Evaluation of growth performance and breeding habits of fighting fish (Betta splendens) under 3 diets and shelters

Abstract

There are various factors affecting the fighting fish larval survival including nutrition, substrate, water quality, pathogen and stress. Experiments were conducted to study the effect of substrate and live feed on larval survival of fighting fish fry. Nine small glass aquaria (60 cm × 30 cm × 30 cm) were used as breeding tanks. Equal pieces (80 cm 2) of rigifoam (T 1), polythene (T 2) and banana leaf (T 3) were placed on the tank surface to facilitate the male to form the bubble nest under it. Thirty days of feeding trial was conducted with three different live feeds (Artemia, Moina and Bread worm) as treatments with triplicates. The bubble nest area was significantly wider (p<0.05) under rigifoam (71.85±2.02 cm 2) and polythene (32.45±1.36 cm 2) than banana leaf (24.10±1.62 cm 2). Survival of fry was significantly higher under banana leaves (96%) and rigifoam (95%) than polythene (94%). Feeding trial, final lengths of fry were 1.65, 1.23 and 1.07cm for Artemia, Bread worm and Moina respectively whereas final weights were 0.05, 0.03 and 0.02g.Better survival rates were observed among treatments with no significant difference (p>0.05).Water quality parameters recorded in different treatments during the experimental period were pH (7-7.5), temperature (26-28 0 C), and dissolved oxygen (5mg/L).

Full text

Journal of Survey in Fisheries Sciences 3(2) 50-65 2017
Evaluation of growth performance and breeding habits of
fighting fish (Betta splendens) under 3 diets and shelters
Srikrishnan R.1; Hirimuthugoda N.2*; Rajapakshe W.3
Received: March 2016 Accepted: July 2016
Abstract
There are various factors affecting the fighting fish larval survival including nutrition,
substrate, water quality, pathogen and stress. Experiments were conducted to study the
effect of substrate and live feed on larval survival of fighting fish fry. Nine small glass
aquaria (60 cm × 30 cm × 30 cm) were used as breeding tanks. Equal pieces (80 cm
2
)
of rigifoam (T
1
), polythene (T
2
) and banana leaf (T
3
) were placed on the tank surface to
facilitate the male to form the bubble nest under it. Thirty days of feeding trial was
conducted with three different live feeds (Artemia, Moina and Bread worm) as
treatments with triplicates. The bubble nest area was significantly wider (p<0.05) under
rigifoam (71.85±2.02 cm
2
) and polythene (32.45±1.36 cm
2
) than banana leaf
(24.10±1.62 cm
2
). Survival of fry was significantly higher under banana leaves (96%)
and rigifoam (95%) than polythene (94%). Feeding trial, final lengths of fry were 1.65,
1.23 and 1.07cm for Artemia, Bread worm and Moina respectively whereas final
weights were 0.05, 0.03 and 0.02g.Better survival rates were observed among
treatments with no significant difference (p>0.05).Water quality parameters recorded in
different treatments during the experimental period were pH (7-7.5), temperature (26-
28
0
C), and dissolved oxygen (5mg/L).
Keywords: Betta splendens, Substrate, Live feed, Survival Introduction
1- Department of Animal Science, Faculty of Agriculture, University of Jaffna, Sri Lanka
2- National Aquatic Resources Research and Development Agency, Rekawa, Sri Lanka
3- Department of Animal Science, Faculty of Agriculture, University of Ruhuna, Sri Lanka
* Corresponding author's Email: nyhirimuthugoda@yahoo.com

