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International Journal of Trend in Scientific Research and Development (IJTSRD)
Volume: 3 | Issue: 4 | May-Jun 2019 Available Online: www.ijtsrd.com e-ISSN: 2456 - 6470
@ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 417
Oral Administration of Pulverized Wood Charcoal
on Growth, Feed Utilization, Survival and Waste
Excretion of Red Tilapia (Oreochromis Sp)
Jaypee. S. Samson
Faculty Researcher, College of Fisheries, Freshwater Aquaculture Centre,
Central Luzon State University, Science City of Muñoz, Nueva Ecija, Philippines
How to cite this paper: Jaypee. S.
Samson "Oral Administration of
Pulverized Wood Charcoal on Growth,
Feed Utilization, Survival and Waste
Excretion of Red Tilapia (Oreochromis
Sp)" Published in International Journal
of Trend in Scientific Research and
Development
(ijtsrd), ISSN: 2456-
6470, Volume-3 |
Issue-4, June 2019,
pp.417-421, URL:
https://www.ijtsrd.c
om/papers/ijtsrd23
724.pdf
Copyright © 2019 by author(s) and
International Journal of Trend in
Scientific Research and Development
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Attribution License (CC BY 4.0)
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by/4.0)
ABSTRACT
A 35-day feeding experiment was conducted to evaluatetheeffectsofpulverized
wood charcoal (PWC) on the growth, feed utilization, survival and waste
excretion of red tilapia (Oreochromis sp.). Ninety red tilapias with an initial
weight of 7.32 ± 1.31 g were acclimatized and randomly introduced into 9
aquaria in 3 groups with 10 fish per aquarium. Different levels of PWC (0 (T0),
1% (T1), and 2% (T2)) were incorporated in the diets of red tilapia for four
weeks. After the experiment, average body weight (ABW), specific growth rate
(SGR), relative growth rate (RGR) weight gain (WG), feed conversionratio(FCR)
and survival rate (SR) of the experimental fish were computed. No significant
differences were observed on feed utilization, survival, and waste excretion of
the fish. Furthermore, ABW and WG of the experimental fish did not show any
significant difference, however, significantlyhigher(P <0.05) SGRand RGRwere
observed from T1 compared to the other treatments. Consequently, this study
showed that supplementation of 1% PWC in the diet is the most suitable in
improving the growth performance of red tilapia.
Keywords: Ammonia, Aquaculture, Feed additive, Growth promotion, Red tilapia
INTRODUCTION
Tilapia species are known for their ability to tolerate a wide range of
environmental factors and stressful conditions. They areconsidered asoneof the
most important fish species since the early twentieth century till present
(Fitzsimmons 2000). Red tilapia species became popular among culturistsdueto
the similarity in shape with sea bream and excellent growth and feed conversion
rates among fresh or brackish water species (Liao & Chen 1983).
FAO (2010) reported that in 2009 the global production of
red tilapia was 38 064 ton. Fish require variable protein
levels in their feeds to maintain optimal growth andsurvival
that is why the quality of feed is one crucial factor in the
aquaculture industry. However, the high levels of protein in
fish feeds loads more wastes to the aquatic environment.
Common wastes coming from aquaculture facilities are the
nitrogenous waste, which is toxic to aquatic organisms and
affects their growth and survival. One source of these
nitrogenous wastes such as ammonia (Kikuchi etal. 1995) is
the undigested commercial feeds given to the cultured
species (Quiayum et al. 2014). Aside from that, fish and
aquatic organisms contribute to the increase of ammonia
level in the water through their wasteexcretions(Floydetal.
1990; Yu et al. 2007). As part of the growth and survival of
animals, they continuously excrete wastesas aresult of their
metabolic process in acquiring energy and nutrients from
their food. Ammonia is the principal metabolic waste
excreted by the fish so accumulateseasilyinaquaticsystems.
Because of its negative impact on the health of the fish,
ammonia should be regulated before it accumulates and
harms the fish. (Floyd et al 1990).
Ammonia is considered a very toxic factor to fish that limits
their survival and growth (Person-LeRuyet etal. 1995). High
levels of ammonia may result to gill damage, reduced
growth, poor feed conversion, and lower resistance to
diseases, increased breathing, cardiac output and oxygen
uptake and may result to convulsions and death (Floyd et al.
