R/V William Kennedy Leg 4 Cruise Report (Sep 1-15, 2019) Cruise … · 2020. 11. 13. · agreement...

R/V William Kennedy Leg 4 Cruise Report (Sep 1-15, 2019)

Cruise along the Kivalliq transportation corridor (Hudson Bay)

September, 2019

Winnipeg, MB

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List of Participants

University of Manitoba

Paloma Carvalho (Chief Scientist)

Glen Hostetler (Research Associate)

Durell Desmond (PhD student)

Cathrin Veenaas (Post-doc)

Keesha Peterson (Technician)

McGill University/ National Research Council Canada (NRC)

Tammy Cai (Post-doc)

Alexa Bakker (Post-doc)

University of Calgary

Alastair Smith (Post-doc)

Meng Ji (MSc student)

Université Laval

Camille Lavoie (MSc student)

Environment and Climate Change Canada (ECCC)

Jasmin Schuster (Researcher)

Community member

Barbara Katorka

Ship crew

David McIsaac (Captain)

Daniel McIsaac (First mate)

Tyson Arsenault (Bridge watch)

Matthew Rose (Bridge watch)

Billy Gaudet (Cook)

Anne-Louise Dauphinee (Bridge watch and Logistics coordinator)

James Hill-Stosky (SVOP small boat operator)

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Table of Contents

List of Participants .............................................................................................................. 2

Table of Contents ................................................................................................................ 3

List of Tables ...................................................................................................................... 5

List of Figures ..................................................................................................................... 6

1. Chief Scientist Report ................................................................................................. 7

1.1 Summary .............................................................................................................. 7

2. Baseline monitoring of microbial genomics along the Kivalliq transportation

corridor (MPRI) and Microbial genomics for oil spill preparedness in Canada’s Arctic

marine environment (GENICE) .......................................................................................... 9

2.1 Introduction and Objectives ................................................................................. 9

2.2 Operations Conducted and Methodology ........................................................... 10

2.2.1 Sediment Core Sampling ............................................................................ 10


corridor .............................................................................................................................. 13

3.1 Introduction ........................................................................................................ 13


4. Investigation of ice adhering bacteria-diatom symbiosis and their interactions with

spilled oils ......................................................................................................................... 16

4.1 Introduction ........................................................................................................ 16


5. Non-target screening of Arctic environmental samples for the detection of unknown

organic pollutants – Spatial and temporal distribution across multiple environmental

compartments .................................................................................................................... 17

5.1 Introduction and Objectives ............................................................................... 17


5.2.1 Sampling ..................................................................................................... 19

5.2.2 Sample analysis ........................................................................................... 22

5.2.3 Data analysis ............................................................................................... 22

5.3 References .......................................................................................................... 23

6. Monitoring of organic pollutants and microplastics in air, water and sediment ....... 23



6.2.1 Atmospheric samples for organic pollutants ............................................... 23

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6.2.2 Water Particulate Samples .......................................................................... 25

6.2.3 Water sampling at stations .......................................................................... 26

6.2.4 Sediment samples for organic pollutants and microplastics ....................... 29

7. Microbial Genomics for Oil Spill Preparedness in Canada’s Arctic Marine

Environment (GENICE) ................................................................................................... 31



7.2.1 Rosette water sample collection and processing ......................................... 32

7.2.2 Nutrient sample collection .......................................................................... 33

7.2.3 DOC and POC sample collection ............................................................... 33

7.2.4 Microbial biomass for DNA sequencing (vacuum pump method) ............. 34

7.2.5 Fixation for cell counting ............................................................................ 35

7.2.6 Zodiac sample collection and processing ................................................... 35

7.2.7 Box core sampling ...................................................................................... 38

7.2.8 Microcosm setup (water) ............................................................................ 40

7.2.9 Microcosm setup (water and sediment) ...................................................... 41

7.3 References .......................................................................................................... 42

8. Benthic biodiversity, biological productivity and biogeochemistry in western

Hudson’s Bay .................................................................................................................... 42



8.3 References .......................................................................................................... 49

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List of Tables

Table 1.1 Summary of sample collection during the R/V William Kennedy Leg 4 cruise.

............................................................................................................................................. 9 Table 2.1 Sediment push core retrieval site locations. ..................................................... 10 Table 2.2. Gravity sediment core retrieval site locations. ................................................ 12

Table 3.1 Samples collected from each station ................................................................ 16 Table 5.1 Samples collected during the R/V William Kennedy cruise 2019 (Leg 4). ..... 20 Table 5.2 Coordinates for biota, water and surface sediment sampling. ......................... 21 Table 5.3 Coordinates for sediment sampling using the Zodiac ...................................... 21 Table 6.1 Sampling information for atmospheric samples. ............................................. 24

Table 6.2 Sampling information for particulate samples ................................................. 26 Table 6.3 Sampling information for high volume water samples (HVW) and low volume

water samples for microplastics (MPW), perfluorinated compounds (PFC) and

organophosphate flame retardants (OPE) ......................................................................... 28 Table 6.4 Sampling information for sediment samples.................................................... 30 Table 7.1 Total samples collected, both from ship-based sampling and zodiac sampling.

........................................................................................................................................... 31 Table 7.2 Water samples collected from rosette. ............................................................. 34 Table 7.3 Locations of zodiac sampling sites. ................................................................. 37

Table 7.4 Box core sample locations. .............................................................................. 39 Table 7.5 List of incubations prepared. ........................................................................... 40

Table 7.6 Microcosm experiment design. ........................................................................ 41 Table 7.7 Surface water collection for microcosm. ......................................................... 42 Table 7.8 Surface sediment collection for microcosm. .................................................... 42

Table 8.1 Samples collected from the box core during Leg 4 of the 2019 R/V William

Kennedy Research Cruise ................................................................................................. 45

Table 8.2 Organisms collected from the benthic trawl during Leg 4 of the 2019 R/V

William Kennedy Research Cruise. .................................................................................. 46

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List of Figures

Figure 1.1 Cruise track and stations sampled during the R/V William Kennedy Leg 4

cruise in 2019. ..................................................................................................................... 8

Figure 1.2 Zodiac stations sampled during the R/V William Kennedy Leg 4 cruise, 2019.

............................................................................................................................................. 8

Figure 2.1 Sediment sampling using the box core (A); and the core tube inserted into the

box core (B), being retrieved (C) and measured (D) ........................................................ 11

Figure 2.2 Core being sectioned (A) using a measure ring (B) and stored in Whirl-Packs

bags (C) ............................................................................................................................. 11

Figure 2.3. Gravity core being deployed (A); and core sample being retrieved (B) and

measured (C) ..................................................................................................................... 12

Figure 3.1 Collection of water from Rosette .................................................................... 14

Figure 3.2 Real-time data from Seabird 19Plus SeaCAT profiler ................................... 15

Figure 3.3 Vacuum filtration setup .................................................................................. 15

Figure 5.1 Possible distribution of pollutants within the marine environment ................ 18

Figure 5.2 Sampling stations. S: Sediment, B: Biota, W: Water, Z: Zodiac sampling

station. ............................................................................................................................... 19

Figure 5.3 Large volume water sample collection ........................................................... 20

Figure 5.4 Surface sediment collection ............................................................................ 21

Figure 6.1 High-volume air sampler ................................................................................ 24

Figure 6.2 Map illustrating the range of the individual atmospheric samples. ................ 25

Figure 6.3 Particle sampler housing and glass fibre filter (top), removing filter after

filtration (bottom).............................................................................................................. 25

Figure 6.4 Map illustrating the range of the individual particle samples. ....................... 26

Figure 6.5 High volume water collection and extraction ................................................. 27

Figure 6.6 Map with sampling information for HVW (red) and low volume water

samples for MPW (white), PFC (green) and OPE (yellow). ............................................ 27

Figure 6.7. Low volume water sampling with a stainless steel bucket ............................ 29

Figure 6.8 Map with sampling information for surface sediment.................................... 30

Figure 7.1 Rosette water sample collection ..................................................................... 32

Figure 7.2 Collecting water from Niskin bottle into syringe ........................................... 33

Figure 7.3 Zodiac sampling setup .................................................................................... 35

Figure 7.4 Filtration through Sterivex filter ..................................................................... 36

Figure 7.5 Surface sediment collection on board zodiac ................................................. 38

Figure 7.6 Surface sediment collection via syringe ......................................................... 39

Figure 7.7 Example of the set-up for water-accommodated fraction preparation with

water from station RI3. The aspirator bottle is being gently stirred on a magnetic stir

plate. The bottles were normally covered with foil to minimise photo-degradation but the

foil was removed to allow the photo to be taken. ............................................................. 41

Figure 8.1 Benthic trawl deployment ............................................................................... 44

Figure 8.2 Biota sorting ................................................................................................... 44

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1. Chief Scientist Report

Paloma Calabria Carvalho1

1Centre for Earth Observation Science, University of Manitoba, Winnipeg MB, Canada

R3T 2N2

1.1 Summary

Leg 4 of the 2019 R/V William Kennedy expedition along the Kivalliq

transportation corridor in western Hudson Bay was very successful. Most of our sampling

stations proposed in earlier meetings were achieved, while only a few stations located in

proximity to sampled stations were skipped. The decision to skip stations was made in

agreement during our regular science meetings (daily) aboard the R/V William Kennedy.

The science crew arrived in Coral Harbour on Aug 31, 2019 and boarded the R/V

William Kennedy later in the afternoon. On Sep 1, a community member from Naujaat,

Barbara Katorka, joined the vessel as part of the GENICE/MPRI projects. On Sep 1, we all

had familiarization with the vessel and safety instruction conducted by the first mate,

Daniel McIsaac. On the same day, we started mobilization and set up the lab space. Due to

weather, our cruise was delayed and we left Coral Harbour in the evening of Sep 2.

