Dataset: Neocalanus flemingeri lipid fullness

Name: Post-diapause Neocalanus flemingeri females morphometric measurements and calculations of lipid fullness and lipid volume taken from the R/V Sikuliaq and the R/V Tiglax in the Northern Gulf of Alaska from 2019-06-30 to 2019-09-13
Published: 2024-03-29
Keywords: Neocalanus flemingeri, reproduction, post-diapause, oceans

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Cite This Dataset

Spatial Extent: N:60.535 E:-147.8033 S:60.2783 W:-147.9867

Gulf of Alaska, sub-arctic Pacific

Temporal Extent: 2019-06-30 - 2019-09-13

Project:

Collaborative Research: Molecular profiling of the ecophysiology of dormancy induction in calanid copepods of the Northern Gulf of Alaska LTER site (Diapause preparation)

Principal Investigator:

Petra H. Lenz (University of Hawaii at Manoa, PBRC)

Scientist:

Daniel K. Hartline (University of Hawaii at Manoa, PBRC)

Kira J. Monell (University of Hawaii at Manoa, PBRC)

Student:

Kira J. Monell (University of Hawaii at Manoa, PBRC)

Contact:

Petra H. Lenz (University of Hawaii at Manoa, PBRC)

Data Manager:

Kira J. Monell (University of Hawaii at Manoa, PBRC)

BCO-DMO Data Manager:

Lynne M. Merchant (Woods Hole Oceanographic Institution, WHOI BCO-DMO)

Version:

Version Date:

2024-03-29

Restricted:

Validated:

Current State:

Preliminary and in progress

Post-diapause Neocalanus flemingeri females morphometric measurements and calculations of lipid fullness and lipid volume taken from the R/V Sikuliaq and the R/V Tiglax in the Northern Gulf of Alaska from 2019-06-30 to 2019-09-13

Abstract:

Morphometric data on Neocalanus flemingeri females were made on individuals collected in diapause in July and September, 2019 and after incubation in the laboratory for up to 4.5 weeks. Collections were made from depth in Prince William Sound in the Gulf of Alaska during two NGA LTER cruises. After sorting, females were incubated in flasks at 4-6ºC for up to 4.5 weeks. Individuals were removed for imaging and experimental incubations to study oocyte production. Morphometric analysis was used to measure prosome length and area as well as lipid sac area for lipid fullness and lipid volume calculations. Lipid consumption during diapause was minimal and modest initially.

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Methods & Sampling

Sample collection and sorting

Copepods were collected in Prince William Sound, Alaska in the summer and fall of 2019 during the Northern Gulf of Alaska Long Term Ecological Research (NGA LTER) cruises (https://nga.lternet.edu/). Females “PWS2/June” were collected on June 30th, 2019 at the sampling site PWS2 (Latitude 60° 32.1’N; Longitude 147° 48.2’W) (R/V Sikuliaq, cruise number: SKQ201915S), and the “Pleiades/September” females were collected on September 12th and 13th, 2019 at PWS2 and near the Pleiades Islands (Latitude 60° 16.7’N; Longitude 147° 59.2’W) (M/V Tiglax, cruise number: TGX201909). Copepods were collected with a Midi MultiNet (0.25 m2 mouth area; 150 μm mesh nets) towed vertically from near the bottom to the 163 surface at 0.5 m/sec (PWS2: 798 m; KIP2: 588 m). Upon retrieval, net samples were immediately diluted using filtered seawater collected from depth and kept between 4-6°C to minimize thermal stress. All females selected for the experiments were sorted under a dissecting microscope. Females were placed in groups of three into 750 mL Falcon tissue-culture flasks and incubated under dim light in an incubator for up to 4.5 weeks. Experimental emperatures were at or below deep-water temperatures in Prince William Sound (temperature settings: 4°C for June and 6°C for September). A subset of females was used in the DNA replication experiments; the remaining females were imaged for measurements of prosome length and lipid sac area (see related dataset https://www.bco-dmo.org/dataset/908514).

Experimental design

For the timeline, sets of females were removed from the incubation flasks for imaging and/or incubations in low concentrations of 5-Ethynyl-2’-deoxyuridine (EdU). Individuals that were imaged as well as incubated in EdU are identified in the dataset and provide identifications used in the related dataset.

Data Processing Description

Lipid sac and prosome length imaging

A total of 204 females were imaged for body measurements, with 31 of these females being also part of EdU experiments. Live females were placed in a chilled embryo dish with a small drop of seawater. Females were imaged laterally at 32x magnification using a Leica MZ16 microscope equipped with a 12 MPx Spot Insight camera. Using ImageJ light images were analyzed manually for three measurements: prosome length in mm, area of the lipid sac in mm2, and area of the prosome in mm2. Prosome length was measured by placing a line from the anterior to posterior tip of the prosome, measurements were rounded to the nearest 0.1 mm. Lipid sac and prosome area were measured by outlining their perimeters. Total lipid content in milligrams (mg) was estimated using the area of the lipid sac (equation: TL=0.197A1.38, where A is the lipid sac area and TL is total lipid). This relationship was established by comparing lateral images of the lipid sac to gas-chromatographic lipid measurements of three Calanus spp. that generated an equation that used lipid sac area as a proxy for lipid content. Measured areas of the lipid sac and prosome were also used to compute a lipid fullness percentage (lipid sac area/prosome sac area × 100).

