ROBERTS BANK TERMINAL 2
TECHNICAL DATA REPORT
Coastal Birds
Genetics of Western Sandpipers Using
the Fraser River Estuary during Northward Migration
Prepared for: Port Metro Vancouver 100 The Pointe, 999 Canada Place Vancouver, BC V6C 3T4 Prepared by: Hemmera Envirochem Inc. 18th Floor, 4730 Kingsway Burnaby, BC V5H 0C6 File: 302-042.02 December 2014
Port Metro Vancouver Hemmera RBT2 – WESA Genetics December 2014
Technical Report / Technical Data Report Disclaimer
The Canadian Environmental Assessment Agency determined the scope of the proposed Roberts Bank
Terminal 2 Project (RBT2 or the Project) and the scope of the assessment in the Final Environmental
Impact Statement Guidelines (EISG) issued January 7, 2014. The scope of the Project includes the
project components and physical activities to be considered in the environmental assessment. The scope
of the assessment includes the factors to be considered and the scope of those factors. The
Environmental Impact Statement (EIS) has been prepared in accordance with the scope of the Project
and the scope of the assessment specified in the EISG. For each component of the natural or human
environment considered in the EIS, the geographic scope of the assessment depends on the extent of
potential effects.
At the time supporting technical studies were initiated in 2011, with the objective of ensuring adequate
information would be available to inform the environmental assessment of the Project, neither the scope
of the Project nor the scope of the assessment had been determined.
Therefore, the scope of supporting studies may include physical activities that are not included in the
scope of the Project as determined by the Agency. Similarly, the scope of supporting studies may also
include spatial areas that are not expected to be affected by the Project.
This out-of-scope information is included in the Technical Report (TR)/Technical Data Report (TDR) for
each study, but may not be considered in the assessment of potential effects of the Project unless
relevant for understanding the context of those effects or to assessing potential cumulative effects.
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - i - December 2014
EXECUTIVE SUMMARY
Port Metro Vancouver (PMV) is assessing the potential to develop the Roberts Bank Terminal 2 Project
(RBT2 or the Project), a new three-berth marine terminal at Roberts Bank in Delta, B.C. The Project is
part of PMV’s Container Capacity Improvement Program (CCIP), a long-term strategy to deliver projects
to meet anticipated growth in demand for container capacity to 2030.
Hemmera has been retained by PMV to undertake environmental studies related to the Project. This
technical data report describes the results of the western sandpiper (Calidris mauri, hereafter referred to
as WESA) genetics study. WESA are small shorebirds that migrate long distances between breeding
grounds in western Alaska and easternmost Siberia, and wintering sites along the Pacific and Atlantic
coasts of the Americas (Wilson 1994). Roberts Bank and Sturgeon Bank, together with Boundary Bay,
are key stop-over points on the Pacific Flyway for millions of WESA on their spring migration.
Migratory species such as WESA have the potential to exhibit genetic structure meaning that individuals
within a species can be divided into groups or lineages based on their genetic make-up (COSEWIC
2011). Recent studies of non-breeding WESA populations in Mexico showed a modest amount of genetic
structure associated with migratory period and habitat type (Enriquez-Paredes et al. 2012). Presently, no
information exists on whether genetically distinct populations of WESA preferentially use Roberts Bank as
opposed to other parts of the Fraser River estuary (FRE) (Haig et al. 1997, Fernandez and Lank 2006).
The objective of this study was to determine if WESA visiting Roberts Bank are genetically differentiated
from those that visit other areas of the estuary (i.e., Sturgeon Bank or Boundary Bay). The study
consisted of three main components: 1) available literature and data review; 2) WESA sampling; and 3)
WESA genetic analysis.
Results show that little genetic structure was found among WESA using the Roberts Bank, Sturgeon
Bank and Boundary Bay sites within the FRE. There was a high level of variability within each site
(AMOVA), suggesting that many individuals from diverse lineages use the estuary. Levels of genetic
variability, expressed both as haplotype and nucleotide diversity, were highest at Roberts Bank,
suggesting that individuals from a large gene pool visit this site on their annual northward migration.
Genetic lineages of WESA do not appear to be segregating according to site within the FRE during
northward migration; therefore, WESA visiting Roberts Bank do not form a distinct genetic group.
Furthermore, haplotypes of individuals that visit the FRE during northward migration fall within the suite of
haplotypes and nucleotide diversity found in individuals overwintering in Mexico.
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TABLE OF CONTENTS
EXECUTIVE SUMMARY ............................................................................................................................... I
LIST OF ACRONYMS AND SYMBOLS ....................................................................................................... 1
1.0 INTRODUCTION .............................................................................................................................. 1
1.1 PROJECT BACKGROUND ........................................................................................................ 1
1.2 STUDY PURPOSE ................................................................................................................... 1
1.3 WESTERN SANDPIPER GENETICS STUDY OVERVIEW ............................................................... 1
2.0 REVIEW OF AVAILABLE LITERATURE AND DATA ................................................................... 3
2.1 MORPHOLOGICAL AND LIFE HISTORY DIFFERENTIATION IN WESTERN SANDPIPERS ................... 3
2.2 WESTERN SANDPIPERS POPULATION GENETICS ..................................................................... 3
2.3 GENETIC DIFFERENTIATION IN OTHER CALIDRIS SPECIES ........................................................ 4
2.4 SUMMARY ............................................................................................................................. 4
3.0 METHODS ....................................................................................................................................... 5
3.1 STUDY AREA ......................................................................................................................... 5
3.2 TEMPORAL SCOPE................................................................................................................. 7
3.3 STUDY METHODS .................................................................................................................. 7
3.3.1 Field Techniques and Observations ...................................................................... 7
3.3.2 DNA Extraction ....................................................................................................... 8
3.3.3 PCR and DNA Sequencing .................................................................................... 8
3.4 STATISTICAL, CALCULATION, AND MODELING APPROACHES ..................................................... 8
3.4.1 Sequence Editing and Alignment ........................................................................... 9
3.4.2 Diversity Indices ..................................................................................................... 9
3.4.3 Haplotype Frequency Histograms .......................................................................... 9
3.4.4 Phylogenetic Analysis ............................................................................................ 9
3.4.5 Analysis of Population Genetic Structure .............................................................. 9
3.5 DATA QUALITY AND INFORMATION MANAGEMENT .................................................................. 10
3.5.1 Sequence Editing and Comparison with Samples from Mexico .......................... 10
3.5.2 Mexico Haplotype Network and Frequency Histogram........................................ 10
4.0 RESULTS ...................................................................................................................................... 11
4.1 STUDY RESULTS ................................................................................................................. 11
4.1.1 Field Capture ........................................................................................................ 11
4.1.2 Data Analysis ....................................................................................................... 11
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4.1.3 Diversity and Haplotype Frequency ..................................................................... 11
4.1.4 Phylogenetic Information by Sampling Location .................................................. 11
4.1.5 Population Genetic Structure ............................................................................... 12
4.2 COMPARISONS WITH DATA FROM MEXICO ............................................................................. 14
5.0 DISCUSSION ................................................................................................................................. 17
5.1 DISCUSSION OF KEY FINDINGS ............................................................................................. 17
5.2 DATA GAPS AND LIMITATIONS .............................................................................................. 17
5.2.1 Missing Mexico Clade .......................................................................................... 17
5.2.2 Absence of Reference Site Samples ................................................................... 18
5.2.3 Absence of Data for Other Genetic Markers ........................................................ 18
6.0 CLOSURE ...................................................................................................................................... 19
7.0 REFERENCES ............................................................................................................................... 20
8.0 STATEMENT OF LIMITATIONS ................................................................................................... 23
List of Tables
Table 1 Western Sandpiper Genetics Study Components and Major Objectives ........................... 2
Table 2 Breakdown of Sample Sizes and Diversity Information for FRE Sites (Roberts Bank,
Sturgeon Bank and Boundary Bay) .................................................................................. 12
Table 3 FRE Population Pairwise FST Estimates ........................................................................... 15
List of Figures
Figure 1 Study Area and Western Sandpiper Capture Sites at Roberts Bank, Sturgeon Bank and
Boundary Bay ...................................................................................................................... 6
Figure 2 Haplotype Networks of the FRE Haplotypes .................................................................... 13
Figure 3 Haplotype Networks of the FRE with Mexico Haplotypes ................................................. 14
Figure 4 Haplotype Frequency Histogram of 607 bp of mtDNA Control Region Sequence Data
from the FRE (A) and Mexico (B) ...................................................................................... 16
List of Appendices
Appendix A Captured Western Sandpiper Morphometrics
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LIST OF ACRONYMS AND SYMBOLS
ASY After Second Year
AMOVA Analysis of Molecular Variance
bp Base Pairs
DNA Deoxyribonucleic Acid
Fst Fixation Index
FRE Fraser River estuary
h Haplotype Diversity
mtDNA Mitochondrial DNA
π Nucleotide Diversity
PCR Polymerase Chain Reaction
PMV Port Metro Vancouver
RAPD Random Amplified Polymorphic DNA
RBT2 Roberts Bank Terminal 2 Project
SY Second Year
WESA Western Sandpiper (Calidris mauri)
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1.0 INTRODUCTION
This section provides an overview of the study, including project background and study components and
major objectives.