However, we sampled at our first station in the afternoon of Sep 2 in proximity to Coral

Harbour (CH0). The weather was good for the rest of the cruise and we managed to

complete 25 stations, sampling for water, sediment and benthic organisms (Fig. 1.1; Table

1.1). In addition to our full stations, 7 zodiac stations (with 5 substations each; see section

7.2.6 for details) were sampled for water and sediment (Fig. 1.2). As scheduled, we ended

our cruise in Churchill on Sep 15, where we worked on demobilization and shipping

supplies and samples back to the various involved institutions for future analyses.

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Figure 1.1 Cruise track and stations sampled during the R/V William Kennedy Leg 4

cruise in 2019.

Figure 1.2 Zodiac stations sampled during the R/V William Kennedy Leg 4 cruise, 2019.

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Table 1.1 Summary of sample collection during the R/V William Kennedy Leg 4 cruise.

Station ID Lat (N) Lon (W) Depth (m) Date Water Sediment Benthic

CH0 64°06.744' 83°13.371' 14.5 2-Sep X X

CH1 63°12.243' 83°49.595' 112 3-Sep X X X

CH2 63°15.593' 88°20.177' 182 4-Sep X X X

CII1 63°23.405' 90°43.456' 54 4-Sep X

CII4 63°42.856' 92°00.655' 58 5-Sep X X X

CII5 63°59.021' 93°30.445' 29.4 6-Sep X X X

CII7 63°55.981' 93°36.749' 37.7 6-Sep X X

BL1 64°06.660' 94°24.076' 67 7-Sep X X X

BL2 64°12.259' 95°08.498' 33.2 8-Sep X

BL3 64°13.808' 95°28.338' 37.7 7-Sep X X X

BL5 64°18.344' 95°59.921' 24.9 8-Sep X X

BL4 64°18.005' 95°57.421' 29.9 8-Sep X

CII10 63°55.018' 93°58.394' 40.9 8-Sep X X

CI3 63°02.419' 90°29.675' 36.5 9-Sep X X X

CI1 63°20.078' 90°41.455' 11 10-Sep X X

CI2 63°19.066' 90°39.578' 33.7 10-Sep X X

RI3 62°48.788' 92°02.569' 22 11-Sep X X

RI6 62°47.978' 92°05.978' 27.5 11-Sep X X X

RI4 62°45.896' 92°01.604' 40.8 11-Sep X

RI1 62°42.001' 91°34.427' 38.6 11-Sep X

AV1 61°44.796' 92°31.575' 46.8 12-Sep X X X

AV2 61°06.526' 93°56.462' 12.6 12-Sep X X X

AV3 61°06.840' 94°01.542' 5.8 13-Sep X X X

AV4 60°47.393' 93°57.544' 45.5 13-Sep X X X

AV5 60°26.435' 94°23.293' 28.1 14-Sep X X X


corridor (MPRI) and Microbial genomics for oil spill preparedness in Canada’s

Arctic marine environment (GENICE)

Principal Investigator: Gary Stern1; Cruise participants: Paloma Calabria Carvalho1, Glen

Hostetler2, and Durell Sterling Desmond1


R3T 2N2

2Natural Resources Institute, University of Manitoba, Winnipeg MB, Canada R3T 2M6

2.1 Introduction and Objectives

The GENICE and MPRI projects seek to build a reliable baseline of sediment

contamination along the Kivalliq transportation corridor (western Hudson Bay) in

preparation for the high possibility of oil and/or fuel contaminants spilled into the Northern

Arctic due to present and continuing increases in ship traffic and potential future oil and

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gas exploration. Moreover, through the characterization of the spatial and temporal

distribution of contaminants in the sediment, this project aims to link the historic sediment

formation and resultant contaminant partitioning with the risk of exposure and vulnerability

to new potential sources of petroleum sources, helping to facilitate the development and

assess the potential of using bioremediation as a feasible spill mitigation response.

2.2 Operations Conducted and Methodology

2.2.1 Sediment Core Sampling

A box corer (625cm2 sampling area, 25cm x 25cm x 50cm box area) was used to

collect sediment cores at full stations where soft sediment was present (Table 2.1). A

multibeam sonar (Furuno FCV – 1200L) was used to assess the presence of soft sediment

with the absence of large rocks that could damage the box corer. The box corer was

deployed using the A-frame (5000kg capacity) and winches (340kg capacity) on the stern

of the ship (Fig. 2.1A). If the bottom of the box corer was sealed and the sediment inside

was not slumped, a core tube (9 cm inner diameter) was then pressed into the sediment by

hand and subsequently sealed at its surface and base (Fig. 2.1B). The sediment push core

was then taken to the deck on board the ship, measured for its length (e.g., Fig. 2.1C and

D), and sectioned into Whirl-Packs bags in intervals of 0.5 cm until 10 cm, 1 cm until 20

cm, 2 cm until 30 cm, and 5 cm intervals for the remainder of the core (Fig. 2.2).

Table 2.1 Sediment push core retrieval site locations.

Date Station Latitude Longitude Length (cm)

04-Sep CH2 63°15.173' -88°23.263' 8

04-Sep CH2 63°15.341' -88°22.508' 8

05-Sep CII4 63°44.237' -92°00.613' 13

05-Sep CII4 63°44.428' -92°00.566' 15

06-Sep CII5 63°59.276' -93°30.229' 17

06-Sep CII7 63°55.785' -93°36.827' 45

07-Sep BL1 64°06.589' -94°24.871' 19

07-Sep BL3 64°13.621' -95°28.184' 17

08-Sep BL5 64°18.345' -95°59.915' 37

08-Sep CII10 63°54.940' -93°57.351' 21

11-Sep RI6 62°47.705' -92°05.739' 17

11-Sep RI4 62°46.020' -92°01.606' 11

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Figure 2.1 Sediment sampling using the box core (A); and the core tube inserted into the

box core (B), being retrieved (C) and measured (D)

Figure 2.2 Core being sectioned (A) using a measure ring (B) and stored in Whirl-Packs

bags (C)

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Similarly, a gravity corer (6.66 cm inner diameter) was used to collect sediment

cores at full stations with exceptionally soft and deeper sediment (Table 2.2), based off the

previously retrieved box cores. The corer consisted of a metal pipe with two removable

linings of plastic tubing 75 cm in length each (Fig 2.3A). Heavy weights sat atop the pipe.

Cores were retrieved by hanging the gravity core from the A-frame ~1.2 m from the deck

and removing the removable plastic tubing (holding the sediment core) from the metal pipe

and subsequently sealing the open sides of the tube (Fig. 2.3B). The gravity core was then

measured for its length (Fig. 2.3C) and sectioned into Whirl-Packs bags in intervals of 1

cm until 20 cm, 2 cm until 30 cm, and 5 cm intervals for the remainder of the core.

Table 2.2. Gravity sediment core retrieval site locations.

Date Station Latitude Longitude Length (cm)

05-Sep CII4 63°44.237' -92°00.729' 37

06-Sep CII5 63°59.296' -93°30.280' 53

06-Sep CII7 63°55.955' -93°36.739' 87

06-Sep CII7 63°55.962' -93°37.020' 70

07-Sep BL3 64°13.699' -95°28.206' 90

11-Sep RI6 62°47.808' -92°05.685' 67

Figure 2.3. Gravity core being deployed (A); and core sample being retrieved (B) and

measured (C)

The Whirl-Packs bags for both the push and gravity cores were then placed into a

-20°C freezer and sent to the University of Manitoba for radioisotope dating and

contaminant analyses (i.e., total mercury, methylmercury, hydrocarbons, and heterocyclic

compounds). Based on these analyses, an assessment as to the background contamination

levels at these site locations will be achieved. Furthermore, spatial and temporal trends of

the contaminants within the vertical distribution of the cores will be completed and

thoroughly investigated. Lastly, the differences in sediment depth as well as the levels and

types of contaminants found at each site will be compared and rationalized based on known

exposure history, topography of each location, and microbial presence and distribution.

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corridor

Principal investigator: Gary Stern1 and Charles Greer2,3; Cruise participants: Alexa

Bakker2,3and Tammy Cai2,3


R3T 2N2 2McGill University, Montreal, QC H3A 0G4 3National Research Council Canada (NRC), Montreal, QC H4P 2R2

3.1 Introduction

Western Hudson Bay communities experience high volumes of ship traffic due to

vast mineral exploration projects in the region. In Chesterfield Inlet, it is estimated that the

average change in shipping activity between 1990-2000 and 2011-2015 increased by 2000

– 4120 km. In the event of an accidental spill, one of the challenges will be discriminating

between natural background and contaminating hydrocarbons due to the spill. The project

will build a database containing detailed hydrocarbon and non-hydrocarbon contaminant

profiles (e.g. diagnostic ratios or “fingerprints”) and chemical concentrations in sediments

(particulate and bottom), water column samples and benthic invertebrates. Genomics

profiling will be conducted in sediments and water.


Under the collaborative framework, our team focuses on the development the

microbial genomics baseline database in the surface water as spilled oil slicks/plumes tend

to concentrate in the surface water. Sampling locations are selected to represent differences

in a) salinity, b) ship/tanker activity level, c) nutrient input, d) distance from pollution

sources from accidents/fuel farm/sewage, and e) ecology/community values (i.e. fishing,

whales, and bird sanctuaries) along the Kivalliq transportation corridor. Another objective

of our team is to isolate/culture oil degrading bacteria from the surface water samples

selected from the potentially polluted sites. Enrichment microcosms are used to enhance

the population dominancy of these oil degrading bacteria to facilitate the

isolation/culturing. IChip techique, which utilize an array of mini diffusion chambers to

provide the bacteria with the nutritional requirements mimicking their natural habitats, will

be utilized for the isolation/culturing.

