Histological approach to nutritional status of the liver and gut in

Atlantic cod larvae with and without bile salt supplementation

OCEAN 2
Anja, Ingeborg, Jan, Linn, Maria, Rushik, Sofie, Taran and Tilde
TA: Bruno
Background

● Atlantic cod larvae (Gadus morhua) have poor digestive systems.

● Earlier studies have shown a positive effect of bile salts on nutritional

absorption in adult fish.

● An abundance of glycogen indicate the status of nutrient absorption.

● Taller and bigger density of microvilli is connected to better energy

uptake.

● Better energy uptake will increase the dry weight of the larvae.
Hypothesis
Hypothesis

Supplements of bile salt in the diet of Atlantic cod larvae will increase their absorption of phospholipids

Objectives

If there is a positive correlation between bile salt supplements and energy uptake in Atlantic cod
larvae, then the cod producers might be able to increase their efficiency

Assumptions

Bile salt solubilize phospholipids making it easier to absorb for the larvae

Predictions

Increased phospholipid absorption will lead to increased growth rate, higher microvilli and a higher
degree of glycogen storage
Study design – lab set up
Treatment group
● 3 water tanks with bile salt
● 100 eggs per liter of water, 100 liter capacity tanks

Control group
● 3 water tanks without bile salt
● 100 eggs per liter of water, 100 liter capacity tanks

Regulated factors
● Photoperiod, temperature, salinity, O2-levels
● Flowsystem ensures water circulation
● Cleaning the tank
Study design – procedure
● Duration of 8 weeks
● Weekly sampling
○ 15 larvae for dry weight per tank
○ 10 larvae for histological analysis per tank
○ Capture with tube using vacuum Photo by: Hendrik H. Segers
Study design – parameters

Parameters tested
●Survival
●Standard length
●Dry weight
●Histology
→ Hepatocytes size (liver)
→ Nucleus size (liver)
→ Mitochondrial size (liver) Egerton, S., Wan, A., Murphy, K., Collins, F., Ahern,
→ Glycogen energy storage (liver) G., Sugrue, I., ... & Stanton, C. (2020). Replacing
fishmeal with plant protein in Atlantic salmon (Salmo
salar) diets by supplementation with fish protein
hydrolysate. Scientific reports, 10(1), 1-16.

→ Villi height (intestine)

→ Epithelium height (intestine)
→ Microvilli height (intestine)
Study design - Repeatability
● Training (histology)

● Routines in the lab

● Take notes!

● Keep a backup of data

● Calibrate all equipment

Statistics
Use weight as a function of length to estimate the weight of larvae that is not
weighed.

T-test

To check if mean dataset for larvae fed with bile salts is significantly different from
the dataset for larvae not fed with bile salts.

One t-test for each type of measurement.

Expected results

● 20% survival rate

● the larva fed bile supplement will
perform better
● positive correlation between size of
larvae and the tests.
 Limitations and Calculating the right amount of feed
challenges - Study for the larvae

High mortality

Hard to sample the desired amount

of larvae each time
 Reflections and Agreeing on a hypothesis, number
of tanks, number of larvae
challenges - Group work Designing the control group
Trouble finding correct statistical
methods: we settled on t-test after a
long discussion, since it is the best
fit for just two sets of data
Organizing the work and getting an
equal contribution from every group
member was surprisingly easy and
spontaneous
Sources
E. Kjørsvik,T. van der Meeren,H. Kryvi,J. Arnfinnson,P. G. Kvenseth (1990) Early development of the digestive tract of cod
larvae,Gadus morhua L., during start-feeding and starvation, Journal of Fish Biology 38, 1-15

Hamre, K. 2006. Nutrition in cod (Gadus morhua) larvae and juveniles. e ICES Journal of Marine Science, 63: 267e274.

Kjørsvik E., Olsen C., Wold P., Hoehne-Reitan K., Cahu C. L., Rainuzzo J., Olsen A. I., Øie G., Olsen Y., (2009) Comparison
of dietary phospholipids and neutral lipids on skeletal development and fatty acid composition in Atlantic cod (Gadus
morhua), Aquaculture, Volume 294, Issues 3–4, 2009, Pages 246-255, ISSN 0044-8486

Marthinsen J. L. (2018) Ontogeny of the digestive system in lumpfish (Cyclopterus lumpus L.) larvae in relation to growth
and start feeding diet. A histological and stereological approach

Puvanendran V., Brown J.A. (2002) Foraging, growth and survival of Atlantic cod larvae reared in different light intensities
and photoperiods, Aquaculture, Volume 214, Issues 1–4, ISSN 0044-8486

van der Meeren, Terje & Jørstad, Knut & Solemdal, Per & Kjesbu, Olav. (1994). Growth and survival of cod larvae (Gadus
morhua L.): Comparative enclosure studies of Northeast Arctic cod and coastal cod from western Norway. ICES Marine
Science Symposia. 198. 633-645.
 Questions?

Thanks for listening!

 I

BEING SCANNED