1.1 PROJECT BACKGROUND
The Roberts Bank Terminal 2 Project (RBT2 or Project) is a proposed new three-berth marine terminal at
Roberts Bank in Delta, B.C. that could provide 2.4 million TEUs (twenty-foot equivalent unit containers) of
additional container capacity annually. The Project is part of Port Metro Vancouver’s Container Capacity
Improvement Program, a long-term strategy to deliver projects to meet anticipated growth in demand for
container capacity to 2030.
Hemmera was retained by PMV to undertake environmental studies related to the Project. This technical
data report describes the results of the western sandpiper (Calidris mauri, hereafter referred to as WESA)
genetics study.
1.2 STUDY PURPOSE
WESA are small shorebirds that migrate long distances between breeding grounds in western Alaska and
easternmost Siberia, and wintering sites along the Pacific and Atlantic coasts of the Americas (Wilson
1994). Roberts Bank and Sturgeon Bank, together with Boundary Bay, are key stop-over points on the
Pacific Flyway for millions of WESA on their spring migration.
Migratory species such as WESA have the potential to exhibit genetic structure meaning that individuals
within a species can be divided into groups or lineages based on their genetic make-up (COSEWIC
2011). Recent studies of genetic structure in non-breeding WESA populations in Mexico showed a
modest amount of genetic structure associated with migratory period and habitat type (Enriquez-Paredes
et al. 2012). Presently, no information exists on whether genetically distinct populations of WESA
preferentially use Roberts Bank as opposed to other parts of the Fraser River estuary (FRE) (Haig et al.
1997, Fernandez and Lank 2006). The objective of this study was to determine if WESA visiting Roberts
Bank are genetically differentiated from those that visit other areas of the estuary (i.e., Sturgeon Bank or
Boundary Bay).
1.3 WESTERN SANDPIPER GENETICS STUDY OVERVIEW
The WESA Genetics Study consisted of three main components: 1) available literature and data review;
2) WESA sampling; and 3) WESA genetic analysis. Study components, major objectives and a brief study
overview are provided in Table 1.
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Table 1 Western Sandpiper Genetics Study Components and Major Objectives
Study Component Major Objective Brief Overview
1) Available literature and data review
To review available information and state of knowledge regarding WESA genetics and to identify key data gaps and areas of uncertainty within the general RBT2 area.
Key findings and components identified from the literature and data review are summarised in this report. The literature and data review was completed before development of the 2012 field program.
2) Western sandpiper sampling
To capture WESA at three sites (Roberts Bank, Sturgeon Bank, and Boundary Bay) within the FRE and to obtain tissue samples for genetic analysis.
Mist-netting was conducted during the spring northward migratory period in April and May of 2012, and blood samples were collected for genetic analysis.
3) Western sandpiper genetic analysis
To identify genetic units of WESA using the FRE as a migratory stopover site.
DNA1 extraction and mitochondrial DNA
sequencing of blood samples was conducted on samples collected during mist-netting.
1 Deoxyribonucleic acid
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2.0 REVIEW OF AVAILABLE LITERATURE AND DATA
The following section provides an overview of available literature and data considered by the WESA
genetics study.
2.1 MORPHOLOGICAL AND LIFE HISTORY DIFFERENTIATION IN WESTERN SANDPIPERS
WESA are small shorebirds that migrate long distances between breeding grounds in western Alaska and
easternmost Siberia, and wintering sites along the Pacific and Atlantic coasts of the Americas (Wilson
1994). Tidal mudflats, salt marshes, and sand beaches on the Pacific Coast (from Washington State to
Peru), the Atlantic Coast (from New Jersey to Surinam) and the Caribbean serve as overwintering sites
(Burger 1984, Wilson 1994). The majority of the population spends the non-breeding period in tropical
areas. WESA show very structured morphological variation across their non-breeding range with
individuals at southern wintering sites having longer wings and bills than those wintering further north
(Nebel 2005, O'Hara et al. 2006). In addition, latitudinal segregation by sex is observed, with males
wintering farther north than females (Fernández and Lank 2006). Life history strategies also differ across
the non-breeding range. Juvenile birds that spend the non-breeding season in Mexico return to the
breeding grounds in their first year of life, whereas those wintering in Panama are more likely to delay
reproduction until the following year (Fernández et al. 2004, O'Hara et al. 2005). It is unknown whether
this structured morphological variation is reflected in genetic differentiation (Lank and Nebel 2006).
2.2 WESTERN SANDPIPERS POPULATION GENETICS
At present, little is known regarding the population genetics of WESA. The first study to investigate WESA
population genetics employed the Random Amplified Polymorphic DNA (RAPD) technique to compare
genetic marker frequencies across one breeding site in Alaska and three wintering and/or migratory
stopover sites (Haig et al. 1997). Of the latter three sites, one site was located on the Pacific Coast in
California, and the remaining two were located on the Atlantic Coast in South Carolina. Although sample
sizes were low (5 to 10 birds per site), results indicated differences in RAPD marker frequencies across
sites (Haig et al. 1997). More recently, analysis of WESA mitochondrial DNA (mtDNA) from birds
overwintering in northern Mexico revealed the presence of two distinct groups of haplotypes (i.e., a group
of genes that is inherited together by an organism from a single parent), termed lineages ‘A’ and ‘B’.
Haplotypes of lineage A were more evenly distributed spatially and temporally, across all habitat types
and the overwintering vs. migratory periods, respectively. In contrast, haplotypes of lineage B were found
only during winter in mangrove and cattail marsh habitats, and resulted in low, but statistically significant
genetic structuring across sites and time-periods (Enríquez-Paredes et al. 2012).Whether individuals
from the two different lineages originated from different breeding areas could not be determined
(Enríquez-Paredes et al. 2012). MtDNA sequencing of additional samples from various sites along the
Pacific Coast and one site on the Atlantic Coast of Mexico also indicates haplotype structure (Enríquez
and Fernández 2010).
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2.3 GENETIC DIFFERENTIATION IN OTHER CALIDRIS SPECIES
The degree of population structure evident in a particular species varies considerably across other
calidrid sandpipers (Haig et al. 1997). Dunlin (Calidris alpina), for instance, exhibit marked genetic
differentiation across their circumpolar breeding range, and among wintering and migratory stopover sites
(Wenink et al. 1993, 1994; Wennerberg et al. 1999; Wennerberg 2001). MtDNA analyses revealed the
presence of five distinct lineages in dunlin with statistically significant differences in the presence and
frequencies of haplotype groups among geographic locations (Wenink et al. 1993, Wenink et al 1996;
Wennerberg et al. 1999, Wennerberg 2001). The mtDNA lineages correspond to five subspecies
previously defined on the basis of morphology and molt patterns. Further, there is some correlation
between morphological characteristics and haplotype (Wenink et al. 1993, Wenink and Baker 1996,
Wenink et al. 1996); however, the other morphologically defined subspecies remain unaccounted for by
either mtDNA or other analyses (Wenink et al. 1996, Marthinsen et al. 2007).
Statistically significant population differentiation has also been detected in pectoral and semipalmated
sandpipers (Calidris melanotos and Calidris pusilla, respectively; Haig et al. 1997). In rock sandpipers
(Calidris ptilocnemis), two morphologically defined subspecies showed clear phylogenetic differentiation,
but two others could not be distinguished genetically (Pruett and Winker 2005).
Several calidrid sandpiper species show extremely low levels of population structure with weak or
no statistically significant differentiation (Wennerberg et al. 2002; Bühler and Baker 2005; Rönkä et al.
2008, 2012). While Temminck’s stints (Calidris temminckii) and red knots (Calidris canutus) have low
levels of differentiation, white-rumped sandpipers (Calidris fuscicollis) do not show statistically significant
genetic differentiation despite morphological differences between northern and southern breeding
populations (Wennerberg et al. 2002; Bühler and Baker 2005; Rönkä et al. 2008, 2012).
2.4 SUMMARY
Calidrid sandpipers vary widely in their degree of genetic differentiation. WESA show morphological and
life history differences across their non-breeding range, but the degree to which phenotypic differentiation
is reflected in population genetic differentiation is presently unknown. This uncertainty identified the need
to conduct a WESA genetics study to get a better understanding of WESA populations in the RBT2
study area.
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3.0 METHODS
Descriptions of the study spatial and temporal scopes, and field sampling and sample analysis methods
are provided below.
3.1 STUDY AREA
The area of interest for the WESA genetic study is the FRE (Figure 1). For the purpose of this analysis,
the estuary was separated into three sites: Roberts Bank, Sturgeon Bank and Boundary Bay. These
locations were chosen because they collectively form one of the key stop-over sites on the Pacific Flyway
for millions of WESA during northward and southward migration (Catherine Berris Associates Inc. 2010).
The objective of this study was to determine if WESA visiting Roberts Bank could be genetically
differentiated from those that visit other areas of the estuary; therefore, sampling was limited to the FRE.
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Figure 1 Study Area and Western Sandpiper Capture Sites at Roberts Bank, Sturgeon Bank and Boundary Bay
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3.2 TEMPORAL SCOPE
WESA genetics studies were intended to assess possible genetic structures of northward migrating
WESA in the FRE. Results described in this report represent genetic variation found during the northward
migration in the spring of 2012 and may be an indication of patterns of genetic differentiation during
subsequent northward WESA migrations. Mist-netting to capture WESA occurred on 10 days over the
three-week northward migratory period from April 19 to May 6, 2012. Netting was conducted on rising and
falling tides corresponding to peak high-tide events under appropriate weather conditions (i.e., under dry
conditions with winds below 30 km/h [<Beaufort 5]). Some netting was conducted during the night due to
tide-height constraints.