Water samples will be collected using a CTD-Rosette system, which is deployed

from the R.V. William Kennedy (Figure 3.1). The R.V. William Kennedy is equipped with

a multibeam echo-sounder (WMB3250 WASSP Multibeam Sonar Survey Model with a

hull mounted WMB-T160S/20 WASSP Transducer 160Khz) which will be used

simultaneously for bottom mapping. All samples will be processed while on board and

shipped back to McGill University for analysis. This information will be invaluable in the

development of oil spill mitigation strategies, in assessing the success of remediation

strategies and in source attribution (i.e. to help establish ultimate responsibility for the

spill).

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Sampling was collected at 21 different stations (Table 3.1). At each station, water

was collected in the 12 bottle Rosette sampler at approximately 1.6m below the surface.

Attached to the Rosette was a Seabird 19Plus SeaCAT profiler which collected information

such as depth, salinity, oxygen content, and temperature and displayed it in real time

(Figure 3.2). A two-liter sample of water was harvested from each of three Rosette bottles

and the sample from each bottle was vacuum filtered through a different 47mm 0.22 µm

PES membrane using a Pall filtration manifold with disposable cups (Fig. 3.3). Each

membrane was immediately stored at -80°C for future genomic analyses. Samples will be

subjected to DNA and RNA extraction. DNA extracts will be subjected to 16S rRNA gene

amplification, library preparation and sequencing (Illumina). Generated sequences (fastq

files) will be used for downstream data processing. Operational taxonomic units (OTUs)

and the corresponding abundance will be generated to calculate phylogenetic trees and

biodiversity indices of microbial communities.

Another two-liter sample was harvested from one of the same Rosette bottles and

filtered in the same way but then stored in a 50mL falcon tube with 20mL of seawater at

4°C for culturing. Two additional 50mL aliquots of seawater from the same Rosette bottle

were also stored in 50mL falcon tubes at 4°C for culturing. From another of the three initial

Rosette bottles, an additional 50mL aliquot was transferred to a 125mL amber glass serum

bottle, enriched with 0.5mL Bushnell Haas Broth and 1.25µL marine diesel, and stored at

4°C to serve as a microcosm for future study through isolation and culturing of microbes

able to use the diesel as a substrate.

Figure 3.1 Collection of water from Rosette

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Figure 3.2 Real-time data from Seabird 19Plus SeaCAT profiler

Figure 3.3 Vacuum filtration setup

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Table 3.1 Samples collected from each station

Station Membranes for

genomics

Membrane + aliquots

for culturing

Microcosm

CH0 X X X

CH1 X

CH2 X

CII1 X X X

CII4 X X X

CII5 X X X

CII7 X X X

BL1 X X X

BL3 X X X

BL5 X X X

BL4 X X X

CII10 X

CI3 X X X

CI1 X X X

RI3 X X X

RI6 X X X

RI1 X X X

AV1 X X X

AV2 X X X

AV3 X X X

AV4 X

4. Investigation of ice adhering bacteria-diatom symbiosis and their interactions

with spilled oils

Principal investigator: Charles Greer1,2; Cruise participants: Alexa Bakker1,2 and Tammy

Cai1,2

1McGill University, Montreal, QC H3A 0G4 2National Research Council Canada (NRC), Montreal, QC H4P 2R2

4.1 Introduction

Ice algae play a critical role in primary production and serve as part of the base of

the polar food web. An emerging group of ice-binding proteins produced by bacteria has

recently been discovered that enables bacteria to adhere to the surface of ice as well as

photosynthetic diatoms. In terms of ecological significance, this symbiotic process

involving bacteria, diatoms and ice would enhance solar energy conversion and nutrient

cycling under the ice. The presence of this symbiosis at the ice-seawater interface makes it

especially susceptible to the impact of potential oil spills in icy conditions which also tend

to concentrate at the ice-seawater interface.

So far, only Marinomonas primoryensis has been observed utilizing “ice adhesins”.

Although “ice adhesins” have been examined in vitro with pure cultures, they have yet to

be investigated in the context of indigenous microbial communities dwelling at the ice-

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seawater interface. Our team also aims to isolate and culture other potential ice adhering

bacteria from seawater samples and bottom ice core samples harvested at different

locations in the Canadian Oceans. We are especially interested in isolating and culturing

ice adhering bacteria that can also degrade petroleum substances. We use iChip technique

to improve the culturability of the obtained samples and culture novel isolates. Our team

also has been investigating how the presence of the oil affect the formation of this ice

adhering bacteria-diatom symbiosis.


Following extraction and DNA sequencing, which was described in the previous

section (Section 3.2), representative samples of major clusters will be selected for shotgun

metagenomic sequencing, based on the results of UPGMA (unweighted pair group method

with arithmetic mean) cluster analysis of the 16S rRNA gene dataset. Genes encoding two

types of ice adhesins will be studied: Type I with repeats-in-toxin (RTX)-like ice-binding

domains; and Type II containing Domain-Of-Unknown-Function (DUF) 3494 for ice

binding. To gain insights on the factors regulating this “strategic” ecological process, the

relative abundance of ice adhesion encoding genes was correlated with data on chlorophyll

a, bacterial abundance, particulate organic carbon, dissolved organic carbon, dissolved

nitrogen, macro-nutrients and salinity.

The same protocol for culturing and isolation in the previous section were used to

culture and enrich ice adhering and oil degrading bacteria. The isolates will be screened

using motility test. High mobility isolates will be examined under microscope to investigate

their interactions with ice crystals.

5. Non-target screening of Arctic environmental samples for the detection of

unknown organic pollutants – Spatial and temporal distribution across multiple

environmental compartments

Principal Investigator: Gary Stern1; Cruise participant: Cathrin Veenaas1


R3T 2N2


More than 100,000 different chemicals are being produced and used every day [1].

Some of these chemicals might reach the environment and have direct adverse effects, i.e.

are toxic, while others might enrich in biota (bioaccumulation) or the food chain

(biomagnification). More than 100 million tons of chemicals that are hazardous to the

environment are produced annually in Europe alone [2]. Especially for those chemicals

that can have negative impacts on the environment, it is important to assess their occurrence

in the environment.

Most studies and regular monitoring campaigns focus on a limited number of

compounds (targeted analysis) since untargeted methods are considered too time

consuming. However, to be able to not only detect selected chemicals in commerce but to

include the large numbers of chemicals and also their degradation products in the

environment, untargeted methods are needed. To achieve a comprehensive screening of

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“unknown” organic compounds in environmental samples, sophisticated analytical

techniques and advanced data analysis techniques must be used. Large amounts of

information can be generated using advanced analytical instruments that will,

consequently, be handled in a newly developed automated way involving computational

methods.

This project plans to identify organic pollutants in the Arctic environment to help

understand where there are shortcomings in the present understanding of chemicals’

environmental fate, and what differences exist between different environmental

compartments and different regions of the Canadian Arctic. Special emphasize will be put

on temporal and spatial trends of newly emerging (and so far not detected) compounds in

the Hudson Bay area. The results will support the understanding of chemicals reaching

different parts of the environment, and, ultimately, the food chain. Due to remoteness, low

biological diversity, and low ambient temperatures, Arctic areas are especially vulnerable

to contamination [3]. Moreover, although Arctic environments are often remote and not

close to industry, the Arctic is considered a sink for global pollutants [4].

Within the marine environment, compounds can move between compartments as

shown in Figure 4.1. The distribution of compounds between these compartments depends

on their physico-chemical properties. The aim of this study is the analysis of organic

pollutants within all three compartments of the marine environment: water, sediment and

biota. Overlaps and differences among the compartments will be identified and spatial and

temporal trends within the Canadian Arctic will be studied. During the R/V William

Kennedy sampling cruise, sediment cores for time-trend analysis were collected. Assessing

temporal trends of unknown compounds will show which compounds have been enriching

in the environment in the past without our knowledge. Furthermore, surface sediment at

various locations (small scale and large scale transects) was collected for spatial trend

analysis to show which parts of the Arctic are affected most by pollution. In addition, at 8

stations all three sample types, biota, water and surface sediment, were collected to assess

the distribution of contaminants among different environmental compartments.

Figure 5.1 Possible distribution of pollutants within the marine environment

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In target analysis a limited number of compounds that are known and expected in

environmental samples is studied. In non-target analysis, however, no pre-assumptions are

made and, hence, compounds that we do not expect, including for example degradation

products or compounds that are not listed in production registers, can also be found and

identified. These unexpected compounds are called “unknown” here.


5.2.1 Sampling

Large volume water samples, sediment samples (surface and core), biota samples

and were taken at, in total, 20 stations (Table 5.1 and Table 5.2, Figure 5.2). The respective

sampling methods are described in section 6.1.3.1, 2.2.1 and 8.2, respectively. In brief,

roughly 300L of water was collected into stainless steel cans using an immersible pump

(Fig. 4.3) and subsequently filtered through an XAD cartridge at a rate of 300 mL/min. The

XAD was stored in the fridge until analysis. Sediment samples were collected using a box

core for most stations (Fig. 4.4). At Zodiac stations (labeled Z, Table 5.3), sediment was

collected using a ponar grab sampler. Sediment cores were extracted from the box core

using a push core and subsequently sliced into 0.5 cm slices up to 10 cm, 1 cm slices up to

20 cm and 2 cm thereafter. Gravity core samples were handled in a similar manner.

However, due to the larger disturbance of the upper layers the first 10 cm were sliced into

1 cm slices. Finally, biota samples were collected for about 15 minutes using a beam trawl

to catch benthic organisms. Only biota samples that could be identified on board were kept

for chemical analysis. All sediment and biota samples were kept in the freezer until

analysis.

Figure 5.2 Sampling stations. S: Sediment, B: Biota, W: Water, Z: Zodiac sampling

station.

20

Figure 5.3 Large volume water sample collection

Table 5.1 Samples collected during the R/V William Kennedy cruise 2019 (Leg 4).

*Samples were collected from a Zodiac.

