Journal of Fish Biology (2012)

doi:10.1111/j.1095-8649.2012.03344.x, available online at wileyonlinelibrary.com

The effect of temperature on embryonic and yolk-sac

larval development in the burbot Lota lota

F. Lahnsteiner*†, M. Kletzl‡ and T. Weismann§

*Department of Organismic Biology, University of Salzburg, Hellbrunnerstrasse 34, A-5020,
Austria, ‡Fishfarm Kreuzstein, Oberburgau 28, 4866 Unterach, Austria and §Bundesamt für
Wasserwirtschaft, Institut für Gewässerökologie, Fischereibiologie und Seenkunde, Scharfling
18, 5310 Mondsee, Austria

(Received 3 August 2011, Accepted 13 April 2012)

The present study investigated the effect of various temperature regimes on embryonic and yolk-sac
larval development of the burbot Lota lota. At constant temperature regimes a high percentage of
ready-to-hatch embryos, hatched larvae and normally shaped larvae was observed at the lowest
temperature (2◦ C), which significantly decreased with increasing temperatures (4 and 6◦ C). No
larvae hatched at 9◦ C. The stream temperature regime had no effect on the percentage of ready-to-
hatch embryos, but significantly decreased the percentage of hatched larvae and of normally shaped
larvae. The lake temperature regime did not affect the viability variables. Also an abrupt temperature
increase from 2 to 4◦ C after 8 days and from 2 to 9◦ C after 48 days had no effect on the evaluated
viability variables. For yolk-sac larvae no temperature related mortalities or abnormalities were
observed between 2 and 9◦ C. © 2012 The Authors
Journal of Fish Biology © 2012 The Fisheries Society of the British Isles

Key words: fish; hatching; malformation; thermal stress; yolk sac.

INTRODUCTION
The burbot Lota lota (L. 1758) is the only gadiform inhabiting fresh waters, liv-
ing in streams and lakes of Europe and North America north of 40◦ N (Van Houdt
et al., 2003; Worthington et al., 2010). It is endangered or extirpated due to habitat
loss, human activity, environmental pollution and climate change in many Euro-
pean regions (Van Houdt et al., 2003, 2005). In the alpine regions of Austria and
Bavaria efforts for restocking are being made to stabilize endangered populations
(Kainz & Gollmann, 1996). In Belgium, a L. lota reintroduction programme has
been conducted for over a decade and in the U.K. reintroduction strategies are being
developed (Worthington et al., 2008). Moreover L. lota is a candidate species for
aquaculture due to its excellent flesh quality (Wocher et al., 2011).
For conservation and culture of L. lota information about its reproductive biology
is necessary. Until now only data on semen biology (Lahnsteiner et al., 1997), semen
cryopreservation (Lahnsteiner et al., 2002; Jensen et al., 2008), and larval rearing

†Author to whom correspondence should be addressed. Tel.: +43 662 8044 5630; email: Franz.
Lahnsteiner@sbg.ac.at

1
© 2012 The Authors
Journal of Fish Biology © 2012 The Fisheries Society of the British Isles
2 F. L A H N S T E I N E R

under different food and temperature regimes were available (Kainz & Gollmann,
1996; Kujawa et al., 1999; Żarski et al., 2009). No data were available about the
effect of temperature on embryos and yolk-sac larvae. These data are, however,
of particular importance to optimize incubation conditions for artificial breeding
programmes and to judge whether the thermal regimes of specific river systems are
feasible for reintroductions.
The present study focuses on embryo and yolk-sac larval development of L. lota
in relation to temperature. Constant temperature regimes were tested to detect the
thermal limits for embryogenesis and yolk-sac larval development. Additionally, in
the case of embryos, changing temperature regimes were also tested to discriminate
temperature sensitivity of different ontogenetic stages, as well as natural temperature
regimes to assess viability at realistic environmental conditions during the reproduc-
tive period of the species. For yolk-sac larvae the survival, yolk consumption and
growth were investigated in relation to temperature.

