0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field) Age Distributions and Size-at-Age
Mean age of oysters varied somewhat between stations. Oysters were overall older at far-field station CRO5A (5.6 years average), while oysters averaged 4.9 years at stations CRO3A and CRO4 (Figure 4.18).
Age frequency distributions were similarly variable between stations (Figure 4.19):
At station CRO3A, a range of ages were observed (2 to 9 year-olds). The majority of oysters fell within the 5 year-old age category, with several 3 and 4 year-old individuals;
At station CRO5A, a range of slightly higher ages were observed (3 to 10 year-olds). The majority of oysters fell within the 6 year-old age category, with several 4 and 5 year-old individuals; and
At station CRO4, oysters ranged between 3 and 9 years old. The majority of oysters fell within the 4 year-old age category, with several 5 year-old individuals.
When size-at-age was measured at each station, oysters at far-far-field station CRO4 were slightly larger for their age (2.6 g/yr) than oysters at far-field station CRO5A (2.3 g/yr), and much larger than oysters at near-field station CRO3A (0.95 g/yr) (Figure 4.20).
Crofton EEM Cycle Five 4-30 Hatfield Figure 4.18 Mean age (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009.
7 Mean age (all stations) 6
5
4
3
Mean age(yrs) 2
1
0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field) Figure 4.19 Age distributions for Pacific oysters by station, Crofton EEM Cycle Five, July 2009.
25 CRO3A (Bisco Island, near-field) 20
15
10
Frequency (n) 5
0 2345678910 Age (yr) 25 CRO5A (North Reef, far-field) 20
15
10
Frequency (n) Frequency 5
0 2345678910 Age (yr) 25 CRO4 (Dayman Island, far-far-field) 20
15
10
Frequency (n) Frequency 5
0 2345678910 Age (yr)
Crofton EEM Cycle Five 4-31 Hatfield Figure 4.20 Mean size-at-age (±SE) for Pacific oysters, Crofton EEM Cycle Five, July 2009.
3.0 Mean size-at-age (all stations) 2.5
2.0
1.5
1.0
0.5 Mean size-at-age (mg/yr) size-at-age Mean
0.0 CRO3A CRO5A CRO4 (near-field) (far-field) (far-far-field) ANOVAs
Table 4.11 presents results of ANOVAs performed to evaluate differences in whole organism metrics and indices (i.e., age, total weight, dry meat weight, shell weight, and shell density) among the three sampling stations.
Total weight, dry meat weight, and shell weight were all lowest in oysters at station CRO3A (Bisco Island, near-field) relative to the other two stations. Total weight and shell weight were also lower at station CRO4 (Dayman Island, reference) than at station CRO5A (North Reef, far-field). Dry meat weights were statistically similar between the reference and far-field. Shell density was higher at the near-field station than the far-field station, and statistically similar between near-field and reference.
Oysters in the far-field were statistically older in age than at the near-field and reference stations; ages between near-field and reference were statistically similar.
ANCOVAs
Table 4.11 presents results of ANCOVAs performed to evaluate differences in whole organism indices (i.e., size-at-age and condition index) among the three sampling stations.
Oyster condition (using both cavity volume and total body weight as covariates) and size-at-age (using both total body weight and dry meat weight as dependent variables) was significantly different between stations. Condition and size-at-age were significantly lower in oysters at station CRO3A (Bisco Island, near-field) relative to both far-field and reference stations; in addition, condition according to cavity volume was also significantly lower in reference versus far-field oysters.
Crofton EEM Cycle Five 4-32 Hatfield Table 4.11 Tests for differences in oyster whole organism metrics and indices among near-field, far-field, and reference (far-far-field) stations, Crofton EEM Cycle Five, July 2009.
Whole Organism Metrics/Indices ANOVA ANCOVA 3 Type of Slope Intercept Pairwise Comparisons p-value Pattern Response Dependent Variable Covariate p-value2 p-value1 p-value2 Ref4 vs. NF Ref4 vs. FF NF vs. FF Survival Age - 0.009 --0.9980.019 0.022 FF > Ref, NF Energy Use Total weight (ranked) - 0.000 --0.001 0.002 0.000 FF > Ref > NF Dry meat weight (ranked) - 0.000 --0.000 0.588 0.000 FF, Ref > NF
Size-at-age (Log10 total weight) Log10 age -0.1250.000 0.000 0.102 0.000 FF, Ref > NF
Size-at-age (Log10 dry meat weight) Log10 age 0.426 0.000 0.000 0.879 0.000 FF, Ref > NF
Energy Storage Condition (Log10 dry meat wt) Log10 cavity volume -0.5170.000 0.000 0.050 0.000 FF > Ref > NF
Condition (Log10 dry meat wt) Log10 total weight -0.1430.000 0.000 0.066 0.000 FF, Ref > NF Shell Condition
Shell weight (Log10)-0.000--0.009 0.001 0.000 FF > Ref > NF Shell density (ranked) - 0.002 - - 0.846 0.003 0.015 NF, Ref > FF 1 The assumption of equal interaction slopes betw een areas w as met (p > 0.01). 2 Bolded value indicates that the means or y-intercepts w ere significantly different betw een areas (p ≤ 0.10). 3 Tukey's comparisons used for ANOVA (p ≤ 0.10); GLM used for ANCOVA after verifying equal interaction slopes betw een station pairs (p ≤ 0.033). 4 Far-far-field used as reference. NF = near-field (CRO3A, Bisco Island), FF = far-field (CRO5A, North Reef), Ref = reference (CRO4, Dayman Island). Table 4.12 Response-based effects assessment for oysters using statistical results, Crofton EEM Cycle Five, July 2009.
Type of Dependent Statistical Reference Exposure % Parameter Covariate (X) Criteria1 Effects?2 Response Variable (Y) Procedure Mean3 Mean3 Difference Survival Age Age None ANOVA NF > Ref No (p=0.998) 4.91 4.92 0% Energy Size Dry Meat Weight None ANOVA NF > or < Ref Yes (p = 0.000) 121.88 41.52 -66% Expenditure Total Weight None ANOVA NF > or < Ref Yes (p = 0.001) 98.25 65.55 -33% Size-at-age Dry Meat Weight Age ANCOVA NF > or < Ref Yes (p = 0.000) 1.04 0.64 -39% Total Weight Age ANCOVA NF > or < Ref Yes (p = 0.000) 2.40 2.22 -7% Energy Storage Condition Dry Meat Weight Total Weight ANCOVA NF > or < Ref Yes (p = 0.000) 0.99 0.68 -31% Dry Meat Weight Cavity Volume ANCOVA NF > or < Ref Yes (p = 0.000) 0.96 0.70 -27% Shell Condition Shell Weight None ANOVA NF > or < Ref Yes (p = 0.053) 2.24 2.11 -6% Shell Density None ANOVA NF > or < Ref No (p = 0.846) 109.34 104.08 -5% 1 Near-field (CRO3A) used as exposure station, far-far-field (CRO4) used as reference. 2 p < 0.10 for ANOVA; p < 0.033 for ANCOVA. 3 Least Squares Mean from ANOVA or ANCOVA test output.
Crofton EEM Cycle Five 4-33 Hatfield Response-based Effects Assessment
Statistical results are summarized in Table 4.12 according to criteria for a response-based effects assessment of effluent on oyster populations.
A survival response was not observed; age in the exposed (near-field) population was not different (i.e., 0% difference) relative to reference mussels. This may indicate there has not been a significant direct or indirect mortality of oysters, nor reduced reproduction.
An energy expenditure response was observed, whereby mean size (both according to dry meat weight and total weight) in the exposed population was lower by 66% and 33%, respectively, relative to reference oysters. Similarly, size-at-age (both according to dry meat weight and total weight) in the exposed population was lower by 39% and 7%, respectively. This response may be indicative of lower energy use in the exposed population, likely as a result of limited food availability relative to the elevated numbers (i.e., densities) of oysters clustered together at the near-field station.
An energy storage response was observed, whereby condition (both according to total weight and cavity volume) and shell condition (according to shell weight) in the exposed population were lower relative to reference oysters by 31%, 27%, and 6%, respectively. This response can be indicative of a decreased storage of energy in response to effects on the exposure population. Shell density as a measure of shell condition did not demonstrate a significant response.
Power Analyses
Table 4.13 presents the results of a post hoc analysis of the statistical power of ANCOVAs to detect a difference (i.e., effect size) of 20 to 30% in oyster condition between stations, based on a power objective of P≥0.90.
The number of oysters analyzed from each station (i.e., 64) for the Cycle Five control/impact design provided sufficient power to detect a 20% to 30% difference in condition (using both total weight and cavity volume as covariates) between all three stations (P≥0.90 in all cases).
Sample sizes were also sufficient for detecting a 20% to 30% difference in condition between each pair of sampling stations.
Crofton EEM Cycle Five 4-34 Hatfield Table 4.13 Results of post hoc power analysis of ANCOVAs conducted to test for differences in oyster condition, Crofton EEM Cycle Five, July 2009.
Actual Actual Whole Organism Actual Effect Post hoc Test Groups Sample Size Sample Size Actual MSE2 Indices Size (log )1 Power (P) All Stns (n) Each Stn (n) 10 Condition A. Using total weight as a covariate 20% effect All stations 192 64 0.079 0.015 0.952 NF vs Ref 128 64 0.079 0.015 0.977 NF vs FF 128 64 0.079 0.012 0.992 FF vs Ref 128 64 0.079 0.016 0.970 25% effect All stations 192 64 0.097 0.015 0.993 NF vs Ref 128 64 0.097 0.015 0.997 NF vs FF 128 64 0.097 0.012 1.000 FF vs Ref 128 64 0.097 0.016 0.996 30% effect All stations 192 64 0.114 0.015 0.999 NF vs Ref 128 64 0.114 0.015 1.000 NF vs FF 128 64 0.114 0.012 1.000 FF vs Ref 128 64 0.114 0.016 1.000 B. Using cavity volume as a covariate 20% effect All stations 192 64 0.079 0.02 0.884 NF vs Ref 128 64 0.079 0.021 0.924 NF vs FF 128 64 0.079 0.017 0.962 FF vs Ref 128 64 0.079 0.021 0.924 25% effect All stations 192 64 0.097 0.02 0.970 NF vs Ref 128 64 0.097 0.021 0.983 NF vs FF 128 64 0.097 0.017 0.994 FF vs Ref 128 64 0.097 0.021 0.983 30% effect All stations 192 64 0.114 0.02 0.994 NF vs Ref 128 64 0.114 0.021 0.997 NF vs FF 128 64 0.114 0.017 0.999 FF vs Ref 128 64 0.114 0.021 0.997 Bold values indicate meets P >= 0.90 pow er objective 1 Calculated by taking the log10 of 1.2 (20% effect), 1.25 (25% effect), and 1.3 (30% effect). 2 Mean squared error term from ANCOVA model using log10 data. NF = near-field (CRO3A, Bisco Island) FF = far-field (CRO5A, North Reef) Ref = reference (CRO4, Dayman Island) 4.3.3 Supporting Environmental Variables 4.3.3.1 Habitat Characteristics and Water Quality Habitat characteristics and water quality at bivalve collection stations sampled in 2009 are summarized in Table 4.14 and Table 4.15. Mussels were collected during the spring from bedrock habitat on slopes ranging between 10 and 18%, from a range of aspects (Table 4.14). Water temperature at the three stations was similar, and was highest at the south-facing sampling location. Salinity at far-far-field station CRO4 (27.0 ppt) was lower than at the other two stations, indicating the presence of fresher water; correspondingly, dissolved oxygen was highest at this station (14.3 mg/L). Salinity was closer to 30 ppt at stations CRO3A and CRO5A; correspondingly, dissolved oxygen was relatively lower at these stations (8.49 and 10.3 mg/L, respectively).
Crofton EEM Cycle Five 4-35 Hatfield Table 4.14 Habitat characteristics and water quality at Pacific blue mussel sampling stations, Crofton EEM Cycle Five, March 2009.
Primary Sampling station Aspect Slope (%) Temp (°C) Salinity (ppt) DO (mg/L) Substrate CRO3A (Bisco Isl., near-field) E 10 Bedrock 7.4 29.6 8.49 CRO5A (North Reef, far-field) W 18 Bedrock 7.0 29.5 10.30 CRO4 (Dayman Isl., far-far-field) S 13 Bedrock 7.6 27.0 14.30
Oysters were collected during the summer from sand/gravel/cobble habitats, on slopes ranging between 5 and 5.5%, from north and south aspects (Table 4.15). Salinity was relatively similar between the three stations (26.7 to 26.9 ppt). Water temperature at the three stations varied relative to the sampling aspect; north-facing sampling locations measured 16.8 and 17.6 degrees, and the south-facing sampling location measured 19.2 degrees. Dissolved oxygen also varied between stations, ranging between 7.8 and 11.4 mg/L, but did not appear to vary according to salinity. Similar to water quality measured during the spring mussel sampling program, the highest dissolved oxygen was observed at station CRO4 (Dayman Island) in the far-far-field.
Table 4.15 Habitat characteristics and water quality at Pacific oyster sampling stations, Crofton EEM Cycle Five, July 2009.
Sampling station Slope Primary Underlying Temp Salinity DO Aspect (%) Substrate Substrate (°C) (ppt) (mg/L) CRO3A (Bisco Isl., near-field) N 5.5 Gravel Sand, cobble, bedrock 16.8 26.7 9.87 CRO5A (North Reef, far-field) S/N 5 Gravel Sand, bedrock 17.61 26.91 7.81 CRO4 (Dayman Isl., far-far-field) S 5 Sand Cobble, gravel, bedrock 19.2 26.7 11.40 1 Water quality measured along northern aspect shoreline. 4.3.3.2 Thermographs
Near-shore water temperatures measured at bivalve collection stations between mid-March and early to mid-July 2009 are presented graphically in Figure 4.21. Temperatures at the near-field (CRO3A) and far-field (CRO5A) stations were similar. Temperatures were overall higher at far-far-field station CRO4, likely due to daily exposure of the thermograph to air at the low-low tide as evidenced by sudden high temperatures and large fluctuations occurring during within 2-3 hours of the lowest-low tide. Exposure was likely a result of deploying the thermograph too close to shore within a shallow area just above the low-low tide mark. At the other two stations, evidence of exposure was limited to a handful of days throughout the study period, indicating better positioning of the thermographs.
Temperatures at all stations steadily rose following deployment in March 2009, peaked in mid-June, and remained high until retrieval in early to mid-July.
Crofton EEM Cycle Five 4-36 Hatfield Figure 4.21 Near-shore water temperatures at bivalve sampling stations, Crofton EEM Cycle Five, March to July 2009.
50 45 CRO3A (Bisco Island) 40 24 h moving average 35 30 25 20 15 10 Water temperature (°C) temperature Water 5 0 17-Mar-09 6-Apr-09 26-Apr-09 16-May-09 5-Jun-09 25-Jun-09 15-Jul-09
50 CRO5A (North Reef) 45 24 h moving average 40 35 30 25 20 15 10 Water temperatureWater (°C) 5 0 17-Mar-09 6-Apr-09 26-Apr-09 16-May-09 5-Jun-09 25-Jun-09 15-Jul-09
50 45 CRO4 (Dayman Island) 40 24 h moving average 35 30 25 20 15 10 Water temperature (°C) 5 0 17-Mar-09 6-Apr-09 26-Apr-09 16-May-09 5-Jun-09 25-Jun-09 15-Jul-09 Date
Crofton EEM Cycle Five 4-37 Hatfield 4.4 DISCUSSION
Historical studies in the vicinity of the Crofton pulpmill from the 1960s to the early 1980s (e.g., Quayle 1969, Anderson 1977, Ellis et al. 1981) reported effects of effluent on condition of local oysters. Since this time, considerable improvements in effluent quality have occurred at the mill, including improved water and fibre control, elimination of elemental chlorine during the bleaching process, and implementation of secondary effluent treatment in 1992 (Hatfield 1994). Similarly, results of oyster surveys performed in Cycles One, Two, and Three also indicated that oyster populations were no longer similarly affected by pulpmill effluent exposure.
In Cycle Five, both mussels and oysters were surveyed to determine the current state of effluent effects on fish near Crofton.
Mussels at the reference station were significantly younger, smaller, and lighter, and were present in higher densities relative to those at the near-field and far field stations. Lower densities of older, larger, heavier mussels were present closer to the mill, indicating greater survival and energy use in these organisms. Some indicators of body and shell condition, representative of energy storage, were also significantly greater in mussels closer to the mill, both in the near-field and far-field, relative to the reference area. Reproductive energy use represented by GSI and energy use represented by size-at-age did not show any significant differences between near-field and reference mussels, although these parameters were significantly higher in far-field mussels.
In contrast to mussels near the Crofton mill which demonstrated greater survival, energy use, and energy storage than those in the reference area, near-field oysters demonstrated lower survival, energy use, and energy storage than reference oysters. Oysters at the near-field station were significantly smaller and lighter, both in general and for their age, and were present in higher densities relative to those at the far-field and reference stations. Indicators of body condition and shell condition were also significantly lower in oysters closer to the mill relative to the other two stations. Overall, the characteristics of oysters at the near-field station indicated limited food availability relative to the elevated numbers (i.e., densities) of organisms clustered tightly together.
More male mussels than female mussels were present at all three stations, although ratios were close to 1:1. Ratios of male versus female mussels were similar between the near-field (1.13) and reference (1.12) areas, and GSI did not demonstrate any significant difference between the two areas, indicating that sex did not play a role in influencing the significantly greater survival, energy use, and energy storage of mussels in the near-field. Where younger mussels (ages 1 to 2) were found at the reference site, mussels in the near-field were older and no 1 year-olds were collected. This would indicate that recent spawning and/or spat settlement was not successful at the near-field station.
Crofton EEM Cycle Five 4-38 Hatfield On average, oysters in the far-field were older, and mean ages of oysters in the near-field and the reference area were similar. In contrast to mussels, the youngest oysters in the study (2 year-olds) were only found at the near-field station, and in terms of their size-at-age the near-field oysters were substantially smaller.
Coexistence of mussels and oysters can be difficult, with the larger oysters often colonizing suitable habitat at the expense of mussels (Diederich 2005). Temperature and dissolved oxygen were higher at the reference site where oysters were larger and heavier, and mussels were smaller. Pacific oysters are known to more rapidly colonize in more favourable water quality conditions characterized by higher temperatures and dissolved oxygen levels (Cardaso et. al 2007), so it is possible that oyster populations have settled at the reference station at the expense of mussels.
In previous cycles, proximity to the mill was not found to have significant effects on oyster reproduction, recruitment, development, or adult condition of Pacific oysters. In Cycle Five, however, the response-based effects summary demonstrated significant effects between the exposure and reference areas for both mussels and oysters. Given that opposing effects were observed for the two species, it is difficult to say whether the differences are associated with natural or mill-related factors.
Crofton EEM Cycle Five 4-39 Hatfield 5.0 FISH TISSUE SURVEY
Summary of Fish Tissue Survey for Crofton EEM Cycle Five: Chlorinated dioxins and furans were monitored in sediments and crab within the receiving environment during Cycle Five as a requirement by Health Canada and Environment Canada (i.e., separate from EEM); results of the most recent monitoring program (i.e., 2009) are summarized herein. In 2009, total TEQ concentrations in crabs at four stations were below the guideline (24.4 pg/g hepatopancreas), and levels at the remaining three stations had declined relative to 2006; and A fish tainting survey was not required for Cycle Five, given that no reports related to Crofton effluent were received by public officials in recent years.
Detailed results and analyses for the 2009 dioxin/furan trend monitoring program at Crofton are presented in Hatfield (2009b); a summary is presented below.
5.1 TISSUE ANALYSES: DIOXINS AND FURANS
Based on ongoing recreational consumption advisories in Stuart Channel (Figure 5.1), the Crofton mill was required to monitor dioxins and furans in crab and sediments during the Cycle Five EEM Program, as per Government of Canada Directives.
5.1.1 Sediment
Government of Canada Directives required sampling at station S6, near the Crofton outfalls. Loss on ignition (LOI), an indicator of organic content, has generally declined near the mill in association with a decreased level of finer sediments in the area. In 2009, LOI was 7.5% at S6 and sediments in this area were predominantly sandy (71%), with some mud (29%).
Sediment organochlorine concentrations have generally decreased at station S6 since monitoring began in 1990 (Figure 5.2). Concentrations of 2,3,7,8-T4CDD, 2,3,7,8-T4CDF, and Total TEQ in 2009 were all within the lower end of the range of concentrations observed in previous years at the station (2.29 pg/g, 60.7 pg/g, and 36.9 pg/g, respectively).
5.1.2 Crab Hepatopancreas
Federal directives required crab collections at 8 stations in Stuart Channel: C1 (Lamalchi Bay), C2 (Willy Island), C3 (Outfalls), C4 (Dock Bay), C7 (Maple Bay), C8 (Burgoyne Bay), C17 (Boulder Point), and C23 (Dayman Island). Required numbers of adult male Dungeness crab (n=7) were captured at stations C2, C3, and C8. Target numbers of crab could not be collected at the other five stations: five crabs were caught at C4, four crabs at C17, and two crabs at C1 and C7; none were collected at C23.
Crofton EEM Cycle Five 5-1 Hatfield Figure 5.1 Fisheries closures in the vicinity of Catalyst Paper, Crofton Division, 2009.
123°45'W 123°30'W
S t r
a 49°0'N i Thetis t
49°0'N Island o f
T G r e i o n r Kuper c g South o i n m a Ladysmith Island o t s a Harbour S s s l u a t i o u P C 40 a H h a r t n n e l
Galiano
Island Chemainus R. C h a n n Catalyst Paper e Corporation l Crofton Division Saltspring Island
Maple Bay 125 Burgoyne Bay 60 Cowichan R.
Cowichan 48°45'N Bay 48°45'N
123°45'W 123°30'W
LEGEND Closed to commercial crab fishing with a health advisory on non-commercial crab fisheries on the consumption of crab hepatopancreas
# Crab hepatopancreas consumption limit (grams/week) 0482 Km Scale 1:250,000 Projection: Albers Equal Area - NAD83 t K:\Data\Project\CR1327\GIS\_MXD\B_InterpretiveReport\CR1327_B01_2009Closure_20100112.mxd In 2009, concentrations of 2,3,7,8-T4CDD in crab hepatopancreas ranged from 0.94 pg/g (C17) to 2.59 pg/g (C4). 2009 concentrations of 2,3,7,8-T4CDD in crabs were lower than concentrations measured in 2006 (Cycle Four) at all stations (Figure 5.3). At three stations (C1, C7, and C17), crab dioxin concentrations in 2009 were the lowest values recorded since the onset of monitoring. At the remaining stations (C2, C3, C4, and C8), concentrations were within the lower range of those reported in recent years, and considerably lower than concentrations reported in the early to mid 1990s, when concentrations were as high as 20 pg/g (C3).
2009 concentrations of 2,3,7,8-T4CDF in crab hepatopancreas ranged between 21 pg/g (C17) and 55.9 pg/g (C3). Similar to dioxins, concentrations of 2,3,7,8-T4CDF in crabs were lower in 2009 than concentrations measured in 2006 (Cycle Four) at all stations (Figure 5.4). At four stations (C1, C4, C8, and C17), crab furan concentrations in 2009 were the lowest values recorded since the onset of monitoring. At the remaining stations (C2, C3, and C7), concentrations were within the lower range of those reported in recent years, and considerably lower than concentrations reported in the early to mid 1990s, when concentrations were as high as 670 pg/g (C3).
Crab total TEQ concentrations in 2009, ordered from largest to smallest, were: 37.0 pg/g at C8, 34.7 pg/g at C4, 28.2 pg/g at C3, 22.1 pg/g at C7, 20.9 pg/g at C2, 18.3 pg/g at C1, and 10.6 pg/g at C17. TEQ concentrations in crab hepatopancreas from three stations exceeded Health Canada’s consumption advisory criteria of 24.4 pg/g TEQ: C3 (by 1.2%), C4 (by 1.4%), and C8 (by 1.5%). TEQ concentrations at all other stations were below the guideline (Figure 5.5). At station C1, 2009 was the first year TEQ concentrations in crab hepatopancreas fell below the consumption guideline. Overall, 2009 TEQ concentrations did not appear to vary consistently with distance from the mill.
Total TEQ concentrations in crabs have declined considerably since the onset of organochlorine monitoring at the mill, given the gradual phasing out of elemental chlorine bleaching to its complete elimination in 1996. Although the rate of decline has slowed, concentrations continue to drop and have not yet leveled off. At three stations (C1, C4, and C17), crab TEQ concentrations in 2009 were the lowest values recorded since the onset of monitoring. At the remaining stations (C2, C3, C7, and C8), concentrations were within the lower range of those reported in recent years, and considerably lower than concentrations reported in the early to mid 1990s, when concentrations were as high as 186 pg/g (C4).
5.2 TAINTING EVALUATION
No reports of fish tainting related to Crofton effluent have been received by public officials in recent years; therefore, tainting evaluations were not conducted for Cycle Five.
Crofton EEM Cycle Five 5-3 Hatfield Figure 5.2 Crofton sediment 2,3,7,8-T4CDD, T4CDF and TEQ, 1990 to 2009.
123°45'W 123°30'W
2,3,7,8-T4CDD 2,3,7,8-T4CDF
16 15.00 700 14 600 590 12 S500 10.00 10 9.05 t r
7.90 400 340 a 49°0'N
8 i 295 Thetis t 270 5.50 300 5.18 6 5.10 49°0'N 4.30 4.10 173 Island 170 3.70 3.57 200 o
130 120 120
4 2.60 f 111 110 2.29 2.12 2.00 pg/g (dry weight)
pg/g (dry weight) 71 60 61 54 100 48 2 0.79 G 18 ND e 0 South Kuper 0 o r Ladysmith Island n g o t s i s s 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Harbour1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 a
a Year u Year o P
H
S t T u r a i n r c t o m Galiano Diffuser a Island !( l S6 i C C Chemainus R. h h
Osborn a a n Booth n Bay n n Bay e Catalyst Paper e l Corporation l Crofton Division Saltspring Island
Maple Total TEQ (2005 TEFs) Bay
250 237.9 Burgoyne 200 Bay 155.5 152.4 Cowichan R. 150 134.8 48°45'N 94.3 100 89.8 75.1 70.3 67.6 61.0 60.0 48°45'N 40.3 36.9 35.0
pg/g (dry weight) 50 27.3 16.2 12.5 0 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year
123°45'W 123°30'W
LEGEND
!( 2009 Sediment Site
ND Not Detected
0512.5 0 Km Scale 1:250,000 Projection: Albers Equal Area - NAD83 t
K:\Data\Project\CR1327\GIS\_MXD\B_InterpretiveReport\CR1327_B04_SedResults_20100112.mxd Figure 5.3 Crofton Dungeness crab hepatopancreas 2,3,7,8-T4CDD, 1990 to 2009.
C2 20 C1 20 18 18 16 16 16.0 14 14.0 14 12 12 12.0 10 10 8.3 8 7.9 8 6
6 4.8 3.2 pg/g (dry weight) pg/g (dryweight)
4 2.9 2.9 2.9 2.7 4 2.1 2.0 2.1 1.7 1.6 2 2 1.3 NC NC NC NC NC NC NC 0 0 NC NC NC NC NC NC NC NC NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
20 20.0 C3 20 C4 18 18
16 16 15.5 14.0
14 13.0 14 12.0
12 12 11.0 10.4 10.0 10 10.0 10 8 8.2 8 6.1 5.6
6 5.0
4.6 6 4.6 4.4 4.4 3.9 4.0 3.5 3.6 pg/g (dry weight) 3.2 pg/g (dry weight) 3.2 3.0 3.1
4 2.8
2.6 4 2.7 2.3 2.6 2.4 2.1 2.0 2 1.9 2 0 0 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
20 C7 20 C8 18 18 16.0
16 15.0 16 14 14 11.0 12 12 11.0 10 9.8 10
8 8 7.7 6.5 6.4 5.3 5.2 4.8 4.8 6 6 5.0 4.4 4.2 4.0 3.9 3.3 pg/g (dryweight) 3.4 3.1 pg/g (dry weight) 4 3.2 2.5 4 2.8 2.4 2.4 2.5 1.4 1.5
2 2 1.0 NDR NDR NC 0 0 NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
20 C17 20 C23 18 18 16 16 14 14 12 12 10 10
8 7.1 8 6 6 pg/g (dryweight) pg/g (dryweight) 4 3.2 4 2.4 2.4 1.9 1.8 1.7 1.6 1.5 1.4 1.4 1.0 2 0.9 2 NC NC NC NC NC NC NC NC NC 0 0 NC NC NC NC NC NC NC NC NC NC NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
Figure 5.4 Crofton Dungeness crab hepatopancreas 2,3,7,8-T4CDF, 1990 to 2009.
C2 700 C1 700 600 600 500 500 430.0
400 400 360.0 350.0
300 260.0 300
200 190.0 200 pg/g (dry weight) pg/g (dryweight) 95.0 87.0 83.0 75.5 72.1 72.0 56.0 100 60.0
53.3 100 37.7 40.1 31.1 NC NC NC NC NC NC NC 0 0 NC NC NC NC NC NC NC NC NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
C3 700 670.0 700 C4 600 600 520.0 500 500 400.0 400 390.0 400 340.0 310.0 310.0 290.0 290.0 300 300 280.0 190.0
200 160.0 160.0 200 160.0 140.0 140.0 125.0 126.0 110.0 pg/g (dry weight) pg/g (dry weight) 110.0 90.0 97.4 76.0 76.9 73.0 71.0 67.0 77.0 76.5 76.0
100 55.9 100 61.6 41.3 47.7 0 0 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
700 C7 700 C8 600 600
500 490.0 500 400 410.0 400 290.0 280.0
300 260.0 300 190.0 185.0
200 170.0 200 160.0 135.0 130.0 pg/g (dryweight) pg/g (dry weight) 110.0 110.0 99.3 97.0 86.0 88.0 87.0 78.3 82.9 76.0 76.0 73.0 59.0 62.0
100 56.0
51.0 100 52.6 37.3 17.0 9.6 NC 0 0 NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
700 C17 700 C23 600 600 500 500 400 400 300 300
200 160.0 200 pg/g (dryweight) pg/g (dryweight) 82.0 61.9 100 100 64.0 51.0 49.4 44.0 33.5 36.2 28.0 33.0 21.0 22.7 NC NC NC NC NC NC NC NC NC 0 0 NC NC NC NC NC NC NC NC NC NC NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
Figure 5.5 Crofton Dungeness crab hepatopancreas 2,3,7,8-T4CDD TEQs (WHO 2005 TEFs), 1990 to 2009.
C2 700 C1 700 600 600 500 500 430.0
400 400 360.0 350.0
300 260.0 300
200 190.0 200 pg/g (dry weight) pg/g (dryweight) 95.0 87.0 83.0 75.5 72.1 72.0 56.0 100 60.0
53.3 100 37.7 40.1 31.1 NC NC NC NC NC NC NC 0 0 NC NC NC NC NC NC NC NC NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
C3 700 670.0 700 C4 600 600 520.0 500 500 400.0 400 390.0 400 340.0 310.0 310.0 290.0 290.0 300 300 280.0 190.0
200 160.0 160.0 200 160.0 140.0 140.0 125.0 126.0 110.0 pg/g (dry weight) pg/g (dry weight) 110.0 90.0 97.4 76.0 76.9 73.0 71.0 67.0 77.0 76.5 76.0
100 55.9 100 61.6 41.3 47.7 0 0 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
700 C7 700 C8 600 600
500 490.0 500 400 410.0 400 290.0 280.0
300 260.0 300 190.0 185.0
200 170.0 200 160.0 135.0 130.0 pg/g (dryweight) pg/g (dry weight) 110.0 110.0 99.3 97.0 86.0 88.0 87.0 78.3 82.9 76.0 76.0 73.0 59.0 62.0
100 56.0
51.0 100 52.6 37.3 17.0 9.6 NC 0 0 NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
700 C17 700 C23 600 600 500 500 400 400 300 300
200 160.0 200 pg/g (dryweight) pg/g (dryweight) 82.0 61.9 100 100 64.0 51.0 49.4 44.0 33.5 36.2 28.0 33.0 21.0 22.7 NC NC NC NC NC NC NC NC NC 0 0 NC NC NC NC NC NC NC NC NC NC NC NC 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2006 2009 Year Year
6.0 BENTHIC INVERTEBRATE COMMUNITY SURVEY
Summary of Benthic Invertebrate Community Survey for Crofton EEM Cycle Five: The overall objective of the Cycle Five benthic invertebrate community survey was to determine effects of effluent exposure on benthos in the Crofton area. The survey objective was achieved by sampling for benthic invertebrates, supporting sediment and water quality data, along a gradient of twelve stations placed at increasing distances from the mill outfalls. The gradient design was based on those performed in Cycles Two and Three, and made it possible to determine if effects on benthic invertebrates were evident along the exposure gradient; Although conditions have improved since Cycle Three, benthic communities in Cycle Five continued to demonstrate effects along the effluent exposure gradient. Closer to the outfalls, benthic communities exhibited higher densities, were dominated by Capitellid worms, and were statistically less similar to reference communities farther away; Although sediment quality has improved since Cycle Three, closer to the outfalls sediments continued to be characterized by higher C:N, organic carbon, and lower redox potential; Sulphides appeared to be relatively similar between near-field and far-field stations, indicating that oxygen is still available to microbes in sediments throughout the study area; According to the impact grading scheme, conditions since Cycle Three have improved; Cycle Five near-field sediment quality ranged between normal to moderately impacted, and benthic communities were representative of normal to low impact/enrichment conditions; and As in Cycle Three, the Cycle Five benthic study demonstrated that, although the historical fibre mat in the vicinity of the Crofton outfalls has decomposed to a sufficient extent that it no longer exerts a toxic (inhibitory) effect on benthic communities, the remaining organic matter contributes to a mild enrichment effect at stations close to the outfalls.
6.1 INTRODUCTION
A subtidal benthic invertebrate community survey was performed near Crofton in Cycle Five to assess effects with distance from the mill outfalls, predominantly related to historical effluent deposits. The survey was undertaken to satisfy federal Environmental Effects Monitoring (EEM) Cycle Five requirements, and was identical in scope and scale to similar surveys performed during Cycles Two and Three, to enable temporal comparisons to be made. The program examined spatial differences in sediment quality and structural differences in invertebrate communities (i.e. density, number of taxa, shifts in dominance, and diversity) in the vicinity of the mill effluent outfalls.
Similar benthic invertebrate community surveys were undertaken in the vicinity of Crofton during EEM Cycle One (March 1995), Cycle Two (March 1999), Cycle Three (February 2003), and historically. Surveys of sediment quality in the immediate vicinity of the Crofton Division outfalls also were conducted in the early 1990s (see Hatfield 1994). These early surveys indicated the presence of anoxic beds of fibrous, organic material in the vicinity of the diffuser, and impacted benthic communities. At the time, few benthic species were present in the vicinity of the diffusers. The Cycle One survey conducted in 1995 indicated substantial improvement in the near-field area, with similar levels of density and taxonomic richness exhibited relative to far-field stations. The Cycle Two 1999 gradient survey found no significant differences in benthic invertebrate community endpoints along a gradient of effluent exposure.
Crofton EEM Cycle Five 6-1 Hatfield This section reports methods, results, and interpretations for the Cycle Five benthic invertebrate community survey conducted in March 2009. This section follows the reporting guidelines recommended by Environment Canada for Cycle Five interpretive reports (Environment Canada 2005).
6.2 METHODS
6.2.1 Modifications to Sampling Design
No major changes to the sampling design were made relative to the Cycle Five design document (Hatfield 2009a).
6.2.2 Gradient Design
The Cycle One benthic invertebrate survey used a control/impact sampling design; however, a gradient response (i.e., with distance from the mill) was observed in the channel during that cycle. The Pulp and Paper EEM Technical Guidance Document was updated to recommend that a simple gradient design be used for effects assessments in non-homogeneous, narrow estuaries or geographically restricted marine bays, inlets, or fjords (Environment Canada 2005). This type of design involves sampling along a gradient through declining levels of effluent concentration in the receiving environment; far-far-field stations are used as reference stations and are positioned far enough down the gradient that they are presumed to be exposed to little or no effluent. A gradient design was adopted after Cycle One for subsequent invertebrate surveys near Crofton.
The Cycle Five benthic invertebrate community survey was conducted using a gradient design (i.e., stations were placed at increasing distances from the mill outfalls). The design made it possible to determine if a gradient of effects on benthic invertebrates was evident along the exposure gradient. The gradient design applied in this cycle was based on those performed in Cycles Two and Three, to enable historical comparisons.
6.2.3 Sampling Locations and Collection Dates
Benthic invertebrate communities were sampled at 12 stations along the exposure gradient near the Crofton mill from March 10 to 12, 2009 (Figure 6.1). Station locations were identical to those used in Cycle Three. Three replicates were collected from each of the 12 stations in order to evaluate benthic communities, for a total of 36 samples; these three replicates were also examined for sediment quality, and an additional fourth replicate was collected to evaluate sediment grain size (Table 6.1). Efforts were made, where possible, to collect all samples from a similar substrate and target depth range of 40 to 50 m.
Crofton EEM Cycle Five 6-2 Hatfield 6.2.4 Field Sampling Procedures 6.2.4.1 Sampling Platform
Samples were collected by experienced Hatfield personnel from the MV Lobo, a custom-built 7-metre aluminum vessel with dual 4-stroke 115hp Yamaha outboard engines designed specifically for marine sediment and marine habitat related work. The vessel was equipped with a hydraulic winch system, VHF radio, and all safety equipment required by Transport Canada. Station locations were determined using an on-board differentially corrected Global Positioning System (GPS) integrated with digital nautical charts. Depth at each sampling station was recorded from the depth sounder (sonar).
6.2.4.2 Benthic Invertebrate Surveys
Three benthic invertebrate samples were collected at each station using a 20L stainless-steel Van Veen sediment grab sampler (sample area = 0.1 m2). Upon retrieval, the grab was allowed to sit in a plastic tub until all excess water had drained out. The grab doors were then opened, and the surface sediments were photographed and subsampled for supporting sediment chemistry variables (see Section 6.2.4.3).
Crofton EEM Cycle Five 6-3 Hatfield Table 6.1 Benthic invertebrate sampling locations and collections, Crofton EEM Cycle Five, 2009.
Mean 1 Samples Subsamples Submitted Station Collection Date Coordinates Depth Collected for Analyses (m)
CRB5C 10-Mar-2009 123° 41.083' W 54.1 3 benthos/sed 3 jars (1 from each of 3 replicates) (Alarm Rock) 48° 57.493' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB5B 11-Mar-2009 123° 39.895' W 61.0 3 benthos/sed 3 jars (1 from each of 3 replicates) (Escape Reef) 48° 56.188' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB4A 11-Mar-2009 123° 40.411' W 49.7 3 benthos/sed 3 jars (1 from each of 3 replicates) (Willy Island) 48° 54.810' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB5A 10-Mar-2009 123° 37.346' W 49.6 3 benthos/sed 3 jars (1 from each of 3 replicates) (North Reef) 48° 54.557' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB1A 12-Mar-2009 123° 38.364' W 59.7 3 benthos/sed 3 jars (1 from each of 3 replicates) (North of Outfall) 48° 53.699' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB2 11-Mar-2009 123° 38.049' W 68.0 3 benthos/sed 3 jars (1 from each of 3 replicates) (Outfall) 48° 53.557' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB2A 12-Mar-2009 123° 37.776' W 49.7 3 benthos/sed 3 jars (1 from each of 3 replicates) (Indian Reef) 48° 53.351' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB3 11-Mar-2009 123° 37.301' W 50.6 3 benthos/sed 3 jars (1 from each of 3 replicates) (South of Outfall) 48° 52.773' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB7 10-Mar-2009 123° 37.389' W 46.0 3 benthos/sed 3 jars (1 from each of 3 replicates) (Osborn Bay) 48° 52.066' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB6A 12-Mar-2009 123° 35.617' W 49.2 3 benthos/sed 3 jars (1 from each of 3 replicates) (Parminter Pt.) 48° 54.264' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB6B 12-Mar-2009 123° 35.026' W 45.0 3 benthos/sed 3 jars (1 from each of 3 replicates) (Dock Point) 48° 53.191' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS) CRB6 10-Mar-2009 123° 33.910' W 44.3 3 benthos/sed 3 jars (1 from each of 3 replicates) (Booth Bay) 48° 51.252' N quality grabs; for analysis of TOC, TN (ALS) 1 sed grain 1 jar from 4th replicate size grab for analysis of grain size (ALS)
1 Centroid between replicate grab locations; coordinates for individual grabs available from field data sheets.
Crofton EEM Cycle Five 6-4 Hatfield Figure 6.1 Crofton EEM Cycle Five benthic invertebrate sampling locations, March 2009.
123°40'W 123°35'W T r in co m a l i C h a n n e Kuper l
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.! CRB3 Osborne Crofton Bay Booth .! CRB7 Bay Catalyst Paper Corporation Crofton Division .! CRB6
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123°40'W 123°35'W
LEGEND Depth (metres) Waterbody Intertidal 0 - 20 Stream Network 20 - 50 50 - 100 Pulpmill 100 - 150 0120.5 Km 150 - 200 Scale 1:100,000 .! Benthos Sampling Station Projection: Albers Equal Area - NAD83 200 - 250 t
K:\Data\Project\CR1327\GIS\_MXD\B_InterpretiveReport\CR1327_B06_Benthos_20100112.mxd The grab jaws were then opened, releasing the remaining contents of the grab into the tub. Any sample remaining on the grab was carefully rinsed into the tub using ambient seawater. The station ID and replicate number were recorded on the benthic field collection sheet, and on a waterproof label which was placed on the outside of the tub. Tubs were then transported to dockside sieving stations and processed immediately (see Section 6.2.5.1). After sampling at each station, the grab and other stainless steel equipment were scrubbed and rinsed with ambient seawater in order to avoid cross-contamination between stations.
6.2.4.3 Supporting Sediment Quality Variables
Sediment quality was evaluated within each of the three replicates collected at the 12 benthic invertebrate grab stations in order to provide habitat characteristics and aid in the interpretation of benthic invertebrate community data. The following variables were measured from the top 2 cm of sediment in each replicate grab sample:
Total organic carbon;
Total nitrogen;
Redox potential; and
Total sulphides.
Four randomly selected 2 cm x 2 cm subsamples (4 cm2) were collected and composited from the top 2 cm of sediment of each grab sample, placed in a small plastic beaker to create a 16 cm2 composite, and analyzed immediately in the field for redox potential and sulphides by Aquametrix Research (Courtenay BC). Aquametrix performed their analyses according to the BC Ministry of Environment Protocols for Marine Environmental Monitoring (BC MOE 2002). Also consulted in support of the provincial protocols were the Recommended Protocols for Measuring Conventional Sediment Variables in Puget Sound from the U.S. Environmental Protection Agency and the Puget Sound Water Quality Authority (PSEP 1986). Sulphide readings were taken using a ThermoOrion 290A plus pH/ion/mV meter and 9678BNWP probe; redox potential was analyzed using a VWR Symphony SP301 pH/ion/mV meter and a 9616BNWP probe by ThermoOrion. Appendix A9 presents a detailed explanation of chemical preparations, calibration techniques, analysis methods, and QA/QC protocols.
An additional composite of four randomly selected 2 cm x 2 cm subsamples (for a total of 16 cm2) was collected from the same grab and placed in a labelled 125 mL glass jar for analyses of Total Organic Carbon (TOC) and Total Nitrogen (TN).
Subsampling for redox/sulphides and TOC/TN anlayses was repeated for each of the three replicate grabs at each station. A fourth grab sample was then collected at each station to obtain sufficient sediments from the top 2cm to fill a large, labelled Ziploc bag for sediment grain size analyses.
Crofton EEM Cycle Five 6-6 Hatfield All containers and lids were pre-labelled with the appropriate sample ID number using an indelible marker or adhesive sticker. Matching sample IDs were written on the data sheet for each station. Sediments to be analyzed for TOC, TN, and sediment grain size were shipped to ALS Environmental (Vancouver, BC) for analysis (Appendix A5).
6.2.4.4 Supporting Water Quality Variables
The following water quality variables were measured in near-bottom water at each station, to aid in the interpretation of benthic invertebrate data:
Dissolved oxygen; Temperature; Salinity; and Depth.
One near-bottom water samples was collected at each station using a stainless steel Kemerrer bottle. The bottle was brought to the surface and instantaneous measurements of temperature (±0.1°C), salinity (±0.1‰, parts per thousand) and dissolved oxygen (±0.3mg/L) were made using a multi-probe YSI meter (600-series). The YSI meter was calibrated prior to use according to manufacturer's instructions.
6.2.5 Field Processing Procedures 6.2.5.1 Sample Sieving and Preservation
Benthic invertebrate samples were field-sieved on a near-by wharf in Crofton by Biologica Environmental Services (Victoria, BC) using stands equipped with 1.0 mm mesh-screened trays. Small portions of each grab sample were individually washed with a gentle water flow in order to minimize the retention time of organisms on the screens, and reduce specimen damage. In order to prevent the introduction of zooplankton or other organisms into the sample, ambient seawater used to wash the samples was filtered through 250-μm mesh placed over the intake screens. Large, fragile organisms were removed from the screen during sieving to prevent damage, placed in small externally labelled vials, and preserved with ethanol.
A funnel was used to direct the washed sample fractions into 1-L plastic jars in order to catch all the debris retained on the screens. Sample jars were labelled, and samples requiring more than one container were labelled in series. Excess seawater from each 1-L jar was drained through a 63-μm mesh; if any material was retained on the mesh, it was then washed back into the sample jar. Fifty millilitres of full strength buffered formalin was added to each 1-L jar as a preservative, and mixed into the debris by gently rolling the sample jars. Material that was too large to fit into a 1-L jar (e.g., large pieces of wood) was placed in large, labelled, zippered storage bags with an appropriate amount of preservative. Vials containing the more fragile organisms were placed in the appropriate jar along with the rest of the sample.
