A comparison of regression models for

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 Aquaculture 556 (2022) 738308

Contents lists available at ScienceDirect

Aquaculture
journal homepage: www.elsevier.com/locate/aquaculture

A comparison of regression models for defining EPA + DHA requirements

using the gilthead seabream (Sparus aurata) as a model species
Sam J.S. Houston a, Vasileios Karalazos b, John Tinsley b, Douglas R. Tocher a, c,
Brett D. Glencross a, Óscar Monroig a, d, *
a
Institute of Aquaculture, Faculty of Natural Sciences, University of Stirling, Stirling FK9 4LA, UK
b
BioMar Ltd., North Shore Road, Grangemouth FK3 8UL, UK
c
Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China
d
Instituto de Acuicultura Torre de la Sal (IATS), CSIC, 12595 Ribera de Cabanes, Castellón, Spain

A R T I C L E I N F O A B S T R A C T

Keywords: Carnivorous marine fish species such as gilthead seabream (Sparus aurata) require dietary eicosapentaenoic
Gilthead seabream nutrition (EPA) and docosahexaenoic (DHA) acids for optimal growth and wellbeing. The rapid growth of global aqua­
Essential fatty acids culture, along with increased proportions of dietary oil to increase growth rate of farmed fish, has meant that the
Long-chain polyunsaturated fatty acids
supply of marine oils used in aquafeeds has become limited. The shortfall has been satisfied by using vegetable
Nutrient requirements
Non-linear modelling
oils that lack EPA and DHA and, therefore, EFA (essential fatty acid) requirements of juvenile marine fish require
reassessment. A dietary trial was carried out with gilthead seabream (~25 g) that were fed diets with six EPA +
DHA levels ranging from 0.2% - 3.2% diet as fed. For each pellet size, the biometric data (weight gain, daily
growth index and feed conversion ratio) were analysed by four different regression strategies, namely split linear,
quadratic, the Gompertz function, and the four-parameter logistic function. Over the whole experimental period
(two pellet sizes) data suggested the current published requirement (1% of diet) was low and should be increased
to at least 1.2%. However, when the first pellet size for fish of 25–80 g was considered, the apparent requirement
was at least 1.4% of diet. This demonstrated in a single trial that EFA requirement was a function of fish mass,
decreasing as the fish grows. If FCR is considered, the requirement may be as high as 2%. The suitability of
different regression models varied, as the data for the first pellet was best fit by curves but, over both pellet sizes,
the split linear fit the data best. For asymptotic models (Gompertz and four-parameter logistic functions), a novel
way of defining the requirement was presented, the “elbow” calculation as a method to bisect an asymptotic
function. Therefore, using the raw data, we illustrate how a range of regression approaches could be explored
when determining nutrient requirement estimates as no single model was an ideal fit for all response curves.

1. Introduction
For marine carnivorous fish species, eicosapentaenoic acid (EPA;
20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3) are regarded as
essential fatty acids (EFA) due to the limited ability that these species
possess for their biosynthesis from the C18 precursor, α-linolenic acid
(ALA; 18:3n-3) (Castro et al., 2016). Consequently, marine carnivorous
finfish have a dietary requirement for EPA + DHA to guarantee survival,
health and normal growth (Oliva-Teles, 2012; Tocher and Glencross,
2015; Xie et al., 2021). However, EPA and DHA are primarily found in
marine oils and, currently, fish oil (FO) is the primary source of these
EFA in aquafeeds.
An essential nutrient must be present in the animal’s diet because it
cannot be synthesised endogenously, or at a sufficient rate, to fulfil
physiological demands (Sargent et al., 2003). Animals respond to
nutrient levels and these can be plotted over a gradient of dietary supply

Abbreviations: AGR, absolute growth rate (g day− 1); DE, dietary energy; DGI, daily growth index; DHA, docosahexaenoic acid (22:6n-3); EFA, essential fatty acids;
EPA, eicosapentaenoic acid (20:5n-3); FCR, feed conversion ratio; FO, fish oil; FPL, four parameter logistic; LC-PUFA, long-chain polyunsaturated fatty acids; NRC,
National Research Council; VO, vegetable oil; WG, weight gain (g).
* Corresponding author at: Instituto de Acuicultura Torre de la Sal (IATS), CSIC, 12595 Ribera de Cabanes, Castellón, Spain.
E-mail addresses: karal@biomar.com (V. Karalazos), jtinsley@biomar.com (J. Tinsley), d.r.tocher@stir.ac.uk (D.R. Tocher), bglencross@iffo.com (B.D. Glencross),
oscar.monroig@csic.es (Ó. Monroig).

https://doi.org/10.1016/j.aquaculture.2022.738308
Received 5 October 2021; Received in revised form 25 April 2022; Accepted 26 April 2022
Available online 1 May 2022
0044-8486/© 2022 Elsevier B.V. All rights reserved.
S.J.S. Houston et al.
to give a response curve. Depending on the nutrient under study the
response curve will have different regions. The response shows negative
values when a nutrient is deficient, and is approximately zero when the
nutrient supply just balances the maintenance level (Lassiter and
Edwards, 1982). When the nutrient is in sufficient supply the response
will be positive (Bailleul et al., 2009; Hauschild et al., 2010) and this is
the level that is usually considered a minimum requirement in terms of
animal production. Further increases in the nutrient supply only elicit
small gains in the response variable until maximum performance
(plateau or asymptote) is approached (Fig. 1). These levels of response
are important when considering which models may apply best to the
nutrient and lifestage in question (Glencross, 2009). The most
commonly used response is growth, but not all nutrients will arrest
growth and, consequently, the deficient, and maintenance part of the
response may not be realised. This situation applies to EFA for juvenile
marine fish (Kalogeropoulos et al., 1992; Skalli and Robin, 2004), where
low supply leads to slower growth but does not stop it completely (Jin
et al., 2017), although other symptoms of deficiency may become
apparent (Bou et al., 2017).
It should be noted that determination of a nutrient requirement de­
fines a minimum level of nutrient supply to attain optimum/maximum
animal responses. This is different from a specification which is the di­
etary concentration chosen by a formulator to achieve the desired pro­
duction outcomes (not limited to essential nutrients) associated with the
feed (Glencross, 2009). The exact characteristics of the nutrient response
curve depend on the precise nutrient, the species, its life stage, and the
selected response (Rodehutscord and Pack, 1999). The National
Research Council (NRC) Nutrient Requirements of Fish and Shrimp
Fig. 1. The effect of nutrient level upon an animal response (a dose response
curve). Low nutrient supply indicates deficiency (Def), prevention of deficiency
indicates maintenance (Main), at the “elbow” or corner of the curve the
nutrient level can be described as optimal (Opt) where additional supply does
not elicit significant gains in the response and maximum (Max) performance,
whereby further increases in nutrient level cannot increase the response level
any further (further increases may become detrimental to the response level,
toxicity). This publication proposes that, after an experiment demonstrating
deficiency and surplus, the optimum requirement ( ) can be determined using
an “elbow” calculation. 1) A linear model is taken from the y minima to y
maxima ( ). 2) The maximum perpendicular distance between this line and
the fitted values of the relevant none-linear function is found. 3) The require­
ment ( ) is taken to be the nutrient level at this point. This represents a “point
of diminished returns”, whereby further gain in the response is small relative to
additional nutrient level. However, the “elbow” calculation is still sensitive to
experimental design and may not be appropriate for all response curves. The
figure has no units or axes and the response curve can take different forms to
what is depicted.
2
Aquaculture 556 (2022) 738308
(2011) reports that the EPA + DHA requirements for gilthead seabream
(Sparus aurata) for EPA + DHA is 0.9% of diet (dry weight) based on
studies by Kalogeropoulos et al. (1992) and Ibeas et al. (1994a, 1994b,
1997, 1996). Moreover, Ibeas et al. (1997) suggested that, in addition to
the EPA + DHA dietary level, the DHA:EPA ratio was also an important
factor, with a value of 0.5 regarded as appropriate at 0.9% EPA + DHA
dietary supply.
There are some drawbacks associated with previous studies in gilt­
head seabream that warrant further investigations into the EFA re­
quirements of this species. The studies by Ibeas et al. (1994a) and
Kalogeropoulos et al. (1992) used experimental fish less than 75 g in
weight and purified diets rather than commercial formulations. In
addition, fish growth and feed conversion achieved in the studies were
not as high as could be achieved, which is likely to cause underestima­
tion of requirements. Furthermore, the fastest growth rates in juvenile
sea bream can be realised at 24 ◦ C, which is warmer than the 18–21 ◦ C
used in previous studies. In fact, many species of fish such as Atlantic
salmon (Salmo salar), Asian seabass (Lates calcarifer), turbot (Psetta
maxima), European seabass (Dicentrarchus labrax), starry flounder
(Platichthys stellatus), Korean rockfish (Sebastes schlegeli) and red drum
(Sciaenops ocellatus), have been reported to have EPA + DHA (often
referred to as n-3 long-chain (≥C20) polyunsaturated fatty acids, LC-
PUFA) requirements of ~1% of diet (Coutteau et al., 1996; Gatesoupe
et al., 1977; Glencross et al., 2014; Lee et al., 2003; Lochmann and
Gatlin, 1993). However, other species such as yellowtail flounder
(Pleuronectes ferrugineus), Japanese flounder (Paralicthys olivaceus), sil­
ver bream (Rhabdosargus sarba) and striped jack (Pseudocaranx dentex)
have been shown to require EPA + DHA at a higher level (1.3–2.5%)
(Leu et al., 1994; Takeuchi, 1997; Takeuchi et al., 1992; Whalen et al.,
1998). Thus, EFA requirements vary among fish species, but also among
developmental stages, with larvae known to require relatively higher
levels of LC-PUFA, particularly DHA, to satisfy the high demands
required in rapidly growing neural tissues where DHA is required
(Takeuchi, 1997; Tocher, 2010).
It is appropriate to revisit the question of EPA + DHA requirements
for two key reasons. First, in the last two decades there has been a
substantial reduction in the use of fish meal (FM) and FO (sources of EPA
and DHA) in aquafeeds. Second, the oil content of modern aquafeeds has
increased to supply high energy to promote growth while sparing pro­
tein (Glencross, 2009; Sargent et al., 2003), and it has been suggested
that EFA requirement may vary with the lipid content of the diet
(Glencross and Smith, 2001; Watanabe, 1982). Therefore, the feed in­
gredients that contain EPA and DHA are being reduced while dietary
energy is increased by the addition of vegetable oils (VO) that lack LC-
PUFA, which may affect the apparent requirement. The present study
investigated the EFA requirements of a commercially important marine
species, the gilthead seabream, fed a dietary gradient of EPA + DHA
achieved by blending FO and VO. We herein discuss the benefits and
drawbacks of a range of regression models used to determine nutrient
requirements in fish (NRC, 2011), and provide estimates of EFA re­
quirements in juveniles calculated under the different regression stra­
tegies involving different response variables, including weight gain
(WG), daily growth index (DGI), and feed conversion ratio (FCR).
2. Materials and methods
2.1. Diets, fish husbandry and sampling
Gilthead seabream were maintained under the current European
legislation on handling experimental animals (2010/63/EU). In addi­
tion, all research performed by the Institute of Aquaculture, University
of Stirling (UoS) is subject to thorough ethical review carried out by the
UoS Animal Welfare and Ethical Review Board (AWERB) prior to any
work being approved. All projects, irrespective of where they are carried
out, are submitted to AWERB for approval using detailed Ethical
Approval forms that require all aspects of the experimentation to be
S.J.S. Houston et al. Aquaculture 556 (2022) 738308