1990, Buttner et al. 1993, Yu et al. 2007, Sichula et al. 2011,
Bhatnagar & Devi, 2013). Ammonia in water is composed of
two forms; the ionized (NH4
+) an un-ionized (NH3)ammonia
(Floyd et al. 1990, Yu et al. 2007). Between these two forms
of ammonia, the un-ionized form is more toxic than the
ionized form because of its ability to move across cell
membranes (El-Sherif & El-Feky 2008; Yu et al. 2007,
Bhatnagar & Devi 2013). The predominantformof ammonia
present in water is affected by temperature and pH of the
aquatic system (Floyd et al. 1990). Both forms are termed as
"total ammonia nitrogen (TAN)” (Floyd et al. 1990, Yu et al.
2007).
Modifying the feeds given to the cultured species is one of
the recommended ways to reduce the nitrogenous waste
from aquaculture. Currently, a natural substance such as
charcoal is used in formulating animal feeds becauseoftheir
IJTSRD23724
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 418
ability to absorb nitrogen and ammonia and helps in
activating the intestinal function of animals (Quiayum et al.
2014). Charcoal is a form of carbon, which is generally
produced as a carbonaceous residue of wood and other
industrial wastes after heating the organic matter. It
contains 70 to 90% pure carbon and several minerals such
as calcium, potassium, iron, sodium, copper, zinc,
manganese, magnesium, etc. (Brouwer et al.1996).Charcoal
has a large surface area giving it countless bonding sites
which can be used in adsorbing toxins, gases, and drugs
without any specific action (Osol, 1975). It has been used as
a feed additive for many terrestrial animals because of its
ability to absorb gases especially nitrogen and ammonia,
activate the intestinal function, and eliminate the poisons
and impurities fromthegastrointestinaltractof land animals
(Banner et al. 2000, Bisson et al. 2001, Kutlu et al. 2001,
Samanya and Yamauchi 2001, Mekbungwan et al. 2004). It
has also been proven to be a good feed additive in the feeds
of aquatic animals such as tiger pufferfish (Takifugu
rubripes) Japanese flounder (Paralichthys olivaceus),
Pangasius hypophthalmus, Clarias gariepinus, Nile tilapia
(Oreochromis niloticus), red tilapia hybrid (Oreochromis
mossambicus × Oreochromis niloticus) and Gilthead
Seabream (Sparus aurata) (Thu et al. 2009, 2010, Quaiyum
et al. 2014, Jahan et al. 2014, Lawal et al, 2018, Nopadon et
al. 2015, Michael et al. 2015, Michael & Helal 2018).
This study aimed to evaluate the effect of pulverized wood
charcoal on the on growth, feed utilization, survival and
waste excretion of Red tilapia (Oreochromis sp.)
MATERIALS AND METHODS
Experimental fish and feeds: Red tilapia fingerlings were
obtained from Freshwater Aquaculture Centre, Central
Luzon State University, Science City of Muñoz, Nueva Ecija,
Philippines. Experimental fish with an initialweightof7.32±
1.31 g were acclimatized for one week and were starved for
24 hours before the commencement of the experiment. Ten
randomly selected red tilapia were placed in each glass
aquarium and fed the with their designated feed thriceaday
(8:00, 12:00 and 16:00), with an initial feeding rate of 10%
of their body weight. Sampling was done once every two
weeks to measure their weight and adjust the feeding rate.
This study was designed randomly comprising of 3
treatments with each treatment replicated thrice.
Commercially-available diet (35% protein, 4%fat) was used
as the control for the experiment. Commercial diet was
added with 0% (Control) (T0), 1% (T1) and 2% (T2) PWD
and water (200 mL kg-1) (Cho and Lee, 2012). Mixtureswere
mixed thoroughly and dried at room temperature before
storing at -20 °C.
Growth performance and Feed Utilization: To assess the
growth performance and feed utilizationof theexperimental
fish, the following data weregathered duringtheexperiment
which includes specific growth rate (SGR), relative growth
rate (RGR), weight gain (WG), survival rate (SR) and feed
conversion ratio (FCR). Parameters were calculated as
follows:
Water Quality Monitoring: Water quality parameterssuch
as temperature, dissolved oxygen (DO) and pH were
measured using DO meter and pH meter. For the analysis of
TAN, water samples were collected fromtheaquariumtanks
and were placed in polyethylene (PE) bottles prior to the
analysis in the laboratory. Analysis of TAN was done every
week before water change following the phenate method.