Station Water

volume (L)

No. of surface

sediment

sample jars

Biota (no. of

species for

chemical analysis)

Push core Gravity core

CH1 304 1 21

CH2 285 1 7 1

CII4 285 1 5 1 1

CII5 304 1 1

CII7 1 1

CII10 1

BL1 304 1 1

BL4 1

BL5 304

CI1 1

CI2 9

CI3 304 1 4

RI6 304 1 5

AV1 1 5

AV2 1 6

AV3 4

AV5 1 10

Z1* 4 transects

Z4* 5 transects

Z7* 5 transects

21

Table 5.2 Coordinates for biota, water and surface sediment sampling.

Station Date Water Surface sediment Biota

Latitude Longitude Latitude Longitude Latitude Longitude

CH1 Sep 3 63°12.551’ -83°48.302’ 63°12.508' -83°47.842' 63°12.450' -83°48.154'

CH2 Sep 4 63°15.587’ -88°20.017’ 63°15.408' -88°22.119' 63°15.465' -88°19.716'

CH2 Sep 4 63°15.333' -88°22.564'

CII4 Sep 5 63°44.428’ -92°00.566’ 63°44.429' -92°00.563' 63°44.010' -92°00.383'

CII5 Sep 6 63°59.021’ -93°30.445’ 63°59.277' -93°30.233' 63°59.592' -93°29.297'

CII7 Sep 6 63°55.781' -93°36.827' 63°55.286' -93°36.950'

CII10 Sep 8 63°54.940' -93°57.351'

BL1 Sep 7 64°06.660’ -94°24.076’ 64°06.570' -94°23.891' 64°06.993' -94°24.396'

BL4 Sep 8 64°17.765' -95°57.364'

BL5 Sep 8 64°18.344’ -95°59.921’

CI1 Sep 10 63°20.074' -90°41.453'

CI2 Sep 10 63°18.565' -90°39.866'

CI3 Sep 9 63°02.416’ -90°29.675’ 63°02.599' -90°33.520' 63°02.753' -90°33.824'

RI6 Sep 11 62°47.978’ -92°05.978’ 62°47.667' -92°05.851' 62°47.713' -92°06.957'

AV1 Sep 12 61°44.071’ -92°32.397’ 61°44.320' -92°32.083' 61°44.067' -92°32.362'

AV2 Sep 12 61°06.566' -93°56.563' 61°06.170' -93°55.259'

AV3 Sep 13 61°06.680' -93°58.548'

AV5 Sep 15 60°26.618' -94°23.875' 60°26.917' -94°24.729'

Table 5.3 Coordinates for sediment sampling using the Zodiac

Sub-station

Z1 Z4 Z7

Latitude Longitude Latitude Longitude Latitude Longitude

1 63°45.961 -92°02.003 64°16.061 -96°08.486 62°51.314 -92°05.888

2 63°45.762 -92°01.599 64°16.426 -96°07.611 62°51.266 -92°05.414

3 63°45.345 -92°00.967 64°16.624 -96°06.494 62°51.190 -92°04.929

4 63°44.834 -92°00.810 64°16.840 -96°05.402 62°51.058 -92°04.452

5 64°17.826 -96°01.923 62°50.081 -92°02.871

Figure 5.4 Surface sediment collection

22

5.2.2 Sample analysis

Extraction methods for the different sample matrices will be developed in a way to

ensure good extraction recoveries for a large number of compounds. These will include the

use of different types of solvents (polar as well as non-polar).

For a comprehensive screening of organic pollutants in environmental samples,

several different analytical techniques are being used and combined. While smaller and

less polar compounds can be analyzed by gas chromatography (GC), larger or more polar

compounds will have to be analyzed using liquid chromatography (LC). By performing

GC as well as LC analysis a comprehensive analysis of the samples is obtained, and all

organic compounds can be captured.

The samples will be analyzed by GC×GC-HRMS for the detection of smaller, less

polar compounds. In comparison to conventional GC, GC×GC provides an increased peak

capacity and hence improves the separation and identification of unknown compounds. In

addition, characteristic elution patterns (one and two-dimensional GC retention times) can

be used to identify compounds. The benefit of HRMS over low resolution MS is the

increased resolution which results in the possibility of accurate mass measurements of

fragment ions. In addition to GC×GC-HRMS, atmospheric pressure (AP) GC will be used.

Due to its softer ionization, molecular ions are more readily obtained using APGC and a

molecular formula can be more easily obtained, whereas the GC×GC-HRMS instrument

gives characteristic fragmentation spectra. The combination of both will allow a better

identification of unknown compounds.

More polar or larger analytes will be analyzed using state-of-the-art LC-ion

mobility-MS/MS. Tandem mass spectrometry (MS/MS) can be used in LC-MS analysis to

obtain characteristic fragmentation spectra as they are obtained in GC-MS. Since LC-MS

ionization usually occurs under atmospheric pressure it is a soft ionization that generally

does not cause any fragmentation. Hence, fragmentation spectra need to be generated by

using MS/MS, which includes a fragmentation step between the two mass analyzers. The

addition of ion mobility measurements has two advantages that will both be used. Firstly,

ion mobility provides an additional dimension of separation improving the overall peak

capacity, i.e. the number of compounds that can be detected. And secondly, ion mobility

measurements provide the analyst with a value corresponding to a compound’s size and

shape (the collision cross section; CCS) which gives an additional identification point for

unknown compounds.

5.2.3 Data analysis

Finally, the obtained data, i.e. identified compounds, will be compared among all

three matrices and the different points across the Arctic, and similarities and differences

will be discussed. A principal component analysis (PCA) could then be used to identify

groupings among all samples and, moreover, identify differences among the sample types

(water, biota and sediment from different points in the Canadian Arctic). Additional

statistical methods for the evaluation and characterization of the data will be researched if

necessary. Moreover, oceanographic and biogeochemistry data (metadata) that were

collected throughout the sampling campaign will be used to identify the origin of the

detected and identified compounds, using, for example, information about ocean currents.

The collected sediment cores will be dated and used to perform time trend analyses.

Furthermore, the surface sediment samples (including Zodiac stations) will be used to

23

perform spatial trend analysis. In addition, an analysis of biomagnification will be

performed by analyzing biota from different trophic levels and studying potential

enrichment of certain pollutants.

5.3 References

[1] A.B.A. Boxall, C.J. Sinclair, K. Fenner, D. Kolpin, S.J. Maund, When synthetic

chemicals degrade in the environment, Environ. Sci. Technol. 38(2004) 368A–375A.

[2] European Commission, Eurostat; online data code: env_chmhaz, (2018).

[3] R. Gunnarsdóttir, P.D. Jenssen, P. Erland Jensen, A. Villumsen, R. Kallenborn, A

review of wastewater handling in the Arctic with special reference to pharmaceuticals and

personal care products (PPCPs) and microbial pollution, Ecol. Eng. 50 (2013) 76–85.

[4] Arctic Monitoring and Assessment Programme (AMAP), Chemicals of Emerging

Arctic Concern - Summary for Policy-makers, (2017). https://oaarchive.arctic-

council.org/handle/11374/1947.

6. Monitoring of organic pollutants and microplastics in air, water and sediment

Principal Investigator: Lisa Jantunen1; Cruise participant: Jasmin Schuster2

1Air Quality Processes Research Section, Science and Technology Branch, Environment

and Climate Change Canada (ECCC), Egbert, ON L0L1N0

²Air Quality Research Division, Science and Technology Branch, Environment and

Climate Change Canada (ECCC), Toronto, ON M3H 5T4


The purpose of this study was to determine the occurrence, concentrations, and gas

exchange of select organic pollutants. Compound classes of interest included: pesticides

(current use and legacy), flame retardants (FRs), perfluorinated compounds (PFCs), and

polycyclic aromatic compounds (PACs), which include polycyclic aromatic hydrocarbons

(PAHs). The goal was to use the air and water samples collected to set baseline

environmental concentration levels for PACs and select FRs, as well as to continue to

monitor concentration trends for compounds previously studied (PFCs, pesticides, and

select FRs). Air and water samples were paired and the gas exchange will be calculated for

priority pollutants in order to determine whether the water is acting as a source or a sink

for these compounds.

Samples collected for organic pollutants and microplastics onboard the R/V William

Kennedy will be processed and analysed in the laboratory at Centre for Atmospheric

Research Experiments (CARE), Environment and Climate Change Canada (ECCC).


6.2.1 Atmospheric samples for organic pollutants

Atmospheric samples were collected while the ship was underway. The High-

volume air sampler was mounted above the bridge toward the bow of the ship (Fig. 6.1).

Incoming air was pulled through a sample head, which contained a 0.45 micron quartz fiber

filter followed by a XAD resin column to sample the particulate and gaseous phases

https://oaarchive.arctic-council.org/handle/11374/1947https://oaarchive.arctic-council.org/handle/11374/1947

24

respectively. The sampler ran continuously with occasional shutdowns during major

sampling stations. The sample head was changed after 24-48 hours. Six samples were

collected during the expedition. Details on the sampling locations are reported in Table 6.1

and Figure 6.2. Samples were stored at -20°C.

Figure 6.1 High-volume air sampler

Table 6.1 Sampling information for atmospheric samples.

Sample ID Start Latitude Longitude Stop Latitude Longitude

WK19Air1 02/09/2019 64.116 -83.140 04/09/2019 63.390 -90.724

WK19Air2 04/09/2019 63.396 -90.735 07/09/2019 64.119 -94.393

WK19Air3 07/09/2019 64.306 -95.533 08/09/2019 63.995 -94.311

WK19Air4 08/09/2019 63.995 -94.311 11/09/2019 62.760 -91.979

WK19Air5 11/09/2019 62.760 -91.979 12/09/2019 61.747 -92.526

WK19Air6 12/09/2019 61.747 -92.526 14/09/2019 60.077 -94.067

25

Figure 6.2 Map illustrating the range of the individual atmospheric samples.

