Metabolic Rates Differ Between Wild and Domesticated

Rainbow Trout (Onchorhynchus mykiss)

Brandon Vennell
0544911
Abstract:

The pace of life hypothesis aims to use known traits of an animal to predict other traits

associated with that animal. Recent experiments with wild and domesticated strains of rainbow

trout (Oncorhynchus mykiss) provided evidence supporting the pace of life hypothesis in this

species; wild fish having depressed growth rates, aggression, and activity in comparison to

domesticated fish. However, there have been few studies that examine potential differences in

metabolic rates between strains. This information is important for understanding differences in

energetics between strains, particularly when applying bioenergetics models. Based on pace of

life theory, it is predicted that the domesticated strain should have elevated levels of growth,

aggression, and activity relative to wild fish. We used automatic intermittent flow respirometry

to measure standard metabolic rates in both domestic and wild rainbow trout and determined

domesticated rainbow trout consume approximately 22% more oxygen when at rest than their

wild counterpart at all body sizes. This difference suggests that different bioenergetic equations

representing metabolic rates should be employed by investigators examining growth and

energetics of these two strains separately.

Introduction:

The importance of individual genotypic and phenotypic variation in relation to

behavioural and physiological processes is becoming increasingly recognized (Biro and Stamps

2010). Individuals within a population can differ enormously in their expressed life histories,

including their behaviour, metabolism, age of sexual maturation, and growth rates (Biro and

Stamps 2008, Réale et al. 2010, Jørgensen et al. 2016). The cumulative effects of these life

history traits can have a substantial impact on ecological and management practices and has

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given rise to the pace of life hypothesis (Krueger et al. 1994, Ricklefs and Wikelski 2002, Biro

and Dingemanse 2009).

The pace-of-life hypothesis operates on the assumption there is a collection of

behavioural, physiological, and life history traits associated with an animal that are intrinsically

connected (Ricklefs and Wikelski 2002, Réale et al. 2010). The pace-of-life hypothesis predicts

an animal with a short life (fast pace of life) is more likely to have an increased level of activity,

metabolism and aggressiveness while an animal with a longer life (slow pace of life) would show

lower levels of aggression, activity, and metabolism (Réale et al. 2010). Recent investigations

have demonstrated positive correlations between the metabolic rate of an organism, its activity

level, and its dominance status in support of the pace of life hypothesis (Wikelski et al. 2003,

Careau et al. 2008, Biro and Stamps 2008, SeppÄnen et al. 2009, Biro and Stamps 2010, Sloat et

al. 2014, Metcalfe et al. 2016, Laskowski et al. 2016).

Although the pace-of-life hypothesis has gained traction in recent years (Réale et al.

2010, Metcalfe et al. 2016, Andersen et al. 2017) there are some studies that have found no link

between metabolic theory and the pace of life hypothesis (Zavorka et al. 2015, Laskowski et al.

2016). Laskowski and her colleagues investigated the of behaviour of pike, Esox Lucius, in

relation to life-history and state variables. Laskowski and her colleagues determined there was no

relation between standardized behaviour and any of the life-history traits the pace-of-life

hypothesis would predict (Zavorka et al. 2015).

Metabolic rates have been studied in a wide variety of animals including birds (Mathot et

al. 2009), mammals (Gębczyński and Konarzewski 2009), fish (Chabot et al. 2016), and insects

(Ketola and Kotiaho 2010). Although the first respirometry experiments on fish were conducted

in the mid-20th century (Fry and Hart 1948), the implications of these experiments were not

3
understood until later, when scientists began to investigate the relationships between body size,

temperature, energy conversion, and growth to metabolic rate (Brown et al. 2004).

The body of literature describing fish metabolism has grown significantly since the days

of Fry and Hart with more than 135 published studies on the metabolic rates of over 60 species

of fish from 28 families (Clarke and Johnston 1999). Most of the respirometric work conducted

to date has focused on the physiological, behavioural, and genetic differences in fish and how

those traits relate to the metabolic rate but these data also relate to the pace of life hypothesis

(Rao 1968, Jönsson et al. 1998, Øverli et al. 2000, Pottinger and Carrick 2001, Barton 2002).