51 Srikrishnan et al., Evaluation of growth performance and breeding habits of fighting fish …
Introduction
Every year, ornamental fish demand
increase worldwide. This clearly
demonstrated by high rate of exports
come from ornamental fish producing
countries, which leads at present export
market reached 11.6% in 2006
(Fishstat, 2011). Sri Lanka ornamental
fish industry stared as commercial
industry in 1952.Now it is a thriving
industry with in export market, earning
profit and employment opportunities for
people (Kuruppan, 1998). Sri Lanka
export fresh water, brackish water,
marine water ornamental fish species
and marine invertebrates. Fresh water
ornamental fish species comprise more
colorful, more attractive and striking
fish species such as guppies, platy,
mollies, swordtail, tetras, angels,
gourami and fighter fish. Sri Lanka,
Fighting fish is a highly demanded
fresh water most beautiful, best-selling
and popular ornamental fish species
from the gourami family which is
popular as an aquarium fish. The
growth rate of Sri Lanka’s aquarium
fish industry is a healthy 4.7% with the
share of 2.7% of the US$ 412 million
world market.
Although successful breeding
techniques have been developed for this
species, high rate of mortality at the
larval production stage is a critical
issue. Fish larvae are usually very
small, extremely fragile, and generally
not physiologically fully developed
(Laven and Sorgeloos, 1996).
Combination of factors including
nutrition, substrate, water quality,
pathogen and stress decide the larval
survival (Herath and Atapaththu,
2012).The breeding method of gourami
family fish(B. splendens)is formation of
bubble nest under floating substrate.
Substrates are floating materials which
are dominant in spawning area for
construct the bubble nest.Bubble nests
are aerated place for fertilized eggs in
order to hatch those eggs successfully.
Quantity and quality of feeds are one of
the other important affecting factors for
growth performance and breeding of
ornamental fish (Degani and Yehuda,
1996). In term of larval nutrition, many
studies revealed that live feeds had a
better growth and survival than
formulated feed/Artificial feed. Within
Live feeds, Artemia has
extensivelybeen used in fighting fish
larval rearing in Sri Lanka as the sole
source of live feed since it ensures high
growth and survival. But, high cost and
occasional scarcity of Artemia makes it
unsuitable for commercial aquaculture
(Lim et al., 2003).
In recent years, there has been an
increase in the interest of B.splendens
in aquariums in Sri Lanka. High cost of
the feeds and Selection of appropriate
substrate for breeding were considered
as the major constraint for fighting fish
growers. There are scarcity of
researches about selection of a suitable
nutritionally balanced live diet (instead
of Artemia) and substrate combination

Journal of Survey in Fisheries Sciences 3(2) 2017 52
in relation to growth performance and
breeding of fighter fish. These above
issues leads to provide suitable
alternative feed for Artemia and
substrate combination in Sri Lankan
conditions become crucial for lowering
the larval mortality and sustaining the
production of ornamental fish
especially of fighting fish.Therefore,
this study is aimed to evaluate the effect
of 3 different live feeds and substrate
on the growth and breeding
performance and breeding of Fighter
fish in order to select suitable feed and
substrate which is disease free, provide
good complement of nutrients and gives
better growth and breeding
performance.
Review of literature
The Siamese fighting fish (B. splendens
Regan, 1910), widely distributed
throughout Southeast Asia, and is one
of the most popular species of great
commercial value for freshwater
aquarium. B. splendens stands out as
one of the most beautiful and best-
selling ornamental fish from tropical
water (Chapman et al., 1997; Lima,
2003). In recent years, there has been
an increase in the interest of
B.splendens in aquariums in Sri Lanka.
Moreover long-finned males are very
important economically, providing the
highest income among the exported
ornamental fishes (Wiwatchaisaet,
2000).
Larval rearing is one of the key steps
for any aquaculture achievement (Rinna
Hamlin et al., 2014). But high rate of
mortality at the larval production stage
is a critical issue for fighter fish rearing.
Special awareness should be taken for
larval rearing because this stage is more
pivotal than any other stages of life
cycle (Rinna Hamlin et al., 2014).
Fighter fish larvae are usually very
small, extremely fragile, and generally
not physiologically fully developed
(Laven and Sorgeloos, 1996).
Combination of factors including
nutrition, Shelter, Water quality,
Pathogens and stress decide the larval
survival.
Fish that build and guard bubble nests
are gourami (including Betta species)
(Jaroensutasinee, 2005). Bubble nest
building is the first step of the fish
reproductive cycle (Hall, 1968). Bubble
nests, also called foam nests, where
males build bubble nests by blowing
bubbles around a floating base. Females
spawn under the nest, where the male
waits to fertilize the eggs and typically
400-500 eggs are spawned by female
fish (Sterba, 1983; Sakurai et al., 1992).
Reproductive success of male fish
mainly depends on size of bubble nest
as it possible (Haung and Cheng, 2006).
A larger bubble nest support more
fertilized eggs and facilitates efficient
oxygenation for those eggs and larvae
(Jaroensutasinee, 2001).
In term of larval nutrition, many studies
revealed that live feeds had a better
growth and survival than that of larval
fed dry feed. Most aquarium fish are
unwilling to accept dried or flake foods
therefore live food is occasionally
necessary (Biokani et al., 2014).

