Effects of pH, feeding regime and thyroxine on growth and survival of Parosphromenus tweediei (Kottelat and Ng, 2005) fry
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Journal of Survey in Fisheries 8(1) 89-105 2021
Effects of pH, feeding regime and thyroxine on growth and
survival of Parosphromenus tweediei (Kottelat and Ng, 2005)
fry
Zulkifle M.S.1; Christianus A.1,2*; Zulperi Z.1
Received: October 2019 Accepted: May 2020
Abstract
Parosphromenus tweediei is a rare native fish of Malaysia. It is found only in peat swamp areas.
Being traded at high market price of MYR40 per fish shows its potential as ornamental fish.
Currently, this fish is threatened to extinction due to anthropogenic activities. Particularly when
peat swamps are converted into paddy, pineapple or oil palm plantations. To date, there are very
few studies on P. tweediei. Culture of this species is non-existent. Therefore, the objectives of
this study were to look into several factors affecting the growth and survival of this species in
control condition. Aspects studied were on the effect of pH, feeding regime and thyroxine (T4) on
growth and survival of P. tweediei fry. Experiments were carried out at Hatchery Unit, Institute
of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Parameters for water
quality such as dissolved oxygen, temperature, ammonia, nitrite and nitrate were monitored once
a week throughout the experimental period. All experiments were conducted using polystyrene
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box with size of 34x21x19cm. Experiment on pH consisted of pH 4.5, 5.0, 5.5 and 6.0, which
were prepared in triplicates with stocking density of 12 fry per replicate. As for experiment on
feeding regime, three regimes used were AM: Artemia nauplii (6-20DAH) - Moina sp. (21-
63DAH), AMG1: Artemia nauplii (6-20DAH) - Moina sp. (21-43DAH) - Grindal worm (44-
63DAH) and AMG2: Artemia nauplii (6-20DAH) - Moina sp. (27-48DAH) - Grindal worm (49-
63DAH). All treatments were triplicated with 10 fry per tank. As for T4 experiment, 3 dosages
tested were 0, 0.5 and 1 ppm, conducted in duplicates with 10 fry per tank. Immersions of T4 on
28 DAH fry were carried out only once at the beginning of the experiment. Results showed that
fry of P. tweediei can be successfully cultured from pH 4.5 to 6.0. It was observed that total length
(TL) and survival increase as pH level decreases from 6.0 to 4.5. As for feeding regime, statistical
analysis showed that there were significant differences (p90 Zulkifile et al., Effects of pH, feeding regime and thyroxine on growth and survival of …
Introduction growth and survival of P. tweediei fry
Growth and development of fish are cultured at different pH.
influence by environmental factors and The availability and quality of feeds
food availability. Maintenance of water are also important factors contributing to
quality with suitable water requirements decent growth performance and
will enhance the survival and growth of successful breeding of ornamental fish
the fish. Fish continuously grow as they (Degani and Yehuda, 1996). During
live and relied on external environment nursery stage, the lack of suitable food
(Brett, 1979; Boeuf et al., 1999). The causes low survival of fry (Chaudhuri,
combination of several factors like 1979). Often fish breeders use live food
nutrition, substrate, water quality and in the culture of ornamental fish. It is use
stress affect the survival of the fish to induce ornamental fishes to breed
(Herath and Atapaththu, 2012). These (Shelar et al., 2014). Studies also
ecological factors can be divided into revealed that fishes have better growth
physical and chemical factor. Physical and survival when fed with live food
factor includes temperature, salinity, and (Srikrishan et al., 2017). Live food
photoperiod. While the chemical factor provides the required nutrients as they
encompassed ammonia and pH. are ready for consumption and efficiently
Chemical factor like pH is very digested to support growth of fish larvae
crucial in fish culture. It is an indicator of (Kumar et al., 2008). Commonly used
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acidic or alkaline condition. Water live food in ornamental fish culture is
considered acidic when pH is below 7 Artemia, Moina, copepods, Brachionus
and considered alkaline when above 7. In and Chrinomus (Herath and Atapaththu,
aquaculture, pH between 6.5 and 8.5 is 2012). In recent years, Artemia has
considered as the most suitable range and extensively used as live food since it
recommended as a guideline for proper ensures better growth and survival of fish
practice (Summerfelt, 1998). However, fry. However, Artemia is quite costly
this may not be applicable for certain (Lim et al., 2003). So, identification of
species of fish. Suitable pH conditions low cost live food is needed as to reduce
prevent metabolic stress and help fish the production cost as well as to ensure
efficiently utilized nutrients from the sustainability of the fish production.