Crofton EEM Cycle Five 6-7 Hatfield A field log was kept to record sample IDs, collection dates, washing times, observations, number of jars, and name of the washing technician responsible for each sample. Photographs were taken of each of the samples while washing was in progress, and a debris description was prepared following sorting.
Following sieving, samples remained preserved with formalin for 48 hours for immediate shipment to Sandy Lipovsky (Columbia Science, Royston BC), an expert invertebrate taxonomist who is familiar with marine benthos in British Columbia. Upon receiving the samples, Columbia Science transferred them to ethanol prior to sorting for identification.
6.2.6 Laboratory Procedures 6.2.6.1 Taxonomic Analysis
Field-sieved and preserved benthic samples were hand-delivered to Columbia Science for sorting and identification. Invertebrates were identified to species or the lowest practicable taxonomic level, as recommended in Table 4-6 in the TGD (Environment Canada 2005).
6.2.6.2 QA/QC and Verifications
As per the TGD, the following QA/QC procedures were used during laboratory analysis of benthic invertebrates:
All personnel involved in sample processing and analysis were appropriately trained in handling and identifying marine benthic invertebrates.
Four samples were split in advance of analyzing remaining samples to test if splitting (i.e., subsampling) was a reliable option for estimating invertebrate communities in larger samples containing finer substrate.
All samples were subjected to a 20%, 50%, or 100% re-sort to determine sorting error; if sorting error exceeded 10% (i.e., sorting efficiency < 90%), a complete re-sort of that sample was required.
Appropriate taxonomic references were used for the study area location and habitat.
New taxa identified during Cycle Five that had not been identified during previous cycles were verified by taxonomists at Biologica Environmental Services for inclusion in the reference collection.
Crofton EEM Cycle Five 6-8 Hatfield 6.2.7 Analytical Approach 6.2.7.1 Data Handling
Benthic invertebrate data were entered into an electronic spreadsheet by Columbia Science, who checked for transcription errors. Data were organized into “adult”, “intermediate”, and “juvenile” classifications for Cycle Five by the taxonomists. “Intermediates” and “adults” were subsequently grouped to ensure consistency between Cycles. Juvenile taxa were reported for all samples and stations separately from adult taxa, as per recommendations in the technical guidance (Environment Canada 2005). Nematodes and Harpacatcoid Copepods were excluded from the invertebrate count data given these organisms are considered meiofauna and typically only a fraction can be retained on a 1 mm sieve (Environment Canada 2005).
Where subsamples (i.e., ½ or ¼ splits) were used instead of whole samples, whole sample abundances were estimated by multiplying subsample abundances by two or four, respectively, and adding any additional larger, fragile organisms removed in the field prior to splitting the samples in the laboratory.
6.2.7.2 Benthic Community Metrics
Four community metrics were calculated to describe benthic invertebrate community composition at each station near Crofton: density, taxa richness, evenness, Bray-Curtis index, and Simpson’s diversity index. Descriptions of each metric are presented below and are further described in the technical guidance document (Environment Canada 2005).
Density
All count data were multiplied by the following density factor (DF) in order to estimate the number of organisms in 1 m2, as required for electronic reporting for the national database (Appendix A7):
DF = 1 m2 / (grab sample area – area of subsamples taken for sed chem)
DF = 1 m2 / [0.1m2 (grab sample area) – 0.0016m2 (redox subsample) – 0.0016m2 (TOC/TN subsample)]
DF = 10.33
Individual densities were reported per replicate, and station densities were calculated using the average of all replicates.
Richness
Taxonomic richness was reported as the “number of different taxa” by counting the number of families (or higher levels, if family not available). Richness was calculated for each station replicate by counting the number of taxa present in each sample. Total station richness was calculated by counting the number of taxa present in all samples combined.
Crofton EEM Cycle Five 6-9 Hatfield Evenness Index
Evenness was quantified for each station following Smith and Wilson (1996). The index takes into consideration the abundance of each taxon in proportion to total abundance, and the taxonomic richness at the station. Evenness was calculated as follows:
s 2 = ∑ []pi1/E /S =1i
where: E = evenness;
th pi = proportion of i taxon at the station; and S = number of taxa in the sample.
A maximum value of “1” indicates entirely equal abundances of each taxon, and a minimum value of “0” indicates entirely unequal abundances.
Simpson’s Diversity Index
Simpson’s diversity index (D) takes into account both the abundance patterns and taxonomic richness of the community. This is calculated by determining, for each taxonomic group at a site, the proportion of individuals that it contributes to the total at the site. This diversity index can range from 0 to 1, with a value of 1 representing the highest diversity. Simpson’s diversity index was calculated as follows:
s 2 −= [] D ∑ p1 i i=1
where: D = Simpson’s diversity index;
th pi = the proportion of the i taxon at the station; and S = number of taxa (family) at the station. Bray-Curtis Index
The Bray-Curtis dissimilarity co-efficient is a distance measurement that reaches a maximum value of “1” for two sites that are entirely different and a minimum of “0” for two sites that possess identical descriptors (Bray and Curtis 1957). Bray-Curtis dissimilarity coefficients were calculated to compare the degree of similarity between individual stations and the reference median. The seven stations, located at the farthest ends of the gradients and exhibiting C:N ratios ranging from approximately 5 to 10, were used to calculate reference median: CRB5A, CRB5B, CRB5C, CRB6, CRB6A, CRB6B and CRB7. Dissimilarity coefficients for the reference median and individual stations were calculated using SYSTAT 10 (SPSS 2000).
Crofton EEM Cycle Five 6-10 Hatfield The Bray-Curtis index is calculated as follows:
n ∑ y − y =1i i1 i2
CB =− n ∑ ()y + y =1i i1 i2
where: B - C = Bray-Curtis distance between sites 1 and 2;
yi1 = count for species I at site 1;
yi2 = count for species I at site 2; and n = total number of species present at the two sites.
6.2.7.3 Statistical Analyses
Statistical analyses were conducted using Excel 2003 and SYSTAT 11 (SPSS 2000).
Summary Statistics
Summary statistics (i.e., mean, median, standard deviations, standard error, and minimum and maximum values) were calculated using Excel for each benthic community metric, as well as supporting sediment and water quality variables, by station.
Correlations
Spearman’s rank correlations were generated in SYSTAT and used to evaluate relationships among benthic invertebrate community metrics, among supporting environmental variables, and between metrics and environmental variables. Correlations with correlation coefficients (rs) greater than the critical rs (two-tailed, α = 0.10) were indicative of statistically significant relationships. Moderate correlations were defined as those ranging from |0.50| to |0.75|. Strong correlations were defined as those ranging from |0.75| to |1.00|.
Regression Analyses
Regression analyses generated in SYSTAT were used to determine if a gradient of effects on benthic invertebrates was evident with the exposure gradient, whether these effects exceed critical levels (see next section), and whether a corresponding gradient of organic matter sources could be identified. C/N ratio was also used as an additional surrogate indicator of pulpmill effluent exposure, rather than distance from the outfalls, which has proved to be a poor indicator of exposure in past Cycles. Regression analyses were used to determine if there was a significant linear relationship between distance from the mill (or C/N ratio as a surrogate) and benthic community metrics, sediment quality variables, and water quality variables.
Crofton EEM Cycle Five 6-11 Hatfield All tests were conducted at a significance level of α = 0.10 (Power = 0.90). Therefore, a significant effect was considered to be a relationship with p < 0.10. Residual plots were evaluated to ensure that following assumptions of the regression model had been met:
Equal variances: residual plots were examined to assess the evenness (homoscedasticity) of distributions of residual error estimates versus the values predicted by the model; Normal distribution of data: residual plots were evaluated for normality, and the Studentized residual generated by SYSTAT was used to evaluate for the presence of outliers; and Independent observations: the Durbin-Watson statistic generated by SYSTAT as a measure of autocorrelation was used to determine whether or not observations were independent from one another.
If data met regression assumptions, analyses were conducted using log10-transformed variables to see if the fit of the model could be improved. If the fit was improved, results for log10-transformed variables were reported. If fit was not improved, results for untransformed variables were reported.
If data failed to meet regression assumptions, analyses were conducted using log10-transformed variables. If assumptions of the model were not met using the transformed variables, regressions were conducted using ranked data.
Power Analyses
Post-hoc power analysis was conducted to verify the ability of regression analyses to detect the critical effect size, which for the purposes of Cycle Five EEM is defined as a relationship between a benthic invertebrate community metric and distance from the pulpmill diffuser with a correlation coefficient (r) of at least |0.707| (Environment Canada 2005, Cohen 1988). This “effect” is equivalent to ±2 times the standard deviation of the reference mean.
Statistical power is a function of sample size (n), variability and magnitude of difference (i.e., effect size) one wishes to detect. Regression analyses were considered to have sufficient power when P≥ 0.90 (α=0.1).
Power analysis was conducted using GPOWER (Faul and Erdfelder 1992).
Critical Effect Size
Results from regression analyses were used to determine whether critical effects on benthic invertebrates were observable near the Crofton mill. Where a statistically significant effect according to regression analysis was one with p < 0.10, an effect exceeding the critical effect size (CES) is defined as a relationship between a benthic invertebrate community metric and distance (or distance surrogate) from the pulpmill diffuser with a correlation coefficient (r) of at least |0.707| (Environment Canada 2005, Cohen 1988). This CES is equivalent to ±2 times the standard deviation of the reference mean.
Crofton EEM Cycle Five 6-12 Hatfield Cluster Analysis
Cluster analysis is a multivariate procedure which can be used to detect natural groupings in benthic invertebrate data. The procedure is based on the relative densities of taxa at each station; taxa that are more abundant tend to influence the cluster analysis more than rare taxa. Cluster analysis was conducted by performing pairwise comparisons between stations using Bray-Curtis dissimilarity coefficients created from density data for individual taxa. Cluster analysis was conducted using hierarchical clustering with average linkages in SYSTAT.
6.2.7.4 Temporal Trends
Benthic invertebrate metrics and statistical results from Cycle Five were compared with historical results in order to evaluate changes that may have occurred in the benthic community or habitat quality near the mill over time. Major changes in these results were discussed in relation to changes in effluent exposure and/or habitat characteristics, if any, between cycles. The focus was on comparisons with Cycles Two and Three, given these cycles used the same gradient design and sampling methods were used in Cycle Five. Comparisons with Cycle One were also be made on a more general qualitative level; Cycle One sampling location, sieve size (0.18 mm), sample splitting protocols, and taxonomic ID level preclude direct comparisons with subsequent cycles.
6.3 RESULTS
6.3.1 Benthic Invertebrate Communities
Table 6.2 summarizes density, taxonomic richness, Simpson’s diversity, and evenness for benthic invertebrate communities sampled in March 2009 near Crofton. Data for adults and juveniles are reported separately in the table; statistical analyses and comparisons were conducted using adult data only.
6.3.1.1 Density and Community Composition
Mean adult density of benthos ranged from 3,621 adults/m2 at CRB7 (Osborn Bay, 3km from outfall) to 9,194 adults/m2 at CRB3 (1.8km south of outfall) (Figure 6.2). Mean density variability was consistent across most stations, with the greatest variability at station CRB5A. Mean densities at near-field stations ranged from 7,716 to 9,058 adults/m2. With the exception of CRB3 and CRB4A, the two gradient stations closer to the mill, mean density values were lower at gradient stations relative to near-field stations, ranging from 3,621 to 6,851 adults/m2.
Crofton EEM Cycle Five 6-13 Hatfield Figure 6.2 Mean adult benthic invertebrate density (± SE) per station, Crofton EEM Cycle Five, March 2009.
Table 6.3 presents the 40 most populous taxa (family level) identified, in decreasing order of adult density. Although over 100 different taxa were identified in Crofton benthic samples, these top 40 taxa account for 97% of the total density of adult organisms collected. The ten most abundant taxa accounted for 85% of total observed density and polychaetes were the dominant taxa observed at all stations.
Twenty-four of the forty most abundant taxa collected in March 2009 were polychaetes. The majority of polychaetes collected belong to the subclass Sedentaria, which includes polychaetes that are generally considered to be non-motile, such as obligate tube-dwellers. A lesser proportion of polychaetes collected belonged to the subclass Errantia (i.e., typically motile polychaetes which may be herbivores, predators, or detritivores).
The most abundant Sedentary polychaetes belonged to the family Capitellidae. Capitellids are small, errant, burrowing worms, considered to be opportunistic species in a wide variety of soft bottom environments (Pearson and Rosenberg 1978). These polychaetes were found at all stations, with the highest densities being observed at locations nearest the outfall and the lowest at CRB7, 3km to the south. The second most abundant Sedentary polychaetes belonged to the family Oweniidae. Oweniids are filter and deposit feeders characterized by long tubes composed of overlapping sand grains and shell fragments. These organisms were found at all stations, with the exception of CRB5C, 7.8km to the northwest. The Oweniids were distributed fairly evenly throughout the sample area with the highest concentrations observed at CRB3 and CRB5A (within 2km of the outfalls). The third most abundant taxa were Bivalves (non-polychaetes) from the Lucinidae family. Lucinid densities were similar across stations, but were most abundant at stations CRB4A and CRB7 (Table 6.3).
Crofton EEM Cycle Five 6-14 Hatfield Table 6.2 Summary of benthic invertebrate community statistics, Crofton EEM Cycle Five, March 2009.
Near-Field Stations Northwest Gradient Stations South Gradient Stations Taxa CRB2A CRB2 CRB1A CRB5A CRB4A CRB5B CRB5C CRB3 CRB7 CRB6A CRB6B CRB6 Exposure Gradient Distance from diffuser (km) 0.60 0.10 0.10 1.90 3.30 5.00 7.80 1.80 3.00 3.30 3.80 6.70 C:N Ratio 18.5 17.7 17.5 6.7 16.3 6.3 8.6 10.6 7.5 7.1 5.0 10.0 Adult Organisms1 Density Sample 1 10,139 10,038 8,746 11,095 7,038 4,851 5,197 11,980 3,346 5,665 3,437 2,075 Sample 2 7,170 6,194 7,455 4,404 8,563 6,082 2,665 8,726 4,048 4,668 5,787 6,102 Sample 3 5,838 10,943 10,038 5,054 11,116 4,668 3,305 6,875 3,468 4,526 3,854 3,529 Mean Density (n/m2) 7,716 9,058 8,746 6,851 8,906 5,200 3,722 9,194 3,621 4,953 4,360 3,902 Median Density 7,170 10,038 8,746 5,054 8,563 4,851 3,305 8,726 3,468 4,668 3,854 3,529 SD of Density 2,202 2,522 1,292 3,690 2,061 769 1,317 2,585 375 621 1,253 2,039 SE of Density 1,271 1,456 746 2,130 1,190 444 760 1,492 216 358 724 1,177 Taxonomic Richness Sample 1 363634452848154727363543 Sample 2 343938433043144535403738 Sample 3 353939402340183529473944 Mean Taxonomic Richness 35 38 37 43 27 44 16 42 30 41 37 42 Median for Richness 35 39 38 43 28 43 15 45 29 40 37 43 SD of Taxa Richness 123344264623 SE of Taxa Richness 112122142312 Total Station Taxa Richness 55 49 49 57 38 61 23 60 42 64 55 63 Evenness Index 0.061 0.059 0.072 0.068 0.132 0.096 0.166 0.073 0.078 0.125 0.126 0.155 Bray-Curtis Index 2 0.545 0.505 0.543 0.374 0.679 0.201 0.394 0.417 0.412 0.201 0.272 0.310 Simpson's Diversity Index 0.701 0.652 0.717 0.743 0.800 0.829 0.738 0.772 0.696 0.875 0.855 0.897 Juvenile Organisms 2 Mean density (adults/m ) 1047.5 901.7 1098.4 800.0 983.1 932.3 857.7 881.4 1200.1 742.4 1047.5 945.8 Total taxa richness 31 27 33 28 33 32 41 27 22 30 27 45 1 Nematoda and calanoid copepods not included in analyses. 2 Bray-Curtis indices are based on the comparison of individual stations to the "reference median" calculated using stations CRB5A, CRB5B, CRB5C, CRB6, CRB6A, CRB6B, and CRB7.
Crofton EEM Cycle Five 6-15 Hatfield Table 6.3 Forty most abundant benthic taxa, Crofton EEM Cycle Five, March 2009.1
Near-field area Northwest gradient stations South gradient stations Total Major Taxonomic CRB1A CRB2 CRB2A CRB4A CRB5A CRB5B CRB5C CRB3 CRB6 CRB6A CRB6B CRB7 Family Taxa Gr oup N of Crof ton Indian Willy North Escape Alarm S of Booth Parminter Osborn Doc k Pt. All Stations outfall outfall Reef Island Reef Reef Rock outfall Bay Pt. Bay Capitellidae Polychaeta (Sedentaria) 4335.81 5179.92 4145.97 1515.33 352.56 979.71 857.67 1556.01 1084.8 603.42 1101.75 437.31 22150.26 Ow eniidae Polychaeta (Sedentaria) 583.08 1037.34 447.48 27.12 2335.71 1772.97 0 3152.7 891.57 1108.53 332.22 298.32 11987.04 Lucinidae Mollusca: Bivalvia 620.37 457.65 576.3 3467.97 393.24 688.17 972.93 505.11 200.01 922.08 511.89 1939.08 11254.8 Spionidae Polychaeta (Sedentaria) 1393.29 715.29 1698.39 2240.79 54.24 359.34 23.73 264.42 311.88 125.43 603.42 105.09 7895.31 Lumbrineridae Polychaeta (Errantia) 88.14 193.23 298.32 349.17 142.38 132.21 6.78 328.83 484.77 271.2 166.11 135.6 2596.74 Tellinidae Mollusca: Bivalvia 142.38 111.87 37.29 1122.09 61.02 159.33 308.49 67.8 23.73 77.97 71.19 128.82 2311.98 Philomedidae Arthropoda Ostracoda 277.98 166.11 145.77 67.8 108.48 81.36 0 318.66 308.49 67.8 149.16 81.36 1772.97 Thyasiridae Mollusca: Bivalvia 3.39 91.53 88.14 0 47.46 206.79 322.05 64.41 132.21 61.02 54.24 359.34 1430.58 Tubulanidae Nemertea 98.31 108.48 105.09 13.56 61.02 44.07 0 122.04 50.85 108.48 27.12 30.51 769.53 Pilargidae Polychaeta (Errantia) 50.85 71.19 40.68 413.58 20.34 23.73 27.12 50.85 10.17 27.12 3.39 13.56 752.58 Maldanidae Polychaeta (Sedentaria) 47.46 37.29 0 0 108.48 50.85 0 44.07 294.93 40.68 40.68 6.78 671.22 Paraonidae Polychaeta (Sedentaria) 200.01 105.09 135.6 0 16.95 23.73 0 57.63 6.78 27.12 13.56 0 586.47 Lucinoma Mollusca: Bivalvia 50.85 23.73 50.85 10.17 27.12 101.7 6.78 67.8 3.39 71.19 13.56 145.77 572.91 Glyceridae Polychaeta (Errantia) 40.68 57.63 77.97 13.56 30.51 33.9 30.51 61.02 67.8 47.46 30.51 57.63 549.18 Orbiniidae Polychaeta (Sedentaria) 71.19 37.29 54.24 88.14 23.73 16.95 0 0 10.17 16.95 71.19 149.16 539.01 Pholoidae Polychaeta (Errantia) 6.78 20.34 3.39 0 152.55 16.95 0 37.29 240.69 40.68 3.39 10.17 532.23 Phoxocephalidae Arthropoda: Amphipoda 20.34 27.12 6.78 3.39 30.51 20.34 0 125.43 64.41 145.77 37.29 10.17 491.55 Syllidae Polychaeta (Errantia) 3.39 6.78 0 13.56 47.46 67.8 3.39 30.51 108.48 47.46 44.07 10.17 383.07 Terebellidae Polychaeta (Sedentaria) 10.17 6.78 16.95 3.39 50.85 40.68 3.39 20.34 40.68 111.87 54.24 23.73 383.07 Cirratulidae Polychaeta (Sedentaria) 13.56 30.51 40.68 0 27.12 54.24 0 64.41 57.63 30.51 40.68 16.95 376.29 Nephtyidae Polychaeta (Errantia) 54.24 30.51 33.9 122.04 6.78 20.34 27.12 10.17 3.39 16.95 23.73 16.95 366.12 Pectinariidae Polychaeta (Sedentaria) 47.46 61.02 3.39 84.75 40.68 20.34 3.39 54.24 6.78 30.51 3.39 10.17 366.12 Hesionidae Polychaeta (Errantia) 44.07 47.46 3.39 183.06 0 3.39 6.78 6.78 16.95 3.39 0 0 315.27 Lineidae Nemertea 3.39 6.78 37.29 54.24 30.51 16.95 3.39 10.17 91.53 20.34 20.34 6.78 301.71 Oedicerotidae Arthropoda: Amphipoda 20.34 37.29 3.39 13.56 23.73 6.78 0 16.95 40.68 98.31 37.29 0 298.32 Nereididae Polychaeta (Errantia) 61.02 54.24 33.9 3.39 37.29 13.56 0 30.51 20.34 20.34 0 20.34 294.93 Goniadidae Polychaeta (Errantia) 20.34 20.34 33.9 67.8 16.95 20.34 6.78 6.78 27.12 20.34 20.34 13.56 274.59 Columbellidae Mollusca: Gastropoda 30.51 91.53 6.78 47.46 33.9 13.56 0 13.56 3.39 0 16.95 0 257.64 Phyllodocidae Polychaeta (Errantia) 10.17 23.73 6.78 115.26 16.95 3.39 0 33.9 13.56 20.34 3.39 6.78 254.25 Onuphidae Polychaeta (Sedentaria) 13.56 0 6.78 0 47.46 30.51 0 27.12 47.46 16.95 33.9 16.95 240.69 Magelonidae Polychaeta (Sedentaria) 0 3.39 0 3.39 37.29 40.68 0 16.95 94.92 10.17 16.95 3.39 227.13 Scalibregmatidae Polychaeta (Sedentaria) 0 0 6.78 16.95 20.34 23.73 0 13.56 47.46 20.34 0 71.19 220.35 Cossuridae Polychaeta (Sedentaria) 128.82 64.41 20.34 0 0 00003.3900216.96 Aoridae Arthropoda: Amphipoda 77.97 0 0 0 13.56 0 0 30.51 0 0 10.17 0 132.21 LeuconidaeArthropoda: Cumacea3.3900006.78071.1916.953.3927.120128.82 Rutidermatidae Arthropoda: Ostracoda000020.343.390091.533.3910.170128.82 Golfingiidae Spiuncula 6.78 3.39 3.39 3.39 67.8 3.39 0 3.39 13.56 6.78 6.78 0 118.65 Polynoidae Polychaeta (Errantia) 27.12 6.78 0 0 3.39 16.95 3.39 13.56 10.17 23.73 3.39 0 108.48 Amphiuridae Echindermata: Ophiuroide 0 0 3.39 23.73 3.39 0 6.78 0 6.78 54.24 3.39 3.39 105.09 Ampeliscidae Arthropoda: Amphipoda 3.39 13.56 3.39 0 23.73 16.95 0 10.17 6.78 13.56 6.78 3.39 101.7 1 These forty taxa are reduced to family level and account for 97% of total abundance among all stations.
Crofton EEM Cycle Five - Final 6-16 Hatfield 6.3.1.2 Taxonomic Richness
Total numbers of taxa collected at individual stations exhibited variability across stations. The lowest total richness (23 taxa) was observed at the furthest station from the mill CRB5C (Alarm Rock, 7.8km to the northwest) and the highest richness (63 taxa) was at CRB6 (Booth Bay, 6.7km south of the outfall) (Table 6.2, Figure 6.3). Overall stations nearest the Crofton outfalls supported similar numbers of taxa as stations farther along the exposure gradient.
Figure 6.3 Taxonomic richness (total per station) of benthic invertebrate communities, Crofton EEM Cycle Five, March 2009.
70 60 50 40 30 20 10 Total Taxa Richness Taxa Total 0 CRB2 CRB3 CRB6 CRB7 CRB5C CRB5B CRB6B CRB2A CRB1A CRB4A CRB6A CRB5A Near-field Far-f ield Decreasing Effluent Exposure (C:N Ratio)
6.3.1.3 Biotic Community Indices Evenness Index
Evenness ranged from 0.059 at CRB2 (Outfall) to 0.166 at CRB5C (Alarm Rock), the farthest station from the outfalls (Table 6.2, Figure 6.4). Gradient stations exhibited variable evenness; however, evenness at near-field stations was similar and within the lower range of values observed at stations farther from the mill.
Figure 6.4 Evenness of benthic invertebrate communities, Crofton EEM Cycle Five, March 2009.
0.20
0.15
0.10 Evenness 0.05
0.00 CRB2 CRB3 CRB6 CRB7 CRB5B CRB6B CRB5C CRB2A CRB1A CRB4A CRB6A CRB5A Near-field Far-field Decreasing Effluent Exposure (C:N Ratio)
Crofton EEM Cycle Five 6-17 Hatfield Simpson’s Diversity
Simpson’s Diversity within benthic communities was variable across stations (Figure 6.5). Stations closest to the outfalls (CRB1A, CRB2, and CRB2A) ranged from 0.652 to 0.717, while diversity at more distant stations ranged from 0.696 to 0.897 (Table 6.2).
Figure 6.5 Simpson's Diversity across benthic invertebrate communities, Crofton EEM Cycle Five, March 2009.
Bray-Curtis Index
Bray-Curtis indices from the seven stations (i.e., CRB5A, CRB5B, CRB5C, CRB6, CRB6A, CRB6B and CRB7) used to calculate the reference median with exposure stations ranged between 0.201 and 0.412 (Table 6.2, Figure 6.6). Values closer to 0 indicate a more similar community composition to the reference median community.
Stations closest to the outfalls (CRB1A, CRB2, CRB2A and CBR3) exhibited Bray-Curtis values of 0.417 to 0.545, indicating that communities at these stations were only slightly dissimilar to the stations used to establish the reference median. The benthic community at station CRB4A (Willy Island) was least similar to the reference median community with a value of 0.679 (Figure 6.6).
Crofton EEM Cycle Five 6-18 Hatfield Figure 6.6 Bray-curtis similarity indices for benthic communities, Crofton EEM Cycle Five, March 2009.
6.3.1.4 Statistical Analyses Regression Analyses: Community Metrics vs. Exposure Gradient
Regression results between benthic endpoints versus C:N ratio and distance from the outfalls are summarized in Table 6.4. A statistically significant relationship was observed between total density of benthic invertebrates and both C:N ratio (p=0.007) and distance from outfalls (p=0.027). Significant relationships were also found between the Bray-Curtis index and C:N ratio (p=0.001), Simpson’s diversity and C:N ratio (p=0.067), evenness and distance (p=0.000), Bray-Curtis index and distance (p=0.035), and Simpson’s diversity and distance (p=0.015).
Total adult density and Bray-Curtis index were greater with increased effluent exposure, as represented by either a higher C:N ratio or decreased distance from the outfalls. In contrast, evenness increased with distance and Simpson’s diversity decreased with C:N ratio and increased with distance, indicating that evenness and diversity increase with decreased effluent exposure. Plots of these significant relationships are presented in Figure 6.7. Taxonomic richness was not significantly correlated (p ≥ 0.10) with C:N ratio or distance from the outfalls, and evenness was not significant when correlated with C:N ratio.
Critical Effects Size Analyses
Results of regression analyses performed to test relationships between benthic community variables and the exposure gradient were evaluated against the critical effects size (CES) criteria (r > |0.707|) in Table 6.4. When distance from the outfalls was used as the exposure gradient near Crofton, the only benthic community variable that exceeded the CES was evenness. When C:N ratio was used as a surrogate for the exposure gradient, density and Bray-Curtis index both showed differences among stations that exceeded the CES.
Crofton EEM Cycle Five 6-19 Hatfield Table 6.4 Results of regressions and correlations conducted to test relationships between benthic community variables and gradients of pulpmill effluent exposure, Crofton EEM Cycle Five, March 2009.
A. Using C:N ratio as an indicator of effluent exposure (i.e., independent variable) Dependent p -value for Correlation coefficients Direction of Critical 2 1 2 Variable F-test Regression Equation r r rs Effect? Effect Effect? Ranked Total Density (adult) 0.007 Rank Total Density = 1.773 + 0.727*Ranked C:N 0.727 0.529 0.727 Yes increase with C:N ratio Yes Ranked Total Richness (adult) 0.252 Ranked Total Richness = 8.697 - 0.364*Ranked C:N 0.359 0.129 -0.337 No - No Ranked Evenness 0.183 Ranked Evenness = 9.182 - 0.413*Ranked C:N 0.413 0.170 -0.413 No - No Bray-Curtis 0.001 Bray-Curtis = 0.138 + 0.024*C:N 0.841 0.707 0.855 Yes increase with C:N ratio Yes Ranked Simpson's Diversity 0.067 Ranked Diversity = 10.045 - 0.545*Ranked Diversity 0.545 0.298 -0.545 Yes decrease with C:N ratio No
B. Using distance from outfalls as an indicator of effluent exposure (i.e., independent variable) Dependent p -value for Correlation coefficients 2 Variable F-test Regression Equation r r rs Effect? Ranked Total Density (adult) 0.027 Ranked Total Density = 10.409 - 0.617*Ranked Distance 0.634 0.401 -0.614 Yes decrease with distance No Ranked Total Richness (adult) 0.621 Ranked Total Richness =5.338 + 0.157*Ranked Distance 0.159 0.025 0.116 No - No Evenness 0.000 Evenness = 0.058 + 0.014 Distance 0.884 0.782 0.902 Yes increase with distance Yes Ranked Bray-Curtis 0.035 Ranked Bray-Curtis = 10.312 - 0.615*Ranked Distance 0.611 0.373 -0.613 Yes decrease with distance No
Log10 Simpson's Diversity 0.015 Log10 Diversity = 0.829+0.046*Log10 Diversity 0.682 0.465 0.660 Yes increase with distance No Bolded entries represent statistically significant relationships (α = 0.10). r = Pearson's correlation coefficient (parametric correlations). r2 = coefficient of determination. rs = Spearman's rank correlation coefficient (non-parametric correlations). 1 p < 0.10 2 critically significant regression effect r > |0.707|
Crofton EEM Cycle Five 6-20 Hatfield Figure 6.7 Significant regressions of mean adult density, Bray-Curtis Index, and evenness against C:N ratio and distance from outfalls, showing 95% confidence intervals, Crofton EEM Cycle Five, March 2009.
Power Analysis of Regressions The power of regression analyses to detect an “effect” (r = |0.707|) on benthic communities using n=12 stations along the effluent exposure gradient was 0.94 (Appendix A8); this was above the recommended level of P = 0.90, and sufficient power was therefore available to detect an effect. Correlations Among Community Metrics The results of Spearman rank correlations among benthic invertebrate community metrics are presented in Table 6.5. Moderately significant inverse correlations were observed between: density and evenness, richness and Bray-Curtis index, and Bray-Curtis index and Simpson’s diversity. Moderately significant positive correlations were present between: density and Bray-Curtis index, and evenness and Simpson’s diversity. Significant correlations are represented graphically in Figure 6.8.
Crofton EEM Cycle Five 6-21 Hatfield Table 6.5 Results of Spearman rank correlations (rs) among all benthic invertebrate metrics (n=12), Crofton EEM Cycle Five, 2009.
Density (n/m2) Richness Evenness Bray-Curtis Index Density (n/m2) Richness 0.242 Evenness -0.573 -0.284 Bray-Curtis Index 0.564 -0.537 -0.375 Simpson's Diversity -0.231 0.392 0.657 -0.637
Figure 6.8 Significant Spearman rank correlations of benthic metrics, Crofton EEM Cycle Five, 2009.
14 Evenness and Density 14 Bray-Curtis Index and Density 12 12 10 10 8 8 6 6 4 4 Ranked Bray-Curtis Ranked Evenness 2 2 rs=0.564 rs=-0.573 0 0 0510150 5 10 15 Rank e d De ns ity Ranked Density
14 Bray-Curtis Index and Richness Simpson's Diversity and Evenness 14 12 12 10 rs=-0.537 10 r =0.657 8 s 8 6 6 4 4 Diversity 2
Ranked Bray-Curtis Bray-Curtis Ranked 2 0 Ranked Simpson's 0 0 5 10 15 0 5 10 15 Ranked Richness Ranked Evenness
Sim pson's Dive rsity a nd Bra y-Curtis 14 12 10 8
Diversity 6 4 rs=-0.637 Ranked Simpson's 2 0 0 5 10 15 Ranked Bray-Curtis Index
Crofton EEM Cycle Five 6-22 Hatfield Cluster Analysis Among Communities
Results of cluster analysis using Bray-Curtis dissimilarity indices are presented visually in Figure 6.9. Station CRB4A (Willy Island) was most dissimilar from all other stations; stations CRB5C (Alarm Rock) and CRB7 (Osborn Bay) were also somewhat dissimilar from other stations. The three stations nearest the Crofton outfalls (i.e., CRB1A, CRB2 and CRB2A) clustered together, indicating similar community composition at these stations. Another cluster included CRB3, CRB5A, CRB5B and CRB6A; this group was relatively similar to stations CRB6 and CRB6B. Statistical distances between individual stations ranged from approximately 0.10 to 0.65.
Figure 6.9 Dendrogram describing similarity of benthic invertebrate communities at station sampled for Crofton EEM Cycle Five, based on Bray-Curtis dissimilarity coefficients among stations.
Crofton EEM Cycle Five 6-23 Hatfield 6.3.2 QA/QC and Verifications
As per the Technical Guidance Document, the following QA/QC procedures were used during laboratory analysis:
Four samples (1A-3, 2A-1, 2A-2 and 2A-3) were split to test if splitting was an option for larger samples (Appendix A7);
Splitting was confirmed as a reliable sampling option for larger samples;
Only two samples required splitting (4A-1 at ½ and 4A-3 at ¼). Although other samples were in multiple jars (3 – 11) the substrate was coarse gravel and these samples were treated in their entirety;
All of the samples were subjected to QA at 20%, 50%, or 100% (Appendix A7). Sorting error did not exceed 4% in any samples; therefore, none required re-sorting;
Appropriate taxonomic references were used according to study area location and habitat; and
Over the course of the EEM program, a reference collection of taxa identified in EEM benthic samples was created; however, the reference collection from Cycle Two was inadvertently discarded and a new collection was started using Cycle Three samples. In Cycle Five, new taxa that were not identified during previous cycles were verified by Biologica Environmental Services (Appendix A7) before inclusion in the reference collection. These invertebrates were added to the cumulative collection and the master list amended accordingly. The collection was returned to Hatfield for storage and future reference.
6.3.3 Supporting Environmental Variables 6.3.3.1 Habitat Characteristics
Station depth and particle size were measured to assess habitat characteristics at benthic survey stations in support of the invertebrate community evaluation in Stuart Channel (Table 6.6). Samples were collected from the channel at depths ranging between 49.7 and 68 m in the near-field area, and between 44.3 and 61 m at gradient stations. Particle size distribution of sediments collected at each survey station along the gradient are presented graphically in Figure 6.10. Sand was the dominant size fraction at most stations, exceeding 70% at all stations except CRB5C (Alarm Rock) where silt was the most significant fraction at 74% (Table 6.6). With the exception of CRB5C, the fine fraction (silt and clay combined) was greatest at stations nearer to the outfalls. No gravel was present at near-field stations, and this coarser size fraction was only present in minor amounts at some far-field stations (1 – 14%).
Crofton EEM Cycle Five 6-24 Hatfield Figure 6.10 Particle size1 distribution of sediments at benthic invertebrate community survey stations, Crofton EEM Cycle Five, 2009.
Gravel (%) Sand (%) Silt (%) Clay (%) 100%
90%
) 80%
70%
60%
50%
40%
30%
Fraction of Sediment Sample (% 20%
10%
0% CRB2 CRB3 CRB6 CRB7 CRB5B CRB5C CRB6B CRB1A CRB2A CRB4A CRB5A CRB6A Near-field Northw est stns South stns 1 Gravel = >2mm; sand = 0.063 to 2 mm; silt = 0.004 to 0.063 mm; clay = <0.004 mm.
6.3.3.2 Water Quality
Dissolved oxygen (DO), temperature, and salinity were used to evaluate near-bottom water quality in the receiving environment at benthic invertebrate community survey stations (Table 6.6). DO ranged between 7.35 and 9.18 mg/L; water temperature ranged between 7.4 and 7.8 °C; and salinity ranged between 29.2 and 30 ‰.
Water quality variables generally fluctuated between gradient stations, and did not exhibit obvious trends with distance from the mill (Figure 6.11). DO averaged 8.75 mg/L at near-field stations, 8.45 mg/L at far-field northwest stations, and 8.31 mg/L at far-field south stations. Temperature averaged 7.50 °C at near-field stations, 7.53 °C at northwest stations, and 7.56 °C at south stations. Salinity averaged 29.9 ppt at near-field stations, 29.5 ppt at northwest stations, and 29.6 ppt at south stations.
Crofton EEM Cycle Five 6-25 Hatfield Figure 6.11 Near-bottom water quality at benthic invertebrate community survey stations, Crofton EEM Cycle Five, 2009.
9.5 Dissolved Oxygen
9.0
8.5
8.0
7.5 Dissolved Oxygen (mg/L) Oxygen Dissolved
7.0 CRB2 CRB3 CRB6 CRB7 CRB1A CRB2A CRB4A CRB5A CRB5B CRB6A CRB6B CRB5C
Near-field Northwest stns South stns
8.0 Temperature 7.9 7.8 7.7 7.6 7.5 7.4 7.3
Temperature (deg. C) (deg. Temperature 7.2 7.1 7.0 CRB2 CRB3 CRB6 CRB7 CRB1A CRB2A CRB4A CRB5A CRB5B CRB6A CRB6B CRB5C Near-field Northwest stns South stns
30.0 Salinity 29.9 29.8 29.7 29.6 29.5 29.4
Salinity (ppt) 29.3 29.2 29.1 29.0 CRB3 CRB6 CRB7 CRB2 CRB6A CRB6B CRB1A CRB2A CRB4A CRB5A CRB5B CRB5C Near-field Northwest stns South stns
Crofton EEM Cycle Five 6-26 Hatfield Table 6.6 Habitat characteristics, near-bottom water quality, and sediment quality at benthic invertebrate community survey stations, Crofton EEM Cycle Five, 2009.
Habitat Characteristics Water Quality Sediment Quality Station Distance Depth Particle Size2 (%) D.O.1 Temp.1 Salinity TOC1,3 TN1,3 C:N1,3 Redox3,4 Sulphides3 (km) (m) Gravel Sand Silt Clay (mg/L) (C) (ppt) (%) (%) Ratio (mV) (μmol) Near-field stations CRB1A (North of outfall) 0.1 59.7 <1 74 20 5 8.68 7.6 30.0 2.10 0.12 17.5 -13 60 CRB2 (Outfall) 0.1 68.0 <1 76 19 6 8.96 7.5 29.8 2.07 0.12 17.7 14 58 CRB2A (Indian Reef) 0.6 49.7 <1 83 12 5 8.60 7.4 29.8 1.60 0.09 18.5 2 75 Average 0.3 59.1 <1 78 17 5 8.75 7.5 29.9 1.92 0.11 17.9 1 64 Northwest stations CRB5A (North Reef) 1.9 49.6 5 83 9 4 8.80 7.5 29.7 0.47 0.07 6.7 28 77 CRB4A (Willy Island) 3.3 49.7 1 72 21 6 8.59 7.4 29.4 1.63 0.10 16.3 35 49 CRB5B (Escape Reef) 5.0 61.0 <1 91 4 4 7.78 7.8 29.2 0.33 0.05 6.3 112 17 CRB5C (Alarm Rock) 7.8 54.1 <1 10 74 16 8.64 7.4 29.8 3.57 0.41 8.6 45 39 Average 4.5 53.6 <1 64 27 8 8.45 7.5 29.5 1.50 0.16 9.5 55 46 South stations CRB3 (South of outfall) 1.8 50.6 4 73 17 6 8.53 7.5 29.2 1.13 0.11 10.6 112 12 CRB7 (Osborn Bay) 3.0 46.0 <1 90 7 2 8.00 7.6 29.8 0.30 0.04 7.5 156 83 CRB6A (Parminter Pt.) 3.3 49.2 1 89 7 3 8.48 7.5 29.7 0.50 0.07 7.1 28 37 CRB6B (Dock Pt.) 3.8 45.0 <1 92 5 3 9.18 7.5 29.5 0.20 0.04 5.0 73 21 CRB6 (Booth Bay) 6.7 44.3 14 77 7 3 7.35 7.7 29.9 0.67 0.07 10.0 87 62 Average 3.7 47.0 <1 84 9 3 8.31 7.6 29.6 0.56 0.07 8.0 91 43
1 D.O. = dissolved oxygen; Temp. = w ater temperature; TOC = total organic carbon; TN = total nitrogen; C:N = carbon/nitrogen ratio. 2 Gravel = >2mm; sand = 0.063 to 2 mm; silt = 0.004 to 0.063 mm; clay = <0.004 mm. 3 Station means calculated using average of three replicates. 4 Temperature-corrected redox potential value used. n/a = reading not available.
Crofton EEM Cycle Five 6-27 Hatfield 6.3.3.3 Sediment Quality Visual Observations
Visual observations of sediments were made during field sampling; photographs of sediment grabs taken at each station are presented on the CD that accompanies this report. The following key observations qualitatively support actual measured levels of organic content and oxidative state in sediments at each station:
Most stations were characterized by odourless greeny-brown sand/mud, including CRB1A (North of outfall), CRB2 (Outfall), CRB3 (South of outfall), CRB2A (Indian Reef), and CRB4A (Willy Island), CRB5B (Escape Reef), CRB6A (Parminter Pt.), CRB6B (Dock Pt.) and CRB7 (Osborn Bay); CRB5A (North Reef) was characterized by odourless, greeny-brown coloured sand/mud with a small amount of gravel; CRB5C (Alarm Rock) was characterized by very soft, odourless mud with some sand; and CRB6 (Booth Bay) was characterized by odourless, greeny-brown coloured sand, mud and some gravel.
Organic Content
Organic content in sediments was evaluated in support of the benthic invertebrate community survey near Crofton by analyses of three measures: total organic carbon (TOC), total nitrogen (TN), and carbon:nitrogen (C:N) ratio. Results are presented in Table 6.6.
Mean sediment C:N ratios were higher in the near-field (17.9) than at far-field stations (8.8) (Table 6.6). Given that in past EEM Cycles C:N ratios in sediments were significantly correlated with the effluent exposure gradient, for this Cycle C:N ratios were used in lieu of distance as a primary indicator of the effluent exposure gradient in order to evaluate other parameters.
Mean sediment TOC levels in the near-field (1.92 %) were higher than those measured at far-field stations (0.56 %) (Table 6.6). The lowest TOC concentration was found at south far-field station CRB6B (0.20 %); the highest concentration was also identified at a far-field station (CRB5C, 3.57 %), although this anomaly was likely due to the dominance of silt and clay at the site relative to the other more sandy sites in the study area. When CRB5C was excluded from the analysis, mean sediment TOC at far-field stations was much lower (0.65 %). In general, with the exception of station CRB5C, TOC levels appeared to generally decline along the declining C:N exposure gradient (Figure 6.12).
Mean sediment TN levels in the near-field (0.11 %) were the same as those measured at far-field stations (0.11 %). TN was elevated at station CRB5C (0.41 %) due to levels of coarse sand uncharacteristic of all other sites in the study area; when CRB5C was excluded from the analysis, average sediment TN in the
Crofton EEM Cycle Five 6-28 Hatfield far-field (0.07 %) was lower than at near-field stations. In general, with the exception of station CRB5C, TN levels appeared to generally decline along the declining C:N exposure gradient (Figure 6.12).
Relative to Cycle Three (2003), TOC and C:N ratio have declined throughout the study area (Figure 6.12). At the gradient stations, TN has also declined since Cycle Three; however, at near-field stations TN has remain relatively unchanged.
Oxidative State
Redox potential and total sulphides were used to evaluate the oxidative state of sediments in support of the benthic invertebrate community survey near Crofton. Results are presented in (Table 6.6).
Mean redox potential was lower in the near-field (0.8 mV) relative to far-field stations (75 mV) (Table 6.6). Redox potential was negative (-13 mV) at only one station, CRB1A, closest to the outfalls. Values at all other stations were positive. Redox potential was highest at station CRB7, 3 km to the south of the outfalls (156 mV). In general, although redox potential was lower at near-field stations than further down the exposure gradient, redox potential within the far-field fluctuated and did not appear to vary consistently along the decreasing exposure gradient (Figure 6.13).
Mean total sulphides were marginally higher in the near-field (64 μmol) compared to far-field stations (44 μmol). Sulphides were highest at far-field station CRB7, 3 km south of the the outfalls, where levels were recorded at 83 μmol; the lowest value (12.1 μmol) was recorded in the far-field at station CRB3, 1.8 km south of the outfalls. In general, sulphides nearer to the mill appeared to decline along the decreasing effluent exposure gradient, while concentrations further down the gradient fluctuated and did not appear to vary consistently (Figure 6.13).
Relative to Cycle Three (2003), redox potential has increased and sulphides have decreased declined throughout the study area (Figure 6.13).
Crofton EEM Cycle Five 6-29 Hatfield Figure 6.12 TOC, TN, and C:N ratio at benthic invertebrate community survey stations, Crofton EEM Cycle Three (2003) and Cycle Five (2009).
30.0 C:N Ratio
25.0 Cycle Five (2009) Cycle Three (2003) 20.0 o
15.0 C:N Rati C:N 10.0
5.0
0.0 CRB2 CRB3 CRB6 CRB7 CRB2A CRB1A CRB4A CRB6A CRB5A CRB5B CRB6B CRB5C
4.0 Total Organic Carbon 3.5 ) Cycle Five (2009) Cycle Three (2003) 3.0
2.5
2.0
1.5
1.0 Total Organic Carbon (% 0.5
0.0 CRB2 CRB3 CRB6 CRB7 CRB2A CRB1A CRB4A CRB6A CRB5A CRB5B CRB6B CRB5C
0.45 Total Nitrogen 0.40 Cycle Five (2009) Cycle Three (2003)
) 0.35 0.30 0.25 0.20 0.15
Total Nitrogen (% Nitrogen Total 0.10 0.05 0.00 CRB2 CRB3 CRB6 CRB7 CRB2A CRB1A CRB4A CRB6A CRB5A CRB5B CRB6B CRB5C Near-field Far-f ield Cycle 5 Decreasing Effluent Exposure (C:N Ratio)
Crofton EEM Cycle Five 6-30 Hatfield Figure 6.13 Redox potential and sulphides at benthic invertebrate community survey stations, Crofton EEM Cycle Three (2003) and Cycle Five (2009).