53 Srikrishnan et al., Evaluation of growth performance and breeding habits of fighting fish …
Artemia also represent the only
practical food source for the early
stages of many fish and crustacean
larvae (Bardach, 1972). Siamese
fighting fish is omnivorous as
evidenced by aquarium specimens
which will readily take most food
provided. The quantity and quality of
feeds are important factors affecting the
Growth performance and reproduction
in ornamental fish (Degani and Yehuda,
1996). So, proper live feed selection is
important in early part of the growth
stage of fish.
Materials and methods
Profile of the study area
The experiment was conducted in
National aquatic resources research and
development agency (NARA) in
Rekawa Sri Lanka. All preparation
needed for breeding of fighter fish was
done in the laboratory. Aquarium set
up, water supply facilities, working
space were assured before the breeding
program. The study period was January
2015 to April 2015.
Material and Instruments
Material: Fighter fish fingerling stock
(Male and Female), breeding tanks,
substrates for breeding, fighter fish
feeds (live feed, formulated/commercial
feed), aquatic plants, Oxygen
Instruments: Electric balance, pH
meter, Dissolved oxygen meter,
Thermometer, Ruler, Oven, and all
other necessary standard laboratory
equipment were used as
instrumentsemitter pump, oxygen
stones
Methods
An experiment was conducted to study
the effect of substrate on larval survival
of fighter fish fry. Nine small glass
aquaria (60 cm × 30 cm × 30 cm) were
used as breeding tanks. One male fish
was paired with female fish in each
tank. Equal pieces (80cm
2)
of regiform,
polyethene and banana leaf were placed
on the tank water surface to facilitate
the male to form the bubble nest under
it. Different substrates such as
regifoam, polyethene and banana leaf
were considered as treatments T
1
, T
2
and T
3
. The bubble nest area was
measured by ruler and number of fry
Survive was counted. Water quality
parameters recorded in different
treatments during the experimental
period.
The second experiment was thirty days
feeding trial.That was conducted to
evaluate the growth performance of
larval fighting fish fed on three
different live feeds (T1: Artemia, T2:
Moinaand T3: Bread worm) as
treatments with triplicates. Five day old
fry of fighting fish were stocked in
glass aquaria (60 cm ×30 cm ×30cm) at
a stocking density of 25 larvae /tank
and fed on 3% of the body weight per
day and sampled in every two weeks
interval. Daily weight gain (DWG - mg
day
-1
), Specific growth rate (SGR %),
Survivalrate (SR%) were computed.

Journal of Survey in Fisheries Sciences 3(2) 2017 54
Statistical analysis
All the statistical analysis was
performed using the software SPSS
(Version 16.0) and the significance of
the difference between means was
determined using Duncan’s multiple
range test. One-way analysis of
variance was applied to determine the
significant effect of different treatments
on feeding parameters in B. splendens.
The data were expressed by mean±
standard deviation and differences were
considered significant at p<0.05. In
order to see the relationship among the
parameters, scattered plot techniques
were used.
Results
Water quality parameters recorded in
different treatments during the
experimental period were pH (7-7.5),
temperature (26-28
0
C), and dissolved
oxygen (5mg/L).In aquariums with
different color substrate, B.splendens
prefers low light intensity for the
construction of bubble nests. In this
experiment, B.splendens has more
significantly preferred to white color
rigifoam than other color substrates,
Moderate preference was observed
under the green plant leaf. Lowest
preference was observed for bubble
nest construction under grey
polyethene.
Moreover bubble nest area was
significantly wider under the regifoam
(71.85±2.02 cm
2
) and polyethene
(32.45±1.36 cm
2
) treatments than that
under the treatment with banana leaf
(24.10±1.62 cm
2
) (DMRT:
p<0.05).Survival of fry was
significantly higher under banana leaf
(96%) and rigifoam (95%) than
polyethene (94%). But larval survival
of fighting fish did not significantly
vary between the treatments of
rigifoam, polyethene and banana leaf
(p>0.05).
Larger healthier males form bubble nest
continuously which facilitate to reduce
daily decrease in size of bubble nest.
But in the present study revealed, larger
male don’t construct large area bubble
nest. There was weak positive (R=0.39)
relationship between male body weight
and construction of bubble nest area.
But In the present study of B.splendens
was able to construct bubble nest in a
wide temperature range of (25-29
o
C).
There was a strong positive correlation
(R=0.88) was observed between the
temperature and bubble nest area. The
best temperature was 28
o
C for bubble
nest construction. According to the
results, Regifoam was the better
substrate than polythene and banana
leaves in terms of bubble nest area and
fry survival. Therefore it can be
concluded that rigifoam can be used as
a better substrate to facilitate good
breeding performance and reduce the
fry mortality of fighting fish.