When compiled together, previous works and their relation to metabolic rate begin to form the

variables found in the pace of life hypothesis.

Domesticated strains of fish generally displays a bolder personality with increased

growth rates, aggression, and activity giving while wild fish display depressed growth rates,

aggression, and activity (Swain and Riddell 1990, Kindschi et al. 1991, Biro et al. 2004, 2006,

Christie et al. 2016). A popular method to evaluate the pace of life hypothesis within a species of

fish is to study a domesticated strain of fish and their wild counter-part since they differ

explicitly in their behaviour (Swain and Riddell 1990, Kindschi et al. 1991, Biro et al. 2004,

2007). Recent whole lake experiments using wild and domesticated strains of rainbow trout that

to test the hypothesis that higher growth rates would lead to increased risk-taking and lower

survival in predatory environments as the pace of life hypothesis would predict (Biro et al. 2004,

2006, 2007, Biro and Post 2008). They found that the domesticated trout displayed increased

dominance and aggressiveness in the lab and showed increased levels of activity in whole-lake

experiments which could be predicted by pace-of-life theory (Biro et al. 2004, 2006, 2007, Biro

and Post 2008). The pace of life hypothesis also predicts a fast pace of life animal, such as the

4
domesticated trout in the Biro experiments, will also have an elevated metabolic rate in

comparison to its wild, slow pace of life, counterpart.

However, Biro and his colleagues didn’t examine metabolism when they conducted their

pace-of-life experiments. Therefore, there is a gap in understanding the pace of life and

bioenergetics in this species. Therefore, this experiment aims to determine the metabolic

component to the Biro experiments by testing the hypothesis that a wild strain of rainbow trout

will have a depressed standard metabolic rate (SMR) in comparison to a domesticated strain.

Methods

Fish husbandry

Two genotypes of rainbow trout (Oncorhynchus mykiss), a mixed-sex wild diploid strain

from the Ganaraska River and a domesticated triploid strain, were obtained from the Dorion Fish

Culture Station (Dorion, Ontario, Canada). There are several naturalized strains of rainbow trout

in Ontario lakes and rivers but the Ontario Ministry of Natural Resources and Forestry

(OMNRF) has exclusively used the Ganaraska strain for stocking since the early 1970’s (Ontario

Ministry of Natural Resources and Foresty 2015). The genetic and phenotypic differences

between the Ganaraska strain and other wild strains is negligible so it is assumed the Ganaraska

strain acquired is an accurate representation of wild rainbow trout (Kerr et al. 2000).

Both strains of trout were housed at the Biology Aquatics Facility of Lakehead

University in separate tanks on the same flow-through system. All tanks were kept under

constant conditions of 11.5oC ± 1oC and fed 1.5 % of their estimated body weight with

commercial fish food pellets daily.

5
Experimental Protocol

Respirometry experiments were conducted on a total of 64 trout (22 domestic, 42 wild) at

a constant temperature of 11.5oC ± 0.5oC using automatic intermittent-flow respirometry (Loligo

Systems) to determine the standard metabolic rate (SMR, mg O2/hr) of each rainbow trout. The

cylindrical respirometry chambers were used varied accordingly with fish length. A 45 mm

respirometry chamber (diameter= 4.5 cm, length= 13.8 cm) was used to determine the SMR of

trout with a total length less than 124 mm. 62 mm respirometry chambers (diameter= 62 mm,

length= 224 mm) and 100 mm respirometry chambers (diameter= 100 mm, length= 300 mm)

were used for fish between 124-220 mm and 220-249 mm.