200 Redox Potential 150 100 50 0 -50 -100 -150 Redox Potential (mV) -200 Cycle Five (2009) Cycle Three (2003) -250 -300 CRB2 CRB3 CRB6 CRB7 CRB2A CRB1A CRB4A CRB6A CRB5A CRB5B CRB6B CRB5C
450 Sulphides 400 350 Cycle Five (2009) Cycle Three (2003) 300 Mol) μ 250 200 150
Sulphides ( 100 50 0 CRB2 CRB3 CRB6 CRB7 CRB2A CRB1A CRB4A CRB6A CRB5A CRB5B CRB6B CRB5C Near-field Far-f ield Decreasing Effluent Exposure (C:N Ratio)
6.3.3.4 Statistical Analyses Regression Analyses: Supporting Variables vs. Exposure Gradient
Results of regression analyses performed to test relationships between sediment quality, near-bottom water quality, and the exposure gradient (both according to distance from the outfalls and C:N ratio) are summarized in Table 6.7.
Redox potential moderately significantly increased with distance from the outfall (rs = 0.555). None of the other sediment quality variables and none of the near-bottom water quality variables varied significantly when distance from the outfalls was used as the exposure gradient near Crofton.
Crofton EEM Cycle Five 6-31 Hatfield Table 6.7 Results of regression analyses conducted to test relationships between supporting environmental variables and gradients of pulpmill effluent exposure (distance from the outfalls and C:N), Crofton EEM Cycle Five, 2009.
Category p -value Correlation coefficients Strength of Direction of Effect 2 1 2 Dependent Variable for F-test Regression Equation r r rs Ef f e ct ? Effect? EXPOSURE GRADIENT: DISTANCE FROM OUTFALL Sediment Quality Total Organic Carbon 0.367 rank TOC = 8.265 - 0.279*rank distance - - -0.263 No - None Total Nitrogen 0.289 rank TN = 8.539 - 0.322*rank distance ---0.316 No - None Total Sulphides 0.341 sulphides = 58.308 - 2.96*distance -0.301 0.091 -0.298 No - None Redox Potential 0.048 rank redox = 2.927 + 0.564*rank distance --0.555 Yes increase w ith distance Moderate % Gravel 0.637 rank % gravel = 5.659 + 0.133*rank distance - - 0.153 No - None % Sand 0.672 rank % sand = 5.659 + 0.133* rank distance --0.120 No - None % Silt 0.307 rank % silt = 8.476 - 0.312*rank distance ---0.304 No - None % Clay 0.509 rank % clay = 7.782 - 0.202*rank distance ---0.202 No - None Water Quality Dissolved Oxygen 0.103 DO = 8.787 - 0.103*distance -0.493 0.243 -0.407 No - None Temperature 0.766 rank temp = 5.933 + 0.090*rank distance --0.090 No - None Salinity 0.515 rank salinity = 7.761 - 0.199*rank distance - - -0.204 No - None EXPOSURE GRADIENT: C:N RATIO Sediment Quality Total Organic Carbon 0.004 rank TOC = 1.545 + 0.762*rank C:N - - 0.762 Yes increase w ith C:N ratio Strong Total Nitrogen 0.012 rank TN = 2.023 + 0.689*rank C:N - - 0.696 Yes increase w ith C:N ratio Moderate Total Sulphides 0.366 rank sulphides = 4.636 + 0.287*rank C:N - - 0.287 No - None Redox Potential 0.070 rank redox = 10.000 - 0.538*rank C:N - - -0.539 Yes decrease w ith C:N ratio Moderate % Gravel 0.781 rank % gravel = 7.023 - 0.080*rank C:N - - -0.090 No - None % Sand 0.029 rank % sand = 10.568 - 0.626* rank C:N - - -0.627 Yes decrease w ith C:N ratio Moderate % Silt 0.014 rank % silt = 2.091 + 0.678*rank C:N - - 0.683 Yes increase w ith C:N ratio Moderate % Clay 0.059 rank % clay = 2.932 + 0.549*rank C:N - - 0.559 Yes increase w ith C:N ratio Moderate Water Quality Dissolved Oxygen 0.762 rank DO = 5.864 + 0.098*rank C:N --0.098 No - None Temperature 0.268 rank temp = 8.659 - 0.332*rank C:N ---0.348 No - None Salinity 0.304 log salinity = 3.375 + 0.006*log C:N 0.324 0.105 0.454 No - None Bolded entries represent statistically significant relationships (α = 0.10). r = Pearson's correlation coefficient (parametric correlations). r2 = coefficient of determination.
rs = Spearman's correlation coefficient (non-parametric correlations). 1 p < 0.10. 2 Strength evaluated using Spearman rank correlation coefficient: w eak correlation (rs < 0.5), moderate correlation (i.e., 0.5 < rs < 0.75), strong correlation (i.e., rs > 0.75).
Crofton EEM Cycle Five 6-32 Hatfield When C:N ratio was used as a surrogate for the exposure gradient, several sediment quality variables demonstrated significant effects. The following sediment quality variables increased with increasing C:N ratio (i.e., increased effluent exposure): total organic carbon, total nitrogen, % silt, and % clay. Redox potential and sand declined significantly with increasing C:N ratio. With the exception of the strong relationship between TOC and C:N ratio (rs = 0.762), the remaining significant relationships were only of moderate strength (i.e., 0.50 < rs < 0.75). None of the water quality variables demonstrated significant effects using C:N ratio as the exposure gradient.
Correlations Among Supporting Variables
The results of Spearman rank correlations among sediment and near-bottom water quality variables measured near Crofton are presented in Table 6.8.
Relationships among several sediment quality variables were significant: strongly significant inverse correlations were identified between sand versus silt, clay, TOC, and TN. Strongly significant positive correlations were identified between silt versus clay, TOC, and TN, between clay versus TOC and TN, and between TOC versus TN and C:N ratio. A moderately significant positive correlation existed between TN and C:N ratio, and moderately significant inverse correlations were present between redox potential and TOC, TN, and C:N ratio.
No relationships among near-bottom water quality variables were significant. However, a few relationships were identified between sediment quality and near-bottom water quality: moderately significant inverse relationships were present between redox potential (sediments) and DO (water), and silt/clay (sediments) and temperature (water); a moderately significant positive relationship was observed between sulphides (sediments) and salinity (water).
Crofton EEM Cycle Five 6-33 Hatfield Table 6.8 Results of Spearman rank correlations (rs) among sediment and near-bottom water quality variables (n=12), Crofton EEM Cycle Five, 2009.
Sediment Quality Near-bottom Water Quality Gravel Sand Silt Clay TOC TN C:N Redox Sulphides DO Temp Salinity Sediment Quality Gravel Sand -0.211 Silt -0.043 -0.928 Clay -0.111 -0.827 0.839 TOC -0.109 -0.883 0.908 0.826 TN -0.059 -0.896 0.907 0.887 0.968 C:N -0.090 -0.627 0.683 0.559 0.762 0.696 Redox 0.157 0.256 -0.462 -0.323 -0.567 -0.517 -0.539 Sulphides 0.039 0.039 0.099 -0.288 0.021 -0.099 0.287 -0.249 Near-bottom Water Quality DO -0.328 -0.109 0.359 0.349 0.231 0.315 0.098 -0.550 0.028 Temp 0.057 0.469 -0.619 -0.591 -0.450 -0.459 -0.348 0.400 0.029 -0.468 Salinity -0.170 -0.197 0.277 -0.093 0.418 0.292 0.454 -0.437 0.672 0.086 0.125
Crofton EEM Cycle Five 6-34 Hatfield Correlations Between Community Metrics and Supporting Variables
The results of Spearman rank correlations between benthic community metrics and supporting sediment and near-bottom water quality variables measured near Crofton are presented in Table 6.9. Scatterplots showing significant relationships are displayed in Figure 6.14.
Several significant relationships were present between benthic metrics and sediment quality variables: density demonstrated moderately significant positive correlations with clay, TN, and C:N ratio; richness demonstrated a moderately significant inverse correlation with silt; Bray-Curtis indices demonstrated strongly significant positive correlations with silt and C:N ratio, moderately positive correlations with clay, TOC, and TN, and a moderately significant inverse relationship with sand; diversity demonstrated a moderately significant positive relationship with gravel and an inverse relationship with C:N ratio. Evenness did not demonstrate any significant relationships with sediment quality variables.
Only one significant relationship was present between benthic community metrics and near-bottom water quality: richness demonstrated a moderately significant positive relationship with temperature.
Table 6.9 Results of Spearman rank correlations (rs) between benthic community metrics and supporting variables (n=12), Crofton EEM Cycle Five, 2009.
Benthic Community Metrics Density Richness Evenness Bray Curtis Diversity Sediment Quality Gravel 0.137 0.462 0.195 -0.151 0.551 Sand -0.410 0.361 -0.130 -0.618 0.235 Silt 0.423 -0.550 -0.049 0.751 -0.479 Clay 0.591 -0.262 -0.093 0.576 -0.363 TOC 0.413 -0.413 -0.056 0.630 -0.392 TN 0.505 -0.290 -0.127 0.577 -0.399 C:N 0.566 -0.354 -0.413 0.855 -0.545 Redox -0.420 0.179 0.448 -0.416 0.301 Sulphides -0.252 -0.256 -0.322 0.343 -0.476 Near-bottom Water Quality DO 0.301 -0.228 -0.364 0.273 -0.392 Temp -0.242 0.585 -0.022 -0.488 0.234 Salinity -0.286 -0.351 -0.107 0.254 -0.372
Crofton EEM Cycle Five 6-35 Hatfield Figure 6.14 Scatterplots showing significant Spearman rank correlations (rs) and best-fit regression line between benthic community metrics and supporting variables, Crofton EEM Cycle Five, 2009.
14 Density vs. Clay 14 Density vs. TN 14 Density vs. C:N 12 12 12 10 10 10 8 8 8 6 6 6 Rank Density Rank r = 0.591 Density Rank r = 0.505 Density Rank 4 s 4 s 4 rs= 0.566 2 2 2 0 0 0 0 5 10 15 0 5 10 15 051015 Rank Clay (%) Rank TN (%) Rank C:N Ratio
14 Richness vs. Silt 14 Richness vs. Water Temperature 14 Bray-Curtis vs. Sand 12 12 12 10 10 10 rs= -0.550 8 8 8
6 6 rs= 0.585 6 rs= -0.618 Rank Richness Rank Rank Richness
4 4 Rank BrayCurtis 4 2 2 2 0 0 0 0 5 10 15 0 5 10 15 0 5 10 15 Rank Silt (%) Rank Temp (deg. C) Rank Sand (%)
14 Bray-Curtis vs. Silt 14 Bray-Curtis vs. Silt 14 Bray-Curtis vs. Clay 12 12 12 10 10 10 8 8 8 rs= 0.751 rs= 0.751 rs= 0.576 6 6 6 Rank Bray Curtis Rank Bray Curtis 4 4 Rank Bray Curtis 4 2 2 2 0 0 0 0 5 10 15 0 5 10 15 051015 Rank Silt (%) Rank Silt (%) Rank Clay (%)
14 Bray-Curtis vs. TOC 14 Bray-Curtis vs. TN 14 Bray-Curtis vs. C:N 12 12 12 10 10 10 8 8 rs= 0.630 rs= 0.577 8 rs= 0.855 6 6 6 Rank Bray Curtis Rank Bray Curtis 4 4 Rank Bray Curtis 4 2 2 2 0 0 0 0 5 10 15 0 5 10 15 0 5 10 15 Rank TOC (%) Rank TN (%) Rank C:N Ratio
14 Diversity vs. Gravel 14 Diversity vs. C:N 12 12 10 10 8 8 rs= 0.551 rs= -0.545 6 6 Rank Diversity Rank Diversity 4 4 2 2 0 0 051015051015 Rank Gravel (%) Rank C:N Ratio 6.4 DISCUSSION
6.4.1 Effects Along the Exposure Gradient
In order to evaluate the effects of mill effluent discharges during Cycle Five, changes in benthic invertebrate communities were assessed along an exposure gradient, both according to distance from the outfalls and C:N ratio. Regression analyses were used to evaluate whether or not community metrics (i.e., density, richness, diversity, Bray-Curtis, and evenness) changed significantly along the gradient; any significant relationship constituted an “effect”. Any relationship with r > |0.707| was classified, as per the Technical Guidance, as exceeding the critical effect size (CES). A summary of effects observed on benthic invertebrate communities along the exposure gradient, both according to C:N ratio and distance from the outfalls, are presented in Table 6.10 and Table 6.11.
Crofton EEM Cycle Five 6-36 Hatfield Table 6.10 Summary of observed responses of benthic invertebrate communities with C:N ratio exposure gradient, Crofton EEM Cycle Five, March 2009.
Exceeds Effect Endpoint Effect? Direction Magnitude CES? Cycle Three Density Yes No Increase with C:N ratio |r|=0.413 Richness No No - - Evenness No No - - Bray-Curtis No No - - Cycle Five Density Yes Yes Increase with C:N ratio |r|=0.727 Richness No No - - Evenness No No - - Increasingly different from Bray-Curtis Yes Yes |r|=0.855 reference with C:N ratio
Table 6.11 Summary of observed responses of benthic invertebrate communities with distance from outfalls, Crofton EEM Cycle Five, March 2009.
Exceeds Effect Endpoint Effect? Direction Magnitude CES? Cycle Three Density Yes No Decrease with distance |r|=0.664 Richness No No - - Evenness No No - - Bray-Curtis No No - - Cycle Five Density Yes No Decrease with distance |r|=0.614 Richness No No - - Evenness Yes Yes Increase with distance |r|=0.902 Increasingly similar to reference Bray-Curtis Yes No |r|=0.613 with distance from outfalls
During Cycle Five, according to density and Bray-Curtis index, effects were found on benthic invertebrate communities along the effluent exposure gradient near Crofton according to C:N ratio (Table 6.10). Invertebrate densities increased with increasing C:N ratio (i.e., higher terrestrial organic content), while according to Bray-Curtis, exposed benthic communities became increasingly different from the reference community with increased C:N ratio. Both of these effects exceeded the CES.
Crofton EEM Cycle Five 6-37 Hatfield Effects on density and Bray-Curtis were also observed when distance from the outfalls was used as the exposure gradient; in addition, evenness varied significantly with distance (Table 6.11). Density was found to decrease with distance from the outfalls, although this effect did not exceed the CES. Evenness was found to increase with distance, and the effect exceeded the CES. Bray-Curtis demonstrated that exposed benthic communities became increasingly similar to the reference community with distance from the outfalls, although this effect did not exceed the CES.
In summary, benthic communities in Cycle Five near Crofton appeared to be enriched closer to the mill, within sediments characterized by higher levels of terrestrial organic content. Closer to the mill, communities became less similar to the reference community and exhibited more uneven numbers of taxa dominated by Capitellid polychaetes, a known opportunistic species of worm found to thrive in organically impacted, soft sediment environments (Pearson and Rosenberg 1978).
In comparison, during Cycle Three, densities of invertebrates also increased closer to the mill, within sediments characterized by higher levels of terrestrial organic content (Table 6.10; Table 6.11). Significant effects were not found according to community richness, evenness, or Bray-Curtis.
6.4.2 Grades of Impact Along the Exposure Gradient
The technical guidance (Environment Canada 2005) provides the following guidance for further examining critical effects or impacts of sediment chemistry on marine benthic invertebrates, based on studies undertaken in the vicinity of salmon aquaculture farms on the east coast of Canada (Poole et al. 1978, Hargrave et al. 1995, cited in Environment Canada 2005) (Table 6.12). For pulp and paper mills, total organic carbon, total nitrogen, redox potential and total sulphides have been identified as variables that may be related to effluent.
Normal concentrations of TOC, based on a survey of control and impacted sediments in the vicinity of Washington State fishfarms, were defined as 1.7% TOC for sediments composed of 20 to 50% silt/clay, 3.2% TOC for sediments composed of 50 to 80% silt/clay, and 2.6% TOC for sediments composed of 80 to 100% silt/clay. Given that sediments within the Crofton study area ranged between 8 and 27 % silt/clay, with the exception of one far-far-field station where silt/clay measured 90%, the 1.7% TOC value was used as “normal” (Table 6.12).
Crofton EEM Cycle Five 6-38 Hatfield Table 6.12 Scheme used to grade impacts of sediment chemistry on benthos.
Degree of Redox Potential Sulphides TOC Level1 Benthos Impact (mV) (µmol) Normal Normal >100 <300 Density and taxa richness [0 to 1.7%] ≥100% of reference Low impact or Slight increase 0 to 100 300 to 1300 Density ≥50% of reference, enrichment [1.7% to 5%] Richness >50% of reference Moderate to high Mod. increase -100 to 0 1300 to 6000 Density <50% of reference, impact [5% to 10%] Richness ≤10 Gross impact High TOC <-100 >6000 No macrofauna [>10%]
1 No quantitative information is given in the technical guidance as to what TOC values constitute "low", "moderate" or "gross" impact levels of TOC, except descriptive words. The values in brackets were selected by the authors for use in this report. The values given for redox, sulphides and benthos impact levels are taken from the PPTGD (Poole et al. 1978, Hargrave et al. 1995, cited in Environment Canada 2005).
Table 6.14 provides an estimated level of impact for each station in the Crofton EEM Cycle Five benthic invertebrate survey, based on guidance presented above. For comparison, Table 6.13 shows impact estimates at the same stations for Cycle Three. In Cycle Five, TOC at near-field stations and CRB1A and CRB2, the stations closest to the outfalls, exceeded normal concentrations and were classified as low impact/enrichment. Station CRB5C also was classified as low impact/enrichment, although this was likely due to its naturally highly muddy nature (90% silt/clay).
According to redox potential, station CRB1A (closest to the outfalls) in Cycle Five was classified as moderately impacted, while the other two near-field stations were characterized as having low impact. Farther along the gradient, stations to both the south and northwest ranged between normal to low impact conditions according to redox potential.
Sulphides in Cycle Five did not exceed 300 μMol at any station; sediments throughout the study area were classified as normal according to this variable.
When benthic invertebrate metrics were used to evaluate impact at each station in Cycle Five, conditions ranged between normal and low impact/enrichment. At the two stations closest to the outfalls (CRB1A and CRB2), both invertebrate density and richness were characterized as normal.
Relative to Cycle Three (2003), impact grades in the Crofton area have improved. In 2003, according to TOC and sulphides, conditions near the outfalls ranged between low to impact/enrichment, and conditions within stations used to calculate the benthic reference mean were overall characterized as normal to moderately impacted. According to benthic invertebrate densities and richness, communities closest to the outfalls were normal, and ranged between normal and low impact/enrichment closer to the reference stations. According to redox potential, all stations in the study area during 2003 were grossly impacted.
Crofton EEM Cycle Five 6-39 Hatfield Table 6.13 Assessment of sediment quality and benthic community health at each station sampled for Crofton EEM Cycle Three, February 2003, based on “impact grades” provided in EEM Technical Guidance.1
Sediment quality Benthic invertebrates2 Sampling Total Redox Total Adult Number of station organic potential sulphides density adult taxa carbon Near-field area CRB1A Moderate Gross Low Normal Normal CRB2 Moderate Moderate Low Normal Normal CRB2A Moderate Gross Low Normal Normal Northwest stations CRB4A Moderate Gross Moderate Low Low CRB5A Low Gross Low (Reference) (Reference) CRB5B Low Gross Low (Reference) (Reference) CRB5C Low Gross Low (Reference) (Reference) South stations CRB3 Moderate Gross Low Normal Normal CRB6 Moderate Gross Low Normal Normal CRB6A Low Gross Normal (Reference) (Reference) CRB6B Low Gross Normal (Reference) (Reference) CRB7 Low Gross Low (Reference) (Reference)
1 Environment Canada (1998); assessment criteria appear in Table 6.12. 2 Data from stations CRB5A, CRB5B, CRB5C, CRB6A, CRB6B, and CRB7 were used to determine mean and range of “normal” (reference) benthic invertebrate values.
Crofton EEM Cycle Five 6-40 Hatfield Table 6.14 Assessment of sediment quality and benthic community health at each station sampled for Crofton EEM Cycle Five, March 2009, based on “impact grades” provided in EEM Technical Guidance.1
Sediment quality Benthic invertebrates2 Sampling Total Redox Total Adult Number of station organic potential sulphides density adult taxa carbon Near-field area CRB1A Low Moderate Normal Normal Normal CRB2 Low Low Normal Normal Normal CRB2A Normal Low Normal Normal Low Northwest stations CRB4A Normal Low Normal Normal Low CRB5A Normal Low Normal (Reference) (Reference) CRB5B Normal Normal Normal (Reference) (Reference) CRB5C Low Low Normal (Reference) (Reference) South stations CRB3 Normal Normal Normal Normal Normal CRB7 Normal Normal Normal (Reference) (Reference) CRB6A Normal Low Normal (Reference) (Reference) CRB6B Normal Low Normal (Reference) (Reference) CRB6 Normal Low Normal (Reference) (Reference)
1 Environment Canada (2005); assessment criteria appear in Table 6.12. 2 Data from stations CRB5A, CRB5B, CRB5C, CRB6, CRB6A, CRB6B, and CRB7 were used to determine mean and range of “normal” (reference) benthic invertebrate values.
6.4.3 Characterizing Cycle Five Effects
Historical monitoring studies of the seabed in the immediate vicinity of the Crofton outfall diffusers in the 1970s and 1980s reported thick deposits of fibre in the immediate vicinity of the Crofton outfalls (see Hatfield 1994). Sediments were reported to be highly anoxic and to support depauperate benthic communities (Jones and Ellis 1975, Petrie and Holman 1983). Near present-day stations CRB1A, CRB2 and CRB2A, Jones and Ellis (1975) found benthic communities containing only 2 to 11 taxa, characterized by the pollution-tolerant polychaetes Capitella capitata and Polydora sp. and a leptostracan crustacean, Nebalia (Epinebalia) pugettensis. They estimated this depauperate area extended at least 900 m beyond the effluent outfalls. Areas beyond this for approximately 1.5 km supported somewhat higher numbers of taxa, although fewer than the “normal” community identified further afield, and low diversity, with communities dominated by very high densities of tube-dwelling spionid and paraonid polychaetes.
A sediment coring study in 1992 found fibre beds extending along a northwest-southeast axis, from approximately 1.85 km northwest of the outfalls to 0.6 km southeast, and approximately 450 m offshore (CBR 1992); fibre
Crofton EEM Cycle Five 6-41 Hatfield of maximum depth was located approximately 100 m northwest of the effluent outfalls, near current EEM monitoring station CRB1A. Volatile solids, which includes organic carbon and other volatile residues; constituted from 24.8 to 89.6% of sediment samples in the immediate vicinity of the outfalls. The area around the Crofton outfalls exhibited a classic toxic or inhibitory response to effluent constituents and excessive organic enrichment, namely low richness and low evenness (i.e., community composition was dominated by a few hardy species). At approximately this time (i.e., 1992), the Crofton pulpmill began secondary treatment of its effluent, which resulted in large reductions in suspended solids and BOD discharged from the pulpmill.
Observations over the duration of EEM benthic and sediment quality studies, from 1995 to 2009, indicate that environmental quality in the immediate vicinity of the Crofton outfalls has continually improved. Concentrations of organic carbon near the outfalls declined over the five cycles of EEM, oxidative state has improved, and C:N ratio has decreased, suggesting that the amount of terrestrial carbon in sediment, likely representative of residual fibre mat, is declining as this mat decomposes and/or is dispersed by near-bottom currents.
In Cycle Four, an investigation of cause into the historical fibre mat near Crofton indicated that the thickest, most exposed portion of the mat continues to persist close to the outfalls, but that conditions will continue to improve as impacted sediments slowly become further buried by natural marine sediments.
In Cycle Three, both stations nearest the outfalls – i.e., CRB1A and CRB2 – exhibited improved sediment quality relative to Cycle Two, higher invertebrate densities than other stations (exceeding the prescribed effect size), but similar taxa richness, evenness and community composition. Communities near the mill in Cycle Three were dominated by Capitellid worms, an opportunistic species known to thrive in organically impacted or enriched sediments (Pearson and Rosenberg 1978).
Between Cycle Three and Cycle Five, although sediment quality has demonstrated continued improvements, benthic communities continue to demonstrate effects along the effluent exposure gradient. Closer to the outfalls, where sediments continue to be characterized by elevated levels of terrestrial organic matter (i.e., higher C:N ratios), higher carbon content, and lower redox potential, benthic communities exhibit higher densities, are dominated by Capitellid worms, and are statistically less similar to reference communities farther away. These observations are consistent with conclusions in Cycle Three that, although the historical fibre mat in the vicinity of the Crofton outfalls has decomposed to a sufficient extent that it no longer exerts a toxic (inhibitory) effect on benthic communities, the remaining organic matter contributes to a mild enrichment effect at stations close to the outfalls (Hatfield 2003). Such conditions contribute to greater densities of organisms, but do not detrimentally affect species richness or community composition.
Consistent improvements in environmental quality observed from EEM Cycle One to Cycle Five, and the results of the investigation of cause in Cycle Four, indicate that current effluent discharges are not negatively affecting benthic habitat quality or the ability of the benthic environment to naturally recover from historical impacts.
Crofton EEM Cycle Five 6-42 Hatfield 7.0 CONCLUSIONS
Based on the results of the Crofton EEM Cycle Five program, the following conclusions can be made:
Sublethal Toxicity of Effluent:
o No effect of effluent on survival or growth of topsmelt (Atherinops affinis) larvae was observed; effects on echinoderm fertilization were observed at a mean effluent concentration of 23.0% (IC25); and algal reproduction was affected at a mean effluent concentration of 1.55% (IC25); and
o Maximum potential zones of sublethal effect from the effluent discharge point are <6 m for survival of fish, 26 m for invertebrate fertilization, and 388 m for algal reproduction.
Dioxins in Fish Tissue:
o In 2009, total TEQ concentrations in crabs at four stations were below the guideline (24.4 pg/g hepatopancreas), and levels at the remaining three stations had declined relative to 2006.
Fish Survey:
o Lower densities of older, larger, heavier mussels were present closer to the mill, indicating greater survival, energy use, and energy storage in these organisms relative to those at the reference station;
o Reproductive energy use did not show any significant differences between near-field and reference mussels;
o In contrast to mussels, oysters in the near-field demonstrated lower survival, energy use, and energy storage than reference oysters. Oysters at the near-field station were significantly smaller and lighter, both in general and for their age, and were present in higher densities relative to other stations;
o Ratios of male versus female mussels were similar between the near-field and reference areas, and GSI did not demonstrate any significant difference between the two areas, indicating that sex did not play a role in influencing the significantly greater survival, energy use, and energy storage of mussels in the near-field;
o Mussels were older in the near-field, indicating that recent spawning and/or spat settlement was not as successful at the near-field station;
Crofton EEM Cycle Five 7-1 Hatfield o In contrast to mussels, the youngest oysters in the study were only found at the near-field station, and in terms of their size-at-age the near-field oysters were substantially smaller; and
o Coexistence of mussels and oysters can be difficult, with the larger oysters often colonizing suitable habitat at the expense of mussels; given the more favourable growing conditions observed for oysters at the reference station (i.e., higher temperature and DO), and the presence of larger oysters and smaller mussels at this station, it is possible that oyster populations have settled at the reference station at the expense of mussels.
Benthic Invertebrate Community Survey:
o Although conditions have improved since Cycle Three, benthic communities in Cycle Five continued to demonstrate effects along the effluent exposure gradient. Closer to the outfalls, benthic communities exhibited higher densities, were dominated by Capitellid worms, and were statistically less similar to reference communities farther away;
o Despite improvements in sediment quality since Cycle Three, closer to the outfalls sediments continued to be characterized by elevated levels of terrestrial organic matter (i.e., higher C:N ratios), higher levels of organic carbon, and lower redox potential;
o Sulphides appeared to be relatively similar between near-field and far-field stations, indicating that oxygen is still available to microbes in sediments throughout the study area;
o According to the impact grading scheme, Cycle Five near-field sediment quality ranged between normal to moderately impacted, and benthic communities were representative of normal to low impact/enrichment conditions; and
o As in Cycle Three, the Cycle Five benthic study demonstrated that, although the historical fibre mat in the vicinity of the Crofton outfalls has decomposed to a sufficient extent that it no longer exerts a toxic (inhibitory) effect on benthic communities, the remaining organic matter contributes to a mild enrichment effect at stations close to the outfalls.
Crofton EEM Cycle Five 7-2 Hatfield
9.0 REFERENCES
Aldrich, J.C. and M. Crowley. 1986. Condition and variability in Mytilus edulis (L.) from different habitats in Ireland. Aquaculture, 52 (4): 273 – 286.
Anderson, E. 1977. Zinc content and condition of Pacific oysters (Crassostrea gigas) in the Crofton area, 1977. Dobrocky Seatech Ltd. Rept. to BC Forest Products Ltd. Crofton Pulp and Paper Division.
Anwar, N.A., C.A. Richardson, and R. Seed. 1990. Age determination, growth rate and population structure of the horse mussel Modiolus modiolus. Journal of Marine Biological Assessment U.K. 70: 441-457.
BCMOE (BC Ministry of Environment) (formerly WLAP). 2002. Protocols for Marine Environmental Monitoring. BC Ministry of Environment (formerly Water, Land and Air Protection), September 5, 2002, 29 pp.
Bray, J.R. and J.T. Curtis. 1957. Ordination of the upland forest communities of southern Wisconsin. Ecol. Monogr. 27:325-349.
Brown, J.R. and E.B. Hardwick (1988) Influences of temperature, salinity, and available food upon suspended culture of the Pacific oyster, Crassostrea gigas. II. Condition factor and survival. Aquaculture 70: 253-267.
Cardaso, J.F.M.F., D. Langlet, J.F. Loff, A.R. Martins, J.I.J.Witte, P.T. Santos, H.W. van der Veer. 2007. Spatial variability in growth and reproduction of the Pacific oyster Crassostrea gigas (Thunberg, 1793) along the west European coast. Journal of Sea Research, 57(4): 303-315.
Cohen, J. 1988. Statistical Power Analysis for the Behavioral Sciences. Second Edition. Lawrence Erlbaum Associates. Hillsdale, New Jersey.
Cowles, D. 2005. Mytilus trossulus Gould, 1850. Key to invertebrates found at or near the Rosario Beach Marine Laboratory (a campus of Walla Walla University), Fidalgo Island, Anacortes, WA. Accessed at: http://www.wallawalla.edu/academics/departments/biology/rosario/inv erts/Mollusca/Bivalvia/Mytiloida/Mytilidae/Mytilus_trossulus.html
Diederich, S. 2005. Differential recruitment of introduced Pacific oysters and native mussels at the North Sea coast: coexistence possible? Journal of Sea Research, 53(4): 269-281.
Ellis, D.V., P. Gee and S. Cross. 1981. Recovery from zinc contamination in a stock of Pacific oysters Crassostrea gigas (Thunberg). Water Poll. Res. J. of Canada 15(4): 303-310.
Crofton EEM Cycle Five 9-1 Hatfield Environment Canada. 1998. Biological Test Method: toxicity tests using early life stage of salmonid fish (rainbow trout), EPS 1/RM/28 2nd Ed. Method Development and Application Section, Environmental Technology Centre, Environment Canada, Ottawa, ON.
Environment Canada. 2005. Pulp and paper technical guidance for aquatic environmental effects monitoring. Environment Canada, January 2005.
Faul, F. and E. Erdfelder. 1992. GPOWER: A priori, post-hoc, and compromise power analyses for MS-DOS (computer program). Bonn University, Department of Psychology, Bonn, Germany.
Gibbons, W.N. and K.R. Munkittrick. 1994. A sentinel monitoring framework for identifying fish population responses to industrial discharges. J. Ecosys. Health 3:227-237.
Government of Canada. 2005. Gazette Part II, Vol. 138, No. 10. Regulations Amending the Pulp and Paper Effluent Regulations. 4 May, 2004. SOR/DORS/2004-109.
Government of Canada. 2008. Regulations Amending the Pulp and Paper Effluent Regulations. Registered July 28, 2008. Canada Gazette Part II, Vol. 142, No. 16 SOR/DORS/2008-239 published August 6, 2008.
Hatfield (Hatfield Consultants). 1994. Crofton environmental effects monitoring (EEM) pre-design reference document. Prepared for Fletcher Challenge Canada Ltd., Crofton Pulp and Paper, by Hatfield Consultants, West Vancouver, BC.
Hatfield. 1997. Crofton environmental effects monitoring (EEM) Cycle One Interpretive Report. 3 Volumes. Prepared for Fletcher Challenge Canada Ltd., Crofton Pulp and Paper, by Hatfield Consultants, West Vancouver, BC.
Hatfield. 1998. Wild oyster condition survey. Addendum to Fletcher Challenge Canada Ltd. Crofton Pulp and Paper 1997 Receiving Environment Report, B.C. Waste Management Permit PE-00114. Prepared for Fletcher Challenge Canada Ltd., Crofton Pulp and Paper, by Hatfield Consultants, West Vancouver, BC.
Hatfield. 2000. Crofton environmental effects monitoring (EEM) Cycle Two interpretive report, 1997 to 2000. 2 Volumes. Prepared for Fletcher Challenge Canada, Crofton Pulp and Paper, by Hatfield Consultants, West Vancouver, BC.
Hatfield. 2004. Crofton environmental effects monitoring (EEM) Cycle Three design document. Prepared for NorskeCanada Ltd., Crofton Division, by Hatfield Consultants, West Vancouver, BC.
Crofton EEM Cycle Five 9-2 Hatfield Hatfield. 2007. Crofton Environmental Effects Monitoring (EEM) Cycle Four Interpretive Report. Prepared for Catalyst Paper Corporation, Crofton Division, by Hatfield Consultants.
Hatfield. 2009a. Crofton environmental effects monitoring (EEM) Cycle Five design document. Prepared for Catalyst Paper, Crofton Division by Hatfield, March 2009.
Hatfield. 2009b. Crofton dioxin/furan trend monitoring program, 2009. Prepared for Catalyst Paper, Crofton Division by Hatfield, November 2009.
His, E. and R. Robert. 1987. Comparative effects of two antifouling paints on the oyster Crassostrea gigas. Marine Biology 95: 83-86.
Jones, A.A. and D.V. Ellis. 1975. Benthic Community Composition in the Crofton Mill Receiving Area, 1974; and Recommendations on a Routine Monitoring Program for Benthic Assessment. Prepared for BC Forest Products Ltd. by Dobrocky Seatech Ltd., Victoria, BC.
Paine, M. 1998. Volume II: CANMET/MMSL-INTEMIN Manual on Statistical Analysis of Environmental Data. Prepared by: Paine, Ledge and Associates (PLA), North Vancouver, BC. May 6, 1998.
Pearson, T.H. and R. Rosenberg. 1978. Macrobenhic succession in relation to organic enrichment and pollution of the marine environment. Oceanogr. Mar. Biol. Ann. Rev. 16:229-311.
Peterman, R.M. 1990. Statistical power analysis can improve fisheries research and management. Can. J. Fish. Aquat. Sci. 47: 2-15.
Petrie, L. and N. Holman. 1983. PISCES IV Sumbersible Dives 1973-1982. Department of Environment, Environmental Protection Service. Regional Programme Report 83-20.
Quayle, D.B. 1969. The effect of kraft mill effluent on the Pacific oyster (Crassostrea gigas) with particular reference to Crofton, B.C. Fish. Res. Bd. Can. Ms. Rep. 765.
Quayle, D.B. 1988. Pacific oyster culture in British Columbia. Canadian Bulletin of Fisheries and Aquatic Sciences 218.
Richardson, C.A., D.J. Crisp and N.W. Runham. 1970. Tidally deposited growth bands in the shell of the common cockle Cerastoderma edule (L.). Malacologia 18: 277-290.
Crofton EEM Cycle Five 9-3 Hatfield Salazar, M.H. 2000. Caged mussel pilot study, Port Alice mill, Vancouver Island, EEM Program. Regional manuscript report MS 00-01. Prepared for: Environment Canada, Pacific and Yukon Region, North Vancouver, BC. Prepared by: M. H. Salazar, Applied Biomonitoring, Kirkland, WA. November 2000.
Smith, B. and J. B. Wilson. 1996. A consumer’s guide to evenness indices. OIKOS 76:70-82.
SPSS Inc. 2000. Statistics I. SPSS Inc. United States of America.
St. Jean, S.D., P. van Poppelen, K. Kim, F. Bishay, and S.C. Courtenay. 2003. Gonadosomatic index for caged blue mussels: Pacific and Atlantic comparison. Poster presentation at Aquatic Toxicity Workshop, September 28, 2003, by Environment Canada and Greater Vancouver Regional District.
Crofton EEM Cycle Five 9-4 Hatfield 10.0 GLOSSARY
Acute With reference to toxicity tests with fish, usually means an effect that happens within four to seven days, or an exposure of that duration. An acute effect could be mild or sublethal, if it were rapid.
ANCOVA Analysis of covariance. ANCOVA compares regression lines, testing for differences in either slopes or intercepts (adjusted means).
ANOVA Analysis of variance. An ANOVA tests for differences among levels of one or more factors. For example, individual sites are levels of the factor site. Two or more factors can be included in an ANOVA (e.g., site and year).
BEAST Benthic assessment of sediment. BEAST is a tool for evaluating the health of benthic invertebrate communities by using predictive models that relate site habitat attributes to an expected community, commonly referred to as a reference condition (see CABIN and RCA, below).
Benthos Organisms that inhabit the bottom substrates (sediments, debris, logs, macrophytes) of aquatic habitats for at least part of their life cycle. The term benthic is used as an adjective, as in benthic invertebrates.
BOD Biochemical oxygen demand. The test measures the oxygen utilized during a specified incubation period for the biochemical degradation of organic material and the oxygen used to oxidize inorganic material such as sulfides and ferrous iron. Usually conducted as a 5-day test (i.e., BOD5).
13C (permil) Carbon isotope (13C) ratio.
CABIN Canadian aquatic biomonitoring network. CABIN is a collaborative programme developed and maintained by Environment Canada to establish a network of reference sites (see RCA, below) available to all users interested in assessing the biological health of fresh water in Canada.
CL Confidence limits. A set of possible values within which the true value will lie with a specified level of probability.
Crofton EEM Cycle Five 10-1 Hatfield Colour True colour of water is the colour of a filtered water sample (and thus with turbidity removed), and results from materials which are dissolved in the water. These materials include natural mineral components such as iron and calcium carbonate, as well as dissolved organic matter such as humic acids, tannin, and lignin. Organic and inorganic compounds from industrial or agricultural uses may also add colour to water. As with turbidity, colour hinders the transmission of light through water, and thus "regulates" biological processes within the body of water.
Community A set of taxa coexisting at a specified spatial or temporal scale.
Concentration Units Concentration Units Abbreviation Units Table Parts per million ppm mg/kg or μg/g or mg/L Parts per billion ppb μg/kg or ng/g or μg/L Parts per trillion ppt ng/kg or pg/g or ng/L Parts per quadrillion ppq pg/kg or fg/g or pg/L
Condition Factor A measure of the plumpness or fatness of aquatic organisms. For oysters and mussels, values are based on the ratio of the soft tissue dry weight to the volume of the shell cavity. For fish, the condition factor is based on length-weight relationships.
Conductivity A numerical expression of the ability of an aqueous solution to carry an electric current. This ability depends on the presence of ions, their total concentration, mobility, valence and relative concentrations, and on the temperature of measurement.
Covariate An independent variable; a measurement taken on each experimental unit that predicts to some degree the final response to the treatment, but which is unrelated to the treatment (e.g., body size [covariate] included in the analysis to compare gonad weights of fish collected from reference and exposed areas).
Dioxins/Furans Polychlorinated dibenzo-para-dioxins (PCDDs) and dibenzofurans (PCDFs) are often simply called dioxins, although they are two separate groups of substances with similar effects. There are 210 different compounds, of which 17 are the most toxic.
Crofton EEM Cycle Five 10-2 Hatfield DO Dissolved oxygen, the gaseous oxygen in solution with water. At low concentrations it may become a limiting factor for the maintenance of aquatic life. It is normally measured in milligrams/litre, and is widely used as a criterion of receiving water quality. The level of dissolved oxygen which can exist in water before the saturation point is reached is primarily controlled by temperature, with lower temperatures allowing for more oxygen to exist in solution. Photosynthetic activity may cause the dissolved oxygen to exist at a level which is higher than this saturation point, whereas respiration may cause it to exist at a level which is lower than this saturation point. At high saturation, fish may contract gas bubble disease, which produces lesions in blood vessels and other tissues and subsequent physiological dysfunctions.
ECp A point estimate of the concentration of test material that causes a specified percentage effective toxicity (sublethal or lethal). In most instances, the ECp is statistically derived by analysis of an observed biological response (e.g., incidence of nonviable embryos or reduced hatching success) for various test concentrations after a fixed period of exposure. EC25 is used for the rainbow trout sublethal toxicity test.
Fecundity The number of eggs or offspring produced by a female.
Gonad A male or female organ producing reproductive cells or gametes (i.e., female ovum, male sperm). The male gonad is the testis, the female gonad is the ovary.
GSI Gonadosomatic Index. Calculated by expressing gonad weight as a percentage of whole body weight.
Hardness Total hardness is defined as the sum of the calcium and magnesium concentrations, both expressed as calcium carbonate, in milligrams per litre.
ICp A point estimate of the concentration of test material that causes a specified percentage impairment in a quantitative biological test which measures a change in rate, such as reproduction, growth, or respiration.
LC50 Median lethal concentration. The concentration of a substance that is estimated to kill half of a group of organisms. The
duration of exposure must be specified (e.g., 96-hour LC50).
LSI Liver Somatic Index. Calculated by expressing liver weight as a percent of whole body weight.
Crofton EEM Cycle Five 10-3 Hatfield Macroinvertebrates Those invertebrate (without backbone) animals that are visible to the eye and retained by a sieve with 500 µm mesh openings for freshwater, or 1,000 µm mesh openings for marine surveys (EEM methods).
δ15N (permil) Nitrogen isotope (15N) ratio.
Negative Control Material (e.g., water) that is essentially free of contaminants and of any other characteristics that could adversely affect the test organism. It is used to assess the "background response" of the test organism to determine the acceptability of the test using predefined criteria.
Organochlorine Chlorine that is attached to an organic molecule. The amount present is expressed as the weight of the chlorine. There are thousands of such substances, including some that are manufactured specifically as pesticides because of their toxicity.
pH A measure of the acid or alkaline nature of water or some other medium. Specifically, pH is the negative logarithm of the + hydronium ion (H30 ) concentration (or more precisely, activity). Practically, pH 7 represents a neutral condition in which the acid hydrogen ions balance the alkaline hydroxide ions. The pH of the water can have an important influence on the toxicity and mobility of chemicals in pulpmill effluents.
Plume The main pathway for dispersal of effluent within the receiving waters, prior to its complete mixing.
Population A group of organisms belonging to a particular species or taxon, found within a particular region, territory or sampling unit. A collection of organisms that interbreed and share a bounded segment of space.
ppt Parts per thousand.
Quality Assurance (QA) Refers to the externally imposed technical and management practices which ensure the generation of quality and defensible data commensurate with the intended use of the data; a set of operating principles that, if strictly followed, will produce data of known defensible quality.
Quality Control (QC) Specific aspect of quality assurance which refers to the internal techniques used to measure and assess data quality and the remedial actions to be taken when data quality objectives are not realized.
Crofton EEM Cycle Five 10-4 Hatfield RCA Reference condition approach. The key to assessing the condition of our waterways through CABIN is the use of the Reference Condition Approach. Reference sites are established based on minimal impacts by human use, and present users with a baseline for assessing potentially impaired sites. The reference sites represent as many different geographic regions and stream sizes as possible and are used to establish the type of community of organisms expected to occur in the range of natural habitat types present in regions covered by the CABIN network. Once the reference condition has been established, sites suspected of being impaired are sampled. Differences between the organisms found at the reference sites and the test- site indicate the extent, if any, of impairment at the site.
Redox Potential (Eh) In marine sediments, the measurement of reduction and oxidation by testing electron movement and, consequently, available oxygen. Reference Toxicant A chemical of quantified toxicity to test organisms, used to gauge the fitness, health, and sensitivity of a batch of test organisms. Resin Acids Any of a class of vegetable substances, composed chiefly of esters and ethers of organic acids, that occur as a sticky yellow or brown substance exuded on the bark of various plants and trees, such as the pine and fir.
Salinity A measure of the quantity of dissolved salts in seawater - in parts per thousand (ppt) by weight.
SD Standard deviation.
SE Standard error.
Secondary Treatment A stage of purification of a liquid waste in which micro- organisms decompose organic substances in the waste. In the process, the micro-organisms use oxygen. Oxygen usually is supplied by mechanical aeration and/or large surface area of treatment ponds (lagoons). Most secondary treatment also reduces toxicity.
Sentinel Species A monitoring species selected to be representative of the local receiving environment.
Stressor An environmental factor or group of factors eliciting a response by a community.
Sublethal A concentration or level that would not cause death. An effect that is not directly lethal.
Crofton EEM Cycle Five 10-5 Hatfield T4CDD 2,3,7,8-tetrachlorodibenzo-para-dioxin, the most toxic dioxin.
TEQ Toxic Equivalents.
TN Total nitrogen.
TOC Total organic carbon (TOC).
Total-TEQs TEQs are calculated by multiplying the concentration of each congener with its respective International Toxicity Equivalency Factor (ITEF), to normalize concentrations to the level that would be produced by an equivalent amount of 2,3,7,8-T4CDD, then summing all the concentrations.