3.3 STUDY METHODS
Descriptions of the field sampling and sample analysis methods are provided below.
3.3.1 Field Techniques and Observations
Mist net capture of WESA individuals occurred during the spring 2012 field season, which coincided with
the peak migratory period of WESA. Blood samples from WESA individuals were collected in conjunction
with feather samples for the study of WESA migratory connectivity (Hemmera 2014).
3.3.1.1 Field Sampling
Bird handling and animal husbandry skills were required in this study. All field crew personnel were
qualified and permitted for shorebird handling, and specialists were available throughout the sampling
period to consult on bird handling. Locations of mist nets at each of the sites are depicted in Figure 1.
The timing of WESA migration is stratified by age and sex (Butler et al. 1987, Mathot and Elner 2004);
therefore, three sampling periods were established within the WESA peak migration to capture this
variation. During each sampling period, a per-site quota was set for WESA capture so that an
approximately equal number of individuals were captured throughout the migration
(22±4 individuals/site/period). This system was developed to improve chances that a representative
sample of WESA sex and age classes was collected at each of the three sites during the
migratory period.
Mist nets were set up approximately one hour before the peak high tide on a rising tide or one hour before
exposure of intertidal mudflats on a falling tide. Playbacks of social calls were used during night captures
to draw WESA towards the nets. Nets were monitored continuously and taken down after approximately
three hours or when the daily sample size objectives were reached.
Protocol for bird capture and tissue sampling closely followed that of Enriquez-Paredes et al. (2012).
Upon capture, standard morphological measurements of each bird were taken, including bill length for
sex assignment and plumage coloration for age estimate. Blood samples (30 to 60 µL) for DNA extraction
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were drawn from the brachial artery on the wing. Blood sampling protocols are thought to minimally affect
the health of individual birds (Hoysak and Weatherhead 1991). Blood samples were deposited in
‘Queens’ storage buffer (see Seutin et al. 1990) and stored on ice for 4-6 hours until they were brought
back to the laboratory. Samples were stored in the laboratory refrigerator at approximately 3 °C.
3.3.2 DNA Extraction
Samples were sent to Genome Quebec for preparation and sequencing. In total 37 samples from
Boundary Bay, 30 Samples from Roberts Bank and 33 samples from Sturgeon Bank were sent to
Genome Quebec. Three samples from Sturgeon Bank and 7 samples from Boundary Bay were sent to
Genome Quebec as trial samples before the remaining 30 samples from each population were sent for
analysis. To optimise sampling over the entire migratory period, samples were selected from each
sampling location so that there were an approximately equal number of individuals from each sampling
location throughout the duration of the migratory period, with the exception of Roberts Bank, where no
individuals were captured during the early migratory period.
DNA extraction was performed at Genome Quebec using Qiagen DNeasy Blood & Tissue Kit (Qiagen
cat: 69504). The protocol entitled Purification of Total DNA from Animal Blood or Cells (Spin-Column
Protocol) with Nucleated Erythrocytes was used. One modification was made by Genome Quebec upon
receipt of the samples: In the protocol step 1b, 220 µL of the sample was gathered for analysis, taking
into consideration that the storage buffer that the blood samples were delivered in would replace the
Phosphate Buffered Solution buffer in the extraction protocol.
3.3.3 PCR and DNA Sequencing
The polymerase chain reaction (PCR) was conducted on a MasterCycler ProS - Vapo protect from
Eppendorf. Forward and reverse primer sequences were derived from Enriquez-Paredes et al. (2012):
CmCRF-(5’-CCCCCATACTACATACCATC-3’) and CmCRR – (5’-GTCCCACAAGCATTCATT-3’).
Thermal cycling conditions were as follows: initial denature step at 96 °C for 1min, followed by 33 cycles
of 96 °C for 10 seconds, 52 °C for 5 seconds and 72 °C for 1 second with a final denature step of 72 °C
for 30 seconds. Each PCR reaction was created to a total volume of approximately 25 µl per reaction and
included 19 µl ddH2O, 5 µl 5x PCR Buffer, 0.5 µl of dNTP, 0.1 µl TAQ KAPA2G Fast HotStart 5U/µl, 0.1 µl
CmCRF and 0.1 µl CmCRR primers, and 0.1 µl of DNA per reaction. DNA was sequenced on a 3730xl
DNA Analyser from Applied Biosystems.
3.4 STATISTICAL, CALCULATION, AND MODELING APPROACHES
Descriptions of approaches taken for statistical, calculation, and modelling methods are provided below.
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3.4.1 Sequence Editing and Alignment
Sequences were edited using Four Peaks v.1.7.2 (Griekspoor and Groothuis 2005), and Mesquite
(Maddison and Maddison 2011), and aligned using Clustal W multiple alignment software (Larkin et al.
2007).
3.4.2 Diversity Indices
Molecular diversity indices were calculated using Arlequin (Excoffier et al. 2005). Indices included mean
haplotype diversity (h; measures the uniqueness of a given haplotype within a sample) and mean
nucleotide diversity (π; average number of nucleotide differences per site measured between any
two individuals). Genetic distance was calculated using Kimura 2 Parameter distance, which uses
information about DNA transitions and transversions to calculate a distance metric (Kimura 1980).
All diversity data and subsequent analyses were conducted first on the full set of samples, including all
individuals, then on a subset of the data which included either, only males, only females or only juveniles.
3.4.3 Haplotype Frequency Histograms
A histogram of the haplotype frequency (relative number of individuals per haplotype) was constructed in
PASW Statistics 18.
3.4.4 Phylogenetic Analysis
Phylogenetic haplotype networks, which outline linkages among mitochondrial variants within the sample,
were constructed in Network 4.6.1.1, using the Median Joining and Maximum Parsimony algorithms
(Bandelt et al. 1999, Polzin and Daneshmand 2003).
3.4.5 Analysis of Population Genetic Structure
Analysis of population genetic structure was conducted using Arlequin (Excoffier et al. 2005). Sequences
were analysed for pairwise population differentiation among sites (Roberts Bank, Sturgeon Bank, and
Boundary Bay) and analysis of molecular variance (AMOVA) was used to determine overall partitioning of
genetic variability within sites and among sites throughout the FRE.
A total FRE-wide sample was analysed, which included all sequences collected as part of this study.
Subsequent analyses of population genetic structure were conducted on subsets of the data that included
males only, females only, ASY (after second year) only and SY (second year) only to determine if genetic
structure could be detected in any of these groups independently. Finally, an AMOVA was used to
investigate whether genetic variability could be partitioned among males and females across the FRE.
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3.5 DATA QUALITY AND INFORMATION MANAGEMENT
All data was stored within the Hemmera Geomatics Database system.
3.5.1 Sequence Editing and Comparison with Samples from Mexico
FRE DNA sequences were edited in Four Peaks (Griekspoor and Groothuis 2005). Both forward and
reverse sequence data was utilised for each individual. Forward sequence reads tended to degrade
around 465 base pairs (bp). At this point the upstream (5’) end of the reverse compliment of the reverse
sequence read and the 5’ end of the forward sequence were concatenated into the full sequence (length
607 bp) ensuring that sequences of the highest quality were used in the analysis.
Sequences collected by Enriquez-Paredes et al. (2012) were downloaded from the Genbank database
(Accession Numbers: JF720884 - JF720924) and aligned with FRE data using Clustal W multiple
alignment software (Larkin et al. 2007).
3.5.2 Mexico Haplotype Network and Frequency Histogram
Mexican sequences were used as reference samples for the analysis of FRE WESA genetic data. Upon
alignment of FRE sequences with sequences from Enriquez-Paredes et al. (2012), a haplotype network
was constructed using NETWORK 4.6.1.1, using the Median Joining and Maximum Parsimony algorithms
(Bandelt et al. 1999, Polzin and Daneshmand 2003). In order to compare haplotype frequencies and the
relative diversity in the two datasets, a haplotype frequency histogram was constructed in PASW
Statistics 18, using all available WESA sequence data from Enriquez-Paredes et al. (2012) and from this
study of the FRE.
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4.0 RESULTS
This section presents the main findings of the study and provides a comparison of data collected from a
similar study conducted in Mexico.
4.1 STUDY RESULTS
This section discusses the results of the WESA genetics study conducted at the Fraser River estuary.
4.1.1 Field Capture
Overall, 199 WESA individuals were caught, measured, and sampled for blood analysis at Roberts Bank,
Sturgeon Bank and Boundary Bay (Figure 1). Of these, 36 individuals were captured at Roberts Bank,
67 at Sturgeon Bank, and 95 at Boundary Bay. Sampling was approximately evenly spread between the
April 19th and May 6
th 2012 except in the case of Roberts Bank where no individuals were captured on
April 20th or April 22
nd. The first individuals captured from Roberts Bank were caught on April 24 2012.
Males and females were found at all three sites, as were ASY and SY individuals (Appendix A and
Table 2).
4.1.2 Data Analysis
Eighty-eight WESA individuals were sequenced for a 607 bp long section of the mitochondrial control
region (Enriquez-Paredes et al. 2012). Eighteen birds of 30 individuals were reliably sequenced from
Roberts Bank, 37 of 37 individuals were reliably sequenced from Sturgeon Bank, and 33 of 33 individuals
were reliably sequenced from Boundary Bay. .