described including all animal health and welfare issues as well as other Table 2
ethical considerations. The present research was assessed by the UoS Fatty acid composition of the experimental diets given as percentage of total
AWERB and passed the ethical review process. fatty acids.
Six experimental diets were formulated to deliver graded levels of Fatty acid (%) D1 D2 D3 D4 D5 D6
EPA + DHA to juvenile gilthead seabream. The diet formulations and 14:0 0.55 1.07 1.47 2.10 3.34 5.73
measured proximate compositions are detailed in Table 1 and fatty acid 16:0 15.44 15.62 15.71 16.05 16.69 17.52
compositions in Table 2. Commercially available oils including FO, 18:0 2.53 2.62 2.67 2.80 3.04 3.43
rapeseed oil and palm oil were blended to achieve specific levels of EPA 20:0 0.46 0.38 0.43 0.41 0.36 0.26
22:0 0.23 0.23 0.22 0.21 0.19 0.16
+ DHA. Other ingredients were selected to meet the known nutrient
ΣSFAa 19.35 20.04 20.64 21.70 23.75 27.22
requirements of this species (NRC, 2011; Oliva-Teles et al., 2011). Diets 16:1n-7 0.51 1.12 1.48 2.20 3.65 6.71
were produced by extrusion at the BioMar Tech-Centre (Brande, 18:1n-9 47.13 43.91 41.89 37.57 29.37 12.38
Denmark) and referred to as D1 – D6, with D1 containing only VO and 18:1n-7 2.47 2.55 2.58 2.69 2.77 2.99
D6 only FO as added oil. 20:1n 0.93 0.97 0.98 1.04 1.11 1.29
22:1n-11 0.08 0.14 0.19 0.28 0.43 0.75
The nutritional trial was carried out at the BioMar Aquaculture 24:1n-9 0.14 0.17 0.19 0.23 0.29 0.47
Technology Centre (Hirtshals, Denmark) as described previously ΣMUFAa 51.32 48.92 47.37 44.10 37.77 24.69
(Houston et al., 2017). Briefly, seabream juveniles of approximately 3 g 18:2n-6 21.42 20.42 19.69 18.56 16.16 11.22
were purchased from a commercial hatchery (Les Poissons du Soleil, 18:3n-6 0.00 0.03 0.04 0.07 0.12 0.25
20:2n-6 0.06 0.07 0.07 0.08 0.10 0.14
Balaruc-les-Bains, France) and initially fed a commercial diet containing
20:4n-6 0.05 0.11 0.15 0.23 0.40 0.78
FM and FO from first feeding until they reached ~24 g. At the start of the 22:5n-6 0.00 0.04 0.05 0.09 0.16 0.31
trial, 150 fish were randomly distributed in each of 18 × 1 m3 tanks with Σ n-6 PUFAa 21.52 20.67 20.01 19.03 17.01 12.90
each treatment being fed in triplicate (n = 3 per diet). The tanks were 18:3n-3 6.05 5.69 5.52 5.03 3.99 1.76
part of a recirculation aquaculture system with photoperiod, tempera­ 18:4n-3 0.08 0.30 0.44 0.70 1.23 2.40
20:4n-3 0.03 0.08 0.11 0.18 0.32 0.62
ture and salinity maintained at 12 L:12D, 24 ◦ C and 32 g kg − 1,
20:5n-3 0.70 1.96 2.73 4.32 7.48 14.34
respectively. The total duration of the feeding trial was 128 days, during 22:5n-3 0.11 0.26 0.35 0.53 0.91 1.71
which time the fish were fed two pellet sizes, firstly a 3 mm pellet (P1) 22:6n-3 0.63 1.47 1.98 3.06 5.23 9.89
for 56 days and, secondly a 4.5 mm pellet (P2) for 72 days. Therefore, Σ n-3 PUFAa 7.61 9.76 11.13 13.83 19.16 30.77
EPA:DHA 1.11 1.33 1.34 1.41 1.43 1.45
three experimental periods were considered, namely P1, P2 and over the
EPA + DHA (%)b 0.22 0.57 0.78 1.10 1.91 3.22
whole trial (OV), with the tank biomass being measured at 0, 56 and
a
128 days. At the end of P1 and P2, feeding was stopped 24 h before SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; PUFA,
polyunsaturated fatty acids; EPA, eicosapentaenoic acid (20:5n-3); DHA, doco­
sahexaenoic acid (22:6n-3).
b
These values are the predictor variable for all analyses in this publication,
Table 1
expressed in this table as a percentage of diet.
Diet formulations and proximate analyses of the six experimental diets.
DIET D1 D2 D3 D4 D5 D6
sampling and, prior to weighing, fish were anaesthetized with benzo­
Ingredients (% of diet as caine (Centrovet, Kalagin, Santiago, Brazil), recovered from the anaes­
fed) thetic bath to a table to allow excess water to drain away, fish weighed
Fishmeal 12.50 12.50 12.50 12.50 12.50 12.50 and biomass from each of the tanks recorded and fish counted. The fish
Soy protein concentrate 21.90 21.90 21.90 21.90 21.90 21.90 were fed twice daily ad libitum and the delivered and wasted feed
Rape seed meal 10.00 10.00 10.00 10.00 10.00 10.00
recorded for an accurate determination of biological FCR.
Wheat gluten 4.00 4.00 4.00 4.00 4.00 4.00
Corn gluten 25.00 25.00 25.00 25.00 25.00 25.00
Wheat 7.10 7.10 7.10 7.10 7.10 7.10
2.2. Requirement analysis
Lysine 0.70 0.70 0.70 0.70 0.70 0.70
Methionine 0.07 0.07 0.07 0.07 0.07 0.07
Vitamin and mineral Although EPA and DHA are individual nutrients, they are supplied
0.45 0.45 0.45 0.45 0.45 0.45
premixa together in marine oils used by the aquafeed sector and, thus, in the
Microingredientsb 2.67 2.75 2.79 2.88 3.07 3.40 present study, were analysed as though they represent a single nutrient.
Yttrium 0.03 0.03 0.03 0.03 0.03 0.03
Therefore, the predictor variable in the following analysis was EPA +
DHA% of diet (as fed). Three related response variables were selected for
Oils (%)
analysis, weight gain (WG = final weight – initial weight), daily growth
Fish oil (SA) 0.00 1.75 2.64 4.43 8.00 14.85
Rapeseed oil 10.40 9.20 8.60 7.30 4.80 0.00 index (DGI = (final weight0.333 – initial weight0.333) / days ⋅ 100) and
Palm oil 5.19 4.56 4.22 3.64 2.39 0.00 food conversion ratio (FCR = (final weight – initial weight) / feed
consumed). To estimate the requirements of gilthead seabream juveniles
Proximate composition (% of diet as fed) for EPA + DHA% of diet, a series of regression analyses were performed
Crude protein 41.8 43.0 42.9 41.7 42.4 42.5 on the three responses WG (g), DGI and FCR, for the P1 (3 mm), P2 (4.5
Crude lipid 22.2 21.7 21.7 21.5 21.8 20.8 mm) and OV (both pellets) response data. All regression analyses were
Ash 6.1 6.1 5.9 5.9 6.1 6.2
undertaken using the statistical package R (R Core Team, 2013, version
Moisture 10.3 9.7 9.4 10.7 8.9 8.9
Crude energy (MJ/kg) c
22.0 22.1 22.2 21.8 22.2 22.0
3.4.0; Vienna, Austria). The model parameters were obtained using the
a
function nls(), and the following “packages” to analyse the data: nlstools
Vitamin and mineral premix including (as IU or ppm of premix): Vitamins A (Baty et al., 2015), propagate (Speiss, 2013) and ggplot2 (Wickham,
(666,666 IU/kg), E (40,000 ppm), K3 (4002 ppm), B1 (4000 ppm), B2 (4800
2016). The package nlstools contains a range of functions for performing
ppm), B3 (12,000 ppm), B5 (16,000 ppm), B6 (4000 ppm), B12 (4 ppm), Biotin
regression diagnostics, while propagate’s function (predictNLS()) was
(120 ppm), Folic acid (4000 ppm), C (28,063 ppm), Copper (222 ppm), Cobalt
(222 ppm), Iodine (133 ppm), Manganese (1784 ppm), Zinc (22,223 ppm). used to calculate the prediction intervals for a model’s fitted values (by
b
Monocalcium-phosphate (MCP) (1.62% diet), cholesterol (0.08–0.17 diet), Monte-Carlo simulation and second order Taylor expansion), with
Emulthin G35 (0.6–1.4% diet), antioxidants (0.03% diet). graphics produced in ggplot2. There are many types of non-linear
c
Estimated by using the mean values of gross energy for proteins, lipids and functions that can model these experimental data and the approach
carbohydrates 23.6, 39.5 and 17.2 kJ/g, respectively. presented was empirical, as opposed to mechanistic. Preference was