TS Total sulphides.
TSS Total suspended solids (TSS) is a measurement of the oven dry weight of particles of matter suspended in the water which can be filtered through a standard filter paper with pore size of 0.45 micrometres.
Turbidity Turbidity in water is caused by the presence of matter such as clay, silt, organic matter, plankton, and other microscopic organisms that are held in suspension.
v/v volume/volume - used to define dilution ratios for two liquids.
Crofton EEM Cycle Five 10-6 Hatfield
APPENDICES
Appendix A1
Sublethal Toxicity Data and Calculations
Figure A1.1 Mean (± SD) percent mortality and average dry weight of topsmelt (Atherinops affinis) exposed to final effluent and control water, Catalyst Paper - Crofton Division, EEM Cycle Five.
September 4, 2007 (Summer 2007) April 16, 2007 (Winter 2007) March 31, 2008 (Winter 2008)
% Mortality Dry Weight % Mortality Dry Weight % Mortality Dry Weight
50 2.0 50 2.0 50 2.0
40 40 40 1.5 1.5 1.5
30 30 30 1.0 1.0 1.0 20 20 20
0.5 Dry Weight (mg)
0.5 Dry Weight (mg)
0.5 Dry Weight (mg) Mean Mortality(%) Mean Mortality(%) Mean Mortality(%) 10 10 10
0 0.0 0 0.0 0 0.0 0.0 6.3 12.5 25 50 100 0.0 6.3 12.5 25 50 100 0.0 6.3 12.5 25 50 100 Effluent Concentration (% v/v) Effluent Concentration (% v/v) Effluent Concentration (% v/v) Figure A1.2 Mean (± SD) number of echinoderm eggs fertilized when exposed to final effluent and control water, Catalyst Paper - Crofton Division, EEM Cycle Five.
May 7, 2007 (Winter 2007) October 9, 2007 (Summer 2007) March 31, 2008 (Winter 2008)
110 110 110 100 100 100 90 90 90 80 80 80 70 70 70 60 60 60 50 50 50 40 40 40 No. of cystocarps No. of cystocarps No. of cystocarps 30 30 30 20 20 20 10 10 10 0 0 0 0.0 6.3 12.5 25.0 50.0 100.0 0.0 6.3 12.5 25.0 50.0 100.0 0.0 6.3 12.5 25.0 50.0 100.0 Effluent Concentration (% v/v) Effluent Concentration (% v/v) Effluent Concentration (% v/v)
Strongylocentrotus purpuratus Dendraster excentricus Strongylocentrotus purpuratus
January 12, 2009 (Summer 2008) April 7, 2009 (Winter 2009) November 24, 2009 (Summer 2009)
110 110 110 100 100 100 90 90 90 80 80 80 70 70 70 60 60 60 50 50 50 40 40 40 No. of cystocarps No. of cystocarps 30 No. of cystocarps 30 30 20 20 20 10 10 10 0 0 0 0.0 6.3 12.5 25.0 50.0 100.0 0.0 6.3 12.5 25.0 50.0 100.0 0.0 0.8 1.6 3.1 6.3 12.5 25.0 50.0 100.0 Effluent Concentration (% v/v) Effluent Concentration (% v/v) Effluent Concentration (% v/v)
Strongylocentrotus purpuratus Strongylocentrotus purpuratus Strongylocentrotus purpuratus Figure A1.3 Mean (± SD) number of cystocarps produced by an alga (Champia parvula ) exposed to final effluent and control water, Catalyst Paper - Crofton Division, EEM Cycle Five.
May 16, 2007 (Winter 2007) September 4, 2007 (Summer 2007) March 31, 2008 (Winter 2008)
40 50 30
40 30 20 30
20 20 10 No. of cystocarps No. of cystocarps No. of cystocarps 10 10
0 0 0 0 0.8 2.7 8.9 29.7 99 0.0 0.8 2.7 8.9 29.7 99.0 0 0.8 2.7 8.9 29.7 99 Effluent Concentration (% v/v) Effluent Concentration (% v/v) Effluent Concentration (% v/v)
January 12, 2009 (Summer 2008) May 25, 2009 (Winter 2009) November 24, 2009 (Summer 2009) 60 80 40
70 50 60 30 40 50
30 40 20
30 20 No. of cystocarps No. of cystocarps No. of cystocarps 20 10 10 10
0 0 0 0 0.2 0.8 2.7 8.9 29.7 99 0 0.2 0.8 2.7 8.9 29.7 99 0 0.16 0.8 4 19.8 99 Effluent Concentration (% v/v) Effluent Concentration (% v/v) Effluent Concentration (% v/v) Table A1.1 Catalyst Paper, Crofton Division, Sublethal Effluent Toxicity Test Results, Cycle Five.
Effluent Test type Collection Flag LC50% Flag IC25% > Project Description (S=Survival, LC50 Lower LC50 Upper IC25 Lower IC25 Upper Testing Period Date Consultant/Laboratory Species tested > for greater LC50 % for greater IC25 % Number (final, cooling, G=Growth, 95%cI 95%cI 95%cI 95%cI yyyymmdd than 100% than 100% etc.) R=Reproduction) Summer 2006 pp1119 final 20070307 Cantest Ltd. Strongylocentrotus purpuratus R 10.50 10.00 11.30 Winter 2007 pp1119 final 20070413 Cantest Ltd. Atherinops affinis S > 100 Winter 2007 pp1119 final 20070413 Cantest Ltd. Atherinops affinis G > 100 Winter 2007 pp1119 final 20070507 Cantest Ltd. Strongylocentrotus purpuratus R 21.01 18.72 25.66 Winter 2007 pp1119 final 20070416 Saskatchewan Research Council Champia parvula R 2.43 1.75 14.22 Summer 2007 pp1119 final 20070904 Cantest Ltd. Atherinops affinis S > 100 Summer 2007 pp1119 final 20070904 Cantest Ltd. Atherinops affinis G > 100 Summer 2007 pp1119 final 20071009 Cantest Ltd. Dendraster excentricus R 47.14 37.63 56.28 Summer 2007 pp1119 final 20070904 Saskatchewan Research Council Champia parvula R 1.44 0.81 1.57 Winter 2008 pp1119 final 20080331 Cantest Ltd. Atherinops affinis S > 100 Winter 2008 pp1119 final 20080331 Cantest Ltd. Atherinops affinis G > 100 Winter 2008 pp1119 final 20080331 Cantest Ltd. Strongylocentrotus purpuratus R 30.08 27.26 32.93 Winter 2008 pp1119 final 20080331 Saskatchewan Research Council Champia parvula R 1.6 0.00 3.79 Summer 2008 pp1119 final 20090112 Cantest Ltd. Strongylocentrotus purpuratus R 22.3 18.7 29 Summer 2008 pp1119 final 20090112 Aquatox Testing and Consulting Inc. Champia parvula R 1.2 0.5 1.2 Winter 2009 pp1119 final 20090407 Cantest Ltd. Strongylocentrotus purpuratus R 55.5 54.2 56.9 Winter 2009 pp1119 final 20090525 Cantest Ltd. Champia parvula R 0.8 0.63 1.28 Summer 2009 pp1119 final 20091124 Cantest Ltd. Strongylocentrotus purpuratus R 4.05 - 6.67 Summer 2009 pp1119 final 20091124 Saskatchewan Research Council Champia parvula R 2.56 0 3.23
Bold = Cycle 4 results (provided for comparision) Table A1.2 Catalyst Paper, Crofton Division Pulpmill - Calculation of geomeans and potential zones of sublethal effect.
Fish Invertebrate Algae Survival Growth Fertilization Reproduction Cycle 1 Cycle 2 Cycle 3 Cycle 4 Cycle 5 Cycle 1 Cycle 2 Cycle 3 Cycle 4 Cycle 5 Cycle 1 Cycle 2 Cycle 3 Cycle 4 Cycle 5 Cycle 1 Cycle 2 Cycle 3 Cycle 4 Cycle 5 LC50 LC50 LC50 LC50 LC50 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 IC25 67 67 67 100 100 67.9 67 67 100 100 6.5 18.47 1.85 19.7 47.14 23.91 22.29 3.22 2.43 65.5 67 67 100 100 65.5 67 67 100 100 35.8 38.29 29.46 28.54 21.01 54.68 4.95 2.31 1.44 66.7 68 67 100 100 66.7 68 67 97.88 100 4.2 70 19.67 12.72 30.08 4.4 67.3 5.88 3.22 1.6 68.2 67 100 68.2 67 100 66.7 11.97 67 22.3 4.5 16.97 2.27 1.2 67 100 67 100 20.85 31.25 55.5 7.81 4 0.8 100 100 100 100 38.79 10.5 4.05 2.93 1.95 2.56 100 100 17.55 19.02 100 100 53.93 3.09
Geomean 66.84 67.33 77.86 100.00 100.00 67.07 67.33 77.86 99.64 100.00 15.98 36.72 17.74 23.23 23.03 4.4 44.5 7.9 2.7 1.5 SE 0.6 0.3 6.0 0.0 0.0 0.6 0.3 6.0 0.4 0.0 14.7 11.6 5.7 8.4 7.7 0.0 10.0 2.8 0.3 0.3 1% effluent zone (m) 600 Zone of potential effect (m) 9.0 8.9 7.7 6.0 6.0 8.9 8.9 7.7 6.0 6.0 37.5 16.3 33.8 25.8 26.0 134.8 13.5 76.2 218.9 387.6
Appendix A2
Bivalve Survey: ALS Tissue Analytical Reports
Certificate of Analysis HATFIELD CONSULTANTS LTD. Report Date: 28-JAN-10 12:22 (MT) ATTN: NARA HENDERSON Version: FINAL REV. 3 201 - 1571 BELLEVUE AVE.
WEST VANCOUVER BC V7V 1A6
Lab Work Order #: L745473 Date Received: 24-MAR-09
Project P.O. #: NOT SUBMITTED Job Reference: CR1327 Legal Site Desc: CROFTON EEM CYCLE 5 CofC Numbers: C093349
Other Information:
Comments: Three sets of samples CR04, CR03, and CR05 (40 samples in each set) were submitted for weighing and then compositing and Lipid Content analysis of the composites. Each sample has been weighed and dry weight reported as Total Weight. Then, 40 samples of each set were composited and Lipid Content of the composites reported in the following data tables. 28-JAN-10: Please note that this report has been revised since its initial approval. This report replaces and supersedes all previous revision. The Total Weights for samples identified by ALS as L745473-86, L745473-88, L745473-97, L745473-98, and L745473-99 have now been included in the report. All other data remains unchanged. 28-JAN-10: Please see your final report attached
______Natasha Markovic-Mirovic Account Manager
THIS REPORT SHALL NOT BE REPRODUCED EXCEPT IN FULL WITHOUT THE WRITTEN AUTHORITY OF THE LABORATORY. ALL SAMPLES WILL BE DISPOSED OF AFTER 30 DAYS FOLLOWING ANALYSIS. PLEASE CONTACT THE LAB IF YOU REQUIRE ADDITIONAL SAMPLE STORAGE TIME.
8081 Lougheed Hwy, Suite 100, Burnaby, BC V5A 1W9 Phone: +1 604 253 4188 Fax: +1 604 253 6700 www.alsglobal.com A Campbell Brothers Limited Company L745473 CONTD.... PAGE 2 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-1 L745473-2 L745473-3 L745473-4 L745473-5 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-01 CR04-02 CR04-03 CR04-04 CR04-05
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.325 0.152 0.130 0.096 0.094 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 3 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-6 L745473-7 L745473-8 L745473-9 L745473-10 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-06 CR04-07 CR04-08 CR04-09 CR04-10
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.070 0.078 0.124 0.166 0.095 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 4 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-11 L745473-12 L745473-13 L745473-14 L745473-15 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-11 CR04-12 CR04-13 CR04-14 CR04-15
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.130 0.105 0.057 0.076 0.102 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 5 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-16 L745473-17 L745473-18 L745473-19 L745473-20 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-16 CR04-17 CR04-18 CR04-19 CR04-20
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.059 0.094 0.079 0.064 0.056 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 6 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-21 L745473-22 L745473-23 L745473-24 L745473-25 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-21 CR04-22 CR04-23 CR04-24 CR04-25
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.058 0.051 0.053 0.048 0.092 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 7 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-26 L745473-27 L745473-28 L745473-29 L745473-30 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-26 CR04-27 CR04-28 CR04-29 CR04-30
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.050 0.053 0.052 0.032 0.045 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 8 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-31 L745473-32 L745473-33 L745473-34 L745473-35 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-31 CR04-32 CR04-33 CR04-34 CR04-35
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.011 0.044 0.045 0.048 0.046 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 9 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-36 L745473-37 L745473-38 L745473-39 L745473-40 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 17:00 17:00 17:00 17:00 17:00 Client ID CR04-36 CR04-37 CR04-38 CR04-39 CR04-40
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.039 0.030 0.026 0.030 0.034 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 10 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-41 L745473-42 L745473-43 L745473-44 L745473-45 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-01 CR03-02 CR03-03 CR03-04 CR03-05
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.229 0.071 0.088 0.212 0.174 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 11 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-46 L745473-47 L745473-48 L745473-49 L745473-50 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-06 CR03-07 CR03-08 CR03-09 CR03-10
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.101 0.195 0.271 0.151 0.141 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 12 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-51 L745473-52 L745473-53 L745473-54 L745473-55 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-11 CR03-12 CR03-13 CR03-14 CR03-15
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.118 0.138 0.149 0.207 0.299 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 13 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-56 L745473-57 L745473-58 L745473-59 L745473-60 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-16 CR03-17 CR03-18 CR03-19 CR03-20
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.154 0.112 0.132 0.133 0.172 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 14 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-61 L745473-62 L745473-63 L745473-64 L745473-65 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-21 CR03-22 CR03-23 CR03-24 CR03-25
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.061 0.143 0.138 0.108 0.035 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 15 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-66 L745473-67 L745473-68 L745473-69 L745473-70 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-26 CR03-27 CR03-28 CR03-29 CR03-30
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.147 0.137 0.088 0.124 0.066 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 16 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-71 L745473-72 L745473-73 L745473-74 L745473-75 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-31 CR03-32 CR03-33 CR03-34 CR03-35
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.094 0.035 0.178 0.094 0.119 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 17 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-76 L745473-77 L745473-78 L745473-79 L745473-80 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:30 15:30 15:30 15:30 15:30 Client ID CR03-36 CR03-37 CR03-38 CR03-39 CR03-40
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.111 0.098 0.073 0.082 0.110 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 18 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-81 L745473-82 L745473-83 L745473-84 L745473-85 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-01 CR05-02 CR05-03 CR05-04 CR05-05
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.313 0.345 0.395 0.222 0.252 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 19 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-86 L745473-87 L745473-88 L745473-89 L745473-90 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-06 CR05-07 CR05-08 CR05-09 CR05-10
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.249 0.377 0.294 0.235 0.310 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 20 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-91 L745473-92 L745473-93 L745473-94 L745473-95 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-11 CR05-12 CR05-13 CR05-14 CR05-15
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.335 0.319 0.296 0.277 0.155 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 21 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-96 L745473-97 L745473-98 L745473-99 L745473-100 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-16 CR05-17 CR05-18 CR05-19 CR05-20
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.214 0.235 0.266 0.222 0.116 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 22 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-101 L745473-102 L745473-103 L745473-104 L745473-105 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-21 CR05-22 CR05-23 CR05-24 CR05-25
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.188 0.266 0.430 0.245 0.190 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 23 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-106 L745473-107 L745473-108 L745473-109 L745473-110 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-26 CR05-27 CR05-28 CR05-29 CR05-30
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.289 0.258 0.357 0.297 0.201 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 24 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-111 L745473-112 L745473-113 L745473-114 L745473-115 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-31 CR05-32 CR05-33 CR05-34 CR05-35
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.195 0.260 0.269 0.230 0.132 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 25 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-116 L745473-117 L745473-118 L745473-119 L745473-120 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 20:00 20:00 20:00 20:00 20:00 Client ID CR05-36 CR05-37 CR05-38 CR05-39 CR05-40
Grouping Analyte TISSUE Field Tests Total Weight (g) 0.172 0.172 0.232 0.228 0.275 Aggregate Lipid Content (%) Organics L745473 CONTD.... PAGE 26 of 27 ALS LABORATORY GROUP ANALYTICAL REPORT 28-JAN-10 12:24
Sample ID L745473-121 L745473-122 L745473-123 Description Sampled Date Sampled Time Client ID CR04 COMP (01 - CR03 COMP (01 - CR05 COMP (01 - 40) 40) 40)
Grouping Analyte TISSUE Field Tests Total Weight (g) Aggregate Lipid Content (%) 6.4 7.3 11.0 Organics L745473 CONTD.... PAGE 27 of 27 Reference Information 28-JAN-10 12:24
Additional Comments for Sample Listed: Samplenum Matrix Report Remarks Sample Comments
Methods Listed (if applicable): ALS Test Code Matrix Test Description Analytical Method Reference(Based On)
LIPIDS-GRAV-VA Tissue Lipids in Tissue by Gravimetric AOAC METHOD 983.23 This analysis is carried out using procedures adapted from the Official Methods of Analysis of AOAC International, Method 983.23, 16th Edition, 3rd Revision, 1997. The procedure involves a solvent extraction of a subsample of the tissue using a combination of chloroform and methanol in the presence of an enzyme. The extract is then evaporated to dryness and the residue weighed to determine Lipid Content.
** Laboratory Methods employed follow in-house procedures, which are generally based on nationally or internationally accepted methodologies. The last two letters of the above ALS Test Code column indicate the laboratory that performed analytical analysis for that test. Refer to the list below:
Laboratory Definition Code Laboratory Location Laboratory Definition Code Laboratory Location
VA ALS LABORATORY GROUP - VANCOUVER, BC, CANADA
GLOSSARY OF REPORT TERMS Surr - A surrogate is an organic compound that is similar to the target analyte(s) in chemical composition and behavior but not normally detected in enviromental samples. Prior to sample processing, samples are fortified with one or more surrogate compounds. The reported surrogate recovery value provides a measure of method efficiency. mg/kg (units) - unit of concentration based on mass, parts per million mg/L (units) - unit of concentration based on volume, parts per million N/A - Result not available. Refer to qualifier code and definition for explanation
Test results reported relate only to the samples as received by the laboratory. UNLESS OTHERWISE STATED, ALL SAMPLES WERE RECEIVED IN ACCEPTABLE CONDITION. Although test results are generated under strict QA/QC protocols, any unsigned test reports, faxes, or emails are considered preliminary.
ALS Laboratory Group has an extensive QA/QC program where all analytical data reported is analyzed using approved referenced procedures followed by checks and reviews by senior managers and quality assurance personnel. However, since the results are obtained from chemical measurements and thus cannot be guaranteed, ALS Laboratory Group assumes no liability for the use or interpretation of the results.
Certificate of Analysis HATFIELD CONSULTANTS LTD. Report Date: 04-SEP-09 16:54 (MT) ATTN: NOAH BAKER Version: FINAL 201 - 1571 BELLEVUE AVE.
WEST VANCOUVER BC V7V 1A6
Lab Work Order #: L800631 Date Received: 04-AUG-09
Project P.O. #: Job Reference: Legal Site Desc: CofC Numbers:
Other Information:
Comments: Three sets of samples, CR03, CR04, and CR05 (40 samples in each set) were submitted for weighing and then composited and Lipid Content analysis of the composites. Each sample has been dried and weighed and dry weight reported as Total Weight. 40 samples of each set were composited and Lipid Content of the composites reported in the following data tables.
______LINDSAY JONES Account Manager
THIS REPORT SHALL NOT BE REPRODUCED EXCEPT IN FULL WITHOUT THE WRITTEN AUTHORITY OF THE LABORATORY. ALL SAMPLES WILL BE DISPOSED OF AFTER 30 DAYS FOLLOWING ANALYSIS. PLEASE CONTACT THE LAB IF YOU REQUIRE ADDITIONAL SAMPLE STORAGE TIME.
1988 Triumph Street, Vancouver, BC V5L 1K5 Phone: +1 604 253 4188 Fax: +1 604 253 6700 www.alsglobal.com A Campbell Brothers Limited Company L800631 CONTD.... PAGE 2 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-1 L800631-2 L800631-3 L800631-4 L800631-5 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-1 CR03A-S-2 CR03A-S-3 CR03A-S-4 CR03A-S-5
Grouping Analyte TISSUE Field Tests Total Weight (g) 8.468 78.219 3.028 5.971 6.367 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 3 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-6 L800631-7 L800631-8 L800631-9 L800631-10 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-6 CR03A-S-7 CR03A-S-8 CR03A-S-9 CR03A-S-10
Grouping Analyte TISSUE Field Tests Total Weight (g) 5.59 6.026 3.267 4.251 4.41 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 4 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-11 L800631-12 L800631-13 L800631-14 L800631-15 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-11 CR03A-S-12 CR03A-S-13 CR03A-S-14 CR03A-S-15
Grouping Analyte TISSUE Field Tests Total Weight (g) 6.172 4.98 7.666 4.627 6.23 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 5 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-16 L800631-17 L800631-18 L800631-19 L800631-20 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-16 CR03A-S-17 CR03A-S-18 CR03A-S-19 CR03A-S-20
Grouping Analyte TISSUE Field Tests Total Weight (g) 1.86 5.432 5.43 4.581 8.181 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 6 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-21 L800631-22 L800631-23 L800631-24 L800631-25 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-21 CR03A-S-22 CR03A-S-23 CR03A-S-24 CR03A-S-25
Grouping Analyte TISSUE Field Tests Total Weight (g) 7.924 6.08 5.112 5.137 1.982 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 7 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-26 L800631-27 L800631-28 L800631-29 L800631-30 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-26 CR03A-S-27 CR03A-S-28 CR03A-S-29 CR03A-S-30
Grouping Analyte TISSUE Field Tests Total Weight (g) 2.985 4.251 1.963 1.35 3.968 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 8 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-31 L800631-32 L800631-33 L800631-34 L800631-35 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-31 CR03A-S-32 CR03A-S-33 CR03A-S-34 CR03A-S-35
Grouping Analyte TISSUE Field Tests Total Weight (g) 4.582 3.989 2.244 2.996 4.576 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 9 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-36 L800631-37 L800631-38 L800631-39 L800631-40 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-36 CR03A-S-37 CR03A-S-38 CR03A-S-39 CR03A-S-40
Grouping Analyte TISSUE Field Tests Total Weight (g) 3.679 2.631 6.155 4.218 8.171 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 10 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-41 L800631-42 L800631-43 L800631-44 L800631-45 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-41 CR03A-S-42 CR03A-S-43 CR03A-S-44 CR03A-S-45
Grouping Analyte TISSUE Field Tests Total Weight (g) 3.159 4.86 4.92 5.485 4.218 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 11 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-46 L800631-47 L800631-48 L800631-49 L800631-50 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-46 CR03A-S-47 CR03A-S-48 CR03A-S-49 CR03A-S-50
Grouping Analyte TISSUE Field Tests Total Weight (g) 3.934 4.655 5.026 2.872 1.058 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 12 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-51 L800631-52 L800631-53 L800631-54 L800631-55 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-51 CR03A-S-52 CR03A-S-53 CR03A-S-54 CR03A-S-55
Grouping Analyte TISSUE Field Tests Total Weight (g) 1.727 2.578 3.839 2.131 1.928 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 13 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-56 L800631-57 L800631-58 L800631-59 L800631-60 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-56 CR03A-S-57 CR03A-S-58 CR03A-S-59 CR03A-S-60
Grouping Analyte TISSUE Field Tests Total Weight (g) 7.728 5.196 3.195 3.767 4.996 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 14 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-61 L800631-62 L800631-63 L800631-64 L800631-65 Description Sampled Date 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 06-JUL-09 Sampled Time Client ID CR03A-S-61 CR03A-S-62 CR03A-S-63 CR03A-S-64 CR03A-S-65
Grouping Analyte TISSUE Field Tests Total Weight (g) 6.236 2.135 7.071 5.133 5.749 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 15 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-66 L800631-67 L800631-68 L800631-69 L800631-70 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-1 CR04A-S-2 CR04A-S-3 CR04A-S-4 CR04A-S-5
Grouping Analyte TISSUE Field Tests Total Weight (g) 8.948 18.838 16.431 10.874 12.707 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 16 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-71 L800631-72 L800631-73 L800631-74 L800631-75 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-6 CR04A-S-7 CR04A-S-8 CR04A-S-9 CR04A-S-10
Grouping Analyte TISSUE Field Tests Total Weight (g) 8.773 13.329 14.818 9.873 13.443 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 17 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-76 L800631-77 L800631-78 L800631-79 L800631-80 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-11 CR04A-S-12 CR04A-S-13 CR04A-S-14 CR04A-S-15
Grouping Analyte TISSUE Field Tests Total Weight (g) 7.505 10.972 9.823 4.531 15.695 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 18 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-81 L800631-82 L800631-83 L800631-84 L800631-85 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-16 CR04A-S-17 CR04A-S-18 CR04A-S-19 CR04A-S-20
Grouping Analyte TISSUE Field Tests Total Weight (g) 5.534 9.244 11.805 7.923 7.24 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 19 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-86 L800631-87 L800631-88 L800631-89 L800631-90 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-21 CR04A-S-22 CR04A-S-23 CR04A-S-24 CR04A-S-25
Grouping Analyte TISSUE Field Tests Total Weight (g) 2.787 11.415 5.061 10.209 4.953 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 20 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-91 L800631-92 L800631-93 L800631-94 L800631-95 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-26 CR04A-S-27 CR04A-S-28 CR04A-S-29 CR04A-S-30
Grouping Analyte TISSUE Field Tests Total Weight (g) 11.772 23.068 20.117 16.269 14.422 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 21 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-96 L800631-97 L800631-98 L800631-99 L800631-100 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-31 CR04A-S-32 CR04A-S-33 CR04A-S-34 CR04A-S-35
Grouping Analyte TISSUE Field Tests Total Weight (g) 9.94 12.412 14.847 16.868 12.768 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 22 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-101 L800631-102 L800631-103 L800631-104 L800631-105 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-36 CR04A-S-37 CR04A-S-38 CR04A-S-39 CR04A-S-40
Grouping Analyte TISSUE Field Tests Total Weight (g) 10.665 17.648 18.595 9.764 18.448 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 23 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-106 L800631-107 L800631-108 L800631-109 L800631-110 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-41 CR04A-S-42 CR04A-S-43 CR04A-S-44 CR04A-S-45
Grouping Analyte TISSUE Field Tests Total Weight (g) 19.065 16.485 13.808 15.471 15.401 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 24 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-111 L800631-112 L800631-113 L800631-114 L800631-115 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-46 CR04A-S-47 CR04A-S-48 CR04A-S-49 CR04A-S-50
Grouping Analyte TISSUE Field Tests Total Weight (g) 19.704 18.248 17.952 16.822 10.561 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 25 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-116 L800631-117 L800631-118 L800631-119 L800631-120 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-51 CR04A-S-52 CR04A-S-53 CR04A-S-54 CR04A-S-55
Grouping Analyte TISSUE Field Tests Total Weight (g) 7.992 14.898 3.554 5.649 6.825 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 26 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-121 L800631-122 L800631-123 L800631-124 L800631-125 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 Sampled Time Client ID CR04A-S-56 CR04A-S-57 CR04A-S-58 CR04A-S-59 CR04A-S-60
Grouping Analyte TISSUE Field Tests Total Weight (g) 9.909 13.14 7.161 8.469 8.343 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 27 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-126 L800631-127 L800631-128 L800631-129 L800631-130 Description Sampled Date 07-JUL-09 07-JUL-09 07-JUL-09 07-JUL-09 16-JUL-09 Sampled Time Client ID CR04A-S-61 CR04A-S-62 CR04A-S-63 CR04A-S-64 CR05A-S-1
Grouping Analyte TISSUE Field Tests Total Weight (g) 11.513 14.05 10.59 6.927 13.456 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 28 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-131 L800631-132 L800631-133 L800631-134 L800631-135 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-2 CR05A-S-3 CR05A-S-4 CR05A-S-5 CR05A-S-6
Grouping Analyte TISSUE Field Tests Total Weight (g) 16.477 12.829 11.686 14.674 11.025 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 29 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-136 L800631-137 L800631-138 L800631-139 L800631-140 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-7 CR05A-S-8 CR05A-S-9 CR05A-S-10 CR05A-S-11
Grouping Analyte TISSUE Field Tests Total Weight (g) 16.603 10.386 14.381 16.589 17.324 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 30 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-141 L800631-142 L800631-143 L800631-144 L800631-145 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-12 CR05A-S-13 CR05A-S-14 CR05A-S-15 CR05A-S-16
Grouping Analyte TISSUE Field Tests Total Weight (g) 19.473 8.108 16.275 16.777 11.194 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 31 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-146 L800631-147 L800631-148 L800631-149 L800631-150 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-17 CR05A-S-18 CR05A-S-19 CR05A-S-20 CR05A-S-21
Grouping Analyte TISSUE Field Tests Total Weight (g) 23.162 12.321 6.933 13.108 10.987 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 32 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-151 L800631-152 L800631-153 L800631-154 L800631-155 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-22 CR05A-S-23 CR05A-S-24 CR05A-S-25 CR05A-S-26
Grouping Analyte TISSUE Field Tests Total Weight (g) 15.966 15.878 12.147 15.887 2.897 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 33 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-156 L800631-157 L800631-158 L800631-159 L800631-160 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-27 CR05A-S-28 CR05A-S-29 CR05A-S-30 CR05A-S-31
Grouping Analyte TISSUE Field Tests Total Weight (g) 8.204 13.1 7.08 18.613 18.266 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 34 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-161 L800631-162 L800631-163 L800631-164 L800631-165 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-32 CR05A-S-33 CR05A-S-34 CR05A-S-35 CR05A-S-36
Grouping Analyte TISSUE Field Tests Total Weight (g) 18.46 16.056 17.341 10.977 5.667 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 35 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-166 L800631-167 L800631-168 L800631-169 L800631-170 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-37 CR05A-S-38 CR05A-S-39 CR05A-S-40 CR05A-S-41
Grouping Analyte TISSUE Field Tests Total Weight (g) 9.399 13.3 10.524 14.459 11.698 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 36 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-171 L800631-172 L800631-173 L800631-174 L800631-175 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-42 CR05A-S-43 CR05A-S-44 CR05A-S-45 CR05A-S-46
Grouping Analyte TISSUE Field Tests Total Weight (g) 20.956 14.36 15.721 17.719 16.951 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 37 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-176 L800631-177 L800631-178 L800631-179 L800631-180 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-47 CR05A-S-48 CR05A-S-49 CR05A-S-50 CR05A-S-51
Grouping Analyte TISSUE Field Tests Total Weight (g) 8.574 11.258 7.859 13.752 11.135 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 38 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-181 L800631-182 L800631-183 L800631-184 L800631-185 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-52 CR05A-S-53 CR05A-S-54 CR05A-S-55 CR05A-S-56
Grouping Analyte TISSUE Field Tests Total Weight (g) 4.538 10.179 14.703 8.004 6.368 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 39 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-186 L800631-187 L800631-188 L800631-189 L800631-190 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-57 CR05A-S-58 CR05A-S-59 CR05A-S-60 CR05A-S-61
Grouping Analyte TISSUE Field Tests Total Weight (g) 11.344 9.843 4.54 7.746 13.143 Aggregate Lipid Content (%) Organics L800631 CONTD.... PAGE 40 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-191 L800631-192 L800631-193 L800631-194 L800631-195 Description Sampled Date 16-JUL-09 16-JUL-09 16-JUL-09 Sampled Time Client ID CR05A-S-62 CR05A-S-63 CR05A-S-64 CR03A COMP (S1- CR04 COMP (S1- S65) S64)
Grouping Analyte TISSUE Field Tests Total Weight (g) 12.566 8.258 11.266 Aggregate Lipid Content (%) 12.6 14.4 Organics L800631 CONTD.... PAGE 41 of 42 ALS LABORATORY GROUP ANALYTICAL REPORT 04-SEP-09 16:56
Sample ID L800631-196 Description Sampled Date Sampled Time Client ID CR05A COMP (S1- S64)
Grouping Analyte TISSUE Field Tests Total Weight (g) Aggregate Lipid Content (%) 13.5 Organics L800631 CONTD.... PAGE 42 of 42 Reference Information 04-SEP-09 16:56
Additional Comments for Sample Listed: Samplenum Matrix Report Remarks Sample Comments
Methods Listed (if applicable): ALS Test Code Matrix Test Description Analytical Method Reference(Based On)
LIPIDS-GRAV-VA Tissue Lipids in Tissue by Gravimetric AOAC METHOD 983.23 This analysis is carried out using procedures adapted from the Official Methods of Analysis of AOAC International, Method 983.23, 16th Edition, 3rd Revision, 1997. The procedure involves a solvent extraction of a subsample of the tissue using a combination of chloroform and methanol in the presence of an enzyme. The extract is then evaporated to dryness and the residue weighed to determine Lipid Content.
** Laboratory Methods employed follow in-house procedures, which are generally based on nationally or internationally accepted methodologies. The last two letters of the above ALS Test Code column indicate the laboratory that performed analytical analysis for that test. Refer to the list below:
Laboratory Definition Code Laboratory Location Laboratory Definition Code Laboratory Location
VA ALS LABORATORY GROUP - VANCOUVER, BC, CANADA
GLOSSARY OF REPORT TERMS Surr - A surrogate is an organic compound that is similar to the target analyte(s) in chemical composition and behavior but not normally detected in enviromental samples. Prior to sample processing, samples are fortified with one or more surrogate compounds. The reported surrogate recovery value provides a measure of method efficiency. mg/kg (units) - unit of concentration based on mass, parts per million mg/L (units) - unit of concentration based on volume, parts per million N/A - Result not available. Refer to qualifier code and definition for explanation
Test results reported relate only to the samples as received by the laboratory. UNLESS OTHERWISE STATED, ALL SAMPLES WERE RECEIVED IN ACCEPTABLE CONDITION. Although test results are generated under strict QA/QC protocols, any unsigned test reports, faxes, or emails are considered preliminary.
ALS Laboratory Group has an extensive QA/QC program where all analytical data reported is analyzed using approved referenced procedures followed by checks and reviews by senior managers and quality assurance personnel. However, since the results are obtained from chemical measurements and thus cannot be guaranteed, ALS Laboratory Group assumes no liability for the use or interpretation of the results.
Appendix A3
Bivalve Survey: Metrics and Indices Data
Table A3.1 Whole-organism metrics and indices for Pacific blue mussels, Crofton EEM Cycle Five, March 2009.
Whole Wet tissue Dry Est. Cavity Condition Shell Size-at- Mussel Length Width Girth Mantle Shell wt. Age Station wet wt. wt. Sex tissue Volume Index Density GSI Age ID (mm) (mm) (mm) wt. (g) (g) (yr) (g) (g) wt. (g) (ml) (mg/cm3) (mg/cm3) (mg/yr) CRO3A 1 33 18 16 4.22 1.631 0.24 2.064 F 0.229 1.9008 120.5 1085.9 14.7 3.0 76.3 CRO3A 2 30 17 13 4.02 0.685 - 1.885 U 0.071 1.326 53.5 1421.6 3.0 23.7 CRO3A 3 25 14 12 2.48 0.57 0.045 1.141 M 0.088 0.84 104.8 1358.3 7.9 2.0 44.0 CRO3A 4 34 18 15 5.282 1.288 0.186 2.595 F 0.212 1.836 115.5 1413.4 14.4 3.0 70.7 CRO3A 5 33 19 14 3.671 1.11 0.103 1.911 M 0.174 1.7556 99.1 1088.5 9.3 3.0 58.0 CRO3A 6 32 17 15 4.41 0.825 - 1.993 U 0.101 1.632 61.9 1221.2 3.0 33.7 CRO3A 7 32 16 15 4.411 1.267 0.23 2.013 F 0.195 1.536 127.0 1310.5 18.2 3.0 65.0 CRO3A 8 39 20 17 5.635 1.771 0.295 3.057 F 0.271 2.652 102.2 1152.7 16.7 3.0 90.3 CRO3A 9 33 17 15 4.318 1.005 0.067 1.732 F 0.151 1.683 89.7 1029.1 6.7 3.0 50.3 CRO3A 10 28 15 13 2.821 0.853 0.23 1.288 M 0.141 1.092 129.1 1179.5 27.0 3.0 47.0 CRO3A 11 30 17 12 3.016 0.76 0.04 1.432 M 0.118 1.224 96.4 1169.9 5.3 3.0 39.3 CRO3A 12 30 16 8 3.586 0.93 0.158 1.5 F 0.138 0.768 179.7 1953.1 17.0 3.0 46.0 CRO3A 13 29 17 13 - 0.977 0.14 1.357 M 0.149 1.2818 116.2 1058.7 14.3 3.0 49.7 CRO3A 14 32 18 14 4.469 1.172 0.13 1.959 M 0.207 1.6128 128.3 1214.7 11.1 3.0 69.0 CRO3A 15 34 20 14 4.943 1.526 0.259 2.385 M 0.299 1.904 157.0 1252.6 17.0 3.0 99.7 CRO3A 16 29 16 13 2.981 0.979 0.153 1.298 F 0.154 1.2064 127.7 1075.9 15.6 3.0 51.3 CRO3A 17 29 16 11 2.253 0.78 0.09 1.1 M 0.112 1.0208 109.7 1077.6 11.5 3.0 37.3 CRO3A 18 30 17 14 4.248 0.975 0.085 1.709 F 0.132 1.428 92.4 1196.8 8.7 3.0 44.0 CRO3A 19 29 16 11 2.309 0.848 0.147 0.904 M 0.133 1.0208 130.3 885.6 17.3 3.0 44.3 CRO3A 20 31 16 13 2.485 1 0.23 1.305 F 0.172 1.2896 133.4 1011.9 23.0 3.0 57.3 CRO3A 21 31 17 14 3.788 0.499 - 1.618 U 0.061 1.4756 41.3 1096.5 3.0 20.3 CRO3A 22 27 17 12 2.78 0.838 0.102 1.145 M 0.143 1.1016 129.8 1039.4 12.2 3.0 47.7 CRO3A 23 27 14 13 2.747 0.887 0.165 1.128 M 0.138 0.9828 140.4 1147.7 18.6 3.0 46.0 CRO3A 24 28 15 12 3.066 0.677 0.064 1.489 F 0.108 1.008 107.1 1477.2 9.5 2.0 54.0 CRO3A 25 27 15 12 2.79 0.319 - 1.233 U 0.035 0.972 36.0 1268.5 3.0 11.7 CRO3A 26 27 17 13 2.629 0.958 0.099 1.131 M 0.147 1.1934 123.2 947.7 10.3 3.0 49.0 CRO3A 27 28 16 12 2.375 0.784 0.087 1.246 M 0.137 1.0752 127.4 1158.9 11.1 3.0 45.7 CRO3A 28 27 15 9 1.937 0.541 0.02 0.75 M 0.088 0.729 120.7 1028.8 3.7 3.0 29.3 CRO3A 29 26 16 11 2.345 0.737 0.112 0.925 F 0.124 0.9152 135.5 1010.7 15.2 3.0 41.3 CRO3A 30 26 14 11 1.571 0.448 - 0.895 U 0.066 0.8008 82.4 1117.6 3.0 22.0 CRO3A 31 25 15 11 2.195 0.666 0.06 0.978 M 0.094 0.825 113.9 1185.5 9.0 3.0 31.3 CRO3A 32 25 14 10 1.904 0.337 - 1.019 U 0.035 0.7 50.0 1455.7 3.0 11.7 CRO3A 33 29 16 12 2.113 0.987 0.211 0.991 M 0.178 1.1136 159.8 889.9 21.4 3.0 59.3 CRO3A 34 25 14 12 1.869 0.638 0.065 0.95 F 0.094 0.84 111.9 1131.0 10.2 3.0 31.3 CRO3A 35 29 16 11 2.46 0.692 0.063 1.008 F 0.119 1.0208 116.6 987.5 9.1 3.0 39.7 CRO3A 36 24 14 11 1.898 0.65 0.063 0.918 F 0.111 0.7392 150.2 1241.9 9.7 3.0 37.0 CRO3A 37 24 14 10 1.321 0.552 0.058 13671 F 0.098 0.672 145.8 998.5 10.5 3.0 32.7 CRO3A 38 24 13 11 1.625 0.496 0.038 0.876 F 0.073 0.6864 106.4 1276.2 7.7 3.0 24.3 CRO3A 39 24 13 10 1.592 0.49 0.045 0.706 M 0.082 0.624 131.4 1131.4 9.2 3.0 27.3 CRO3A 40 24 15 11 1.834 0.597 0.05 0.927 M 0.11 0.792 138.9 1170.5 8.4 3.0 36.7 CRO4 1 43 21 15 7.94 1.977 0.431 3.523 F 0.325 2.709 120.0 1300.5 21.8 4.0 81.3 CRO4 2 30 16 17 4.748 1.051 0.212 2.076 F 0.152 1.632 93.1 1272.1 20.2 3.0 50.7 CRO4 3 31 11 12 3.229 0.904 0.091 1.575 M 0.13 0.8184 158.8 1924.5 10.1 3.0 43.3 CRO4 4 31 15 13 3.333 0.626 0.076 1.564 F 0.096 1.209 79.4 1293.6 12.1 3.0 32.0 CRO4 5 29 16 11 2.216 0.582 0.047 1.291 F 0.094 1.0208 92.1 1264.7 8.1 3.0 31.3 CRO4 6 28 16 12 2.908 0.488 0.013 1.21 M 0.07 1.0752 65.1 1125.4 2.7 3.0 23.3 CRO4 7 25 14 10 1.416 0.484 0.044 0.674 M 0.078 0.7 111.4 962.9 9.1 2.0 39.0 CRO4 8 35 18 12 3.925 0.819 0.045 1.809 M 0.124 1.512 82.0 1196.4 5.5 3.0 41.3 CRO4 9 31 16 13 3.515 1.085 0.154 1.471 M 0.166 1.2896 128.7 1140.7 14.2 3.0 55.3 CRO4 10 29 14 11 2.124 0.61 0.066 0.725 F 0.095 0.8932 106.4 811.7 10.8 2.0 47.5 CRO4 11 33 16 12 3.006 0.814 0.07 1.285 M 0.13 1.2672 102.6 1014.0 8.6 2.0 65.0 CRO4 12 26 15 12 2.568 0.67 0.106 1.12 F 0.105 0.936 112.2 1196.6 15.8 2.0 52.5 CRO4 13 28 15 11 2.625 0.455 0.032 1.137 M 0.057 0.924 61.7 1230.5 7.0 3.0 19.0 CRO4 14 26 14 11 2.335 0.514 0.02 1.111 M 0.076 0.8008 94.9 1387.4 3.9 3.0 25.3 CRO4 15 28 14 11 2.68 0.609 0.058 0.912 F 0.102 0.8624 118.3 1057.5 9.5 2.0 51.0 CRO4 16 25 13 13 2.272 0.43 0.032 1.078 M 0.059 0.845 69.8 1275.7 7.4 3.0 19.7 CRO4 17 24 15 11 1.826 0.586 0.064 0.773 F 0.094 0.792 118.7 976.0 10.9 2.0 47.0 CRO4 18 27 14 10 1.986 0.535 0.06 0.821 M 0.079 0.756 104.5 1086.0 11.2 2.0 39.5 CRO4 19 26 13 10 1.65 0.475 - 0.405 U 0.064 0.676 94.7 599.1 0.0 2.0 32.0 CRO4 20 23 12 8 1.113 0.405 0.044 0.455 M 0.056 0.4416 126.8 1030.3 10.9 2.0 28.0 CRO4 21 24 13 10 1.806 0.399 0.032 0.797 F 0.058 0.624 92.9 1277.2 8.0 2.0 29.0 CRO4 22 27 13 11 1.947 0.389 0.022 0.847 M 0.051 0.7722 66.0 1096.9 5.7 3.0 17.0 CRO4 23 25 13 9 1.606 0.403 - 0.664 U 0.053 0.585 90.6 1135.0 0.0 2.0 26.5 CRO4 24 22 11 9 1.129 0.344 0.018 0.544 F 0.048 0.4356 110.2 1248.9 5.2 2.0 24.0 CRO4 25 24 12 11 1.527 0.504 0.06 0.792 F 0.092 0.6336 145.2 1250.0 11.9 3.0 30.7 CRO4 26 21 12 8 0.864 0.308 0.032 0.441 M 0.05 0.4032 124.0 1093.8 10.4 2.0 25.0 CRO4 27 22 12 9 1.087 0.333 0.022 0.525 M 0.053 0.4752 111.5 1104.8 6.6 2.0 26.5 CRO4 28 23 13 9 1.208 0.334 0.014 0.502 M 0.052 0.5382 96.6 932.7 4.2 2.0 26.0 CRO4 29 21 12 8 1.033 0.251 - 0.44 U 0.032 0.4032 79.4 1091.3 0.0 2.0 16.0
Page 1 of 2 Table A3.1 (Cont'd.)