MATERIALS AND METHODS

FISH AND EGG COLLECTION

Lota lota were caught from a wild population from Lake Mondsee, Austria (47◦ 49 N;
◦
13 24 E) in winter 2010 by net fishing and kept in the fish farm Kreuzstein in through-flow
concrete tanks supplied with stones and pots as shelter and under a natural photoperiod (47◦
N). Water supply was from Lake Mondsee (as the farm is located adjacent to this lake) at a
rate of 2–3 l s−1 . It had a conductivity of 355·0 ± 1·0 μS cm−1 at 25◦ C (mean ± s.d., three
different analyses), an acid-neutralizing capacity of 3·30 ± 0·01 mval l−1 , an O2 concentration
of 11·77 ± 0·06 mg ml−1 and a pH of 8·15 ± 0·00. Total phosphorous concentration was
0·008 ± 0·001 mg l−1 , NH+ −1 −
4 0·003 ± 0·001 mg l , and NO3 5·49 ± 0·03 mg l . Fish
−1
were fed with dead rainbow trout Oncorhynchus mykiss (Walbaum 1792) fingerlings at a rate
of c. 10% of their body mass every second day.
In 2011, fish were bred in captivity by spontaneous spawning. Fifteen males and 15 females
were transferred into a lake-water supplied spawning tank (1·5 × 2 × 0·5 m: length × width
× height) c. 3 weeks before the expected date of spawning. To prevent egg cannibalism the
tanks had a false bottom consisting of a stainless steel mesh with a mesh size of 5 mm.
The tanks were inspected daily between 0800 and 0900 hours if fish had already spawned.
Lake-water temperature was 5·09 ± 0·29◦ C from 25 December to 22 January. It decreased
to <1·5◦ C from 23 January to 30 January and remained at 1·23 ± 0·61◦ C from 1 February
to 28 February. Spawning events occurred on 14, 16, 18, 21 and 28 February indicating that
L. lota needs extended periods of cold water temperature to initiate spawning. Egg volumes
collected ranged between 250 and 600 ml, whereby 100 ml is equivalent to c. 40 000–50
000 eggs. The development stage of the eggs ranged from freshly fertilized without visible
cell cleavage to the two cell stage.

E F F E C T O F T E M P E R AT U R E O N E M B RY O S
For all embryo incubations ground water was used. It had a conductivity of 332·30 ±
2·10 μS cm−1 at 25◦ C (mean ± s.d., six different analyses), an acid-neutralizing capacity of
3·44 ± 0·05 mval l−1 , an O2 concentration of 12·96 ± 0·20 mg ml−1 and a pH of 8·05 ±
0·04. Total phosphorous and NH+ 4 were <0·01 mg l
−1
and NO−3 1·30 ± 0·04 mg l .
−1
Twelve temperature regimes were tested, which are shown in Table I and Fig. 1. For each
temperature regime there were five replicates, corresponding to the five spawning events, each
replicate consisting of 200 ± 20 fertilized eggs. Eggs were placed in purpose-made incubators,
which were hung in 50 l plastic tanks, one tank for each experimental temperature regime.

© 2012 The Authors

Journal of Fish Biology © 2012 The Fisheries Society of the British Isles, Journal of Fish Biology 2012, doi:10.1111/j.1095-8649.2012.03344.x
 E M B RY O N I C A N D L A RVA L D E V E L O P M E N T O F L O TA L O TA 3

Table I. Experimental temperature regimes for rearing Lota lota larvae

Tested temperature regimes Mean ± s.d. temperature (◦ C)