55 Srikrishnan et al., Evaluation of growth performance and breeding habits of fighting fish …
Table 1: Bubble nest characteristics and survival rate of Betta splendens when allow for
reproduction under three different substrates such as banana leaves, polythene and
regifoam in glass tanks during experiment-01.
Each value is a mean ± SD of three replicate, Mean values with different superscript letters in the same
row indicate significant difference at (p<0.05) (one way ANOVA and subsequently post hoc multiple
comparison with DMRT).
Figure 1: Different color substrate and preference percentage.
Figure 2: Different substrate and bubble nest area
.
Aspects Parameters Treatments
Banana leaves Polythene Regifoam
Bubble nest
characteristics
Bubble nest area (cm2) 24.10±1.62
c
32.45±1.36
b
71.85±2.02
a
Bubble nest diameter
(mm)
2.17±0.57
a
1.17±0.29
b
2.83±0.58
a
Bubble nest depth (mm) 4.17±0.58
a
2.83±0.58
b
2.83±0.57
b
Survival Survival rate (%) 96.12±2.95
a
94.15±1.30
a
95.22±3.40
a

Journal of Survey in Fisheries Sciences 3(2) 2017 56
Figure 3: Correlation between body weight and bubble nest area.
Figure 4: Male betta fish body weight and bubble nest area.
Experiment 02
Water quality parameters regarding
feeding
pH for Artemia, Moina and Bread
worm were 8.06±0.7, 7.91±0.13 and
7.89±0.12 respectively. Mean (±SD)
values of 27.71±1.13
0
C,27.61±1.05
0
C
and 27.64±0.72
0
C were recorded for
temperature and 7.00±1.02, 6.75±1.22
and 7.56±0.22 mg/L were recorded for
DO during the period of experiment-2.
Final total length and weight was
observed, Artemia fed group was
significantly higher than other two
treatments (p<0.05) (1.65±0.21cm,
0.05±0.01g). followed by Bread worm
(1.23±0.17 cm, 0.03±0.00 g) and Moina
(1.07±0.14 cm, 0.02±0.01g). There was
significant difference observed in
weight gain also; Artemia possessed a
significantly higher WG followed by
other two treatments. SGR was
significantly higher for Artemia
(3.71 0.27) lower for Moina (2.52
0.31) and Bread worm (2.94 0.00) has
possessed a SGR in between Artemia
and Moina. SGR observed for Artemia
in the present study disagreed with the
finding of Herath and Atapaththu
(2012) in which Artemia fed fighter fish
has shown SGR of 7.04±1.36 mg/day
while Breadworm has shown higher
SGR (2.94 mg/day) than the SGR
observed for Moina (4.00±0.96) by
Herath and Atapaththu (2012).