Whole Wet tissue Dry Est. Cavity Condition Shell Size-at- Mussel Length Width Girth Mantle Shell wt. Age Station wet wt. wt. Sex tissue Volume Index Density GSI Age ID (mm) (mm) (mm) wt. (g) (g) (yr) (g) (g) wt. (g) (ml) (mg/cm3) (mg/cm3) (mg/yr) CRO4 30 21 12 9 1.121 0.321 0.031 0.496 M 0.045 0.4536 99.2 1093.5 9.7 2.0 22.5 CRO4 31 20 12 8 0.987 0.084 - 0.416 U 0.011 0.384 28.6 1083.3 0.0 1.0 11.0 CRO4 32 22 11 9 1.007 0.26 0.01 0.433 F 0.044 0.4356 101.0 994.0 3.8 2.0 22.0 CRO4 33 21 12 9 1.125 0.269 0.017 0.432 F 0.045 0.4536 99.2 952.4 6.3 2.0 22.5 CRO4 34 19 11 8 0.694 0.26 0.033 0.363 M 0.048 0.3344 143.5 1085.5 12.7 2.0 24.0 CRO4 35 20 12 8 0.988 0.291 0.014 0.465 F 0.046 0.384 119.8 1210.9 4.8 2.0 23.0 CRO4 36 18 10 8 0.926 0.236 0.021 0.485 F 0.039 0.288 135.4 1684.0 8.9 1.0 39.0 CRO4 37 18 10 8 0.775 0.228 0.019 0.363 M 0.03 0.288 104.2 1260.4 8.3 1.0 30.0 CRO4 38 18 10 7 0.64 0.186 0.006 0.276 F 0.026 0.252 103.2 1095.2 3.2 1.0 26.0 CRO4 39 17 10 7 0.636 0.185 0.011 - F 0.03 0.238 126.1 5.9 1.0 30.0 CRO4 40 18 10 8 0.655 0.191 0.013 - M 0.034 0.288 118.1 6.8 2.0 17.0 CRO5A 1 32 19 12 4.155 1.715 0.351 1.736 M 0.313 1.4592 214.5 1189.7 20.5 3.0 104.3 CRO5A 2 35 20 15 4.77 1.801 0.447 1.888 F 0.345 2.1 164.3 899.0 24.8 3.0 115.0 CRO5A 3 32 16 15 1.232 1.829 0.606 2.099 M 0.395 1.536 257.2 1366.5 33.1 3.0 131.7 CRO5A 4 32 17 13 3.83 1.143 0.159 1.779 F 0.222 1.4144 157.0 1257.8 13.9 3.0 74.0 CRO5A 5 31 17 12 3.317 1.2 0.208 1.358 M 0.252 1.2648 199.2 1073.7 17.3 3.0 84.0 CRO5A 6 31 16 14 3.44 1.134 0.226 1.72 F 0.249 1.3888 179.3 1238.5 19.9 3.0 83.0 CRO5A 7 35 19 14 4.57 1.868 0.435 1.914 M 0.377 1.862 202.5 1027.9 23.3 3.0 125.7 CRO5A 8 31 18 13 3.688 1.448 0.363 1.769 M 0.294 1.4508 202.6 1219.3 25.1 3.0 98.0 CRO5A 9 31 17 11 3.045 1.057 0.195 1.35 F 0.235 1.1594 202.7 1164.4 18.4 3.0 78.3 CRO5A 10 32 17 14 3.435 1.543 0.325 1.539 M 0.31 1.5232 203.5 1010.4 21.1 3.0 103.3 CRO5A 11 33 20 13 4.087 1.611 0.225 1.579 M 0.335 1.716 195.2 920.2 14.0 3.0 111.7 CRO5A 12 31 17 13 3.838 1.381 0.137 1.639 M 0.319 1.3702 232.8 1196.2 9.9 3.0 106.3 CRO5A 13 30 16 14 3.594 1.368 0.347 1.762 M 0.296 1.344 220.2 1311.0 25.4 3.0 98.7 CRO5A 14 28 15 14 3.075 1.268 0.311 1.376 M 0.277 1.176 235.5 1170.1 24.5 3.0 92.3 CRO5A 15 27 16 11 1.963 0.718 0.083 1.089 M 0.155 0.9504 163.1 1145.8 11.6 3.0 51.7 CRO5A 16 28 16 12 3.044 1.062 0.151 1.455 F 0.214 1.0752 199.0 1353.2 14.2 2.0 107.0 CRO5A 17 30 18 13 2.72 1.126 0.149 1.334 F 0.235 1.404 167.4 950.1 13.2 2.0 117.5 CRO5A 18 31 16 13 2.877 1.313 0.232 1.26 M 0.266 1.2896 206.3 977.0 17.7 3.0 88.7 CRO5A 19 31 17 12 3.08 1.197 0.225 1.332 M 0.222 1.2648 175.5 1053.1 18.8 3.0 74.0 CRO5A 20 24 14 10 1.52 0.518 0.079 1.55 M 0.116 0.672 172.6 2306.5 15.3 2.0 58.0 CRO5A 21 27 16 12 2.428 0.85 0.151 1.02 M 0.188 1.0368 181.3 983.8 17.8 3.0 62.7 CRO5A 22 29 14 12 3.355 1.14 0.142 1.736 M 0.266 0.9744 273.0 1781.6 12.5 3.0 88.7 CRO5A 23 38 20 14 5.412 2.165 0.434 2.245 F 0.43 2.128 202.1 1055.0 20.0 3.0 143.3 CRO5A 24 30 17 11 2.767 1.15 0.17 1.254 F 0.245 1.122 218.4 1117.6 14.8 3.0 81.7 CRO5A 25 29 14 11 2.283 1.059 0.234 1.12 F 0.19 0.8932 212.7 1253.9 22.1 3.0 63.3 CRO5A 26 29 16 13 3.313 1.206 0.166 1.572 M 0.289 1.2064 239.6 1303.1 13.8 CRO5A 27 29 16 13 2.16 1.318 0.207 1.285 M 0.258 1.2064 213.9 1065.2 15.7 3.0 86.0 CRO5A 28 33 17 13 4.042 1.741 0.457 1.88 M 0.357 1.4586 244.8 1288.9 26.2 3.0 119.0 CRO5A 29 31 17 14 3.553 1.44 0.264 1.61 F 0.297 1.4756 201.3 1091.1 18.3 3.0 99.0 CRO5A 30 30 16 11 2.776 0.971 0.158 1.227 F 0.201 1.056 190.3 1161.9 16.3 3.0 67.0 CRO5A 31 28 17 11 2.554 0.926 0.145 1.156 M 0.195 1.0472 186.2 1103.9 15.7 3.0 65.0 CRO5A 32 29 16 13 3.151 1.277 0.261 1.368 F 0.26 1.2064 215.5 1134.0 20.4 3.0 86.7 CRO5A 33 30 16 12 3.389 1.39 0.37 1.516 M 0.269 1.152 233.5 1316.0 26.6 3.0 89.7 CRO5A 34 28 16 12 2.904 1.197 0.244 1.345 F 0.23 1.0752 213.9 1250.9 20.4 3.0 76.7 CRO5A 35 24 13 9 1.655 0.62 0.074 0.821 F 0.132 0.5616 235.0 1461.9 11.9 3.0 44.0 CRO5A 36 25 14 11 2.153 0.831 0.124 0.996 F 0.172 0.77 223.4 1293.5 14.9 2.0 86.0 CRO5A 37 27 15 12 2.716 0.897 0.132 1.403 M 0.172 0.972 177.0 1443.4 14.7 3.0 57.3 CRO5A 38 28 14 12 2.949 1.171 0.352 1.375 M 0.232 0.9408 246.6 1461.5 30.1 3.0 77.3 CRO5A 39 28 16 11 2.44 1.115 0.191 1.122 F 0.228 0.9856 231.3 1138.4 17.1 3.0 76.0 CRO5A 40 28 16 13 3.047 1.275 0.371 1.546 F 0.275 1.1648 236.1 1327.3 29.1 3.0 91.7
Page 2 of 2 Table A3.2 Whole-organism metrics and indices for Pacific oysters, Crofton EEM Cycle Five, July 2009.
Whole Whole Wet tissue Shell Cavity Dry meat Oyster Length Width Girth Shell wt. Condition Shell Size-at- Station wet wt. volume wt. Volume Volume wt. Age ID (mm) (mm) (mm) (g) Index Density Age (g) (ml) (g) (ml) (ml) (g) CRO3A 1 114 75 47 300.9 190 41.2 227.4 110 80 8.5 105.9 2.1 7.0 1.2 CRO3A 2 97 60 47 259.2 150 30.9 199 80 70 er er 2.5 6.0 CRO3A 3 105 58.5 45 261.8 120 19.9 202.2 90 30 3.0 100.9 2.2 7.0 0.4 CRO3A 4 129 75 34 336.1 200 42.7 247.8 180 20 6.0 298.6 1.4 7.0 0.9 CRO3A 5 115 67 53 427.7 225 35.4 342.6 200 25 6.4 254.7 1.7 4.0 1.6 CRO3A 6 78 59 36 217.4 120 28.1 122.1 95 25 5.6 223.6 1.3 4.0 1.4 CRO3A 7 106 58 56 385.7 200 30.7 334.6 150 50 6.0 120.5 2.2 7.0 0.9 CRO3A 8 75 50 47 179 90 18.8 144.9 50 40 3.3 81.7 2.9 7.0 0.5 CRO3A 9 82 54 44 167.7 100 23.8 129.1 65 35 4.3 121.5 2.0 3.0 1.4 CRO3A 10 96 75 52 322.6 160 38.7 252.8 95 65 4.4 67.8 2.7 7.0 0.6 CRO3A 11 94 66 49 307.5 180 31.2 253 100 80 6.2 77.2 2.5 8.0 0.8 CRO3A 12 99 61 35 172.3 90 25 133.2 40 50 5.0 99.6 3.3 5.0 1.0 CRO3A 13 102 64 45 242.4 140 41.2 175.4 75 65 7.7 117.9 2.3 5.0 1.5 CRO3A 14 103 58 41 279.1 130 29.4 225.4 95 35 4.6 132.2 2.4 5.0 0.9 CRO3A 15 107 69 42 229.5 125 34.3 167.8 65 60 6.2 103.8 2.6 4.0 1.6 CRO3A 16 65 40 26 57.1 20 9.8 43.4 5 15 1.9 124.0 8.7 3.0 0.6 CRO3A 17 110 67 43 265.2 150 36.9 212.9 90 60 5.4 90.5 2.4 6.0 0.9 CRO3A 18 105 56 43 263.4 125 30.4 203.3 90 35 5.4 155.1 2.3 4.0 1.4 CRO3A 19 102 69 44 220.4 140 26.3 192.1 65 75 4.6 61.1 3.0 5.0 0.9 CRO3A 20 109 67 46 236.6 125 39.8 177.1 60 65 8.2 125.9 3.0 5.0 1.6 CRO3A 21 111 77 50 326.5 180 45.9 261.5 100 80 7.9 99.1 2.6 6.0 1.3 CRO3A 22 110 66 45 221.6 115 30.8 158.6 60 55 6.1 110.5 2.6 6.0 1.0 CRO3A 23 116 60 44 296.6 150 32.2 227.9 70 80 5.1 63.9 3.3 3.0 1.7 CRO3A 24 99 68 37 151 60 28.4 109 25 35 5.1 146.8 4.4 4.0 1.3 CRO3A 25 70 51 25 63.1 30 11.3 46.2 20 10 2.0 198.2 2.3 3.0 0.7 CRO3A 26 82.5 50 34 73.7 40 16.7 57.6 30 10 3.0 298.5 1.9 3.0 1.0 CRO3A 27 88 56.3 40.9 168.4 90 24.8 128.9 50 40 4.3 106.3 2.6 4.0 1.1 CRO3A 28 53 34.3 23.3 32.5 20 8.5 22.3 10 10 2.0 196.3 2.2 3.0 0.7 CRO3A 29 57.5 43.5 30.5 88.5 55 14.1 69 30 25 1.4 54.0 2.3 5.0 0.3 CRO3A 30 87 52.5 36 116.6 65 19.9 90.5 35 30 4.0 132.3 2.6 5.0 0.8 CRO3A 31 97 50.5 44 177 100 27.4 134.5 55 45 4.6 101.8 2.4 5.0 0.9 CRO3A 32 81 49 38.9 120.1 65 21.5 90.4 40 25 4.0 159.6 2.3 4.0 1.0 CRO3A 33 69.5 46 28 66.3 30 12.8 49.9 20 10 2.2 224.4 2.5 4.0 0.6 CRO3A 34 78 42.5 32 83.3 45 16.6 60 25 20 3.0 149.8 2.4 3.0 1.0 CRO3A 35 87.5 54 34 111.6 60 22.6 82.3 30 30 4.6 152.5 2.7 4.0 1.1 CRO3A 36 90 45 39 115.3 60 21 88 40 20 3.7 184.0 2.2 5.0 0.7 CRO3A 37 87 50 41 182.6 100 23.3 143.2 60 40 2.6 65.8 2.4 4.0 0.7 CRO3A 38 115 67 40 196.4 100 37.5 147.5 45 55 6.2 111.9 3.3 5.0 1.2 CRO3A 39 120 64 50 315.7 180 35.8 245 100 80 4.2 52.7 2.5 5.0 0.8 CRO3A 40 111 79 45 264 150 42.5 202.6 80 70 8.2 116.7 2.5 4.0 2.0 CRO3A 41 91 48 42 163.9 90 22.8 126.9 40 50 3.2 63.2 3.2 7.0 0.5 CRO3A 42 91 51 53 202.5 100 32.1 160.6 55 45 4.9 108.0 2.9 8.0 0.6 CRO3A 43 91 58 49 186.5 100 28.5 144.6 50 50 4.9 98.4 2.9 6.0 0.8 CRO3A 44 100 67 48 273.3 130 34 207.9 65 65 5.5 84.4 3.2 4.0 1.4 CRO3A 45 117 57 44 262.4 165 33.5 219.2 95 70 4.2 60.3 2.3 6.0 0.7 CRO3A 46 98 60 52 284.2 130 19.8 244.5 100 30 3.9 131.1 2.4 6.0 0.7 CRO3A 47 91 56 41 170.4 95 25.9 128.2 50 45 4.7 103.4 2.6 5.0 0.9 CRO3A 48 120 47 39 179.1 95 26.6 141.8 50 45 5.0 111.7 2.8 9.0 0.6 CRO3A 49 95 52.5 38 124 70 19.6 94.8 40 30 2.9 95.7 2.4 4.0 0.7 CRO3A 50 43 32 22 25.2 20 5.2 18.1 10 10 1.1 105.8 1.8 2.0 0.5 CRO3A 51 74 50 19 46.3 25 8.7 36.3 15 10 1.7 172.7 2.4 3.0 0.6 CRO3A 52 67 51 24 79.9 50 14.6 57.8 30 20 2.6 128.9 1.9 3.0 0.9 CRO3A 53 83.5 53 45 140.7 80 24.5 105.5 50 30 3.8 128.0 2.1 4.0 1.0 CRO3A 54 63 49 23 52 30 12.4 35.1 15 15 2.1 142.1 2.3 7.0 0.3 CRO3A 55 67 42 26 45.6 30 10.2 32.4 15 15 1.9 128.5 2.2 3.0 0.6 CRO3A 56 110 73 45 312.2 108 44.3 270.1 100 8 7.7 966.0 2.7 5.0 1.5 CRO3A 57 96 73 47 252.1 150 44.3 194.6 90 60 5.2 86.6 2.2 7.0 0.7 CRO3A 58 96 42 27 100.1 60 18.8 79 40 20 3.2 159.8 2.0 3.0 1.1 CRO3A 59 85 58 28 104.4 60 20.4 80.5 50 10 3.8 376.7 1.6 4.0 0.9 CRO3A 60 100 62 35 190.3 100 31.1 148.8 60 40 5.0 124.9 2.5 5.0 1.0 CRO3A 61 122 68 36 197.7 110 35.6 152.8 80 30 6.2 207.9 1.9 7.0 0.9 CRO3A 62 69 48 42 125.9 70 13.8 107.9 50 20 2.1 106.8 2.2 3.0 0.7 CRO3A 63 112 74 47 358.3 200 39.4 295.9 150 50 7.1 141.4 2.0 5.0 1.4 CRO3A 64 96 67 35 223.4 130 28.4 188.4 80 50 5.1 102.7 2.4 5.0 1.0 CRO3A 65 118 70 52 326.7 190 34.5 286.2 120 70 5.7 82.1 2.4 CRO4 1 96 77 44 197.5 125 45.7 130.3 55 70 8.9 127.8 2.4 4.0 2.2 CRO4 2 134 83 48 401.8 220 96.7 293.1 140 80 18.8 235.5 2.1 5.0 3.8 CRO4 3 117 85 45 268.3 150 69.5 182.5 65 85 16.4 193.3 2.8 6.0 2.7 er - lab error
Page 1 of 3 Table A3.2 (Cont'd.)
Whole Whole Wet tissue Shell Cavity Dry meat Oyster Length Width Girth Shell wt. Condition Shell Size-at- Station wet wt. volume wt. Volume Volume wt. Age ID (mm) (mm) (mm) (g) Index Density Age (g) (ml) (g) (ml) (ml) (g) CRO4 4 119 90 37 250.8 140 53.6 186.4 75 65 10.9 167.3 2.5 3.0 3.6 CRO4 5 123 88 35 260.5 145 53.9 186.4 75 70 12.7 181.5 2.5 5.0 2.5 CRO4 6 96 74 39 182.9 100 40.5 131.2 55 45 8.8 195.0 2.4 5.0 1.8 CRO4 7 103 78 35 293.6 150 62.5 199 65 85 13.3 156.8 3.1 4.0 3.3 CRO4 8 110 78 45 212 120 66.7 135.2 55 65 14.8 228.0 2.5 4.0 3.7 CRO4 9 133 60 39 201 110 49.7 149.2 60 50 9.9 197.5 2.5 3.0 3.3 CRO4 10 100 81 48 279.8 160 60.2 202.2 85 75 13.4 179.2 2.4 5.0 2.7 CRO4 11 87 78 30 164.9 105 35.4 127 60 45 7.5 166.8 2.1 3.0 2.5 CRO4 12 115 72 53 313.9 170 56.9 244.5 100 70 11.0 156.7 2.4 5.0 2.2 CRO4 13 120 75 42 298.8 160 52.7 221.5 90 70 9.8 140.3 2.5 7.0 1.4 CRO4 14 119 77 41 205.7 120 47.1 137.1 55 65 4.5 69.7 2.5 5.0 0.9 CRO4 15 138 102 44 349.4 200 74.4 247.1 100 100 15.7 157.0 2.5 4.0 3.9 CRO4 16 113 68 45 269.1 155 45.4 209.5 85 70 5.5 79.1 2.5 5.0 1.1 CRO4 17 105 68 37 201.1 100 40.3 153.7 50 50 9.2 184.9 3.1 3.0 3.1 CRO4 18 101 83 45 252.2 135 55.1 191.3 75 60 11.8 196.8 2.6 5.0 2.4 CRO4 19 100 76 39 203.8 110 42.1 149.8 60 50 7.9 158.5 2.5 4.0 2.0 CRO4 20 100 63 42 178.7 105 36.7 128.8 45 60 7.2 120.7 2.9 5.0 1.4 CRO4 21 135 59 39 186.9 105 46.4 116.3 40 65 2.8 42.9 2.9 4.0 0.7 CRO4 22 80 78 34 170.7 95 48.6 116 40 55 11.4 207.5 2.9 4.0 2.9 CRO4 23 89 58 33 110.2 55 25.8 77 30 25 5.1 202.4 2.6 4.0 1.3 CRO4 24 99 70 37 188.9 100 45.1 131.7 30 70 10.2 145.8 4.4 CRO4 25 107.5 70.5 35 152.1 100 35 103.9 50 50 5.0 99.1 2.1 5.0 1.0 CRO4 26 89 80 35 164.2 100 47.2 113.9 50 50 11.8 235.4 2.3 3.0 3.9 CRO4 27 128 88 37 337.7 200 90.3 242.7 100 100 23.1 230.7 2.4 5.0 4.6 CRO4 28 142.5 109 35.5 327.8 180 87.7 211.8 100 80 20.1 251.5 2.1 7.0 2.9 CRO4 29 163 95 35 435.1 210 77.4 343 120 90 16.3 180.8 2.9 7.0 2.3 CRO4 30 137 97 41 354.5 200 72.7 265.4 100 100 14.4 144.2 2.7 6.0 2.4 CRO4 31 151.5 59.5 45 331.3 190 67.5 24.1 100 90 9.9 110.4 0.2 6.0 1.7 CRO4 32 157 62.5 47 390.4 250 60.4 312.8 180 70 12.4 177.3 1.7 7.0 1.8 CRO4 33 200 75 35 323.7 180 66.9 240.5 100 80 14.8 185.6 2.4 4.0 3.7 CRO4 34 170 82 75 481.4 280 73.3 386.9 190 90 16.9 187.4 2.0 5.0 3.4 CRO4 35 180 70 61.5 560.2 300 85.2 440 200 100 12.8 127.7 2.2 6.0 2.1 CRO4 36 152 81 45 369 220 67.4 278 100 120 10.7 88.9 2.8 5.0 2.1 CRO4 37 124 90 38.5 260.2 150 68 185 100 50 17.6 353.0 1.9 7.0 2.5 CRO4 38 116 96 52 373.6 250 83.1 255.9 100 150 18.6 124.0 2.6 4.0 4.6 CRO4 39 120.5 79.5 41 310.8 200 52.7 233.5 100 100 9.8 97.6 2.3 4.0 2.4 CRO4 40 132 90 55 484.1 250 95.7 373.1 150 100 18.4 184.5 2.5 6.0 3.1 CRO4 41 124 89.5 37.5 358.7 200 77.3 252.3 100 100 19.1 190.7 2.5 4.0 4.8 CRO4 42 141 79 38 271.9 150 74.9 178.5 100 50 16.5 329.7 1.8 4.0 4.1 CRO4 43 124 71 34 238.7 110 59.6 159.4 70 40 13.8 345.2 2.3 6.0 2.3 CRO4 44 135 79 49 329.3 200 76.7 237.6 100 100 15.5 154.7 2.4 5.0 3.1 CRO4 45 134 92 50 345.7 200 63.4 267 100 100 15.4 154.0 2.7 8.0 1.9 CRO4 46 130 83 66 498.1 250 90 393.7 150 100 19.7 197.0 2.6 9.0 2.2 CRO4 47 135 93 43 516.2 300 90.1 394.2 180 120 18.2 152.1 2.2 5.0 3.7 CRO4 48 114 87 42 247 160 76.2 160.8 80 80 18.0 224.4 2.0 4.0 4.5 CRO4 49 135 86 36 282.7 165 71.7 200 85 80 16.8 210.3 2.4 5.0 3.4 CRO4 50 112 70 40 240.7 145 51.1 173.9 80 65 10.6 162.5 2.2 5.0 2.1 CRO4 51 91 70 30 155.2 80 36.7 110 35 45 8.0 177.6 3.1 4.0 2.0 CRO4 52 112 96 45 291.9 165 70.6 207.8 95 70 14.9 212.8 2.2 4.0 3.7 CRO4 53 80 63 32 103.6 60 23 74.9 30 30 3.6 118.5 2.5 4.0 0.9 CRO4 54 87 40 35 103.1 50 25.6 73.8 20 30 5.6 188.3 3.7 4.0 1.4 CRO4 55 85 70 33 105.8 60 29.3 67.8 20 40 6.8 170.6 3.4 4.0 1.7 CRO4 56 102 61 33 132.9 80 42.9 89 25 55 9.9 180.2 3.6 4.0 2.5 CRO4 57 125 77 46 293.7 170 70.4 210 95 75 13.1 175.2 2.2 5.0 2.6 CRO4 58 110 72 29 177.8 100 37.2 123 50 50 7.2 143.2 2.5 4.0 1.8 CRO4 59 85 80 40 138.1 80 39 97.3 45 35 8.5 242.0 2.2 4.0 2.1 CRO4 60 111 59 50 217.1 115 44.8 161.4 55 60 8.3 139.1 2.9 8.0 1.0 CRO4 61 115 72 29 248.5 280 63.9 170 90 190 11.5 60.6 1.9 4.0 2.9 CRO4 62 105 68 36 199 100 55 132.1 70 30 14.1 468.3 1.9 4.0 3.5 CRO4 63 105 73 44 268.7 180 49.8 206.5 90 90 10.6 117.7 2.3 5.0 2.1 CRO4 64 111 73 34 211.2 100 41.4 152.6 50 50 6.9 138.5 3.1 7.0 1.0 CRO5A 1 120 86 47 362.3 200 71.6 286.7 140 60 13.5 224.3 2.0 6.0 2.2 CRO5A 2 150 76 59 571.2 300 84 451.1 150 150 16.5 109.8 3.0 8.0 2.1 CRO5A 3 100 61 55 311.7 180 55.5 254.4 100 80 12.8 160.4 2.5 7.0 1.8 CRO5A 4 117 79 35 255.7 130 49 199.5 100 30 11.7 389.5 2.0 6.0 1.9 CRO5A 5 111 69 50 289.7 120 62.1 212.8 90 30 14.7 489.1 2.4 5.0 2.9 CRO5A 6 120 73 19 434.7 200 44.6 382.7 160 40 11.0 275.6 2.4 7.0 1.6 CRO5A 7 109 76 45 394.4 220 71.5 318.6 150 70 16.6 237.2 2.1 5.0 3.3 er - lab error
Page 2 of 3 Table A3.2 (Cont'd.)
Whole Whole Wet tissue Shell Cavity Dry meat Oyster Length Width Girth Shell wt. Condition Shell Size-at- Station wet wt. volume wt. Volume Volume wt. Age ID (mm) (mm) (mm) (g) Index Density Age (g) (ml) (g) (ml) (ml) (g) CRO5A 8 112 71 52 304.5 180 59.8 225.9 100 80 10.4 129.8 2.3 7.0 1.5 CRO5A 9 150 85 48 548.9 270 82.1 442.3 120 150 14.4 95.9 3.7 6.0 2.4 CRO5A 10 137 80 44 430.4 230 79.3 334.3 140 90 16.6 184.3 2.4 8.0 2.1 CRO5A 11 142 88 50 725.7 390 86.2 625.1 220 170 17.3 101.9 2.8 10.0 1.7 CRO5A 12 127 89 33 289.9 180 78.3 201.8 90 90 19.5 216.4 2.2 5.0 3.9 CRO5A 13 104 65 45 218.6 110 36.6 180.6 70 40 8.1 202.7 2.6 6.0 1.4 CRO5A 14 121 86 36 307 170 76.5 224.8 110 60 16.3 271.3 2.0 4.0 4.1 CRO5A 15 136 87 47 389.9 200 69.2 320.4 140 60 16.8 279.6 2.3 6.0 2.8 CRO5A 16 112 89 36 243.5 150 52.6 186.9 90 60 11.2 186.6 2.1 4.0 2.8 CRO5A 17 128 95 51 461.4 270 104.1 350.8 170 100 23.2 231.6 2.1 7.0 3.3 CRO5A 18 140 79 50 456.3 240 72 361.4 160 80 12.3 154.0 2.3 8.0 1.5 CRO5A 19 135 58 47 414.3 200 55.5 345.7 150 50 6.9 138.7 2.3 8.0 0.9 CRO5A 20 119 70 52 337.8 190 58.2 265.4 140 50 13.1 262.2 1.9 6.0 2.2 CRO5A 21 100 87 42 246.6 150 58.5 179.1 90 60 11.0 183.1 2.0 4.0 2.7 CRO5A 22 129 84 46 396.3 200 70.4 322.9 140 60 16.0 266.1 2.3 6.0 2.7 CRO5A 23 131 80 57 451.3 230 80.2 352.2 150 80 15.9 198.5 2.3 6.0 2.6 CRO5A 24 111 68 46 333.4 170 50.2 277.5 100 70 12.1 173.5 2.8 5.0 2.4 CRO5A 25 117 69 40 238.1 140 56.8 167.8 60 80 15.9 198.6 2.8 5.0 3.2 CRO5A 26 85 70 32 151.5 90 25.6 117.1 50 40 2.9 72.4 2.3 3.0 1.0 CRO5A 27 122 69 33 246 130 43.6 188.4 90 40 8.2 205.1 2.1 6.0 1.4 CRO5A 28 121 80 43 422.8 220 79.4 318.8 150 70 13.1 187.1 2.1 6.0 2.2 CRO5A 29 80 58 28 113.4 70 29.7 78.9 50 20 7.1 354.0 1.6 3.0 2.4 CRO5A 30 155 92 48 454.5 230 84.5 352.5 150 80 18.6 232.7 2.4 7.0 2.7 CRO5A 31 159 87 43 571.7 300 97.8 459.1 200 100 18.3 182.7 2.3 6.0 3.0 CRO5A 32 173 86 52 758.8 350 105.7 636.5 300 50 18.5 369.2 2.1 8.0 2.3 CRO5A 33 190 86 52 781.2 370 100 616.5 300 70 16.1 229.4 2.1 5.0 3.2 CRO5A 34 124 91 44 261.9 170 73.3 181 90 80 17.3 216.8 2.0 3.0 5.8 CRO5A 35 106 72 47 251.7 150 49.8 201.3 100 50 11.0 219.5 2.0 5.0 2.2 CRO5A 36 75 53 34 89 50 21.3 62.8 30 20 5.7 283.4 2.1 3.0 1.9 CRO5A 37 99 92 28 402.1 150 43.1 311.5 100 50 9.4 188.0 3.1 7.0 1.3 CRO5A 38 123 81 57 332.6 190 62.8 253.4 120 70 13.3 190.0 2.1 4.0 3.3 CRO5A 39 99 71 45 267.2 160 45.9 219 110 50 10.5 210.5 2.0 4.0 2.6 CRO5A 40 117 76 44 436.4 230 61.5 352.3 150 80 14.5 180.7 2.3 5.0 2.9 CRO5A 41 160 76 39 369.6 200 70.8 286.8 100 100 11.7 117.0 2.9 5.0 2.3 CRO5A 42 184 93 48 490.6 210 108.9 361.4 130 80 21.0 262.0 2.8 9.0 2.3 CRO5A 43 107 80 53 408.8 210 62.5 333.8 140 70 14.4 205.1 2.4 7.0 2.1 CRO5A 44 140 81 49 540.3 300 71.9 451.7 210 90 15.7 174.7 2.2 8.0 2.0 CRO5A 45 117 99 64 618.9 300 99.3 499.9 200 100 17.7 177.2 2.5 5.0 3.5 CRO5A 46 143 82 72 607.6 300 84.9 503 180 120 17.0 141.3 2.8 6.0 2.8 CRO5A 47 120 78 47 271.6 150 50 210.4 90 60 8.6 142.9 2.3 5.0 1.7 CRO5A 48 106 81 41 246 140 54.7 184 90 50 11.3 225.2 2.0 4.0 2.8 CRO5A 49 119 84 54 378.5 200 55.6 308 140 60 7.9 131.0 2.2 6.0 1.3 CRO5A 50 129 63 39 358.3 180 62.2 276.5 120 60 13.8 229.2 2.3 4.0 3.4 CRO5A 51 153 89 65 740.5 400 76 642.5 310 90 11.1 123.7 2.1 8.0 1.4 CRO5A 52 81 58 31 117.2 70 25.9 87.5 30 40 4.5 113.5 2.9 4.0 1.1 CRO5A 53 114 73 41 236.1 150 51 162.5 70 80 10.2 127.2 2.3 4.0 2.5 CRO5A 54 127 71 45 346.7 180 68.6 253.1 110 70 14.7 210.0 2.3 6.0 2.5 CRO5A 55 118 55 33 154.7 90 34.1 115.8 60 30 8.0 266.8 1.9 4.0 2.0 CRO5A 56 85 63 30 107.9 80 26.6 77.6 50 30 6.4 212.3 1.6 4.0 1.6 CRO5A 57 125 84 41 315.6 190 70.4 222.3 100 90 11.3 126.0 2.2 6.0 1.9 CRO5A 58 109 54 39 151.3 100 42.8 103.4 70 30 9.8 328.1 1.5 4.0 2.5 CRO5A 59 78 57 23 84.9 60 19.2 60.3 30 30 4.5 151.3 2.0 3.0 1.5 CRO5A 60 101 77 30 193.5 110 37.9 136.6 60 50 7.7 154.9 2.3 5.0 1.5 CRO5A 61 119 87 48 315.5 190 61.2 232.3 120 70 13.1 187.8 1.9 5.0 2.6 CRO5A 62 135 86 45 493 230 57.9 424.2 190 40 12.6 314.2 2.2 7.0 1.8 CRO5A 63 102 60 32 179.9 110 35.9 131.8 70 40 8.3 206.5 1.9 4.0 2.1 CRO5A 64 112 106 43 332.9 190 64.7 246.5 110 80 11.3 140.8 2.2 7.0 1.6 er - lab error
Page 3 of 3
Appendix A4
Tissue Survey: AXYS Sediment and Crab Analytical Reports
BATCH SUMMARY
Batch ID: WG28522 Date: 25-May-2009
Analysis Type: Dioxin/Furan Matrix Type: Tissue
BATCH MAKEUP
Contract: 2545 Blank: Samples: WG28522-101
L12548-1 CR1488-C1 L12548-2 CR1488-C2 L12548-3 CR1488-C3 L12548-4 CR1488-C4 L12548-5 CR1488-C7 L12548-6 CR1488-C8 Reference or Spike: L12548-7 CR1488-C17 WG28522-102
Duplicate:
Comments:
1. Data are not blank corrected.
Copyright AXYS Analytical Services Ltd February 1993
FQA-006 Rev. 2. 18-Jul-1994 BATCH SUMMARY
Batch ID: WG28385 Date: 08-May-2009
Analysis Type:Dioxin/Furan Matrix Type: Solid
BATCH MAKEUP
Contract: 2545 Blank: Samples: WG28385-101
L12478-1 CRDX09-S6
Reference or Spike: WG28385-102
Duplicate:
Comments:
