Hematology of Asian redtail catfish (Hemibagrus

nemurus) in different stocking densities using an

aquaponic recirculation system
Niken A. Pamukas, Usman M. Tang, Mulyadi

Department of Aquaculture, Faculty of Fisheries and Marine Science, Riau University,

Pekanbaru, Indonesia. Corresponding author: N. A. Pamukas,
[email protected]

Abstract. Asian redtail catfish (Hemibagrus nemurus) is one of the economically important freshwater
fish commodities in Indonesia and has been popular with domestic consumers in Southeast Asia. The
high consumer demand for this fish incurs the intensification of cultivation, which is carried out with high
stocking densities and high feed consumption. Determining the optimal value for stocking density is a
prerequisite for ensuring growth performance, production, a lower stress level and fish health. Stress
level and fish health can be seen from its hematology. This study aimed to obtain the optimal stocking
density of redtail catfish for culture in a booster system using aquaponic recirculation. The optimal
density was determined based on its hematology (erythrocytes, hematocrit, hemoglobin, leukocrit, and
blood glucose). The research was conducted with a completely randomized design experimental method
with 1 factor, 4 levels of treatment and 3 replications. The treatments consisted of different stocking
densities: 600 fish m-3, 700 fish m-3, 800 fish m-3 and 900 fish m-3. The redtail catfish fingerlings used
had an average weight of 1.60±0.36 g per fish. Maintenance was carried out for 56 days. Analysis of
blood images and blood glucose was carried out on day 1, day 28 and day 56. The results showed that
the different stocking densities had a significant effect (p<0.05) on the hematocrit, hemoglobin, leukocrit
and blood glucose values, but had no effect on erythrocytes and leucocytes values of redtail catfish. The
best stocking density was 700 fish m-3, with the number of erythrocytes of 2.56±0.05 x 106 cells mm-3, a
hematocrit of 31.3±1.5%, hemoglobin value of 10.53±0.47 g dL-1, leucocytes value of 2.23±0.12 x 104
cells mm-3, leukocrit of 1.80±0.10%, blood glucose of 63.33±3.51 mg dL-1, and survival rate of
92.5±1.18%. The range of water quality during the study supported the life of redtail catfish.
Temperatures ranged from 26 to 28.20C, pH was between 6-7.4, dissolved oxygen values were between
7-7.7 mg L-1 and ammonia values were between 0.0003-0.0257 mg L-1.
Key Words: filter, fish health, survival rate, water quality.

Introduction. Redtail catfish (Hemibagrus nemurus) is one of the economically

important freshwater fish commodities in Indonesia and has been popular with domestic
consumers in Southeast Asia. Redtail catfish are native fish in inland waters that can live
in lakes, rivers and swamps. In Indonesia, the fish inhabits the Kampar River in Riau
Province (Husnah et al 2003), Barito River in Kalimantan (Samuel & Said 1995),
Batanghari River in Jambi (Nurdawati et al 2006), Musi River in South Sumatra
(Muflikhah et al 2006), Lake Singkarak in West Sumatra (Uslichah & Syandri 2003),
among others.
The redtail catfish has a good meat quality, so the price is high compared to other
freshwater fish (Boyoun et al 2010). This fish is preferred by the community because it is
thick fleshed, slightly prickly and it has a delicious taste. The demand from Malaysia and
Singapore is also high (Aryani et al 2002). The price of this fish can reach 5.15-5.49 USD
kg-1 in fresh form, while for processed smoked fish the price is 17.17-27.47 USD kg-1.
However, commercialization and intensive farming of this species is still limited, due to
the inability of the fish to breed naturally in captivity and slow growth; the provision of
juvenile stages for rearing is still largely relying on capture from the wild (Roza et al
2014).
The high consumer demand for redtail catfish needs an intensification of
cultivation, which is carried out with high stocking densities and large amounts of feed.

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1534
The application of booster system of fish culture with an aquaponic recirculation system
can increase the production of redtail catfish. A booster system of fish culture is a super
intensive culture system that applies a high stocking density technology using the
application of a booster supplement product that contains nutrients, minerals and
probiotics to improve water quality, feed and fish immunity. In order for the cultivation
system to work optimally, it is necessary to have an appropriate stocking density of
redtail catfish. Determining the optimal value for stocking density is a prerequisite for
ensuring growth performance, production, low stress level and fish health. Stress level
and fish health can be seen from its hematology. Hastuti & Subandiyono (2015) reported
that African catfish (Clarias gariepinus) reared at high a stocking density (1.6 kg m-3)
showed a decrease in the number of erythrocytes, leukocrit, hemoglobin and
haematocrit, which were below normal standards. However, there was an increase in the
platelet count, which was above the normal range, indicating that the fish was under
stress.
The information on high densities on the health of redtail catfish, especially on its
blood profile is scarce. For this reason, this study was conducted with the aim of
obtaining the optimal stocking density in the cultivation of the booster technique with the
aquaponic recirculation system, verified with the blood profile.