Briefly, 25 to 50 mL of water sample was filtered through
Whatman No. 42, and 10 mL of the filtered sample was
transferred into a 50 mL beaker in a magnetic stirrer. While
stirring, 1 drop of manganese sulfate(MnSO4•H2O)solution,
0.5 mL oxidizing solution and 0.6 mL phenate solution was
added. Then, removed from stirrer and allowed 15 minutes
for maximumcolor developmentbeforetransferringtransfer
to the 1-cm cuvette. Ammonia-free distilled water (10.00
mL) (reagent blank) and 10.00 mL of the 0.300 mg · L-1
solution of total ammonia-nitrogen (standard) were carried
through the procedure with each set. With the
spectrophotometer at 630 nm, set 0.0 absorbance (100%
transmittance) with the reagent blank, the absorbances of
the standard and the samples were read. Lastly, the
concentrations of the TAN were calculated using the
formula:
Statistical Analysis: Results for the growth, survival and
feed utilization were subjected to one-way analysis of
variance (ANOVA) followed by Duncan’s test at a significant
level of P<0.05.
RESULTS
Table1. Growth performance and feed utilization of Red tilapia fed on diets supplemented with different levels of
pulverized wood charcoal (PWC) for 35 days.
Parameters T0 (0% PWC) T1 (1% PWC) T2 (2% PWC)
Initial ABW, g 7.80 ± 0.85a 6.85 ± 2.00a 7.32 ± 1.23a
Final ABW, g 14.71 ± 2.72a 15.18 ± 1.45a 13.84 ± 1.85a
SGR (%) 0.78 ± 0.14b 1.22 ± 0.10a 0.80 ±0.17b
RGR (% · day-1) 87.91 ± 21.79b 168.72 ± 21.99a 91.25 ± 24.52b
WG, g 6.91 ± 2.11a 8.33 ± 2.34a 6.52 ± 1.57a
FCR 3.72 ± 0.36a 2.45 ± 0.98a 2.98 ± 0.45a
SR (%) 36.67 ± 5.77a 46.67 ± 15.27a 53.33 ± 37.85a
PWC, pulverized wood charcoal; ABW, average body weight; SGR, specific growth rate; RGR, relative growth rate; WG, weight
gain; FCR, feed conversion ratio; SR, survival rate.
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 419
Means in column with the same letter are not significantly different at P < 0.05
Figure.1. Total ammonia nitrogen (TAN) co+ncentration of water of aquaria stocked by red tilapia fed on diets
supplemented with different levels of pulverized wood charcoal (PWC) for 35 days.
Results of growth performance, feed utilization,andsurvival,
at the end of the experiment is shown in Table 1. There were
no significant differences observed in the initial and final
ABW of the experimental fish. However, PWC inclusion of
1% (T1) significantly (P < 0.05) improved the SGR and RGR
of the experimental fish whencomparedtothecontrolgroup
(T0). WG from T1 was also numerically higher compared to
the other treatments (T0 and T2), however, the observed
increases were not statistically significant based on
comparisons with the other groups. Furthermore, although
FCR and SR were better on PWC-supplemented diets, no
significant differences were observed upon comparing with
the control group.
Water quality parameters such astemperature(28.75± 0.85
ºC), pH (8.4 ± 0.22), and DO (2.5 ± 0.83 mg · L-1) did notshow
any significant differences among the treatments. Levels of
TAN concentration were found to be varying throughoutthe
culturing period. During the 1st week until the 3rd week of
feeding experiment, a high concentration of TAN was
recorded in PWC-supplementeddiets,withreadingsof0.996
to 1.016 mg · L-1, respectively. As the experiment progress,
the level of TAN decreases (Fig 1). In the 4th and 5th week of
the feeding trial, TAN levels from PWC-supplemented diets
were numerically lower, however, the observed decrease
was not statistically significant based on comparisons with
the control group.
DISCUSSION
The quality of the fish diet is one of the most important
factors to consider when it comes to the optimal growth
potential of an aquaculture organism. Over the years,
numerous efforts were made to formulate a diet that can
provide optimal growth, and made of readily available and
inexpensive ingredients to reduce the cost of feed
production. However, despite the capabilities of the feeds to
sustain good fish growth rates, they have a negative
environmental impact due to their high nitrogenous wastes
particularly ammonia that constitutes almost 80% of these
wastes (Kikuchi et al. 1995). With that, the use of feed
additives especially charcoals to reduce the nitrogenous
waste coming from the aquaculture industry and promoting
better growth to the cultured species is gaining popularity.
The inclusion of charcoal in the diets of animals hasreceived
attention due to its non-nutritive and nondigestible
characteristics giving it the ability to adsorb toxins from the
gastrointestinal tract thus preventing their absorption into
the body. Several researches have demonstrated that
charcoal-supplemented diets can improve fish growth, flesh
quality, and intestinal function (Boonanuntanasarn et al.
2014, Quiayum et al. 2014, Jahan et al. 2014, Moe et al. 2010,
Thu et al. 2009, 2010, Lawal et al. 2018, Nopadon et al.2015,
Michael & Helal 2018).