4.1.3 Diversity and Haplotype Frequency
In total, 34 unique haplotypes (i.e., maternal lineages) were sequenced in the three FRE sites. Five
haplotypes were found either at two or at three sites and the remaining haplotypes were found only at
Roberts Bank (6), Sturgeon Bank (10), or Boundary Bay (13). Haplotype diversity by site (proportion of
the total haplotypes) ranged from 0.59 to 0.79, and was highest at Roberts Bank. Nucleotide diversity
(number of segregating nucleotide sites) ranged from 0.0016 to 0.0018 and was also highest at Roberts
Bank (Table 2).
4.1.4 Phylogenetic Information by Sampling Location
Of the 88 haplotypes sampled in the FRE, the WESA haplotype network showed one high frequency
haplotype that consisted of sequences from over 50% of the individuals sampled (n = 52). The central
common haplotype was found in relatively equal frequencies in all three sites. This common haplotype is
homologous (in part: 607 bp) to the central haplotype in lineage A from Enriquez-Paredes et al. (2012)
(Figures 2 and 3).
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4.1.5 Population Genetic Structure
No evidence of population genetic structure was detected when all sequences were compared among
Roberts Bank, Sturgeon Bank, and Boundary Bay sites (Table 2). AMOVA results indicated a large but
statistically non-significant proportion of the total genetic variation could be explained within sites (100%,
P = 0.71). The among-site variability explained a lower, also not statistically significant, proportion of the
total genetic variability (42%, P = 0.71).
When the dataset was broken down into sex and age classes, there was no significant genetic
differentiation among sites in either gender group or age class group. Additionally, approximately 100% of
the total overall variation could be explained by within-population variation in AMOVA analyses of these
groupings. Finally, there was no statistically significant genetic variability between males and females
sampled throughout the FRE.
Table 2 Breakdown of Sample Sizes and Diversity Information for FRE Sites (Roberts Bank, Sturgeon Bank and Boundary Bay)
Site N Gender Age
1 Haplotype Nucleotide
Male Female Unknown SY
ASY
Diversity2
Diversity3
Roberts Bank 18 12 6 0 10 8 0.70 0.002
Boundary Bay 36 18 18 0 14 22 0.66 0.002
Sturgeon Bank 33 14 17 2 4 29 0.59 0.002
Notes: 1
Age: SY = second year; ASY = after second year. 2
Haplotype diversity is the relative proportion of unique haplotypes in the sample. 3
Nucleotide diversity is the relative proportion of nucleotide substitutions in each sequence at each site. Nucleotide diversity values are rounded to the nearest 1/1000
th.
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 13 - December 2014
Figure 2 Haplotype Networks of the FRE Haplotypes
Note: The size of each circle represents the frequency of each haplotype, such that larger circles are equal to
higher frequency haplotypes. Haplotype color represents sampling location, where red haplotypes indicate Boundary Bay, yellow haplotypes indicate Roberts Bank, black haplotypes indicate Sturgeon Bank. The lines connecting haplotypes indicate single substitution differences. Open circles represent haplotypes that are missing in the dataset.
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 14 - December 2014
Figure 3 Haplotype Networks of the FRE with Mexico Haplotypes
Note: Each circle represents a unique haplotype. The size of each circle represents the frequency of each
haplotype, such that larger circles are equal to higher frequency haplotypes. Haplotype color represents sampling location, where red haplotypes indicate Boundary Bay, yellow haplotypes indicate Roberts Bank, black haplotypes indicate Sturgeon Bank, and blue haplotypes indicated Mexico. The lines connecting haplotypes indicate single substitution differences. Open circles represent haplotypes that are missing in the dataset.
4.2 COMPARISONS WITH DATA FROM MEXICO
Forty-one unique haplotypes (i.e., those found in the 2012 study) from Mexico (Enriquez-Paredes et al.
2012) were downloaded from Genbank. Downloaded sequences contained 685 bp from the mitochondrial
control region.
More individuals were sampled in Mexico (163) than on the FRE (88), and the number of unique
haplotypes varied accordingly (Mexico 41, FRE 34). Nucleotide diversity was on the same order of
magnitude within the two studies, with a mean value of 0.0017 in the FRE and a mean value of 0.005 in
the Mexico sample.
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 15 - December 2014
After trimming all sequences from Enriquez-Paredes et al. (2012) to 607 bp, a haplotype frequency
histogram depicting both the FRE haplotypes and the haplotypes from Mexico showed that the most
common haplotype was identical between the two datasets as were the second and third most common
haplotypes; however, only two additional haplotypes were identical between the two datasets. Aside from
the shared haplotypes, each dataset also had many unique or rare haplotypes (Figure 4). In a haplotype
network consisting of Mexico samples (trimmed to 607 bp) and FRE samples, a second lineage that was
present at the Mexican site was missing in the FRE (Figure 3). This lineage is associated with variation in
ecological conditions on the wintering grounds in Mexico (see Enriquez Paredes et al. 2012). Levels of
population structure measured as FST were smaller within the FRE than among the Mexican sites
(Enriquez-Paredes et al. 2012) (Table 3). None of the possible pairwise population genetic differentiation
estimates on the FRE were statistically significantly different from zero.
Table 3 FRE Population Pairwise FST Estimates
Site Sturgeon Bank Boundary Bay
Sturgeon Bank
Boundary Bay 0.004
Roberts Bank -0.005 -0.004
Note: Pairwise FST estimates were not significantly different from zero.
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 16 - December 2014
Figure 4 Haplotype Frequency Histogram of 607 bp of mtDNA Control Region Sequence Data from the FRE (A) and Mexico (B)
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 17 - December 2014
5.0 DISCUSSION
A discussion of the major results arising from the WESA genetics study and data gaps are provided
below.
5.1 DISCUSSION OF KEY FINDINGS
Little genetic structure was found among Roberts Bank, Sturgeon Bank and Boundary Bay sites, within
the FRE. There was a high level of variability within each site (AMOVA), suggesting that many individuals
from diverse lineages use the estuary. Levels of genetic variability, expressed both as haplotype diversity
and nucleotide diversity were highest at Roberts Bank, suggesting that individuals from a large gene pool
visit this site on their annual northward migration.
Results of this study are consistent with other studies on within-region analysis of shorebird genetic
differentiation (Haig et al. 1997, Álvarez-Sánchez 2011, Enriquez-Paredes et al. 2012). Furthermore,
these results are not unusual, as studies of other calidrid shorebirds also detected low levels of
genetic differentiation among populations (Wennerberg et al. 2002; Bühler and Baker 2005; Rönkä et al.
2008, 2012).
The key findings from this study include:
High genetic diversity within sites, but low genetic differentiation among sites within the FRE,
suggesting that no site is genetically distinct within the estuary.
Highest levels of genetic variability at Roberts Bank, suggesting that a large gene pool of
individuals visit this site during their annual northward migration.
Genetic samples from the FRE had the same level of variability as samples from wintering
grounds in Mexico (Enriquez-Paredes et al. 2012).
One genetically distinct clade in Mexico (lineage B) was absent from the samples collected within
the FRE, suggesting that potentially ecologically differentiated populations in Mexico (i.e., the
bulrush population) (Enriquez-Paredes et al. 2012) did not use the FRE during their northward
migration in 2012.
5.2 DATA GAPS AND LIMITATIONS
This section provides an overview of the data gaps and limitations encountered during the WESA
genetics study.
5.2.1 Missing Mexico Clade
In all haplotypes from the FRE, the substitutions that constitute lineage B in Mexico were not found. This
implies that lineage B likely does not visit the FRE during its migration northward to the breeding grounds.
Other possible explanations include temporal shifts in migration for the missing Mexican clade such that it
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 18 - December 2014
passed through the FRE before or after this study captured birds; or it is possible that the missing Mexico
clade exists in very low frequencies relative to the entire population and thus our sampling effort missed
sampling individuals in lineage B purely by chance. The approximately 75 bp that were cleaved from
the 3’ end of the sequences from Mexico did not contain the substitutions that formed lineage B, which
suggests that this lineage may be truly missing from the FRE and not an artifact of sequence alignment
or sampling.
5.2.2 Absence of Reference Site Samples
At present, the only available published data on WESA mtDNA comes from the Enriquez-Paredes et al.
(2012) study in Mexico. Data from other parts of the range are lacking; therefore, it is possible that
stopover sites outside of the FRE region are genetically different.
5.2.3 Absence of Data for Other Genetic Markers
The current study lacks data from multiple nuclear and/or mitochondrial loci. The use of additional loci
would give a broader representation of the patterns of population genetic variability in WESA; however,
Enriquez-Parades et al. (2012) find that the mitochondrial Cytochrome Oxidase 1 locus (used in this
study) was sufficient to detect genetic differentiation in wintering Mexican populations.
Despite the above mentioned limitations, the major objectives as outlined in section one have effectively
been achieved.
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 19 - December 2014
6.0 CLOSURE
Major authors and reviewers of this technical data report are listed below, along with their signatures.
Report prepared by: Report peer reviewed by: Hemmera Envirochem Inc. Hemmera Envirochem Inc.
Carson Keever, PhD James Rourke, M.Sc. R.P.Bio. Technical Specialist Coastal Birds Discipline Lead Report peer reviewed by: Simon Fraser University
Ron Ydenberg, PhD Professor, Centre for Wildlife Ecology Director
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 20 - December 2014
7.0 REFERENCES
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occidental en México. M.Sc., Universidad Autónoma de Baja California, Ensenada, Baja
California.