Material and Method

Time and Location of the study. This research was conducted from March to
September 2020 at the Hatchery Technical Implementation Unit and Experimental Pool of
the Faculty of Fisheries and Marine Sciences, Riau University. The determination and
interpretation of fish blood profiles was carried out at the Laboratory of Parasites and Fish
Diseases, Faculty of Fisheries and Marine Sciences, Riau University, Pekanbaru.

Materials and instruments used. The materials used in this study were: 1000 redtail
catfish juveniles with a weight ranging from 1.6±0.36 g per fish, which came from
Kampar Regency. The test feed used was commercial pellet with 39% crude protein, 5%
crude lipids, 16% ash, 12% moisture and 22% NFE, with the production code FF-999
from Charoen Pokphand Inc. The bok choy (Brassica chinensis L.) seeds used for the
aquaponic system came from vegetable farmers on Kertama Street, Pekanbaru. Materials
used for blood profile determination and interpretation were Hayem diluting fluid (1:200),
Turk blood diluting liquid (1:20), 10% EDTA, 0.1 N HCl and distilled water. The tools used
in this study were: a round tube test container with a diameter of 60 cm, a height of 45
cm, a volume of 100 L, an analytical scale with an accuracy level of 0.1 mg, a binocular
microscope, a microhematocrit centrifuge model SH 120-1, a syringe, a hematocrit
capillary, hematocrit reader, shelfs, Eppendorf tubes, gluco Dr, slides, cover glass,
salinometer, crystoseal, hemocytometer, pH meter, DO meter, refractometer and
spectrophotometer.

Experimental design. The research method used in this study was an experimental
method using a completely randomized design (CRD) with 1 factor, 5 levels of treatment
and 3 replications (Steel et al 1996). The variable in this study was the different stocking
density. The treatment in this study referred to the best treatment obtained by
Matondang et al (2019) in a booster system, namely the best stocking density of redtail
catfish of 700 fish m-3 (1.12 kg m-3). Thus, the treatments in this study consisted of:
P1=600 fish m-3 (0.978 kg m-3), P2=700 fish m-3 (1.169 kg m-3), P3=800 fish m-3 (1.28
kg m-3), and P4=900 fish m-3 (1.377 kg m-3).

Preparation of test containers. The containers used in this study were 15 units of
round plastic tubes with a diameter of 60 cm and a height of 45 cm, with a maximum
water capacity of 100 L. Before use, the containers were washed using booster blue
copper (each liter containing: 200 mg copper sulfate, 25 mg cetyl pyrinidium chloride,
and 20 mg cetyl trimethyl ammonium bromide) at a dose of 1 ppm, and then dried. Next,
the containers were filled with 80 L of water, and 1 ppm of booster blue copper was

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1535
added in order to conduct water sterilization (Wulandari 2014). The maintenance bath
was equipped with a 32 watt pump to drain water from the maintenance container to the
filter housing.

Preparation of bok choy plant seeds and filter containers. Bok choy seedlings
preparation consisted in planting bok choy seedlings into rockwool measuring 3x3 cm 2,
which had been perforated using a stick; each hole was filled one bok choy seedling
(Zalukhu et al 2016). Seedlings used were not defective and were perfectly open, ranging
between 4 and 4.5 cm in height, with 3 leaves each. Furthermore, each seedling was
transferred to the netpot, which contained zeolite at the bottom, with rockwool on top.
The amount of zeolite in each netpot was the same, 30 g. The zeolite was expected to
purify water.
The assembly of the aquaponics system was conducted by making a hole in the
maintenance container according to the size of the pipe, then connecting it to the gutter.
The gutter used had 15 units with the size of 100x13.5x10.5 cm3. Each gutter unit was
made of 10 holes with a distance of 5 cm as a place for placing the plant netpot. Then
the connection was made from each gutter using a paralon pipe to permit water flow.
The fish rearing container was equipped with a water pump with a power of 32
watts to drain the water into the fish tank. The filter tube used was a water gutter with a
volume of 7 L. Furthermore, the water from the filter tube would flow through a PVC pipe
with a diameter of 2.5 cm into the reservoir (plastic container with a volume of 10 L).
The water collected in the reservoir was pumped into the maintenance tank with a
pumping flow of 2 L per minute. Preliminary experiments were carried out to verify that
the recirculation system for fish farming was working properly. In the preliminary
experiment, the parameters observed were the rate of water change in the tank and in
the filter (Putra & Pamukas 2011). After all the containers had been assembled, they
were labelled according to treatment (Figure 1).