The present study aimed to evaluate the effect of different
levels of PWC in the feeds of red tilapia on their growth, feed
utilization, survival, and waste excretion. Results showed
that 1% inclusion of PWC in the diet significantly improved
the SGR and RGR of the fish. The result obtained from this
study is in parallel with the report of Nopadon et al. (2015),
wherein 1-3% of AC inclusion improved the growth
performance of Nile tilapia with optimal inclusion of 2%.
Also, Abdel-Tawwab et al. (2017) observed improvement in
the growth parameters of Nile tilapia at AC inclusion of 0.2-
2%, with optimal inclusion level of 0.7%. Additionally, the
inclusion of commercial wood charcoal(CWC)at3-4%inthe
diet of red tilapia hybrid (Oreochromis mossambicus ×
Oreochromis niloticus) improved fish survival, growth, and
feed utilization. In contrast, the present study found no
significant improvement in the growth of red tilapia at a 2%
level of PWC inclusion. Supplementation of charcoal in the
diets have also shown improved growth performance of
other aquatic animals. Thu et al. (2009) and Michael & Helal
(2018) reported that 4% of charcoal inclusion is the optimal
level for tiger puffer fish and gilthead seabream. While 0.5%
level of bamboo charcoal (BC) inclusion is reported by Moe
et al. (2010) to be the best in juvenile Japanese flounder. In
addition, Lawal et al. (2018) found significant improvement
in the growth of Clarias gariepinus with 0.25% level of AC
inclusion. Furthermore, Quiayum et al. (2014) and Jahan et
al. (2014) observed significant effects of BC-supplemented
diets in pangasiid catfish growth and ammonia loading,
however, they reported a dissimilar level of optimum
charcoal inclusion of 2% and 1%, respectively. On the other
hand, Boonanuntanasarn et al. (2014), reported that growth
parameters of Nile tilapia (O. niloticus) reared on diets
containing 10 g · kg-1 body weight AC only showed a
numerical improvement with no statistical significance.The
same result was observed by Mabe et al. (2017) in juvenile
common carp (Cyprinus carpio L.) fed with differentlevels of
BC in the diet. These results suggest that the optimal level of
charcoal inclusion in the diet of aquaticanimalsdiffer among
species, animal’s life stage, and the nature of charcoal.
The enhanced growth parameters in the present study may
be explained due to the capability of PWC to adsorb and
eliminate unwanted gases and toxic substances, which can
result to improved digestion, metabolism, and better
nutrient utilization and absorption (Kutlu et al. 2001,
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 420
Samanya & Yamauchi 2001, Jahan et al. 2014). Nevertheless,
since charcoal is a nonspecific adsorbent, its application in
the diets of animals may possibly causetheadsorptionof not
only toxins but also nutrients from the gastrointestinaltract.
Thus, positive and negative impacts on physiological
functions, growth and overall health of an organism can be
both expected (Boonanuntanasarn et al.2014,Nopadonetal.
2015).
The SR obtained in all treatments is noticeably low.
Mortalities of the experimental fish started during the 3rd
and 4th week of the experiment. As an observation, some
experimental fish are gliding theirbodies alongthesidesand
bottom of the aquarium. It is possible that during the
experiment, the fish acquired bacterial disease which is one
possible reason why the SR in all treatments is low. Aside
from that, the level of oxygen throughout the experiment
(2.5 ± 0.83 mg · L-1) is lower than the desired dissolved
oxygen level of at least 5 mg · L-1.
In the present study, no significant improvement was
observed in the waste excretion of the red tilapia. In
contrast, Thu et al. (2009) and (2010) reported that (BC)
supplementation reduced the ammonia-N values with tiger
puffer fish and Japanese flounder, respectively.Michaeletal.
(2015) also observed a gradual decrease in ammonia levels
in the rearing water area as the CWC levels in red tilapia
feeds increases. The same observation was noted by
Quiayum et al. (2014), where they found out that increasing
level of charcoal incorporation in fish feeds results to lower
ammonia level in the water. The possible reason with this is
that the duration of the experiment (35 days) is not enough
to observe the effect of PWC in the waste excretion of the
fish.
CONCLUSION
In conclusion, the result of the present study showed that
1% inclusion of PWC in the diet improved the growth
performance of the red tilapia (Oreochromis sp.). Therefore,
PWC at 1% can be a useful feed additive in the diet of red
tilapia as it is cheap and readily available.
