Aquaculture International 7: 81–87, 1999.

© 1999 Kluwer Academic Publishers. Printed in the Netherlands.

Effects of temperature on growth and efficiency of yolk

utilisation in eggs and pre-feeding larval stages of
Atlantic salmon

ALFREDO F. OJANGUREN*, FELIPE G. REYES-GAVILÁN† and

ROLANDO RODRÍGUEZ MUÑOZ
Departmento de Biologı́a de Organismos y Sistemas, Universidad de Oviedo 33071 Oviedo, Spain

(Received 25 June 1998; accepted 13 January 1999)

Abstract. Three embryonic stages of Atlantic salmon (Salmo salar L.) were subjected to eight constant
incubation temperatures (4, 7, 10, 12, 14, 16, 19 and 22 °C) exceeding the range usually experienced in
natural conditions. A change in thermal tolerance during the embryonic and larval development was
registered: pre-hatching stages showed an upper thermal limit at about 16 °C, while hatched larvae
survived until 22 °C. Temperature significantly affects developmental rate, resulting in a faster
development and, consequently, lower yolk weight percentage at higher temperatures. We found positive
relationships between incubation temperature and body size (length and weight) in the less developed
stages, in which some yolk remained, but size decreased at increasing temperatures when yolk was
completely exhausted.

Key words: alevin, Atlantic salmon (Salmo salar), development, egg, temperature, thermal tolerance,
yolk

Introduction

Temperature has been demonstrated to affect almost every aspect of fish early
development: hatching, emergence and initial feeding times (Alderdice and Velsen,
1978; Beacham and Murray, 1990; Blaxter, 1992; see also Heggberget and Wallace,
1984; Brännäs, 1987; Crisp, 1988; Kane, 1988; Jensen et al., 1989, for data on
Atlantic salmon, Salmo salar L.), yolk conversion efficiency (Marr, 1996; Heming,
1982; Heming and Buddington, 1988; Peterson and Martin-Robichaud, 1995; Peter-
son et al., 1996) and size and body condition at first feeding (Baynes and Howell,
1996; Peterson et al., 1977, 1996). These physiological responses reveal the
controlling effect of temperature on metabolic processes through thermal dependence
on enzymatic activity (Brett, 1970; Rombough, 1988; Blaxter, 1992). Since yolk

* To whom correspondence should be addressed at: e-mail: ojangur@sci.cpd.uniovi.es

†
Present address: Department of Animal Ecology, Umeå University, S-901 87 Umeå, Sweden
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represents the main source of energy and materials for sustaining growth in early
embryos and pre-feeding larvae (Kamler, 1992), the effect of temperature on growth
and developmental processes is well evidenced in this phase, without conflicting
demands and constraints of other organismal functions (e.g., locomotion, feeding,
social interactions).
In this study the effects of temperature on growth, development and yolk
utilisation of Atlantic salmon during the endogenous feeding period were explored,
considering (1) the effect of the incubation temperature on larval growth; and (2) the
effect of temperature on embryonic mortality and thermal tolerance.