1. Data are not blank corrected.
Copyright AXYS Analytical Services Ltd February 1993
FQA-006 Rev. 2. 18-Jul-1994 ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: CRDX09-S6 AXYS FILE: L12478-1 SAMPLE 10-Mar-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 08-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: SOLID EXTRACTION DATE: 03-Apr-2009 SAMPLE SIZE: 9.60 g (dry) INSTRUMENT: HR GC/MS ANALYSIS DATE: 29-Apr-2009 % Moisture: 52.2 CONCENTRATION IN: pg/g (dry weight basis)
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 2.29 0.10 1,2,3,7,8-PECDD 7.55 0.10 1,2,3,4,7,8-HXCDD 1.53 0.21 1,2,3,6,7,8-HXCDD 116 0.21 1,2,3,7,8,9-HXCDD 53.7 0.21 1,2,3,4,6,7,8-HPCDD 213 0.21 OCDD 1280 0.52 2,3,7,8-TCDF * 60.7 0.10 1,2,3,7,8-PECDF 1.31 0.10 2,3,4,7,8-PECDF 1.95 0.10 1,2,3,4,7,8-HXCDF 2.02 0.21 1,2,3,6,7,8-HXCDF 1.06 0.21 2,3,4,6,7,8-HXCDF 1.53 0.21 1,2,3,7,8,9-HXCDF NDR (0.21) 0.21 1,2,3,4,6,7,8-HPCDF 27.1 0.21 1,2,3,4,7,8,9-HPCDF 1.85 0.21 OCDF 73.1 0.52 TOTAL TETRA-DIOXINS 26.9 0.10 TOTAL PENTA-DIOXINS 63.3 0.10 TOTAL HEXA-DIOXINS 835 0.21 TOTAL HEPTA-DIOXINS 484 0.21 TOTAL TETRA-FURANS 123 0.10 TOTAL PENTA-FURANS 20.8 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 48.4 0.21 2,3,7,8-TCDD TEQs (ND=0) = 36.9 TOTAL HEPTA-FURANS 94.2 0.21 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 37.0
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 69.3 13C-1,2,3,7,8-PECDD 69.8 13C-1,2,3,6,7,8-HXCDD 69.7 13C-1,2,3,4,6,7,8-HPCDD 72.2 13C-OCDD 66.6 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 68.2 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 68.2 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 64.6 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 65.9 5. * = Concentration confirmed by analysis with DB-225 column
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Page 1 of 1 (WG28385 - AXYS_DIOXINS_AXYSDB5_L12478-1_SJ1005400.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: Lab Blank AXYS FILE: WG28385-101 SAMPLE N/A COLLECTION: CLIENT NO.: 2545 REPORT DATE: 08-May-2009 PROJECT NO.: N/A METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: SOLID EXTRACTION DATE: 03-Apr-2009 SAMPLE SIZE: 9.60 g INSTRUMENT: HR GC/MS ANALYSIS DATE: 29-Apr-2009 CONCENTRATION IN: pg/g
COMPOUND Concentration (SDL)
2,3,7,8-TCDD ND 0.10 1,2,3,7,8-PECDD ND 0.10 1,2,3,4,7,8-HXCDD ND 0.21 1,2,3,6,7,8-HXCDD ND 0.21 1,2,3,7,8,9-HXCDD ND 0.21 1,2,3,4,6,7,8-HPCDD ND 0.21 OCDD ND 0.52 2,3,7,8-TCDF ND 0.10 1,2,3,7,8-PECDF ND 0.10 2,3,4,7,8-PECDF ND 0.10 1,2,3,4,7,8-HXCDF ND 0.21 1,2,3,6,7,8-HXCDF ND 0.21 2,3,4,6,7,8-HXCDF ND 0.21 1,2,3,7,8,9-HXCDF ND 0.21 1,2,3,4,6,7,8-HPCDF ND 0.21 1,2,3,4,7,8,9-HPCDF ND 0.21 OCDF ND 0.52 TOTAL TETRA-DIOXINS ND 0.10 TOTAL PENTA-DIOXINS ND 0.10 TOTAL HEXA-DIOXINS ND 0.21 TOTAL HEPTA-DIOXINS ND 0.21 TOTAL TETRA-FURANS ND 0.10 TOTAL PENTA-FURANS ND 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS ND 0.21 2,3,7,8-TCDD TEQs (ND=0) = 0 TOTAL HEPTA-FURANS ND 0.21 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 0.20
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 67.2 13C-1,2,3,7,8-PECDD 68.3 13C-1,2,3,6,7,8-HXCDD 70.9 13C-1,2,3,4,6,7,8-HPCDD 73.1 13C-OCDD 63.6 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 66.1 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 70.2 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 70.8 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 70.8 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Jason MacKenzie______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 08-May-2009 10:32:18; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_WG28385-101_SJ1005406.html; Workgroup: WG28385; Design ID: 84 ]
Page 1 of 1 (WG28385 - AXYS_DIOXINS_AXYSDB5_WG28385-101_SJ1005406.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: Spiked Matrix AXYS FILE: WG28385-102
CLIENT NO.: 2545 REPORT DATE: 08-May-2009
SAMPLE TYPE: SOLID METHOD NO.: AXYS METHOD MLA-017 Rev 16
SAMPLE SIZE: 10.0 g INSTRUMENT: HR GC/MS
CONCENTRATION IN: pg/g
COMPOUND Determined Expected % Recovery
2,3,7,8-TCDD 10.4 10.6 97.9 1,2,3,7,8-PECDD 51.6 56.6 91.2 1,2,3,4,7,8-HXCDD 53.1 59.2 89.7 1,2,3,6,7,8-HXCDD 53.3 51.8 103 1,2,3,7,8,9-HXCDD 52.9 56.7 93.3 1,2,3,4,6,7,8-HPCDD 45.9 50.0 91.9 OCDD 95.8 108 88.8 2,3,7,8-TCDF 10.4 10.9 95.1 1,2,3,7,8-PECDF 43.2 50.0 86.4 2,3,4,7,8-PECDF 40.3 50.0 80.6 1,2,3,4,7,8-HXCDF 47.0 54.4 86.4 1,2,3,6,7,8-HXCDF 47.0 50.0 94.0 2,3,4,6,7,8-HXCDF 48.2 53.1 90.8 1,2,3,7,8,9-HXCDF 54.8 50.0 110 1,2,3,4,6,7,8-HPCDF 49.8 50.0 99.7 1,2,3,4,7,8,9-HPCDF 46.1 50.0 92.2 OCDF 98.8 109 91.0
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 65.0 13C-1,2,3,7,8-PECDD 62.0 1. Concentrations are recovery corrected 13C-1,2,3,6,7,8-HXCDD 61.9 13C-1,2,3,4,6,7,8-HPCDD 59.3 13C-OCDD 48.6 13C-2,3,7,8-TCDF 63.5 13C-1,2,3,7,8-PECDF 65.8 13C-1,2,3,4,7,8-HXCDF 63.4 13C-1,2,3,4,6,7,8-HPCDF 60.1
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Page 1 of 1 (WG28385 - AXYS_DIOXINS_AXYSDB5_WG28385-102_SJ1005402.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: CR1488-C1 AXYS FILE: L12548-1 SAMPLE 07-Apr-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 10.2 g (wet) INSTRUMENT: HR GC/MS ANALYSIS DATE: 01-May-2009 % Moisture: 78.6 CONCENTRATION IN: pg/g (wet weight basis) % Lipid: 15.1
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 1.65 0.10 1,2,3,7,8-PECDD 6.59 0.10 1,2,3,4,7,8-HXCDD 0.62 0.20 1,2,3,6,7,8-HXCDD 45.4 0.20 1,2,3,7,8,9-HXCDD 9.11 0.20 1,2,3,4,6,7,8-HPCDD 6.97 0.20 OCDD 4.18 0.49 2,3,7,8-TCDF * 37.7 0.10 1,2,3,7,8-PECDF 0.67 0.10 2,3,4,7,8-PECDF 1.82 0.10 1,2,3,4,7,8-HXCDF ND 0.20 1,2,3,6,7,8-HXCDF 0.33 0.20 2,3,4,6,7,8-HXCDF 0.44 0.20 1,2,3,7,8,9-HXCDF ND 0.20 1,2,3,4,6,7,8-HPCDF 2.12 0.20 1,2,3,4,7,8,9-HPCDF ND 0.20 OCDF ND 0.49 TOTAL TETRA-DIOXINS 4.77 0.10 TOTAL PENTA-DIOXINS 22.0 0.10 TOTAL HEXA-DIOXINS 171 0.20 TOTAL HEPTA-DIOXINS 12.0 0.20 TOTAL TETRA-FURANS 62.7 0.10 TOTAL PENTA-FURANS 12.2 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 9.23 0.20 2,3,7,8-TCDD TEQs (ND=0) = 18.3 TOTAL HEPTA-FURANS 3.16 0.20 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 18.3
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 73.8 13C-1,2,3,7,8-PECDD 74.9 13C-1,2,3,6,7,8-HXCDD 86.4 13C-1,2,3,4,6,7,8-HPCDD 79.7 13C-OCDD 66.9 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 73.5 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 69.8 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 85.4 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 82.0 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_L12548-1_SJ1007755.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_L12548-1_SJ1007755.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: CR1488-C2 AXYS FILE: L12548-2 SAMPLE 07-Apr-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 10.0 g (wet) INSTRUMENT: HR GC/MS ANALYSIS DATE: 01-May-2009 % Moisture: 77.0 CONCENTRATION IN: pg/g (wet weight basis) % Lipid: 13.6
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 1.64 0.10 1,2,3,7,8-PECDD 7.62 0.10 1,2,3,4,7,8-HXCDD 0.84 0.20 1,2,3,6,7,8-HXCDD 56.1 0.20 1,2,3,7,8,9-HXCDD 11.6 0.20 1,2,3,4,6,7,8-HPCDD 6.74 0.20 OCDD 3.94 0.50 2,3,7,8-TCDF * 40.1 0.10 1,2,3,7,8-PECDF 0.79 0.10 2,3,4,7,8-PECDF 1.99 0.10 1,2,3,4,7,8-HXCDF ND 0.20 1,2,3,6,7,8-HXCDF NDR (0.45) 0.20 2,3,4,6,7,8-HXCDF 0.46 0.20 1,2,3,7,8,9-HXCDF ND 0.20 1,2,3,4,6,7,8-HPCDF 2.62 0.20 1,2,3,4,7,8,9-HPCDF ND 0.20 OCDF 0.53 0.50 TOTAL TETRA-DIOXINS 7.74 0.10 TOTAL PENTA-DIOXINS 33.6 0.10 TOTAL HEXA-DIOXINS 213 0.20 TOTAL HEPTA-DIOXINS 11.8 0.20 TOTAL TETRA-FURANS 75.7 0.10 TOTAL PENTA-FURANS 18.4 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 13.1 0.20 2,3,7,8-TCDD TEQs (ND=0) = 20.9 TOTAL HEPTA-FURANS 4.08 0.20 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 20.9
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 78.6 13C-1,2,3,7,8-PECDD 75.3 13C-1,2,3,6,7,8-HXCDD 78.6 13C-1,2,3,4,6,7,8-HPCDD 80.4 13C-OCDD 70.5 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 73.5 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 73.1 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 77.6 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 73.9 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_L12548-2_SJ1007756.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_L12548-2_SJ1007756.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: CR1488-C3 AXYS FILE: L12548-3 SAMPLE 07-Apr-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 10.0 g (wet) INSTRUMENT: HR GC/MS ANALYSIS DATE: 01-May-2009 % Moisture: 77.7 CONCENTRATION IN: pg/g (wet weight basis) % Lipid: 10.7
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 2.08 0.10 1,2,3,7,8-PECDD 10.3 0.10 1,2,3,4,7,8-HXCDD 1.08 0.20 1,2,3,6,7,8-HXCDD 74.8 0.20 1,2,3,7,8,9-HXCDD 12.8 0.20 1,2,3,4,6,7,8-HPCDD 7.65 0.20 OCDD 3.22 0.50 2,3,7,8-TCDF * 55.9 0.10 1,2,3,7,8-PECDF 1.22 0.10 2,3,4,7,8-PECDF 3.43 0.10 1,2,3,4,7,8-HXCDF 1.03 0.20 1,2,3,6,7,8-HXCDF 0.54 0.20 2,3,4,6,7,8-HXCDF 0.72 0.20 1,2,3,7,8,9-HXCDF ND 0.20 1,2,3,4,6,7,8-HPCDF 3.29 0.20 1,2,3,4,7,8,9-HPCDF ND 0.20 OCDF ND 0.50 TOTAL TETRA-DIOXINS 9.12 0.10 TOTAL PENTA-DIOXINS 44.5 0.10 TOTAL HEXA-DIOXINS 259 0.20 TOTAL HEPTA-DIOXINS 14.0 0.20 TOTAL TETRA-FURANS 101 0.10 TOTAL PENTA-FURANS 25.4 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 19.5 0.20 2,3,7,8-TCDD TEQs (ND=0) = 28.2 TOTAL HEPTA-FURANS 4.20 0.20 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 28.3
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 77.9 13C-1,2,3,7,8-PECDD 78.4 13C-1,2,3,6,7,8-HXCDD 79.6 13C-1,2,3,4,6,7,8-HPCDD 72.7 13C-OCDD 73.4 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 74.4 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 74.9 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 75.8 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 77.5 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_L12548-3_SJ1007757.html; Workgroup: WG28522; Design ID: 84 ]
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CLIENT SAMPLE I.D.: CR1488-C4 AXYS FILE: L12548-4 SAMPLE 07-Apr-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 10.3 g (wet) INSTRUMENT: HR GC/MS ANALYSIS DATE: 01-May-2009 % Moisture: 80.6 CONCENTRATION IN: pg/g (wet weight basis) % Lipid: 9.19
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 2.59 0.10 1,2,3,7,8-PECDD 13.2 0.10 1,2,3,4,7,8-HXCDD 1.37 0.19 1,2,3,6,7,8-HXCDD 110 0.19 1,2,3,7,8,9-HXCDD 19.7 0.19 1,2,3,4,6,7,8-HPCDD 10.6 0.19 OCDD 3.02 0.48 2,3,7,8-TCDF * 47.7 0.10 1,2,3,7,8-PECDF 1.01 0.10 2,3,4,7,8-PECDF 2.35 0.10 1,2,3,4,7,8-HXCDF ND 0.19 1,2,3,6,7,8-HXCDF 0.60 0.19 2,3,4,6,7,8-HXCDF 0.64 0.19 1,2,3,7,8,9-HXCDF ND 0.19 1,2,3,4,6,7,8-HPCDF 2.97 0.19 1,2,3,4,7,8,9-HPCDF ND 0.19 OCDF ND 0.48 TOTAL TETRA-DIOXINS 9.96 0.10 TOTAL PENTA-DIOXINS 57.6 0.10 TOTAL HEXA-DIOXINS 397 0.19 TOTAL HEPTA-DIOXINS 17.6 0.19 TOTAL TETRA-FURANS 80.0 0.10 TOTAL PENTA-FURANS 22.0 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 16.2 0.19 2,3,7,8-TCDD TEQs (ND=0) = 34.7 TOTAL HEPTA-FURANS 3.39 0.19 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 34.7
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 76.8 13C-1,2,3,7,8-PECDD 77.9 13C-1,2,3,6,7,8-HXCDD 84.6 13C-1,2,3,4,6,7,8-HPCDD 93.1 13C-OCDD 71.3 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 74.1 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 74.8 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 80.6 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 80.1 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_L12548-4_SJ1007758.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_L12548-4_SJ1007758.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: CR1488-C7 AXYS FILE: L12548-5 SAMPLE 07-Apr-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 6.57 g (wet) INSTRUMENT: HR GC/MS ANALYSIS DATE: 01-May-2009 % Moisture: 83.5 CONCENTRATION IN: pg/g (wet weight basis) % Lipid: 6.41
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 1.44 0.15 1,2,3,7,8-PECDD 9.57 0.15 1,2,3,4,7,8-HXCDD 1.05 0.30 1,2,3,6,7,8-HXCDD 55.3 0.30 1,2,3,7,8,9-HXCDD 7.86 0.30 1,2,3,4,6,7,8-HPCDD 4.05 0.30 OCDD 1.33 0.76 2,3,7,8-TCDF * 37.3 0.15 1,2,3,7,8-PECDF 0.47 0.15 2,3,4,7,8-PECDF 2.43 0.15 1,2,3,4,7,8-HXCDF 0.68 0.30 1,2,3,6,7,8-HXCDF ND 0.30 2,3,4,6,7,8-HXCDF 0.49 0.30 1,2,3,7,8,9-HXCDF ND 0.30 1,2,3,4,6,7,8-HPCDF 1.25 0.30 1,2,3,4,7,8,9-HPCDF NDR (0.37) 0.30 OCDF ND 0.76 TOTAL TETRA-DIOXINS 7.20 0.15 TOTAL PENTA-DIOXINS 37.6 0.15 TOTAL HEXA-DIOXINS 179 0.30 TOTAL HEPTA-DIOXINS 6.93 0.30 TOTAL TETRA-FURANS 75.2 0.15 TOTAL PENTA-FURANS 22.1 0.15 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 10.8 0.30 2,3,7,8-TCDD TEQs (ND=0) = 22.1 TOTAL HEPTA-FURANS 1.25 0.30 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 22.1
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 79.1 13C-1,2,3,7,8-PECDD 83.0 13C-1,2,3,6,7,8-HXCDD 86.2 13C-1,2,3,4,6,7,8-HPCDD 84.7 13C-OCDD 80.9 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 77.2 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 78.1 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 82.8 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 85.1 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_L12548-5_SJ1007759.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_L12548-5_SJ1007759.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: CR1488-C8 AXYS FILE: L12548-6 SAMPLE 07-Apr-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 10.2 g (wet) INSTRUMENT: HR GC/MS ANALYSIS DATE: 01-May-2009 % Moisture: 72.6 CONCENTRATION IN: pg/g (wet weight basis) % Lipid: 17.6
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 2.54 0.10 1,2,3,7,8-PECDD 15.2 0.10 1,2,3,4,7,8-HXCDD 2.56 0.20 1,2,3,6,7,8-HXCDD 99.5 0.20 1,2,3,7,8,9-HXCDD 18.2 0.20 1,2,3,4,6,7,8-HPCDD 20.4 0.20 OCDD 9.10 0.49 2,3,7,8-TCDF * 52.6 0.10 1,2,3,7,8-PECDF 1.53 0.10 2,3,4,7,8-PECDF 4.08 0.10 1,2,3,4,7,8-HXCDF 2.19 0.20 1,2,3,6,7,8-HXCDF 1.00 0.20 2,3,4,6,7,8-HXCDF 1.15 0.20 1,2,3,7,8,9-HXCDF ND 0.20 1,2,3,4,6,7,8-HPCDF 5.65 0.20 1,2,3,4,7,8,9-HPCDF ND 0.20 OCDF 0.66 0.49 TOTAL TETRA-DIOXINS 11.2 0.10 TOTAL PENTA-DIOXINS 60.5 0.10 TOTAL HEXA-DIOXINS 354 0.20 TOTAL HEPTA-DIOXINS 32.2 0.20 TOTAL TETRA-FURANS 104 0.10 TOTAL PENTA-FURANS 41.0 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 36.2 0.20 2,3,7,8-TCDD TEQs (ND=0) = 37.0 TOTAL HEPTA-FURANS 6.95 0.20 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 37.0
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 66.6 13C-1,2,3,7,8-PECDD 72.7 13C-1,2,3,6,7,8-HXCDD 73.9 13C-1,2,3,4,6,7,8-HPCDD 70.4 13C-OCDD 61.0 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 63.9 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 67.2 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 65.9 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 69.1 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_L12548-6_SJ1007768.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_L12548-6_SJ1007768.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: CR1488-C17 AXYS FILE: L12548-7 SAMPLE 07-Apr-2009 COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: CR1488 METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 10.1 g (wet) INSTRUMENT: HR GC/MS ANALYSIS DATE: 01-May-2009 % Moisture: 78.3 CONCENTRATION IN: pg/g (wet weight basis) % Lipid: 10.1
COMPOUND Concentration (SDL)
2,3,7,8-TCDD 0.94 0.10 1,2,3,7,8-PECDD 4.04 0.10 1,2,3,4,7,8-HXCDD 0.56 0.20 1,2,3,6,7,8-HXCDD 24.1 0.20 1,2,3,7,8,9-HXCDD 5.38 0.20 1,2,3,4,6,7,8-HPCDD 3.71 0.20 OCDD 1.94 0.50 2,3,7,8-TCDF * 21.0 0.10 1,2,3,7,8-PECDF 0.46 0.10 2,3,4,7,8-PECDF 1.18 0.10 1,2,3,4,7,8-HXCDF 0.53 0.20 1,2,3,6,7,8-HXCDF 0.34 0.20 2,3,4,6,7,8-HXCDF 0.30 0.20 1,2,3,7,8,9-HXCDF ND 0.20 1,2,3,4,6,7,8-HPCDF 1.80 0.20 1,2,3,4,7,8,9-HPCDF ND 0.20 OCDF ND 0.50 TOTAL TETRA-DIOXINS 3.91 0.10 TOTAL PENTA-DIOXINS 13.9 0.10 TOTAL HEXA-DIOXINS 91.9 0.20 TOTAL HEPTA-DIOXINS 6.71 0.20 TOTAL TETRA-FURANS 38.6 0.10 TOTAL PENTA-FURANS 11.4 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS 9.36 0.20 2,3,7,8-TCDD TEQs (ND=0) = 10.6 TOTAL HEPTA-FURANS 1.80 0.20 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 10.6
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 67.0 13C-1,2,3,7,8-PECDD 69.6 13C-1,2,3,6,7,8-HXCDD 69.3 13C-1,2,3,4,6,7,8-HPCDD 66.1 13C-OCDD 62.9 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 66.0 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 63.2 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 69.9 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 70.4 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_L12548-7_SJ1007769.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_L12548-7_SJ1007769.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: Lab Blank AXYS FILE: WG28522-101 SAMPLE N/A COLLECTION: CLIENT NO.: 2545 REPORT DATE: 25-May-2009 PROJECT NO.: N/A METHOD NO.: AXYS METHOD MLA-017 Rev 16 SAMPLE TYPE: TISSUE EXTRACTION DATE: 17-Apr-2009 SAMPLE SIZE: 10.0 g INSTRUMENT: HR GC/MS ANALYSIS DATE: 30-Apr-2009 CONCENTRATION IN: pg/g
COMPOUND Concentration (SDL)
2,3,7,8-TCDD ND 0.10 1,2,3,7,8-PECDD ND 0.10 1,2,3,4,7,8-HXCDD ND 0.20 1,2,3,6,7,8-HXCDD ND 0.20 1,2,3,7,8,9-HXCDD ND 0.20 1,2,3,4,6,7,8-HPCDD ND 0.20 OCDD ND 0.50 2,3,7,8-TCDF ND 0.10 1,2,3,7,8-PECDF ND 0.10 2,3,4,7,8-PECDF ND 0.10 1,2,3,4,7,8-HXCDF ND 0.20 1,2,3,6,7,8-HXCDF ND 0.20 2,3,4,6,7,8-HXCDF ND 0.20 1,2,3,7,8,9-HXCDF ND 0.20 1,2,3,4,6,7,8-HPCDF ND 0.20 1,2,3,4,7,8,9-HPCDF ND 0.20 OCDF ND 0.50 TOTAL TETRA-DIOXINS ND 0.10 TOTAL PENTA-DIOXINS ND 0.10 TOTAL HEXA-DIOXINS ND 0.20 TOTAL HEPTA-DIOXINS ND 0.20 TOTAL TETRA-FURANS ND 0.10 TOTAL PENTA-FURANS ND 0.10 2,3,7,8-TCDD TEQs (Using WHO 2005 TEFs) TOTAL HEXA-FURANS ND 0.20 2,3,7,8-TCDD TEQs (ND=0) = 0 TOTAL HEPTA-FURANS ND 0.20 2,3,7,8-TCDD TEQs (ND=1/2 DL) = 0.19
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 78.9 13C-1,2,3,7,8-PECDD 84.8 13C-1,2,3,6,7,8-HXCDD 75.2 13C-1,2,3,4,6,7,8-HPCDD 76.3 13C-OCDD 67.2 1. SDL = Sample Detection Limit 13C-2,3,7,8-TCDF 76.9 2. ND = Not detected 3. NDR = peak detected but did not meet quantification criteria, result 13C-1,2,3,7,8-PECDF 76.1 reported represents the estimated maximum possible concentration 13C-1,2,3,4,7,8-HXCDF 79.9 4. Concentrations are recovery corrected 13C-1,2,3,4,6,7,8-HPCDF 80.2 5. * = Concentration confirmed by analysis with DB-225 column
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYS.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_WG28522-101_SJ1007762.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_WG28522-101_SJ1007762.html) ANALYSIS REPORT POLYCHLORINATED DIBENZODIOXINS AND DIBENZOFURANS AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811
CLIENT SAMPLE I.D.: Spiked Matrix AXYS FILE: WG28522-102
CLIENT NO.: 2545 REPORT DATE: 25-May-2009
SAMPLE TYPE: TISSUE METHOD NO.: AXYS METHOD MLA-017 Rev 16
SAMPLE SIZE: 10.0 g INSTRUMENT: HR GC/MS
CONCENTRATION IN: pg/g
COMPOUND Determined Expected % Recovery
2,3,7,8-TCDD 10.8 10.6 102 1,2,3,7,8-PECDD 53.8 56.6 95.1 1,2,3,4,7,8-HXCDD 56.3 59.2 95.1 1,2,3,6,7,8-HXCDD 55.9 51.8 108 1,2,3,7,8,9-HXCDD 56.0 56.7 98.7 1,2,3,4,6,7,8-HPCDD 48.2 50.0 96.5 OCDD 101 108 93.5 2,3,7,8-TCDF 10.7 10.9 98.2 1,2,3,7,8-PECDF 44.8 50.0 89.5 2,3,4,7,8-PECDF 45.7 50.0 91.4 1,2,3,4,7,8-HXCDF 48.3 54.4 88.9 1,2,3,6,7,8-HXCDF 48.5 50.0 97.0 2,3,4,6,7,8-HXCDF 54.7 53.1 103 1,2,3,7,8,9-HXCDF 58.9 50.0 118 1,2,3,4,6,7,8-HPCDF 51.2 50.0 102 1,2,3,4,7,8,9-HPCDF 49.8 50.0 99.6 OCDF 105 109 97.1
Surrogate Standards % Recovery
13C-2,3,7,8-TCDD 64.1 13C-1,2,3,7,8-PECDD 68.8 1. Concentrations are recovery corrected 13C-1,2,3,6,7,8-HXCDD 64.5 13C-1,2,3,4,6,7,8-HPCDD 63.9 13C-OCDD 64.4 13C-2,3,7,8-TCDF 61.5 13C-1,2,3,7,8-PECDF 59.9 13C-1,2,3,4,7,8-HXCDF 60.0 13C-1,2,3,4,6,7,8-HPCDF 63.9
Approved by: ______Shelley Facchin______QA/QC Chemist
For Axys Internal Use Only [ XSL Template: AXYSSpike.xsl; Created: 25-May-2009 11:28:14; Application: XMLTransformer-1.9.23; Report Filename: AXYS_DIOXINS_AXYSDB5_WG28522-102_SJ1007761.html; Workgroup: WG28522; Design ID: 84 ]
Page 1 of 1 (WG28522 - AXYS_DIOXINS_AXYSDB5_WG28522-102_SJ1007761.html)
Appendix A5
Benthic Survey: ALS Sediment Analytical Reports
Certificate of Analysis HATFIELD CONSULTANTS LTD. ATTN: NARA HENDERSON Reported On: 25-MAR-09 02:58 PM 201 - 1571 BELLEVUE AVE.
WEST VANCOUVER BC V7V 1A6
Lab Work Order #: L742011 Date Received: 13-MAR-09
Project P.O. #: Job Reference: CR1327-01 Legal Site Desc: CROFTON EEM SAMPLING CofC Numbers: 08-000536, 08-030490, 08-030494, 08-030495
Other Information:
Comments:
______NATASHA MARKOVIC-MIROVIC Account Manager
THIS REPORT SHALL NOT BE REPRODUCED EXCEPT IN FULL WITHOUT THE WRITTEN AUTHORITY OF THE LABORATORY. ALL SAMPLES WILL BE DISPOSED OF AFTER 30 DAYS FOLLOWING ANALYSIS. PLEASE CONTACT THE LAB IF YOU REQUIRE ADDITIONAL SAMPLE STORAGE TIME.
1988 Triumph Street, Vancouver, BC V5L 1K5 Phone: +1 604 253 4188 Fax: +1 604 253 6700 www.alsglobal.com A Campbell Brothers Limited Company L742011 CONTD.... PAGE 2 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-1 L742011-2 L742011-3 L742011-4 L742011-5 Description Sampled Date 10-MAR-09 10-MAR-09 10-MAR-09 10-MAR-09 10-MAR-09 Sampled Time 10:30 10:30 10:30 10:30 11:30 Client ID EEMCY5-CRB7-01 EEMCY5-CRB7-02 EEMCY5-CRB7-03 EEMCY5-CRB7-04 EEMCY5-CRB6-01
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) <1 % Sand (2.00mm - 1.00mm) (%) <1 % Sand (1.00mm - 0.50mm) (%) 4 % Sand (0.50mm - 0.25mm) (%) 22 % Sand (0.25mm - 0.125mm) (%) 43 % Sand (0.125mm - 0.063mm) (%) 21 % Silt (0.063mm - 0.0312mm) (%) 4 % Silt (0.0312mm - 0.004mm) (%) 3 % Clay (<4um) (%) 2 Leachable Anions Total Nitrogen by LECO (%) 0.04 0.04 0.04 0.07 & Nutrients Organic / Total Organic Carbon (%) 0.2 0.3 0.4 0.7 Inorganic Carbon L742011 CONTD.... PAGE 3 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-6 L742011-7 L742011-8 L742011-9 L742011-10 Description Sampled Date 10-MAR-09 10-MAR-09 10-MAR-09 10-MAR-09 10-MAR-09 Sampled Time 11:30 11:30 11:30 12:15 12:15 Client ID EEMCY5-CRB6-02 EEMCY5-CRB6-03 EEMCY5-CRB6-04 EEMCY5-CRB5C- EEMCY5-CRB5C- 01 02
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) 14 % Sand (2.00mm - 1.00mm) (%) 23 % Sand (1.00mm - 0.50mm) (%) 13 % Sand (0.50mm - 0.25mm) (%) 22 % Sand (0.25mm - 0.125mm) (%) 16 % Sand (0.125mm - 0.063mm) (%) 3 % Silt (0.063mm - 0.0312mm) (%) 5 % Silt (0.0312mm - 0.004mm) (%) 2 % Clay (<4um) (%) 3 Leachable Anions Total Nitrogen by LECO (%) 0.05 0.08 0.40 0.41 & Nutrients Organic / Total Organic Carbon (%) 0.4 0.9 3.5 3.6 Inorganic Carbon L742011 CONTD.... PAGE 4 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-11 L742011-12 L742011-13 L742011-14 L742011-15 Description Sampled Date 10-MAR-09 10-MAR-09 10-MAR-09 10-MAR-09 10-MAR-09 Sampled Time 12:15 12:15 14:00 14:00 14:00 Client ID EEMCY5-CRB5C- EEMCY5-CRB5C- EEMCY5-CRB5A- EEMCY5-CRB5A- EEMCY5-CRB5A- 03 04 01 02 03
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) <1 % Sand (2.00mm - 1.00mm) (%) 3 % Sand (1.00mm - 0.50mm) (%) 1 % Sand (0.50mm - 0.25mm) (%) <1 % Sand (0.25mm - 0.125mm) (%) 2 % Sand (0.125mm - 0.063mm) (%) 4 % Silt (0.063mm - 0.0312mm) (%) 35 % Silt (0.0312mm - 0.004mm) (%) 39 % Clay (<4um) (%) 16 Leachable Anions Total Nitrogen by LECO (%) 0.43 0.08 0.06 0.07 & Nutrients Organic / Total Organic Carbon (%) 3.6 0.5 0.4 0.5 Inorganic Carbon L742011 CONTD.... PAGE 5 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-16 L742011-17 L742011-18 L742011-19 L742011-20 Description Sampled Date 10-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 14:00 09:00 09:00 09:00 09:00 Client ID EEMCY5-CRB5A- EEMCY5-CRB2-01 EEMCY5-CRB2-02 EEMCY5-CRB2-03 EEMCY5-CRB2-04 04
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) 5 <1 % Sand (2.00mm - 1.00mm) (%) 5 2 % Sand (1.00mm - 0.50mm) (%) 9 3 % Sand (0.50mm - 0.25mm) (%) 31 16 % Sand (0.25mm - 0.125mm) (%) 31 36 % Sand (0.125mm - 0.063mm) (%) 7 19 % Silt (0.063mm - 0.0312mm) (%) 4 10 % Silt (0.0312mm - 0.004mm) (%) 5 9 % Clay (<4um) (%) 4 6 Leachable Anions Total Nitrogen by LECO (%) 0.12 0.11 0.12 & Nutrients Organic / Total Organic Carbon (%) 2.2 1.9 2.1 Inorganic Carbon L742011 CONTD.... PAGE 6 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-21 L742011-22 L742011-23 L742011-24 L742011-25 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 10:30 10:30 10:30 10:30 11:30 Client ID EEMCY5-CR5B-01 EEMCY5-CR5B-02 EEMCY5-CR5B-03 EEMCY5-CR5B-04 EEMCY5-CRB4A- 01
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) <1 % Sand (2.00mm - 1.00mm) (%) <1 % Sand (1.00mm - 0.50mm) (%) 4 % Sand (0.50mm - 0.25mm) (%) 30 % Sand (0.25mm - 0.125mm) (%) 50 % Sand (0.125mm - 0.063mm) (%) 7 % Silt (0.063mm - 0.0312mm) (%) 1 % Silt (0.0312mm - 0.004mm) (%) 3 % Clay (<4um) (%) 4 Leachable Anions Total Nitrogen by LECO (%) 0.04 0.06 0.06 0.10 & Nutrients Organic / Total Organic Carbon (%) 0.2 0.4 0.4 1.7 Inorganic Carbon L742011 CONTD.... PAGE 7 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-26 L742011-27 L742011-28 L742011-29 L742011-30 Description Sampled Date 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 11-MAR-09 Sampled Time 11:30 11:30 11:30 15:00 15:00 Client ID EEMCY5-CRB4A- EEMCY5-CRB4A- EEMCY5-CRB4A- EEMCY5-CRB3-01 EEMCY5-CRB3-02 02 03 04
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) 1 % Sand (2.00mm - 1.00mm) (%) <1 % Sand (1.00mm - 0.50mm) (%) 3 % Sand (0.50mm - 0.25mm) (%) 15 % Sand (0.25mm - 0.125mm) (%) 36 % Sand (0.125mm - 0.063mm) (%) 18 % Silt (0.063mm - 0.0312mm) (%) 13 % Silt (0.0312mm - 0.004mm) (%) 8 % Clay (<4um) (%) 6 Leachable Anions Total Nitrogen by LECO (%) 0.07 0.13 0.16 0.07 & Nutrients Organic / Total Organic Carbon (%) 0.8 2.4 1.8 0.7 Inorganic Carbon L742011 CONTD.... PAGE 8 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-31 L742011-32 L742011-33 L742011-34 L742011-35 Description Sampled Date 11-MAR-09 11-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 15:00 15:00 09:40 09:40 09:40 Client ID EEMCY5-CRB3-03 EEMCY5-CRB3-04 EEMCY5-CRB2A- EEMCY5-CRB2A- EEMCY5-CRB2A- 01 02 03
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) 4 % Sand (2.00mm - 1.00mm) (%) 3 % Sand (1.00mm - 0.50mm) (%) 7 % Sand (0.50mm - 0.25mm) (%) 20 % Sand (0.25mm - 0.125mm) (%) 28 % Sand (0.125mm - 0.063mm) (%) 15 % Silt (0.063mm - 0.0312mm) (%) 8 % Silt (0.0312mm - 0.004mm) (%) 9 % Clay (<4um) (%) 6 Leachable Anions Total Nitrogen by LECO (%) 0.09 0.08 0.09 0.09 & Nutrients Organic / Total Organic Carbon (%) 0.9 1.3 2.0 1.5 Inorganic Carbon L742011 CONTD.... PAGE 9 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-36 L742011-37 L742011-38 L742011-39 L742011-40 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 09:40 10:30 10:30 10:30 10:30 Client ID EEMCY5-CRB2A- EEMCY5-CRB6A- EEMCY5-CRB6A- EEMCY5-CRB6A- EEMCY5-CRB6A- 04 01 02 03 04
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) <1 1 % Sand (2.00mm - 1.00mm) (%) <1 4 % Sand (1.00mm - 0.50mm) (%) 3 12 % Sand (0.50mm - 0.25mm) (%) 10 40 % Sand (0.25mm - 0.125mm) (%) 46 29 % Sand (0.125mm - 0.063mm) (%) 24 4 % Silt (0.063mm - 0.0312mm) (%) 6 3 % Silt (0.0312mm - 0.004mm) (%) 6 4 % Clay (<4um) (%) 5 3 Leachable Anions Total Nitrogen by LECO (%) 0.06 0.08 0.07 & Nutrients Organic / Total Organic Carbon (%) 0.5 0.6 0.4 Inorganic Carbon L742011 CONTD.... PAGE 10 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-41 L742011-42 L742011-43 L742011-44 L742011-45 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 11:30 11:30 11:30 11:30 12:30 Client ID EEMCY5-CRB6B- EEMCY5-CRB6B- EEMCY5-CRB6B- EEMCY5-CRB6B- EEMCY5-CRB1A- 01 02 03 04 01
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) <1 % Sand (2.00mm - 1.00mm) (%) <1 % Sand (1.00mm - 0.50mm) (%) 2 % Sand (0.50mm - 0.25mm) (%) 26 % Sand (0.25mm - 0.125mm) (%) 57 % Sand (0.125mm - 0.063mm) (%) 7 % Silt (0.063mm - 0.0312mm) (%) 3 % Silt (0.0312mm - 0.004mm) (%) 2 % Clay (<4um) (%) 3 Leachable Anions Total Nitrogen by LECO (%) 0.04 0.04 0.04 0.13 & Nutrients Organic / Total Organic Carbon (%) 0.2 0.2 0.2 2.3 Inorganic Carbon L742011 CONTD.... PAGE 11 of 13 ALS LABORATORY GROUP ANALYTICAL REPORT 25-MAR-09 14:55
Sample ID L742011-46 L742011-47 L742011-48 Description Sampled Date 12-MAR-09 12-MAR-09 12-MAR-09 Sampled Time 12:30 12:30 12:30 Client ID EEMCY5-CRB1A- EEMCY5-CRB1A- EEMCY5-CRB1A- 02 03 04
Grouping Analyte SOIL Particle Size % Gravel (>2mm) (%) <1 % Sand (2.00mm - 1.00mm) (%) 1 % Sand (1.00mm - 0.50mm) (%) 3 % Sand (0.50mm - 0.25mm) (%) 12 % Sand (0.25mm - 0.125mm) (%) 36 % Sand (0.125mm - 0.063mm) (%) 22 % Silt (0.063mm - 0.0312mm) (%) 10 % Silt (0.0312mm - 0.004mm) (%) 10 % Clay (<4um) (%) 5 Leachable Anions Total Nitrogen by LECO (%) 0.11 0.12 & Nutrients Organic / Total Organic Carbon (%) 1.8 2.2 Inorganic Carbon L742011 CONTD.... PAGE 12 of 13 Reference Information 25-MAR-09 14:55
Additional Comments for Sample Listed: Samplenum Matrix Report Remarks Sample Comments
Methods Listed (if applicable): ALS Test Code Matrix Test Description Analytical Method Reference(Based On)
C-TOT-ORG-LECO-SK Soil Organic Carbon by combustion method SSSA (1996) p. 973 Total Organic Carbon (C-TOT-ORG-LECO-SK, C-TOT-ORG-SK)
Total C and inorganic C are determined on separate samples. The total C is determined by combustion and thermal conductivity detection, while inorganic C is determined by weight lass after addition of hydrochloric acid. Organic C is calculated by the difference between these two determinations.
Reference for Total C: Nelson, D.W. and Sommers, L.E. 1996. Total Carbon, organic carbon and organic matter. P. 961-1010 In: J.M. Bartels et al. (ed.) Methods of soil analysis: Part 3 Chemical methods. (3rd ed.) ASA and SSSA, Madison, WI. Book series no. 5
Reference for Inorganic C: Loeppert, R.H. and Suarez, D.L. 1996. Gravimetric Method for Loss of Carbon Dioxide. P. 455-456 In: J.M. Bartels et al. (ed.) Methods of soil analysis: Part 3 Chemical methods. (3rd ed.) ASA and SSSA, Madison, WI. Book series no. 5
N-TOT-LECO-SK Soil Total Nitrogen by combustion method SSSA (1996) p. 973-974 The sample is introduced into a quartz tube where it undergoes combustion at 900 C in the presence of oxygen. Combustion gases are first carried through a catalyst bed in the bottom of the combustion tube, where oxidation is completed and then carried through a reducing agent (copper), where the nitrogen oxides are reduced to elemental nitrogen. This mixture of N2, CO2, and H2O is then passed through an absorber column containing magnesium perchlorate to remove water. N2 and CO2 gases are then separated in a gas chromatographic column and detected by thermal conductivity.
Reference: Bremner, J.M. 1996. Nitrogen - Total (Dumas Methods). P. 1088 In: J.M. Bartels et al. (ed.) Methods of soil analysis: Part 3 Chemical methods. (3rd ed.) ASA and SSSA, Madison, WI. Book series no. 5
PSA-PIPET-DETAIL-SK Soil Particle size - Sieve and Pipette FORESTRY CANADA (1991) P. 46-48 MOD Particle size analysis involves the measurement of the proportions of the various primary soil particle sizes (ie. clay < 0.004 mm, silt 0.004-0.063 mm, sand 0.063-2.0 mm and gravel > 2.0 mm). In this method, the gravel and sand portions are determined by sieving, while the silt and clay portion is determined by sedimentation using Stokes Law, which relates the radius of the particles to the velocity of the sedimentation in water. Pretreatment of the soil with Calgon (sodium hexametaphosphate) is used to ensure the complete dispersion of the primary soil particles. Additional pretreatments may be necessary to remove cementing materials such as CaCO3 and organic matter.
Reference Y.P. Kalra, and D.G. Maynard, 1991. Methods Manual For Forest Soil and Plant Analysis,Northwest Region. Forestry Canada (modified sand, silt and clay size ranges)
** Laboratory Methods employed follow in-house procedures, which are generally based on nationally or internationally accepted methodologies. The last two letters of the above ALS Test Code column indicate the laboratory that performed analytical analysis for that test. Refer to the list below:
Laboratory Definition Code Laboratory Location Laboratory Definition Code Laboratory Location
SK ALS LABORATORY GROUP - SASKATOON, SASKATCHEWAN, CANADA L742011 CONTD.... PAGE 13 of 13 Reference Information
Methods Listed (if applicable): ALS Test Code Matrix Test Description Analytical Method Reference(Based On)
GLOSSARY OF REPORT TERMS Surr - A surrogate is an organic compound that is similar to the target analyte(s) in chemical composition and behavior but not normally detected in enviromental samples. Prior to sample processing, samples are fortified with one or more surrogate compounds. The reported surrogate recovery value provides a measure of method efficiency. mg/kg (units) - unit of concentration based on mass, parts per million mg/L (units) - unit of concentration based on volume, parts per million N/A - Result not available. Refer to qualifier code and definition for explanation
Test results reported relate only to the samples as received by the laboratory. UNLESS OTHERWISE STATED, ALL SAMPLES WERE RECEIVED IN ACCEPTABLE CONDITION. Although test results are generated under strict QA/QC protocols, any unsigned test reports, faxes, or emails are considered preliminary.
ALS Laboratory Group has an extensive QA/QC program where all analytical data reported is analyzed using approved referenced procedures followed by checks and reviews by senior managers and quality assurance personnel. However, since the results are obtained from chemical measurements and thus cannot be guaranteed, ALS Laboratory Group assumes no liability for the use or interpretation of the results.
Appendix A6
Benthic Survey: Aquametrix Analytical Reports
Temperature Corrected Redox Station Replicate Sulphide (μM) Redox (mV) (°C) (mV) 1 85.8 -74.5 6.4 143.1 CRB7 2 86.2 -42.6 5.1 176.3 3 77.4 -71.2 5.7 147.1 1 86.2 -93.0 6.9 124.1 CRB6 2 84.2 -136.5 6.2 81.3 3 15.1 -162.2 6.0 55.8 1 42.8 -183.8 5.9 34.3 CRB5C 2 44.4 -132.1 5.4 86.5 3 29.4 -201.4 7.7 14.9 1 79.3 -191.0 8.6 24.4 CRB5A 2 66.3 -180.5 8.3 35.2 3 85.8 -190.7 7.8 25.5 1 81.4 -185.5 10.8 28.3 CRB2 2 50.2 -175.7 10.2 38.1 3 41.6 -238.9 10.3 -25.2 1 20.3 -62.4 12.7 148.9 CRB5B 2 27.1 -154.6 11.8 57.6 3 3.81 -80.3 13.2 130.5 1 74.2 -208.1 10.1 5.8 CRB4A 2 28.6 -156.1 13.5 54.4 3 44.0 -165.5 13.8 44.7 1 12.1 -106.3 12.9 104.8 CRB3 2 9.76 -86.7 11.8 125.5 3 14.3 -105.5 11.9 106.6 1 69.0 -243.4 8.6 -28.0 CRB2A 2 87.8 -195.1 8.2 20.7 3 69.0 -203.1 8.8 12.1 1 23.0 -146.4 8.4 69.2 CRB6A 2 68.1 -224.6 7.8 -8.4 3 19.3 -193.5 8.6 21.9 1 23.6 -161.3 8.6 54.1 CRB6B 2 25.3 -134.6 9.3 80.1 3 12.9 -131.9 8.1 84.0 1 59.7 -215.2 8.9 -0.1 CRB1A 2 56.9 -234.6 9.7 -20.3 3 62.2 -231.7 10.3 -18.0
Appendix A7
Benthic Survey: Columbia Science Invertebrate Data and QA/QC Reports
Hatfield Consultants - Crofton 2009 Prepared by Columbia Science
SUMMARY OF BENTHIC INVERTEBRATE SAMPLES -CROFTON MARCH 9-12, 2009
Substrate Substrate Station Volume Characteristics (ml)
CRB 1A-1 2200 Coarse sand and fine woody debris CRB 1A-2 1500 Coarse sand and fine woody debris CRB 1A-3 1500 Coarse sand and fine woody debris CRB 2-1 1300 Woody fibre and coarse sand CRB 2-2 1200 Woody fibre and coarse sand CRB 2-3 1300 Woody fibre and coarse sand CRB 2A-1 600 Coarse sand and fine woody debris CRB 2A-2 500 Coarse sand and fine woody debris CRB 2A-3 1000 Coarse sand and fine woody debris CRB 3-1` 5400 Coarse sand, gravel, fine woody debris CRB 3-2 4300 Coarse sand, gravel, fine woody debris CRB 3-3 1800 Coarse sand, gravel, fine woody debris CRB 4A-1* 2200 Woody fibre and coarse sand CRB 4A-2 1300 Woody fibre and coarse sand CRB 4A-3** 3300 Woody fibre and coarse sand CRB 5A-1 4900 Coarse sand, gravel, fine woody debris CRB 5A-2 2000 Coarse sand, gravel, fine woody debris CRB 5A-3 4100 Coarse sand, gravel, fine woody debris CRB 5B-1 300 Coarse sand, gravel, fine woody debris CRB 5B-2 1900 Coarse sand, gravel, fine woody debris CRB 5B-3 2550 Coarse sand, gravel, fine woody debris CRB 5C-1 300 Mud CRB 5C-2 200 Mud CRB 5C-3 300 Mud CRB 6-1 6200 Coarse sand, gravel, fine woody debris CRB 6-2 7100 Coarse sand, gravel, fine woody debris CRB 6-3 3300 Coarse sand, gravel, fine woody debris CRB 6A-1 1800 Coarse sand and fine woody debris CRB 6A-2 1300 Coarse sand and fine woody debris CRB 6A-3 3000 Coarse sand and fine woody debris CRB 6B-1 500 Coarse sand CRB 6B-2 700 Coarse sand CRB 6B-3 300 Coarse sand CRB 7-1 200 Coarse sand CRB 7-2 400 Coarse sand CRB 7-3 400 Coarse sand Prepared by COLUMBIA SCIENCE Prepared for Hatfield Consultants, Ltd. DATA REPORT CROFTON 2009
Submitting Firm Columbia Science Submitter's Name Sandy Lipovsky Address 275 Inverness Rd. Courtenay, BC V9N 9S6 Phone/Fax 250-335-0714 Email [email protected] Sample State Description BI Level of Lowest Sample Preservation 10% formalin/70%Condition of Organisms Good Dates Analyzed April - November 2009 Taxonomist Sandy Lipovsky
Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad
CNIDARIA Hydrozoa Campanulariidae colony frag. Corymorphidae Indet. Monobrachium parasitum colony frag. Perigonimus repens colony fragment 1 Anthozoa Halcampa decemtentaculata 11 Halipteris californica Metridium farcimen Pachycerianthus fimbriatus Pennatulacea Indet. 211 Virgularia sp. 21 PLATYHELMINTHES Stylochidae Indet. 1
Page 1 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad NEMERTEA Anopla Indet. Carinoma mutabilis 2 Cerebratulus californiensis 1 1 1 1 1 115332 Hoplonemertea Indet. 1 Lineidae 32 Lineus sp. Palaeonemertea Paranemertes californica 1 1 3 Tubulanidae 1 Tubulanus nothus 542314393 Tubulanus polymorphus 9 1 5 4 1 12 11 21439 ANNELIDA Polychaeta Errantia Arctonoe sp. Dorvillea longicornis Dorvillea sp. Driloneris longa Errano bicirrata 1 Eteone californica Eteone longa 2 4 3 4 2 2 Eteone sp. Eteone spilotus Eulalia californiensis Eulalia sp. 11 Eumida longicornuta Eusyllis sp. Exogone spp. Glycera americana 11 Glycera nana 62145763595675798 Glycera robusta Glycinde armigera 3 112334
Page 2 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Goniada brunnea 1211 Harmothoe imbricata Cmplx. 2 1 1 Hesionidae Indet. Lepidasthenia berkeleyae 1 3 1 1 Lumbrineridae Indet. 5 8 5 11 15 9 Lumbrineris californiensis 1 Lumbrineris cruzensis 3 5 5 1 8 15 18 20 1 18 21 Lumbrineris japonica Lumbrineris latreilli 4 Lumbrineris spp. 222 7 Malmgreniella macginitiei Malmgreniella nigralba Nephtys cornuta 1433611 244 1 8 Nephtys ferruginea 223112 22 2 Nereis procera 77645855437343432 Odontosyllis phosphorea Parandalia fauveli 13 Pholoe glabra 232 14 1 Pholoides asperus 12 Phyllodoce groenlandica 2 1 1 Phyllodoce hartmanae 1 1 1 Phyllodoce sp. 1 Pilargis berkeleyae 22312221214 Podarkeopsis glabrus 6 3 4 3 1 3 10 1 Polynoidae Indet. 1 Scoletoma luti 3 2 3 1 348545 Sigambra bassi 431672 1 Sige sp. Sphaerodoropsis sphaerulifer Sthenelais berkeleyae Syllis elongata 1 Syllis sp.
Page 3 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Tenonia priops 1 Typosyllis alternata 2 Polychaeta Sedentaria Amaena occidentalis 1 Ampharete finmarchica 1 Ampharete labrops Ampharete sp. Ampharetidae Indet. Aphelochaeta glandaria 1 1121 Aphelochaeta monilaris 111 Aphelochaeta sp. 11 Aricidea lopezi 4 2 2311 1 Armandia brevis 12 Artacama coniferi Barantolla americana Boccardiella hamata Capitella capitata Cmplx. Chaetozone setosa 2 1 4 3 3 3 2 Chaetozone sp. 11 Chone sp. Cirratulidae Indet. Cossura pygodactylata 21 4 1 16 2 10 7 6 Decamastus gracilis 146 140 159 122 186 230 129 18 96 21 150 Dipolydora cardalia Dipolydora quadrilobata 225 Dipolydora socialis 84 79 138 12 63 52 110 138 190 Dipolydora sp. Euchone analis Euclymene sp. 1 Euclymeninae Indet. 3 1 Galathowenia oculata 34 59 79 109 86 111 30 45 56 Heteromastus filiformis 3 1
Page 4 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Heteromastus filobranchus 3 1 2 2 3 Heteromastus spp. 1 1 4 Lanassa gracilis Laonice cirrata 37435136 4 6 4 1 Leitoscoloplos pugettensis 8 2 7 4 1 1 1 Levinsenia gracilis 26 14 10 4 15 10 11 6 22 Lumbriclymeninae Indet. 5 9 7 14 3 Magelona longicornis 12 Maldane sarsi Mediomastus spp. 291 202 301 174 442 335 206 108 531 Megalomma splendida Metasychis disparidentata Mugga wahrbergi Neosabellaria cementarium Nichomache lumbricalis Nichomache personata Notomastus hemipodus 5 10 11 11 7 14 Notomastus sp. 3 Notomastus tenuis Notoproctus pacificus Onuphis iridescens 111122124211 Ophelina acuminata 1 1 6 2 Owenia fusiformis 1 Paraprionospio alata 2 2 1312322 2 5 Pectinaria californiensis 5 1 4 5337768 1 Pectinaria granulata Petaloproctus borealis Pherusa sp. 121 Phylo felix Pista agassizi Pista brevibranchiata Pista sp.