Bandelt, H. J., P. Forster, and A. Röhl. 1999. Median-joining networks for inferring intraspecific
phylogenies. Molecular Biology and Evolution 16:37-48.
Bühler, D. M., and A. J. Baker. 2005. Population divergence times and historical demography in red knots
and dunlins. The Condor 107:497-513.
Burger, J. 1984. Shorebirds as marine animals. Pages 83-123 in Shorebirds - breeding behavior and
populations, vol. 5 (J. Burger, B. L. Olla, Ed.). Plenum Publishing Corporation, New York.
Butler, R. W., G. W. Kaiser, and G. E. J. Smith. 1987. Migration chronology, length of stay, sex ratio, and
weight of western sandpipers, (Calidris mauri) on the south coast of British Columbia. Journal of
Field Ornithology 58:103-111.
Catherine Berris Associates Inc. 2010. Roberts Bank and Sturgeon Bank Reach Overview: Phase 2.
Burrard Inlet Environmental Action Program.
Committee on the Status of Endangered Wildlife in Canada (COSEWIC). 2011. Designatable units for
caribou (Rangifer tarandus) in Canada. Ottawa. 88 pp.
Enríquez-Paredes, L. M., C. Vilanova, and G. Fernández. 2012. Genetic diversity and population
structure of wintering western sandpipers from the Sinaloa coast, Mexico. Journal of Field
Ornithology 83:85-93.
Enríquez, L., and G. Fernández. 2010. Genetic identity and population structure of nonbreeding western
sandpiper in México. Page 26 in COS/AOU/SCO 2010 Joint Meeting San Diego, California, USA.
Excoffier L., G. Laval, and S. Schneider. 2005. Arlequin (Version 3.0): an integrated software package for
population genetics data analysis. Evolutionary Bioinformatics Online 1: 47–50.
Fernández, G., and D. B. Lank. 2006. Sex, age and body size distributions of western sandpipers during
the nonbreeding season with respect to local habitat. The Condor 108: 547-557.
Fernández, G., P. D. O'Hara, and D. B. Lank. 2004. Tropical and subtropical western sandpipers (Calidris
mauri) differ in life history strategies. Ornitologia Neotropical 15:385-394.
Griekspoor, A. and Groothuis, T. 2005. 4 Peaks. Version 1.7.
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Haig, S. M., C. L. GrattoTrevor, T. D. Mullins, and M. A. Colwell. 1997. Population identification of western
hemisphere shorebirds throughout the annual cycle. Molecular Ecology 6:413-427.
Hemmera. 2014. Roberts Bank Terminal 2 technical data report: Migratory connectivity of western
sandpipers using the Fraser River estuary. Prepared for Port Metro Vancouver. Vancouver, B.C.
Available at: http://www.robertsbankterminal2.com/.
Hoysak, D., and P. Weatherhead. 1991. Sampling blood from birds: a technique and an assessment of its
effects. The Condor 93(3): 746-752.
Kimura, M. 1980. A simple method for estimating evolutionary rates of the base substitutions through
comparative studies of nucleotide sequences. Journal of Molecular Evolution 16: 111-120
Lank, D. B., and S. Nebel. 2006. Cross-cutting research on a flyway scale – beyond monitoring. Pages
107-112 in Waterbirds around the world (G. C. Boere, C. A. Galbraith, and D. A. Stroud, Eds.).
The Stationery Office, Edinburgh, UK.
Larkin, M. A., G. Blackshields, N. P. Brown, R. Chenna, P. A. McGettigan, H. McWilliam, F. Valentin, I. M.
Wallace, A. Wilm, R. Lopez, J. D. Thompson, T. J. Gibson, and D. G. Higgins. 2007. ClustalW
and ClustalX version 2. Bioinformatics 23(21): 2947-2948.
Maddison, W. P., and D.R. Maddison. 2011. Mesquite: a modular system for evolutionary analysis.
Version 2.75. Available at http://mesquiteproject.org.
Marthinsen, G., L. Wennerberg, and J. T. Lifjeld. 2007. Phylogeography and subspecies taxonomy of
dunlins (Calidris alpina) in western Palearctic analysed by DNA microsatellites and amplified
fragment length polymorphism markers. Biological Journal of the Linnean Society 92:713-726.
Mathot, K. J., and R. W. Elner. 2004. Evidence for sexual partitioning of foraging mode in western
sandpipers (Calidris mauri) during migration. Canadian Journal of Zoology 82(7):1035-1042.
Nebel, S. 2005. Latitudinal clines in bill length and sex ratio in a migratory shorebird: a case of resource
partitioning? Acta Oecologica-International Journal of Ecology 28:33-38.
O'Hara, P. D., G. Fernández, F. Becerril, H. de la Cueva, and D. B. Lank. 2005. Life history varies with
migratory distance in western sandpipers (Calidris maur)i. Journal of Avian Biology 36:191-202.
O'Hara, P. D., G. Fernández, B. Haase, H. de la Cueva, and D. B. Lank. 2006. Differential migration in
western sandpipers with respect to body size and wing length. The Condor 108:225-232.
Pruett, C. L., and K. Winker. 2005. Biological impacts of climatic change on a Beringian endemic: Cryptic
refugia in the establishment and differentiation of the rock sandpiper (Calidris ptilocnemis).
Climatic Change 68:219-240.
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Polzin, T., and S. V. Daneshmand. 2003. On Steiner trees and minimum spanning trees in hypergraphs.
Operations Research Letters 31: 12-20.
Rönkä, A., L. Kvist, J. Karvonen, K. Koivula, V. M. Pakanen, D. Schamel, and D. M. Tracy. 2008.
Population genetic structure in the Temminck's stint Calidris temminckii, with an emphasis on
Fennoscandian populations. Conservation Genetics 9:29-37.
Rönkä, N., L. Kvist, V. M. Pakanen, A. Ronka, V. Degtyaryev, P. Tomkovich, D. Tracy, and K. Koivula.
2012. Phylogeography of the Temminck's stint (Calidris temminckii): historical vicariance but little
present genetic structure in a regionally endangered Palearctic wader. Diversity and Distributions
18:704-716.
Seutin, G., B. White, and P. Boag. 1991. Preservation of avian blood and tissue samples for DNA
analyses. Canadian Journal of Zoology 69:82-90.
Wenink, P. W., and A. J. Baker. 1996. Mitochondrial DNA lineages in composite flocks of migratory and
wintering dunlins (Calidris alpina). The Auk 113:744-756.
Wenink, P. W., A. J. Baker, H. U. Rosner, and M. G. J. Tilanus. 1996. Global mitochondrial DNA
phylogeography of holarctic breeding dunlins (Calidris alpina). Evolution 50:318-330.
Wenink, P. W., A. J. Baker, and M. G. J. Tilanus. 1993. Hypervariable-control-region sequences reveal
global population structuring in a long-distance migrant shorebird, the dunlin (Calidris alpina).
Proceedings of the National Academy of Sciences of the United States of America 90:94-98.
Wenink, P. W., A. J. Baker, and M. G. J. Tilanus. 1994. Mitochondrial control-region sequences in two
shorebird species, the turnstone and the dunlin, and their utility in population genetic studies.
Molecular Biology and Evolution 11:22-31.
Wennerberg, L. 2001. Breeding origin and migration pattern of dunlin (Calidris alpina) revealed by
mitochondrial DNA analysis. Molecular Ecology 10:1111-1120.
Wennerberg, L., N. M. A. Holmgren, P. E. Jönsson, and T. von Schantz. 1999. Genetic and
morphological variation in dunlin (Calidris alpina) breeding in the Palearctic tundra. Ibis 141:391-
398.
Wennerberg, L., M. Klaassen, and A. Lindström. 2002. Geographical variation and population structure in
the white-rumped sandpiper (Calidris fuscicollis) as shown by morphology, mitochondrial DNA
and carbon isotope ratios. Oecologia 131:380-390.
Wilson, W. H. 1994. Western sandpiper (Calidris mauri), The Birds of North America Online. (A. Poole,
Ed.). Cornell Lab of Ornithology, Ithaca. Retrieved from the Birds of North America Online.
Available at http://bna.birds.cornell.edu.proxy.lib.sfu.ca/bna/species/090.
Port Metro Vancouver Hemmera RBT2 – WESA Genetics - 23 - December 2014
8.0 STATEMENT OF LIMITATIONS
This report was prepared by Hemmera Envirochem Inc. (“Hemmera”), based on fieldwork conducted by
Hemmera, for the sole benefit and exclusive use of Port Metro Vancouver. The material in it reflects
Hemmera’s best judgment in light of the information available to it at the time of preparing this Report.
Any use that a third party makes of this Report, or any reliance on or decision made based on it, is the
responsibility of such third parties. Hemmera accepts no responsibility for damages, if any, suffered by
any third party as a result of decisions made or actions taken based on this Report.
Hemmera has performed the work as described above and made the findings and conclusions set out in
this Report in a manner consistent with the level of care and skill normally exercised by members of the
environmental science profession practicing under similar conditions at the time the work was performed.
This Report represents a reasonable review of the information available to Hemmera within the
established Scope, work schedule and budgetary constraints. The conclusions and recommendations
contained in this Report are based upon applicable legislation existing at the time the Report was drafted.