Figure 1. Container for fish rearing with the booster technique using the Brassica
chinensis L. aquaponic recirculation system.

Preparation of media for fish farming. The growth of plankton in the redtail catfish
rearing media was stimulated by adding a fermentation mixture made from 0.5 kg of
bran, 200 cc of plankton booster, 10 g of aquaenzyme booster and 20 cc of liquid amino
booster, incubated for 36 hours. The fermentation product was then dissolved in 10 L of
water, and filtered, then 1.35 mL L-1 of suspension was put into the maintenance medium
in the morning. This was repeated every 9 days (Pamukas et al 2020). Powerful Booster
(Manstap with P2O5, KNO3, SiO2, and trace elements Ca, Mg, Co, Cu, Fe, Mn, Se, Zn) at a
dose of 30 ppm was added in the afternoon after the distribution of the fermented
booster suspension in the morning. The addition of water was performed after

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1536
administering the booster staple until the volume of water in the container reached 80 L
(35 cm high).

Preparation of test fish. The criteria for the fingerlings selected as test animals were
1.6±0.36 g per fish in size, active, not defective, and relatively uniform in size. The
fingerlings were stocked in densities according to treatments, converted to the volume of
water in the containers: P1=600 fish m-3 (0.978 kg m-3), P2=700 fish m-3 (1.169 kg m-3),
P3=800 fish m-3 (1.280 kg m-3), and P4=900 fish m-3 (1.377 kg m-3).
The fingerlings were soaked in a booster fish immunovit solution at a dose of 0.6
mL L-1 for 15 minutes. A stress off booster was applied with a dose of 2 mL for 5 L of
water, then acclimatization was carried out by inserting a plastic bag containing the fish
into the rearing container for 1-2 hours to adjust the water temperature.

Feeding. The frequency of feeding in this study was carried out 3 times a day, at 08.00,
13.00 and 16.00, with 3% of fish weight per day (Affandi et al 2009).
The supplements used from the first day to the 30th day of maintenance were
liquid amino booster 5 mL kg-1 of feed, booster grotop 2 g kg-1 of feed, and booster
premix aquavita 2 g kg-1 of feed. Meanwhile, on the 31st day until the end of
maintenance, supplements used were liquid amino booster 5 mL kg-1 of feed, booster
grotop 2 g kg-1 of feed, booster vitaliquid 2 g kg-1 of feed (Sudarmaji & Boster 2013).
Addition of supplements to feed was done by mixing all booster supplements
according to the specified dose, then dissolved with water (100 mL kg-1) and sprayed on
the feed evenly and dried. In addition, the feed was stored in a clean and dry container.
Each feed was weighed according to the body weight of the fish. The feed was given in an
evenly distributed manner, so that each fish had equal access to food.

Maintenance. Redtail catfish were kept for 56 days. The length and weight of fish were
determined every 14 days using an analytical balance with an accuracy of 0.01 and a
gauge. Weight measurements were carried out to adjust the dose of feed administered.
The survival rate was known by counting the number of fish dying each day during the
rearing period. For 3 consecutive days after the fish were stocked and once a week
during the maintenance period, multi-cell boosters were stocked at a dose of 20 ppm
during the day.

Redtail catfish blood collection. Asian Redtail catfish blood sampling was conducted at
the beginning of the study and after 56 days of maintenance. 3 fish samples were
colelcted from each treatment and each replication. The observed hematology included
the number of erythrocytes, hematocrit, hemoglobin, leukocrit, and blood glucose. The
fish blood was collected by anesthetizing the test fish using clove oil (0.1 mL L-1). The
syringe and microtube used to draw blood were rinsed with 10% EDTA. Blood was taken
using a 1 mL syringe from the caudal vein with a tilt angle of 45 0, the blood was inserted
into a microtube (1 mL of blood could be collected), then put into a basin filled with ice
cubes, so that the blood did not clot and was immediately observed in the laboratory.