Constant temperatures
T1 1·99 ± 0·03
T2 4·00 ± 0·03
T3 6·00 ± 0·02
T4 8·99 ± 0·02
Natural temperatures
T5 3·65 ± 2·79 (stream, Fig· 1)
T6 1·72 ± 1·22 (lake, Fig. 1)
Temperature switch after 8 days (morula stage)
T7 from 2·00 ± 0·03 to 4·01 ± 0·02
T8 from 1·99 ± 0·03 to 6·01 ± 0·02
T9 from 1·99 ± 0·03 to 9·00 ± 0·03
Temperature switch after 16 days (half epiboly stage)
T10 from 2·01 ± 0·03 to 6·01 ± 0·02
T11 from 2·00 ± 0·02 to 9.00 ± 0·03
Temperature switch after 48 days (ready-to-hatch embryo stage)
T12 from 2·00 ± 0·02 to 8·99 ± 0·03

Approximately 50% of the water in the tanks was changed at seven-day intervals, and aeration
was supplied via two air-stones. Tanks were placed in a temperature controlled room set at ≤
2◦ C, and to generate the artificial temperature regimes (T1 to T4 and T7 to T12) the water
was warmed with 200 W aquarium heaters. In the changing temperature regimes, eggs were
transferred from the initial temperature to the final one without acclimatization. To generate
the two natural temperature regimes (T5 and T6) (Fig. 1) embryo incubation tanks were
placed in larger concrete tanks where stream water (T5) and lake water (T6), respectively,
were pumped through at a rate of 1 l s−1 . Under these conditions the ground water in the
incubation tanks equilibrated to the temperature of the surrounding water.

12

10
T11
Temperature (°)

4
T12
2

0
0 5 10 15 20 25 30 35 40 45 50 55 60 65
Time (days)

Fig. 1. Natural temperature regimes tested on Lota lota: stream water (T11) and lake water (T12). , Beginning
of hatching; , end of hatching.

© 2012 The Authors

Journal of Fish Biology © 2012 The Fisheries Society of the British Isles, Journal of Fish Biology 2012, doi:10.1111/j.1095-8649.2012.03344.x
4 F. L A H N S T E I N E R

Embryos and hatched larvae were observed during the experimental period under a stere-
omicroscope at ×4–10 magnification, and temperature effects were evaluated for the following
variables: per cent ready-to-hatch embryos, per cent hatched larvae, per cent normally shaped
larvae, development time until hatching and hatching period (time from the start of hatching
until all embryos had hatched).

E F F E C T O F T E M P E R AT U R E O N Y O L K - S A C L A RVA E
One-day post-hatch (dph) larvae from the first three spawning events were sampled. Larvae
of each spawning event were divided into sub-samples of 50 ± 5 larvae and stocked in 5 l
glass aquaria containing groundwater as described above. The aquaria were placed in an
acclimatized room with a temperature ≤2◦ C and warmed to 1·99 ± 0·02, 6·01 ± 0·02 and
8·99 ± 0·02◦ C (mean ± s.d.) using 25 W aquarium heaters. Larvae were not fed.
Larval survival was determined 6, 12 and 18 dph by counting the dead larvae on the bottom
of the aquarium in relation to the free swimming larvae. For morphometric investigations five
larvae were taken from each aquarium 1, 6, 12 and 18 dph and killed by prolonged exposure
to MS-222. They were transferred in a Petri dish and photographed in lateral position with a
digital camera coupled to a stereomicroscope. From the digitized micrographs the total length
(LT ), body width, head width, and minimal and maximal diameter of the yolk sac and of the
lipid vesicle were measured (Fig. 2). The size of the yolk sac and of the lipid droplet was
estimated based on the lateral area as it was not possible to photograph the larvae from their
ventral side and to measure the third axis of the yolk sac and of the lipid vesicle.

T E M P E R AT U R E M E A S U R E M E N T S
For temperature measurement eight-channel RedLab USB TC measuring units (www.
meilhaus.de) for thermo-couples with integrated cold-junction compensation were used.
Insulated Kapton thermocouples (www.novodirect.de) with non-insulated, 1·3 mm thick mea-
surement points and a reaction time of 0·5 s served as temperature sensors. The mea-
suring data were read out via a USB cable and a Windows XP supported PC using a
software self programmed with the National Instruments LabVIEW 2008 Student Version
(http://www.ni.com/labview/d/). Measuring interval was 1 min−1 . Temperature was recorded
for each embryo incubation tank and larval aquarium and for stream and lake water used to
equilibrate the temperature of the embryo incubation tanks.

