Reproductive performance of seahorse, Hippocampus barbouri (Jordan and Richardson 1908) in
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Journal of Survey in Fisheries Sciences 2(2)17-33 2016
[ DOI: 10.18331/SFS2016.2.2.2
Reproductive performance of seahorse,
Hippocampus barbouri (Jordan and Richardson 1908) in
control condition
Nur F.A.H.1; Christianus A.1,2*; Muta Harah Z.2; Ching F.F. 3;
Shapawi R.3; Saad C.R.2; Senoo S.3
Received: July 2015 Accepted: December 2015
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Abstract
Hippocampus barbouri is one of the seahorse species found in shallow water of
Malaysia. It is used as known as a global trade species in ornamental fish industry. To
date, there is no documented report on seahorse aquaculture especially for H. barbouri
in Malaysia even in Southeast Asia region. Seahorse aquaculture should be considered
as an alternative source of seahorses to reduce the pressure on wild population.
Therefore, this study was conducted to establish suitable techniques for broodstock
maintenance and reproduction by focusing on culture system and feeding.
Hippocampus barbouri were maintained and bred successfully in a controlled culture
system. Minimum water depth required for the spawning of H. barbouri is 38 cm. Best
reproductive performances was observed in broodstock fed with post-larvae shrimp.
However, frozen mysid can also be used in the culture of H. barbouri. The minimal
requirement of n-3 and n-6 fatty acids for the reproduction of H. barbouri was 5.13 ±
0.04 % and 14.83 ± 0.10 % respectively.
Keywords: Hippocampus barbouri, Culture system, Feeding
1-Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
2-Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, 43400
Serdang, Selangor, Malaysia.
3-Borneo Marine Research Institute, Universiti Malaysia Sabah, 88400 Kota Kinabalu, Sabah,
Malaysia.
*Corresponding author's email: annie@upm.edu.my
18 Nur et al., Reproductive performance of seahorse, Hippocampus barbouri in…
]
[ DOI: 10.18331/SFS2016.2.2.2
Introduction in captivity is still at infancy stage with
Seahorses belong to the Syngnathidae husbandry problems and high juvenile
family includes pipefishes, pipehorses mortalities (Vincent, 1996; Planas et
and seadragons (Lourie et al., 1999). al., 2008). In the process to develop of
Harvesting seahorses to fulfill the high suitable techniques for broodstock
demands for traditional Chinese maintenance and reproduction, special
medicine (TCM) and marine aquarium focus was given to culture system and
trade resulted in the decline of the wild feeding (Planas et al., 2008). Culture
populations ( Vincent, 1996; Lourie et system is an important factor
al., 1999; Foster and Vincent, 2004). particularly to maintain water quality in
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Seahorses also an icon for issues related order to provide favorable conditions
to incidental by-catch and habitat loss for seahorse broodstock (Koldewey,
(CITES, 2002). Therefore, seahorses 2005; Planas et al., 2008).
are among the fourteen bony fishes Based on previous study, diets for
listed in Appendix II of the Convention seahorse broodstock have great
on the International Trade of influence on gonad development and
Endangered Species of Wild Fauna and brood size (Wong and Benzie, 2003;
Flora (CITES, 2002) and Red List of Sheng et al., 2006). Adult Artemia were
International Union for Conservation of regularly given to seahorse broodstock
Nature as Threatened Species (IUCN, in captivity compared to other variety
2006). of food items, including mysid shrimp,
Ten species of seahorses were found shrimps and amphipods either as live or
in Malaysia, this includes Hippocampus frozen diet (Woods and Valentino,
barbouri (Lim et al., 2011). Its shallow 2003; Dzyuba et al., 2006; Lin et al.,
water habitat in Malaysia exposed this 2006; Lin et al., 2007; Buen-Ursua et
species to human activities (Choo and al., 2015). Providing a diet that fulfill
Liew, 2004). As a global trade species, nutritional requirement is considered as
H. barbouri is the most common a great challenge in seahorse
species kept in aquarium due to its aquaculture.
uniqueness in appearance and attractive Success reproduction is crucial to
color variation from white, pale yellow ensure the sustainability of seahorse
to pale brown (Kuiter, 2000; Koldewey aquaculture (Payne, 2003; Lin et al.,
and Martin-Smith, 2010; Olivotto et 2007). To date, no report on seahorse
al., 2011). aquaculture in Malaysia and the
Seahorse aquaculture can be Southeast Asian region (Koldewey and
considered as an alternative source in Martin-Smith, 2010). Therefore, this
order to reduce the pressure on wild study was conducted to establish
population (Payne and Rippingale, breeding technology for H. barbouri
2000; Lin et al., 2007; Faleiro et al., which will contribute to the
2008). However, production of seahorse development of seahorse aquaculture.]