A supplemental file with information such as LSID from the WoRMS taxonomy site is included for the collected species Neocalanus flemingeri.

BCO-DMO Processing

Add Latitude and Longitude columns with values from the “Methods and Sampling” section of the dataset page. Convert lat and lon to decimal degrees with 4 digit precision.

PWS2 lat 60.535 and lon -147.8033
Pleiades Lat 60.2783 and lon -147.9867

Rename column headers by replacing spaces with underscores and removing ? and () characters.

Remove units from column headers since the units will be noted in the parameters section on the dataset page.

Capitalize the first letter of each full month name in the Collection_month column.

Reformat dates from %m/%d/%Y to %Y-%m-%d

Rearrange the columns so that the Station, Latitude, Longitude, Date_collected and Collection_month columns come before experiment data.

The next columns give information about EdU incubation.

And the remaining columns give information about the imaging measurements.

More Information about this dataset

Funding Sources

Funding Source	Award Number
NSF Division of Ocean Sciences	OCE-1756767
NSF Division of Ocean Sciences	OCE-1459235
NSF Division of Ocean Sciences	OCE-1756859

Deployments

Deployment	Synonyms	Start Date	Platform	Investigator
SKQ201915S	SKQ2019-15S	2019-06-29	R/V Sikuliaq	Seth L. Danielson (Chief Scientist)	data info
TXF19	TGX201909	2019-09-11	R/V Tiglax	Russell R. Hopcroft (Chief Scientist)	data info

Instruments

Camera

Supplied Name: Spot Insight camera

Supplied Description:

12 MPx resolution

Instrument Type

Generic Name: Camera

Acronym: camera

Community Identifier: http://vocab.nerc.ac.uk/collection/L05/current/311/

Generic Description:

All types of photographic equipment including stills, video, film and digital systems.

Microscope - Optical

Supplied Name: Leica MZ16 microscope

Supplied Description:

Instrument Type

Generic Name: Microscope - Optical

Community Identifier: http://vocab.nerc.ac.uk/collection/L05/current/LAB05/

Generic Description:

Instruments that generate enlarged images of samples using the phenomena of reflection and absorption of visible light. Includes conventional and inverted instruments. Also called a "light microscope".

MultiNet

Supplied Name: Midi MultiNet

Supplied Description:

Mesh nets with 0.25 m2 mouth area and 150 μm mesh

Instrument Type

Generic Name: MultiNet

Community Identifier: http://vocab.nerc.ac.uk/collection/L22/current/NETT0099/

Generic Description:

The MultiNet© Multiple Plankton Sampler is designed as a sampling system for horizontal and vertical collections in successive water layers. Equipped with 5 or 9 net bags, the MultiNet© can be delivered in 3 sizes (apertures) : Mini (0.125 m2), Midi (0.25 m2) and Maxi (0.5 m2). The system consists of a shipboard Deck Command Unit and a stainless steel frame to which 5 (or 9) net bags are attached by means of zippers to canvas. The net bags are opened and closed by means of an arrangement of levers that are triggered by a battery powered Motor Unit. The commands for actuation of the net bags are given via single or multi-conductor cable between the Underwater Unit and the Deck Command Unit. Although horizontal collections typically use a mesh size of 300 microns, mesh sizes from 100 to 500 may also be used. Vertical collections are also common. The shipboard Deck Command Unit displays all relevant system data, including the actual operating depth of the net system.

Parameters

Date and time formats are described in python strftime() syntax.

Supplied Name	Supplied description	Supplied Units	Standard Name
Station	Station of plankton tow, stations were decided by the Northern Gulf of Alaska Long Term Ecological Research group	unitless	station
Latitude	Sampling location latitude, south is negative	decimal degrees	lat
Longitude	Sampling location longitude, west is negative	decimal degrees	lon
Date_collected	Collection date of organism Format: %Y-%m-%d	unitless	date
Collection_month	Collection month of organism. Month is the full name. Format: %B	unitless	month
EdU_sample_ID_number	Sample tube identification. If female was used in EdU experiments, then her sample ID number is listed	unitless	sample
Used_in_EdU_experiments	Yes or no question asking if female with a light microscope image was also used for EdU incubations	unitless	no_bcodmo_term
Length_of_EdU_incubation_in_hours	Length in hours of EdU incubation	hours	time_inc
Image_number_light_microscope	images were taken in a series; each image was given an identification number	unitless	sample
Date_image_was_taken	date when light microscope image was taken to be used for body size measurements Format: %Y-%m-%d	unitless	date
Time_point	experimental time point written out as hours or weeks; experiment start for body measurements was when females were collected	hours or weeks	no_bcodmo_term
Time_point_in_days	experimental time point written out as days after females were collected	days	no_bcodmo_term
Prosome_length	measured prosome length of each female	millimeters (mm)	length
Prosome_area	measured prosome area of each female	square millimeters (mm^2)	area
Lipid_area	measured lipid sac area of each female	square millimeters (mm^2)	area
Lipid_volume	lipid volume calculated as described in methods	milligrams (mg)	volume
Lipid_fullness_percentage	lipid fullness percentage of each female; lipid fullness was measured by dividing lipid area by prosome area and multiplying by100	percent (%)	no_bcodmo_term

Database

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Dataset: Neocalanus flemingeri lipid fullness