Any changes in the legislation may alter the conclusions and/or recommendations contained in the
Report. Regulatory implications discussed in this Report were based on the applicable legislation existing
at the time this Report was written.
In preparing this Report, Hemmera has relied in good faith on information provided by others as noted in
this Report, and has assumed that the information provided by those individuals is both factual and
accurate. Hemmera accepts no responsibility for any deficiency, misstatement or inaccuracy in this
Report resulting from the information provided by those individuals.
APPENDIX A
Captured Western Sandpiper Morphometrics
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 1 - December 2014
Appendix A Morphometrics of Western Sandpipers Captured at Roberts Bank, Sturgeon Bank, and Boundary Bay, Delta, B.C., Spring 2012
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Boundary Bay 19-Apr 05:30 2601-03701 ASY C + F U 3 100.0 4 24.3 26.3 25.0 -
Boundary Bay 22-Apr 07:50 2601-03702 ASY C + F M 4 98.0 4 23.1 24.8 26.0 Yes
Boundary Bay 22-Apr 07:50 2601-03703 SY C + F M 2 100.0 2 21.8 26.1 26.0 -
Boundary Bay 22-Apr 07:50 2601-03704 SY C + F F 3 103.0 2 26.6 27.1 25.5 Yes
Boundary Bay 22-Apr 07:50 2601-03705 ASY C + F M 2 96.0 4 22.2 25.3 25.0 -
Boundary Bay 22-Apr 07:50 2601-03706 ASY C + F M 2 98.5 4 23.2 24.6 23.0 -
Boundary Bay 22-Apr 07:50 2601-03707 ASY? C F 2 103.0 3 27.2 31.2 26.0 Yes
Boundary Bay 22-Apr 07:50 2601-03708 SY C + F M 2 92.0 2 20.4 25.3 25.0 Yes
Boundary Bay 22-Apr 08:35 2601-03709 ASY C + F M 3 101.0 4 23.5 25.5 25.0 -
Boundary Bay 22-Apr 08:35 2601-03710 ASY C + F M 2 95.0 4 21.4 25.1 23.0 -
Boundary Bay 22-Apr 08:35 2601-03711 SY C + F M 2 97.5 3 22.2 25.5 24.0 Yes
Boundary Bay 22-Apr 08:35 2601-03712 AHY C + F M 1 94.0 3 21.5 25.2 24.0 -
Boundary Bay 22-Apr 10:05 2601-03713 ASY C + F M 3 99.0 4 23.6 25.7 31.0 Yes
Boundary Bay 24-Apr 05:45 2601-03714 ASY C + F M 3 98.0 5 21.8 24.4 24.5 Yes
Boundary Bay 24-Apr 06:10 2601-03715 ASY C + F M 3 94.0 4 22.9 24.9 25.5 -
Boundary Bay 24-Apr 06:10 2601-03716 ASY C + F F 2 102.0 5 25.8 26.8 26.0 Yes
Boundary Bay 24-Apr 06:10 2601-03717 ASY C + F M 3 99.0 4 22.6 26.3 24.5 -
Boundary Bay 24-Apr 06:10 2601-03718 SY C + F M 2 97.0 3 22.4 25.3 22.0 -
Boundary Bay 24-Apr 06:10 2601-03719 ASY C + F M 2 98.0 4 22.9 25.2 25.0 -
Boundary Bay 24-Apr 07:15 2601-03720 SY C + F M 4 97.5 3 23.2 25.7 28.5 -
Boundary Bay 24-Apr 07:15 2601-03721 ASY C + F F 4 101.0 4 27.2 26.3 29.0 Yes
Boundary Bay 24-Apr 07:15 2601-03722 SY C + F M 2 95.0 2 22.5 26.4 27.0 Yes
Boundary Bay 24-Apr 07:15 2601-03723 SY C + F F 3 99.0 3 26.3 26.2 26.0 Yes
Boundary Bay 24-Apr 07:15 2601-03724 ASY C + F M 3 96.0 4 21.2 23.8 24.0 Yes
Boundary Bay 24-Apr 07:15 2601-03725 SY C + F M 1 95.0 2 21.0 24.9 23.0 -
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 2 - December 2014
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Boundary Bay 24-Apr 07:15 2601-03726 ASY C + F F 3 98.0 4 25.8 26.0 28.0 Yes
Boundary Bay 24-Apr 07:15 2601-03727 SY? C + F M 5 100.0 2 21.6 25.1 27.0 Yes
Boundary Bay 24-Apr 07:15 2601-03728 ASY C M 3 98.5 3 22.8 25.4 25.0 -
Boundary Bay 24-Apr 07:15 2601-03729 ASY C + F F 2 100.0 4 26.2 27.6 26.0 Yes
Boundary Bay 24-Apr 07:15 2601-03730 ASY C F 3 100.0 3 27.8 26.4 27.0 -
Boundary Bay 24-Apr 06:10 Not Banded - - - - - - - - -
Boundary Bay 4-May 04:30 1401-71136 ASY C + F F 4 100.0 4 26.6 26.9 28.5 -
Boundary Bay 4-May 04:30 1401-71137 ASY C + F M 4 95.0 4 22.2 25.5 25.0 -
Boundary Bay 4-May 04:30 1401-71138 SY C + F M 4 94.0 3 23.8 25.2 29.0 -
Boundary Bay 4-May 04:30 1401-71139 SY C + F M 3 95.0 3 21.4 23.1 25.0 -
Boundary Bay 4-May 04:30 1401-71140 ASY C + F M 4 98.0 3 24.0 26.2 26.0 Yes
Boundary Bay 4-May 04:30 1401-71141 SY C + F M 4 95.0 3 23.7 25.2 24.0 Yes
Boundary Bay 4-May 04:30 1401-71142 ASY C + F F 4 99.0 4 25.1 25.1 29.0 Yes
Boundary Bay 4-May 05:30 2601-03751 SY C + F F 1 100.0 2 26.1 27.1 23.0 Yes
Boundary Bay 5-May 05:00 2601-03752 SY C + F M 0 96.0 2 23.3 25.7 21.0 Yes
Boundary Bay 5-May 05:00 2601-03753 ASY C + F M 4 97.0 4 22.5 25.2 24.0 -
Boundary Bay 5-May 05:00 2601-03754 ASY C + F M 2 96.0 4 23.8 25.4 23.0 Yes
Boundary Bay 5-May 05:00 2601-03755 ASY C + F M 3 99.0 4 22.4 25.2 23.0 -
Boundary Bay 5-May 05:00 2601-03756 ASY C + F M 1 96.0 3 21.6 24.1 21.0 -
Boundary Bay 5-May 05:00 2601-03757 ASY C + F F 0 103.0 4 27.2 24.9 25.0 -
Boundary Bay 5-May 05:00 2601-03758 ASY C + F F 3 102.0 4 28.2 27.0 27.0 Yes
Boundary Bay 5-May 05:00 2601-03759 ASY C + F F 3 102.0 4 27.1 27.4 25.0 -
Boundary Bay 5-May 05:00 2601-03760 ASY C + F F 3 101.0 4 27.1 26.4 27.0 -
Boundary Bay 5-May 05:00 2601-03761 ASY C + F F 4 102.0 4 27.0 24.5 29.0 -
Boundary Bay 6-May 04:00-6:00 1401-71143 ASY C F 3 105.0 3 28.3 27.3 26.5 -
Boundary Bay 6-May 04:00-6:00 1401-71144 ASY C + F M 4 93.0 4 22.2 23.7 24.0 Yes
Boundary Bay 6-May 04:00-6:00 1401-71145 ASY C F 3 103.0 3 26.4 26.3 25.0 -
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 3 - December 2014
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Boundary Bay 6-May 04:00-6:00 1401-71146 SY C + F M 2 101.0 2 24.1 26.1 24.0 Yes
Boundary Bay 6-May 04:00-6:00 1401-71147 SY C + F F 2 96.0 3 25.4 28.1 24.0 Yes
Boundary Bay 6-May 04:00-6:00 1401-71148 SY C + F M 4 97.0 1 23.1 25.8 25.5 -
Boundary Bay 6-May 04:00-6:00 1401-71149 ASY C + F F 3 103.0 4 29.5 26.8 26.5 Yes
Boundary Bay 6-May 04:00-6:00 1401-71150 ASY C + F F 4 103.0 3 26.6 26.8 28.0 -
Boundary Bay 6-May 04:00-6:00 2601-03762 ASY C + F F 2 102.0 4 26.2 26.5 23.0 -
Boundary Bay 6-May 04:00-6:00 2601-03763 ASY C + F F 4 100.0 4 27.0 25.9 28.0 -
Boundary Bay 6-May 04:00-6:00 2601-03764 ASY? C + F F 2 103.0 3 26.5 27.1 25.5 -
Boundary Bay 6-May 04:00-6:00 2601-03765 ASY C + F M 2 99.0 4 22.1 24.8 23.0 -
Boundary Bay 6-May 04:00-6:00 2601-03766 ASY C + F M 4 95.0 4 23.4 25.4 23.0 -
Boundary Bay 6-May 04:00-6:00 2601-03767 SY C + F M 2 92.0 3 22.2 24.5 21.5 -
Boundary Bay 6-May 04:00-6:00 2601-03768 SY C + F F 3 96.0 3 27.3 26.8 27.0 Yes