3
S.J.S. Houston et al. Aquaculture 556 (2022) 738308

given to models with low error (mean squared error, MSE), and that
were a strong fit to the requirement region of the response. Suitable
models were assessed diagnostically according to techniques described
by Ritz and Streibig (2008). The four regression models used were: split
linear model, split quadratic model, Gompertz model and four-
parameter logistic (FPL) model. For the latter two models (asymptotic
models) two requirement estimates are presented: 1) 95% of the
asymptote (NRC, 2011), which is referred to as the “NRC criterion” and
plotted as ▴in the relevant figures and, 2) the “elbow” calculation,
plotted as ● in the relevant figures. The elbow calculation is a graphical
method that takes the maximum and minimum response values of a
model, joins these with a straight line, then defines the point of the
model’s curve which has the furthest perpendicular distance from this
line (see Fig. 1). The elbow’s usual application is in K-means clustering
to select an appropriate number of clusters within unstructured datasets
(Syakur et al., 2018). The R-code used was adapted from a stackerflow
post (Eickhart, 2017).
Unless otherwise stated n = 18, which in regression is sufficient to
detect large (f2 = 0.5) effects in the response.
2.2.1. Split linear model
The first model applied to the responses was the so-called “broken
line” or split regression model, in which the first segment is a linear
model usually ascending (descending for FCR) to a maximum response,
i.e. a horizontal segment (a mean). The requirement is defined as the
intersection of these two segments. Split linear regression can be
expressed as follows:
f(x) = m ⋅ x + c for x < req
f(x) = m ⋅ req + c for x > req
where x is the percentage of EPA + DHA in the experimental diet, c is
the model’s intercept and m is the gradient (of the first segment) and req
is x at the breakpoint (the requirement).
2.2.2. Split quadratic model
The second model applied to the data was the split quadratic model,
in which the first segment is a quadratic function ascending to a
maximum level of response, whereas the second segment is horizontal.
The requirement is defined as the intersection of these two segments.
Split quadratic regression can be expressed as follows:
f(x) = m ⋅ x + (− 0.5 ⋅ m / req) ⋅ x2 + c for x < req
f(x) = m ⋅ req + (− 0.5⋅m / req) ⋅ req2 + c for x > req
where x is the percentage of EPA + DHA in the experimental diet, c is
the model’s intercept, m is the rate constant and req is x at the breakpoint
(the requirement).
2.2.3. Gompertz model
The first asymptotic function fitted to the response data was the
Gompertz that can be defined as:
f(x) = a ⋅ exp. (b ⋅ c x)
where x is the percentage of EPA + DHA in the experimental diet, a is
the asymptote or Y maxima, b sets the displacement on the x-axis and c is
the scaling parameter, setting the rate of growth towards the asymptote.
2.2.4. Four parameter logistic model
The second asymptotic function fitted to the response was the FPL
model that is defined as:
f(x) = b + (a-b) / (1 + exp.((xmid-x)/c))
where x is the percentage of EPA + DHA in the experimental diet, a is
the upper asymptote, b is the lower asymptote, xmid is the value of x at
the inflection point and c is the scaling parameter.
3. Results
3.1. Growth performance
Table 3 shows the mean initial weights, final weights, DGI and FCR.
During the feeding trial, weight of fish in all dietary groups increased
(Table 3). However, the increase in dietary EPA + DHA (particularly
diets D5-D6) resulted in higher weight gain in gilthead seabream
juveniles.
3.2. Requirement analysis
A series of regression analyses were conducted to explore different
ways to determine the EPA + DHA requirement of juvenile gilthead
seabream. All functions and their respective parameters are given in
Table 4. The requirement estimates derived from these models are given
in Table 5, along with measures of the model fit, the Akaike information
criterion (AIC) and the MSE. Below we present the EFA requirement esti­
mations resulting from applying the four models to growth parameters (WG
and DGI) and FCR.
3.2.1. Weight gain and daily growth index
Using the split linear model to fit the WG data, the requirements were
estimated as 1.21, 1.47 and 1.13% EPA + DHA as a percentage of diet for
OV, P1 and P2, respectively (Fig. 2A). Using the split linear model to fit
the DGI data, the requirements were 1.20, 1.46 and 1.09% EPA + DHA
for OV, P1 and P2, respectively (Fig. 2B). The requirement values agree
between the two metrics of growth, WG and DGI. The distribution of
error for P1, WG and DGI exhibited a curve. When using the split
quadratic model for the WG data, the EFA requirements were estimated
as 1.70, 2.25 and 1.57% EPA + DHA for OV, P1 and P2, respectively
(Fig. 2C). The error distribution in the P1 data was better using the
quadratic model. Using the split quadratic model for the DGI data, the
requirement levels were 1.64 and 2.15% EPA + DHA for OV and P1,
respectively. This model failed to fit the DGI data of P2 due to the high
DGI values attained with fish fed diet D4 (Fig. 2D). The OV and P2 data
was best modelled by the split linear method, but the P1 data was best
described by the split quadratic model (Table 5).
The asymptotic non-linear models, namely the Gompertz and the
four-parameter logistic (FPL) models, applied to the WG and DGI data
are presented in Fig. 3. Using the Gompertz model and elbow calculation
(Fig. 1) to fit the WG data, the EFA requirements were estimated as 1.22,
1.34 and 1.18% EPA + DHA for OV, P1 and P2, respectively (Fig. 3A).
Considering the NRC recommendation of 95% of the maximum
response, the EFA requirements were calculated as 1.13, 1.40 and 1.07%
EPA + DHA for OV, P1 and P2, respectively. With regards to the DGI

Table 3
Mean values (± SD) for the dietary treatments for the biometric data analysed in this study (n = 3).
D1 SD D2 SD D3 SD D4 SD D5 SD D6 SD

Initial weight (g) 24.4 ±0.3 24.3 ±0.1 24.0 ±0.1 24.6 ±0.3 24.3 ±0.1 24.5 ±0.3
Final weight (g) 198.6 ±1.3 219.4 ±6.6 224.8 ±5.0 241.3 ±3.1 245.0 ±4.0 248.3 ±1.5
Weight gain (g) 174.2 ±1.3 195.1 ±6.6 200.8 ±4.9 216.7 ±2.8 220.7 ±3.9 223.8 ±1.4
DGIa 2.29 ±0.01 2.44 ±0.05 2.49 ±0.04 2.58 ±0.01 2.62 ±0.02 2.63 ±0.01
FCRb 1.33 ±0.02 1.33 ±0.02 1.31 ±0.02 1.27 ±0.01 1.22 ±0.01 1.21 ±0.01
a
Daily Growth Index = Final weight (g) 0.333 – Initial weight (g) 0.333/Days feeding.
b
Food conversion ratio = (Feed fed (kg) – feed waste (kg))/Δ Biomass (kg).

4
S.J.S. Houston et al. Aquaculture 556 (2022) 738308

Table 4
Model parameters fitted to the biometric data of S. aurata juveniles in the present trial to derive estimates for the EPA + DHA requirement of this species. The four
models presented are the split linear, split quadratic, Gompertz and the four parameter logistic (FPL) function. The models were applied to the overall data (OV, both
pellet sizes), the 3 mm pellet size (P1, in the text) and the 4.5 mm pellet size (P2 in the text). The name of the parameter and the notation used in the text is indicated in
the column headers. n = 18 except for the Gompertz model applied to FCR data, where n = 15.
Model type Parameters

Metric Period Plateau Gradient Intercept Requirement

m c
Split regression WG (g) OV 222.22 47.13 165.25 1.21
3 mm 78.81 11.02 57.61 1.47
4.5 mm 148.38 36.03 107.62 1.13
DGI OV 2.63 0.33 2.23 1.20
3 mm 3.05 0.32 2.59 1.46
4.5 mm 2.30 0.32 1.96 1.09
FCR OV 1.21 − 0.07 1.36 2.03
3 mm 1.09 − 0.06 1.23 2.50
4.5 mm 1.25 − 0.10 1.41 1.62

Plateau Rate constant Intercept Requirement

m c req
Ascending polynomial WG (g) OV 222.43 75.26 158.52 1.70
3 mm 74.04 15.62 56.43 2.25
4.5 mm 148.76 59.18 102.31 1.57
DGI OV 2.63 0.55 2.17 1.64
3 mm 3.06 0.48 2.54 2.15
FCR OV 1.21 − 0.11 1.37 3.03
3 mm 1.02 − 0.06 1.23 6.94
4.5 mm 1.24 − 0.13 1.42 2.71

Plateau Displacement Scale parameter

a b c
Gompertz WG (g) OV 224.42 − 0.38 0.17
3 mm 72.84 − 0.31 0.28
4.5 mm 149.99 − 0.42 0.14
DGI OV 2.64 − 0.22 0.16
3 mm 3.08 − 0.21 0.26
4.5 mm 2.31 − 0.23 0.10
FCR OV 1.20 0.23 0.26
3 mm 1.06 0.21 0.47
4.5 mm 1.24 0.26 0.20

