J. Env. Bio-Sci., 2015: Vol.

29 (2):283-288
(283) ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

COLOUR ENHANCEMENT OF SWORD TAIL [XIPHOPHORUS HELLERI (HECKEL,

1848)] BY USING FEED FORMULATED WITH SELECTED ORNAMENTAL FLOWERS
Golandaj A.1, Swain S.1 and Mogalekar H. S.2
1
School of Aquaculture and Biotechnology, Faculty of Fisheries, Kerala University of Fisheries and Ocean Studies,
Panangad, Kochi - 682506, Kerala, India.
2
Department of Fisheries Biology and Resource Management, Fisheries College & Research Institute, Tamil Nadu Fisheries
University, Thoothukudi-628 008, Tamil Nadu, India.
[Corresponding author E-mail2: mogalekar.hs10@gmail.com]

Received: 19-08-2015 Accepted: 19-10-2015

The present study (90 days) was conducted to evaluate the colour enhancement of Xiphophorus helleri by using feed formulated
with selected ornamental flowers. Completely randomized design was employed for the study. Dried petal meal of the three
chosen ornamental sources was incorporated in the test diets at 5% each. The test diets were designated as T1, T2 and T3
respectively. A basal feed without carotenoid served as the control (T0). Ten larvae of sword tail were stocked in each tank. Each
diet was fed to fish in a set of three tanks each. Growth response as well as water quality parameters were monitored at fortnightly
intervals. The growth response was the highest in marigold supplemented diets followed by ixora and hibiscus supplemented
diets. The colour enhancement was assessed by colorimetry as well as by employing computer assisted approach, employing
Adobe Photoshop® Version 7. On termination of the study, the growth data was statistically analysed using two way analysis of
variance. Fish fed all the three test diets showed significantly different growth response from the control fish. Significant enhancement
in colour development was observed in fish fed all three test diets. The intensity of colour development was highest in T1 treatment
followed by T2 and T3. Fish fed the control diet showed the least colour intensity. The three flower meals assessed are cheap,
easily available and involve only marginal additional cost. This approach is easily adoptable at the farmer level and moreover, does
not involve any adverse effects. Further studies are required on the optimum level of inclusion of the carotenoid sources in the
diets.

Ornamental fish keeping is becoming popular as a relaxing de novo3. A variety of carotenoid both synthetic and naturally
and stress relieving hobby in the world. Ornamental fishes are occurring products from natural sources contain mixture of
gaining an important component of aquaculture because of several carotenoid like α -carotenoid, β -carotenoid,
their aesthetic value and also due to their immense commercial zeaxanthin, lutein and cryptoxanthin; whereas synthetic
value in the export trade world over. The aquarium fish trade processes provides only specific carotenoid like β -carotene.
with a turnover of US$ 5 billion in 1998 and an annual growth
In this context, the present study was conducted to evaluate
rate of 10 percent per year has increased today to US$ 7
the color enhancement potential of three indigenous flower
billion1. The bright external coloration of fish in particular has
pigment sources namely Marigold (Tagestes erecta) Shoe
attracted the attention of aquaculturist since the time
flower (Hibiscus rosasiensis) and Ixora (Ixora coccinea) in diets
immemorial. Pigments are responsible for the wide spectrum
for the sword tail (Xiphophorus helleri, Heckel, 1848). Nutrition
of colours in fishes which is an essential pre-requisite for the
is the most important factor in keeping ornamental fishes
quality as they fetch higher price in the commercial market.
healthy. In order to develop efficient and economical feed
Varieties of colouring agents are used in aqua industry to impart
formulas for aquaculture, basic information of nutrient
colour from the muscle and skin of fishes. Thus, pigmentation
requirement and chemical composition of feed ingredients in
is one of the important criterions for fishes, since their colour
relation to their acceptability and the ability of fish to digest
affect commercial acceptability.
and utilise nutrients from various sources is required. Thus, it
Carotenoids are the primary source of the pigmentation on the is necessary to conduct research and development work to
skin of fishes2. In natural environment, the fishes meet their develop effective diets which are of lower cost for the benefit of
carotenoid requirements by ingesting aquatic plants or through farmers and the industry.
their food chain. But fishes can not synthesis the carotenoid
NAAS Rating (2016)-4.20
 COLOUR ENHANCEMENT OF SWORD TAIL (284)