feed and at the same time optimize Therefore, the second experiment
digestion, thereby maximizing the evaluates the effect of different feeding
growth of the fish (Bolner and regimes on growth and survival of P.
Baldisserto, 2007; Rebouças et al., tweediei.
2015). Based on their natural habitat, P. Growth hormone is important during
[ DOI: 10.18331/SFS2021.8.1.7 ]
tweediei thrive well in an acidic early life stage of fish. It plays an
condition. In order to gauge the water important role since high level of thyroid
parameters tolerance of the species, a hormone is found in eggs, and it acts as
study was conducted to compare the promoting agent for larva-juvenile
transition (Yamano, 2005). ThyroidJournal of Survey in Fisheries Sciences 8(1) 2021 91
hormone contained tri-iodothyronine addition of Java Fern, Microsorum
(T3) and prohormone thyroxine (T4). T3 pteropus attached to drift wood and 5cm
and T4 are widely used in animal diameter PVC pipe (7-12cm length). The
husbandry and play an important role in side of the tank was covered with black
the application for vertebrate, since it can plastic to reduce the penetration of light.
influence metabolism (Gorbman, 1969). Peat granules (JBL Tormec Activ) were
Several studies showed the effect of T3 soaked in the water to get the desired pH
and T4 application onto eggs and fish for breeding. All tanks were prepared one
larvae. It enhances growth and survival week before fish was placed inside. The
of tilapia, conger eel, sea bream, striped tanks were monitored in the morning and
bass, rabbit fish, coho salmon, sockeye evening in order to observe the presence
salmon and pink salmon (Power et al., of eggs. Male usually makes bubble nest
2001). As growth factor in fish is very in the inner surface of the PVC pipe.
important, this led to the determination of Upon successful spawning, eggs were
T4 effect on the growth and survival of P. kept in the breeding tank until hatched
tweediei fry. This hormone can be and at 3 days after hatch (DAH), all the
applied on fish through immersion, postlarvae attached to the inner side of
injection or incorporated into feed (Higgs the PVC pipe, were removed into the
and Eales, 1973; Eales, 1974; Hurlburt, larval rearing tanks. At this time,
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1977). postlarvae were fed with green water
taken from tilapia pond. This green water
Material and methods consisted of Monoraphidium sp.,
Location of study Closterium sp., Klebsormidium sp.,
Study was conducted at the Hatchery Chlorella sp. and Scenedesmus sp.
Unit, Institute of Biosciences, Universiti
Putra Malaysia, Serdang, Selangor,
Malaysia. Adults Parosphromenus
tweediei (Fig. 1) were caught from the
wild along roadside canal at the vicinity
of Sri Bunian, Pontian, Johor. Three
water parameters measured at the natural
habitat were pH, temperature and
dissolved oxygen.
Breeding trial
[ DOI: 10.18331/SFS2021.8.1.7 ]
Broodstocks were conditions for 3 weeks Figure 1: Adult Parosphromenus tweediei
prior to breeding. Fishes with total length
of 2.3-2.7cm, with a ratio of 1 male: 1 pH Tolerance
female was used. Breeding tank with size Culture water was prepared from
26x15x15 cm was prepared with the dechlorinated tap water into pH 4.5, 5.0,
5.5, 6.0, and prepared in triplicates with92 Zulkifile et al., Effects of pH, feeding regime and thyroxine on growth and survival of …
stocking of 12 fry per replicate. All the (Sigma Chemical, USA) was dissolved
pH values were achieved using peat in distilled water containing 0.06% NaCl
granule, JBL Tormec Activ. Seven day per liter and pH was adjusted between pH
after hatch (DAH) fry were placed in 8.0 to 8.5 with 1 N alkali and 0.1 N HCl
polystyrene box size 34x21x19cm and (Mareedu and Gudamani, 2013). Each
observed weekly until 63 DAH. Water treatment tank was duplicated with
change of 10% was carried out weekly. stocking of 10 fry per replicate. Similar
to feeding regime experiment, the pH of
Feeding regime water used in this experiment was based
Three feeding regimes used in this study on the best pH from the first experiment.