Plateau Bottom Inflection Scale parameter

a b xmid c
FPL WG (g) OV 223.00 150.65 0.45 0.32
3 mm 74.62 24.75 − 0.32 0.66
4.5 mm 149.00 106.37 0.55 0.25
DGI OV 2.63 2.00 0.29 0.36
3 mm 3.08 − 6.10 − 2.03 0.76
4.5 mm 2.31 1.98 0.55 0.19
FCR OV 1.21 1.34 1.04 0.10
3 mm 1.09 1.21 1.41 0.29
4.5 mm 1.25 1.38 0.95 0.19

data, the Gompertz function and the elbow calculation estimated EFA
requirements of 1.18, 1.30 and 1.08% EPA + DHA for OV, P1 and P2,
respectively. When the NRC criterion (95% of the maximum response)
was applied to estimate EFA requirements from DGI data, much lower
values were obtained, namely 0.79, 1.04 and 0.64% EPA + DHA for OV,
P1 and P2, respectively (Fig. 3B). Using the FPL model to fit the WG
data, EFA requirements were estimated as 1.25, 1.37 and 1.19% EPA +
DHA (elbow calculation) and 1.01, 1.34 and 0.94% of dietary EPA +
DHA (NRC criterion) for OV, P1 and P2, respectively (Fig. 3C). For the
DGI data, the EFA requirements were estimated as 1.21, 1.3 and 1.09%
(elbow calculation) and 0.77, 1.05 and 0.68% of dietary EPA + DHA
(NRC criterion) for OV, P1 and P2, respectively (Fig. 3D). The fourth
parameter allowed the model to fit the growth data well in OV and P1.
However, parameter estimates for b and, or xmid had large standard
errors, due to the absence of data for the lower part of the curve. It
should be noted that for P1 the model parameters were not significant
leading to wide prediction intervals (Fig. 3D).
3.2.2. Feed conversion ratio
The split linear and quadratic models provided requirement esti­
mations that were high. This was due to the inadequacy of these models
to fit the unusual distribution of the experimental FCR data. Therefore,
the values are not discussed in detail here, but are reported in Table 5
and Fig. 4 A and B for completeness and to demonstrate the lack of fit. To
fit the Gompertz models to the FCR data it was necessary to remove data
of diet D1 (N = 15). Using the Gompertz curve to fit the FCR data, the
EFA requirements were estimated as 1.53, 1.67 and 1.47% (elbow
calculation) and 1.15, 1.92 and 1.03% EPA + DHA (NRC criterion) for
OV, P1 and P2, respectively (Fig. 4C). The removal of diet D1 data
removed error and, therefore, the model is not directly comparable to
the FPL model. Despite the unusual shape of the FCR data, the FPL model
was flexible enough to fit these data without the removal of diet D1 data.
With FCR as response, the EFA requirements were 1.55, 2.03 and 1.45%
EPA + DHA (elbow calculation) and 1.05, 1.45 and 0.99% EPA + DHA
(NRC criterion) for OV, P1 and P2, respectively (Fig. 4D). The estimated
requirement values attained for FCR were higher than for growth, and

5
S.J.S. Houston et al. Aquaculture 556 (2022) 738308

the FPL model was the only model to offer a satisfactory fit to the

size (P1, in the text) and the 4.5 mm pellet size (P2 in the text). In the Gompertz and FPL models, the EPA + DHA estimate derived from the “elbow” calculation is given. The mean squared error (MSE) and the Akaike
Requirement estimates for EPA + DHA (% of diet as fed) of juvenile gilthead seabream, Sparus aurata, from the models listed in Table 4. The models were applied to the overall data (OV, both pellet sizes), the 3 mm pellet

− 56.72
− 52.53
− 81.76
− 91.85
− 68.64
experimental data.

114.14

107.98
− 67.6
71.92
AIC
4. Discussion

0.00036
0.00021
0.00072
0.0008
0.0014
0.0018
The present study explored four different models and three perfor­

19.06

13.56
MSE

1.83
mance responses that can be used to advance estimates of EPA + DHA
requirement. The experimental design involved diets formulated with
FPL requirement
commercially relevant oils and energy densities, and strong growth
performance was observed in all dietary groups. There were two key
conclusions from this study. Firstly, it showed, within a single study that
the requirement for EPA + DHA is a function of fish mass (decreasing
1.25
1.37
1.19
1.21
1.30
1.09
1.55
2.03
1.45
with increasing weight) and could therefore be modelled with data from
more time points. Secondly, the current published requirement for EPA
+ DHA of 0.9% (dry diet) appeared too low for juvenile gilthead seab­
− 68.75
− 58.72

− 70.09
− 82.84
108.07

− 52.2

− 57.7
113.4
70.00

ream (Kalogeropoulos et al., 1992; NRC, 2011). We report the results

AIC

from different analysis methods and acknowledge that these data are
open to interpretation. However, we advance tentative recommenda­
0.00083

0.00032
0.00014
0.00073
0.0014
0.0021

tions for the two pellet sizes used in this study. For juveniles of 24–80 g,
20.44

15.22
MSE

1.84

the diet (3 mm pellet) may contain at least 1.4% EPA + DHA as a

minimum requirement but, to achieve maximum performance (highest
growth and feed conversion), the level is 2%. This level may be reduced
Gompertz requirement

in larger fish (80–250 g; 4.5 mm pellet) to 1.2% to satisfy minimum

requirement, but 1.5% to achieve maximum performance. The data
suggested that the current published guidance (~ 1%) would become
adequate at some body weight during this growth phase (P2). Further­
more, the data presented here support the reduction in the use of FM
1.22
1.34
1.18
1.18
1.30
1.08
1.53
1.67
1.47

and, or FO, provided that EFA requirements are covered in this species,
this is in line with studies using very low FM and FO levels and achieving
− 69.86
− 57.86

− 78.25

− 67.28
112.02

105.76

good growth performance, in particular in the later stages of production

70.24

− 82
AIC

(Benedito-Palos et al., 2016; Simó-Mirabet et al., 2018).

The previous requirement estimates for juvenile gilthead seabream
employed smaller fish than this study (1.2 g) and used split linear
0.00078

0.00049
0.00039
0.00089
0.0015
18.94

13.39
information criterion (AIC) are also given as an indication of how well the models fit the experimental data.

regression to analyse weight gain (g). The juvenile seabream used in the
MSE

1.86

present study attained growth rates and feed efficiency superior to the
fish than the fish used to determine the current known requirement
Split quadratic requirement

(Kalogeropoulos et al., 1992; National Research Council, 2011). As fish

performance, in particular feed efficiency increases, the requirement for
nutrients will increase to support increased growth rates (Shearer,
1995). A further study in European seabass also employed split linear
regression with DGI as the response, and reported a requirement of 0.7%
LC-PUFA (of diet dry diet) for fish of 14 g initial weight (DGI: 0.84–1.0)
(Skalli and Robin, 2004). The different approaches presented in the
2.25
1.57
1.64
2.15

3.03
6.94
2.71
1.7

present study also showed that there is no one model that suits every
–

parameter or each of the experimental periods, as suggested previously

− 67.27
− 56.22

− 80.19
− 85.29
− 70.53
109.75

101.33

(Baker, 1986; Rodehutscord and Pack, 1999).

− 57.9
70.61
AIC

The higher EFA requirements, especially for DHA, of larval fish is

relatively well understood (Izquierdo, 1996; Sargent et al., 1997). A
recent study of micronutrients and taurine in seabream showed that
0.00089

0.00044
0.00034
0.00083
0.0018
0.0015
18.67

11.67

requirements in larvae were several times higher than in fish in the grow
MSE

2.12

out phase (Izquierdo et al., 2019). The present study has shown a change
in EFA requirement between two fish (pellet) sizes and, therefore,
Split linear requirement

demonstrated the existence of a relationship between fish mass and EFA

requirement. It is likely that this relationship takes on a similar form to
digestible protein [requirement = − a ⋅ fish mass (Kg)b] (Lupatsch et al.,
2003), but to define this relationship the precise requirement for EPA +
DHA would need to be known over a range of fish masses, ideally in a
single experiment achieving optimal growth. Here, we only provide
1.21
1.47
1.13
1.20
1.46
1.09
1.97
2.50
1.62

requirement estimates for fish of around 25–80 g and 100–250 g, and so

only the negative slope between these fish weights can be estimated,
4.5 mm

4.5 mm

4.5 mm

with the value being approximately − 1.8. Alternatively, if the require­

3 mm

3 mm

3 mm
Pellet

ment is viewed per unit energy (Glencross, 2009) or lipid (Watanabe,

OV

OV

OV

1982) the decreasing requirement (% of diet) may be interpreted as a

consequence of the rising energy (DE) demand for maintenance as the
WG (g)
Table 5