Boundary Bay 6-May 04:00-6:00 2601-03769 ASY C + F F 4 95.0 3 27.5 26.7 29.0 -
Boundary Bay 6-May 04:00-6:00 2601-03770 SY C + F M 3 95.0 2 24.0 25.5 22.0 -
Boundary Bay 6-May 04:00-6:00 2601-03771 ASY C + F M 3 100.0 4 22.9 25.0 23.0 -
Boundary Bay 6-May 04:00-6:00 2601-03772 SY C + F F 3 99.0 3 25.6 26.0 23.0 Yes
Boundary Bay 6-May 04:00-6:00 2601-03773 SY C + F M 3 92.0 2 22.8 24.9 23.0 -
Boundary Bay 6-May 04:00-6:00 2601-03774 ASY C + F F 3 103.0 4 25.4 25.9 26.0 -
Boundary Bay 6-May 04:00-6:00 2601-03775 SY C + F F 3 101.0 2 26.3 26.6 25.0 -
Boundary Bay 6-May 04:00-6:00 2601-03776 ASY C + F M 2 100.0 4 22.0 24.7 22.0 -
Boundary Bay 6-May 04:00-6:00 2601-03777 SY C + F F 1 99.0 3 26.5 26.1 23.0 -
Boundary Bay 6-May 04:00-6:00 2601-03778 ASY C + F M 2 98.0 3 23.3 24.9 23.0 -
Boundary Bay 6-May 04:00-6:00 2601-03779 SY C + F M 2 99.0 2 22.1 24.7 22.0 -
Boundary Bay 6-May 04:00-6:00 2601-03780 SY C + F M 2 98.0 2 21.3 24.9 22.0 -
Boundary Bay 6-May 04:00-6:00 2601-03781 ASY C + F F 2 103.0 4 27.2 28.1 27.0 -
Boundary Bay 6-May 04:00-6:00 2601-03782 SY C + F M 1 99.0 3 23.9 24.4 22.0 -
Boundary Bay 6-May 04:00-6:00 2601-03783 ASY C + F M 3 96.0 4 22.7 24.7 22.0 Yes
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 4 - December 2014
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Boundary Bay 6-May 04:00-6:00 2601-03784 SY C + F M 4 93.0 1 22.2 25.3 24.5 -
Boundary Bay 6-May 04:00-6:00 2601-03785 SY C + F M 4 90.0 3 20.3 23.5 24.0 -
Boundary Bay 6-May 04:00-6:00 2601-03786 ASY C + F F 4 102.0 3 27.2 26.2 25.0 -
Boundary Bay 6-May 04:00-6:00 2601-03787 SY C + F M 4 93.0 3 23.1 24.4 25.0 -
Boundary Bay 6-May 04:00-6:00 2601-03788 SY C + F F 4 95.0 2 25.0 25.4 26.0 -
Boundary Bay 6-May 04:00-6:00 2601-03789 ASY C + F F 4 96.0 4 25.9 25.8 27.0 Yes
Boundary Bay 6-May 04:00-6:00 2601-03790 ASY C M 2 102.0 3 23.6 26.7 27.0 -
Boundary Bay 6-May 04:00-6:00 2601-03791 ASY C + F M 2 95.0 3 21.7 24.1 22.0 -
Boundary Bay 6-May 04:00-6:00 2601-03792 SY C + F M 2 101.0 3 22.5 28.9 27.0 -
Boundary Bay 6-May 04:00-6:00 2601-03793 SY C + F M 2 95.0 2 22.9 24.8 21.0 -
Boundary Bay 6-May 04:00-6:00 2601-03794 SY C + F M 4 95.0 3 21.5 25.0 22.0 -
Boundary Bay 6-May 04:00-6:00 2601-03795 AHY C + F F 4 102.0 3 27.6 27.4 28.0 -
Boundary Bay 6-May 04:00-6:00 Not Banded ASY? C + F F 3 - 3 27.2 - - -
Boundary Bay 6-May 04:00-6:00 Not Banded - - - - - - - - - -
Boundary Bay 6-May 04:00-6:00 Not Banded SY C + F M - - - - - - -
Boundary Bay 6-May 04:00-6:00 Not Banded - - - - - - - - - -
Roberts Bank 24-Apr 18:30 2601-03801 ASY C + F M 5 98.0 3 22.2 25.2 30.0 Yes
Roberts Bank 24-Apr 18:30 2601-03802 ASY C + F M 4 103.0 4 23.7 25.0 30.5 Yes
Roberts Bank 24-Apr 18:30 2601-03803 ASY C F 4 100.0 - 27.2 26.3 29.0 Yes
Roberts Bank 24-Apr 18:30 2601-03804 ASY C + F M 4 97.0 4 22.0 24.2 23.0 Yes
Roberts Bank 24-Apr 18:30 2601-03805 ASY C + F M 3 97.0 3 22.5 25.9 27.0 Yes
Roberts Bank 24-Apr 18:30 2601-03806 ASY C + F M 3 101.0 4 22.2 25.4 26.0 -
Roberts Bank 24-Apr 18:30 2601-03807 ASY C + F M 4 96.0 4 22.2 26.1 28.0 Yes
Roberts Bank 24-Apr 18:30 2601-03808 SY C + F M 3 100.0 2 22.8 26.2 27.0 Yes
Roberts Bank 24-Apr 18:30 2601-03809 ASY C + F M 3 97.0 4 21.9 24.9 26.0 Yes
Roberts Bank 24-Apr 18:30 2601-03810 ASY C + F M 3 96.0 4 23.9 25.9 29.5 Yes
Roberts Bank 24-Apr 18:30 2601-03811 SY C M 4 99.0 4 21.4 23.5 25.5 Yes
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 5 - December 2014
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Roberts Bank 24-Apr 18:30 2601-03812 ASY C M 3 101.0 - 23.2 21.7 25.5 Yes
Roberts Bank 24-Apr 18:30 2601-03813 ASY C + F M 3 99.0 4 21.8 25.6 27.5 Yes
Roberts Bank 26-Apr 10:00 2601-03814 ASY C + F F 2 98.0 5 26.5 24.1 31.0 Yes
Roberts Bank 26-Apr 10:00 2601-03815 ASY C + F U 3 98.0 5 24.6 23.7 27.0 Yes
Roberts Bank 26-Apr 11:00 2601-03816 ASY C + F M 3 96.0 4 22.5 22.3 25.0 Yes
Roberts Bank 2-May 15:20 2601-03817 SY C + F M 1 91.0 2 21.9 25.4 23.0 Yes
Roberts Bank 2-May 15:20 2601-03818 SY C + F U 3 95.5 3 24.6 26.4 26.0 Yes
Roberts Bank 2-May 15:20 2601-03819 ASY C + F M 1 98.0 4 23.5 23.6 23.0 -
Roberts Bank 2-May 15:20 2601-03820 SY C + F F 3 98.0 4 26.0 26.3 32.0 -
Roberts Bank 2-May 15:20 2601-03821 ASY C + F M 2 91.0 4 22.7 26.6 26.0 Yes
Roberts Bank 2-May 15:20 2601-03822 SY C + F M 0 96.0 3 22.7 25.6 21.0 Yes
Roberts Bank 2-May 15:20 2601-03823 ASY C + F M 1 97.0 3 23.6 22.7 24.0 Yes
Roberts Bank 2-May 15:20 2601-03824 ASY C + F F 2 100.0 4 25.4 26.8 29.0 Yes
Roberts Bank 2-May 15:20 2601-03825 ASY C + F M 1 98.0 4 23.0 22.9 28.0 Yes
Roberts Bank 2-May 15:20 2601-03826 ASY C + F F 2 100.0 4 25.7 26.0 30.0 Yes
Roberts Bank 2-May 15:20 2601-03827 SY C + F M 2 91.0 3 25.0 23.7 29.5 Yes
Roberts Bank 2-May 15:20 2601-03828 ASY C + F F 1 97.0 4 26.9 26.9 28.0 Yes
Roberts Bank 2-May 15:20 2601-03829 SY C + F M 0 92.0 4 21.3 21.8 24.0 Yes
Roberts Bank 2-May 15:20 2601-03830 ASY C + F M 3 93.0 3 22.0 22.1 28.0 -
Roberts Bank 2-May 15:20 2601-03831 SY C + F F 1 96.0 3 25.8 27.0 26.0 Yes
Roberts Bank 2-May 15:20 2601-03832 SY C + F F 1 99.0 2 34.6 24.4 25.0 Yes
Roberts Bank 6-May 07:15 2601-03833 ASY C + F F 2 100.0 3 28.8 27.3 28.0 Yes
Roberts Bank 6-May 07:40 2601-03834 ASY C + F F 1 98.0 4 25.2 26.4 25.5 Yes
Roberts Bank 6-May 17:30 2601-03835 ASY C + F M 0 99.0 4 22.8 22.2 26.0 -
Roberts Bank 6-May 17:30 2601-03836 ASY C + F M 2 95.0 4 25.6 22.6 27.0 -
Sturgeon Bank 26-Apr 05:00 1401-66045 ASY C + F F 3 103.0 4 26.7 27.4 28.5 Yes
Sturgeon Bank 26-Apr 05:00 1401-79001 ASY C + F U 2 92.0 4 24.3 27.3 21.0 -
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 6 - December 2014
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Sturgeon Bank 26-Apr 05:00 1401-79002 ASY C + F M 4 96.0 - 22.9 24.7 28.5 Yes
Sturgeon Bank 26-Apr 05:00 1401-79003 ASY C + F F 4 99.0 - 27.5 27.3 27.0 -
Sturgeon Bank 26-Apr 05:00 1401-79004 ASY C + F F 2 102.0 - 25.3 26.0 26.0 -
Sturgeon Bank 26-Apr 05:00 1401-79005 ASY C + F F 3 101.0 4 26.0 26.6 30.0 -