6.2.2 Water Particulate Samples

Particle samples were collected while underway. Seawater (2m depth) pumped

from the engine room to the saltwater tap on the starboard deck was filtered overnight using

a glass fibre filter (Figure 6.3). The flow rate ranged from 1.8-2.3 L/min, resulting in

sample volumes of 970 – 1500 L. Five samples were collected during the expedition.

Details on the sampling locations are reported in Table 6.2 and Figure 6.4. Samples were

stored at -20°C.

Figure 6.3 Particle sampler housing and glass fibre filter (top), removing filter after

filtration (bottom)

26

Table 6.2 Sampling information for particulate samples

Sample ID Start Latitude Longitude Stop Latitude Longitude Volume [L]

WK19Part1 03/09/2019 63.046° -85.365° 04/09/2019 63.259° -88.347° 1430

WK19Part2 05/09/2019 63.836° -93.356° 06/09/2019 63.984° -88.347° 1495

WK19Part3 07/09/2019 64.306° -95.998° 08/09/2019 64.306° -95.999° 1287

WK19Part4 11/09/2019 62.539° -91.367° 12/09/2019 61.747° -92.526° 972

WK19Part5 13/09/2019 61.113° -93.985° 13/09/2019 61.113° -93.986° 990

Figure 6.4 Map illustrating the range of the individual particle samples.

6.2.3 Water sampling at stations

6.2.3.1 High volume surface water for organic pollutants and non-target analysis

High volume surface water samples (300 L) were collected in stainless steel

canisters by the use of a submersible pump deployed from the starboard deck (Figure

6.5A). These samples were extracted by pumping the water collected through a resin

column (Figure 6.5B); care was taken to limit the flow rate (~300mL/min) to ensure all

compounds of interest were captured. Nine samples were collected at selected Basic

Stations, as outlined in Table 6.3 and Figure 6.6. Samples were stored at 4°C.

27

Figure 6.5 High volume water collection and extraction

Figure 6.6 Map with sampling information for HVW (red) and low volume water samples

for MPW (white), PFC (green) and OPE (yellow).

28

Table 6.3 Sampling information for high volume water samples (HVW) and low volume

water samples for microplastics (MPW), perfluorinated compounds (PFC) and

organophosphate flame retardants (OPE)

Station Type Date Latitude Longitude

CH0

MPW 02/09/2019 64.088° -83.255°

OPE 02/09/2019 64.088° -83.255°

PFC 02/09/2019 64.088° -83.255°

CH1 HVW 03/09/2019 63.209° -83.805°

MPW 03/09/2019 63.209° -83.805°

CH2 HVW 04/09/2019 63.260° -88.334°

CII1 MPW 04/09/2019 63.390° -90.724°

CII2 OPE 04/09/2019 63.390° -90.724°

PFC 04/09/2019 63.390° -90.724°

CII4

HVW 05/09/2019 63.740° -92.010°

MPW 05/09/2019 63.798° -92.011°

PFC 05/09/2019 63.798° -92.011°

CII5 HVW 06/09/2019 63.984° -93.507°

CII7 MPW 06/09/2019 63.933° -93.612°

PFC 06/09/2019 63.933° -93.612°

BL1

HVW 07/09/2019 64.110° -94.401°

MPW 07/09/2019 64.113° -94.395°

PFC 07/09/2019 64.113° -94.400°

BL5

HVW 08/09/2019 64.306° -95.999°

MPW 08/09/2019 64.306° -95.999°

OPE 08/09/2019 64.306° -95.999°

PFC 08/09/2019 64.306° -95.999°

CI3

HVW 09/09/2019 63.040° -90.491°

MPW 09/09/2019 63.040° -90.495°

PFC 09/09/2019 63.040° -90.495°

CI1 MPW 10/09/2019 63.335° -90.691°

RI3 MPW 11/09/2019 62.813° -92.043°

PFC 11/09/2019 62.813° -92.043°

RI6 HVW 11/09/2019 62.800° -92.100°

AV1

HVW 12/09/2019 61.735° -92.540°

MPW 12/09/2019 61.747° -92.526°

OPE 12/09/2019 61.747° -92.526°

PFC 12/09/2019 61.747° -92.526°

AV2 MPW 12/09/2019 61.109° -93.941°

29

6.2.3.2 Low volume surface water for organic pollutants

Low volume surface water samples were collected by the use of a stainless steel

bucket deployed from the starboard deck (Fig. 6.7). Amber glass bottles (4L) were used to

collect samples to be analysed for FRs, and polyethylene bottles (1L) for PFCs. Nine and

four samples were collected at selected Basic Stations for PFC and OPE analysis

respectively, as outlined in Table 6.3 and Figure 6.6. Samples were stored at 4°C.

Figure 6.7. Low volume water sampling with a stainless steel bucket

6.2.3.3 Low volume surface water for microplastics

Low volume surface water samples (38L) were collected in stainless steel canisters

by the use of a stainless steel bucket deployed from the starboard deck (Fig. 6.7). The water

was filtered through a 10 µm pore size polycarbonate filter and stored in aluminum foil

awaiting microscope analysis. Twelve filter samples were collected at selected Basic

Stations, as outlined in Table 6.3 and Figure 6.6. Samples were stored at 4°C.

6.2.4 Sediment samples for organic pollutants and microplastics

Surface sediment samples were collected from the box core (625cm2 sampling area,

25cm x 25cm x 50cm box area) in 250 mL glass jars. Thirteen samples were collected at

basic stations and six samples were obtained from Zodiac expeditions, as outlined in Table

6.4 and Figure 6.8. Samples were stored at -20°C.

30

Table 6.4 Sampling information for sediment samples.

Station Date Latitude Longitude Depth (m)

CH1 03/09/2019 63.208° -83.797° 112.0

CH2 04/09/2019 63.257° -88.369° 182.0

CII4 05/09/2019 63.704° -92.010° 32.5

Z#1 05/09/2019 63.766° -92.033° 9.2

Z#1 05/09/2019 63.763° -92.027° 4.2

Z#1 05/09/2019 63.756° -92.027° 6.3

CII5 06/09/2019 63.988° -93.504° 32.2

CII7 06/09/2019 63.930° -93.614° 37.7

BL1 07/09/2019 64.110° -94.415° 43.9

BL3 07/09/2019 64.227° -95.470° 37.0

Z#4 08/09/2019 64.268° -96.141° 4.5

Z#4 08/09/2019 64.274° -96.141° 6.5

Z#4 08/09/2019 64.274° -96.108° 6.0

BL5 08/09/2019 64.306° -95.999° 25.1

CI3 09/09/2019 63.043° -90.559° 32.3

CI1 10/09/2019 63.335° -90.691° 11.7

RI3 11/09/2019 62.813° -92.043° 21.6

RI6 11/09/2019 62.794° -92.098° 30.5

AV2 12/09/2019 61.111° -93.943° 10.9

Figure 6.8 Map with sampling information for surface sediment

31

7. Microbial Genomics for Oil Spill Preparedness in Canada’s Arctic Marine

Environment (GENICE)

Principal Investigator: Casey Hubert1; Cruise participants: Alastair Smith1, Meng Ji1

1 Geomicrobiology Group, Department of Biology, University of Calgary, Calgary, AB

T2N 1N4


The GENICE project aims to investigate the potential for natural remediation of an

oil spill should it happen in Canadian Arctic and sub-Arctic waters. As part of this effort,

we sought to examine the biogeography of oil spill degradation potential across the coastal

waters of western Hudson Bay. In particular, we were interested in whether there were

differences in the capacities of the in situ microbial communities from the various sites

sampled during this leg to respond to an oil spill and, if so, whether these differences were

predictable based upon a suite of measured parameters.

To address these questions, we set up microcosms amended with bunker C fuel oil

using surface seawater from 16 sites and with a standardised protocol that will allow us to

compare rates of biodegradation between sites. Water samples from up to 4 depths at each

station were filtered and surface sediment collected for 16S amplicon sequencing, with

selected samples being used for metagenomic sequencing. This will give us an

understanding of both the microbial community composition and functional potential.

Samples for further analyses were collected to provide a richer context within which to

interpret our microbial community and biodegradation data. In short, these parameters

were:

Inorganic nutrients (ammonium, nitrate, nitrite, phosphate, silicate)

Dissolved and particulate organic carbon and nitrogen (DOC/DON and POC/PON,

respectively, henceforth referred to simply as DOC and POC)

Coloured dissolved organic matter (CDOM)

Microbial cell counts

An overview of the samples collected is presented in Table 7.1.

Table 7.1 Total samples collected, both from ship-based sampling and zodiac sampling.

DNA filters (vacuum pump) 180

DNA filters (peristaltic pump) 55

DNA (sediment) 63

Bulk surface sediment 52

Cell counts 279

DOC 215

POC 95

CDOM 57

Nutrients 215

32


7.2.1 Rosette water sample collection and processing

The ship was equipped with a CTD-rosette fitted with twelve 5L Niskin bottles

(Fig. 7.1). Sensors on the CTD allowed it to capture profiles of chlorophyll fluorescence,

photosynthesis-active radiation (PAR) and dissolved oxygen concentration, in addition to

determining water temperature and salinity. At each station, we selected up to 4 depths

from which to collect samples based primarily on the temperature and salinity profiles

generated on the rosette downcast, collecting 2 Niskin bottles (10L) from each sampled

depth. Surface and bottom water were collected at every station, with the exception of

station AV3, where only surface water was collected. Intermediate depths were selected to

lie above and below the pycno- or thermocline, where one was evident from the CTD data.

If this was not the case, sampled depths were approximately evenly spaced through the

water column. See Table 7.2 for a summary of rosette water samples collected.

Figure 7.1 Rosette water sample collection

Water for nutrient and DOC/POC analyses was drawn from the Niskin bottles first

and stored separately before draining the bottles from each depth into 10L HDPE carboys.