Metric

fish grows (Lupatsch, 2005). Regardless of how the requirement is

FCR
DGI

numerically expressed, or even understood, its fraction in the diet (% or

6
S.J.S. Houston et al.
g kg− 1) still needs to be known to inform diet specification. The dis­
cussion below focuses on the strengths and weaknesses of the models.
The first two models applied to the data were linear and quadratic
split regression. Conceptually, these models derive the maximum
requirement because the equation describes a model approaching a
horizontal plateau, at which point the response is equivalent to the mean
of the highest performing dietary groups. For this reason, it is critical for
these techniques that the maximum response is known with at least two
dietary treatments. The exception to this is where an effect of toxicity is
observed at higher levels of nutrient supply. An example of this can be
found in black seabream (Acanthopagrus schlegelii) where high levels of
n-3 LC-PUFA led to lower growth and, in this situation, the authors used
quadratic regression without the horizontal segment (Jin et al., 2017). In
the present study, we did not observe a detrimental effect of higher EPA
+ DHA levels, so quadratic regression was not appropriate. The key
advantage with linear and quadratic split models is the provision of the
requirement by the fitting process (Glencross, 2009). Linear split
regression is the most frequently applied model in fish nutrient
requirement studies (Benedito-Palos et al., 2014; Luzzana et al., 1998;
Mai et al., 2006; Murillo-Gurrea et al., 2001; Skalli et al., 2006).
Quadratic split regression, to our knowledge, has only been applied in a
meta-analyses of fish data (Hernandez-Llamas, 2009), but there are
more examples in terrestrial animals (Hauschild et al., 2010). Interest­
ingly, the linear split model had the best fit for WG response in OV and
P2 data, and it produced EPA + DHA requirement estimates marginally
higher than the two asymptotic models (Gompertz and FPL), this being
somewhat unexpected as this method had been found to underestimate
nutrient requirements (Hernandez-Llamas, 2009; Shearer, 2000). In the
present study, the quadratic split regression gave higher estimates than
the linear split regression. Neither of these methods seemed applicable
to the FCR data due to the unusual shape of this response. As a result, the
7
Aquaculture 556 (2022) 738308
Fig. 2. The split linear (A and B) and split quadratic (C and D)
models applied to the weight gain (A and C) and daily growth
index (B and D) data against EPA + DHA % of diet (as fed, x-
axis). The raw data are marked with coloured spots, the
relevant model fit with an unbroken line, 95% prediction in­
tervals (calculated by Monte-Carlo simulation) are shaded in
colour. Colours for the different periods are as follows: the
whole trial, OV = ; the 3 mm pellet size, P1 = and the 4.5
mm pellet size, P2 = . Note, in panel D the quadratic model
would not fit the P2 data due to the high performance of D4.
Estimated requirement values are given in Table 5.
EPA + DHA requirements estimated with FCR as response variable were
considerably higher than published requirement estimates for S. aurata
(Kalogeropoulos et al., 1992) when any of the split regression models
were used. Feed efficiency/feed conversion ratio is usually given in
requirement papers, but rarely modelled as a parameter. An example
with lysine requirements of the Japanese seabass (Lateolabrax japonicus)
used split regression with feed efficiency (Mai et al., 2006).
Asymptotic functions (such as Gompertz and FPL) can often be a
strong fit to experimental data (Mercer et al., 1989) since they
adequately address the concept of diminishing returns where the in­
crease in the response slows as the plateau is reached. A limitation of the
continuous asymptotic models is that a method is required to derive a
requirement estimate, for example 0.95* asymptote (Cowey, 1992; NRC,
2011). The present results suggested that this method was highly sen­
sitive to parameter selection and might lead to erroneous requirement
estimates, at least in the context of EFA. Thus, the requirement estimates
during the OV period for both growth responses (DGI and WG) disagree
when either the FPL or Gompertz models are applied. An alternative to
the 0.95* asymptote criterion, the proposed “elbow” calculation, was
developed as a means of defining an “elbow” or optimal requirement on
a response curve. In nutrient requirement studies, it is important that
both the deficient and excess part of the nutrient response curve are
well-defined (Glencross, 2009; Shearer, 2000). It was anticipated that
the elbow calculation would be less sensitive to the selected dietary
levels of nutrient supply. Unfortunately, this is not the case and the
elbow calculation still relies on the nutrient levels determined at the
experiment’s conception. However, this calculation did seem useful in
analysing the data obtained in the present study as it gave similar
requirement estimates for both metrics of growth and both models
(albeit, marginally higher for the DGI parameter). This is due to its
insensitivity to the scale of the response. However, it is advocated that
S.J.S. Houston et al.
the elbow calculation is only applied to asymptotic functions and only to
experiments that have adequately described the deficient and excess
regions of the response. Only the elbow method has been employed
here, and we are not advocating it as a replacement method for selecting
a requirement, and there are apparent nuances with this method.
The two asymptotic models used in the present study (Gompertz and
FPL) have been used previously to fit nutrient responses in terrestrial
animals (Gahl et al., 1991; Pesti et al., 2009) but, to our knowledge, have
not been applied in fish. Other asymptotic functions (four and five-
parameter saturation kinetics models) were used in a study investi­
gating the tryptophan requirements of hybrid striped bass (Morone
chrysops x M. saxatilis) finding requirements just over 2 g kg− 1 when the
asymptote was multiplied by 0.95 (Gaylord et al., 2005). Two meta-
analyses in fish have employed a variety of non-linear models to pre­
viously published data and found that often the curves fit responses
better, but that no one model was appropriate for all data sets (Rode­
hutscord and Pack, 1999; Shearer, 2000). Therefore, we explored a
range of non-linear models and the two attaining the best fit were re­
ported here. In the present study, the Gompertz model was a good fit for
most of the growth data, especially for P1. The FPL model was also able
to fit these data, but this model carried high uncertainty because it has
parameters that are not covered by the data (b and xmid). To apply the
Gompertz model to the FCR data it was necessary to disregard data for
diet D1 and, therefore, it does not capture reality well, which created
problems comparing fits as error associated with diet D1 was discarded.
The only model that could describe the FCR response to the diets was the
FPL model and, as such, requirements for FCR differed between the two
asymptotic models and it is difficult to advance an estimate, although it
can be said that the requirement to achieve the best FCR was higher than
for the growth metrics.
It is evident from these empirical data that the current published
8
Aquaculture 556 (2022) 738308
Fig. 3. The non-linear Gompertz model (A and B) and four
parameter logistic model (FPL in the text, C and D) applied to
the weight gain (A and C) and daily growth index (B and D)
data against EPA + DHA % of diet (as fed, x-axis). The raw data
are marked with coloured spots, the relevant model fit with an
unbroken line, requirements with black symbols and the 95%
prediction intervals (calculated with 2nd order Taylor expan­
sion) are shaded in colour. Symbols for the requirement esti­
mates are as follows: Published = ■, National Research
Council = ▴ (NRC criterion in text) and Elbow = ●. Colours for
the different periods are as follows: the whole trial, OV = ;
the 3 mm pellet size, P1 = and the 4.5 mm pellet size, P2 =
. Note in panel D in P1, the error was not plotted for clarity
because it was very wide, an indication that the curve for this
data was over-parameterised. Estimated requirement values
are given in Table 5.
requirement of 0.9 (dry weight) / 1% EPA + DHA is too low for juveniles
of this species, especially when considering fish of 25–80 g. The present
study has demonstrated that no model is a fit for each situation and the
NRC definition of a requirement at 95% of a response maximum may not
be entirely appropriate for EPA + DHA requirements. One reason for this
is that the models suggest that n-3 LC-PUFA (i.e. EPA and DHA) are not
completely essential to growth (all models have positive intercepts,
implying that there is no maintenance demand and no weight loss), at
least over the timescale and EFA levels used for this study. Interestingly,
results modelling the turnover of individual fatty acids in Asian seabass
(L. calcarifer) suggested zero maintenance requirement for EPA and DHA
(Salini et al., 2016). The elbow-calculation was applied to determine a
point on an asymptotic response curve that was less sensitive to the
design of the experiment although, unfortunately, it is still not insensi­
tive to experimental design. The calculation, however, does bisect the
curve well and the data demonstrated it returned values that agreed with
each other across models and metrics. However, its application was not
appropriate unless the maximum response and deficient responses were
well defined and we only advocate its use for asymptotic models. The
elbow calculation defines a point on the curve that further increases in
dietary EPA + DHA levels lead to little gain in the response and,
therefore, denotes an optimum requirement (Fig. 1).
In summary, the key conclusions of this work are that 1) S. aurata
juveniles of 25 g – 80 g (3 mm pellet) have a higher requirement for EPA
+ DHA of at least 1.4% of diet for optimal/maximum growth, but
perhaps as high as 2% if FCR is considered, 2) the requirements for EPA
+ DHA are a function of fish mass, the implication of this being that
requirements could be modelled but the trial would need a larger
number of sampling points than the two reported here and, 3) the exact
definition of EPA + DHA requirement is a question of context; mainte­
nance, optimum production, maximum performance, health, well-being
S.J.S. Houston et al.
or product quality could all ultimately reflect different end-points and,
therefore, give different estimated requirements.
Authorship
VK and JT designed and executed the nutritional trial and all authors
contributed to planning the analyses. SJSH, VK, and JT carried out the
sampling. OM and DRT supervised the lead author and BG provided
technical expertise during manuscript preparation. SJSH analysed the
data and prepared the draft manuscript, which was then shared between
all authors who made amendments, contributions and
recommendations.
CRediT authorship contribution statement
Sam J.S. Houston: Investigation, Methodology, Formal analysis,
Writing – original draft, Visualization. Vasileios Karalazos: Concep­
tualization, Investigation, Methodology, Supervision, Writing – review
& editing, Funding acquisition. John Tinsley: Conceptualization,
Investigation, Methodology, Supervision, Writing – review & editing,
Funding acquisition. Douglas R. Tocher: Conceptualization, Investi­
gation, Supervision, Writing – review & editing, Funding acquisition.
Brett D. Glencross: Writing – review & editing. Óscar Monroig:
Conceptualization, Investigation, Supervision, Writing – review & edit­
ing, Funding acquisition.
Declaration of Competing Interest
None.
9
Aquaculture 556 (2022) 738308
Fig. 4. The linear (A) and quadratic (B) split regression models
applied to the FCR data against EPA + DHA % of diet (as fed, x-
axis), the 95% prediction intervals (shaded colour) were
calculated by Monte-Carlo simulation. The non-linear Gom­
pertz (C) and four parameter logistic (FPL in the text, D)
applied the FCR data against EPA + DHA % of diet (as fed, x-
axis), the 95% prediction intervals (shaded colour) were
calculated by second-order Taylor expansion. The raw data are
marked with coloured spots, the relevant model fit with an
unbroken line and requirements with black symbols. Symbols
for the requirement estimates are as follows: Published = ■,
National Research Council = ▴ (NRC criterion in text) and
Elbow = ●. Colours for the different periods are as follows: the
whole trial, OV = ; the 3 mm pellet size, P1 = and the 4.5
mm pellet size, P2 = . Note in panel B, the error for P1 was
not plotted, but clearly it is a poor fit to the data. Estimated
requirement values are given in Table 5.
Acknowledgements
This work was co-funded by BioMar AS and the Marine Alliance for
Science and Technology Scotland (MASTS). BioMar provided the
experimental feeds, trial facilities and fish, and covered travel expenses.
The authorsexpress thanks to the technical staff at the BioMar Feed Trial
Unit, Hirtshals, Denmark for expert care of the fish during this trial.
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are very valuable for some kinds of medical work. The application of
these currents is quite painless, and but for the strange-looking
apparatus the patient probably would not know that anything unusual
was taking place. To some extent the effect maybe said to be not
unlike that of a powerful tonic. Insomnia and other troubles due to
disordered nerves are quickly relieved, and even such obstinate
complaints as neuritis and crippling rheumatism have been cured.
The treatment is also of great value in certain forms of heart trouble.
By increasing the strength of the high frequency currents the tissues
actually may be destroyed, and this power is utilized for
exterminating malignant growths, such as lupus or cancer.
The heat produced by a current of electricity is made use of in
cauterizing. The burner is a loop of platinum wire, shaped according
to the purpose for which it is intended, and it is used at a dull red
heat. Very tiny electric incandescent lamps, fitted in long holders of
special shape, are largely used for examining the throat and the
various cavities of the body.
In the Finsen light treatment electric light is used for a very
different purpose. The spectrum of white light consists of the colours
red, orange, yellow, green, blue, indigo, and violet. Just beyond the
violet end of the spectrum are the ultra-violet rays. Ultra-violet light
consists of waves of light which are so short as to be quite invisible
to the eye, and Dr. N. R. Finsen, a Danish physician, made the
discovery that this light is capable of destroying bacterial germs. In
the application of ultra-violet rays to medical work, artificial light is
used in preference to sunlight; for though the latter contains ultra-
violet light, a great deal of it is absorbed in passing through the
atmosphere. Besides this, the sun sends out an immense amount of
radiant heat, and this has to be filtered out before the light can be
used. The usual source of light is the electric arc, and the arc is
much richer in ultra-violet rays if it is formed between electrodes of
iron, instead of the usual carbon rods. The light, which, in addition to
the ultra-violet rays, includes the blue, indigo, and violet parts of the
spectrum, is passed along a tube something like that of a telescope,
and is focused by means of a double lens, consisting of two separate
plates of quartz. Glass cannot be used for the lens, because it is
opaque to the extreme ultra-violet rays. A constant stream of water is
passed between the two plates forming the lens, and this filters out
the heat rays, which are not wanted. In some forms of Finsen lamp
an electric spark is used as the source of light, in place of the arc.
The most important application of the Finsen light is in the cure
of the terribly disfiguring disease called lupus. This is a form of
tuberculosis of the skin, and it is produced by the same deadly
microbe which, when it attacks the lungs, causes consumption. In all
but extreme cases the Finsen light effects a remarkable cure. A
number of applications are necessary, each of half an hour or more;
and after a time the disease begins to disappear, leaving soft, normal
skin. The exact action of the light rays is a disputed point. Finsen
himself believed that the ultra-violet rays attacked and exterminated
the microbe, but a later theory is that the rays stimulate the tissues to
such an extent that they are enabled to cure themselves. As early as
the year 1899 Finsen had employed his light treatment in 350 cases
of lupus, and out of this number only five cases were unsuccessful.
The ultra-violet rays are said to have a very beneficial effect
upon the teeth. Experiments carried out in Paris, using a mercury
vapour lamp as the source of light, show that discoloured teeth are
whitened and given a pearly lustre by these rays, at the same time
being sterilized so that they do not easily decay. The Röntgen rays
are used for the treatment of lupus, and more particularly for deeper
growths, such as tumours and cancers, for which the Finsen rays are
useless, owing to their lack of penetrating power. The action of these
two kinds of rays appears to be similar, but the X-rays are much the
more active of the two.
Electricity is often applied to the body through water, in the form
of the hydro-electric bath, and such baths are used in the treatment
of different kinds of paralysis. Electric currents are used too for
conveying drugs into the tissues of the body. This is done when it is
desired to concentrate the drug at some particular point, and it has
been found that chemicals can be forced into the tissues for a
considerable distance.
 Dr. Nagelschmidt, a great authority on medical electricity, has
suggested the use of electricity for weight reducing. In the ordinary
way superfluous flesh is got rid of by a starvation diet coupled with
exercise, but in many cases excessively stout people are troubled
with heart disorders and asthma, so that it is almost impossible for
them to undergo the necessary muscular exertion. By the application
of electric currents, however, the beneficial effects of the gentle
exercise may be produced without any exertion on the part of the
patient, and an hour’s treatment is said to result in a decrease in
weight of from 200 to 800 grammes, or roughly 7 to 27 ounces.
 CHAPTER XXVI
OZONE