Journal of Survey in Fisheries Sciences 2(2) 2016 19
[ DOI: 10.18331/SFS2016.2.2.2
Materials and methods barbouri. Each tank was equipped with
Acquisitions of seahorse Classica® Crystal Hang-on Filter (EF-
Adult seahorses were bought from 087). Sponge and sintered glass
fishermen at Semporna coast of Sabah, Biohome® Plus were used as filter
Malaysia. These seahorses were media. In IBS, rectangular fiberglass
conditioned for 1 month before being tanks (0.4×0.4×0.5m) were used with
used for breeding trials. Tilapia fry and setup similar to BMRI. Glass tanks
shrimp postlarvae were used as feed (0.4L×0.3W×0.4H m) were used as
during this conditioning period. Only breeding tank in IBS. Each of these
healthy seahorses were used for all tanks was equipped with hang-on filter
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experiments. (with sponge and Biohome as filter
media), similar to the setup at DoA.
Culture system Figure 1, 2 and 3 shows the front view
Breeding experiments were conducted of both conditioning and breeding tank
at three different locations, Borneo set up in BMRI, DoA and IBS
Marine Research Institute, Universiti respectively. In each conditioning and
Malaysia Sabah, Kota Kinabalu, Sabah, breeding tank, plastic chain tied to a
Malaysia (BMRI); Department of sinker served as holdfast for seahorses.
Aquaculture, Universiti Putra Malaysia, After conditioning for 1 month, each
Serdang, Selangor, Malaysia (DoA) and pair of broodstock with average
Institute of Bioscience, Universiti Putra standard length (SL) 12.040±0.459cm
Malaysia, Serdang, Selangor, Malaysia was transferred into breeding tank.
(IBS). Different culture systems were Standard length was determined by
used in each location for the measuring the length from the tip of the
conditioning and breeding of wild H. tail to the mid-point of the cleithral ring
barbouri. plus the length from the tip of the snout
In BMRI, broodstock were to the mid-point of the cleithral ring
conditioned in one tonne black circular (Lourie et al., 1999). Seahorses were
polyethylene tank (H=0.8m). This blotted using filter paper before being
conditioning tank was connected with weighed to get the wet weight, WW
another tank of similar size as a (Job et al., 2002). Data on reproductive
filtration unit filled with hard coral as performance was collected only after
filter media. Airlift technique was used two weeks of introduction into breeding
to circulate water from one tank to the tank. This experiment was carried out
other. For breeding tank, similar system for 3 months.
as conditioning tank was set using Prior to usage, seawater undergoes
square fiberglass tank (0.4×0.4×0.4m). serial filtration (cartridge pore size of 5,
In DoA, plastic tanks 1 and 0.1µm) then passed through
(0.55×0.25×0.33m) were used for both Atman® UV Sterilizer (Model UV-
conditioning and breeding of H. 11W) with water flow rate of 10 l/min.20 Nur et al., Reproductive performance of seahorse, Hippocampus barbouri in…
]
[ DOI: 10.18331/SFS2016.2.2.2
Broodstock was fed twice daily, ad twice a week. Food consumption in the
libitum with frozen Hikari® Bio-Pure percentage of body weight (% BW) was
Mysis Shrimp at 0930 and post-larvae estimated by measuring the amount of
of white shrimp and red tilapia fry at food intake per total weight of seahorse
1530. Faeces and excess feed were individual per day
siphoned daily before and after feeding. (individual/seahorse/day). Experiment
Water depth in the tank was maintained was carried out for three months.
throughout the experimental period. Quantity of diets fed to seahorse was
YSI Professional Plus Multi-Parameter estimated based on the data of food
and HACH® DR/2400 Portable consumption. Throughout this study,
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Spectrophotometer were used to water quality was monitored and data
measure dissolved oxygen, DO (ppm), on reproductive performance were
temperature (°C), salinity (ppt), pH, recorded.