57 Srikrishnan et al., Evaluation of growth performance and breeding habits of fighting fish …
Table 2: Growth performance of B.splendens based on Initial and final total lengths,
standard lengths and body weights, survival rate WG, SGR, FCR and FCE of
B.splendens under three different live feeds such as Artemia, Moina and Bread
worm in cement tanks during experiment-02.
Parameters Treatments
Artemia Moina Bread worm
Initial total length (cm)
0.03±0.00 0.03±0.00 0.03±0.00
Final total length (cm)
1.65±0.21
a
1.07±0.14
c
1.23±0.17
b
Initial standard length (cm)
0.25±0.00 0.25±0.00 0.25±0.00
Final standard length (cm)
1.42±0.01
a
0.87±0.17
c
1.05±0.15
b
Initial weight (g)
0.0045±0.00 0.0045±0.00 0.0045±0.00
Final weight (g)
0.05±0.01
a
0.02±0.01
c
0.03±0.00
b
Survival rate (%)
82.67±6.11
a
80.00±0.17
a
84.00±0.00
a
WG (g)
0.045±0.01
a
0.018±4.00
c
0.025±0.00
b
SGR (%)
3.71±0.27
a
2.52±0.31
c
2.94±0.00
b
FCR
1.69±0.32
c
4.19±0.92
a
2.93±0.00
b
FCE
0.61±0.11
a
0.25±0.07
c
0.34±0.00
b
Each value is a mean ± SD of three replicate, Mean values with different superscript letters in the
same row indicate significant difference at (p<0.05) (one way ANOVA and subsequently post
hoc multiple comparison with DMRT).
Feed conversion ratio (FCR) and Feed
conversion efficiency (FCE) was
significantly higher for Moina
(4.19±0.92) whereas FCE was
significantly higher for Artemia
(0.61±0.11). The FCR values observed
in the present study agree with the
findings of Mandel et al. (2010). In
case of survival, there were no
significant difference among 3
treatments. This evidence shows that
any live feed among three of this live
feeds can be used to obtain a good
survival rate almost 80-84%.
Discussion
Water quality parameters such as pH,
temperature, dissolved oxygen during
the two experimental periods were
more or less supported by the findings
(Temperature: 26-28
0
C, pH: 7-7.5, DO:
5mg/L) of Biokani et al. (2014).
Influence of temperature on
reproductive behaviors
Temperature is one of the most potent
environmental parameter that
influences reproductive biology of fish
(Herzig and Winkler, 1986). Which
favor for maintaining its testes in a
reproductively active. Initiation of
bubble-nest building by the fighter fish
indicates that the reproduction of the
fish is commencing (Tsai, 1992; Huang
et al., 1998). In our study seven male
fish out of nine formed their bubble
nest within 24 hours (77.78%).
Temperature should be 25-28
o
C for the
reproduction of B. splendens in
aquarium habitats (Reyes-Bustamante
and Ortega-Salas, 1998).The present

Journal of Survey in Fisheries Sciences 3(2) 2017 58
study revealed that although
B.splendens is able to construct bubble
nest in a wide range of temperature (25-
29
o
C).The best temperature for bubble
nest construction is 28
o
C.
Influence of floating materials on
reproductive behaviors
Floating plants are usually dominant in
spawning areas of the fish (Jan, 1994).
Plants or other substrates are necessary
to hold on water surface (Degani,
1989). In aquarium habitats; B.
splendens prefers lower intensity colors
than dark color habitat for construction
of bubble nest. Dark color was least
preferred because they transmitted the
least light. Bettas don’t need that much
of light for bubble nest construction
(Harlioglu and Serpil Yonar, 2007). In
the presence study betta male more
preferred white color regifoam for
construction of bubble nest.
Subsequently green color leaf and gray
polyethene were used as a substrate for
construction of bubble nest.
Eggs floating at the water surface
appeared more likely to develop than
those submerged 10 cm below at the
bottom of a glass jar. The behavior of
male breeders could serve this physical
function since males frequently express
bubbles to the water surface, mouth the
eggs, and place the eggs into the bubble
mass. This sequence of nest-
maintenance actions could increase the
buoyancy and, indirectly, the survival
of fertilized eggs. If the male fish want
to achieve more reproductive success, it
will maintain or enhance the size of the
nest as much it can (Bronstein, 1982).
In our study, the male fish was in good
condition with white color regifoam
they construct large bubble nest than
other two substrates. But Survival not
significantly vary between the
treatments (p>0.05).
Effect of live feed on growth and
performance of fighter fish fry and fry
survival
The quantity and quality of feeds are
important factors affecting the Growth
performance and reproduction in
ornamental fish (Degani and Yehuda,
1996). In term of larval nutrition, many
studies revealed that live feeds had a
better growth and survival than that of
fry fed dry feed. In the fighter fish
larval mortality reduction and enhance
the growth rate at early stage and
induce coloration these are induce by
proper live feed selection. So, proper
live feed selection is important in early
part of the growth stage of fish.
Survival rate was significantly higher in
Artemia subsequently to bread worm
and Moina. Size of the prey and
predator, nutritional quality, density,
physical attractiveness and mode of
presentation of food basically decide
the feed intake of fish. Generally fish
larvae are able to consume prey having
same size of their mouth but small prey
is more preferable. Live feed contain all
the balance nutrition than artificial feed.
Over the experimental period providing
a wide spectrum of live feed and
measured growth performance. Mouth
gape of fish larvae decides the