The great difference between the atmospheric conditions before and

after a thunderstorm must have been noticed by everybody. Before
the storm the air feels lifeless. It does not satisfy us as we draw it
into our lungs, and however deeply we breathe, we feel that
something is lacking. After the storm the air is delightful to inhale,
and it refreshes us with every breath. This remarkable transformation
is brought about to a very large extent by ozone produced by the
lightning discharges.
As far back as 1785 it was noticed that oxygen became changed
in some way when an electric spark was passed through it, and that
it acquired a peculiar odour. No particular attention was paid to the
matter however until about 1840, when Schönbein, a famous
German chemist, and the discoverer of gun-cotton and collodion,
became interested in it. He gave this strange smelling substance the
name of “ozone,” and he published the results of his experiments
with it in a treatise entitled, “On the Generation of Ozone.”
Schönbein showed that ozone could be produced by various
methods, chemical as well as electrical. For instance, if a piece of
phosphorus is suspended in a jar of air containing also a little water,
in such a manner that it is partly in the water and partly out of it, the
air acquires the characteristic smell of ozone, and it is found to have
gained increased chemical energy, so that it is a more powerful
oxidizing agent. For a long time the exact chemical nature of ozone
could not be determined, mainly because it was impossible to obtain
the substance in quantities sufficiently large for extensive
experimental research, but also on account of its extremely energetic
properties, which made it very troublesome to examine. These
difficulties were so great that investigators were in doubt as to
whether ozone was an element or a compound of two or more
elements; but finally it was proved that it was simply oxygen in a
condensed or concentrated state.
Apparently ozone is formed by the contraction of oxygen, so that
from three volumes of oxygen two volumes of ozone are produced.
In other words, ozone has one and a half times the density of
oxygen. Ozone has far greater oxidizing power than oxygen itself; in
fact it is probably the most powerful of all oxidizing agents, and
herein lies its great value. It acts as nature’s disinfectant or sterilizer,
and plays a very important part in keeping the air pure, by destroying
injurious organic matter. Bacteria apparently have a most decided
objection to dying; at any rate they take an extraordinary amount of
killing. Ozone is more than a match for them however, and under its
influence they have a short life and probably not a merry one.
Ozone exists naturally in the atmosphere in the open country,
and more especially at the seaside. It is produced by lightning
discharges, by silent electrical discharges in the atmosphere, by the
evaporation of water, particularly salt water, by the action of sunlight,
and also by the action of certain vegetable products upon the air.
The quantity of ozone in the air is always small, and even pure
country or sea air contains only one volume of ozone in about
700,000 volumes of air. No ozone can be detected in the air of large
towns, or over unhealthy swamps or marshes. The exhilarating
effects of country and sea air, and the depressing effects of town air,
are due to a very large extent to the presence or absence of ozone.
A great proportion of our common ailments are caused directly
or indirectly by a sort of slow poisoning, produced by the impure air
in which we live and work. It is popularly supposed that the tainting
of the air of rooms in which large numbers of people are crowded
together is due to an excessive amount of carbonic acid gas. This is
a mistake, for besides being tasteless and odourless, carbonic acid
gas is practically harmless, except in quantities far greater than ever
exist even in the worst ventilated rooms. The real source of the
tainted air is the great amount of animal matter thrown off as waste
products from the skin and lungs, and this tainting is further
intensified by the absence of motion in the air. Even in an over-
crowded room the conditions are made much more bearable if the
air is kept in motion, and in a close room ladies obtain relief by the
use of their fans. What we require, therefore, in order to maintain an
agreeable atmosphere under all conditions, is some means of
keeping the air in gentle motion, and at the same time destroying as
much as possible of the animal matter contained in it. Perhaps the
most interesting and at the same time the most scientific method of
doing this is by ozone ventilation.
In the well-known “Ozonair” system of ventilation, ozone is
generated by high-tension current. Low-tension current is taken from
the public mains or from accumulators, and raised to a very high
voltage by passing it through a step-up transformer. The secondary
terminals of the transformer are connected to a special form of
condenser, consisting of layers of fine metal gauze separated by an
insulating substance called “micanite.” The high tension between the
gauze layers produces a silent electrical discharge or glow. A small
fan worked by an electric motor draws the air over the condenser
plates, and so a certain proportion of the oxygen is ozonized, and is
driven out of the other side of the apparatus into the room. The
amount of ozone generated and the amount of air drawn over the
condenser are regulated carefully, so that the ozonized air contains
rather less than one volume of ozone in one million volumes of air,
experiment having shown that this is the most suitable strength for
breathing. Ozone diluted to this degree has a slight odour which is
very refreshing, and besides diminishing the number of organic
germs in the air, it neutralizes unpleasant smells, such as arise from
cooking or stale tobacco smoke. Ozone ventilation is now employed
successfully in many hotels, steamships, theatres and other places
of entertainment, municipal and public buildings, and factories.
By permission of] [Ozonair, Ltd.

Fig. 42.—Diagram of Ozonizing Plant, Central London Tube Electric Railway.