Table-1. ANOVA Table for fortnightly Data of Water quality parameters comparing different Days
and treatments

*All the water quality parameters shows significant difference at 0.05 level of significance (P<0.05)
Table-2. Multiple comparisons for Testing the Homogeneity between different Days
(Post HocTest using Tukey's HSD)

The mean with same superscript belongs to the homogeneous subgroup

Table-3. Multiple comparisons for Testing the Homogeneity between different Treatments
(Post Hoc Test using Tukey's HSD)

The mean with same superscript belongs to the homogeneous subgroup

(285) GOLANDAJ, SWAIN AND MOGALEKAR

Table-4. ANOVA Table for fortnightly Data of Photoshop analysis comparing different Days,
treatments Colour

* All the water quality parameters shows significant difference at 0.05 level of significance (P<0.05)
Table-5. Multiple comparisons for Testing the Homogeneity between different Days (Post HocTest using
Tukey's HSD)

The mean with same superscript belongs to the homogeneous subgroup

Table-6. Multiple comparisons for Testing the Homogeneity between different Treatments (Post HocTest using
Tukey's HSD)

The mean with same superscript belongs to the homogeneous subgroup

Table-7. Multiple comparisons for Testing the Homogeneity between different colours (Post HocTest using
Tukey's HSD)

The mean with same superscript belongs to the homogeneous subgroup

MATERIAL AND METHODS
The duration of the study was 90 days; the fishes were procured
locally and were transported under oxygen packing in polythene
bags of 20 liter capacity. The experiment was conducted
indoors in fiber glass tanks of 45 liter capacity. The fishes
were acclimatized to the culture conditions for a period of two
weeks prior to commencement of the experiment. During this
period, the fishes were maintained in an oval, flat bottom fiber
glass tank of 500 liter capacity and were fed with commercially
available pelleted feed. The tanks were filled to half of its volume
with freshwater and gentle aeration was provided. Water was
not exchanged during this period. After 14 days, the fishes
were weighed individually and randomly distributed to the
treatment tanks at the rate of 10 numbers per tank.
Three ornamental flowers namely marigold, Hibiscus and Ixora
were selected for assessment of their dietary colour
enhancement potential in the swordtail. The flowers were dried
in the shade and powdered. The dried flower petal meal was
 COLOUR ENHANCEMENT OF SWORD TAIL (286)