were AM, AMG1 and AMG2. The first Water changes of 10% were carried out
feeding regime AM consisted of Artemia weekly.
nauplii given from 6 DAH, then followed
by Moina sp. at 21 DAH onwards. The Total length and Survival
second feeding regime, AMG1 consisted Sampling for total length (TL) of fry was
of Artemia nauplii from 6 DAH, then carried out, once a week to obtain the
Moina sp. from 21 DAH and grindal average length. Total length was
worm, Enchytraeus buchholzi from 44 measured from the tip of the mouth to the
DAH onwards. The third feeding regime, caudal fin of the fish (Fishbase, 2004).
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AMG2 consisted of Artemia nauplii The lengths were measured using digital
given from 6 DAH, and then followed by calliper (Mitutoyo, Thailand). Mortality
Moina sp. from 28 DAH and then grindal of fry was observed and noted
worm, Enchytraeus buchholzi from 49 throughout the experimental period.
DAH onwards. Each treatment was
triplicated with stocking of 10 fry per Water quality measurements
replicate. Feeding regime experiment Water pH was measured using pH pen
was conducted after the pH experiment. (Hanna, Romania). While temperature
Therefore, the pH of culture water used (oC) and dissolved oxygen, DO (mg/L)
in this experiment was based on the best were measured using YSI Multi-
result from the pH experiment. Water Parameter. While, API freshwater master
changes of 10% were carried out weekly. test kit was used to determine the
ammonia, nitrite and nitrate levels in the
Thyroxine preparation culture tank. All the parameters were
As for thyroxine (T4) experiment, 3 measured once a week, prior to 10 %
dosages used were 0, 0.5 and 1 ppm. water change.
[ DOI: 10.18331/SFS2021.8.1.7 ]
Hormone immersions were carried out
for 30 minutes at the initiation of Feeding
experiment on fry of 28 DAH. Fry were All the broodstocks and the fry were fed
stocked into polystyrene box measuring to observed satiation twice daily at 0800
34x21x19cm. L-thyroxine or T4 powder and 1700. Grindal worm, EnchytraeusJournal of Survey in Fisheries Sciences 8(1) 2021 93
buchholzi, Artemia nauplii (INVE quality sampled from the natural habitat
Aquaculture Inc., Belgium) and showed that this species can thrive well
sometimes Daphnia magna neonates in pH 4.3 to 5.9 (Table 1). Thus the pH
were used as food for the broodstock. tested in this experiment was 4.5, 5.0,
While, green water and microworm, and 5.5, and 6.0.
Panagrellus redivivus were fed to the fry
from the age of 3 DAH. Except for Table 1: Water quality of the natural habitat
of Parosphromenus tweediei at Sri
feeding regime experiment, fry and Bunian, Pontian, Johor.
juveniles were given mixture of Artemia Water parameters Range
nauplii and grindal worm, Enchytraeus pH 4.3 - 5.9
Temperature (˚C) 24.4 - 26.7
buchholzi. Uneaten food and faecal
Dissolved oxygen, DO 6.52 - 8.04
matters were removed by siphoning. All (mg/L)
the live foods were produced in the
laboratory. Significant differences (p94 Zulkifile et al., Effects of pH, feeding regime and thyroxine on growth and survival of …
Table 2: Mean of total length (TL) and survival of Parosphromenus tweediei fry at different pH
during the 56 days experimental period.