ammonia (ppm), nitrate (ppm), and
nitrite (ppm) twice a week throughout Diet preparation
this experiment. Newly hatched Artemia nauplii from
Bio Marine Artemia cysts were grown
Feeding experiment in circular fiberglass tank with vigorous
Experiment conducted at IBS used aeration at stocking density 200/l.
similar breeding tank set up as in the Artemia nauplii were fed with rice flour
previous experiment. After one month and Spirulina powder (Josens) until
conditioning, each pair of broodstock reach adult stage (total length, TL: 5-
with average SL and WW 10mm). Adult Artemia was enriched
(12.100±0.424cm and 5.551±0.322g) with cod liver oil for twelve hours prior
were introduced into each breeding to feeding.
tank. Broodstock were fed with adult Both PLS and TF bought from fish
Artemia (AA), frozen Hikari® Bio-Pure farms once a month were maintained in
Mysis Shrimp (FM), post-larvae of one tonne rectangular fiberglass tank.
white shrimp (PLS) and fry of red The average TL of PLS (the tip of
tilapia (TF) twice daily at 0930 and rostrum to the tip of telson) and TF (the
1530. tip of closed mouth to the extended tip
Diets were rinsed few times with of the caudal fin) that were fed to
freshwater and blotted using filter paper seahorse broodstock were 13±2mm and
to measure the WW of the diets prior 6±2mm respectively. Prior to use, TF
fed to the seahorse broodstock. After an fry fed with newly hatched Artemia
hour of feeding, excess diets were nauplii ad libitum for five minutes.
siphoned, blotted using filter paper and Freshwater was used to defrost and
weighted to determine daily food intake clean FM. All diet were rinsed with
of seahorses broodstock. Sampling for filtered freshwater and seawater for 3
weight of broodstock was conducted times before being fed to the]
Journal of Survey in Fisheries Sciences 2(2) 2016 21
[ DOI: 10.18331/SFS2016.2.2.2
broodstock. Samples of three diets were yolk sacs was estimated based on length
(L) and width (W) measures, using the
stored at -20°C for proximate analysis 1
volume of an ovoid, V= ×πLWW
and fatty acid profiling.
6
(Faleiro et al., 2008).
Data collection
Data collected were used to calculate
Proximate Analysis
the weight gain (WG) and specific
Proximate analysis (protein, lipid, ash
growth rate (SGR) during the
and moisture) of diet samples were
experiment. WG(%)=(Wf−Wi)/Wi×100
conducted according to standard
and SGR(%)=((lnWf−lnWi)/t)×100
methods AOAC.
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where, Wf is the final weight (g), Wi is
the initial weight (g), SL is the standard
Fatty acid profiling
length (cm), and t is the duration (d).
Fatty acid profiling was carried out
To measure the reproductive
respectively after total lipid
performance of seahorse, data on the
determination. Lipid from sample was
occurrence of spawning, unsuccessful
extracted using Soxhlet extraction
spawning (when all of unfertilized eggs
apparatus. Fatty acid methyl esters
found at tank bottom), pregnancy
(FAME) were injected into a capillary
period (number of days male incubated
column SUPELCO SP-2380 (100m
the fertilized eggs until giving birth),
fused silica, 0.25mm internal diameters)
brood size (number of newborn juvenile
installed in a HP-5890 Series II Plus gas
seahorse), size of newborn juvenile (SL
chromatograph. Peaks were identified
and WW), number of unfertilized eggs
by comparison with standard FAME as
(from unsuccessful spawning or some
an internal standard.
of unfertilized eggs aborted during
spawning), number of aborted eggs
Data analysis
(fertilized eggs being aborted from
Data were presented as mean ± standard
brood pouch during pregnancy) and
deviation (SD). Data collected during
number of premature juvenile were
the feeding experiment were analysed
recorded. The newborn juvenile
using one way of Analysis of Variance
seahorses were cultured for two weeks.
(ANOVA) and Tukey test to determine
Survival was recorded to determine the
the significant difference between the
effect of broodstock diet on juvenile
treatments. Correlation coefficient of
quality.
reproductive performance to selected
Unfertilized eggs found at the bottom
biochemical parameters were analyzed
of the tanks were siphoned and counted.
using correlation test at a=0.05. All
Eggs diameter was measured using
statistical analyses were carried out
ocular micrometer fits in eyepiece of
using SAS 9.4
Leica® DM500 Compound Microscope.