Materials and methods

Experimental eggs and alevins were obtained from a multiparental stock. Parent fish
were obtained from Autumn-run anadromous individuals captured in three neigh-
bouring rivers of Asturias (Northern Spain; rivers Narcea, Sella and Cares) and
reared until ripe in a river-fed hatchery (Aspro; Sella basin). As efficiency in yolk
utilisation is not constant throughout development (Marr, 1966; Hodson and Blunt,
1986; Heming and Buddington, 1988), we checked temperature effects at three
different stages, defined by the relative dry weight of yolk with respect to total dry
weight (i.e. yolk 1 body dry weights). The developmental stages considered were:
non-hatched embryos with 96.2% average yolk dry weight (hereafter ‘eggs’, even
though by the end of the experimental period they had hatched at all temperatures);
recently hatched alevins, 72.9% average yolk dry weight; and more developed
alevins, 20.6% average yolk dry weight (hereafter ‘alevin 1’ and ‘alevin 2’,
respectively). For the same reason, we delimited a short experimental time (8 days),
so avoiding the integration of periods with distinct developmental processes.
Eight target temperatures were considered (4, 7, 10, 12, 14, 16, 19 and 22 °C),
including the range of usual temperatures found in rivers of northern Spain
throughout the annual cycle. Actual mean (SD) temperatures, each achieved by
placement of a chiller or heater unit connected to a thermostat, were: 3.8 (0.1), 7.4
(0.4), 10.4 (0.5), 12.1 (1.0), 13.6 (0.2), 16.0 (1.4), 19.1 (0.7), and 22.2 (1.1). Water
temperature in the hatchery during the previous 2 months (including the period from
egg fertilisation to the start of the experiment), averaged 10.2 (0.9) °C, with a range
between 8–13 °C. For 5 days eggs and alevins were taken to the experimental
temperatures by slowly warming or cooling from the hatchery temperature.
For each temperature, twenty eyed eggs, seven ‘alevin 1’ (no more fish available)
and twenty ‘alevin 2’ were maintained in darkness in different I L polyethylene
vessels with mesh-covered lateral (0.5 mm light) and bottom (0.2 mm light)
windows. Eggs and alevins were just placed on the bottom of the vessel, without any
substrate. Vessels were submerged in a 60 L plastic tank, in which water was
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continuously aerated and moved round to ensure oxygen supply and temperature
homogeneity. At the end of the experimental period all the individuals were CO2
anaesthetised, measured (to the nearest 0.01 mm) and weighed (to the nearest 0.1
mg). Before recovery from anaesthesia, alevins were fixed in 4% formalin for 1 h, to
harden yolk and allow the separation of the yolk sac from the larval body. Both
components were oven dried at 50 °C to constant weight (48 h) and then weighed
separately to the nearest 0.1 mg. To obtain initial values for all the variables
measured at the end of the experimental period, a subsample of each stage was also
measured, fixed, dried and weighed before starting the test. Relationships between
the different variables and water temperature (actual values) were fitted by the least
squares methods, using linear, logarithmic and polynomial functions.

Results

The first general result observed was a change in thermal tolerance during embryonic
and larval development. No mortality occurred at any temperature for ‘alevin 2’,
whereas mortality experienced by ‘alevin 1’ was quite high (23%), but unrelated to
temperature (regression analysis: r2 5 0.058; P , 0.05). On the contrary, survival of
non-hatched embryos proved to be strongly temperature dependent within the range
considered in this study (r2 5 0.850; P , 0.05).
Temperature had a dramatic effect on yolk absorption rate. No yolk remained at
any temperature for the initially more developed alevins, but highly significant
negative relationships with yolk dry weight percentage were found for the two less
developed stages (Fig. 1). A linear relationship between yolk dry weight percentage
and temperature was found for ‘eggs’ (non-hatched embryos), which still retained a
substantial amount of yolk at the end of the experimental period. A logarithmic
function provided the best fit to the data for ‘alevin 1’. These alevins were close to
starting to feed and yolk was nearly exhausted at the highest temperatures.
Temperature explained a very high percentage of variance in final body dry weight
for the three developmental stages (almost 90% for ‘eggs’ and more than 95% for
‘alevin 1’ and ‘alevin 2’) (Fig. 2). Body size (both length and weight, yolk excluded)
increased with temperature during stages in which yolk reserves remained at the end
of the experimental period (‘eggs’ and ‘alevin 1’), but decreased in ‘alevin 2’ as the
yolk became exhausted. For ‘eggs’, body dry weight and body length showed
positive linear relationships with temperature. For ‘alevin 1’ there was an increase in
body weight that attenuated at the highest temperatures, so that the data were well
described by a second order polynomial function. The shape was nearly the same
when considering body length, but with a slightly lower coefficient of determination.
Body size decreased with temperature for ‘alevin 2’, following a curvilinear
relationship (Fig. 2).
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Figure 1. Regressions of mean (SE) yolk weight percentage with water temperature for Atlantic salmon
‘eggs’ (open circles) and ‘alevin 1’ (black squares). The cross represents the only survivor at 19 °C in
the ‘eggs’ group; this value was not used to fit the function.