S TAT I S T I C S
Data are presented as mean ± s.d. For statistical procedures percentage data were trans-
formed by angular transformation (arcsinvdata) and metrical data by log10 transformation to

2 7

3
4 5 6

Fig. 2. Morphometric landmarks measured in the yolk-sac larvae of Lota lota: total length (LT , 1), body depth
(2), maximal (3) and minimal (4) diameter of the yolk sac, maximal (5) and minimal (6) diameter of the
lipid vesicle and head depth (7).

© 2012 The Authors

Journal of Fish Biology © 2012 The Fisheries Society of the British Isles, Journal of Fish Biology 2012, doi:10.1111/j.1095-8649.2012.03344.x
 E M B RY O N I C A N D L A RVA L D E V E L O P M E N T O F L O TA L O TA 5

reach the assumptions of normal distribution. For data analysis ANOVA was used with the
embryo viability variables as dependent variables and the temperature regime as the indepen-
dent variable. In experiments where larval variables were analysed from the same samples,
but at different times, repeated-measure one-way ANOVA was used with time as the repeated-
measure variable. The Tukey’s B-test was used as a multiple comparison post hoc test to
determine which treatments differed significantly. Calculations were performed with the SPSS
16.0 statistics package (http://ibm-spss.softonic.de/).

RESULTS

E M B RY O S
At the constant temperature regimes a high percentage of ready-to-hatch embryos,
hatched larvae and normally shaped larvae was observed at the lowest temperature,
which significantly decreased with increasing temperatures (Table II). The period
of embryogenesis and the hatching period significantly decreased as temperature
increased, ranging, respectively, from 53 and 19 days at 2◦ C to 25 and 7 days at 6◦
C. No larvae hatched at 9◦ C (Table II).
At 2◦ C the percentage of ready-to-hatch embryos, the percentage of hatched
larvae and the percentage of normally shaped larvae were significantly higher than
at 4, 6 and 9◦ C (Table II). At 4 and at 6◦ C the percentage of hatched larvae and
the percentage of normally shaped larvae were significantly decreased (Table II).
Compared with the best temperature regime (2◦ C), the stream temperature regime
(T5) had no effect on the percentage of ready-to-hatch embryos, but significantly
decreased the percentage of hatched larvae and of normally shaped larvae (Table II).
On the other hand, the lake temperature regime (T6) did not affect the viability
variables, but decreased the hatching period (Table II).
Changing temperature regimes were compared to the best constant temperature
regime (T1) to evaluate the effect of temperature on specific embryonic stages. An
abrupt temperature increase from 2 to 4◦ C after 8 days (T7) and from 2 to 9◦ C after

Table II. Effect of constant (T1–T4) and natural (T5–T6) temperature regimes on the per
cent ready-to-hatch embryos, per cent hatched larvae, per cent normally shaped larvae, the
development time until hatching and the hatching period (time from the start of hatching
until all embryos had hatched) of Lota lota. Data are mean ± s.d. (n = 5), those in the same
column superscripted by different lowercase letters are significantly different (P < 0·05)

Temperature Ready-to-hatch Hatched Normal shaped Development Hatching

regime embryos (%) larvae (%) larvae (%) time (days) period (days)
Constant temperature regimes
T1, 2◦ C 92·6 ± 3·0a 90·5 ± 5·7a 88·1 ± 3·0a 53 ± 3a 19 ± 2a
◦
T2, 4 C 52·9 ± 21·3b 41·0 ± 16·4b 23·7 ± 16·0b 33 ± 2b 11 ± 1b
T3, 6◦ C 40·5 ± 24·0c 25·9 ± 8·2c 9·3 ± 12·3c 25 ± 2c 7 ± 1c
◦
T4, 9 C 4·3 ± 2·3d 0·0 ± 0·0d 0·0 ± 0·0d 19 ± 1d —
Natural temperature regimes
T5, stream 94·5 ± 4·2a 76·9 ± 8·9e 67·1 ± 7·4e 48 ± 3e 10 ± 1b
T6, lake 93·5 ± 5·4a 91·2 ± 6·5a 89·9 ± 4·7a 55 ± 3a 13 ± 1d