Sturgeon Bank 26-Apr 05:00 1401-79006 ASY C + F F 4 103.0 4 28.4 26.9 30.0 -
Sturgeon Bank 26-Apr 05:00 1401-79007 ASY C + F M 4 93.0 3 22.5 24.1 25.0 Yes
Sturgeon Bank 26-Apr 05:00 1401-79008 ASY C + F F 3 103.0 3 26.1 27.0 28.0 -
Sturgeon Bank 26-Apr 05:00 1401-79009 SY C M 3 96.0 3 21.4 24.3 23.0 -
Sturgeon Bank 26-Apr 05:00 1401-79019 ASY C + F F 3 100.0 - 27.3 26.2 29.5 -
Sturgeon Bank 26-Apr 05:30 2601-03731 ASY C + F M 4 100.0 4 22.6 26.1 27.0 Yes
Sturgeon Bank 26-Apr 05:30 2601-03732 ASY C + F F 3 104.0 4 27.7 26.4 28.5 -
Sturgeon Bank 26-Apr 05:30 2601-03733 SY C + F M 2 96.0 3 21.7 25.8 24.0 -
Sturgeon Bank 26-Apr 05:30 2601-03734 ASY C F 3 108.0 3 27.5 27.6 28.5 -
Sturgeon Bank 26-Apr 05:30 2601-03735 ASY? C + F M 3 99.0 3 21.6 25.2 25.5 -
Sturgeon Bank 26-Apr 05:00 2601-03736 ASY C + F F 0 101.0 3 26.5 26.6 24.0 -
Sturgeon Bank 26-Apr 05:30 2601-03737 ASY C + F F 4 95.0 4 24.8 25.7 25.0 -
Sturgeon Bank 26-Apr 05:30 2601-03738 ASY C + F M 3 96.0 4 20.9 24.1 22.0 -
Sturgeon Bank 26-Apr 05:30 2601-03739 ASY C + F M 2 101.0 4 21.0 25.7 26.5 -
Sturgeon Bank 26-Apr 05:30 2601-03740 ASY C + F M 4 100.0 4 22.6 25.4 26.0 -
Sturgeon Bank 27-Apr 05:45 1401-66046 ASY C + F F 4 105.0 4 24.9 25.1 27.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-66047 ASY C + F F 4 101.0 2 27.0 26.0 28.0 -
Sturgeon Bank 27-Apr 05:45 1401-66048 ASY C + F F 3 104.0 4 27.8 26.5 28.0 -
Sturgeon Bank 27-Apr 05:45 1401-66049 ASY C + F F 3 100.0 4 26.9 27.0 29.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-66050 ASY C + F F 4 103.0 4 26.3 25.9 26.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71102 ASY C + F F 3 99.0 3 27.4 27.1 26.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71103 ASY C + F F 3 102.0 4 25.0 26.0 27.0 -
Sturgeon Bank 27-Apr 05:45 1401-71104 ASY C + F M 3 96.0 4 22.3 24.7 23.0 -
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 7 - December 2014
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Sturgeon Bank 27-Apr 05:45 1401-71105 ASY C F 1 102.0 3 28.0 26.8 22.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71106 SY C + F M 3 93.0 2 22.7 24.3 22.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71107 SY C + F M 2 95.0 2 23.2 25.2 23.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71108 ASY C + F F 3 98.0 4 26.5 25.4 23.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71109 ASY C + F F 1 102.0 3 25.4 25.8 24.0 -
Sturgeon Bank 27-Apr 05:45 1401-71110 ASY C + F M 3 100.0 3 22.9 25.0 25.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71111 SY C + F M 3 95.0 2 24.0 25.3 28.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-71120 ASY C F 4 101.0 3 27.0 26.1 26.0 Yes
Sturgeon Bank 27-Apr 05:45 1401-79010 ASY C + F F 4 103.0 4 25.7 25.6 27.0 -
Sturgeon Bank 27-Apr 05:45 1401-79011 ASY C + F F 2 99.0 3 25.2 26.5 24.0 -
Sturgeon Bank 27-Apr 05:45 1401-79012 ASY C + F M 2 100.0 4 22.9 24.4 26.0 -
Sturgeon Bank 27-Apr 05:45 1401-79013 ASY C + F M 2 99.0 4 23.7 24.8 24.0 -
Sturgeon Bank 27-Apr 05:45 1401-79014 ASY C + F F 3 102.0 4 24.8 25.7 26.0 -
Sturgeon Bank 27-Apr 05:45 1401-79015 ASY C + F M 3 99.0 4 24.1 25.3 25.0 -
Sturgeon Bank 27-Apr 05:45 1401-79016 ASY C + F F 3 99.0 3 27.2 27.2 25.0 -
Sturgeon Bank 27-Apr 05:45 1401-79017 ASY C + F U 2 100.0 4 24.7 27.6 25.0 -
Sturgeon Bank 27-Apr 05:45 1401-79018 ASY C + F F 0 102.0 3 24.8 25.2 24.0 -
Sturgeon Bank 2-May 04:30 1401-71112 ASY C + F F 2 97.0 5 25.5 24.6 22.5 -
Sturgeon Bank 2-May 04:30 1401-71113 ASY C + F M 3 98.0 4 23.8 26.0 25.0 Yes
Sturgeon Bank 2-May 04:30 1401-71114 SY C + F M 4 93.0 3 22.9 25.8 24.0 -
Sturgeon Bank 2-May 04:30 1401-71115 ASY C + F F 4 103.0 3 28.0 26.2 25.5 Yes
Sturgeon Bank 2-May 04:30 1401-71116 ASY C + F M 3 99.0 4 22.8 26.2 24.5 Yes
Sturgeon Bank 2-May 04:30 1401-71117 ASY C + F F 4 102.0 4 27.0 25.8 24.5 -
Sturgeon Bank 2-May 04:30 1401-71118 ASY C + F M 3 100.0 4 21.9 24.6 20.5 Yes
Sturgeon Bank 2-May 04:30 1401-71119 SY C + F M 4 96.0 2 22.9 26.0 26.5 Yes
Sturgeon Bank 2-May 04:30 1401-71122 SY C + F M 3 97.0 3 22.1 25.6 21.5 -
Sturgeon Bank 2-May 04:30 1401-71123 ASY C + F F 3 101.0 3 26.1 26.6 25.5 Yes
Port Metro Vancouver APPENDIX A Hemmera RBT2 – WESA Genetics - 8 - December 2014
Site Date Capture
time Band# Age
1 How aged
2 Sex3
Fat4 Wing
(mm)
Flight feather wear
5
Culmen (mm)
Tarsus (mm)
Weight (g)
DNA Analysed
Sturgeon Bank 2-May 04:30 1401-71124 ASY C + F M 3 97.0 4 22.4 24.4 21.5 Yes
Sturgeon Bank 2-May 04:30 1401-71126 ASY C + F F 2 99.0 4 27.1 26.8 25.5 Yes
Sturgeon Bank 2-May 04:30 1401-71127 ASY C + F F 4 103.0 3 29.0 26.7 26.5 Yes
Sturgeon Bank 2-May 04:30 1401-71128 ASY C + F F 4 102.0 4 25.8 26.5 24.5 Yes
Sturgeon Bank 2-May 04:30 1401-71129 ASY C + F M 4 96.0 4 20.4 24.5 22.5 -
Sturgeon Bank 2-May 04:30 1401-71130 ASY C + F U 3 101.0 3 24.7 26.7 24.0 Yes
Sturgeon Bank 2-May 04:30 1401-71131 ASY C + F M 4 91.0 4 21.8 25.6 24.0 Yes
Sturgeon Bank 2-May 04:30 1401-71132 ASY C + F U 3 100.0 4 24.2 26.2 23.5 Yes
Sturgeon Bank 2-May 04:30 1401-71133 ASY C + F M 4 96.0 3 22.6 25.2 25.0 Yes
Sturgeon Bank 2-May 04:30 1401-71134 ASY C + F U 3 98.0 3 24.0 25.4 23.0 -
Sturgeon Bank 2-May 04:30 1401-71135 ASY C + F F 2 99.0 4 26.1 26.5 24.5 Yes
Notes: 1 Age: SY = second year (born previous year), ASY = after second year, AHY = after hatch year.
2 How Aged: C = coverts, F = feather wear.
3 Sex: M = male, F = female, U = indeterminable/unknown (culmen length: F > 24.7 mm, M < 24.2 mm).
4 Fat: 0 = none, 1 = trace covering of furculum, 2 = furculum trace to 50% full, 3 = furculum 50% to 100% full, 4 = furculum beginning to bulge, abdomen full, 5 = furculum obviously bulging, abdomen bulging, 6 = fat overflowing furculum, abdomen, in armpits, 7 = bird covered in fat.
5 Flight feather wear: 0 = heavily abraded, shafts broken, feather tips missing, 1 = all feathers tips abraded and worn; large pieces of feather tips missing, 2 = numerous nicks and wear, 3 = one of two shallow nicks in one or two feathers, 4 = minimal wear, 5 = fresh feathers, brand new, no wear.