Page 5 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Pista wui Polycirrus sp. 4 1111 1 1 Polydora spp. 2 3 9 Polydora websteri 2 Praxillella gracilis 1 Praxillella pacifica 4 Praxillella praetermissa 2 4 1 Praxillella sp. 1 2 Prionospio (Minuspio) lighti 29 5 14 4 17 14 2 2 3 Pionospio (Minuspio) multibranchiata 1 2 2 1 1 Prionospio dubia 2125 42 Prionospio jubata 62133285133 Pseudopolydora kempi 53 Rhodine bitorquata Rhynchospio glutaea 1 Sabellidae Indet. Scalibregma californicum 3437 91211 Scionella japonica Scoloplos acmeceps 2 6 1 4 64434 Spio cirrifera 222 Spiochaetopterus costarum 111 Spiophanes berkeleyorum 2 3 2 2 1 1 1 4 3 1 4 Spiophanes fimbriata 7 3 3 3 1 Spiophanes sp. Sternaspis fossor 1 1 2 1 Streblosoma pacifica Terebellidae Indet. 2 Terebellides californica 2 1 7 1 2 Terebellides sp. 2 2 5 2 Thelepus sp. Trochochaeta multisetosa Oligochaeta
Page 6 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Tubificidae 2 2 6 1 33129 SPIUNCULA Nephasoma diaphanes Thysanocardia nigra 1 1 1 1 1 MOLLUSCA Aplacophora Chaetoderma argenteum 3 Chaetoderma elegans Chaetoderma sp. 11 Polyplacophora Ischnochitonidae Indet. Gastropoda Alvania rosana Astyris gausapata 6 2 1 16 2 9 1 1 1 Corambe sp. Cylichna attonsa 1 1 2 1 1 Euspira sp. Gastropoda Indet. (dissolved) 2 Lottidae Indet. Nassarius mendicus Odostomia sp. Turbonilla sp. Turridae Indet. Bivalvia Adontorhina cyclia 1 2 5 2 Axinopsida serricata 411931543410 6 8 Cardiomya pectinata 1 Chlamys sp. Compsomyax subdiaphana 1 31 Crenella decussata Cyclocardia ventricosa Delectopecten vancouverensis
Page 7 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Ennucula tenuis Lasaeidae Indet. 1 Lucinoma annulatum 61514 1 16861 Lyonsia californica 1 Macoma carlottensis 7592438121 Macoma elimata 2 4 14 3 4 17 3 1 Macoma golikovi 1 Macoma spp. 11 12 4 6 1 4 Macoma yoldiformis Megayoldia thracaeiformis Mya sp. Mya truncata Naeromya sp. Nemocardium centifilosum 1 1 1 1 1 2 Nutricola lordi Parvilucina tenuisculpta 54 31 54 19 75 39 34 14 56 23 45 20 56 14 51 3 63 Rochefortia tumida 11 Solamen columbianum Tellina sp. Thyasira flexuosa Yoldia seminuda Yoldia sp. Scaphopoda Antalis pretiosum Pulsellum salishorum ARTHROPODA Amphipoda Americhelidium shoemakeri 1 Ampelisca lobata Ampelisca sp. 113 1 Aoroides columbiae 21 1 Aoroides sp. 1
Page 8 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Bathymedon pumilis Bruzelia tuberculata 1 3 2 Byblis millsi Desdimelita desdichada Dyopedos arcticus 1 Eobrolgus chumashi 1 Guernea reduncans Heterophoxus affinis 1 4 31332 Heterophoxus conlonae Heterophoxus ellisi Leucothoidae Indet. Melita oregonensis Monoculodes perditus Oedicerotidae Indet. Opisa tridentata 1111121 Orchomene sp. 1 Pachynus barnardi 1 1 Parametaphoxus quayleyi 11 Photis sp. Pleustidae Indet. 1 Pleusymtes subglaber 1 2 Protomedeia sp. Rhachotropis barnardi Rhepoxynius tridentatus Westwoodilla tone 23146 1 Copepoda Harpacticoida Tanaidacea Leptochelia savignyi 1 Scolaura phillipsi 21 Ostracoda Cylindroleberidae 245
Page 9 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Euphilomedes carcharodonta 1 2 1 2 2 Euphilomedes producta 31 25 22 9 27 8 11 21 8 12 Rutiderma lomae Scleroconcha trituberculata Cumacea Campylaspis canaliculata Campylaspis sp. Diastylis santamariensis 2 Diastylis sp. 11 Eudorella pacifica 1 Eudorellopsis longirostris Leucon sp. 1 Leucon subnasica Decapoda Crangonidae Hemigrapsus sp. Oregonia gracilis Pinnixa sp. 3 3 1 PHORONIDA Phoronis muelleri 2 BRYOZOA Bowerbankia gracilis colony frag. Tubuliporidae colony frag. ECHINODERMATA Ophiuroidea Amphiodia urtica 111 Ophiura sarsi Ophiura sp. Holothuroidea Pentamera populifera 11112111451 Pentamera pseudocalcigera Pentamera rigida
Page 10 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB 1A CRB2 CRB2 CRB2 CRB2 CRB2 CRB2 CRB2A CRB2A CRB2A CRB2A CRB2A Sampling Date (m/d/y) 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/09/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 Replicate 1 1223311223311223 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad HEMICHORDATA Saccoglossus sp. 2 2 4 1 1 1
Notes: Calanoid Gastropod egg cases Nematoda 1 2 Leptomedusae (clear jellyfish) 1 1
Page 11 Crofton 2009 Benthic Invertebrate Data Prepared by COLUMBIA SCIENCE Prepared for Hatfield Consultants, Ltd. DATA REPORT CROFTON 2009
Submitting Firm Submitter's Name Address Phone/Fax Sample State Description Sample Preservation Dates Analyzed Taxonomist
Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv
CNIDARIA Hydrozoa Campanulariidae colony frag. Corymorphidae Indet. 1 Monobrachium parasitum colony frag. Perigonimus repens colony fragment 11 1 Anthozoa Halcampa decemtentaculata Halipteris californica Metridium farcimen Pachycerianthus fimbriatus Pennatulacea Indet. 121 21 Virgularia sp. PLATYHELMINTHES Stylochidae Indet. 2
Page 12 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv NEMERTEA Anopla Indet. Carinoma mutabilis 11 Cerebratulus californiensis 21 11 32 2104142 Hoplonemertea Indet. Lineidae 1215 Lineus sp. Palaeonemertea Paranemertes californica Tubulanidae Tubulanus nothus 363 17 Tubulanus polymorphus 45 9 10 4 3 2 ANNELIDA Polychaeta Errantia Arctonoe sp. Dorvillea longicornis Dorvillea sp. 1 Driloneris longa 1 4 Errano bicirrata 21 1 Eteone californica 11 Eteone longa 2 Eteone sp. 484 Eteone spilotus 11412 Eulalia californiensis Eulalia sp. Eumida longicornuta 1 Eusyllis sp. Exogone spp. 2 Glycera americana 1 Glycera nana 75797352 21 81727 Glycera robusta Glycinde armigera 11 139423
Page 13 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Goniada brunnea 4 Harmothoe imbricata Cmplx. 1 Hesionidae Indet. 1 Lepidasthenia berkeleyae Lumbrineridae Indet. Lumbrineris californiensis Lumbrineris cruzensis 22 21 17 15 14 28 9 11 Lumbrineris japonica 43 Lumbrineris latreilli 4 512 Lumbrineris spp. 16 10 3 3 12 Malmgreniella macginitiei 11111 Malmgreniella nigralba Nephtys cornuta 51 2524 Nephtys ferruginea 21 12 3 1 1 Nereis procera 31 6 2 11 412 Odontosyllis phosphorea Parandalia fauveli 241 12 Pholoe glabra 12 1 Pholoides asperus 4567 11 2 2 Phyllodoce groenlandica 1124 Phyllodoce hartmanae Phyllodoce sp. Pilargis berkeleyae 111427 1 4 Podarkeopsis glabrus 1 1 14 20 20 Polynoidae Indet. Scoletoma luti 8101261377 Sigambra bassi 1 2 36 10 41 1 36 12 Sige sp. 1 Sphaerodoropsis sphaerulifer 1 Sthenelais berkeleyae 1 Syllis elongata 34 Syllis sp. 22
Page 14 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Tenonia priops Typosyllis alternata 113 1 4 212 Polychaeta Sedentaria Amaena occidentalis 2 Ampharete finmarchica 13 Ampharete labrops 2 Ampharete sp. 1 Ampharetidae Indet. 1 Aphelochaeta glandaria 51 12 Aphelochaeta monilaris 142345 Aphelochaeta sp. Aricidea lopezi 1 12 Armandia brevis 224 Artacama coniferi 1 Barantolla americana 2 Boccardiella hamata Capitella capitata Cmplx. 24 Chaetozone setosa 1 1 Chaetozone sp. 2 Chone sp. 14 Cirratulidae Indet. Cossura pygodactylata 1 Decamastus gracilis 19 58 70 108 20 8 34 Dipolydora cardalia Dipolydora quadrilobata Dipolydora socialis 2 17 17 174 114 248 1 1 Dipolydora sp. Euchone analis Euclymene sp. Euclymeninae Indet. 12 1 Galathowenia oculata 448 232 250 229 204 Heteromastus filiformis
Page 15 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Heteromastus filobranchus 68 155 115 Heteromastus spp. 1 Lanassa gracilis Laonice cirrata 1111 Leitoscoloplos pugettensis 315 16 Levinsenia gracilis 547 1 Lumbriclymeninae Indet. 4 Magelona longicornis 2131135 Maldane sarsi 2 Mediomastus spp. 47 51 102 16 26 27 24 10 Megalomma splendida 313 1 Metasychis disparidentata 3 1 Mugga wahrbergi 2 Neosabellaria cementarium 21 Nichomache lumbricalis 1 Nichomache personata Notomastus hemipodus 5153 Notomastus sp. Notomastus tenuis 68 11 Notoproctus pacificus 121 52 Onuphis iridescens 42 62 2 4 81 Ophelina acuminata 4 Owenia fusiformis 125 Paraprionospio alata 21 2 1761039 2 Pectinaria californiensis 5531863111261045132 Pectinaria granulata 112 Petaloproctus borealis 1 Pherusa sp. 1 Phylo felix Pista agassizi 3 Pista brevibranchiata Pista sp. 12 3
Page 16 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Pista wui 1 Polycirrus sp. 511151 11 Polydora spp. Polydora websteri Praxillella gracilis Praxillella pacifica 1 Praxillella praetermissa 11 Praxillella sp. 1 Prionospio (Minuspio) lighti 1 4 2 10 11 12 Pionospio (Minuspio) multibranchiata 422 Prionospio dubia 444 4 Prionospio jubata 41 72122 1428 2 Pseudopolydora kempi 2 Rhodine bitorquata Rhynchospio glutaea Sabellidae Indet. Scalibregma californicum 2222314 Scionella japonica 1 Scoloplos acmeceps 132121 Spio cirrifera 11221 Spiochaetopterus costarum 431 11 Spiophanes berkeleyorum 1431 4 1 Spiophanes fimbriata 3722 Spiophanes sp. Sternaspis fossor 13 12 2 4 Streblosoma pacifica Terebellidae Indet. Terebellides californica 11 1 Terebellides sp. Thelepus sp. Trochochaeta multisetosa 1 11 Oligochaeta
Page 17 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Tubificidae SPIUNCULA Nephasoma diaphanes 1 Thysanocardia nigra 11611 MOLLUSCA Aplacophora Chaetoderma argenteum Chaetoderma elegans 21 113 Chaetoderma sp. Polyplacophora Ischnochitonidae Indet. 3 Gastropoda Alvania rosana Astyris gausapata 22644 105 Corambe sp. Cylichna attonsa 152 Euspira sp. 1 Gastropoda Indet. (dissolved) Lottidae Indet. 21 Nassarius mendicus 1 Odostomia sp. Turbonilla sp. 14 2 Turridae Indet. Bivalvia Adontorhina cyclia Axinopsida serricata 756657 7263 Cardiomya pectinata 21 1 Chlamys sp. Compsomyax subdiaphana 52 11 2 1 Crenella decussata Cyclocardia ventricosa Delectopecten vancouverensis 11
Page 18 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ennucula tenuis Lasaeidae Indet. Lucinoma annulatum 1 1029 1 2 2163 Lyonsia californica Macoma carlottensis 3 4 6 2 120 82 91 40 120 113 3 9 Macoma elimata 12229 241 Macoma golikovi 11 Macoma spp. 5 3 10 12 1 2 Macoma yoldiformis 3 Megayoldia thracaeiformis Mya sp. Mya truncata Naeromya sp. Nemocardium centifilosum 1 313 Nutricola lordi Parvilucina tenuisculpta 19 44 7 48 21 57 14 266 28 490 78 267 38 23 1 61 10 Rochefortia tumida 131 Solamen columbianum 3 11 Tellina sp. Thyasira flexuosa 1 1 Yoldia seminuda 2 Yoldia sp. 4 Scaphopoda Antalis pretiosum 1 Pulsellum salishorum ARTHROPODA Amphipoda Americhelidium shoemakeri 2 Ampelisca lobata 11 Ampelisca sp. 12 1 Aoroides columbiae 9 Aoroides sp.
Page 19 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Bathymedon pumilis Bruzelia tuberculata Byblis millsi 11 22 Desdimelita desdichada Dyopedos arcticus Eobrolgus chumashi Guernea reduncans Heterophoxus affinis 883 Heterophoxus conlonae Heterophoxus ellisi 121 Leucothoidae Indet. Melita oregonensis Monoculodes perditus Oedicerotidae Indet. Opisa tridentata Orchomene sp. 2 Pachynus barnardi Parametaphoxus quayleyi 17 1 Photis sp. Pleustidae Indet. Pleusymtes subglaber Protomedeia sp. Rhachotropis barnardi 1 Rhepoxynius tridentatus 1 Westwoodilla tone 12 22 7 Copepoda Harpacticoida Tanaidacea Leptochelia savignyi 228 Scolaura phillipsi 411 4 11 Ostracoda Cylindroleberidae 1
Page 20 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Euphilomedes carcharodonta 4 6284 Euphilomedes producta 35 29 30 2 5 18 Rutiderma lomae 6 Scleroconcha trituberculata 1 Cumacea Campylaspis canaliculata Campylaspis sp. Diastylis santamariensis Diastylis sp. 2 Eudorella pacifica 16 1 3 Eudorellopsis longirostris Leucon sp. 1 Leucon subnasica Decapoda Crangonidae Hemigrapsus sp. 1 Oregonia gracilis Pinnixa sp. 33 PHORONIDA Phoronis muelleri 31 17 7 4 9 4 BRYOZOA Bowerbankia gracilis colony frag. 11 Tubuliporidae colony frag. ECHINODERMATA Ophiuroidea Amphiodia urtica 3 5 5111 Ophiura sarsi 2 Ophiura sp. 1 Holothuroidea Pentamera populifera 22124 2 Pentamera pseudocalcigera Pentamera rigida 11
Page 21 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB2A CRB3 CRB3 CRB3 CRB3 CRB3 CRB3 CRB4A CRB4A CRB4A CRB4A CRB4A CRB4A CRB5A CRB5A CRB5A CRB5A Sampling Date (m/d/y) 3/12/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/11/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 31122331122331122 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv HEMICHORDATA Saccoglossus sp.
Notes: Calanoid Gastropod egg cases Nematoda Leptomedusae (clear jellyfish)
Page 22 Crofton 2009 Benthic Invertebrate Data Prepared by COLUMBIA SCIENCE Prepared for Hatfield Consultants, Ltd. DATA REPORT CROFTON 2009
Submitting Firm Submitter's Name Address Phone/Fax Sample State Description Sample Preservation Dates Analyzed Taxonomist
Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad
CNIDARIA Hydrozoa Campanulariidae colony frag. 11 Corymorphidae Indet. Monobrachium parasitum colony frag. Perigonimus repens colony fragment Anthozoa Halcampa decemtentaculata Halipteris californica 21 Metridium farcimen 2 Pachycerianthus fimbriatus 211 2 Pennatulacea Indet. 11 5 Virgularia sp. PLATYHELMINTHES Stylochidae Indet.
Page 23 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad NEMERTEA Anopla Indet. Carinoma mutabilis 1 Cerebratulus californiensis 221126 1 728 Hoplonemertea Indet. Lineidae 1 2 Lineus sp. 3 Palaeonemertea 1 Paranemertes californica Tubulanidae Tubulanus nothus 4126 3 Tubulanus polymorphus 1211 62 ANNELIDA Polychaeta Errantia Arctonoe sp. 1 Dorvillea longicornis Dorvillea sp. Driloneris longa 2 2 Errano bicirrata 1 12 Eteone californica Eteone longa 1 Eteone sp. Eteone spilotus 12 Eulalia californiensis 10 1 Eulalia sp. Eumida longicornuta Eusyllis sp. 1 Exogone spp. 11 Glycera americana Glycera nana 6231036433 2331553 Glycera robusta 1 Glycinde armigera 1121 124
Page 24 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Goniada brunnea 21 Harmothoe imbricata Cmplx. 1 Hesionidae Indet. Lepidasthenia berkeleyae Lumbrineridae Indet. Lumbrineris californiensis 15 1 Lumbrineris cruzensis 6 9 14 9 1 33 11 Lumbrineris japonica Lumbrineris latreilli 2 Lumbrineris spp. 4 1 Malmgreniella macginitiei 111221 Malmgreniella nigralba 1 Nephtys cornuta 111 Nephtys ferruginea 1122132 Nereis procera 5221 122 1 Odontosyllis phosphorea Parandalia fauveli 231 Pholoe glabra 1 Pholoides asperus 31 13 1 1 38 14 Phyllodoce groenlandica Phyllodoce hartmanae Phyllodoce sp. Pilargis berkeleyae 1 111 Podarkeopsis glabrus 1114 Polynoidae Indet. Scoletoma luti 2133 2736 Sigambra bassi 22 11 2 3 Sige sp. Sphaerodoropsis sphaerulifer 1 Sthenelais berkeleyae 132 Syllis elongata 131523 16 Syllis sp.
Page 25 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Tenonia priops 1 Typosyllis alternata 124 1 Polychaeta Sedentaria Amaena occidentalis Ampharete finmarchica 1 Ampharete labrops Ampharete sp. Ampharetidae Indet. 1 Aphelochaeta glandaria 2111 25 Aphelochaeta monilaris 721 21 6 Aphelochaeta sp. Aricidea lopezi 112 Armandia brevis 1 Artacama coniferi 1 Barantolla americana 1 Boccardiella hamata 6 Capitella capitata Cmplx. 1 Chaetozone setosa 3 Chaetozone sp. 1 Chone sp. 1 Cirratulidae Indet. Cossura pygodactylata Decamastus gracilis 9534528 8270 Dipolydora cardalia 51 Dipolydora quadrilobata 1 Dipolydora socialis 52 6 15 4 1 Dipolydora sp. 11 1 Euchone analis Euclymene sp. 1 Euclymeninae Indet. 23 Galathowenia oculata 256 152 173 198 81 75 Heteromastus filiformis
Page 26 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Heteromastus filobranchus 1 72 32 101 Heteromastus spp. 478 Lanassa gracilis Laonice cirrata 11 Leitoscoloplos pugettensis 2 Levinsenia gracilis 22 Lumbriclymeninae Indet. Magelona longicornis 3152 7 9 11 Maldane sarsi Mediomastus spp. 4 43 33 36 8 10 11 14 4 Megalomma splendida 21 4 Metasychis disparidentata 1 Mugga wahrbergi Neosabellaria cementarium Nichomache lumbricalis 314 272 Nichomache personata 1 Notomastus hemipodus 18 2 11 18 Notomastus sp. Notomastus tenuis 21 13 7 Notoproctus pacificus 2 4 Onuphis iridescens 2 11226 7 3 Ophelina acuminata 1 Owenia fusiformis Paraprionospio alata 1 111 2111 221 Pectinaria californiensis 1132 1 1 Pectinaria granulata 1 1 Petaloproctus borealis 22 1 51 Pherusa sp. 11 Phylo felix 1 1 Pista agassizi Pista brevibranchiata 1 3 Pista sp.
Page 27 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Pista wui 1 Polycirrus sp. 215 1 4 Polydora spp. 1 Polydora websteri Praxillella gracilis Praxillella pacifica 321 Praxillella praetermissa Praxillella sp. Prionospio (Minuspio) lighti 110 Pionospio (Minuspio) multibranchiata 41 Prionospio dubia Prionospio jubata 11 4 Pseudopolydora kempi 1 Rhodine bitorquata 43 Rhynchospio glutaea Sabellidae Indet. 1 Scalibregma californicum 143 67 Scionella japonica 4 Scoloplos acmeceps 211 2 Spio cirrifera 12 3 1111 Spiochaetopterus costarum 4412 Spiophanes berkeleyorum 12 54 1 1 5 Spiophanes fimbriata 2 Spiophanes sp. 1 Sternaspis fossor 1 Streblosoma pacifica Terebellidae Indet. Terebellides californica 12 Terebellides sp. 1 Thelepus sp. Trochochaeta multisetosa 1222 Oligochaeta
Page 28 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Tubificidae 2 SPIUNCULA Nephasoma diaphanes 8 Thysanocardia nigra 511 12 MOLLUSCA Aplacophora Chaetoderma argenteum Chaetoderma elegans Chaetoderma sp. Polyplacophora Ischnochitonidae Indet. Gastropoda Alvania rosana 1 Astyris gausapata 311 1 Corambe sp. Cylichna attonsa 11 5 1 Euspira sp. Gastropoda Indet. (dissolved) 42 Lottidae Indet. Nassarius mendicus 1 Odostomia sp. Turbonilla sp. 2 Turridae Indet. Bivalvia Adontorhina cyclia 2 Axinopsida serricata 36 1 12 9 13 3 22 21 52 6 17 Cardiomya pectinata 11 Chlamys sp. 1 Compsomyax subdiaphana 1 12 121112 Crenella decussata Cyclocardia ventricosa 1 Delectopecten vancouverensis
Page 29 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Ennucula tenuis 2 Lasaeidae Indet. Lucinoma annulatum 12 3 12 6 2 2 1 1 Lyonsia californica 1 Macoma carlottensis 16 10 3 21 34 4 16 1 41 13 Macoma elimata 91310 47 261 Macoma golikovi Macoma spp. 11 17 18 7 1 Macoma yoldiformis 1 Megayoldia thracaeiformis 1 Mya sp. Mya truncata 1 Naeromya sp. 11 Nemocardium centifilosum 31 2 1 Nutricola lordi 1 Parvilucina tenuisculpta 32 4 90 14 63 13 50 20 106 30 95 19 86 53 22 18 Rochefortia tumida 2513 Solamen columbianum 21 Tellina sp. Thyasira flexuosa Yoldia seminuda Yoldia sp. Scaphopoda Antalis pretiosum Pulsellum salishorum ARTHROPODA Amphipoda Americhelidium shoemakeri Ampelisca lobata 1 Ampelisca sp. 13 Aoroides columbiae Aoroides sp. 4
Page 30 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Bathymedon pumilis 1 Bruzelia tuberculata Byblis millsi 1 11 Desdimelita desdichada 1 1 Dyopedos arcticus Eobrolgus chumashi Guernea reduncans 1 2 Heterophoxus affinis 1 Heterophoxus conlonae 2 Heterophoxus ellisi 72 1 3 9 Leucothoidae Indet. 1 Melita oregonensis Monoculodes perditus 12 Oedicerotidae Indet. Opisa tridentata Orchomene sp. Pachynus barnardi 1 Parametaphoxus quayleyi 2 4 Photis sp. 2 Pleustidae Indet. Pleusymtes subglaber Protomedeia sp. Rhachotropis barnardi 11 2 Rhepoxynius tridentatus Westwoodilla tone 11 21 Copepoda Harpacticoida 1 Tanaidacea Leptochelia savignyi Scolaura phillipsi 1112 21 Ostracoda Cylindroleberidae
Page 31 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Euphilomedes carcharodonta 23 Euphilomedes producta 8 1 10 5 9 34 19 Rutiderma lomae 1 20 3 Scleroconcha trituberculata Cumacea Campylaspis canaliculata 1 Campylaspis sp. Diastylis santamariensis 11 Diastylis sp. 1 Eudorella pacifica 11 Eudorellopsis longirostris 31 Leucon sp. Leucon subnasica Decapoda Crangonidae Hemigrapsus sp. Oregonia gracilis Pinnixa sp. 1 1111 PHORONIDA Phoronis muelleri 9258 17 BRYOZOA Bowerbankia gracilis colony frag. 11 Tubuliporidae colony frag. ECHINODERMATA Ophiuroidea Amphiodia urtica 1221 Ophiura sarsi Ophiura sp. Holothuroidea Pentamera populifera 21 11 Pentamera pseudocalcigera Pentamera rigida 1
Page 32 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB5A CRB5A CRB5B CRB5B CRB5B CRB5B CRB5B CRB5B CRB5C CRB5C CRB5C CRB5C CRB5C CRB5C CRB6 CRB6 CRB6 Sampling Date (m/d/y) 3/10/09 3/10/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/9/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 33112233112233112 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad HEMICHORDATA Saccoglossus sp. 1
Notes: Calanoid Gastropod egg cases 2 Nematoda 1 Leptomedusae (clear jellyfish)
Page 33 Crofton 2009 Benthic Invertebrate Data Prepared by COLUMBIA SCIENCE Prepared for Hatfield Consultants, Ltd. DATA REPORT CROFTON 2009
Submitting Firm Submitter's Name Address Phone/Fax Sample State Description Sample Preservation Dates Analyzed Taxonomist
Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad
CNIDARIA Hydrozoa Campanulariidae colony frag. Corymorphidae Indet. Monobrachium parasitum colony frag. 1 Perigonimus repens colony fragment 1 Anthozoa Halcampa decemtentaculata 2 Halipteris californica 1 Metridium farcimen Pachycerianthus fimbriatus 2 Pennatulacea Indet. 11 1 Virgularia sp. PLATYHELMINTHES Stylochidae Indet.
Page 34 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad NEMERTEA Anopla Indet. 1 Carinoma mutabilis 1 Cerebratulus californiensis 451 1 1 1 222 21 Hoplonemertea Indet. Lineidae 22 2 Lineus sp. Palaeonemertea Paranemertes californica Tubulanidae Tubulanus nothus 1113 5 Tubulanus polymorphus 334 6 413 ANNELIDA Polychaeta Errantia Arctonoe sp. Dorvillea longicornis 2 Dorvillea sp. 1 11 Driloneris longa 12 2 Errano bicirrata Eteone californica Eteone longa 21 Eteone sp. 1 Eteone spilotus 11 Eulalia californiensis Eulalia sp. Eumida longicornuta Eusyllis sp. Exogone spp. 1 Glycera americana Glycera nana 3123365 4 5 14522364 Glycera robusta 1 Glycinde armigera 232 1 221
Page 35 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Goniada brunnea 11 Harmothoe imbricata Cmplx. Hesionidae Indet. Lepidasthenia berkeleyae 11 Lumbrineridae Indet. Lumbrineris californiensis 16 5 1 1 Lumbrineris cruzensis 20 20 16 18 8 17 4 Lumbrineris japonica Lumbrineris latreilli 1 15 1 Lumbrineris spp. 54 6 Malmgreniella macginitiei 11 61 Malmgreniella nigralba Nephtys cornuta 1 Nephtys ferruginea 23 131313 Nereis procera 51 5 1 1 Odontosyllis phosphorea 1 Parandalia fauveli 1 12 Pholoe glabra 11111 Pholoides asperus 18 1 11 1 1 Phyllodoce groenlandica 111 Phyllodoce hartmanae Phyllodoce sp. 1 Pilargis berkeleyae 212 3 Podarkeopsis glabrus 11 Polynoidae Indet. Scoletoma luti 694 4 3165 Sigambra bassi 11 Sige sp. Sphaerodoropsis sphaerulifer 1 Sthenelais berkeleyae Syllis elongata 1 9 2231 Syllis sp.
Page 36 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Tenonia priops 1 Typosyllis alternata 15 4 1 4 Polychaeta Sedentaria Amaena occidentalis Ampharete finmarchica 2 11 Ampharete labrops Ampharete sp. 2 Ampharetidae Indet. Aphelochaeta glandaria 22 1 221 Aphelochaeta monilaris 111 1 114 Aphelochaeta sp. 2 Aricidea lopezi 1112 Armandia brevis Artacama coniferi Barantolla americana 2 Boccardiella hamata Capitella capitata Cmplx. 2 Chaetozone setosa 21 11 Chaetozone sp. Chone sp. 1 Cirratulidae Indet. Cossura pygodactylata 1 Decamastus gracilis 104 2 36 13 6 57 57 63 19 Dipolydora cardalia 3 Dipolydora quadrilobata Dipolydora socialis 9 34 30 39 Dipolydora sp. Euchone analis 2 Euclymene sp. Euclymeninae Indet. 161 Galathowenia oculata 107 70 53 204 34 29 34 27 Heteromastus filiformis
Page 37 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Heteromastus filobranchus Heteromastus spp. Lanassa gracilis 212 Laonice cirrata 11 Leitoscoloplos pugettensis 11 Levinsenia gracilis 226 1 Lumbriclymeninae Indet. Magelona longicornis 831 41 Maldane sarsi 1 Mediomastus spp. 9 80 16 10 40 7 35 2 Megalomma splendida 11 3 1 Metasychis disparidentata 1 Mugga wahrbergi 1 Neosabellaria cementarium 1 Nichomache lumbricalis 812 1 Nichomache personata Notomastus hemipodus 11 5 1 11 20 29 3 Notomastus sp. 3 Notomastus tenuis 12 3 Notoproctus pacificus 19 Onuphis iridescens 141 5 23245343 Ophelina acuminata 1 Owenia fusiformis 1 Paraprionospio alata 3151 4 11 11 Pectinaria californiensis 13 32 1 Pectinaria granulata 21 Petaloproctus borealis 4 Pherusa sp. 13 Phylo felix 113 Pista agassizi 1 Pista brevibranchiata 1 Pista sp. 4 22
Page 38 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Pista wui 111 Polycirrus sp. 2 1 30 411 Polydora spp. 1 Polydora websteri Praxillella gracilis 1 Praxillella pacifica 12 2 Praxillella praetermissa 21 Praxillella sp. 1111 Prionospio (Minuspio) lighti 54 3 4112 Pionospio (Minuspio) multibranchiata 21 Prionospio dubia 13 Prionospio jubata 443 2 1 1 2 Pseudopolydora kempi Rhodine bitorquata 8 531 Rhynchospio glutaea Sabellidae Indet. Scalibregma californicum 13 3 33510 Scionella japonica Scoloplos acmeceps 341 615 6418 Spio cirrifera 243 2 3 1924142 Spiochaetopterus costarum 1111 Spiophanes berkeleyorum 2223 2 1 1 1 Spiophanes fimbriata 112 2 1 Spiophanes sp. Sternaspis fossor 1 Streblosoma pacifica 1 Terebellidae Indet. Terebellides californica 11 12 Terebellides sp. 2 Thelepus sp. 2 Trochochaeta multisetosa 111 Oligochaeta
Page 39 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Tubificidae SPIUNCULA Nephasoma diaphanes Thysanocardia nigra 112 111 MOLLUSCA Aplacophora Chaetoderma argenteum Chaetoderma elegans 73 Chaetoderma sp. 3 Polyplacophora Ischnochitonidae Indet. Gastropoda Alvania rosana 2 Astyris gausapata 5 Corambe sp. 1 Cylichna attonsa Euspira sp. Gastropoda Indet. (dissolved) 7 Lottidae Indet. Nassarius mendicus Odostomia sp. 1 Turbonilla sp. 4 111 Turridae Indet. Bivalvia Adontorhina cyclia 1 Axinopsida serricata 314 527 1 5 3121 4135 Cardiomya pectinata 11 Chlamys sp. Compsomyax subdiaphana 1 2 12321111 Crenella decussata 1 Cyclocardia ventricosa 1 1 Delectopecten vancouverensis 11
Page 40 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Ennucula tenuis 1 Lasaeidae Indet. Lucinoma annulatum 11 3 7 321111514 Lyonsia californica 11 Macoma carlottensis 256 3 7 55 5 Macoma elimata 2 121 3 38372124 Macoma golikovi 1217 Macoma spp. 3168 5 5769 Macoma yoldiformis 1 1 Megayoldia thracaeiformis Mya sp. 111 Mya truncata Naeromya sp. Nemocardium centifilosum 21 1 Nutricola lordi 1 Parvilucina tenuisculpta 19 103 8 75 8 94 14 46 18 49 15 56 21 143 Rochefortia tumida 12 3 Solamen columbianum 2211 Tellina sp. 1 Thyasira flexuosa 12 Yoldia seminuda Yoldia sp. Scaphopoda Antalis pretiosum Pulsellum salishorum ARTHROPODA Amphipoda Americhelidium shoemakeri Ampelisca lobata 111 Ampelisca sp. Aoroides columbiae 3 Aoroides sp.
Page 41 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Bathymedon pumilis 111 2 53 Bruzelia tuberculata Byblis millsi 111 1 Desdimelita desdichada Dyopedos arcticus Eobrolgus chumashi Guernea reduncans 2 1 Heterophoxus affinis 91522 Heterophoxus conlonae 1 Heterophoxus ellisi 112 1 11 Leucothoidae Indet. Melita oregonensis 2 Monoculodes perditus 1 Oedicerotidae Indet. 5 Opisa tridentata 3 Orchomene sp. Pachynus barnardi 1 Parametaphoxus quayleyi 244 8 322 Photis sp. Pleustidae Indet. Pleusymtes subglaber Protomedeia sp. 1 Rhachotropis barnardi 7 Rhepoxynius tridentatus Westwoodilla tone 3 1 10 1 9 1 5 1 Copepoda Harpacticoida Tanaidacea Leptochelia savignyi 3 Scolaura phillipsi 153 Ostracoda Cylindroleberidae 1
Page 42 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Euphilomedes carcharodonta 3 12 Euphilomedes producta 30 3 13 4 16 20 5 3 6 Rutiderma lomae 41 12 Scleroconcha trituberculata Cumacea Campylaspis canaliculata Campylaspis sp. 1 Diastylis santamariensis Diastylis sp. Eudorella pacifica 11 Eudorellopsis longirostris 313 Leucon sp. Leucon subnasica 1 Decapoda Crangonidae 1 Hemigrapsus sp. Oregonia gracilis 1 Pinnixa sp. 24 2 2 PHORONIDA Phoronis muelleri 7 17 1 23 15 20 24 BRYOZOA Bowerbankia gracilis colony frag. 1 Tubuliporidae colony frag. 1 ECHINODERMATA Ophiuroidea Amphiodia urtica 12 1321 Ophiura sarsi 32 Ophiura sp. Holothuroidea Pentamera populifera 1 6 Pentamera pseudocalcigera 11 Pentamera rigida 11
Page 43 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB6 CRB6 CRB6 CRB6A CRB6A CRB6A CRB6A CRB6A CRB6A CRB6B CRB6B CRB6B CRB6B CRB6B CRB6B CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/12/09 3/10/09 Replicate 233112 2331122331 Field Sieve 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad Juv Ad HEMICHORDATA Saccoglossus sp. 65
Notes: Calanoid 1 Gastropod egg cases Nematoda 34 Leptomedusae (clear jellyfish)
Page 44 Crofton 2009 Benthic Invertebrate Data Prepared by COLUMBIA SCIENCE Prepared for Hatfield Consultants, Ltd. DATA REPORT CROFTON 2009
Submitting Firm Submitter's Name Address Phone/Fax Sample State Description Sample Preservation Dates Analyzed Taxonomist
Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv
CNIDARIA Hydrozoa Campanulariidae colony frag. Corymorphidae Indet. Monobrachium parasitum colony frag. Perigonimus repens colony fragment 11 Anthozoa Halcampa decemtentaculata Halipteris californica Metridium farcimen Pachycerianthus fimbriatus Pennatulacea Indet. Virgularia sp. PLATYHELMINTHES Stylochidae Indet.
Page 45 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv NEMERTEA Anopla Indet. Carinoma mutabilis Cerebratulus californiensis 111 Hoplonemertea Indet. Lineidae Lineus sp. Palaeonemertea Paranemertes californica Tubulanidae Tubulanus nothus 1 Tubulanus polymorphus 71 ANNELIDA Polychaeta Errantia Arctonoe sp. Dorvillea longicornis Dorvillea sp. Driloneris longa Errano bicirrata Eteone californica Eteone longa 1 Eteone sp. Eteone spilotus Eulalia californiensis Eulalia sp. Eumida longicornuta Eusyllis sp. Exogone spp. Glycera americana Glycera nana 38451 Glycera robusta Glycinde armigera 211
Page 46 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Goniada brunnea Harmothoe imbricata Cmplx. Hesionidae Indet. Lepidasthenia berkeleyae Lumbrineridae Indet. 1 Lumbrineris californiensis Lumbrineris cruzensis 15 3 Lumbrineris japonica Lumbrineris latreilli 3 Lumbrineris spp. Malmgreniella macginitiei Malmgreniella nigralba Nephtys cornuta Nephtys ferruginea 2 Nereis procera 11 4 Odontosyllis phosphorea Parandalia fauveli Pholoe glabra 1 Pholoides asperus Phyllodoce groenlandica Phyllodoce hartmanae Phyllodoce sp. Pilargis berkeleyae 11 Podarkeopsis glabrus Polynoidae Indet. Scoletoma luti 54 Sigambra bassi Sige sp. Sphaerodoropsis sphaerulifer Sthenelais berkeleyae Syllis elongata Syllis sp.
Page 47 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Tenonia priops Typosyllis alternata 2 Polychaeta Sedentaria Amaena occidentalis Ampharete finmarchica Ampharete labrops Ampharete sp. Ampharetidae Indet. Aphelochaeta glandaria Aphelochaeta monilaris Aphelochaeta sp. Aricidea lopezi Armandia brevis Artacama coniferi Barantolla americana Boccardiella hamata Capitella capitata Cmplx. Chaetozone setosa 2 Chaetozone sp. Chone sp. Cirratulidae Indet. 1 Cossura pygodactylata Decamastus gracilis 11 42 16 20 4 Dipolydora cardalia Dipolydora quadrilobata Dipolydora socialis 9 Dipolydora sp. Euchone analis Euclymene sp. Euclymeninae Indet. 512 Galathowenia oculata 31 30 Heteromastus filiformis
Page 48 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Heteromastus filobranchus Heteromastus spp. Lanassa gracilis Laonice cirrata Leitoscoloplos pugettensis 212 Levinsenia gracilis Lumbriclymeninae Indet. Magelona longicornis 1 Maldane sarsi Mediomastus spp. 29 5 Megalomma splendida Metasychis disparidentata Mugga wahrbergi Neosabellaria cementarium Nichomache lumbricalis Nichomache personata Notomastus hemipodus 36 Notomastus sp. Notomastus tenuis Notoproctus pacificus Onuphis iridescens 21 Ophelina acuminata 1 Owenia fusiformis Paraprionospio alata 11 1 Pectinaria californiensis 2 Pectinaria granulata Petaloproctus borealis Pherusa sp. Phylo felix 31 Pista agassizi 1 Pista brevibranchiata Pista sp.
Page 49 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Pista wui Polycirrus sp. 122 1 Polydora spp. Polydora websteri Praxillella gracilis Praxillella pacifica Praxillella praetermissa Praxillella sp. 1 Prionospio (Minuspio) lighti 2 Pionospio (Minuspio) multibranchiata Prionospio dubia 11 Prionospio jubata 1411 Pseudopolydora kempi Rhodine bitorquata Rhynchospio glutaea Sabellidae Indet. Scalibregma californicum 67342 Scionella japonica Scoloplos acmeceps 510798 Spio cirrifera 22 Spiochaetopterus costarum 1 Spiophanes berkeleyorum 2 123 Spiophanes fimbriata Spiophanes sp. Sternaspis fossor Streblosoma pacifica Terebellidae Indet. Terebellides californica 1111 Terebellides sp. Thelepus sp. Trochochaeta multisetosa Oligochaeta
Page 50 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Tubificidae SPIUNCULA Nephasoma diaphanes Thysanocardia nigra MOLLUSCA Aplacophora Chaetoderma argenteum Chaetoderma elegans Chaetoderma sp. Polyplacophora Ischnochitonidae Indet. Gastropoda Alvania rosana Astyris gausapata Corambe sp. Cylichna attonsa 11 Euspira sp. Gastropoda Indet. (dissolved) Lottidae Indet. Nassarius mendicus Odostomia sp. Turbonilla sp. 31 Turridae Indet. Bivalvia Adontorhina cyclia Axinopsida serricata 4 38 6 26 6 Cardiomya pectinata 11 Chlamys sp. Compsomyax subdiaphana 12213 Crenella decussata Cyclocardia ventricosa 1 Delectopecten vancouverensis
Page 51 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Ennucula tenuis Lasaeidae Indet. Lucinoma annulatum 1 13 2 16 4 Lyonsia californica 1 Macoma carlottensis 47 Macoma elimata 34353 Macoma golikovi 11 Macoma spp. 557 Macoma yoldiformis Megayoldia thracaeiformis Mya sp. Mya truncata 2 Naeromya sp. Nemocardium centifilosum 12 Nutricola lordi Parvilucina tenuisculpta 30 234 55 195 39 Rochefortia tumida 4 Solamen columbianum 11 2 Tellina sp. Thyasira flexuosa 43 Yoldia seminuda Yoldia sp. Scaphopoda Antalis pretiosum Pulsellum salishorum 31 ARTHROPODA Amphipoda Americhelidium shoemakeri Ampelisca lobata Ampelisca sp. 2 Aoroides columbiae Aoroides sp.
Page 52 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Bathymedon pumilis Bruzelia tuberculata Byblis millsi 1 Desdimelita desdichada Dyopedos arcticus Eobrolgus chumashi Guernea reduncans Heterophoxus affinis Heterophoxus conlonae Heterophoxus ellisi Leucothoidae Indet. Melita oregonensis Monoculodes perditus Oedicerotidae Indet. Opisa tridentata Orchomene sp. 1 Pachynus barnardi Parametaphoxus quayleyi 1 Photis sp. 1 Pleustidae Indet. Pleusymtes subglaber Protomedeia sp. 1 Rhachotropis barnardi Rhepoxynius tridentatus 2 Westwoodilla tone Copepoda Harpacticoida Tanaidacea Leptochelia savignyi Scolaura phillipsi Ostracoda Cylindroleberidae
Page 53 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv Euphilomedes carcharodonta Euphilomedes producta 12 6 Rutiderma lomae Scleroconcha trituberculata Cumacea Campylaspis canaliculata Campylaspis sp. Diastylis santamariensis Diastylis sp. Eudorella pacifica Eudorellopsis longirostris Leucon sp. Leucon subnasica Decapoda Crangonidae Hemigrapsus sp. Oregonia gracilis Pinnixa sp. 1 PHORONIDA Phoronis muelleri 326 BRYOZOA Bowerbankia gracilis colony frag. Tubuliporidae colony frag. ECHINODERMATA Ophiuroidea Amphiodia urtica 213 Ophiura sarsi 1 Ophiura sp. Holothuroidea Pentamera populifera 1 Pentamera pseudocalcigera Pentamera rigida
Page 54 Crofton 2009 Benthic Invertebrate Data Hatfield Station ID CRB7 CRB7 CRB7 CRB7 CRB7 Sampling Date (m/d/y) 3/10/09 3/10/09 3/10/09 3/10/09 3/10/09 Replicate 12233 Field Sieve 1mm 1mm 1mm 1mm 1mm TAXA Juv Ad Juv Ad Juv HEMICHORDATA Saccoglossus sp.
Notes: Calanoid Gastropod egg cases Nematoda Leptomedusae (clear jellyfish)
Page 55 Crofton 2009 Benthic Invertebrate Data Columbia Science Sorting QA
Project: Hatfield Consultants - Crofton 2009 Date: March 9-12, 2009 Sampling Dates
# from 20% Resort 50% Resort 100% Resort Initial sort Resort Final Sample ID Date d/m/y First sort # found X 5 OR # found X 2 OR # found X 1 Efficiency Required? Efficiency CRB 1A-1 03/12/09 964 24 97% N 97% CRB 1A-2 03/12/09 814 0 100% N 100% CRB 1A-3 03/12/09 1125 0 100% N 100% CRB 2-1 03/09/09 660 15 98% N 98% CRB 2-2 03/09/09 1199 30 98% N 98% CRB 2-3 03/09/09 1088 50 96% N 96% CRB 2A-1 03/12/09 787 3 99% N 99% CRB 2A-2 03/12/09 647 5 99% N 99% CRB 2A-3 03/12/09 1308 0 100% N 100% CRB 3-1` 03/11/09 943 0 100% N 100% CRB 3-2 03/11/09 796 0 100% N 100% CRB 3-3 03/11/09 768 30 96% N 96% CRB 4A-1* 03/11/09 987 0 100% N 100% CRB 4A-2 03/11/09 1235 0 100% N 100% CRB 4A-3** 03/11/09 1274 0 100% N 100% CRB 5A-1 03/10/09 475 20 96% N 96% CRB 5A-2 03/10/09 565 14 98% N 98% CRB 5A-3 03/10/09 538 0 100% N 100% CRB 5B-1 03/09/09 701 20 97% N 97% CRB 5B-2 03/09/09 529 0 100% N 100% CRB 5B-3 03/09/09 587 0 100% N 100% CRB 5C-1 03/10/09 302 0 100% N 100% CRB 5C-2 03/10/09 230 0 100% N 100% CRB 5C-3 03/10/09 398 0 100% N 100%
CRB 6-1 03/10/09 633 25 96% 96% CRB 6-2 03/10/09 377 0 100% N 100% CRB 6-3 03/10/09 598 17 97% N 97% CRB 6A-1 03/12/09 530 0 100% N 100% CRB 6A-2 03/12/09 377 10 97% N 97% CRB 6A-3 03/12/09 662 8 99% N 99% CRB 6B-1 03/10/09 457 2 99% N 99% CRB 6B-2 03/10/09 397 0 100% N 100% CRB 6B-3 03/10/09 495 0 100% N 100% CRB 7-1 03/10/09 428 0 100% N 100% CRB 7-2 03/10/09 663 0 100% N 100% CRB 7-3 03/10/09 476 0 100% N 100%
Note: At least 20% of the total number of samples (36 x .2 = 7) were re-sorted at 100% and all others at 20% or 50% * Sample 4A-1 split in half for sorting and analysis -QA performed on sorted half ** Sample 4A-3 split in 1/4 for sorting and analysis - QA performed on sorted 1/4 Subsampling Tests - Crofton 2009 Tests for Precision Sample ID Subsample Number Expected Actual Difference % Difference Subsample CRB 1A-3 1 291 1164 1133 31 2.7 1234 CRB 1A-3 2 277 1108 1133 -25 -2.2 Count 291 277 286 279 CRB 1A-3 3 286 1144 1133 36 3.1 Precision 4.81 3.15 2.45 CRB 1A-3 4 279 1116 1133 -53 -4.6 1.72 0.72 Mean subsampling error 3.1% 4.12 Note: Each of 3 jars split separately for total of 12 splits. 1 split from each jar combined for total Min % error 2.2 Range of Precision 0.72% to 4.81% Max % error 4.6 CRB 2A-1 1 205 820 810 10 1.2 Count 205 212 202 191 CRB 2A-1 2 212 848 810 38 4.7 Precision 3.3 4.72 5.45 CRB 2A-1 3 202 808 810 -2 0 1.46 9.91 CRB 2A-1 4 191 764 810 -46 -5.7 6.83 Mean subsampling error 2.9% Note: 1 jar Min. % error 0 Range of Precision 1.46% to 9.91% Max % error 5.7 CRB 2A-2 1 149 596 564 32 5.7 In this sample I did not remove the Count 149 132 147 136 CRB 2A-2 2 132 528 564 -68 -12.1 Parvilucina prior to splitting and they Precision 11.41 10.2 7.48 CRB 2A-2 3 147 588 564 92 16.3 were not distributed evenly. 1.34 2.94 CRB 2A-2 4 136 544 564 -20 -3.5 I did not go back because this sample 8.73 was put into one jar for analysis anyway. Mean sampling error 9.4% Range of Precision 1.34% to 11.41% Note: 1 jar Min. % error 3.5 Max % error 16.3 CRB 2A-3 1 336 1344 1422 78 -5.5 Count 336 386 340 360 CRB 2A-3 2 386 1544 1422 122 8.6 Precision 12.95 11.92 5.56 CRB 2A-3 3 340 1360 1422 62 -4.4 1.18 6.74 CRB 2A-3 4 360 1440 1422 80 5.6 6.67 Mean sampling error 6.0% Note 2 jars split separately and 1 split from each combined for total. Min % error 4.4 Range of Precision 1.18% to 12.95% Max % error 8.6 Crust VOUCHER VERIFICATIONS
ORIGINAL TAXONOMY: COLUMBIA SCIENCE-SANDY J LIPOVSKY
PROJECT: CROFTON 2009
TAXONOMIC CATEGORY: CRUSTACEA VERIFICATION BY: BIOLOGICA ENVIRONMENTAL SERVICES, LTD BIOLOGICA TAXONO Phil Hoover VERIFICATION DATE: November 2009
HATFIELD STATION TAXA # QA Identification COMMENTS Note: Only A and J for this project. Purple caps: 6-1 Scolaura phillipsi 2A ID Confirmed 4A-2 Rhepoxynius tridaentatus 1A ID Confirmed 6-1 Byblis millsi 1A ID Confirmed 2-1 Bruzelia tuberculata 3A ID Confirmed 6-1 Euphilomedes producta 3A ID Confirmed 6-1 Eudorellopsis longirostris 3A ID Confirmed 6-1 Heterophoxus ellisi 1A ID Confirmed 6-1 Heterophoxus conlonae 1A ID Confirmed Spelling mistake H. conlanae 6-1 Guernea reduncans 2A ID Confirmed 6-1 Parametaphoxus quayleyi 4A ID Confirmed 1A-2 Aoroides columbiae 4A Aoroides intermedius Setae on gnathopod 1 segment 5 appears wrong for columbiae 2-1 Heterophoxus affinis 2A ID Confirmed 5B-1 Campylaspis canaliculata 1A ID Confirmed
Page 1 Misc
ORIGINAL TAXONOMY: COLUMBIA SCIENCE-SANDY J LIPOVSKY
PROJECT: CROFTON 2009
TAXONOMIC CATEGORY: MISCELLANEOUS VERIFICATION BY: BIOLOGICA ENVIRONMENTAL SERVICES, LTD BIOLOGICA TAXONOMIST: Valerie Macdonald VERIFICATION DATE: November 2009
HATFIELD BIOLOGICA STATION ID TAXA # QA Identification COMMENTS Note: Only A and J designations for this project Large vials: 5C-2 Pachycerianthus fimbriatus 1A ID Confirmed Adult Purple caps: 5A-2 Corymorphidae 1A Pinauay crocea (Tubularidae) Shaped gonophores are on a stem positioned above whorl of tentacles not below whorl. Stem does not have striations as in Corymorpha palma. 6-1 Lineidae (?) 2 Juv Cerebratulus californiensis Presence of cephalic slits, position of mouth and caudal cirrus. Suggest these are large juveniles as no reproductive evidence is apparent. 5A-3 Tubulanus nr. nothus 3 Juv Tubulanus polymorphus Presence of white band anterior to dark pigment band is characteristic of T. polymorphus.