© 2012 The Authors

Journal of Fish Biology © 2012 The Fisheries Society of the British Isles, Journal of Fish Biology 2012, doi:10.1111/j.1095-8649.2012.03344.x
6 F. L A H N S T E I N E R

Table III. Effect of abrupt temperature changes (T5–T10) on the per cent ready-to-
hatch embryos, per cent hatched larvae, per cent normally shaped larvae, the develop-
ment time until hatching and the hatching period (time from the start of hatching until
all embryos had hatched) of Lota lota. Viability data are mean ± s.d. (n = 5), those
in the same column superscripted by different lowercase letters are significantly different
(P < 0·05)

Temperature Ready-to-hatch Hatched Normal shaped Development Hatching

regimes embryos (%) larvae (%) larvae (%) time (days) period (days)
Control
T1, 2◦ C 93 ± 3a 91 ± 6a 88 ± 3a 53 ± 3a 19 ± 2a
Temperature switch after 8 days (morula stage)
T7, 2 → 4◦ C 92 ± 7a 89 ± 8a,b 85 ± 7a 41 ± 3b 12 ± 1b
T8, 2 → 6◦ C 91 ± 7a,b 66 ± 7c 22 ± 8b 30 ± 2c 7 ± 1c
T9, 2 → 9◦ C 63 ± 11c 13 ± 10d 2 ± 2c 25 ± 1d 5 ± 1d
Temperature switch after 16 days (half epiboly stage)
T10, 2 → 6◦ C 86 ± 10b 81 ± 9b,e 15 ± 11b 35 ± 3e 7 ± 1c
◦
T11, 2→ 9 C 84 ± 12 b 78 ± 8 e 13 ± 6b 29 ± 2c 4 ± 1d
Temperature switch after 48 days (ready-to-hatch embryo stage)
T12, 2→ 9◦ C 91 ± 7a,b 88 ± 7a,b 84 ± 5a 55 ± 3a 5 ± 1d

48 days (T12) had no effect on the evaluated viability variables (Table II). An abrupt
temperature increase from 2 to 9◦ C after 8 days (T9), from 2 to 6◦ C after 16 days
(T10), and from 2 to 9◦ C after 16 days (T11) significantly reduced the percentage
of ready-to-hatch embryos, the percentage of hatched larvae and the percentage of
normally shaped larvae (Table III). An abrupt temperature increase from 2 to 6◦ C
after 8 days (T8) had no effect on the percentage of ready-to-hatch embryos, but
reduced the percentage of hatched larvae and of normally shaped larvae (Table III).
Misshaped larvae revealed either strongly curved vertebral columns, or abnormally
bent tails. These larvae were unable to swim to the water surface to inflate the
swimbladder by ingesting air and died.

Y O L K - S A C L A RVA E
Irrespective of the temperature regime, the survival rate of larvae decreased slightly
from 6 to 12 dph and sharply from 12 to 18 dph due to depletion of internal yolk
reserves [Fig. 3(a)]. After 18 dph it tended to be lower at 9◦ C than at 2 and 6◦
C, however, this decrease was not statistically significant. The larval LT and the
larval head width were similar for all temperature regimes and during the whole
investigation period. Therefore, data from all sampling dates and aquaria were pooled,
resulting in a LT of 4·55 ± 0·23 mm (n = 180) and a body depth of 0·73 ± 0·04 mm
(n = 180). The larval body depth decreased significantly from 1 to 6 dph [Fig. 3(b)],
but remained unchanged thereafter. The size of the yolk sac significantly reduced
during the investigation period [Fig. 3(c)]. At 12 dph it was significantly smaller
at 9 than at 2◦ C. Also the size of the lipid droplet decreased significantly during
the investigation period. No temperature related differences, however, were observed
[Fig. 3(d)].