Day after Parameter pH
hatch (DAH) 4.5 5.0 5.5 6.0
7 (Initial) TL (mm) 4.71 ± 0.08a 4.71 ± 0.09a 4.71 ± 0.05a 4.71 ± 0.04a
14 TL (mm) 5.52 ± 0.06b 5.46 ± 0.03b 5.24 ± 0.03a 5.23 ± 0.03a
Survival (%) 100 ± 0.00a 100 ± 0.00a 94.44 ± 0.23a 91.67 ± 0.28a
21 TL (mm) 6.76 ± 0.04c 6.50 ± 0.04b 6.22 ± 0.03a 6.16 ± 0.02a
Survival (%) 100 ± 0.00b 100 ± 0.00b 88.89 ± 0.32ab 80.56 ± 0.40a
28 TL (mm) 8.02 ± 0.01d 7.73 ± 0.03c 7.27 ± 0.06b 7.11 ± 0.05a
Survival (%) 100 ± 0.00b 100 ± 0.00b 83.33 ± 0.38ab 77.78 ± 0.42a
35 TL (mm) 9.55 ± 0.05d 9.15 ± 0.06c 8.33 ± 0.09b 8.10 ± 0.06a
Survival (%) 100 ± 0.00b 100 ± 0.00b 77.78 ± 0.42a 75.00 ± 0.44a
42 TL (mm) 11.49 ± 0.01d 10.86 ± 0.03c 9.52 ± 0.03b 9.32 ± 0.04a
Survival (%) 100 ± 0.00b 100 ± 0.00b 77.78 ± 0.42a 77.78 ± 0.42a
49 TL (mm) 13.64 ± 0.06d 12.95 ± 0.06c 11.10 ± 0.04b 10.93 ± 0.04a
Survival (%) 100 ± 0.00b 100 ± 0.00b 77.78 ± 0.42a 77.78 ± 0.42a
56 TL (mm) 16.12 ± 0.02d 15.25 ± 0.03c 13.10 ± 0.01b 12.72 ± 0.01a
Survival (%) 100 ± 0.00b 100 ± 0.00b 77.78 ± 0.42a 77.78 ± 0.42a
63 TL (mm) 18.93 ± 0.01d 17.85 ± 0.01c 15.22 ± 0.02b 14.70 ± 0.02a
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b b a
Survival (%) 100 ± 0.00 100 ± 0.00 77.78 ± 0.42 77.78 ± 0.42a
Mean values with the same superscripts within the same row are not significantly different (p>0.05); ± SD
There were no significant differences in pH 4.5 and 5.0 as compared to fry in
(p>0.05) in survival of 7 – 14 DAH fry. pH 5.5 and 6.0. These findings suggested
At 21 DAH, fry survival was highest in that P. tweediei fry is most suitable in
pH 4.5 followed by pH 5.0, and the acidic environment evident with the
lowest (p0.05) when compared to pH 4.5, 5.0 pH 4.5, 5.0, 5.5 and 6.0. However
and 5.5. At 35 DAH, fry survival was ammonia and nitrite were significantly
higher (pJournal of Survey in Fisheries Sciences 8(1) 2021 95
While ranges for ammonia, nitrite and
nitrate were 0.25 -1.00 ppm, 0 1 ppm and
0 – 5 ppm, respectively.
Table 3: Water quality parameters in culture tank at different pH throughout the 56 days
experimental period
Water Parameter pH
4.5 5.0 5.5 6.0
Dissolved oxygen (mg/L) 3.88 ± 0.10a 3.69 ± 0.02a 3.85 ± 0.03a 3.84 ± 0.11a
Range 3.50 - 4.50 3.05 - 3.90 3.82 - 4.15 3.75 - 4.65
Temperature (°C) 25.45 ± 0.11a 25.31 ± 0.43a 25.58 ± 0.56a 25.71 ± 0.31a
Range 25.5 - 27.0 25.5 - 27.0 25.0 - 27.0 25.0 - 27.0
Ammonia (ppm) 0.65 ± 0.07b 0.59 ± 0.08b 0.42 ± 0.03a 0.27 ±0.03a
Range 0.50 – 1.00 0.25 - 1.00 0.25 - 0.50 0.25 - 0.50
Nitrite (ppm) 0.35 ± 0.03b 0.38 ± 0.00b 0.17 ± 0.02a 0.19 ± 0.03a
Range 0.00 - 1.00 0.00 - 0.50 0.00 - 0.25 0.00 - 0.50
Nitrate (ppm) 3.54 ± 0.36c 2.50 ± 0.00b 2.50 ± 0.00b 1.67 ± 0.36a
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Range 0.00 - 5.00 0.00 - 5.00 0.00 - 5.00 0.00 - 5.00
Mean values with the same superscripts within the same row are not significantly different (p>0.05); ± SD.