Volume of the unfertilized eggs and22 Nur et al., Reproductive performance of seahorse, Hippocampus barbouri in…
]
[ DOI: 10.18331/SFS2016.2.2.2
Results set up and data on reproductive
Culture system performance between different
Different culture systems used at locations. Besides the variation of tank
different location depend on the set up, feed may influence the
availability of materials (tank, filter, reproductive performance of seahorse.
filter media). In this study, adult H. Therefore, the next experiment was
barbouri were maintained and bred in conducted to determine the actual
captive conditions using different effects of provided diet on the
culture system. However, gas bubble reproductive of H. barbouri.
disease (GBD) occurs frequently in air-
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lift culture system at BMRI. Feeding experiment
Conditioning of H. barbouri in DoA Post-larvae of white shrimp (PLS) was
with high stocking density resulted in the most preferred feed since
highest level of ammonia, nitrate and broodstock of H. barbouri consumed
nitrite compared to other locations. 9.81 ± 0.77% by body weight of PLS
Table 1 shows the comparison in tank daily, which was significantly higher
set up and water quality data between (p]
Journal of Survey in Fisheries Sciences 2(2) 2016 23
[ DOI: 10.18331/SFS2016.2.2.2
was produced by broodstock fed on AA PLS produced significantly highest
(24.5±7.78 pieces) or TF (27.5±10.61 (p24 Nur et al., Reproductive performance of seahorse, Hippocampus barbouri in…
]
Table 2: Data on breeding of Hippocampus barbouri at three different locations.
[ DOI: 10.18331/SFS2016.2.2.2
BMRI DoA IBS
Water depth 0.35 m 0.30 m 0.38 m
Tank Size (m) 0.4L × 0.4W × 0.4H 0.55L × 0.25W × 0.33H 0.4L × 0.3W × 0.4H
Colour Blue Transparent Transparent
Material Fiberglass tank Plastic tank Glass tank
Filtration System Air-lift Hang-on Hang-on
Media Hard coral Sponge & Bio-home Sponge & Bio-home
Stocking density 1 ind/ 28l 1 ind/ 20l 1 ind/ 22l
Feeding frozen Hikari® Bio-Pure Mysis Shrimp, red tilapia fry & post-larvae of white shrimp
Water DO (ppm) 7.41 ± 0.16 5.25 ± 0.23 5.80 ± 0.07
parameter Temperature (°C) 27.88 ± 0.85 27.03 ± 0.24 29.17 ± 0.98
Salinity (ppt) 31.88 ± 0.38 29.83 ± 0.92 30.47 ± 0.80
pH 7.98 ± 0.15 7.76 ± 0.17 8.16 ± 0.10
Ammonia (ppm) 0.10 ± 0.04 0.13 ± 0.02 0.04 ± 0.01
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Nitrate (ppm) 1.23 ± 0.43 2.38 ± 0.42 1.58 ± 0.52
Nitrite (ppm) 0.03 ± 0.02 0.05 ± 0.02 0.02 ± 0.01
Reproductive Spawning occurrence 13 10 20
performance Unsuccessful 2 2 -
spawning
Incubation period 14-15 days 14-15 days 13-14 days
Brood size 54.77 ± 5.183 26.70 ± 9.129 59.05 ± 4.947
Unfertilized eggs 93 114 2
Juvenile size SL:0.83 ± 0.02 cm SL:0.81 ± 0.03 cm SL: 0.85 ± 0.02
WW: 0.004 ± 0.001 g WW: 0.004 ± 0.001 g WW: 0.005 ± 0.001 g
No. of premature - 23 -
Table 3: Data on food consumption, weight gain (WG) and specific growth rate (SGR) of
Hippocampus barbouri fed on different feed.
Treatment Amount of feed consumed WG (%) SGR (%)
Wet weight (g) Percentage by body weight
(%)
Adult Artemia 0.30 ± 0.01b 5.80 ± 0.14b 9.28 ± 0.28b 1.27 ± 0.04b
b b
Frozen mysids 0.32 ± 0.01 5.83 ± 0.37 12.29 ± 1.74b 1.65 ± 0.22b
a a
Post larvae shrimp 0.55 ± 0.02 9.81 ± 0.77 21.30 ± 0.67a 2.76 ± 0.08a
c c
Tilapia fry 0.23 ± 0.02 4.11 ± 0.28 10.88 ± 0.82c 1.48 ± 0.11c
Different superscript letters shows significant differences between treatment at p]
Journal of Survey in Fisheries Sciences 2(2) 2016 25
[ DOI: 10.18331/SFS2016.2.2.2
Table 5: Data on proximate composition of different feed.