Prior to introduction into the respirometry chamber, individuals were fasted for 24-36

hours then lightly anesthetized to measure weight, fork length, and total length. Each fish was

then size matched to the appropriate chamber based on total length and was allowed to fully

recover before introduction into the respirometry chamber. Respirometric data was collected in

AutoResp (Loligo Systems, v. 2.2.0) for 36 hours producing one metabolic value every ten-

minutes (5-minute flush, 30 second wait, and a 4 and a half minute measure phase). The first 12

hours of data from the 36 hour collection was removed as it was considered an acclimation

period and the following 24 hours were used as the experimental data.

The respiration software was set to measure the mass specific consumption of oxygen

(mg O2/g/hr). Each ten-minute measuring interval began with a five-minute flush cycle followed

by a thirty-second wait period and a four and a half minute measure period. During the flush

cycle, oxygenated water from the holding tank was passed through the respiration chamber to

oxygenate the water inside the chamber. During the thirty-second wait period the oxygenated

water supply was shut off and recirculation within the respirometry chamber began. During the

6
measure phase, the software measured the depletion of dissolved oxygen per unit of mass. The

water inside the respirometry chamber was recirculated past a fibreoptic dissolved oxygen probe

in a closed system.

Statistical analysis

AutoResp determined a R2 values for each measure phase of the experiment to determine

the linearity of oxygen consumption. The metabolic rate values containing R2 values of less than

0.90 were removed from the dataset and the remaining metabolic rates were used to calculate the

standard metabolic rate of individual trout.

The SMR of individual rainbow trout was determined using the statistical software R (v.

3.3.2) using code created by Chabot et al. for determining the metabolic rates of fish using

intermittent flow respirometry (Chabot et al. 2016). Chabot and his colleagues recommend using

the mean lowest normal distribution of the data unless the central variance of the data is greater

than 5.4. If the central variance is greater than 5.4 Chabot and his colleagues recommend using

the 0.2 quantile. The central variance of the mean lowest normal distribution was greater than 5.4

for a large portion of the dataset and so the metabolic rates produced using the 20th percentile

(0.2 quantile) were selected (see Chabot et al. for details).

The SMR data and fish mass were log-transformed and a test for the heterogeneity of

slopes was performed. The slopes were determined not to differ significantly so analysis of

covariance (ANCOVA) was used to determine if there was a relationship between the standard

metabolic rate and the strain of the fish.

7
Results

The SMR rates of 64 rainbow trout (22 domesticated trout ranging in size from 70.85 g-

271.23 g, 42 wild trout ranging in size from 4.00 g-189.63 g) at 11.5oC were determined. To

determine if the SMR scaled differently for each strain a test for the heterogeneity of slopes was

conducted. The heterogeneity of slopes test resulted in slopes that were not significantly

different, indicating the scaling of SMR with mass is the same for each strain. The results of the

ANCOVA revealed that domesticated rainbow trout had a significantly higher SMR than the

wild strain (F(2,61)=6.758, P=0.0117 ,Fig. 1).

The domesticated and wild trout shared a common slope of 0.9397 mgO2/g/hr but the

SMR (intercepts) of the domesticated and wild trout differed significantly (Fig. 1). The

domesticated trout had an intercept of approximately 1.25 mgO2/hr more when back-transformed

from the logarithmic scale. The difference of 1.25 mgO2/hr translates to a 22.54% difference in

the oxygen consumption rates of the two strains of trout.

Figure 1. A double logarithmic plot of standard metabolic rate as a function of mass by analysis

of covariance.

8
 The growth rate differed between each of the strains. The wild strain had a growth rate of

approximately 1.28 mm day-1 while the domesticated strain had a growth rate of approximately

1.52 mm day-1 (Fig. 2).

300

250
Fork Length (mm)

200

150

100 Wild Domestic

Linear (Wild) Linear (Domestic)
50
2017-01-30

2017-02-14

2017-03-01

2017-03-16

2017-03-31

2017-04-15

2017-04-30
Date

Figure 2. Growth rate of domesticated and wild trout during experiment.