Feeding regime treatments. But at 28 DAH, AMG2
Results from the feeding regime showed significantly higher (p96 Zulkifile et al., Effects of pH, feeding regime and thyroxine on growth and survival of …
Table 4: Mean of total length (TL) and survival of Parosphromenus tweediei fry with different feeding
regime for 56 days experimental period.
Day after Parameter Feeding Regime
hatch (DAH) AM AMG1 AMG2
7 (Initial) TL (mm) 4.92 ± 0.03a 4.92 ± 0.03a 4.92 ± 0.05a
14 TL (mm) 5.70 ± 0.03a 5.71 ± 0.07a 5.72 ± 0.08a
a a
Survival (%) 100.00 ± 0.00 96.67 ± 0.18 96.67 ± 0.18a
21 TL (mm) 6.90 ± 0.02a 6.91 ± 0.03a 6.91 ± 0.02a
a a
Survival (%) 100.00 ± 0.00 96.67 ± 0.18 96.67 ± 0.18a
28 TL (mm) 8.02± 0.03a 8.01 ± 0.01a 8.22 ± 0.02b
Survival (%) 96.67 ± 0.18a 93.33 ± 0.25a 96.67 ± 0.18a
a a
35 TL (mm) 9.33 ± 0.06 9.30 ± 0.04 9.61 ± 0.02b
Survival (%) 96.67 ± 0.18a 93.33 ± 0.25a 96.67 ± 0.18a
a a
42 TL (mm) 11.15 ± 0.02 11.16 ± 0.02 11.60 ± 0.02b
a a
Survival (%) 96.67 ± 0.18 93.33 ± 0.25 96.67 ± 0.18a
49 TL (mm) 13.23 ± 0.02a 13.25 ± 0.02a 13.82 ± 0.02b
a a
Survival (%) 96.67 ± 0.18 93.33 ± 0.25 96.67 ± 0.18a
56 TL (mm) 15.32 ± 0.04a 15.64 ± 0.03b 16.26 ± 0.05c
a a
Survival (%) 96.67 ± 0.18 93.33 ± 0.25 96.67 ± 0.18a
63 TL (mm) 17.97 ± 0.02a 18.24 ± 0.01b 18.91 ± 0.02c
a a
Survival (%) 96.67 ± 0.18 93.33 ± 0.25 96.67 ± 0.18a
Mean values with the same superscripts within the same row are not significantly different (p>0.05); ±SD.
AM: Artemia nauplii (DAH6-20) - Moina sp. (DAH21-63)
AMG1: Artemia nauplii (DAH6-20) - Moina sp. (DAH21-43) - Grindal worm (DAH44-63)
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AMG2: Artemia nauplii (DAH6-20) - Moina sp. (DAH27-48) - Grindal worm (DAH49-63)
Table 5: Water quality parameters in culture tank at different feeding regimes throughout the 56 days
experimental period
Feeding regime
Water Parameter
AM AMG1 AMG2
Dissolved oxygen (mg/L) 3.91 ± 0.00a 3.91 ± 0.02a 3.91 ± 0.01a
Range 3.75 - 4.65 3.75 - 4.65 3.75 - 4.65
Temperature (°C) 25.36 ± 0.04a 25.36 ± 0.15a 25.40 ± 0.12a
Range 25.0 - 26.0 25.0 - 26.0 25.0 - 26.0
a a
Ammonia (ppm) 0.71 ± 0.01 0.71 ± 0.01 0.71 ± 0.01a
Range 0.25 - 1.00 0.25 - 1.00 0.25 - 1.00
Nitrite (ppm) 0.45 ± 0.05a 0.44 ± 0.04a 0.40 ± 0.01a
Range 0.00 - 0.50 0.00 - 0.50 0.00 - 0.50
a a
Nitrate (ppm) 4.76 ± 0.04 4.76 ± 0.05 4.72 ± 0.01a
Range 0.00 – 5.00 0.00 – 5.00 0.00 – 5.00
Mean values with the same superscripts within the same row are not significantly different (p>0.05); ±SD.
