Distended gut syndrome (DGS) in larval Atlantic cod (Gadus morhua):

towards an understanding of the underlying mechanisms of

occasional episodes of high mortality in intensive rearing systems

Yuko Kamisaka1, Ann-Elise Olderbakk Jordal1, Sandeep Garg1, 2, Erling Otterlei3 and Ivar Rønnestad1
1
Department of Biology, University of Bergen, Norway, 2Aqua Research Lab, Advanced Centre of
Zoology, University of Delhi, India (present address), 3SagaFjord Sea Farm AS, Norway

Introduction
The commercial production of Atlantic cod (Gadus morhua) has increased significantly over the last few
years, representing the most promising marine target species for aquaculture in Norway today (Gjerde et
al., 2004; Kjørsvik et al., 2004). The recent successful production of Atlantic cod juveniles in Norway is
based on intensive rearing systems with cultivated live feeds. However, one of the main remaining
problems for seed production is high mortality in the larval stage resulting in unpredictable production
outcome and economic results. Prior to the high mortality, the larvae are often characterized with low
activity, reduced appetite and reduced growth rates. Based on the visible symptoms with opaque,
distended and fluid filled gut lumen, we have termed this condition “Distended Gut Syndrome (DGS)”.

Symptoms associated with DGS

DGS has been one of the major problems in the past seasons of intensive larval cod production in Norway.
A few years ago, the syndrome affected larvae at 7-21 days post-hatch (dph) in a commercial hatchery,
but during late 2006, the critical window moved to later stages, 40-80 dph, which is well into the weaning
period for cod. The symptoms described prior to the larval mortality are similar irrespective of age, and
fits well with the DGS characteristics.
At our sampling in spring 2006 from two rearing tanks at the commercial hatchery, symptoms of
DGS were observed around 18-25 dph in one of the tanks. The larvae were characterized by low activity
followed by a reduced appetite. The gut was distended and filled with fluid (Fig. 1). The few ingested
rotifers seemed to be “bathing” in the clear gut fluid and the digestibility of ingested preys appeared to be
very low. Larvae in the other tank looked comparatively better, but total survival of juveniles was also
low (< 4%). This production case clearly shows that DGS has large impact on the economy for a
company and that it is therefore necessary to understand the underlying biological mechanisms for DGS
and digestive function in order to prevent outbreaks and increase juvenile production of Atlantic cod.
25% full 50% full 75% full

Fig. 1. Irrespective of gut fullness, Atlantic cod larvae with typical “distended gut syndrome
(DGS)” had a distended and fluid-filled gut at 25 days post-hatch (dph).

Histological examination on DGS larvae in intensive production systems

The digestive tract represents an important barrier to bacterial infection because of its large internal
surface area, the exposure to ingested feed stuffs with potential auxiliary bacteria, and also because of the
natural bacterial flora which is normally established during the early feeding stages.
Studies of dying cod larvae with DGS sampled from commercial hatchery have sometimes
shown high bacterial activity (Vibrio sp.) However, in contrast to classical acute Vibrio infection (e.g.
Listonella), the clinical patterns develops slowly and the daily mortality is low, although total
accumulative mortality is high. It is also important to emphasize that it is unknown whether bacterial
infections identified during episodes of DGS in the rearing tanks are the primary or secondary cause of
the clinic. Non-bacterial factors might be the underlying factor behind the clinic and the bacterial
problems could be secondary due to growth of bacteria in an already weakened gut. Therefore, we have
histologically examined the gut of larvae with DGS for appearance of enterocytes, signs of enteritis and
presence of bacteria in the gut.
 Most of the larvae examined by toluidine blue showed typical signs of DGS such as little food in
the anterior gut (Fig. 2B) and more food in the posterior gut (Fig. 2C). However, the epithelium looked
normal in all larvae examined. Microvilli were clearly visible and no necrosis was observed in the gut.
Parallel resin sections were also colored by gram staining, but no bacteria were identified (Figs. 2D, E).
Hematoxylin-erythrosin-saffron (HES) staining was used for histopathological examination. Gut
epithelium looked normal, and there were no signs of necrosis or bacteria, e.g. Vibrio in the gut of the
examined DGS larvae (Fig. 3A). But interestingly, necrosis was observed in kidney (renal tubule) and
ureter at 22, 25, 29 dph (Figs. 3B, C). Renal tubule also seemed extended (Fig. 3B). The same tendency
was also observed in toluidine blue stained sections.

A B C

D E

Fig. 2. The gut of DGS

affected larvae at 25 dph.
Neighbour sections were
stained in toluidine blue (B, 100µm
C) or gram staining (D, E).

A B C

100µm

Fig. 3. HES-stained larvae. A) gut of 22 dph, B) kidney of 25 dph, and C) ureter of 25 dph
larva. Arrowheads point epithelial cells which fall into the lumen.

Summary and perspective

DGS larvae showed reduced appetite and fluid filled gut, but the enterocytes of the larvae appeared
normal, and no bacteria were detected in the gut lumen. However, this does not completely eliminate the
possibility that bacteria affecting DGS might be present in the gut lumen. On the other hand, necrosis was
found in kidney and ureter. We need to study further if they are related to DGS in larval Atlantic cod. Due
to the important role of the kidney in regulation of osmo- and ion homeostasis in vertebrates including
marine teleost larvae, this warrants further studies.
The underlying biological mechanisms involved in DGS seem complex. The gut symptoms,
which might have some similarities with diarrhea, suggest an unbalance of ion- and water homeostasis in
the gut, and microbial, nutritional and physiological aspects are also possibly involved. In our further
studies, we will use microarrays to monitor the transcriptome to identify genes and pathways affected by
and possibly causing DGS. We will also use q-PCR assays to examine performance of candidate genes in
DGS affected larvae focusing on some of the key digestive processes including digestion, absorption, ion-
and water transport and immune function.

Thanks to Ms. I. U. Fiksdal and Dr. E. Karlsbakk, IMR, Norway, for assistance with histopathological evaluations.
Supported by the Research Council of Norway (projects no. 174229 and 18728) and scholarship to S. Garg.

References
Gjerde, B., Terjesen, B.F., Barr, Y., Lein, I., & Thorland, I. Aquaculture 236: 167-177 (2004).
Kjørsvik, E., Pittman, K., & Pavlov, D. In: Culture of cold-water marine fish, Moksness, E., Kjørsvik, E.,
& Olsen, Y. (ed). Blackwell Publishing Ltd, Oxford, UK (2004) pp. 204-278.