One of the most interesting examples of ozone ventilation is that

of the Central London tube electric railway. The installation consists
of a separate ozonizing plant at every station, except Shepherd’s
Bush, which is close to the open end of the tunnel. Fig. 42 is a
diagram of the general arrangement of one of these plants, and it
shows how the air is purified, ozonized, and sent into the tunnel. The
generating plant is seen at the top left-hand corner of the figure. Air
is drawn in as shown by the arrows, and by passing through the filter
screen F it is freed from dirt and smuts, and from most of the
injurious gases which always are present in town air. The filter
screen is kept moist by a continual flow of water from jets above it,
the waste water falling into the trough W. The ozone generator is
shown at O. Continuous current at about 500 volts, from the power
station, is passed through a rotary converter, which turns it into
alternating current at 380 volts. This current goes to the transformer
T, from which it emerges at a pressure of 5000 volts, and is supplied
to the ozone generator. From the generator the strongly ozonized air
is taken by way of the ozone pipe P, to the mixing chamber of the
large ventilating fan M, where it is mixed with the main air current
and then blown down the main air trunk. From this trunk it is
distributed to various conduits, and delivered at the air outlets
marked A. Altogether the various plants pump more than eighty
million cubic feet of ozonized air into the tunnels every working day.
In many industries pure air is very essential, especially during
certain processes. This is the case in brewing, in cold storage, and in
the manufacture and canning of food products; and in these
industries ozone is employed as an air purifier, with excellent results.
Other industries cannot be carried on without the production of very
unpleasant fumes and smells, which are a nuisance to the workers
and often also to the people living round about; and here again
ozone is used to destroy and remove the offending odours. It is
employed also in the purification of sewage and polluted water; in
bleaching delicate fabrics; in drying and seasoning timber; in
maturing tobacco, wines and spirits, and in many other processes
too numerous to mention.
 CHAPTER XXVII
ELECTRIC IGNITION

The petrol motor, which to-day is busily engaged all over the world in
driving thousands upon thousands of self-propelled vehicles or
automobiles, belongs to the important class of internal-combustion
engines. Combustion means the operation of burning, and an
internal-combustion engine is one in which the motive power is
produced by the combustion of a highly explosive mixture of gases.
In the ordinary petrol motor this mixture consists of petrol and air,
and it is made by means of a device called a “carburetter.” By
suction, a quantity of petrol is forced through a jet with a very fine
nozzle, so that it is reduced to an extremely fine spray. A certain
proportion of air is allowed to enter, and the mixture passes into the
cylinder. Here it is compressed by the rising piston so that it
becomes more and more heated, and at the right point it is ignited.
Combustion takes place with such rapidity that it takes the form of an
explosion, and the energy produced in this way drives forward the
piston, which turns the crank-shaft and so communicates motion to
the driving-wheels.
The part played by electricity in this process is confined to the
ignition of the compressed charge of petrol and air. This may be
done in two ways; by means of an accumulator and a small induction
coil, or by means of a dynamo driven by the engine. At one time the
first method was employed exclusively, but to-day it is used as a rule
only for starting the car engine, the second or magneto method
being used when the engine has started up.
In accumulator ignition the low-tension current from the
accumulator passes through an induction coil, and is thus
transformed to high-tension current. This current goes through a
sparking plug, which is fixed in the head of the cylinder. The sparking
plug contains two metal points separated by a tiny air gap of from
about 1/30 to 1/50 inch. This gap provides the only possible path for
the high-tension current, so that the latter leaps across it in the form
of a spark. The spark is arranged to take place when the piston is at
the top of its stroke, that is, when the explosive mixture is at its
maximum compression, and the heat of the spark ignites the
mixture, the resulting explosion forcing down the piston with great
power. In practice it is found better as a rule to cause the spark to
pass very slightly before the piston reaches the extreme limit of its
stroke. The reason of this is that the process of igniting and
exploding the charge occupies an appreciable, though of course
exceedingly small amount of time. Immediately on reaching the top
of its stroke the piston begins to descend again, and if the spark and
the top of the stroke coincide in time the explosion does not take
place until the piston has moved some little distance down the
cylinder, and so a certain amount of power is lost. By having the
spark a little in advance of the piston, the explosion occurs at the
instant when the piston begins to return, and so the full force of the
explosion is utilized.
In magneto ignition the current is supplied by a small dynamo.
This generates alternating current, and it is driven by the car engine.
The current is at first at low pressure, and it has to be transformed to
high-tension current in order to produce the spark. There are two
methods of effecting this transformation. One is by turning the
armature of the dynamo into a sort of induction coil, by giving it two
separate windings, primary and secondary; so that the dynamo
delivers high-tension current directly. The other method is to send
the low-tension current through one or more transformer coils, just
as in accumulator ignition. Accumulators can give current only for a
certain limited period, and they are liable consequently to run down
at inconvenient times and places. They also have the defect of
undergoing a slight leakage of current even when they are not in
use. Magneto ignition has neither of these drawbacks, and on
account of its superior reliability it has come into universal use.
 In the working of quarries and mines of various kinds, and also in
large engineering undertakings, blasting plays a prominent part.
Under all conditions blasting is a more or less dangerous business,
and it has been the cause of very many serious accidents to the men
engaged in carrying it out. Many of these accidents are due to the
carelessness resulting from long familiarity with the work, but apart
from this the danger lies principally in uncertainty in exploding the
charge. Sometimes the explosion occurs sooner than expected, so
that the men have not time to get away to a safe distance. Still more
deadly is the delayed explosion. After making the necessary
arrangements the men retire out of danger, and await the explosion.
This does not take place at the expected time, and after waiting a
little longer the men conclude that the ignition has failed, and return
to put matters right. Then the explosion takes place, and the men are
killed instantly or at least seriously injured. Although it is impossible
to avoid altogether dangers of this nature, the risk can be reduced to
the minimum by igniting the explosives by electricity.
Electrical shot firing may be carried out in different ways,
according to circumstances. The current is supplied either by a
dynamo or by a battery, and the firing is controlled from a
switchboard placed at a safe distance from the point at which the
charge is to be exploded, the connexions being made by long
insulated wires. The actual ignition is effected by a hot spark, as in
automobile ignition, or by an electric detonator or fuse. Explosives
such as dynamite cannot be fired by simple ignition, but require to be
detonated. This is effected by a detonator consisting of a small cup-
shaped tube, made of ebonite or other similar material. The wires
conveying the current project into this tube, and are connected by a
short piece of very fine wire having a high resistance. Round this
wire is packed a small quantity of gun-cotton, and beyond, in a sort
of continuation of the tube, is placed an extremely explosive
substance called “fulminate of mercury,” the whole arrangement
being surrounded by the dynamite to be fired. When all is ready the
man at the switchboard manipulates a switch, and the current
passes to the detonator and forces its way through the resistance of
the thin connecting wire. This wire becomes sufficiently hot to ignite
the gun-cotton, and so explode the fulminate of mercury. The
explosion is so violent that the dynamite charge is detonated, and
the required blasting carried out. Gunpowder and similar explosives
do not need to be detonated, and so a simple fuse is used. Electric
fuses are much the same as detonators, except that the tube
contains gunpowder instead of fulminate of mercury, this powder
being ignited through an electrically heated wire in the same way.
These electrical methods do away with the uncertainty of the slow-
burning fuses formerly employed, which never could be relied upon
with confidence.
Enormous quantities of explosives are now used in blasting on a
large scale, where many tons of hard rock have to be removed. One
of the most striking blasting feats was the blowing up of Flood Island,
better known as Hell Gate. This was a rocky islet, about 9 acres in
extent, situated in the East River, New York. It was a continual
menace to shipping, and after many fine vessels had been wrecked
upon it the authorities decided that it should be removed. The rock
was bored and drilled in all directions, the work taking more than a
year to complete; and over 126 tons of explosives were filled into the
borings. The exploding was carried out by electricity, and the mighty
force generated shattered nearly 300,000 cubic yards of solid rock.
 CHAPTER XXVIII
ELECTRO-CULTURE