59 Srikrishnan et al., Evaluation of growth performance and breeding habits of fighting fish …
preference of different size live feeds.
Some finding revealed mixture of these
three feeds also shows the better growth
performance (Herath and Atapaththu,
2012). But correct ration mixture
should needed. Bread worms are
relatively smaller in size compare to
Artemia. Hence, bread worms could be
only suitable at the onset of larval
feeding. Moina are comparatively
larger than other live feeds and
probably reduce the feed intake leading
to the low growth and survival. The
protein content of Moina usually
averages 50% of the dry weight. Adults
normally have a higher fat content than
juveniles. The total amount of fat per
dry weight is 20-27% for adult females
and 4-6% for juveniles (Das et al.,
2012). Artemia cyst protein content
62% dry weight basis and 8% of crude
fat (Hoseini, 2002).Bread worm contain
55% of protein on dry basis Fish might
perform better if different size live
feeds provide at different stages of
larval oncogenic developments.
Therefore larval rearing methods have
to be improved further for commercial
farming of the species. This experiment
was done due to replacement of 100%
Artemia by any other nutritionally equal
live feeds. Morphometric parameters
(Weight, Total length and standard
length of fish) were significantly vary
between treatments (p<0.05) and were
significantly higher in Artemia
compared to bread worm and Moina.
Weight gain (WG)
Final total length and Body weight was
observed, Artemia fed group was
significantly (p<0.05) higher
(1.65±0.21cm, 0.05±0.01g) compare to
other two feeds. Followed by bread
worm (1.23±0.17 cm, 0.03±0.00 g) and
Moina (1.07±0.14 cm, 0.02±0.01 g).
The result disagrees with WGs
observed for Artemia, bread worms,
copepods and mixture (Herath and
Atapaththu, 2012).
Specific growth rate (SGR)
SGR was significantly higher for
Artemia (3.71 0.27) lower for Moina
(2.52 0.31)and bread worm (2.94
0.00) has possessed a SGR in between
Artemia and Moina.
SGR observed for Artema in the present
study disagreed with the finding of
Herath and Atapaththu (2012) in which
Artemia fed fighting fish shown SGR of
7.04±1.36 mg/day while bread worm
shown SGR (2.94 mg/day).
Survival
There were no significant difference in
survival between 3 treatments has
provided evidence that any live feed
among three of this live feeds can be
used to obtained a good survival rate.
Feed conversion ratio (FCR)andFeed
conversion efficiency (FCE)
FCR was significantly higher for Moina
(4.19±0.92) whereas FCE was
significantly higher for Artemia
(0.61±0.11). The FCR values observed

Journal of Survey in Fisheries Sciences 3(2) 2017 60
in the present study in agreement with
the findings of Mandel et al. (2010).
Conclusions
The present study was designed to
evaluate the Growth Performance and
Breeding of Fighter Fish under three
feeds and substrates. The conclusion
provided suitable feed and substrate
combination that gives better growth
and breeding performance. From our
Study, regifoam treatment is the best
substrate treatment than other two
treatments. The bubble nest area was
significantly higher under the regifoam
(71.85±2.02cm
2
) and polyethene
(32.45±1.36 cm
2
)substrates than banana
leaf substrate (24.10±1.62 cm
2
). Larger
bubble nest areas can handle a greater
number of fertilized eggs, provide more
efficient oxygenation and support faster
embryo development. Therefore, larger
bubble nest males are able to achieve
greater reproductive success.
So regifoam selected as best substrate
for male preference for bubble nest
building and fertilize more eggs. From
the second experiment instead of the
Artemia, Moina and Bread worm can
be used as a best source of live feed
during the larval stage of the fighting
fish.
Acknowledgement
The production of this research is a
combined effort of these people,
institution and universities.I extend a
hearty thanks to Dr. N.Y.
Hirimuthugoda, Dr. A.D.W.R.
Rajapakshe and of Dr. Miss. J.
Sinniah, Head/Animal Science, all who
contributed to the completion of this
research project.
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