This water, approximately 7-8L per sampled depth, was used for collection of DNA

samples, cell counts and setting up microcosms. With the exception of the samples

collected for nutrient analyses, all of the water samples we collected were passed through

a 200 μm Sefar Nitex screen, held in a 47 mm polycarbonate filter holder, in order to

remove large particles and organisms. All tubing, filter holders and the Nitex screen were

acid washed in 10% HCl prior to the expedition and were rinsed using reverse osmosis

water between sampling stations.

33

7.2.2 Nutrient sample collection

Water was drawn directly from the spigot of the Niskin bottles into an acid-washed

60mL plastic syringe (Fig. 7.2). These syringes were washed three times with sample water

before attaching a 25 mm Swinnex filter holder (EMD Millipore) fitted with a 25 mm GF/F

filter (Whatman GE). The filter was rinsed with 10-15 mL of sample water before rinsing

a 15mL acid-washed polypropylene centrifuge tube (Sarstedt) three times with filtered

sample water and filling the tube to approximately the 12 mL mark. Samples were flash-

frozen at -80 °C before being transferred to a -20 °C freezer. We collected triplicate nutrient

samples for each depth.

Figure 7.2 Collecting water from Niskin bottle into syringe

7.2.3 DOC and POC sample collection

Water drawn from the Niskin bottles was used to fill acid-washed 500 mL

polycarbonate bottles and refrigerated until further processing, which occurred within 2 –

3 hours of sample collection. Acid-washed silicone tubing and a 25 mm in-line filter holder

were first rinsed with sample water pumped using a Geopump peristaltic pump (Geotech

Environmental) down to the 500 mL mark on the collection bottle – approximately 50 mL.

A pre-combusted GF/F filter (4 hours at 450 °C) was then placed into the filter holder and

the remaining 500 mL of sample water pumped through it. The filtered water was used to

rinse and fill acid-washed 40 mL glass EPA vials. Three of these vials were frozen at -20

°C for DOC analysis while a fourth was stored at 4 °C and will be used for CDOM

determination. The filter was wrapped in pre-combusted aluminium foil and stored at -20

°C for POC analysis.

34

Table 7.2 Water samples collected from rosette.

Station Date

Time

Latitude Longitude Station

depth (m)

Depths sampled

CH0 02/09/2019

15:33

64° 06.744 -83° 13.371 14.5 Surface, 10m

CH1 02/09/2019

09:49

63° 12.243 -83° 49.595 112 Surface, 10m, 50m,

100m

CH2 04/09/2019

06:25

63° 15.565 -88° 20.688 182 Surface, 20m, 50m,

170m

CII1 04/09/2019

19:18

63° 23.547 -90° 43.681 51.2 Surface, 20m, 40m

CII4 05/09/2019

07:43

63° 42.928 -92° 00.649 54 Surface, 20m, 35m,

45m

CII5 06/09/2019

07:23

63° 59.021 -93° 30.445 29.5 Surface, 10m, 20m

BL1 07/09/2019

08:25

64° 06.606 -94° 24.098 51.8 Surface, 15m,

37.5m, 57m

BL3 07/09/2019

14:43

64° 13.808 -95° 38.338 51.8 Surface, 10m, 20m,

30m

BL5 08/09/2019

08:30

64° 18.345 -95° 59.917 24.9 Surface, 10m, 14m

CII10 08/09/2019

21:24

63° 54.987 -93° 58.244 47 Surface, 15m, 35m

CI3 09/09/2019

13:53

63° 02.375 -90° 29.692 37.3 Surface, 10m, 20m,

27m

CI1 09/09/2019

07:23

63° 20.076 -90° 41.454 11.9 Surface, 5m

RI3 11/09/2019

07:20

62° 48.796 -92° 02.571 22.1 Surface, 10m, 20m

RI6 11/09/2019

10:11

62° 47.781 -92° 05.978 25.9 Surface, 10m, 20m

AV1 12/09/2019

06:30

61° 44.795 -92° 31.572 46.7 Surface, 10m, 25m,

35m

AV2 12/09/2019

18:42

61° 06.515 -93° 56.622 12.6 Surface, 5m

AV3 13/09/2019

08:59

61° 06.689 -93° 56.558 5.3 Surface

AV4 13/09/2019

15:49

60° 47.327 -93° 57.505 47.1 Surface, 15m, 37m

AV5 14/09/2019

08:33

60° 26.443 -94° 23.337 28.1 Surface, 10m, 20m

7.2.4 Microbial biomass for DNA sequencing (vacuum pump method)

Two litres of water from each depth was filtered in triplicate onto 0.2 μm PES filters

(Pall) using a Pall Sentino vacuum manifold and vacuum pump. In some cases, the filters

clogged before two litres had been passed through, in which case the exact volume filtered

35

was recorded. Filters were folded using sterile forceps, placed into whirlpack bags and

stored immediately at -80 °C. This was repeated to obtain triplicate filters for each depth.

7.2.5 Fixation for cell counting

Sample water was transferred into 2 mL cryovials and fixed by adding 2% v/v

glutaraldehyde. After mixing, the vials were incubated at room temperature for 15 minutes

before being stored at -80 °C.

7.2.6 Zodiac sample collection and processing

At several sampling stations adjacent to major rivers, a zodiac was used to conduct

a 5-point transect starting from the river mouth and heading back towards the ship (Fig.

7.3). This sampling strategy aimed to capture a gradient of salinities and of riverine

influence in terms of dissolved organic matter. At each site in the transect, samples were

collected for the same suite of parameters as at the main station but only surface water was

collected. A summary of all locations sampled from the zodiac is presented in Table 7.3.

Salinity and water temperature were measured using a YSI ProPlus Multiparameter probe

and surface water collected using a portable peristaltic pump powered using a 12V non-

spillable lead-acid battery. The pump tubing was threaded through a 1m length of copper

pipe so that water could be pumped from a defined depth under water.

Figure 7.3 Zodiac sampling setup

Samples for inorganic nutrient, DOC, POC and CDOM analyses were collected in

an analogous manner to those processed on the ship except that water was pumped directly

through a 25 mm filter holder fitted with a pre-combusted GF/F filter, which eliminated

the need for secondary sample containers (syringes or bottles). The volume of water passed

through the filter was measured by running the filtered water plus washings into a

graduated 2L bottle up to the 500 mL mark. Note that nutrient and DOC samples were not

36

collected in triplicate from zodiac sites. Triplicate unfiltered samples for cell counts were

also collected at each site in 2mL cryovials and processed as described above on return to

the ship.

7.2.6.1 Microbial biomass for DNA sequencing (peristaltic pump method)

Surface water from each site was pumped into a graduated 2L polycarbonate bottle

up to the 2L mark. Then, again using the peristaltic pump, water from the bottle was

pumped through a 0.2 μm Sterivex filter (EMD Millipore) either until the full 2L had

passed through or until the filter clogged (Fig. 7.4). In the latter case, the volume filtered

was recorded. Residual water was pushed through the filter cartridge by pushing air

through it with a 60 mL sterile syringe. Cartridges were placed into whirlpack bags and

stored at -80 °C on return to the ship.

Figure 7.4 Filtration through Sterivex filter

37

Table 7.3 Locations of zodiac sampling sites.

Station Location Date and time Latitude Longitude Station

depth

(m)

Z1 - Site 1

Unnamed river (north

of Chesterfield Inlet)

05/09/2019 07:43 63° 46.139 -92° 02.470 2.5

Z1 - Site 2 05/09/2019 07:43 63° 45.961 -92° 02.004 9.2

Z1 - Site 3 05/09/2019 07:43 63° 45.760 -92° 01.601 4.2

Z1 - Site 4 05/09/2019 07:43 63° 45.347 -92° 00.971 6.3

Z1 - Site 5 05/09/2019 07:43 63° 44.840 -92° 00.816 18.0

Z2 - Site 1

Quoich River

06/09/2019 06:00 64° 07.929 -93° 39.604 15.3

Z2 - Site 2 05/09/2019 07:43 64° 05.909 -93° 38.520 16.8

Z2 - Site 3 05/09/2019 07:43 64° 04.110 -93° 36.430 30.8

Z2 - Site 4 05/09/2019 07:43 64° 02.680 -93° 33.003 38.1

Z2 - Site 5 05/09/2019 07:43 64° 00.899 -93° 30.472 23.2

Z4 - Site 1

Thelon River

08/09/2019 07:35 64° 16.061 -96° 08.486 4.5

Z4 - Site 2 05/09/2019 08:25 64° 16.476 -96° 07.611 6.5

Z4 - Site 3 05/09/2019 09:08 64° 16.624 -96° 06.494 6.0

Z4 - Site 4 05/09/2019 09:45 64° 16.840 -96° 05.402 4.6

Z4 - Site 5 05/09/2019 10:25 64° 17.826 -96° 01.923 6.1

Z6 - Site 1

Josephine River

09/09/2019 14:47 63° 01.460 -90° 39.792 1.2

Z6 - Site 2 09/09/2019 16:01 63° 01.230 -90° 38.920 3.8

Z6 - Site 3 09/09/2019 16:42 63° 01.214 -90° 38.043 7.2

Z6 - Site 4 09/09/2019 17:24 63° 01.133 -90° 36.897 9.8

Z6 - Site 5 09/09/2019 18:08 63° 01.151 -90° 35.512 13.3

Z7 - Site 1

Meliadine River

11/09/2019 07:08 62° 51.317 -92° 05.885 2.1

Z7 - Site 2 11/09/2019 08:02 62° 51.260 -92° 05.424 8.0

Z7 - Site 3 11/09/2019 08:39 62° 51.190 -92° 04.928 11.6

Z7 - Site 4 11/09/2019 09:23 62° 51.065 -92° 04.489 10.7

Z7 - Site 5 11/09/2019 10:13 62° 50.082 -92° 02.890 17.6

Z8 - Site 1

Maguse River

13/09/2019 08:02 61° 16.509 -94° 01.049 1.6

Z8 - Site 2 13/09/2019 08:53 61° 16.144 -93° 58.231 2.0

Z8 - Site 3 13/09/2019 09:32 61° 15.598 -93° 57.265 4.5

Z8 - Site 4 13/09/2019 10:09 61° 14.610 -93° 56.496 8.6

Z8 - Site 5 13/09/2019 10:46 61° 13.355 -93° 53.980 16.9

Z9 - Site 1

Tha-anne and

Thlewiaza rivers

14/09/2019 07:07 60° 29.564 -94° 38.605 1.4

Z9 - Site 2 14/09/2019 07:44 60° 28.773 -94° 37.715 2.8

Z9 - Site 3 14/09/2019 08:24 60° 28.286 -94° 36.027 3.2

Z9 - Site 4 14/09/2019 09:05 60° 28.109 -94° 30.715 15.5

Z9 - Site 5 14/09/2019 09:42 60° 27.650 -94° 27.928 16.2

38

7.2.6.2 Surface sediment collection

Samples of bulk surface sediment were collected by deploying a Ponar grab

sampler over the side of the zodiac. This was not possible at all sites due to variations in

the nature of the seabed. Surface sediment was transferred into whirlpack bags using

ethanol-sterilised palette knives (Fig. 7.5) and stored at -80 °C on return to the ship.