ACKNOWLEDGEMENT
The author would like to acknowledge the assistance of the
College of Fisheries and Freshwater Aquaculture Centre
where the study was conducted. Also, the author would like
to extend his appreciation to the Department of Science and
Technology–Accelerated Science and Technology Human
Resource Development Program (DOST-ASTHRDP) for
financial assistance during the experiment.
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Oral Administration of Pulverized Wood Charcoal on Growth, Feed Utilization, Survival and Waste Excretion of Red Tilapia Oreochromis Sp

  • 1. International Journal of Trend in Scientific Research and Development (IJTSRD) Volume: 3 | Issue: 4 | May-Jun 2019 Available Online: www.ijtsrd.com e-ISSN: 2456 - 6470 @ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 417 Oral Administration of Pulverized Wood Charcoal on Growth, Feed Utilization, Survival and Waste Excretion of Red Tilapia (Oreochromis Sp) Jaypee. S. Samson Faculty Researcher, College of Fisheries, Freshwater Aquaculture Centre, Central Luzon State University, Science City of Muñoz, Nueva Ecija, Philippines How to cite this paper: Jaypee. S. Samson "Oral Administration of Pulverized Wood Charcoal on Growth, Feed Utilization, Survival and Waste Excretion of Red Tilapia (Oreochromis Sp)" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456- 6470, Volume-3 | Issue-4, June 2019, pp.417-421, URL: https://www.ijtsrd.c om/papers/ijtsrd23 724.pdf Copyright © 2019 by author(s) and International Journal of Trend in Scientific Research and Development Journal. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0) (http://creativecommons.org/licenses/ by/4.0) ABSTRACT A 35-day feeding experiment was conducted to evaluatetheeffectsofpulverized wood charcoal (PWC) on the growth, feed utilization, survival and waste excretion of red tilapia (Oreochromis sp.). Ninety red tilapias with an initial weight of 7.32 ± 1.31 g were acclimatized and randomly introduced into 9 aquaria in 3 groups with 10 fish per aquarium. Different levels of PWC (0 (T0), 1% (T1), and 2% (T2)) were incorporated in the diets of red tilapia for four weeks. After the experiment, average body weight (ABW), specific growth rate (SGR), relative growth rate (RGR) weight gain (WG), feed conversionratio(FCR) and survival rate (SR) of the experimental fish were computed. No significant differences were observed on feed utilization, survival, and waste excretion of the fish. Furthermore, ABW and WG of the experimental fish did not show any significant difference, however, significantlyhigher(P <0.05) SGRand RGRwere observed from T1 compared to the other treatments. Consequently, this study showed that supplementation of 1% PWC in the diet is the most suitable in improving the growth performance of red tilapia. Keywords: Ammonia, Aquaculture, Feed additive, Growth promotion, Red tilapia INTRODUCTION Tilapia species are known for their ability to tolerate a wide range of environmental factors and stressful conditions. They areconsidered asoneof the most important fish species since the early twentieth century till present (Fitzsimmons 2000). Red tilapia species became popular among culturistsdueto the similarity in shape with sea bream and excellent growth and feed conversion rates among fresh or brackish water species (Liao & Chen 1983). FAO (2010) reported that in 2009 the global production of red tilapia was 38 064 ton. Fish require variable protein levels in their feeds to maintain optimal growth andsurvival that is why the quality of feed is one crucial factor in the aquaculture industry. However, the high levels of protein in fish feeds loads more wastes to the aquatic environment. Common wastes coming from aquaculture facilities are the nitrogenous waste, which is toxic to aquatic organisms and affects their growth and survival. One source of these nitrogenous wastes such as ammonia (Kikuchi etal. 1995) is the undigested commercial feeds given to the cultured species (Quiayum et al. 2014). Aside from that, fish and aquatic organisms contribute to the increase of ammonia level in the water through their wasteexcretions(Floydetal. 