About thirty years ago a Swedish scientist, Professor Lemström,

travelled extensively in the Polar regions, and he was greatly struck
by the development of the Polar vegetation. In spite of the lack of
good soil, heat, and light, he observed that this vegetation came to
maturity quicker than that of regions having much more favourable
climates, and that the colours of the flowers were remarkably fresh
and clear, and their perfumes exceptionally strong. This was a
surprising state of things, and Lemström naturally sought a clue to
the mystery. He knew that peculiar electrical conditions prevailed in
these high latitudes, as was shown by the wonderful displays of the
Aurora Borealis, and he came to the conclusion that the
development of the vegetation was due to small currents of
electricity continually passing backwards and forwards between the
atmosphere and the Earth. On his return to civilization Lemström at
once began a series of experiments to determine the effect of
electricity upon the growth of plants, and he succeeded in proving
beyond all doubt that plants grown under electrical influence
flourished more abundantly than those grown in the ordinary way.
Lemström’s experiments have been continued by other investigators,
and striking and conclusive results have been obtained.
The air surrounding the Earth is always charged to some extent
with electricity, which in fine weather is usually positive, but changes
to negative on the approach of wet weather. This electricity is always
leaking away to the earth more or less rapidly, and on its way it
passes through the tissues of the vegetation. An exceedingly slow
but constant discharge therefore is probably taking place in the
tissues of all plants. Experiments appear to indicate that the upper
part of a growing plant is negative, and the lower part positive, and at
any rate it is certain that the leaves of a plant give off negative
electricity. In dull weather this discharge is at its minimum, but under
the influence of bright sunshine it goes on with full vigour. It is not
known exactly how this discharge affects the plant, but apparently it
assists its development in some way, and there is no doubt that
when the discharge is at its maximum the flow of sap is most
vigorous. Possibly the electricity helps the plant to assimilate its
food, by making this more readily soluble.
This being so, a plant requires a regular daily supply of
uninterrupted sunshine in order to arrive at its highest possible state
of maturity. In our notoriously variable climate there are many days
with only short intermittent periods of bright sunshine, and many
other days without any sunshine at all. Now if, on these dull days, we
can perform at least a part of the work of the sunshine, and
strengthen to some extent the minute currents passing through the
tissues of a plant, the development of this plant should be
accelerated, and this is found to be the case. Under electrical
influence plants not only arrive at maturity quicker, but also in most
cases their yield is larger and of finer quality.
Lemström used a large influence machine as the source of
electricity in his experiments in electro-culture. Such machines are
very suitable for experimental work on a small scale, and much
valuable work has been done with them by Professor Priestly and
others; but they have the great drawback of being uncertain in
working. They are quite satisfactory so long as the atmosphere
remains dry, but in damp weather they are often very erratic, and
may require hours of patient labour to coax them to start. For this
reason an induction coil is more suitable for continuous work on an
extensive scale.
The most satisfactory apparatus for electro-culture is that used in
the Lodge-Newman method, designed by Sir Oliver Lodge and his
son, working in conjunction with Mr. Newman. This consists of a
large induction coil supplied with current from a dynamo driven by a
small engine, or from the public mains if available. This coil is fitted
with a spark gap, and the high-tension current goes through four or
five vacuum valve globes, the invention of Sir Oliver Lodge, which
permit the current to pass through them in one direction only. This is
necessary because, as we saw in Chapter VIII., two opposite
currents are induced in the secondary winding of the coil, one at the
make and the other at the break of the primary circuit. Although the
condenser fitted in the base of the coil suppresses to a great extent
the current induced on making the circuit, still the current from the
coil is not quite uni-directional, but it is made so by the vacuum
rectifying valves. These are arranged to pass only the positive
current, and this current is led to overhead wires out in the field to be
electrified. Lemström used wires at a height of 18 inches from the
ground, but these were very much in the way, and in the Lodge-
Newman system the main wires are carried on large porcelain
insulators fixed at the top of poles at a height of about 15 feet. This
arrangement allows carting and all other agricultural operations to be
carried on as usual. The poles are set round the field, about one to
the acre, and from these main wires finer ones are carried across the
field. These wires are placed about 30 feet apart, so that the whole
field is covered by a network of wires. The electricity supplied to the
wires is at a pressure of about 100,000 volts, and this is constantly
being discharged into the air above the plants. It then passes
through the plants, and so reaches the earth. This system may be
applied also to plants growing in greenhouses, but owing to the
confined space, and to the amount of metal about, in the shape of
hot-water pipes and wires for supporting plants such as vines and
cucumbers, it is difficult to make satisfactory arrangements to
produce the discharge.
The results obtained with this apparatus at Evesham, in
Gloucestershire, by Mr. Newman, have been most striking. With
wheat, increases of from 20 per cent. to nearly 40 per cent. have
been obtained, and the electrified wheat is of better quality than
unelectrified wheat grown at the same place, and, apart from
electrification, under exactly the same conditions. In some instances
the electrified wheat was as much as 8 inches higher than the
unelectrified wheat. Mr. Newman believes that by electrification land
yielding normally from 30 to 40 bushels of wheat per acre can be
made to yield 50 or even 60 bushels per acre. With cucumbers
under glass increases of 17 per cent. have been obtained, and in the
case of strawberries, increases of 36 per cent. with old plants, and
80 per cent. with one-year-old plants. In almost every case
electrification has produced a marked increase in the crop, and in
the few cases where there has been a decrease the crops were
ready earlier than the normal. For instance, in one experiment with
broad beans a decrease of 15 per cent. resulted, but the beans were
ready for picking five days earlier. In another case a decrease of 11½
per cent. occurred with strawberries, but the fruit was ready for
picking some days before the unelectrified fruit, and also was much
sweeter. In some of the experiments resulting in a decrease in the
yield it is probable that the electrification was overdone, so that the
plants were over-stimulated. It seems likely that the best results will
be obtained only by adjusting the intensity and the duration of the
electrification in accordance with the atmospheric conditions, and
also with the nature of the crop, for there is no doubt that plants vary
considerably in their electrical requirements. A great deal more
experiment is required however to enable this to be done with
anything like certainty.
Unlike the farmer, the market gardener has to produce one crop
after another throughout the year. To make up for the absence of
sufficient sunshine he has to resort to “forcing” in many cases, but
unfortunately this process, besides being costly, generally results in
the production of a crop of inferior quality. Evidently the work of the
market gardener would be greatly facilitated by some artificial
substitute for sunshine, to keep his plants growing properly in dull
weather. In 1880, Sir William Siemens, knowing that the composition
of the light of the electric arc was closely similar to that of sunlight,
commenced experiments with an arc lamp in a large greenhouse.
His idea was to add to the effects of the solar light by using the arc
lamp throughout the night. His first efforts were unsuccessful, and he
discovered that this was due to the use of the naked light, which
apparently contained rays too powerful for the plants. He then
passed the light through glass, which filtered out the more powerful
rays, and this arrangement was most successful, the plants
responding readily to the artificial light. More scientifically planned
experiments were carried out at the London Royal Botanic Gardens
in 1907, by Mr. B. H. Thwaite, and these showed that by using the
arc lamp for about five hours every night, a great difference between
the treated plants and other similar plants grown normally could be
produced in less than a month. Other experiments made in the
United States with the arc lamp, and also with ordinary electric
incandescent lamps, gave similar results, and it was noticed that the
improvement was specially marked with cress, lettuce, spinach, and
other plants of this nature.
In 1910, Miss E. C. Dudgeon, of Dumfries, commenced a series
of experiments with the Cooper-Hewitt mercury vapour lamp. Two
greenhouses were employed, one of which was fitted with this lamp.
Seeds of various plants were sown in small pots, one pot of each
kind being placed in each house. The temperature and other
conditions were kept as nearly alike as possible in both houses, and
in the experimental house the lamp was kept going for about five
hours every night. In every case the seeds in the experimental house
germinated several days before those in the other house, and the
resulting plants were healthy and robust. Later experiments carried
out by Miss Dudgeon with plants were equally successful.
From these experiments it appears that the electric arc, and still
more the mercury vapour lamp, are likely to prove of great value to
the market gardener. As compared with the arc lamp, the mercury
vapour lamp has the great advantage of requiring scarcely any
attention, and also it uses less current. Unlike the products of
ordinary forcing by heat, the plants grown under the influence of the
mercury vapour light are quite sturdy, so that they can be planted out
with scarcely any “hardening off.” The crop yields too are larger, and
of better quality. The wonderful effects produced by the Cooper-
Hewitt lamp are certainly not due to heat, for this lamp emits few
heat rays. The results may be due partly to longer hours worked by
the plants, but this does not explain the greater accumulation of
chlorophyll and stronger development of fibre.
Most of us are familiar with the yarn about the poultry keeper
who fitted all his nests with trap-doors, so that when a hen laid an
egg, the trap-door opened under the weight and allowed the egg to
fall through into a box lined with hay. The hen then looked round,
and finding no egg, at once set to work to lay another. This in turn
dropped, another egg was laid, and so on. It is slightly doubtful
whether the modern hen could be swindled in this bare-faced
manner, but it is certain that she can be deluded into working
overtime. The scheme is absurdly simple. Electric lamps are fitted in
the fowl-house, and at sunset the light is switched on. The
unsuspecting hens, who are just thinking about retiring for the night,
come to the conclusion that the day is not yet over, and so they
continue to lay. This is not a yarn, but solid fact, and the increase in
the egg yield obtained in this way by different poultry keepers ranges
from 10 per cent. upwards. Indeed, one poultry expert claims to have
obtained an increase of about 40 per cent.
The ease with which a uniform temperature can be maintained
by electric heating has been utilized in incubator hatching of
chickens. By means of a specially designed electric radiator the
incubator is kept at the right temperature throughout the hatching
period. When the chickens emerge from the eggs they are
transferred to another contrivance called a “brooder,” which also is
electrically heated, the heat being decreased gradually day by day
until the chicks are sturdy enough to do without it. Even at this stage
however the chickens do not always escape from the clutches of
electricity. Some rearers have adopted the electric light swindle for
the youngsters, switching on the light after the chickens have had a
fair amount of slumber, so that they start feeding again. In this way
the chickens are persuaded to consume more food in the twenty-four
hours, and the resulting gain in weight is said to be considerable.
More interesting than this scheme is the method of rearing chickens
under the influence of an electric discharge from wires supplied with
high-tension current. Comparative tests show that electrified
chickens have a smaller mortality and a much greater rate of growth
than chickens brought up in the ordinary way. It even is said that the
electrified chickens have more kindly dispositions than their
unelectrified relatives!
Possibly the high-tension discharge may turn out to be as
beneficial to animals as it has been proved to be for plants, but so far
there is little reliable evidence on this point, owing to lack of
experimenters. A test carried out in the United States with a flock of
sheep is worth mention. The flock was divided into two parts, one-
half being placed in a field under ordinary conditions, and the other
in a field having a system of overhead discharge wires, similar to
those used in the Lodge-Newman system. The final result was that
the electrified sheep produced more than twice as many lambs as
the unelectrified sheep, and also a much greater weight of wool. If
further experiments confirm this result, the British farmer will do well
to consider the advisability of electrifying his live-stock.
 CHAPTER XXIX
SOME RECENT APPLICATIONS OF
ELECTRICITY—AN ELECTRIC PIPE LOCATOR

One of the great advantages of living in a town is the abundant

supply of gas and water. These necessary substances are conveyed
to us along underground pipes, and a large town has miles upon
miles of such pipes, extending in all directions and forming a most
complex network. Gas and water companies keep a record of these
pipes, with the object of finding any pipe quickly when the necessity
arises; but in spite of such records pipes are often lost, especially
where the whole face of the neighbourhood has changed since the
pipes were laid. The finding of a lost pipe by digging is a very
troublesome process, and even when the pipe is known to be close
at hand, it is quite surprising how many attempts are frequently
necessary before it can be located, and its course traced. As may be
imagined, this is an expensive business, and often it has been found
cheaper to lay a new length of pipe than to find the old one. There is
now an electrical method by which pipe locating is made
comparatively simple, and unless it is very exceptionally deep down,
a pipe never need be abandoned on account of difficulty in tracing it.
The mechanism of an electric pipe locator is not at all
complicated, consisting only of an induction coil with battery, and a
telephone receiver connected to a coil of a large number of turns of
thin copper wire. If a certain section of a pipe is lost, and has to be
located, operations are commenced from some fitting known to be
connected with it, and from some other fitting which may or may not
be connected with the pipe, but which is believed to be so
connected. The induction coil is set working, and its secondary
terminals are connected one to each of these fittings. If the second