[ DOI: 10.18331/SFS2021.8.1.7 ]
Water pH in all tanks were maintained at 4.5 throughout the experimental period
AM: Artemia nauplii (DAH6-20) - Moina sp. (DAH21-63)
AMG1: Artemia nauplii (DAH6-20) - Moina sp. (DAH21-43) - Grindal worm (DAH44-63)
AMG2: Artemia nauplii (DAH6-26) - Moina sp. (DAH27-48) - Grindal worm (DAH49-63)Journal of Survey in Fisheries Sciences 8(1) 2021 97
Mean values for DO, temperature, fry was 7.9mm. At 56 DAH, highest
ammonia, nitrite, nitrate were not (p0.05) between immersion with 0.5 ppm of T4, followed
all feeding regimes (Table 5).
by 1.0ppm T4, and the lowest (p0.05) was observed for fry
feeding regimes did not affect the water
immersed among the three dosages of T4
quality in the experimental tanks.
at 56 and 84 DAH. These results showed
T4 at 0.5 ppm improved the growth, and
Thyroxine
most importantly no adverse effect on
Fry of P. tweediei were immersed with
survival of P. tweediei fry.
thyroxine (T4) at dosages of 0, 0.5, 1.0
ppm (Table 6). Initial total length TL of
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Table 6: Mean of total length (TL) and survival of Parosphromenus tweediei fry for 56 days
experimental period after different dosages of thyroxine immersion
Day after Thyroxine (ppm)
Parameters
hatch (DAH) 0 0.5 1.0
28 Mean initial TL (mm) 7.90 ± 0.02a 7.90 ± 0.03a 7.90 ± 0.05a
Mean TL (mm) 20.60 ± 0.08a 21.49 ± 0.05c 21.01 ± 0.05b
56 Survival (%) 100.00 ± 0.00a 90.0 ± 0.31a 90.0 ± 0.31a
Mean TL (mm) 29.73 ± 0.08a 30.49 ± 0.10c 29.99 ± 0.06b
84 a a
Survival (%) 100.00 ± 0.00 90.0 ± 0.31 90.0 ± 0.31a
Mean values with the same superscripts within the same row are not significantly different (p>0.05); ±SD.
Mean values for DO, temperature, 0– 0.5 ppm, 0 – 0.25 ppm and 0 – 5.00
ammonia, nitrite, nitrate were not ppm respectively. These water quality
significantly different (p>0.05) between results showed no significant different
0, 0.5 and 1.0 ppm of T4 treatments (p98 Zulkifile et al., Effects of pH, feeding regime and thyroxine on growth and survival of …
Table 7: Water quality parameters in culture tank of P. tweediei fry with different thyroxine dosage
throughout the 56 days experimental period
Thyroxine (ppm)
Water Parameter
0 0.5 1.0
Dissolved oxygen (mg/L) 3.89 ± 0.02a 4.10 ± 0.01a 3.91 ± 0.03a
Range 3.85 - 4.18 3.98 - 4.15 3.86 - 4.11
Temperature (°C) 25.53 ± 0.04a 25.61 ± 0.02a 25.54 ± 0.05a
Range 25.4 - 26.3 25.3 - 26.0 25.4 - 26.3
Ammonia (ppm) 0.41 ± 0.03a 0.45 ± 0.01a 0.41 ± 0.02a
Range 0.00 - 0.50 0.00 - 0.50 0.00 - 0.50
Nitrite (ppm) 0.20 ± 0.01a 0.24 ± 0.02a 0.20 ± 0.01a
Range 0.00 - 0.25 0.00 - 0.25 0.00 - 0.25
Nitrate (ppm) 4.76 ± 0.04a 4.76 ± 0.05a 4.72 ± 0.01a
Range 0.00 – 5.00 0.00 – 5.00 0.00 – 5.00
Mean values with the same superscripts within the same row are not significantly different (p>0.05); ± SD.