1990; Yu et al. 2007). As part of the growth and survival of animals, they continuously excrete wastesas aresult of their metabolic process in acquiring energy and nutrients from their food. Ammonia is the principal metabolic waste excreted by the fish so accumulateseasilyinaquaticsystems. Because of its negative impact on the health of the fish, ammonia should be regulated before it accumulates and harms the fish. (Floyd et al 1990). Ammonia is considered a very toxic factor to fish that limits their survival and growth (Person-LeRuyet etal. 1995). High levels of ammonia may result to gill damage, reduced growth, poor feed conversion, and lower resistance to diseases, increased breathing, cardiac output and oxygen uptake and may result to convulsions and death (Floyd et al. 1990, Buttner et al. 1993, Yu et al. 2007, Sichula et al. 2011, Bhatnagar & Devi, 2013). Ammonia in water is composed of two forms; the ionized (NH4 +) an un-ionized (NH3)ammonia (Floyd et al. 1990, Yu et al. 2007). Between these two forms of ammonia, the un-ionized form is more toxic than the ionized form because of its ability to move across cell membranes (El-Sherif & El-Feky 2008; Yu et al. 2007, Bhatnagar & Devi 2013). The predominantformof ammonia present in water is affected by temperature and pH of the aquatic system (Floyd et al. 1990). Both forms are termed as "total ammonia nitrogen (TAN)” (Floyd et al. 1990, Yu et al. 2007). Modifying the feeds given to the cultured species is one of the recommended ways to reduce the nitrogenous waste from aquaculture. Currently, a natural substance such as charcoal is used in formulating animal feeds becauseoftheir IJTSRD23724
  • 2. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 418 ability to absorb nitrogen and ammonia and helps in activating the intestinal function of animals (Quiayum et al. 2014). Charcoal is a form of carbon, which is generally produced as a carbonaceous residue of wood and other industrial wastes after heating the organic matter. It contains 70 to 90% pure carbon and several minerals such as calcium, potassium, iron, sodium, copper, zinc, manganese, magnesium, etc. (Brouwer et al.1996).Charcoal has a large surface area giving it countless bonding sites which can be used in adsorbing toxins, gases, and drugs without any specific action (Osol, 1975). It has been used as a feed additive for many terrestrial animals because of its ability to absorb gases especially nitrogen and ammonia, activate the intestinal function, and eliminate the poisons and impurities fromthegastrointestinaltractof land animals (Banner et al. 2000, Bisson et al. 2001, Kutlu et al. 2001, Samanya and Yamauchi 2001, Mekbungwan et al. 2004). It has also been proven to be a good feed additive in the feeds of aquatic animals such as tiger pufferfish (Takifugu rubripes) Japanese flounder (Paralichthys olivaceus), Pangasius hypophthalmus, Clarias gariepinus, Nile tilapia (Oreochromis niloticus), red tilapia hybrid (Oreochromis mossambicus × Oreochromis niloticus) and Gilthead Seabream (Sparus aurata) (Thu et al. 2009, 2010, Quaiyum et al. 2014, Jahan et al. 2014, Lawal et al, 2018, Nopadon et al. 2015, Michael et al. 2015, Michael & Helal 2018). This study aimed to evaluate the effect of pulverized wood charcoal on the on growth, feed utilization, survival and waste excretion of Red tilapia (Oreochromis sp.) MATERIALS AND METHODS Experimental fish and feeds: Red tilapia fingerlings were obtained from Freshwater Aquaculture Centre, Central Luzon State University, Science City of Muñoz, Nueva Ecija, Philippines. Experimental fish with an initialweightof7.32± 1.31 g were acclimatized for one week and were starved for 24 hours before the commencement of the experiment. Ten randomly selected red tilapia were placed in each glass aquarium and fed the with their designated feed thriceaday (8:00, 12:00 and 16:00), with an initial feeding rate of 10% of their body weight. Sampling was done once every two weeks to measure their weight and adjust the feeding rate. This study was designed randomly comprising of 3 treatments with each treatment replicated thrice. Commercially-available diet (35% protein, 4%fat) was used as the control for the experiment. Commercial diet was added with 0% (Control) (T0), 1% (T1) and 2% (T2) PWD and water (200 mL kg-1) (Cho and Lee, 2012). Mixtureswere mixed thoroughly and dried at room temperature before storing at -20 °C. Growth performance and Feed Utilization: To assess the growth performance and feed utilizationof theexperimental fish, the following data weregathered duringtheexperiment which includes specific growth rate (SGR), relative growth rate (RGR), weight gain (WG), survival rate (SR) and feed conversion ratio (FCR). Parameters were calculated as follows: Water Quality Monitoring: Water quality parameterssuch as temperature, dissolved oxygen (DO) and pH were measured using DO meter and pH meter. For the analysis of TAN, water samples were collected fromtheaquariumtanks and were placed in polyethylene (PE) bottles prior to the analysis in the laboratory. Analysis of TAN was done every week before water change following the phenate method. Briefly, 25 to 50 mL of water sample was filtered through Whatman No. 42, and 10 mL of the filtered sample was transferred into a 50 mL beaker in a magnetic stirrer. While stirring, 1 drop of manganese sulfate(MnSO4•H2O)solution, 0.5 mL oxidizing solution and 0.6 mL phenate solution was added. Then, removed from stirrer and allowed 15 minutes for maximumcolor developmentbeforetransferringtransfer