Figure-1 . Percentage survival obtained in different treatments

Figure-2.Specific growth rate (%) obtained in different treatments

Figure-3. Feed conversion ratio obtained in different treatments

(287) GOLANDAJ, SWAIN AND MOGALEKAR
incorporated in the diets. A basal feed without any pigment
source added served as the control. The diets were designated
as T0, T1, T2 and T3. T1 diet contained 5% of marigold petal
meal, T2 contained 5% hibiscus powder petal meal while diet
T3 contained 5% ixora petal meal. For preparation of the test
diets, the petal meal was added to the powdered control diet
@ 5%, mixed thoroughly, made into smooth dough with
sufficient quantity of water, re-pelletized and dried at a
temperature of 600C to a moisture content of less than 10%
and stored in air tight containers.
Feeding was done with ad-libitum twice in a day. Each feed
was tried in triplicate. Sampling was done at fortnightly intervals
to assess colour enhancement, fish growth and variation in
water quality parameters over the experimental period. Standard
procedures were followed for assessment of water quality
parameters on termination of the experiment-4, all surviving
fishes were collected and the length and weight recorded,
colour enhancement was assessed by calorimetrically as well
as Adobe Photoshop® version 7 Analysis.
SGR was calculated using the following formula:
ln (W2) ln (W1)
SGR (%)  100
Time interval in days
The survival rate of fishes is expressed in terms of percentage.
This was calculated as follows:
Final Number
Survival (%)  100
Initial Number
FCR was calculated by dividing the total feed intake by net
gain weight:
Total feed Intake(g)
FCR 
Net Biomass gain(g)
Photo Imaging Study: Photographs were taken according
to standard methods5. The photos were captured using a 22
megapixel digital camera. The camera was mounted on a tripod
between the two light sources fixed at an equal distance. The
camera was set up at 25 cm above the specimen and could
capture the whole fish image. All images were recorded under
uniform lighting and angle of exposure. The color parameters
of each image were analyzed with Adobe Photoshop 7 software.
Pictures were assessed at RGB mode and RGB colour profile.
Spectrophotometer Analysis: The method used for pigment
extraction from the red swordtail tissue. One gram of entire
red swordtail body tissue (without head and alimentary canal)
was taken in a 10 ml screw capped clear glass vials and 2.5 g
of anhydrous sodium sulphate was added. The sample was
gently mashed with a glass rod against the side of the vial and
then 5 ml of Chloroform was added and left overnight at 0°C.
When the chloroform formed a clear 1-2 cm layer above the
caked residue, the optical density was read at 380, 450, 470
and 500 nm, in a spectrophotometer taking 0.3 ml aliquots of
chloroform diluted to 3 ml with absolute ethanol. A blank
prepared in a similar manner was used for comparison. The
wavelength, at which maximum absorption was obtained, was
used for the calculation.
The experiment was designed in a complete randomized design
with three replications for each treatment. The average
fortnightly gain in colour and water quality parameter for each
treatment was subjected to two way ANOVA using a factorial
CRD 7 X 4 X 3.
RESULTS AND DISCUSSION
The survival rate of the fishes was in the range of 86.66-100%
in different treatments. T0 showed the highest revival rate
(100%), where T2 showed the lowest (86.66%). In the present
study the mean survival rate recorded were 100, 96.66, 86.66,
and 96.66, in T0, T1, T2, and T3 respectively. The specific
growth rate was in the range of 1.16-1.19% in different
treatments. Fish fed the T0 diet recorded the highest average
specific growth rate on termination of the study. The feed
conversion ratio recorded in the different treatments ranged
from 1.81 - 1.56 in different treatments. T2 showed the best
feed conversion ratio, where T1 recorded the poorest FCR of
1.81.
Colouration is one of the major factors deciding the market
value of the ornamental fish can be improved through dietary
enrichment using natural or synthetic pigments. Synthetic
carotenoids though effective, are often expensive. To overcome
these problems some earlier workers planned an experiment
with the cheap pigmenting source namely the marigold petal
meal6. However, the studies in this area are limited and hence
the present study was carried out to determine the potential of
Marigold, Hibiscus and Ixora as pigment enhancing agents.
In the present study, the three flower petal meals were included
at 5% level of incorporation each, in the test diets. It was
observed that fish fed all the three test diets recorded