Figure 7.5 Surface sediment collection on board zodiac

7.2.7 Box core sampling

Surface sediment was collected at each station for which successful box cores were

obtained (Table 7.4). Approximately 1mL of surface sediment was collected into triplicate

2 mL cryovials using cut 1mL syringes (Fig. 7.6), with the intention of using these samples

for initial DNA extraction and sequencing. A further ‘bulk’ surface sediment sample was

also collected as described above for the zodiac sampling. These samples could be used

should additional material be required for the DNA analyses or if initial analyses indicate

that certain sites warrant more in-depth study. All sediment samples were stored at -80°C.

39

Figure 7.6 Surface sediment collection via syringe

Table 7.4 Box core sample locations.

Station Date and time Latitude Longitude Station

depth (m)

CH1 02/09/2019 09:49 63° 12.508 -83° 47.842 111

CH2 04/09/2019 06:51 63° 15.418 -88° 22.075 182

CII1 04/09/2019 20:01 63° 23.371 -90° 43.404 55

CII4 05/09/2019 08:41 63° 44.237 -92° 00.613 32.5

CII5 06/09/2019 08:57 63° 59.276 -93° 30.229 32.2

CII7 06/09/2019 14:14 63° 55.785 -93° 36.827 37.9

BL1 07/09/2019 08:57 64° 06.589 -94° 24.821 43.9

BL3 07/09/2019 14:56 64° 13.621 -95° 28.184 37

BL5 08/09/2019 08:30 64° 18.345 -95° 59.917 25.1

BL4 08/09/2019 13:17 64° 17.773 -95° 57.362 26.8

CII10 08/09/2019 22:16 63° 54.939 -93° 58.244 54.2

CI3 08/09/2019 14:05 63° 02.604 -90° 33.513 32.3

CI2 10/09/2019 09:32 63° 18.966 -90° 39.423 34.4

RI3 11/09/2019 07:39 62° 48.789 -92° 02.572 21.8

RI6 11/09/2019 10:42 62° 47.766 -92° 05.977 28.4

RI4 11/09/2019 14:08 62° 45.896 -92° 01.604 40.8

AV1 12/09/2019 06:30 61° 94.310 -92° 32.095 63

AV2 12/09/2019 19:09 61° 06.689 -93° 56.558 10.9

AV3 13/09/2019 09:41 61° 06.731 -94° 00.593 5.9

AV4 13/09/2019 16:43 60° 47.316 -93° 57.573 47.4

AV5 14/09/2019 08:58 60° 26.518 -94° 23.796 30.1

40

7.2.8 Microcosm setup (water)

We collected additional surface water from the rosette at selected stations and

zodiac sampling sites for setting up microcosms (summarised in Table 7.5).

Table 7.5 List of incubations prepared.

Station WAF Prep. Started WAF Completed Incubations started

CH1 02/09/2019 14:00 03/09/2019 08:00 03/09/2019

CH2 04/09/2019 14:00 05/09/2019 08:00 05/09/2019

CII4 05/09/2019 14:00 06/09/2019 08:00 06/09/2019

CII5 06/09/2019 17:30 07/09/2019 11:30 07/09/2019

BL1 07/09/2019 14:00 08/09/2019 08:00 08/09/2019

BL3 07/09/2019 22:00 08/09/2019 16:00 08/09/2019

BL5 08/09/2019 20:30 09/09/2019 14:30 09/09/2019

CII 10 09/09/2019 02:00 09/09/2019 20:00 09/09/2019

CI3 09/09/2019 22:00 10/09/2019 16:00 10/09/2019

CI1 09/09/2019 22:00 10/09/2019 16:00 10/09/2019

RI3 10/09/2019 20:00 11/09/2019 14:00 11/09/2019

Z7 Site 1 10/09/2019 20:00 11/09/2019 14:00 11/09/2019

AV1 11/09/2019 20:00 12/09/2019 14:00 12/09/2019

AV2 11/09/2019 20:00 12/09/2019 14:00 12/09/2019

Z8 Site 3 13/09/2019 02:00 13/09/2019 20:00 13/09/2019

Z8 Site 5 13/09/2019 02:00 13/09/2019 20:00 12/09/2019

*WAF = water-accommodated fraction

7.2.8.1 Preparation of a bunker fuel water-accommodated fraction

A peristaltic pump was used to pump sample water through a 0.2 μm Sterivex filter

into a sterile 500 mL glass bottle (400 mL) and a 2 L glass aspirator bottle fitted with a

glass and Teflon stopcock (1.8L). The contents of the 500 mL bottle were designated the

‘unamended filtrate’ and were stored at 4°C during preparation of the water-accommodated

fraction (WAF). A second sterile bottle was filled with 400 mL of unfiltered sample water

for use as an inoculum and also stored at 4 °C.

For preparation of the WAF, an autoclaved PTFE-coated magnetic stir bar was

added to the aspirator bottle followed by the addition of 18 mL bunker C fuel oil using a

10 mL glass Hamilton syringe. The bottle contents were stirred gently on a magnetic stirrer

to achieve a vortex extending approximately 20% of the depth of the liquid (i.e. to around

the 1.4 L mark on the bottle; Fig 7.7). This typically required stirring speeds of 120 – 140

rpm. This method aims to obtain a water phase composed of the components of the bunker

fuel at their aqueous solubilities, although microscopic suspended droplets of oil are also

believed to be present. Total petroleum hydrocarbon concentrations using this method are

typically below 10 ppm (e.g. Faksness et al., 2015). After stirring for 18 hours, covered by

aluminium foil to protect from light, the lower aqueous phase was drained through the

stopcock into three 500 mL bottles. We collected samples for hydrocarbon extraction and

analysis from each of the bottles into 40 mL glass vials. A sample of the unamended filtrate

41

was also collected in order to determine background concentrations. These samples were

stored at -20 °C.

Figure 7.7 Example of the set-up for water-accommodated fraction preparation with water

from station RI3. The aspirator bottle is being gently stirred on a magnetic stir plate. The

bottles were normally covered with foil to minimise photo-degradation but the foil was

removed to allow the photo to be taken.

7.2.8.2 Preparation of microcosms

We set up microcosms in 9 sterile 125 mL serum bottles at each station for which

WAF medium was prepared as described in Table 7.6. Water was transferred into the serum

bottles using sterile 50 mL measuring cylinders or 10 mL plastic serological pipettes. The

bottles were capped with a PTFE-lined butyl rubber stopper and crimped closed before

being incubated at 4 °C in the dark. The bottles were shaken every 24 hours to ensure they

remained well-mixed.

Table 7.6 Microcosm experiment design.

WAF medium Unamended filtrate Unfiltered

Amended 40 mL - 10 mL

Unamended - 40 mL 10 mL

Filtered 40 mL 10 mL -

7.2.9 Microcosm setup (water and sediment)

Additional microcosms were set up at two stations on the expedition. The objective

was to collect surface water samples and surface sediment samples to analyse the microbial

communities within those environments, and look at their bioremediation abilities. 2µL of

autoclaved crude oil were added to 200mL of inoculum. Surface and bottom water, and

surface sediment were collected at stations Chesterfield Inlet 1 (CI1) and Arviat 4 (AV4;

Tables 7.7 and 7.8). At each depth, three replicates were set up for three analyses: amended

surface water or surface sediment, unamended surface water or surface sediment, and killed

surface water or surface sediment. For the killed control, water or sediment was autoclaved

at 121°C for 20 minutes.

42

To begin, ~2L of surface and bottom water was collected from the Rosette into 5L

carboys, and ~500mL of sediment was collected from the box core into a sterilized Nalgene

container. Water was collected and passed through the tubing used previously. For surface

water microcosms, 2µL of crude oil was added to 200mL of surface water inoculum in

500mL bottles to attain 10ppm. For surface sediment microcosms, 2µL of crude oil was

added to 180mL of autoclaved bottom water and 20mL of surface sediment into 500mL

bottles. Bottom water was collected to ensure the sediment did not dry out during the

incubation period of the microcosms. However, this water was autoclaved to ensure it did

not interfere with the microbial activity of the surface sediment. The microcosms are stored

at 4°C for 28 days.

Table 7.7 Surface water collection for microcosm.

Station Date Latitude Longitude Depth (m) Salinity Temp (°C)

CI1 Sep 10, 2019 63°20.078 -83° 13.371 11 24.84 5.90

AV4 Sep 19, 2019 60°47.327 -93°57.505 47 28.92 7.81

Table 7.8 Surface sediment collection for microcosm.