to the 1-cm cuvette. Ammonia-free distilled water (10.00 mL) (reagent blank) and 10.00 mL of the 0.300 mg · L-1 solution of total ammonia-nitrogen (standard) were carried through the procedure with each set. With the spectrophotometer at 630 nm, set 0.0 absorbance (100% transmittance) with the reagent blank, the absorbances of the standard and the samples were read. Lastly, the concentrations of the TAN were calculated using the formula: Statistical Analysis: Results for the growth, survival and feed utilization were subjected to one-way analysis of variance (ANOVA) followed by Duncan’s test at a significant level of P<0.05. RESULTS Table1. Growth performance and feed utilization of Red tilapia fed on diets supplemented with different levels of pulverized wood charcoal (PWC) for 35 days. Parameters T0 (0% PWC) T1 (1% PWC) T2 (2% PWC) Initial ABW, g 7.80 ± 0.85a 6.85 ± 2.00a 7.32 ± 1.23a Final ABW, g 14.71 ± 2.72a 15.18 ± 1.45a 13.84 ± 1.85a SGR (%) 0.78 ± 0.14b 1.22 ± 0.10a 0.80 ±0.17b RGR (% · day-1) 87.91 ± 21.79b 168.72 ± 21.99a 91.25 ± 24.52b WG, g 6.91 ± 2.11a 8.33 ± 2.34a 6.52 ± 1.57a FCR 3.72 ± 0.36a 2.45 ± 0.98a 2.98 ± 0.45a SR (%) 36.67 ± 5.77a 46.67 ± 15.27a 53.33 ± 37.85a PWC, pulverized wood charcoal; ABW, average body weight; SGR, specific growth rate; RGR, relative growth rate; WG, weight gain; FCR, feed conversion ratio; SR, survival rate.
  • 3. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 419 Means in column with the same letter are not significantly different at P < 0.05 Figure.1. Total ammonia nitrogen (TAN) co+ncentration of water of aquaria stocked by red tilapia fed on diets supplemented with different levels of pulverized wood charcoal (PWC) for 35 days. Results of growth performance, feed utilization,andsurvival, at the end of the experiment is shown in Table 1. There were no significant differences observed in the initial and final ABW of the experimental fish. However, PWC inclusion of 1% (T1) significantly (P < 0.05) improved the SGR and RGR of the experimental fish whencomparedtothecontrolgroup (T0). WG from T1 was also numerically higher compared to the other treatments (T0 and T2), however, the observed increases were not statistically significant based on comparisons with the other groups. Furthermore, although FCR and SR were better on PWC-supplemented diets, no significant differences were observed upon comparing with the control group. Water quality parameters such astemperature(28.75± 0.85 ºC), pH (8.4 ± 0.22), and DO (2.5 ± 0.83 mg · L-1) did notshow any significant differences among the treatments. Levels of TAN concentration were found to be varying throughoutthe culturing period. During the 1st week until the 3rd week of feeding experiment, a high concentration of TAN was recorded in PWC-supplementeddiets,withreadingsof0.996 to 1.016 mg · L-1, respectively. As the experiment progress, the level of TAN decreases (Fig 1). In the 4th and 5th week of the feeding trial, TAN levels from PWC-supplemented diets were numerically lower, however, the observed decrease was not statistically significant based on comparisons with the control group. DISCUSSION The quality of the fish diet is one of the most important factors to consider when it comes to the optimal growth potential of an aquaculture organism. Over the years, numerous efforts were made to formulate a diet that can provide optimal growth, and made of readily available and inexpensive ingredients to reduce the cost of feed production. However, despite the capabilities of the feeds to sustain good fish growth rates, they have a negative environmental impact due to their high nitrogenous wastes particularly ammonia that constitutes almost 80% of these wastes (Kikuchi et al. 1995). With that, the use of feed additives especially charcoals to reduce the nitrogenous waste coming from the aquaculture industry and promoting better growth to the cultured species is gaining popularity. The inclusion of charcoal in the diets of animals hasreceived attention due to its non-nutritive and nondigestible characteristics giving it the ability to adsorb toxins from the gastrointestinal tract thus preventing their absorption into the body. Several researches have demonstrated that charcoal-supplemented diets can improve fish growth, flesh quality, and intestinal function (Boonanuntanasarn et al. 2014, Quiayum et al. 2014, Jahan et al. 2014, Moe et al. 2010, Thu et al. 2009, 2010, Lawal et al. 2018, Nopadon et al.2015, Michael & Helal 2018). The present study aimed to evaluate the effect of different levels of PWC in the feeds of red tilapia on their growth, feed utilization, survival, and waste excretion. Results showed that 1% inclusion of PWC in the diet significantly improved the SGR and RGR of the fish. The result obtained from this study is in parallel with the report of Nopadon et