Discussion

There is a significant and meaningful elevation of SMR in domesticated rainbow trout

compared with wild rainbow trout. This finding is strongly supported by the predictions of the

pace of life hypothesis, which can be sub-divided into three major components: life history,

behaviour, and physiology. Previous data collected by Biro and his colleagues supports the life

history and behavioural aspects of the pace of life hypothesis, but no data has been generated to

support the physiological relationships that are expected in the pace of life hypothesis (Biro et al.

2004, 2007). Here, we demonstrate clearly that wild and domesticated rainbow trout have

significantly different standard metabolic rates.

The results of this study indicate domesticated rainbow trout consume approximately

22.54% more oxygen per hour at rest than their wild counterpart. Alternatively, it would take a

wild trout approximately 3 days longer than a domesticated trout to remove all the oxygen out of

9
a 1 L tank filled with water at an initial oxygen concentration of 8.5 g/L. The pace of life

hypothesis predicts that domesticated trout would conform to a fast pace of life and therefore

would have an increased metabolic rate in comparison to wild trout (Ricklefs and Wikelski 2002,

Réale et al. 2010). In this study it was found that domesticated rainbow trout had both an

increased growth rate and an increased metabolic rate. This study confirms the metabolic

relationship the pace of life hypothesis would predict based on the Biro experiments.

Body mass and metabolic rate of fishes scale linearly when plotted with a double

logarithmic regression equation (Job 1955, Beamish 1964, Clarke and Johnston 1999,

Beauregard et al. 2013, Sloat et al. 2014, Chabot et al. 2016). The slope of the regression

equation produces a scaling coefficient which varies widely by species but generally can be

approximated to 0.8 mgO2/hr for most teleost’s (Clarke and Johnston 1999). This experiment

determined the scaling coefficient of rainbow trout to be 0.9397 mgO2/g/hr, much different from

some previous studies.

Studies with other species of salmonids have also produced varying scaling factors.

Studies conducted by Beauregard et al. (2013) determined the scaling factor associated with

Atlantic salmon (Salmo salar) to be 1.02 mgO2/g/hr and Beamish (1964) determined the scaling

factor associated with brown trout (Salmo trutta) to be 0.877 mgO2/g/hr. Clarke and Johnston

(1999) performed an in-depth review of metabolic studies on 69 different species of fish from

138 different studies and determine the mean scaling factor was 0.79 mgO2/g/hr (Clarke and

Johnston 1999). In the present study the scaling factor was determined to be much greater than

the mean scaling factor and was actually closer to the scaling factor for Atlantic salmon found by

Beauregard (Beauregard et al. 2013).

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 This scaling factor determined in the present study was relatively close in proximity to

other published studies such as Rao (1968) (<35% difference under 20g), Beauregard et al.

(2013) (<30% difference under 20 g), Scott et al. (2015) (~15% difference at 3.34 g), and

Beamish (1964) (<30% error at 100 g). However, the scaling factor determined in this

experiment does vary from others; such as those done by Cutts et al. (Cutts et al. 1998), Kindshi

et al. (Kindschi et al. 1991), and Job (Job 1955).

The variation in scaling exponents between studies could be due to population-specific

effects. Recent studies have investigated population-specific effects on metabolism to determine

if the metabolic rate of the same species of fish would change based on geographic location

(SeppÄnen et al. 2009). SeppÄnen et al. (2009) found there was a significant difference in the

metabolic rates of Atlantic salmon (Salmo salar) based on their geographic location. The

difference in the metabolic scaling coefficient between this study and others could be due to a

combination of population-specific differences in metabolism and methodological variation.

In conclusion, this study supports the pace of life hypothesis prediction that domesticated

rainbow trout from the will have an elevated metabolic rate in comparison to wild trout of the

same species. This study adds to the growing body of literature that supports the pace of life

hypothesis and highlights the significant physiological differences that can arise within the same

species of fish. These physiological differences can lead to unknown scientific biases such as

assumptions made during bioenergetic modelling.

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