Measured response. The parameters measured in this study were: total erythrocytes
according to Klontz (1994), hematocrit and leukocrit levels according to Anderson &
Siwicki (1995), hemoglobin level using the Sali method with a salinometer according to
Wedemeyer & Yasutke (1977), total leukocrit according to Blaxhall & Daiesley (1973),
blood glucose level according to Eames et al (2010), fish survival according to Effendie
(1986), and water quality (temperature, pH, dissolved oxygen (DO), ammonia (NH3),
nitrite (NO2), and nitrate (NO3) were measured every week during the study according to
Indonesian National Standard (SNI) in the Public Works Service (1990).

Data analysis. Data on total erythrocytes, level of hematocrit, leucocrit, hemoglobin,

total leukocrit, blood glucose and fish survival during the study were analysed according
to a completely randomized design model (Steel et al 1996). To determine the effect of
the treatments on each measured variable, the analysis of diversity was carried out using

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1537
the F statistical test. The analysis process used SPSS version 13.0 software. If p<0.05,
then the Newman-Keuls test was performed to see the differences among treatments.
Water quality was analyzed descriptively.

Results and Discussion

Erythrocytes. The mean total erythrocytes at the beginning of the study (day 1) was
1.92 x 106 cells mm-3. On day 28, the mean total erythrocytes increased in all
treatments until day 56 (Figure 2). This was because the fish had adapted to the
medium, so that stress level decreased and appetite increased, which was followed by an
increase in erythrocytes. The highest increase in total erythrocytes was found at a
stocking density of 700 fish m-3, 2.09x106 cells mm-3 (day 28) and 2.56x106 cells mm-3
(day 56). This was because the stocking density was the optimal stocking density from
the tested densities for redtail catfish, so that fish were in a comfortable condition and
had an impact on improving health and ultimately increasing the number of erythrocytes.
Addini et al (2020) found that the number of erythrocytes in the stocking density
of fish of 250 fish m-3 was greater than the number of erythrocytes at the stocking
density of 200 fish m-3 and 300 fish m-3. According to Harianto et al (2014), an increase
in the erythrocyte level is caused by the physiological adaptation of the fish to the
homeostatic conditions; the increased erythrocyte value indicates that the fish are in a
healthy status. Erythrocyte levels were higher at the stocking density of 700 fish m-3
compared to 600 fish m-3. It is assumed that the redtail catfish likes to school, so that
the stocking density is suitable for the fish.
The total erythrocyte values at the stocking densities of 800 fish m-3 and 900 fish
m were lower compared to 700 fish m-3. This is because at a high stocking density, fecal
-3

waste and metabolism will increase, so that the ammonia content will be higher than at
lower stocking densities. This is in line with the opinion of Ni et al (2014), who stated
that the number of erythrocytes is influenced by the density of distribution. The high
stocking density caused the ammonia content in the culture medium to be higher.
Furthermore, the performance of cells in the blood of African catfish was influenced by
the quality of the water media, especially ammonia. According to Li et al (2013), catfish
(Pelteobagrus vachelli) reared in high ammonia (5.7 mg L-1 TAN) contained a lower
number of erythrocyte cells, hemoglobin and hematocrit compared to their values in fish
raised in ambient ammonia (0.01 mg L-1 TAN). The erythrocyte value of P. vachelli
cultured in high ammonia (5.7 mg L-1 TAN) was 2.28±0.07 (x106 cells µl-1) and the
catfish erythrocyte value in low ammonia was 2.7±0.08 (x106 cells µl-1).

Figure 2. Average total erythrocytes at different stocking densities of fish.

The total erythrocyte level in all treatments during the study was in the normal range for
the redtail catfish (1.92 - 2.56x106 cells mm-3). This is in accordance with the opinion of

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1538
Fitria et al (2019), who reported that the number of normal erythrocytes in redtail catfish
living in their natural habitat in Ombilin and Sawahlunto were 2.268x106 cells mm-3 and
2.267x106 cells mm-3, respectively. Furthermore, Irianto (2005) states that the number
of normal erythrocytes in teleostei fish ranges from 1.05 to 3x106 cells mm-3.
The results of analysis of variance (ANOVA) showed that the stocking density had
a significant effect (p<0.05) on the number of erythrocytes on day 56, while on days 1
and 28 there were no significant differences. The Newman-Keuls method (SNK) showed
that the number of erythrocytes at a stocking density of 700 fish m-3 was different from
the number of erythrocytes in other treatments on the 56th day. According to Emu
(2010), the factors influencing the number of erythrocytes are species, sex, age, dietary
nutrition and size. In addition, according to Yanto et al (2015), environmental conditions
such as lack of oxygen and high temperatures can cause a decrease in the number of
erythrocytes. In the study, although the test fish used were of the same age and size,
and during culturing period the environmental conditions were relatively homogeneous,
in the first 2 weeks the fish still adapted to the environment, feed and stocking density,
so the number of erythrocytes did not increase significantly.