© 2012 The Authors

Journal of Fish Biology © 2012 The Fisheries Society of the British Isles, Journal of Fish Biology 2012, doi:10.1111/j.1095-8649.2012.03344.x
 E M B RY O N I C A N D L A RVA L D E V E L O P M E N T O F L O TA L O TA 7

(a) (b) 0·5

a a a a,b a a a
100 a,b a,b
b
b
b 0·4

Larvae body depth (mm)

80
Larvae survival (%)

b b
b b
0·3
60 b b b
b b
40 0·2
c
c
20 c 0·1

0 0·0
1 6 12 18 1 6 12 18

(c) 4·0 (d) 1·2

3·5 aa a a a a
1·0
Larvae lipid droplet area (mm2)
Larvae yolk sac area (mm2)

3·0
0·8
2·5

2·0 0·6 b b b,c

1·5 b b b
b,c
0·4 c
1·0 b c
b,c
c 0·2
0·5 d
d d d d d
0·0 0·0
1 6 12 18 1 6 12 18
Time (dph)

Fig. 3. Effect of temperature [2 ( ), 6 ( ) and 9 ( ) ◦ C] on yolk-sac larvae of Lota lota: ± (a) larvae survival
(n = 150), (b) body depth (n = 15), (c) larvae yolk sac lateral area (n = 15) and (d) larvae lipid droplet
lateral area (n = 15). dph, days post hatch. Values are mean ± s.d.

DISCUSSION
The present study is the first to define the thermal optimum and the upper thermal
limits for embryos and yolk-sac larvae of L. lota. The incubation experiments under
constant temperature regimes showed that the temperature optimum for embryogene-
sis is ≤2◦ C, while higher temperatures cause disturbances in development (decrease
in the rate of ready-to-hatch embryos, of hatched larvae and of normally shaped
larvae). Lower thermal limits were not defined. As embryogenesis was not disturbed
in groundwater equilibrated to lake temperature (T6) which was almost continuously
<1◦ C during the first 25 days of incubation and reached minimal values of 0·6◦
C, the lower thermal limit can be expected at 0·5◦ C or even lower. The larval
malformations caused due to elevated temperature were curved bodies and tails, or
abnormally bent tails probably due to deformities of the vertebral column. In many
teleosts heat shock during somitogenesis causes vertebral deformities (Itzkowitz

© 2012 The Authors

Journal of Fish Biology © 2012 The Fisheries Society of the British Isles, Journal of Fish Biology 2012, doi:10.1111/j.1095-8649.2012.03344.x
8 F. L A H N S T E I N E R