al. (2015), wherein 1-3% of AC inclusion improved the growth performance of Nile tilapia with optimal inclusion of 2%. Also, Abdel-Tawwab et al. (2017) observed improvement in the growth parameters of Nile tilapia at AC inclusion of 0.2- 2%, with optimal inclusion level of 0.7%. Additionally, the inclusion of commercial wood charcoal(CWC)at3-4%inthe diet of red tilapia hybrid (Oreochromis mossambicus × Oreochromis niloticus) improved fish survival, growth, and feed utilization. In contrast, the present study found no significant improvement in the growth of red tilapia at a 2% level of PWC inclusion. Supplementation of charcoal in the diets have also shown improved growth performance of other aquatic animals. Thu et al. (2009) and Michael & Helal (2018) reported that 4% of charcoal inclusion is the optimal level for tiger puffer fish and gilthead seabream. While 0.5% level of bamboo charcoal (BC) inclusion is reported by Moe et al. (2010) to be the best in juvenile Japanese flounder. In addition, Lawal et al. (2018) found significant improvement in the growth of Clarias gariepinus with 0.25% level of AC inclusion. Furthermore, Quiayum et al. (2014) and Jahan et al. (2014) observed significant effects of BC-supplemented diets in pangasiid catfish growth and ammonia loading, however, they reported a dissimilar level of optimum charcoal inclusion of 2% and 1%, respectively. On the other hand, Boonanuntanasarn et al. (2014), reported that growth parameters of Nile tilapia (O. niloticus) reared on diets containing 10 g · kg-1 body weight AC only showed a numerical improvement with no statistical significance.The same result was observed by Mabe et al. (2017) in juvenile common carp (Cyprinus carpio L.) fed with differentlevels of BC in the diet. These results suggest that the optimal level of charcoal inclusion in the diet of aquaticanimalsdiffer among species, animal’s life stage, and the nature of charcoal. The enhanced growth parameters in the present study may be explained due to the capability of PWC to adsorb and eliminate unwanted gases and toxic substances, which can result to improved digestion, metabolism, and better nutrient utilization and absorption (Kutlu et al. 2001,
  • 4. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID - IJTSRD23724 | Volume – 3 | Issue – 4 | May-Jun 2019 Page: 420 Samanya & Yamauchi 2001, Jahan et al. 2014). Nevertheless, since charcoal is a nonspecific adsorbent, its application in the diets of animals may possibly causetheadsorptionof not only toxins but also nutrients from the gastrointestinaltract. Thus, positive and negative impacts on physiological functions, growth and overall health of an organism can be both expected (Boonanuntanasarn et al.2014,Nopadonetal. 2015). The SR obtained in all treatments is noticeably low. Mortalities of the experimental fish started during the 3rd and 4th week of the experiment. As an observation, some experimental fish are gliding theirbodies alongthesidesand bottom of the aquarium. It is possible that during the experiment, the fish acquired bacterial disease which is one possible reason why the SR in all treatments is low. Aside from that, the level of oxygen throughout the experiment (2.5 ± 0.83 mg · L-1) is lower than the desired dissolved oxygen level of at least 5 mg · L-1. In the present study, no significant improvement was observed in the waste excretion of the red tilapia. In contrast, Thu et al. (2009) and (2010) reported that (BC) supplementation reduced the ammonia-N values with tiger puffer fish and Japanese flounder, respectively.Michaeletal. (2015) also observed a gradual decrease in ammonia levels in the rearing water area as the CWC levels in red tilapia feeds increases. The same observation was noted by Quiayum et al. (2014), where they found out that increasing level of charcoal incorporation in fish feeds results to lower ammonia level in the water. The possible reason with this is that the duration of the experiment (35 days) is not enough to observe the effect of PWC in the waste excretion of the fish. CONCLUSION In conclusion, the result of the present study showed that 1% inclusion of PWC in the diet improved the growth performance of the red tilapia (Oreochromis sp.). Therefore, PWC at 1% can be a useful feed additive in the diet of red tilapia as it is cheap and readily available. ACKNOWLEDGEMENT The author would like to acknowledge the assistance of the College of Fisheries and Freshwater Aquaculture Centre where the study was conducted. Also, the author would like to extend his appreciation to the Department of Science and Technology–Accelerated Science and Technology Human Resource Development Program (DOST-ASTHRDP) for financial assistance during the experiment. REFERENCES [1] Abdel-Tawwab, M., El-Sayed, G.M. & Shady, S.H. 2017. 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