Hematocrit. The hematocrit level of redtail catfish increased from the beginning (day 1)
to the end of the study (day 56). The highest increase in hematocrit occurred at a
stocking density of 700 fish m-3, while the lowest was found at stocking density of 900
fish m-3 (Figure 3). The hematocrit level is the ratio between red blood cells and blood
plasma, so that the number is related to the number of erythrocytes (in our case
depending on stocking density). Fitria et al (2019) state that a high number of
erythrocytes will be followed by an increased percentage of hematocrit. According to
Nuryati et al (2006), the hematocrit level is related to the number of red blood cells.
Furthermore, Fadil et al (2011) state that an increase in the level hematocrit in fish blood
indicates a relationship with the large number of red blood cells formed by the
haematopoiesis tissue, the number of red blood cells being directly proportional to the
hematocrit value. Setiawati et al (2017) explain that a low hematocrit count indicates a
lack of food or decreased appetite due to stress.

Figure 3. Average level of hematocrit at different stocking density of redtail catfish

(Hemibagrus nemurus).

Based on the ANOVA test, the stocking density had an effect on the hematocrit level
(p<0.05). The SNK test showed that the hematocrit value at the stocking density of 700
individuals m-3 was significantly different from the hematocrit value at the stocking
densities of 600, 800 and 900 individuals m-3. The hematocrit level of redtail catfish at

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1539
the beginning of the study was low, 17.2%, indicating that the fish were in the process of
adapting to their environment. On day 28 and 56, the hematocrit levels increased to 19-
31.3%. According to Bastiawan et al (2001), the hematocrit values in normal fish range
from 30.8-45.5%. The hematocrit values at the stocking densities of 600, 800 and 900
fish m-3 were classified as low, presumably because the stocking density was not optimal,
so that the fish were stressed and their appetite decreased. At the stocking density of
700 fish m-3, the hematocrit value was classified as normal, indicating that the fish were
healthy and the stocking density was optimal.

Hemoglobin. Hemoglobin levels in all treatments increased from baseline to the end of
the study (day 56). The mean range of hemoglobin levels in the fish at the start of the
study was low (5.4 g dL-1), but on days 28 and 56 it increased within normal ranges (7-
10.53 g dL-1). According to Hardi et al (2011), normal hemoglobin levels in fish range
from 6 to 13 g dL-1. The increase in hemoglobin levels is due to the fish adapting well to
the media. The use of the booster technique with aquaponic recirculation causes the
water quality to be maintained in a good range for the life of the redtail catfish (Table 1),
so that the fish is healthy, resulting in an increase in hemoglobin. According to Kurniawan
(2019), hemoglobin levels in the blood are influenced by several factors, including water
quality and oxygen content. DO in this study was classified as high, namely 7–7.7 mg L-1.

Figure 4. Average levels of hemoglobin at diffrent stocking densities of redtail catfish

(Hemibagrus nemurus).

Figure 4 shows that the highest hemoglobin levels were found at the stocking density of
700 individuals m-3 and the lowest at 600 individuals m-3. The ANOVA test results showed
that the stocking density had an effect on the hemoglobin level of the fish (p<0.05). The
SNK test showed that the hemoglobin level at the stocking density of 700 individuals m-3
was significantly different from the other treatments. This indicates that the stocking
density of 700 fish m-3 is the optimal stocking density from the tested densities for redtail
catfish. Caruso et al (2005) and Harianto et al (2014) stated that hemoglobin transports
oxygen throughout the body. Hemoglobin releases oxygen to cells and binds to carbon
dioxide. The amount of oxygen received by the tissue depends on the level and function
of available hemoglobin. If the oxygen level is low, the ability of hemoglobin in blood
transport will decrease. DO content during the study ranged from 7–7.7 mg L-1, this
value strongly supporting the growth of redtail catfish, because it is in the normal range
of more than 3 ppm (Boyd 2015). The value of hemoglobin and oxygen levels produced
in this study indicate that the red tail catfish are in a normal health status.