Treatment Percentage by dry matter (%)
Moisture (%) Energy (kJ)
Protein Fat Carbohydrate Ash
Adult Artemia 92.73 ± 0.04a 33.28 ± 3.31c 12.31 ± 0.03b 29.03 ± 2.31a 25.38 ± 1.02a 110.91 ± 2.96d
Frozen mysids 91.42 ± 0.00b 60.14 ± 2.15b 4.72 ± 0.09c 21.56 ± 1.49b 13.58 ± 0.58b 134.00 ± 0.00c
Post larvae
87.24 ± 0.05d 66.51 ± 0.48a 3.68 ± 0.13d 11.67 ± 0.62c 18.14 ± 0.01c 185.00 ± 0.00b
Shrimp
Tilapia fry 87.84 ± 0.14c 65.60 ± 1.99ab 17.19 ± 0.31a 7.10 ± 2.07d 10.12 ± 0.23d 227.00 ± 0.00a
Different superscript letters shows significant differences between treatment at p26 Nur et al., Reproductive performance of seahorse, Hippocampus barbouri in…
]
[ DOI: 10.18331/SFS2016.2.2.2
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Figure 1: Front view of A, conditioning tank and B, breeding tank in BMRI. PVC
pipe (Ø = 5cm) and air-lift technique were used to circulate water from one tank to
another.
Figure 2: Front view of both conditioning tank and breeding tank in DoA. Hang-
on with sponge and sintered glass Biohome® Plus were used to
filtered water.]
Journal of Survey in Fisheries Sciences 2(2) 2016 27
[ DOI: 10.18331/SFS2016.2.2.2
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Figure 3: Front view of A; conditioning tank and B; breeding tank in IBS.
Discussion in air-lift system creates water
Development of suitable techniques movement as well as supplemental
especially the physical factor is very aeration to minimize the consumption
important for broodstock maintenance of resources (Loyless and Malone,
and reproduction (Planas et al., 2008). 1998). However, it causes the presence
Culture system is the physical key of air bubble in culture system and
factor to provide favourable conditions increases the occurrence GBD in
with minimal resources ( Duarte et al., seahorses (Reinemann, 1987; Planas et
2011; Blanco et al., 2014). Based on al., 2008).
this study, H. barbouri can be Super saturation of gas especially
maintained in different culture system. nitrogen and oxygen typically related
However, there are some factors with the application of air-lift technique
constraining the successful spawning of (Parker et al., 1984). This super
H. barbouri in captive conditions. saturation become the causative agent
Unsuccessful spawning in the present for GBD in seahorse, whereby gas
study frequently relates to the entrapment in the brood pouch,
occurrence of GBD in seahorse cultured subcutaneous emphysema on the tail28 Nur et al., Reproductive performance of seahorse, Hippocampus barbouri in…
]
[ DOI: 10.18331/SFS2016.2.2.2
segment or hyperinflation of swim broodstock in this study due to its ready
bladder (Koldewey, 2005; Planas et al., supply. Furthermore, Garcia et al.
2008; Koldewey and Martin-Smith, (2012) reported the use of fish larvae as
2010). During this study, air-lift system feed for adult H. barbouri.
recorded the highest dissolved oxygen, Broodstock of H. barbouri fed on
to near saturation level as suggested by PLS shows the best reproductive
Masser et al. (1999). performance with high numbers of
Tank height is another restrictive spawning occurrences and brood size.
factor to the successful breeding of H. Coincidentally, it was the most
barbouri. It is important to ensure water preferred feed by the seahorses as
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depth is sufficient to enable successful compared to the other feeds. Based on
eggs transfer to the male brood pouch optimum foraging theory, seahorse
during spawning process (Woods, prefer to consumed caridean shrimp
2003; Koldewey, 2005). Hippocampus with lowest energy expenditure
abdominalis requires tank depth of at required (Anderson Jr, 2000; Felício et
least 90 cm to facilitate the egg transfer al., 2006). In addition, structure of
(Sobolewski, 1997). However, high syngnathid eyes made them adaptable
water level will cause difficulty in tank to the mobility and carotenoid-rich prey
maintenance. Therefore, it is important (Collin and Collin, 1999). Similar to
to determine minimum water depth previous finding, adult H. barbouri
required to ensure successful eggs prefers to consumed FM compared to
transfer during seahorse spawning. the highly mobile AA and TF (Felício
Breeding tank with different water et al., 2006).