Discussion According to Boyd (1982), growth
Hydrogen ion concentration is reflected and reproduction of aquatic species will
as pH in water. It plays an important role decrease as pH lowered to less than 6.5
for aquatic animals particularly fish for a prolonged period. In a gourami
(Miron et al., 2008). It affects the normal species, Trichogaster lalius, exposure to
physiological function of aquatic pH 5.7 will cause tremendous stress and
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organism, whereby the increase or even mortality (Sachin and Subhendu,
decrease will disrupts respiration, ion 2018). In the present study, findings from
exchange and ammonia excretion pH experiment clearly showed that not
(Schlenk and Benson, 2001; Mahassen et only P. tweediei are able to tolerate low
al., 2011). For freshwater fish, pH is a pH, but also grow and survive best in pH
major determinant for certain species 4.5 to 5.0. At pH 5.5 and 6.0, growth and
because the tolerance limit varies survival of P. tweediei are adversely
depending on species (West et al., 1997; affected, possibly due to the decreased of
Mills et al., 2000; Greig et al., 2010). acid-base balance in species with
Most inland freshwaters inhabited by fish preference towards acidic condition.
have water pH range of 6.0 to 9.0 (Ellis, Wilkie and wood (1991) observed that if
1937). Nevertheless, there are fishes the pH is beyond tolerable range, fish
living in water with pH of 4.0 and 10.0 will die due to the inhibition of ammonia
(ORVWSC, 1955). In the present study, excretion through the gills. Nonetheless,
P. tweediei broodstock were sampled pH tolerance in fish will also depend on
from natural habitat at Pontian, Johor, various other factors such temperature
[ DOI: 10.18331/SFS2021.8.1.7 ]
with pH range of 4.3 to 5.9. This result and DO (Mckee and Wolf, 1963).
showed that this fish can tolerate acidic In addition to the limited availability of
water condition. Even Kottelat and Ng live food, prey preferences further
(2005) reported P. tweediei lives in water aggravate the condition which may
with pH 4.0. affects growth and survival of fish fryJournal of Survey in Fisheries Sciences 8(1) 2021 99
(Pham, 2001). It is quite common that abundance, soft body, motile and short
type and size of prey selected by fry life cycle (Luna-Figueroa et al., 2000;
change during larval developmental Erdogan and Olmez, 2009). As live food,
stage. In most cases, fish fry generally this worm is able to contribute 11.6% of
preferred small size prey (Srikrishan et protein (Bouguenec, 1992). The effect
al., 2017). The abundance of small on growth of Pterophyllum scalare fry
zooplankton influences feeding success fed with grindal worm is equivalent to
(Hansen and Wahl, 1981). However, that of artificial feed (Luna-Figueroa et
bigger prey is preferred as fry grew larger al., 2000) but lesser than Moina,
and the size of mouth becomes wider decapsulated Artemia and Daphnia
(Gill and Hart, 1996). Therefore, Laron (Sorianao and Hernendez, 2002, Ortega-
(2001) suggested the combination of Salas et al., 2009; Santillan, 2011;
small and large live food to provide good Jimenez-Rojas et al., 2012). This is
results. In the present study, feeding basically due to other live foods having
regime AMG2 utilizes Artemia nauplii comparatively higher protein content,
for fry from 6 to 20 DAH, then Moina sp. better distribution and longer availability
for fry from 27 to 48 DAH, and finally in the water column, and at the same time
grindal worm for fry from 49 to 63 DAH, these characteristics reduce the creation
produced the best growth of fry from 28 of dominant fish during feeding
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to 63 DAH. Artemia nauplii were given (Sorianao and Hernendez, 2002; Luna-
for a longer time in AMG2. According to Figueroa et al., 2010; Takahashi et al.,
Stickney (1994), Artemia nauplii is a 2010). Mosquito larvae and blood worm
must include food in general feeding which are larger than grindal worm can
strategy for fish fry. Srikrishan et al. also be considered for P. tweediei fry
(2017) reported that feeding with from 44 DAH onwards.
