Experimental design and statistical analyses of fish growth studies

Every year, numerous studies are published that compare the effects of different factors on the growth of aquaculture fish. However, comparatively little attention has been given to the experimental designs of these studies - in how many rearing units should each treatment be replicated, how many fish should be in each tank (n) and how should the data be analysed. The reliability of the results increases with increased replication and n. In reality, however, the experimental design must strike a balance between limited resources and the reliability of the statistical analysis. A survey of recent publications in Aquaculture suggests, that most (83%) aquaculture growth studies apply each treatment in triplicates with an average of 26 fish in each tank (range: 4 to 100). The minimum difference that can reliably be detected with statistical analyses is determined by the number of replications of each treatment, n, the variance of the data and the number of treatments applied. In the present study, we accumulated information on the variance of data in aquaculture growth studies on different species to estimate the minimum detectable difference and to assist researchers in designing experiments effectively. These results suggest that the variance is similar for different aquaculture species and, therefore, the same designs (level of replication and n) are suitable for studies on different species of fish. The minimum difference (MDD) in mean body-mass of different treatment groups that can be detected in a typical aquaculture study (triplicates, 25 fish in each tank and average variance) with 80% statistical power (less than 20% chance of Type II error) is around 26% of the grand mean. Increasing the n from 25 to 100 will reduce the MDD to 19% of the grand mean, while a further increase in n will have comparatively lesser effect. Increasing replication to quadruplicates or sextuplicates (with n as 100), will further reduce the MDD to 16% and 12% of the grand mean respectively. MDD under 10% of the grand mean is only possible when fish for the experiment are selected within a narrow size range to reduce variance. Simulations were performed, where samples (experiments) were repeatedly drawn from artificial populations with identical distribution and with the same experimental design as is commonly used in growth studies. Two of the populations had dose-dependent responses to treatment while one population showed no response to treatment. The resulting data was analysed with a mixed model ANOVA and by fitting either polynomials or asymptotic models to the data. Contrary to earlier suggestions, the critical treatment (minimum treatment to generate maximum response) estimated with the ANOVA approached more closely the population responses than did the critical treatments estimated with the non-linear modelsRitrýnt tímaritPeer reviewedPre prin

Impacts of Nile tilapia cage culture on water and bottom sediment quality: The ability of an eutrophic lake to absorb and dilute perturbations

Environmentally sustainable aquaculture depends on accurate understanding of the impacts of aquaculture-derived organic matter (AOM) and the ability of aquaculture systems to absorb and dilute perturbations. To this end, the present study assessed the impacts of AOM from cage culture of Nile tilapia on the ecology of Lake Victoria, Kenya, using fish cages near Anyanga Beach in Siaya County from December 2018 to October 2019. Four locations were surveyed for organic loadings from cage cultures, located 0, 50, 150 and 500 m (as a control site) away from the cages. The cage aquaculture produced increased P and N concentrations near the cages and a decreased N:P molar ratio. These changes stimulated algal growth which, in turn, affected the water quality. The organic material accumulated on the bottom under the cages, increasing the benthic BOD (BOD, >10 mg/g), a sensitive indicator of the ecological footprint of the cage aquaculture. Further, the negative ORP observed in the benthic layer suggested anoxic bacterial metabolism, possibly causing build-up of sulphides and methane. These changes altered the abundance and composition of both limnetic and benthic communities. At the beginning of the study, 22 zoobenthic taxa existed around the cages and 18 at the reference sites. Only 3 saprophilous taxa, chiefly gastropods (Physella spp.), bivalves (Sphaerium spp.) and oligochaetes (Tubifex spp.) were present at the cage site and 17 at the reference site at the end of the culture period. The Shannon diversity index exhibited a declining tendency with the length of culture period at the cage site, signifying a negative impact of aquaculture on biodiversity. The water quality recovery after cage disturbance is rapid (<4 months), noting there was no significant difference in the water quality recorded at the cage site and the other sampling sites after a fallow period of 4 months. However, the recovery of the sediment and meiofauna was far from complete at the end of this period. Moving the cages slightly (50–100 m) away from the former location may allow the benthic communities to recover and alleviate the problem. Further, the fallowing period, particularly for the Anyanga Beach site, should be extended from four to at least 5 months to allow for the environment to recover. With the rapid increase of cage fish farming in the African Great Lakes Region and with the potential for its occurrence in other lakes, there is a need to develop regulations to guide the industry, as well as the need for continuous monitoring of the environment, in order to provide information to guide investments and ensure sustainable cage farming