depth was tested in this study. All Quality and quantity of feed give a
seahorse broodstocks spawned major influence on brood size, which
successfully in tank with 38 cm depth affects their gonad development and
in IBS. Hence, H. barbouri required at sperm quality (Wong and Benzie, 2003;
least 38 cm water depth for successful Foster and Vincent, 2004; Lin et al.,
eggs transfer.
2007). According to Otero‐Ferrer et al.
Adequate of nutrients is one of the (2012) the utilization of live feed
main factors influencing the spawning resulted in better growth and gonad
outcome of teleost fish (Izquierdo et al., development of adult seahorses.
2001). Variety of feeds were given to However, in this study broodstock fed
seahorse broodstock in captivity which with FM showed better reproductive
include adult Artemia, mysid shrimp, performance (in terms of spawning
amphipods and shrimps, given as live occurrence, brood size and juvenile
or frozen (Woods and Valentino, 2003; survival) as compared to broodstock fed
Dzyuba et al., 2006; Lin et al., 2007; with live feed such as AA and TF. This
Palma et al., 2012). In this study, TF condition may due to the higher
was selected as feed for seahorse]
Journal of Survey in Fisheries Sciences 2(2) 2016 29
[ DOI: 10.18331/SFS2016.2.2.2
percentage of FM consumed by H. amphidromous marine fish contained
barbouri as compared to AA and TF. higher n-6 compared to n-3 (Usman,
Composition of n-3 HUFA, 2014). In East Malaysia, H. barbouri
especially DHA considered as the usually found together with H. kuda at
major dietary requirement for the estuaries or near the river mouths
successful reproduction in marine fishes (Choo and Liew, 2004). Therefore, H.
by affecting steroidogenesis, barbouri may be an amphidromous
spermiation and other reproductive marine fish that requires more n-6
parameters (Izquierdo et al., 2001). compared to n-3. Highest percentage of
DHA plays an important role in cells protein and n-6 found in PLS compared
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membrane by regulating the integrity to other feed offered in this study may
and function besides being an important likely be the reason of better
component of phosphoglycerides in reproductive performance of H.
gonad and juvenile (Izquierdo et al., barbouri.
2001). Its deficiencies can cause H. barbouri can be maintained and
decreases in fecundity, lower bred in captive condition in Malaysia.
fertilization rates, embryo deformities However, presence of bubble gas in
and poor larval quality (Izquierdo et al., culture system should be avoided since
it can be a causative agent for GBD in
2001; Otero‐Ferrer et al., 2012). In
contrast, the present study, broodstock seahorse. Minimum water depth
H. barbouri fed on PLS (which lower in required for successful spawning of H.
n-3 contents) compared to TF, shows barbouri is 38 cm. Based on this study,
better reproductive performance evident the best diet for H. barbouri broodstock
with bigger brood size with no is PLS, obvious with the best
abnormality on newborn juveniles. Low reproductive performance. However,
n-3 content in live feed may be FM can also be used in the culture of H.
sufficient to compensate the dietary barbouri. The minimal requirements of
n-3 and n-6 fatty acids for reproduction
requirement for seahorse (Otero‐Ferrer
et al., 2012). In addition, low n-3 of H. barbouri are 5.13±0.04% and
content also found in eggs of H. 14.83±0.10% respectively. Further
guttulatus (Planas et al., 2008) study on bioavailability and n-6
Composition of protein in feed also requirements for H. barbouri should be
affects the reproductive performance of conducted.
marine fish (Izquierdo et al., 2001;
Buen-Ursua et al., 2015). For example, Acknowledgement
the reduction in protein composition Author would like to thank the Ministry
reduced egg viability in seabass (Cerdá of Higher Education Malaysia for
et al., 1994). Often freshwater fish funded this study through Fundamental
contained higher n-6 compared to n-3 Research Grant Scheme (FRGS).
(Steffens, 1997). However, some30 Nur et al., Reproductive performance of seahorse, Hippocampus barbouri in…
]
[ DOI: 10.18331/SFS2016.2.2.2
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