Page 2 Moll
VOUCHER VERIFICATIONS
ORIGINAL TAXONOMY: COLUMBIA SCIENCE-SANDY J LIPOVSKY
PROJECT: CROFTON 2009
TAXONOMIC CATEGORY: MOLLUSCS VERIFICATION BY: BIOLOGICA ENVIRONMENTAL SERVICES, LTD BIOLOGICA TAXONOMIST: Katie Baumann VERIFICATION DATE: November 2009
HATFIELD BIOLOGICA STATION ID TAXA # QA Identification COMMENTS Purple caps: 7-2 Solamen columbianum 1J ID Confirmed
Page 3 Poly
VOUCHER VERIFICATIONS
ORIGINAL TAXONOMY: COLUMBIA SCIENCE-SANDY J LIPOVSKY
PROJECT: CROFTON 2009
TAXONOMIC CATEGORY: POLYCHAETES VERIFICATION BY: BIOLOGICA ENVIRONMENTAL SERVICES, LTD BIOLOGICA TAXONOMIST: Hiroki Tomoe VERIFICATION DATE: November 2009
HATFIELD BIOLOGICA STATION ID TAXA # QA Identification COMMENTS Note: For this project only A and J designations Large vial/jar 5C-1 Glycera robusta 1A ID Confirmed 2-3 Glycera americana 1A ID Confirmed 6A-2 Praxillella gracilis 1A ID Confirmed 5B-1 Artacama coniferi 1A ID Confirmed 5B-3 Metasychis disparidentata 1A Chirimia biceps Lobe of pygidial plate was not long but short. Also, dorsal lobe of cephalic plate short with many teeth. Purple caps: 6-1 Rhodine bitorquata 1A ID Confirmed 6-1 Prionospio (Minuspio) multibranchiata 2A ID Confirmed 6A-3 Euchone analis 2A ID Confirmed 4A-2 Notomastus tenuis 4A ID Confirmed 6-1 Scalibregma californicum 6A ID Confirmed 2-1 Cossura pygodactylata 2A ID Confirmed 4A-2 Eteone spilotus 4A ID Confirmed 6-1 Ophelina acuminata 1A ID Confirmed 6-1 Phylo felix 1A ID Confirmed 6B-3 Aphelochaeta monilaris 2A Aphelochaeta nr. tigrina Shape of neurosetae as in Santa Barbara Vol 6 5A-2 Chaetozone setosa 1A ID Confirmed 6-1 Nichomache personata 1A ID Confirmed 6-1 Dipolydora socialis 4A ID Confirmed 2A-1 Dipolydora quadrilobata 2A Dipolydora caulleryi Major spines, large flattened, beaklike curved end bearing large and distinctive crest of bristles.
Page 4 Poly
5B-3 Boccardiella hamata 1A ID Confirmed 5A-1 Terebellides californica 1A ID Confirmed 6A-3 Neosabellaria cementarium 1A ID Confirmed
Page 5
Appendix A8
Benthic Survey: Power Analysis for Regressions
Figure A8.1 Post-hoc power analysis for regressions to detect critical effect r ≥ |0.707|.
Appendix A9
Benthic Survey: Redox Potential and Sulphides Preparation, Calibration, and Analysis
MINISTRY OF WATER, LAND AND AIR PROTECTION
Protocols for Marine Environmental Monitoring
05 September 2002
Assistant Deputy Minister
Environmental Protection Division Table of Contents
Introduction ...... 3
Acronyms, Abbreviations, & Definitions...... 4
1. Currents Metering...... 6 Equipment ...... 6 Procedures...... 6 Reporting...... 6 2. Video Surveys...... 9 Equipment ...... 9 Procedures...... 9 3. Sediment Sampling...... 11 Equipment ...... 11 Procedures...... 11 Reporting...... 14 4. Checking the Quality of Sediment Samples...... 15 Physical and Chemical QA/ QC...... 15 Biological QA/ QC...... 15 Reporting...... 15 5. Calibrating the Sulphide Electrode ...... 16 Materials ...... 16 Preparing Solutions...... 16 Procedures...... 18 6. Standardizing the Redox Electrode...... 20 Materials ...... 20 Procedures...... 20 7. Performing Statistical Analyses...... 21 Preparations for Statistical Analyses ...... 21 Statistical Methods to Determine If Requirements Have Been Met ...... 22 Statistical Power Analyses ...... 25 Appendix A: Design of Video Survey...... 26 Baseline Monitoring...... 26 Appendix B: Design of Sediment Sampling...... 27 Baseline Monitoring...... 27 Operational Monitoring...... 27 Appendix C: Statistical Procedures ...... 29
Protocols for Marine Environmental Monitoring 2 Introduction
To support the Finfish Aquaculture Waste Control Regulation, the Ministry of Water, Land and Air Protection (WLAP) has developed Protocols for Marine Environment Monitoring. These protocols will ensure that high quality data are collected, thereby leading to sound decisions as to whether environmental standards are being met. WLAP developed the protocols with assistance from various government agencies, consultants, literature reviews, and equipment manufacturers.
The protocols were developed with regard to the aquaculture industry. However, these protocols may also be relevant to monitoring or assessing impacts of other anthropogenic activities.
The protocols comprise 7 sections:
Section 1 lists acceptable types of current meters for generating data on currents at BC aquaculture operations. It also specifies the supporting information that monitoring agencies must submit.
Section 2 specifies the video equipment for completing video surveys (typically of hard-bottom sites) and outlines procedures for deploying the camera and generating acceptable quality video.
Section 3 describes materials and methods for soft-bottom sampling, including procedures for obtaining specific types of data.
Section 4 outlines the quality assurance/ quality control requirements for physical and chemical parameters and biological samples.
Sections 5 and 6 describe procedures for standardizing and calibrating field meters for sulphide and Eh measurements. The procedures are specific to ThermoOrion meters and probes, the most widely used brand. Other companies’ meters and probes are acceptable, provided the standardization and calibration procedures provided by the manufacturer are followed.
Section 7 describes statistical tools for analysing sampling data from soft-bottom sites and the video from hard-bottom sites.
Appendix A summarizes the video survey requirements for baseline inventory monitoring.
Appendix B summarizes the sediment sampling requirements for both baseline inventory and operational monitoring.
Appendix C describes statistical procedures to be used for existing facilities and new facilities respectively.
Protocols for Marine Environmental Monitoring 3 Acronyms, Abbreviations, & Definitions
ANOSIM: analysis of similarities
ANOVA: analysis of variance
BACI: before-after-control-impact (study design)
Baseline monitoring: sampling conducted before operation of a finfish aquaculture facility
BCGS: British Columbia Geographic System
Beggiatoa: a genus of bacteria that forms white mats on the sediment surface in areas of intense organic enrichment
Capitella: a genus of polychaetes that thrives in areas of intense organic enrichment
CEAA: Canadian Environmental Assessment Act
Cu: copper concentration (expressed in µg/ g dry sediment)
DGPS: Differential Global Positioning System
DI: de-ionized
EDTA: ethylenediaminetetraacetic acid
Eh: redox potential (expressed in millivolts, mV)
Epifauna: animals that live on top of the substratum
EScrit: critical effect size
HA: alternate hypothesis
HO: null hypothesis
Infauna: animals that live within the substratum
LWBC: Land & Water British Columbia Inc.
M: median
Macrofauna: animals with body sizes on the scale of millimetres
MAFF: BC Ministry of Agriculture, Fisheries and Food
MCI: multiple control/ impact (study design)
MDS: multi-dimensional scaling
Protocols for Marine Environmental Monitoring 4 Megafauna: animals with body sizes on the scale of centimetres
MLR: multiple linear regression
N: sample size
NAD: North American datum
NLR: non-linear regression
Operational monitoring: sampling conducted during operation of a finfish aquaculture facility and as outlined in Schedule B of the Finfish Aquaculture Waste Control Regulation
QA/ QC: quality assurance/ quality control
ROV: remotely operated vehicle
S=: free sulfide concentration (expressed in micromolar, µM)
SAOB: sulphide anti-oxidant buffer
SD: standard deviation
SGS: sediment grain size
SLR: simple linear regression
TOC: total organic carbon (expressed in µg/ g dry sediment)
TVS: total volatile solids (expressed as a percentage)
WLAP: BC Ministry of Water, Land and Air Protection
x : sample mean
Zn: zinc concentration (expressed in µg/ g dry sediment)
1 – ß: power (of statistical test) a: Type I error rate (significance level)
ß: Type II error rate
ß1: rate of increase (non-linear) or slope (linear) of population regression line
µ: population mean
Protocols for Marine Environmental Monitoring 5
1. Currents Metering
Equipment Electronic current meters capable of determining both speed and direction are available from several manufacturers (Aanderaa, Sontek, Nortek, RD Instruments, Applied Microsystems, InterOcean Systems, etc). We recommend a meter with an internal data-logger, which can be pre-set for the correct interval and record the results automatically. Both vector-averaging and instantaneous type meters are acceptable. Only experts should attempt to program and deploy these devices, or extract and process the collected data.
Procedures 1. Measure currents at 2 depths: approximately 15 m below the surface and approximately 5 m above the bottom.
2. Report current direction in degrees True (include magnetic north reading and correction factor) and speed in cm/ s.
3. Record current speed and direction at least once every 30 min over a period of at least 30 days.
4. At sites with infrastructure in place, locate the meter away from attenuation effects of any infrastructure and in line with the prevailing current direction. Moor the current meter approximately 30 m from the offshore side of the containment structure unless circumstances do not allow it.
5. At sites where infrastructure has not yet been installed, metering locations should represent currents within the tenure, especially near containment structures.
Reporting This information must be included with both the raw data and data summaries:
1. Current meter mo orings and deployment locations Supply a diagram showing how the current meters were deployed within the tenure area. Include in the diagram: · the type and position (surface or sub-surface) of the flotation devices used to support the current meters during deployment · the distances between the current meter and the flotation device · the type and w eights of anchors used. Also, show and describe any other components or instruments attached to the mooring apparatus (e.g. mechanical or acoustic releases). Supply DGPS co-ordinates for the deployment locations and a written description of the locations (e.g. 30 m at 270º from the southwest corner of the containment structures), indicating the locations on a map. A 1:20,000 scale BCGS map is recommended, but equivalents are acceptable. Indicate whether the DGPS co-ordinates and maps are based on the NAD 27 or NAD 83 co-ordinate system.
Protocols for Marine Environmental Monitoring 6 2. Start date and time Record the date and time that the current monitoring commenced (i.e. the individual date and time that each meter began to collect and record good-quality data of local currents). Indicate whether time is recorded as Pacific Standard Time (UTC-8) or Pacific Daylight Saving Time (UTC-7).
3. End date and time Record the date and time that the current monitoring was terminated for each meter (i.e. the date and time the current meter collected its last good record of the currents before it was recovered). Clearly indicate whether time is recorded as above.
4. Instrument Provide the make and model of the current me ters used, including a copy of the manufacturer’s specifications, and date of last calibration and servicing.
5. Number of data points Report the actual number of instantaneous or average measurements recorded by the meter. If measurements are taken every 30 min, there will be approximately 1400 measurements in the monitoring period and therefore 1400 data points. This number assists in calculating averages.
6. Sample interval Report the sample interval (min) between consecutive measurements made by the meter. A sample interval must be 30 min or less.
7. Data processing and reporting Describe the data-processing methods and software used to correct and process the current meter data. Indicate whether the current direction is in degrees True (recommended) or degrees magnetic. Indicate whether the current meter records average or instantaneous measurements, and describe the instrument’s set-up or configuration. If the meter records average measurements, indicate the averaging interval. Details should be provided in distinct sections of the report under the appropriate section headings or titles.
8. Depth of meter Report the depth of the meter below the water surface or the distance of the meter from the bottom. The meter should be 15 m below the surface for surface-currents measurements and approximately 5 m above bottom for bottom-currents measurements.
9. Water depth Report the water depth at the location of deployment.
10. Average current speed Calculate the average current speed for the entire data-collection period (30 d). This should be calculated from the entire dataset, not from the summary data.
Protocols for Marine Environmental Monitoring 7 11. Contact names Provide the name and contact information of the staff person or consulting company responsible for collecting and reporting the current measurements.
Protocols for Marine Environmental Monitoring 8
2. Video Surveys
Equipment Acceptable vehicles for carrying video equipment include:
· An ROV
· A cable camera apparatus
· Scuba divers
Video equipment must meet these criteria:
· be capable of producing broadcast-quality images
· have supplemental light to increase clarity and maintain good colour balance
· have a reference object or superimposed image to show scale on the viewing screen in metres
· have an in-built DGPS unit or similar tracking device to define the transect or station being videotaped
· original video must be transferable to digital-format storage media.
Acceptable quadrat types include:
· A wire frame (1 × 1 m, with nine 33 × 33 cm sections) placed on the seabed
· A wire frame mounted on the cable camera or ROV
· A laser-delineated frame
Acceptable transect lines include:
· Brightly-coloured polypropylene ropes, weighted, and with flagging tape placed at regular intervals
· Brightly-coloured measuring tapes, weighted
Procedures See Appendix A for summary of design information.
A. Baseline Monitoring
1. Survey several transects across the entire tenure, capable of mapping biophysical characteristics to a 50 m resolution. 2. Within the tenure, s urvey a minimum of one transect perpendicular to shore, starting at the shore and terminating at the opposite perimeter of the tenure to describe depth
Protocols for Marine Environmental Monitoring 9 variation. Surveys should encompass all area(s) of probable footprint(s) expected for future containment structures. 3. Survey a minimum of 2 transects at each of 2 reference stations, each 100 m long with one oriented perpendicular to shore. 4. Use a sufficient number of macrofauna quadrats to represent each substratum type. 5. Quadrats must measure 1 × 1 m, with nine 33 × 33 cm sections. Note that quadrat size must be the same at all stations. 6. Place at least 5 quadrats at each station. B. Selection of Reference Stations 1. Locate stations within a range of 0.5 – 2.0 km from facility. 2. Locate stations at least 0.5 km apart. 3. Ensure that the mean depth is within 20% of the mean depth of the facility tenure. 4. Ensure that characteristics such as topography, angle of repose, current and tidal regimes, amount of freshwater run-off, etc. are similar to those at facility stations; 5. If facility stations are potentially influenced by other human activities (e.g. log dumping), seek reference stations that may be similarly influenced. C. Deployment
1. Place transect line on bottom for camera or ROV to follow on a selected bearing. 2. Attach a small boat anchor and vertical line to one end of the transect line. The camera or ROV will use the vertical line as a guide to the transect line. 3. A DGPS reading must be taken at the beginning and end of each transect and at each quadrat. Readings associated with transects are to be taken when the transect line has been pulled taut. 4. If possible, deploy the camera during slack tide to minimize drifting. 5. Deploy the camera during daylight, when there is plenty of well-diffused light. Avoid taking videos at night or in extreme overcast conditions. 6. Keep the camera or ROV close enough to the bottom to provide optimum resolution of the bottom and never more than 1.5 m above the substratum. 7. Manoeuvre the camera or ROV at a maximum speed of 0.25 m/ s. 8. Position the transect line at the edge of the camera’s field of view so that it focuses on substratum and not on the line. 9. In areas of extreme slope and or boulder complexes, move the camera from deeper to shallower water to ensure that the field of view includes the substratum. D. Reporting
Submit data after filling out templates provided by WLAP. Provide either audio (i.e. voice dubbing) or text narration of the video. For each quadrat, describe the angle of repose as either horizontal, vertical, or oblique.
Protocols for Marine Environmental Monitoring 10
3. Sediment Sampling
Equipment 1. Acceptable sampling devices for chemical/ physical sampling include Petite Ponar, Ponar, Smith-MacIntyre, VanVeen or other appropriate equipment.
2. For biological sampling, use a Smith-MacIntyre, VanVeen, or other appropriate large-volume sediment sampling device with a 0.1 m2 footprint.
3. Various probes and chemicals, described fully in Sections 4 & 5, are to be used.
Procedures For a summary of sampling design, see Appendix B.
A. Baseline Monitoring
· Within each of the probable footprints, at least 3 grab samples must be taken for each sediment type. If only one sediment type predominates, at least 5 grab samples must be taken.
· 2 reference stations must be selected as described above for video surveys; at least 3 grabs must be taken at each reference station.
B. Operational Monitoring · Ensure the transect is parallel to the predominant current direction. · Use at least one transect for each dominant current direction or alternate design, provided extent and magnitude of effects is represented. · Sample at least 3 stations on each transect: at perimeter of the containment structure, at 30 m from zero metre station, and at perimeter of tenure. C. Selection of Reference Stations · Sample at least 2 reference stations for each facility. · Ensure the stations are within 0.5 – 2.0 km of the facility tenure, if possible. · Ensure reference stations are at least 0.5 km apart, if possible. · Ensure the mean depth is within 20% of the mean depth of facility stations. · Ensure the SGS fractions are within 15% of the facility stations’ SGS fractions. · Ensure that characteristics such as topography, current and tidal regimes, amount of freshwater run-off, etc., are similar to that of the facility stations. · If the facility stations appear to have been influenced by anthropogenic activity ensure that the reference stations have similar characteristics to that of the facility stations (e.g. log dumps).
D. Sampling Preparation 1. Prepare a sulphide stock solution and EDTA/ NaOH solution in advance. Note that:
Protocols for Marine Environmental Monitoring 11 § 10,000 µM sulphide stock solution is stable for up to 5 d, if it is kept cool with limited head space. § EDTA/ NAOH solution is stable for up to 7 d, if kept cool. 2. Check tidal conditions. If possible, do not sample during maximum flood and ebb tides or strong wind conditions.
4. Obtain latitude/ longitude using DGPS, with a minimum accuracy of ± 5 m at each station.
5. When sampling on a transect, use polypropylene rope pre-marked in metre increments to ensure accurate measurements.
6. When sampling on a transect, note bearing. Report the true-north bearing as well as the magnetic north reading and correction factor.
7. Report water depths in metres.
8. Check the Eh electrode against standard (re-check every 4 hr and when recalibrating the sulphide meter).
9. Calibrate the sulphide electrode, and recalibrate it at least every 3 - 4 hr.
10. Drift: before recalibration, and hourly during sampling or at a minimum at the completion of each sampling station, check and record the drift by measuring the sulphide concentration against each of the standard concentrations originally used to calibrate the electrode. Always use fresh standards (1000, 100, 10 µM) by serial dilution from the stock solution when recalibrating or checking drift. Do not attempt to correct the data for any observed drift. A drift of up to 20% is acceptable.
E. Collect and describe samples 1. Deploy and retrieve sampling device at a maximum rate of 0.3 m/ sec. Rinse all equipment with ambient seawater between grab deployments. Take care that 2nd and 3rd grab samples are not taken from the crater formed by the first grab sample. Typically, this is only of concern at the when the sampling vessel is moored at the edge of the containment structure.
2. Check for these indicators of an acceptable sample:
§ overlying water present – indicating minimal leakage § overlying water not excessively turbid – indicating minimal sample disturbance § sediment surface relatively flat – indicating minimal sample disturbance or washing § desired penetration depth achieved – at least 4 to 5 cm for characterizing surficial sediments § overfilled sampling device – if occurring routinely some or all of the detachable weight might have to be removed 3. Do not make more than 4 deployments of the grab to obtain a suitable sample. If unsuccessful, provide a video of the station as an alternative (see Section 2)
4. Siphon the overlying water from the sample. Retain it for sieving if biological samples required.
5. Examine the sediment sample and record the following:
Protocols for Marine Environmental Monitoring 12 § Sediment texture, colour, odour, presence/absence of gas bubbles, Beggiatoa, fish feed, fish feces, flocculent organic material, macrophytes, terrigenous material, and farm litter 6. Take a colour photo of the sample or score sediment colour by comparing with colour charts
7. Record the penetration depth of the sampler in centimetres.
= F. Measure S and Eh levels 1. Extract 2 sub-samples by removing the top 2 centimetres of sediment from the centre of each side of the sampling device. Limit the volume of each sub-sample to what is needed for the required tests as summarised in Appendix B (i.e. 50 mL required for Eh potential and sulphide concentration). Place the 2 sub-samples in a suitable container and homogenize by gently stirring with a flat tipped steel spatula. Sulphide and Eh analyses must done within 60 min of sampling to avoid sample degradation. Wear gloves if you will be touching the sediment.
2. Measure sulphide:
a. Rinse the electrode with distilled water and blot it dry. Then insert it into sample. b. When the initial sample is obtained and accepted, add 8.75 mg L ascorbic acid to 250 mL of previously prepared EDTA/ NaOH buffer and thoroughly mix to create SAOB buffer. (Various amounts of SAOB can be made, provided the 8.75 g L ascorbic acid: 250 mL EDTA/ NaOH ratio is maintained). c. Combine equal volumes of sediment and SAOB in a suitable container (5 mL of each is typically sufficient for this analysis). The sediment from the sample can extracted using a cut-off syringe or spatula. Do not include material more than 0.5 cm in diameter. d. Homogenize the mixture with a spatula. e. Insert the sulphide electrode into the solution and gently swirl it until meter reads READY (typically 2 – 5 min). f. Gently wipe the probe before the next sample. If an oily residue is observed on the probe, wash it with detergent before taking another sample. 3. Obtain an Eh measurement. Insert the probe into the homogenized sample described above , and wait until either the meter reads READY or the drift is 3 mV or less over a 2 sec period. Gently wipe excess sediment from the probe between sample measurements.
4. Record the temperature in the sediment sample.
5. Correct the Eh measurements by using sediment temperature and the correction factor for the filling solution in the probe supplied by the manufacturer.
6. Perform any additional measurement or analysis using the sediment sub-samples collected in Section C, step 1. Do not collect additional sub-samples for these analyses. Remove all unrepresentative material (e.g. shells, large worms, wood waste, rock) before filling the sampling receptacle See Appendix A for sample frequency and location.
7. Store all laboratory samples at 4º Celsius.
G. Biological Sampling 1. When collecting biological samples, scrape and rinse sediments from the grab into pre-cleaned containers. Save the rinse water* for infaunal sampling.
Protocols for Marine Environmental Monitoring 13 2. When sieving biological samples in the field:
§ Sieve each sediment sample, associated overlying water and rinse water through a 1.0 mm screen. Count, identify and record megafauna (e.g. large cnidarians, echinoderms and tube worms) then return them to the sea. Photograph specimens that need to be identified by taxonomists. Fix the remaining organisms in 10% buffered formalin. After 4 d, rinse over 0.5 mm screen and preserve in 70% isopropyl alcohol or ethyl alcohol. § Retain all coarse gravel and cobble less than 2.5 cm in diameter. § Remove epifauna adhering to rocks and other material that is greater than 2.5 cm diameter and include them in sieved sample. *Prior to use in sieving biological samples, rinse water must be filtered through a minimum 250 µm screen 3. For samples not sieved during the day they were obtained in the field use a 10% buffered formalin solution for preservation.
Reporting Submit monitoring data by filling out templates supplied by WLAP.
Protocols for Marine Environmental Monitoring 14
4. Checking the Quality of Sediment Samples
Physical and Chemical QA/ QC
Free Sulfides Take an additional sulphide measurement once every 20 samples, or once per batch if fewer than 20 samples are taken. See Section 7 for statistical approach to QA/QC.
Redox Potential
Take a triplicate measurement of Eh once every 20 samples, or once per batch if fewer than 20 samples are taken. See Section 7 for statistical approach to sulphide QA/QC.
TVS/ SGS/ Other Parameters Obtain additional sediment from 1 of every 20 sub-samples, or once per batch if fewer than 20 samples are taken, and have duplicate analyses of required parameters completed.
Biological QA/ QC There are 2 options for QA/ QC on biological samples:
1. Have certified facility staff submit QA samples to an expert contract taxonomist. The taxonomist’s lab must have its own QA/ QC program.
2. Have certified facility staff complete the taxonomy on site. For every 10 grab samples, they must take an additional grab sample, screen it, and split it into 2 samples. They count and identify one of the samples themselves, while submitting the other one to a recognized lab for the same procedure.
The results obtained by facility staff should match the lab results at a similarity level of at least 70%. Results from the contract lab must be reported to WLAP directly.
The facility staff must be certified by an educational institute recognized for expertise in taxonomy. Staff must be certified in taxonomic identification of benthic organisms to the family level.
Samples used by both the facility staff and the labs must be preserved and stored for a minimum of 5 years.
Reporting Submit data by filling out templates supplied by WLAP.
Protocols for Marine Environmental Monitoring 15
5. Calibrating the Sulphide Electrode
A sulphide electrode should be calibrated before each sampling session, and recalibrated every 3 - 4 hr during the session.
Materials § Orion model 290A meter and 9616BN silver/ sulphide electrode (Accumet and other brands are also acceptable.) § Electrolyte Solution: Ag/ Cl reference electrode filling solution Optimum Results A § Prepared solutions:
Solution Preparation frequency
Sulphide Anti-Oxidant Buffer (SAOB) every 3 to 4 hr
= -2 stock S solution 10,000 mM (10 M Na2S) every 5 d
= -3 standard S solution 1,000 mM (10 M Na2S): every 3 to 4 hr
= -4 standard S solution 100 mM (10 M Na2S) every 3 to 4 hr
= -5 standard S solution 10 mM (10 M Na2S) every 3 to 4 hr use only for a 3- point calibration of sediment samples with low sulphide concentrations
final calibration solutions immediately before calibration
Preparing Solutions
A. Sulphide Anti- Oxidant Buffer (SAOB)
1. Materials § 20.00 g NaOH (sodium hydroxide) § 17.9g EDTA § 8.75 g L-ascorbic acid § de-aerated DI or distilled water.
2. Procedures § In a 250 mL plastic screw top jar, mix the NaOH with the EDTA and dilute it to 250 mL with de-aerated DI or distilled water. This solution is stable for up to 7 d. (Larger or smaller volumes can be made up provided the ratios are maintained). § Do not add the L-ascorbic acid to EDTA/ NaOH solution until just before sample analysis, since the solution is stable for only 4 hr after adding the L-ascorbic acid. Store SAOB buffer in the dark at 4°C.
Protocols for Marine Environmental Monitoring 16 § Once SAOB is added to a sediment sample or sulphide standard, take measurements within 30 min.
= -2 B. Stock S solution 10,000 m M (10 M Na 2S)
1. Materials
§ 0.2402 g Na2S*9H20 (pre-weighed and stored under nitrogen) § de-aerated DI or distilled water
2. Procedures
§ In a well-ventilated area, add 0.2402 g Na 2S*9H20 to a volumetric flask and dilute to 100 mL using de-aerated DI or distilled water. § Store this stock solution in an airtight dark glass bottle at 4°C. Provided the head space is minimized, this stock solution is stable for up to 5 d.
= -3 C. Standard S solution 1,000 m M (10 M Na 2S)
1. Materials § stock solution § de-aerated DI or distilled water
2. Procedure § In a 100 mL volumetric flask, pipette 10 mL of the stock solution and dilute to 100 mL using de-aerated DI or distilled water. Store in an airtight dark glass bottle at 4°C.
= -4 D. Standard S solution 100 mM (10 M Na2S)
1. Materials § 1,000 µM solution § de-aerated DI or distilled water
2. Procedure § In a 100 mL volumetric flask, pipette 10 mL of the 1,000 µM solution and dilute to 100 mL using de-aerated DI or distilled water. Store in an airtight dark glass bottle at 4 °C.
= -5 E. Standard S solution 10 m M (10 M Na 2S) This solution is to be used only for a 3-point calibration for sediment samples with low sulphide concentrations.
1. Materials § 100 µM solution § de-aerated DI or distilled water
2. Procedure: § In a 100 mL volumetric flask, pipette 10 mL of 100 mM solution and dilute to 100 mL using de- aerated DI or distilled water. Store in an airtight dark glass bottle at 4°C.
Protocols for Marine Environmental Monitoring 17 F. Final Calibration Solutions
1. Materials § 25 mL SAOB buffer (containing L-ascorbic acid) § 25 mL Stock S = solution 10,000 mM § 25 mL Standard S= solution 1,000 mM § 25 mL Standard S= 100 mM
2. Procedures § Mix 25 mL of SAOB buffer and 25 mL Stock S= solution (10,000 mM) in a dark plastic 100 mL wide mouth bottle at 4°C. § Repeat preceding step for 1,000 mM solution and 100 mM (or 10 mM concentration as necessary). § Ensure that all calibration solutions are at the same temperature as the sediments being measured.
Procedures The sulphide electrode should be recalibrated between each set of samples or once every 3-4 hr, whichever is less.
A. Prepare the probe for calibration 1. Remove the cover from the electrode and connect the electrode to the meter.
2. Check the level of the probe’s filling solution, which should almost reach the filler hole. Add more solution if necessary. After filling a dry probe or topping up a low solution level, press down on the cap to wet the bottom O-ring and then tilt the container back to wet the top O-ring. Then add more solution to ensure the level almost reaches the filler hole.
B. Calibrate the probe for each standard solution Calibrating the probe against 3 standard solutions (a 3-point calibration) is recommended. Start with the least concentrated standard (10 µM or 100 mM) and progress to the most concentrated standard (1,000 mM or 10,000 µM). Select standards with concentration ranges that bracket the expected sulphide concentration of the samples
1. Press the mode button until the display indicates concentration mode.
2. Place the electrode in the lowest standard and until the meter reading stabilizes before beginning calibration process.
3. Press the second function button and then the calibrate bu tton. After a few seconds the lower field will read P1, indicating that the meter is ready for calibrating the first standard.
4. Press up arrow and 0.000 will appear.
5. Press up arrow again and decimal point will flash.
Protocols for Marine Environmental Monitoring 18 6. Press the up or down arrow keys to move the decimal place to the position you want (3 decimal places to the right for 100 mM and off the screen for 1,000 µM and 10,000 µM). Press YES when the decimal is in the correct position.
7. The first digit on the left will flash. Use the up or down arrow keys to change the digit, and press YES when the correct digit is displayed. To select 1,000 mM, press up or down arrow until first digit disappears, the press YES; for 10,000 µM press up or down arrow until 1 appears, then press YES. The next digit will then flash. Repeat the sequence for all digits until the readout displays the correct standard.
Once you have entered the first standard, the lower field will indicate the next standard to calibrate (e.g. P2 = second standard).
8. Rinse the electrode.
9. Repeat steps 4-7 until the 3 standards have been entered which completes the calibration.
C. Check the calibration 1. Press MEASURE. The meter will display the slope.
2. If the slope is between -27 and -33 record it on the data sheet. If the slope is outside this range, check the standards and calibrate the meter again.
3. Following calibration, rinse the electrode with DI or distilled water and blot it dry before measuring the first sample.
4. After taking the last sample in a series, rinse the electrode with distilled water and store in distilled water for short periods (up to one week). For longer periods, store it dry.
Protocols for Marine Environmental Monitoring 19
6. Standardizing the Redox Electrode
Materials § Orion model 290A meter and 9678BN combination redox electrode (Accumet or other brands will work as well). § electrolyte solution: Ag/ AgCl (silver chloride) reference electrode filling solution 900011 (correction factor at 20C is 204 mV). § redox standard (an Orion off-the-shelf standard triiodide/ iodide redox couple or other standards recommended by the manufacturer)
Procedures Standardization should occur every 4 hr or at the beginning and end of each transect:
1. Remove the cover from the electrode and connect the electrode to the meter.
2. Check the level of the filling solution. If necessary, add solution until it is at least one inch above the level of the solution being measured. After filling a previously dry probe or topping up the probe, push the cap and body together to leak some filling solution past the conical reference junction.
3. Place the electrode in the standard solution and wait until the reading stabilizes. Record the value of the standard and the meter reading at each standardization (for the triiodide/ iodide redox couple standard, the meter should read 220 mV – the potential of this standard solution).
4. The electrode is now ready for sampling. Insert it into the sample and record the mV reading after stabilization (when meter flashes READY or when drift is 3 mV or less over a 2 second period). Remove electrode and gently wipe off excess sediment prior to next measurement.
Note: This raw data is uncorrected. To correct the data, follow the procedure in Sediment Sampling: SECTION 3 F(5).
5. After use, rinse the electrode in DI or distilled water and store for short periods (a few weeks) in tap water. For longer periods, drain the electrode, rinse it in DI or distilled water, and store dry.
Protocols for Marine Environmental Monitoring 20
7. Performing Statistical Analyses
Preparations for Statistical Analyses These procedures must be followed before the data are analyzed using inferential statistics.
A. Validate sampling stations
1. Validate reference stations a. Confirm that there is more than one local reference station per facility. b. Confirm that reference stations are 0.5-2.0 km from the fa cility. c. Confirm that reference stations are at least 0.5 km apart. d. Confirm that mean depth at local reference stations is within 25% of that at facility stations. e. Confirm that mean % silt/ clay fraction at local reference stations is within 15% of that at facility stations.
2. Validate transects a. Confirm that transects were laid along prevailing currents. Current meter directions should be within 20o of transect directions.
B. Correct data entry errors
1. Identify dubious values (e.g. outliers) a. Draw graphs (e.g. box plots, scatter plots) b. Calculate summary statistics (e.g. x , M, SD, max, min)
2. Make corrections a. Contact data collectors for corrections b. Make necessary changes
C. Perform QA/QC for physical, chemical, & biological data
1. Check SGS data Confirm that 35% Relative Standard Difference has not been exceeded
= 2. Check S and Eh data Plot one variable against the other and look for outliers
3. Check TVS or TOC data Confirm that 20% Relative Percent Difference has not been exceeded
4. Check Cu and Zn data (to be added)
Protocols for Marine Environmental Monitoring 21 5. Check taxonomic data Confirm Similarity is at least 70%. See Appendix C for summary of tests.
Statistical Methods to Determine If Requirements Have Been Met These study designs and statistical tests are to be used to determine whether facilities are meeting chemical and biological requirements. See Appendix C for summary of tests.
A. Basic Study Designs
1. Existing facilities 2 basic designs are employed: a. MCI design – data are collected at facility stations and compared against data collected at reference stations b. Multiple Gradient design – data are collected at stations along multiple transects extending outward from the facility along prevailing currents, and related to distance from the facility
2. New facilities Beyond BACI design – data are collected at facility stations and reference stations, in both baseline and operational periods, to see if effect of facility/reference depends on baseline/operational. Ideally there are multiple sampling times in both periods.
B. Meeting Chemical Requirements
1. Existing facilities For stations located at or beyond the 30 m stations but within the tenure perimeter, first determine whether there has been a S= exceedance at any of these stations. Do this for each station by testing these h ypotheses using a 1-sample t-test:
HO : µ £ 1300 µM; HA : µ > 1300 µM (1-tailed)
HO : µ £ 6000 µM; HA : µ > 6000 µM (1-tailed) If there is evidence for an exceedance at a particular station, do analyses below to determine whether exceedance is due to fish farming or natural processes. a. MCI design For each station, perform Nested 1-way ANOVA to test this hypothesis:
HO : µF £ µR; HA: µF > µR (1-tailed) (F = facility, R = reference) If the facility station mean is significantly greater than the reference station mean, there is evidence that the exceedance is due to fish farming, and the requirement has not been met. Note that the above test is the same as a 2-sample t-test when design is balanced. Note also that this analysis may be superfluous if facility values are far above reference values. b. Multiple Gradient design Perform NLR, SLR, or MLR, depending on relationships. For example, perform NLR to test this hypothesis:
Protocols for Marine Environmental Monitoring 22
HO : ß1 ³ 0; HA : ß1 < 0 (1-tailed) If there is a significant non-linear decline outward from facility, then there is supporting evidence that the requirement has not been met. Alternatively, use a post hoc test (e.g. Tukey’s test), to make all possible pair-wise comparisons of distances, if more than one sample was taken at each distance. As above, a declining pattern provides supporting evidence that the requirement has not been met. Again, note that these analyses may be superfluous if facility values are far above reference values. For stations located at or beyond the perimeter of the tenure, do the above analyses without first testing for an exceedance.
2. New facilities For stations located at or beyond the 30 m stations but within the tenure perimeter, first determine whether there has been a S= exceedance at any of these stations using a 1-sample t-test as described above . If there is evidence for an exceedance at a particular station, do analysis below to determine whether exceedance is due to fish farming or natural processes. a. Beyond BACI design For each station, perform Asymmetric ANOVA to test the following hypotheses
HO: There is no interaction between facility/reference and baseline/operational; HA : there is an interaction (2-tailed). If there is a significant interaction, there is evidence that the exceedance is due to fish farming, and the requirement has not been met. Note that this analysis may be superfluous if facility values are far above reference or baseline values. For stations located at or beyond the perimeter of the tenure, do the above analyses without first testing for an exceedance.
C. Meeting Biological Requirements If a chemical requirement has not been met at a particular facility station, then biological analyses may be required for that station . Follow the methods below if biological analyses are required. Use taxon richness (total number of taxa) as the measure of diversity to be analyzed, and total number of individuals or total percent cover, as measures of total abundance to be analyzed. Data on biota from soft bottoms will be at the family, whereas those from hard substrata will be at class level.
1. Existing facilities a. MCI design For each station located at or beyond the 30 m stations but within the tenure perimeter, perform Nested 1-way ANOVA to test these hypotheses:
HO : µF ³ µR; HA: µF < µR (1-tailed) (F = facility, R = reference)
Protocols for Marine Environmental Monitoring 23 Note that the above test is the same as a 2-sample t-test when design is balanced. Report the results of these analyses in accordance with Section 6 (2)b of the Finfish Aquaculture Waste Control Regulation. For stations located at or beyond the perimeter of the tenure, do a 1-way Nested ANOVA to test these hypotheses:
HO : µF = µR; HA: µF ¹ µR (2-tailed) (F = facility, R = reference) If mean richness or mean total abundance at a particular station differs significantly from that at reference stations, then the requirement has not been met. b. Multiple Gradient design For each station located at or beyond the 30 m stations but within the tenure perimeter, perform NLR, SLR, or MLR, depending on relationships. For example, perform NLR to test these hypotheses:
HO : ß1 £ 0; HA : ß1 > 0 (1-tailed) Report the results of these analyses in accordance with Section 6 (2)b of the Finfish Aquaculture Waste Control Regulation and indicate if there is a significant non-linear increase in richness or abundance outward from the facility. Alternatively, use a post hoc test (e.g. Tukey’s test), to make all possible pair-wise comparisons of distances, if more than one sample was taken at each distance. Again, report the results of these analyses in accordance with Section 6 (2)b of the Finfish Aquaculture Waste Control Regulation and indicate any increases. For stations located at or beyond the perimeter of the tenure, do above analyses, except hypotheses should always be 2-tailed. If there are significant increases then the requirement has not been met.
2. New facilities a. Beyond BACI design For stations located at or beyond the 30 m stations but within the tenure perimeter, perform Asymmetric ANOVA to test these hypotheses for each station:
HO : There is no interaction between facility/reference and baseline/operational; HA: there is an interaction (2-tailed). Report the results of the analysis for both richness and abundance in accordance with Section 6 (2)b of the Finfish Aquaculture Waste Control Regulation and indicate if there are significant interactions. For stations located at or beyond the perimeter of the tenure, perform Asymmetric ANOVA to test the same hypotheses. If there is a significant interaction for either the richness analysis or the abundance analysis, then the requirement has not been met.
D. Analyses of Additional Variables In addition to the analyses described above, analyses of other physical, chemical, and biological variables may also be done in a weight-of-evidence approach to determine whether requirements have been met. Use contingency table analyses or ANOVA to determine whether gas bubbles, strong odours, black sediments, etc., are more common at facility stations than reference stations.
Protocols for Marine Environmental Monitoring 24 Use ANOVA and regression methods to analyze Eh, TVS, or TOC data, as was done above with S=, to test for differences and trends. Use contingency table analyses or ANOVA to determine if Beggiatoa or Capitella are more common at facility stations than reference stations. In addition to the analyses of means described above, analyses of standard deviations may prove useful in further defining the nature of fish farm effects. Use multivariate methods (e.g. MDS, ANOSIM) to determine whether community composition has been altered.
Statistical Power Analyses
A. Raising Statistical Power Due to the variable nature of the data and small sample sizes, statistical power will often be low. Power can be increased by doing the following:
1. Increasing N a. 1-sample t-tests Increase number of grabs or quadrats per facility station. b. MCI designs For ANOVAs, increase number of reference stations by “borrowing” reference stations from facilities in the same geographic region, or from facilities coast-wide. Alternatively, include nearby facilities belonging to the same company in same geographic region at similar stage of production cycle, along with their reference stations, in analyses. Note that including additional reference stations and facilities in analyses will have a much greater effect on power than increasing number of grabs or quadrats at each station. c. Multiple Gradient designs For regressions, increase number of stations sampled per transect, or increase number of grabs or quadrats per station. For post hoc tests, increase number of grabs or quadrats per station.
2. Increasing a For all tests, consider using the more precautionary a = 0.10 instead of the usual 0.05.
B. Estimating Desired Sample Sizes To determine sample sizes (numbers of transects, stations, grabs, quadrats) needed to achieve desired statistical power for future monitoring, power calculations must be done. 1 - b ³ 80% is recommended.
For both Multiple Gradient designs and MCI designs, power calculations will be based on EScrit values decided upon by the investigator. When doing any power calculations, consider using the more precautionary a = 0.10 instead of the usual 0.05, to raise power.
Protocols for Marine Environmental Monitoring 25
Appendix A: Design of Video Survey
Baseline Monitoring Parameter Sampling Units Sampling Locations Spatial Scale Min # Replicates
Class richness Transects Across e ntire tenure Length/width of Enough to ID biophysical and abundance tenure characteristics to 50 m of megafauna resolution
2 at each station* Reference stations At least 100 m long
Class richness Quadrats Entire tenure 1 x 1 m, with nine Enough to represent each and abundance 33 x 33 cm sections substratum type of macrofauna reference station As above 5 at each station
Notes
*1 transect runs perpendicular to shore .
Protocols for Marine Environmental Monitoring 26 Appendix B: Design of Sediment Sampling
Baseline Monitoring
Parameter Sampling Units Spatial Scale Sampling Locations Min # Grabs
S= Petite-Ponar, Ponar, Any size all stations 3 grabs per sediment type Smith-MacIntyre, or for each probable footprint. van Veen grab, etc. Minimum of 5 grabs if only 1 sediment type present.
Eh Petite-Ponar, Ponar, Any size all stations 3 grabs per sediment type Smith-MacIntyre, or for each probable footprint. van Veen grab, etc. Minimum of 5 grabs if only 1 sediment type present.
TVS or TOC Petite-Ponar, Ponar, Any size all stations 3 grabs per sediment type Smith-MacIntyre, or for each probable footprint. van Veen grab, etc. Minimum of 5 grabs if only 1 sediment type present.
SGS Petite-Ponar, Ponar, Any size all stations 3 grabs per sediment type Smith-MacIntyre, or for each probable footprint. van Veen grab, etc. Minimum of 5 grabs if only 1 sediment type present.
Cu or Zn Petite-Ponar, Ponar, Any size all stations 3 grabs per sediment type Smith-MacIntyre, or for each probable footprint. van Veen grab, etc. Minimum of 5 grabs if only 1 sediment type present.
Species richness Smith-MacIntyre, or 0.1 m2 all stations 3 grabs per sediment type and abundance of van Veen grab, etc. for each probable footprint. infauna and Minimum of 5 grabs if only epifauna 1 sediment type present.
Protocols for Marine Environmental Monitoring 27
Operational Monitoring Parameter Sampling Units Spatial Scale Sampling Location s Min # Samples
S= Petite-Ponar, Ponar, Any size All stations (see Notes: 3 grabs at all stations** Smith-MacIntyre, or van below) Veen grab, etc.
Eh Petite-Ponar, Ponar, Any size All stations (see Notes: 3 grabs at all stations** Smith-MacIntyre, or van below) Veen grab, etc.
TVS or TOC Petite-Ponar, Ponar, Any size Only at stations at 3 grabs at each station Smith-MacIntyre, or van perimeter of c.s. and located at perimeter of Veen grab, etc. reference stations c.s., and 3 at each reference station
SGS Petite-Ponar, Ponar, Any size Only at stations at 1 grab at each station Smith-MacIntyre, or van perimeter of c.s. and located at perimeter of Veen grab, etc. reference stations c.s., and 1 at each reference station
Cu or Zn Petite-Ponar, Ponar, Any size Only at stations at 3 grabs at each st ation Smith-MacIntyre, or van perimeter of c.s. and located at perimeter of Veen grab, etc. reference stations c.s., and 3 at each reference station
Family richness Smith-MacIntyre, or van 0.1 m2 All stations 5 grabs at each station, and abundance Veen grab, etc. except 3 at each of infauna and reference station epifauna*
*Biological sampling will only occur if a S= requirement has not been met.
**If the mean of the S= measurements from the 3 grabs exceeds 1300 µM, an additional 2 grabs must be obtained from that station for S= and Eh.
Notes: · Sampling station locations are: perimeter of containment structures; 30 m from zero metre station; perimeter of tenure; and reference stations. · Within tenure, have at least one transect for each dominant current direction or an alternate study design, provided extent and magnitude of effects are represented.
Protocols for Marine Environmental Monitoring 28 Appendix C: Statistical Procedures
Main statistical methods for different facility station locations, and differen t variables. Information for richness and abundance applies to both soft and hard bottom sites. Note that a 1-way Nested ANOVA is equivalent to a 2-sample t-test when design is balanced.
A. Existing Facilities
Free Sulphides Richness & Abundance
Wit hin Tenure Do 1-sample t-test (1-tailed), then if Do 1-way Nested ANOVA (1-tailed) necessary, do 1-way Nested ANOVA (1-tailed)
Tenure Perimeter Do 1-way Nested ANOVA (1-tailed) Do 1-way Nested ANOVA (2-tailed)
B. New Facilities
Free Sulphides Richness & Abundance
Within Tenure Do 1-sample t-test (1-tailed), then if Do Asymmetric ANOVA (2-tailed) necessary, do Asymmetric ANOVA (2- tailed)
Tenure Perimeter Do Asymmetric ANOVA (2-tailed) Do Asymmetric ANOVA (2-tailed)
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Protocols for Marine Environmental Monitoring 29