Effect of rearing temperature on flesh quality in Arctic charr (Salvelinus alpinus)

Currently, no information is available on the effect of progressively decreasing temperatures on the sensory quality of Arctic charr. The objective of this study was to investigate if and how different and progressively changing rearing temperatures affect the sensory quality of cooked and raw fillets of Arctic charr. The effects of rearing temperature on the sensory quality of Arctic charr (weight range 622–888 g) were investigated. Rearing temperature was changed during the experimental period as the fish were reared at constant or decreased temperature during October to April and subsequently reared at two constant temperatures from April to August establishing six different rearing temperature combinations. Significant differences were seen in all four main sensory attributes groups, that is odour, appearance, flavour and texture. Freshness characteristics such as sweet odour and flavour, and metallic odour and flavour increased with decreasing rearing temperature, whereas musty odour and flavour decreased. Rearing temperature had a clear effect on the colour of the fillets. Lower rearing temperature increased colour intensity substantially. Softness, juiciness and tenderness increased with decreased rearing temperature. A trend was seen towards more effect from final rearing temperature than initial temperature on the sensory attributes studied. Spoilage characteristics, that is, sour and rancid odour and flavour, and off-odour and flavour were either very weak or not noticeable in all sample groups.acceptedVersio

Arctic charr farming

The Arctic charr (Salvelinus alpinus L.) is a holarctic salmonid fish species with both landlocked and anadromous populations. In Scandinavia it is mainly found in the mountain area, but it also appears in deep and large lake further south, i.e. in the Alps. It is the northernmost freshwater fish and A. charr is generally regarded as the most cold-adapted freshwater fish. A. charr has been commercially farmed since the early 90ths and today, the total production is 3000, 2300 and 700 tonnes/year in Iceland, Sweden and Norway, respectively. Both in Sweden and Iceland, there are selective breeding programs in effect and the bulk of the farming production is conducted using offspring from the national breeding programs in each country. A. charr is renowned for its ability of high growth rate at low temperature and is therefore especially suitable for farming at high latitudes and altitudes. Moreover, due to the success of the breeding programs, the charr used in farms today grow faster and mature at a larger size and age than the original wild charr. Hence, although being a fairly small industry at present, A. charr farming is predicted to grow in all three countries. This booklet summarises up-to-date knowledge on A. charr farming production cycle, from brood stock to juvenile on-growing stage (Fig. 1). It is intended to be useful for people taking their first steps in A. charr culture but also to serve as a farming manual for more experienced farm staff. Therefore, the booklet is divided in to two chapters. The first chapter (Arctic charr farming in practice) provides a straight forward introduction to each production step from brood stock to juveniles. The second chapter (theory and background) presents some details and research data on the background of specific farming practices and procedures

Aerobic swimming in intensive finfish aquaculture: applications for production, mitigation and selection

We review knowledge on applications of sustained aerobic swimming as a tool to promote productivity and welfare of farmed fish species. There has been extensive interest in whether providing active species with a current to swim against can promote growth. The results are not conclusive but the studies have varied in species, life stage, swimming speed applied, feeding regime, stocking density and other factors. Therefore, much remains to be understood about mechanisms underlying findings of ‘swimming‐enhanced growth’, in particular to demonstrate that swimming can improve feed conversion ratio and dietary protein retention under true aquaculture conditions. There has also been research into whether swimming can alleviate chronic stress, once again on a range of species and life stages. The evidence is mixed but swimming does improve recovery from acute stresses such as handling or confinement. Research into issues such as whether swimming can improve immune function and promote cognitive function is still at an early stage and should be encouraged. There is promising evidence that swimming can inhibit precocious sexual maturation in some species, so studies should be broadened to other species where precocious maturation is a problem. Swimming performance is a heritable trait and may prove a useful selection tool, especially if it is related to overall robustness. More research is required to better understand the advantages that swimming may provide to the fish farmer, in terms of production, mitigation and selection

The effect of temperature on growth performance and aerobic metabolic scope in Arctic charr, Salvelinus alpinus (L.)