Discussion

Our data suggest the existence of an upper thermal limit at about 16 °C for advanced
eyed eggs of Atlantic salmon in these rivers. Only one out of twenty embryos
survived at 19 °C (even this embryo exhibited malformations) and none did so at 22
°C. Hatched larvae (‘alevin 1’) showed a higher upper thermal limit, they tolerated
even the highest experimental temperatures, but suffered some mortality at all
temperatures. ‘Alevin 2’ registered no mortality all along the temperature range.
Temperature seems to have different lethal effects throughout the ontogenetic
development, pre-hatching stages being more stenothermal (Peterson et al., 1977;
Rombough, 1988; Blaxter, 1992).
By using the relative amount of yolk (weight percentage) as a development
indicator, we found faster development at increasing incubation temperatures both
for ‘eggs’ and ‘alevin 1’ (Fig. 1). For ‘alevin 1’ the attenuation of the slope with
increasing temperature (Fig. 1) could result from a reduction in metabolic intensity at
the end of the endogenous feeding period (Hodson and Blunt, 1986; Kamler, 1992;
Rombough, 1994).
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Figure 2. Relationships between mean (SE) final body dry weight (BDW, black circles) and final body
length (BL, open circles), and incubation temperature for Atlantic salmon ‘alevin 2’, ‘alevin 1’ and
‘eggs’ (downward).
86

Because temperature is a critical factor in determining developmental rate (Brett,

1970; Gorodilov, 1983; Kane, 1988; Blaxter, 1992), the observed positive relation-
ships of body length and weight with temperature were the expected result. The
linear increase in body size with temperature for ‘eggs’ (Fig. 2) is attributable to the
not very advanced embryonic stage. Again the convex relationship obtained for
‘alevin 1’ (Fig. 2), can be explained as a consequence of an attenuated developmental
rate at the end of the pre-feeding phase, when the amount of yolk remaining becomes
progressively smaller (Hodson and Blunt, 1986; Kamler, 1992).
As no food was provided during the 8 days of the experiment, and ‘alevin 2’ had
completely exhausted the yolk by the end of this period, a loss in total weight as a
result of the resorption of body tissues was expected (Heming, 1982). At higher
temperatures, developmental rate is faster, metabolic costs of maintenance metabo-
lism increase and fish run out of yolk earlier (Blaxter, 1992; Rombough, 1988; 1994),
giving the negative relationship shown in Fig. 2. Moreover, alevins reared without
substrate in warmer environments have been demonstrated to decrease yolk utilisa-
tion efficiency due to higher locomotor activity (Peterson and Martin-Robichaud,
1995). However, an energetic deficit when yolk is exhausted could result in a
decrease in length, although higher metabolic costs of growth and less efficient yolk
utilisation could more likely explain smaller alevins at higher temperatures (Blaxter,
1992; Brett, 1970; Marr, 1966; Heming, 1982; Rombough, 1994).

Conclusions

1. The results show that there is a change in the effect of temperature on pre-
feeding mortality along the ontogenetic development of Atlantic salmon
embryos, earlier stages being more stenothermal.
2. Increasing temperature affects yolk absorption, accelerating developmental rate
at the three larval stages initially considered.

Acknowledgements

We thank the staff of the Aspro hatchery and the Consejerı́a de Agricultura,
Principado de Asturias for providing experimental fish and temperature data. F.
Braña provided helpful comments that greatly improved the manuscript. The
European Commission under the FAIR programme contract No. CT95–0009 sup-
ported this study.

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