Station Date Latitude Longitude Depth (m) Salinity Temp (°C)

CI1 Sep 10, 2019 63°20.075 -90°41.455 11.7 24.84 6.03

AV4 Sep 19, 2019 60°47.316 -93°57.573 47.4 28.92 5.81

7.3 References

Faksness, L.-G., Altin, D., Nordtug, T., Daling, P.S., and Hansen, B.H. (2015). Chemical

comparison and acute toxicity of water accommodated fraction (WAF) of source and field

collected Macondo oils from the Deepwater Horizon spill. Mar. Pollut. Bull. 91, 222–229.

8. Benthic biodiversity, biological productivity and biogeochemistry in western

Hudson’s Bay

Principal Investigator: Philippe Archambault1; Cruise participant: Camille Lavoie1

1Laboratoire d'écologie benthique, Université Laval, Pavillon Alexandre Vachon

1045 avenue de la Médecine (Québec)

43


In benthic ecosystems, the availability and quantity of food and the type of bottom

influence the distribution, abundance and richness of benthic organisms. Generally, a rocky

bottom presents a diverse assemblage of organisms (Posey and Ambrose 1994) whereas a

soft bottom is more homogenous and the presence of organisms will depend of the grain

size or of the availability of food. These types of bottoms create heterogeneity and can be

responsible for great concentrations of organisms and of the presence of individual species.

Our main sampling objective for the 2019 R/V William Kennedy expedition is to

advance biodiversity surveys of benthic communities with respect to the physical and

chemical environment. Our second objective is to investigate how the benthic food web

and organisms respond to changes in sea-ice cover and carbon input, and how these

changes could affect Arctic benthic communities and their resilience.


The box core (25cm x 25cm x 50cm) was deployed to quantitatively sample

diversity, abundance and biomass of infauna and to obtain sediment cores for sediment

analyses. From 17 successful box cores, sediments of a volume varying between 500 mL

to 4 L (depending on the sediment depth) were collected and passed through a 0.5 mm

mesh sieve and preserved in a 4 % formaldehyde solution for further identification in the

laboratory (Table 8.1). A similar sample procedure was done during 2 zodiac expeditions

at river mouths (Thelon and Josephine Rivers) where sediment was collected with a 1 L

sediment grab, brought back to the ship and sieved. At all successful stations, sub-cores of

sediments were collected for sediment pigment content (top 1 cm), organic carbon content

(top 1 cm) and sediment grain size (top 5 cm). Samples for sediment pigment were frozen

at -80°C, and all other sediment samples were frozen at -20°C. All samples were sent to

Laval University for further analyses.

At 15 stations, a benthic beam trawl (Hi-Lift Research Beam Trawl, made with 1-

1/2” 1.3mm EP web, with 3/8” and 1/4” liners; 3 m width × 1.5 m height, cod end of 2 mm

mesh size) was towed on the seabed at a speed of 2-2,5 knots for 15 minutes to survey

epibenthic species diversity, abundance, and biomass (Fig. 8.1, Table 8.2). The trawl was

monitored for duration at bottom with an RBR device in order to adjust for further surface

calculations. Catches were passed through a 2 mm mesh sieve and sorted (Fig. 8.2).

Specimens were identified to the lowest taxonomic level, then counted and weighted. The

unidentified specimens were preserved in a 4% seawater-formalin solution for further

identification in laboratory. Some specimens were preserved for collaborators (Table 8.2).

44

Figure 8.1 Benthic trawl deployment

Figure 8.2 Biota sorting

45

Table 8.1 Samples collected from the box core during Leg 4 of the 2019 R/V William Kennedy Research Cruise

*Unidentified samples were sent to Laval University.

Station Date Time Gear Latitude (N) Longitude (W) Station depth (m) Volume sieved (ml) Unidentified samples*

CHO 02/09/2019 16 :39 box core 64°06.742' 83°13.366' 14.3 ROCKY BOTTOM

CH1 03/09/2019 11:35 box core 63°12.485' 83°47.842’ 111 3000 1 x 500 mL

CH2 04/09/2019 07:15 box core 63°15.408' 88°22.119’ 182 4000 1 x

CII1 04/09/2019 20:41 box core 63°23.106' 90°42.376' 73.1 2000 1 x

CII4 05/09/2019 10:04 box core 63°44.230' 92°00.716' 38.8 4000 1 x

CII5 06/09/2019 10:39 box core 63°59'280' 93°30.283' 29.4 4000 NO

Bl1 07/09/2019 09:37 box core 64°06.548' 94°23.922' 33.3 5000 1 x 500 mL

Z_Bl_Thelon_03 08/09/2019 09:08 ponar 64°16.624’ 96°06.494’ 5.9 1000 1 x

Bl_Thelon_05 08/09/2019 10:25 ponar 64°17.826’ 96°01.923’ 16.3 1000 1 x

CII10 08/09/2019 21:40 box core 63°54.986' 93°58.025' 47.5 5000 1 x

CI3 09/09/2019 16:08 box core 63°02.630' 90°33.580' 30.8 1500 1 x

Z_CI3_Josephine 09/09/2019 NA ponar 63°01.460 90°39.792’ 1.19 0350 1 x

CI2 10/09/2019 09:20 box core 63°18.868' 90°39.849' 28.3 3000 1x

RI3 11/09/2019 07:49 box core 62°48.790' 92°02.572’ 21.8 3500 1x

RI6 11/09/2019 10:42 box core 62°47.759' 92°05.978' 28.4 4000 1 x 250 mL

AV1 12/09/2019 08:08 box core 61°44.426' 92°32.057' 64.3 3000 1 x 1 L + 1 x 500 mL

AV2 12/09/2019 19:37 box core 61°06.625' 93°56.552' 11.6 0500 1 x 200 mL

AV4 13/09/2019 16:43 box core 60°47.318' 93°57.584' 47.4 1000 1 x 1 L

AV5 14/09/2019 09:28 box core 60°26.865' 94°24.933' 27.1 1000 1 x 500 mL

46

Table 8.2 Organisms collected from the benthic trawl during Leg 4 of the 2019 R/V William Kennedy Research Cruise.

Station Date Lat

Start (N)

Lon

Start (W)

Lat

End (N)

Lon

End (W)

Depth

start

(m)

Depth

end

(m)

Species Number Biomass (g) Preserved Unidentified

samples*

CHO Sep 2

64°05.934' 83°13.988’ 64°05.383’ 83°14.518' 24.4 26.2 Argis dentata 2 13.6 STERN 1 x 500 mL

Leptoclinus maculatus NA STERN

CH1 Sep 3 63°12.498' 83°47.209' 63°12.450' 83°48.154' 112 NA Heliometra glacialis 46 1 211.0916 STERN 1 x 500 mL

Strongylocentrotus sp. 1 18.1437 STERN

Pteraster pulvillus 1 NA STERN

Gorgonocephalus eucnemis 1 63.5029 STERN

Boltenia 1 40.8233 STERN

Porifera sp. 1 108.8622 STERN

Lebbeus groenlandicus 51 244.9399 STERN

Argis dentata 19 122.4699 STERN

Sclerocrangon borealis 1 40.8233 STERN

Lebbeus polaris 23 63.5029 STERN

Spirontocaris spinus 53 95.2544 STERN

Eualus gaimardii 23 49.8951 STERN

Sabinea septemcarinata 4 NA STERN

Padalus borealis 8 58.967 STERN

Ophiura sarsi 1 NA MEYER-K.

CH2 Sep 4 63°15.503' 88°20.665' 63°15.465' 88°19.716' 181 182 Acanthostepheia 9 5 STERN 1 x 500 mL

Sabinea septemcarinata 4 18 STERN

Argis dentata 28 82 STERN

Themisto libellula 82 19 STERN

47

Unidentified fish NA NA STERN 1 x 1 L

CII4 Sep 5 63°44.450' 92°01.282' 63°44.010' 92°00.383' 39.9 NA Hormathia nodosa 11 156 STERN

Actinauge cristata 10 72 STERN

Leptoclinus maculatus 1 18 STERN

Unidentified fish 1 6 STERN

Suberites c.f. ficus 1 74 STERN

Buccinum sp. 1 NA DE COELI

KELP (5 sp.) NA NA LAVOIE

CII5 Sep 6 63°59.156' 93°29.967' 63°59.592' 93°30.283' 44.1 NA Mixocephalus octodecemspinosus 9 NA STERN NONE

BL1 Sep 7 64°06.746' 94°23.490 64°06.993' 94°24.396' 30.6 28.4 Amphipoda 5 NA STERN 1 x 250 mL

BL3 Sep 7 64°13.596' 95°28.802' 64°13.114' 95°27.074' 37.5 NA EMPTY NET

CI3 Sep 9 63°02.659 90°33.650' 63°02.753' 90°33.824' NA NA Gymnocantus tricuspis 25 NA STERN 1 x

Leptoclinus maculatus 2 NA STERN

Hiatella arctica 3 NA STERN

Lebbeus polaris 4 NA STERN

KELP (3 sp.) NA NA LAVOIE

CI2 Sep 10 63°18.924' 90°39.662 63°18.565' 90°39.866' 30.8 NA Strongylocentrotus sp. 17 319 STERN 1 x 500 mL

Leptasterias polaris 1 83 STERN

Leptoclinus maculatus 1 4 STERN

Anisarchus medius 1 4 STERN

Argis dentata 1 4 STERN

Lebbeus polaris 4 NA STERN

Gymnocantus tricuspis 3 NA STERN

Ulcina olrikii 2 NA STERN

Ophiura sarsii 1 NA MEYER-K.

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Ophiocten sericeum 4 NA STERN

RI6 Sep 11 62°47.650' 92°05.827' 62°47.713' 92°06.957' 33.2 38 Triglops murayi 1 18 STERN 1 x 500 mL

Boreogadus saida 3 7 STERN

Hyas coarctatus 1 1

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