In recent years, Arctic char populations in Iceland have declined and the objective of this experiment was to throw further light on these changes by examining the effect of temperature (5, 9, 13, 17, and 21 °C) on the survival, growth rate, metabolism, and physiological indices of juvenile Arctic charr (initial mean body mass 4.02 ± 0.8 g). Mortality was 60% at 21 °C while at lower temperatures it was below 5%. However, Arctic charr populations in Iceland are declining in locations where the ambient temperature is lower, suggesting that other factors may be more important in determining the abundance of the species. The optimum temperature for growth was near 14 °C. The growth rate was progressively reduced at supra-optimum temperatures with almost no growth at 21 °C. Indicators of energy reserves: condition factor, relative intestinal mass, and hepatosomatic index are all consistent with reduced feed intake at supra-optimum temperatures. The standard and maximum metabolic rate (SMR; MMR), as well as the aerobic scope for activity (AS), were maximum at 13 °C. The routine metabolic rate (RMR) increased exponentially with temperature and, at T21, it was equal to the MMR suggesting, that the RMR was limited by the MMR. Moreover, increased heart- and gill mass at 21 °C are consistent with increased stress on the cardiovascular system. These findings are in keeping with the OCLTT hypothesis that the thermal tolerance of fish is limited by the capacity of the cardiovascular system to deliver oxygen and support metabolism. Taken together, the results of this experiment suggest, that growth rate is reduced at supra-optimum temperatures because of reduced energy intake, increased metabolic demand, and limitations in the capacity of the cardiovascular system to support metabolic rate at high temperatures. At lower temperatures, growth does not appear to be limited by the AS.acceptedVersio

The effect of temperature on growth performance and aerobic metabolic scope in Arctic charr, Salvelinus alpinus (L.)

In recent years, Arctic char populations in Iceland have declined and the objective of this experiment was to throw further light on these changes by examining the effect of temperature (5, 9, 13, 17, and 21 °C) on the survival, growth rate, metabolism, and physiological indices of juvenile Arctic charr (initial mean body mass 4.02 ± 0.8 g). Mortality was 60% at 21 °C while at lower temperatures it was below 5%. However, Arctic charr populations in Iceland are declining in locations where the ambient temperature is lower, suggesting that other factors may be more important in determining the abundance of the species. The optimum temperature for growth was near 14 °C. The growth rate was progressively reduced at supra-optimum temperatures with almost no growth at 21 °C. Indicators of energy reserves: condition factor, relative intestinal mass, and hepatosomatic index are all consistent with reduced feed intake at supra-optimum temperatures. The standard and maximum metabolic rate (SMR; MMR), as well as the aerobic scope for activity (AS), were maximum at 13 °C. The routine metabolic rate (RMR) increased exponentially with temperature and, at T21, it was equal to the MMR suggesting, that the RMR was limited by the MMR. Moreover, increased heart- and gill mass at 21 °C are consistent with increased stress on the cardiovascular system. These findings are in keeping with the OCLTT hypothesis that the thermal tolerance of fish is limited by the capacity of the cardiovascular system to deliver oxygen and support metabolism. Taken together, the results of this experiment suggest, that growth rate is reduced at supra-optimum temperatures because of reduced energy intake, increased metabolic demand, and limitations in the capacity of the cardiovascular system to support metabolic rate at high temperatures. At lower temperatures, growth does not appear to be limited by the AS

The effect of CO2, total ammonia nitrogen and pH on growth of juvenile lumpfish (Cyclopterus lumpus)

The objective of this study was to examine the effects of CO2, total ammonia nitrogen (TAN: nitrogen bound as either NH3 or NH4+) and pH on the growth and survival of juvenile lumpfish in a two-month growth study. The results demonstrate the complex interactions of these water quality variables. The specific growth rate (SGR) of the lumpfish was progressively reduced with increasing [CO2] concentration above 5–10 mg·L−1. However, growth may be reduced at even lower [CO2] and the results provide no clear safe limits under which the growth of lumpfish is unaffected by CO2 concentration. At the lowest [CO2] tested (8 mg·L−1) the SGR was reduced compared with controls. Moreover, in treatments where the [CO2] concentration increased to 8 mg·L−1 as pH was artificially reduced, the SGR was similarly reduced. These results indicate that lumpfish juveniles are very sensitive to increased [CO2] concentration in water. The SGR of the lumpfish was progressively reduced as the NH4+ concentration increased while maintaining the [NH3] constant (70–80 μg·l−1). However, in these treatments the [CO2] also increased to 8–10 mg·L−1 due to changes in pH and this likely contributed to reduced growth. Moreover, due to nitrification in the rearing systems, the nitrite concentration also increased to critical levels (1.5–4.2 mg·L−1) that could affect growth. Therefore, the results do not provide clear evidence for effects of increased NH4+ on the growth of lumpfish. Taken together, the results of the experiments show that lumpfish are sensitive to perturbations in water quality and provide a benchmark for operational welfare indicators in lumpfish aquaculture