significantly higher carotenoid content as compared to the
control fish. The highest value was recorded on the T1 diet,
followed by T3 and T2 treatments. The lowest carotenoid content
 COLOUR ENHANCEMENT OF SWORD TAIL
was in the control fish. The total carotenoid content in fishes
fed the 5% marigold petal meal diets was significantly different
from the control based on both the Photoshop analysis and
Spectrophotometer analysis. The carotenoid content in fish
fed the marigold petal meal diet was found to be 8.51 μ g/g
wet weight and 4.57 μ g/g wet weight in the control group.
Similarly, the carotenoid content in fish fed the different
experimental diets also increased significantly. The mean
weight of the sword tail (Xiphophorus helleri) on initiation of
the study was 0.72 gm in the control (T0). The initial mean
weight of fish fed the marigold 5%, Hibiscus 5% and Ixora 5%
diets were 0.78, 0.71 and 0.75 g respectively. At the end of the
experiment i.e. on the 90th day, the mean weight of the control
fishes was found to be 2.12 g, while it was observed to be
2.24, 2.04 and 2.15 g in fish fed Marigold 5%, Hibiscus 5%
and Ixora 5% diets, respectively.
The potentail of plant pigment sources for colour enhancement
in ornamentals has been studied by several workers. Some
workers studied the effect of 3 botanical additives (Coriander,
Mint and Amaranth leaves) on the growth and body colouration
of Gold fish2. The experiment was conducted in adult gold fish
for a period of 60 days. The pigment sources were added to
the basal diet at 1, 3 and 5 percent levels respectively. It was
observed that, amaranth and mint at 1 percent level enhanced
the growth and colouration of adult gold fish. In the present
investigation, the FCR varied from 1.78 to 1.57. The inclusion
of pigment sources improved the FCR as compared to the
control diet. It can be assumed that the feeds which had
pigment sources of plant origin were utilized more efficiently
by the fishes.
During the present study, the water quality parameters were
well within the ideal range suitable for the culture of warm
water species7. Inclusion of colour enhancers of plant origin
had no adverse effect on survival. On the other hand, some
researchers observed that Rosa rubiginosa had a negative
influence on the survival of Xiphophorus helleri when used for
enhancing the pigmentation in fishes through diet8. The water
quality parameters measured in different treatments in
laboratory conditions during the experimental period registered
a more or less similar trend and all of them are in acceptable
(288)
range for fish culture. The average temperature in all the
replicates varied between 25.8ºC to 27.4ºC. The average pH
values were slightly towards acidic in all the (6.56-7.66). The
dissolved oxygen content of the experimental units varied
between 4.7 to 6.1 mg/l all through the experimental period.
The total alkalinity of the water in the experimental units ranged
between 81 to 115 mg CaCo3/l throughout the period of
investigation. The ammonia value of all treatments ranged
between 0.00 to 1.16 mg/l. Nitrite values showed between
0.06 to 1.83 mg/I. Nitrate values ranged between 0.00 to 3.3
mg/l throughout the period of investigation. Some workers
suggests the ideal water quality parameter like water
temperature 26.2±0.8°C; dissolved oxygen 6.30±1.2 mg/l; pH
7.8±0.32 and total alkalinity 251±9 mg/l are considered to be
ideal for warm water fishes7. The physico-chemical parameters
observed during the study period indicates at the desired level
without much variation and is at parameter with the earlier
findings.
REFERENCES
1. Swain, S.K. and Das, B.K. (2004). In: Ornamental fish breeding
and culture: A successful economic enterprise in technologies
on livestock and fisheries for poverty alleviation in SAARC
countries "Ed. M. Abdullah, SAARC Agricultural Information
Centre "(SAICE), Dhaka, Bangladesh.
2. Ahilan, B., Jegan, K., Felix, N. and Ravaneswaran, K. (2008).
Tamil Nadu J.Veterinary and Ani. Scie.,4(4):129.
3. Meyers, S.P. (1994). Pure Applied Chem.1994; 66: 1069.
4. APHA. (1985). In: Standard Methods for the Examination of
Water and Wastewater. 18 th., Edn. American Public Health
Association, Washington DC. Aquaculture 17: 63.
5. Tlusty, M. and Hyland, C. (2005). Mar. Biol., 147: 113.
6. Ezhil, J., Jeyanthi, C. and Narayanan, M. (2008). Turkish Jour.
of Fisheries and Aqu. Scie., 8:99.
7. Boyd, C.E. (1998). In: Water Quality for Pond Aquaculture.
Research and development, International Centre for aquaculture
and aquatic Environments. Alabama Agricultural Experiment
Station. Auburn University.
8. Arulvasu, C., Ramya, M.S., Chandhirasekar, D. and Sivaganam,
S. (2013). Europ. Jour. of Biol. Scie., 5 (2): 44.