et al., 1983; Helvik & Walther, 1993; Wargelius et al., 2009). Atlantic salmon Salmo
salar L. 1758 embryos exposed to sub-optimal high temperature during somitoge-
nesis displayed a high prevalence of caudal vertebral column condensations and on
the molecular biological level they revealed expressional disturbance in two genes
coding for markers of skeletal development (Wargelius et al., 2009). Also newly
hatched yolk-sac larvae of green sturgeon Acipenser medirostris Ayres 1854, kept at
sub-optimal temperature, developed curved vertebral columns and deformations that
were associated with elevated heat-shock protein levels (Iwama et al., 1999; Basu
et al., 2002; Werner et al., 2007).
Experiments with abruptly changing temperature regimes were conducted to dis-
criminate the temperature sensitivity of different embryonic stages. Morula stage
embryos tolerated a temperature rise from 2 to 4◦ C (T7) while higher temperatures
(T8 and T9) resulted in a significant decrease in embryo and larval viability. Temper-
atures >4◦ C were also not tolerated in the more advanced half epiboly stage (T10
and T11). This indicates that temperatures higher than 4◦ C exceed the tolerance limit
of L. lota embryos and that embryos are from the stenothermic type with a temper-
ature optimum at ≤2◦ C. The ready-to-hatch embryos were less sensitive to thermal
stress as they tolerated a temperature rise to 9◦ C (T12). Also a natural temperature
regime typical for an alpine stream in late winter and early spring (T5) exceeded
the tolerance limit of L. lota embryos. This indicates that only river systems which
maintain cold water temperatures in early spring are suitable for reproduction of L.
lota. T6 resembles the natural temperature regime of the surface water of an alpine
lake which met the optimal temperature range of L. lota embryos.
The present experiments show that the hatching period (time from the onset to the
end of hatching) is strongly influenced by temperature. Therefore, larvae developing
from the same egg batch may differ in their development stage from newly hatched
to first feeding when hatching at low temperatures. This shift in hatching might be
an advantage in nature as newly hatched larvae from the same spawn face different
environmental conditions, which could improve offspring survival. Under hatchery
conditions, however, larvae differing in their development stages are unwanted as
they may need different food and handling systems. A transfer of the ready-to-
hatch embryos from 2 to 9◦ C (T12) may be used for shortening of the hatching
period and for synchronization of hatching in fish farms. Also in brown trout Salmo
trutta L. 1758 the hatching period decreased with increasing temperature (Ojanguren
& Braña, 2003).
Yolk-sac larvae of L. lota tolerated a much wider temperature range than the
embryos as no temperature related mortalities or abnormalities were observed between
2 and 9◦ C. From 12 to 18 dph yolk was almost completely resorbed and simulta-
neously high larval mortalities due to starving were also recorded.
This indicates that first exogenous feeding of larvae should be required soon after
12 dph. Temperature, however, had only a minor effect on yolk resorption (only at
12 dph was the size of the yolk sac smaller at 9 than at 2◦ C) and no effect on larval
mortality during starving. This could suggest that the metabolic activity of yolk-sac
larvae is similar at a temperature range of 2–9◦ C. In O. mykiss yolk-sac larvae
oxygen consumption, ammonia excretion rates and utilization of the endogenous
energy substrates were directly related to temperature (Teles & Kaushik, 1990). In
the Black Sea trout Salmo labrax Pallas 1811 (Başçrnar et al., 2005), the gilthead sea
bream Sparus aurata L. 1758 (Polo et al., 1991), and the southern hake Merluccius

© 2012 The Authors

Journal of Fish Biology © 2012 The Fisheries Society of the British Isles, Journal of Fish Biology 2012, doi:10.1111/j.1095-8649.2012.03344.x
 E M B RY O N I C A N D L A RVA L D E V E L O P M E N T O F L O TA L O TA 9

australis (Hutton 1872) (Bustos et al., 2007) yolk absorption rate increased with
increasing temperature. Also in L. lota yolk resorption rate and metabolic activity
might increase if temperature exceeds the optimal temperature range.
In summary the present study indicates that L. lota embryos have a temperature
optimum of ≤2◦ C. At the morula stage they also tolerate 4◦ C. Temperatures ≥6◦
C result in a decrease in the rate of ready-to-hatch embryos, of hatched larvae and
of normally shaped larvae. This has implications for reintroduction programmes
as river systems meeting the optimal temperature regimes have to be carefully
selected in relation to possible future temperature increases due to climate change.
For hatcheries, temperature regime T7 is the most practicable one as it uses the high-
est temperature tolerated by embryos. This has advantages as it reduces the embryo
incubation time and minimizes the effort for water cooling. To shorten the hatching
period T12 is recommended. Finally, the data from the present study are derived
from a lacustrine L. lota population, but globally there exist multiple genetic clades
(Van Houdt et al., 2003, 2005). Future studies have to be conducted to find out
whether the defined temperature optima are of general validity or whether there
exist population-specific differences and adaptations.

The investigation was funded by the Austrian Climate and Energy Fund, project number
A962989.

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