Artemia nauplii resulted in higher Various studies have shown the
survival as compared to Moina sp. growth promoting effect of thyroxine
Consequently, the longer used of (T4) to fish larvae (Nacario, 1983; Lam
Artemia nauplii as initial food, the higher and Sharma, 1985; Reddy and Lam,
the survival rate of fry. 1992; Woodhead, 1996). In the present
Laron (2001) suggested the study, immersion of T4 at 0.5 and
combination of small and large live food 1.00ppm resulted in significantly
to improve growth and survival catfish accelerated growth of P. tweediei fry.
fry. In the present study, the inclusion of The half-life of T4 is 5-7 days, where it
grindal worms, Enchytraeus buchholzi in will convert into Tri-iodothyronine (T3)
[ DOI: 10.18331/SFS2021.8.1.7 ]
AMG1 and AMG2 resulted in better or eliminated from the system (Barac-
growth of P. tweediei fry as compared to Latac, 2009). Study by Nacario (1983)
without it in AM. Grindal worm is being showed that lower dosage T4 increases
used in aquaculture due to their the length of the pectoral fin
nutritional characteristics, availability, significantly, while at higher dosage it100 Zulkifile et al., Effects of pH, feeding regime and thyroxine on growth and survival of …
resulted in the pectoral fin malformation this low pH condition. Nitrate highest
and scoliosis. Overall, the growth level in this study was at 5ppm, however,
performance of fish may differ depends this level is considered as non-toxic to
on its sensitivity at different stages (Lam fish (Francis-Floyd et al., 2009).
et al., 1985). In the present study, Based on the findings of this study, P.
scoliosis was observed on some of the P. tweediei fry can be successfully cultured
tweediei fry immersed with 1 ppm T4. in water with pH 4.5 to 6.0. The
Hence, it is highly recommended to use observation on early developmental
the lower dosage of 0.5ppm T4 stages showed that this species can reach
immersion for P. tweediei fry. total length of 18.93 mm at 63 DAH.
In the present study, P. tweediei fry Parosphromenus tweediei is able to
are able to tolerate DO as low as tolerate acidic water condition and higher
3.05ppm, while temperature as low as 25 ammonia levels, thus a considerably
°C. As for ammonia, nitrite and nitrate, hardy fish. During fry to juvenile stage,
P. tweediei fry are able tolerate as high as it can be fed using combination of live
1, 1 and 5 ppm, respectively. Ammonia food consisting Artemia nauplii, Moina
level is influenced by temperature and sp. and grindal worm. Application of
pH. As water temperature rises ammonia thyroxine (T4) at 0.5 ppm per one time
become more toxic (Levit, 2010). immersion at early stage can
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However, the temperatures recorded in significantly promote the growth of P.
all the experiments in the present study tweediei fry. Cost of production can be
were quite low, ranged from 25 to 27°C. reduced with the application of T4 as the
As for pH, growth of nitrifying bacteria hormone will accelerate the growth of fry
is inhibited at low pH, thus the slow thus shorten the culture period.
conversion of the ammonia to nitrite and
nitrates. Nitrification process is inhibited Acknowledgements
at pH below 5.8 (Princic et al., 1998). As Many thanks to Zahar Zakaria, Ilham
consequences, ammonia will be Norhakim, and Akiran Hassan for ideas
accumulated in the water and to certain for the project and helping with
level become toxic to fish. In addition, sampling. The authors also would like to
the accumulation of nitrite or nitric oxide express their gratitude to the Institute of
can occur when DO level is below 2 Bioscience and the Department of
mg/L (Goreau et. al., 1980; Painter, Aquaculture, Faculty of Agriculture,
1986). However, at low pH, ionized Universiti Putra Malaysia for providing
ammonia (NH4+) become less toxic as the materials and facilities for this study.
[ DOI: 10.18331/SFS2021.8.1.7 ]
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