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1540
Leucocytes. The total leukocyte value during the study ranged from 2.15 to 3.32x104
cells mm-3 (Figure 5), which was in the normal range, which is 2-15x104 cells mm-3
(Royan et al 2014). The total leukocyte value in this study was similar to that obtained by
Fitria et al (2019) from redtail catfish in their natural habitats, ranging from 2.7 to
3.5x104 cells mm-3. This indicates the fish was in good health. During the study, the total
leucocytes value of redtail catfish decreased from day 1 to 56. The decrease in total
leukocytes was thought to be influenced by the production of anti-stress substances by
probiotic organisms in the fish body, as reported by Mohapatra el al (2014). The decrease
in total leukocytes during the study indicated a decrease in stress level due to fish being
able to adapt to its environment. According to Fitria et al (2019), the total leukocyte
value can be influenced by various factors, including water quality and pathogens in the
maintenance media. The water quality during the study was in a good range for the
health of redtail catfish due to the use of the aquaponic recirculation system booster
technique in this study.

Figure 5. Average leukocyte levels at different stocking densities of redtail catfish

(Hemibagrus nemurus).

The results of the analysis of variance (ANOVA) on total leukocyte count during the study
showed an effect of stocking density on total leukocytes (p<0.05). Based on the SNK
test, it was found that the total leukocyte at the stocking densities of 600 and 700 fish m-
3
was relatively the same, but different compared to the stocking densities of 800 and
900 fish m-3. The lowest total leukocyte value was found in the treatment with 600 fish
m-3, and the highest in the treatment with 900 fish m-3. This indicates that at low
stocking densities, the total leukocyte count tended to be lower. This is in line with the
results of Caruso et al (2005) and Haenen et al (2010), who determined that the value of
leukocytes tends to decrease along with the decreasing stocking density. This indicates
that the fish did not experience phagocytosis or environmental stressors causing
increased levels of leukocytes. The increase in total leukocyte count can be caused by
diseases, infections, parasites, stress due to handling and environmental influence. A low
leukocyte value indicates that the fish was in good health or had a good immune
response.

Leukocrit. The leukocrit level decreased from the beginning to the end of the study.
Leukocrit levels in all treatments were high (3.24%) at the beginning of the study, but
decreased later, so that they reached the normal range (1.7 - 2.83%). The leukocrit level
tended to decrease with decreasing stocking density of redtail catfish (Figure 6). Aisiah et
al (2011) state that a normal leukocrit level ranges for catfish (Pangasius
hypophthalmus) ranges from 0.72 to 2.89%. According to Hastuti (2007), a low leukocrit
level can be caused by chronic infections, low nutritional quality, lack of vitamins and the
presence of contaminants. Meanwhile, an increase in leukocrit can indicate an infection at

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1541
an early stage or that the fish is under stress. The leukocrit value of fish is also
dependent on the condition of the fish at the time of sampling, the period of time
between sampling and blood measurement, and the measurement procedure used. The
decrease in leukocrit level indicates that the fish were not under stress and were not
attacked by pathogens. The leukocrit levels were in the normal range.
Based on the results of the analysis of variance (ANOVA), it was found that the
different stocking densities had an effect on the leukocrit level of the fish (p<0.05). The
leukocrit level in the stocking density of 700 fish m-3 was different from those in other
stocking densities. This shows that the stocking density of 700 fish m-3 was the optimal
stocking density for redtail catfish.

Figure 6. Average leukocrit levels at different stocking densities of redtail catfish

(Hemibagrus nemurus).

Blood glucose. The range of fish blood glucose decreased from day 1 to day 56, except
in the stocking density of 900 fish m-3, where it increased to day 28 and decreased to day
56 (Figure 7).

Figure 7. Average blood glucose levels at different stocking densities of redtail catfish
(Hemibagrus nemurus).

Figure 7 shows that the stocking density had an effect on the blood glucose level
(p<0.05). Based on the SNK test, it was found that the blood glucose of the fish in the
stocking densities of 600, 700 and 800 fish m-3 were relatively the same on the 28th day,
but different from the stocking density of 900 fish m-3. The blood glucose on day 56
differed between treatments, except in the stocking densities of 800 and 900 fish m-3,

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1542
where they were relatively the same. The blood glucose range in all stocking densities
was within the normal range (56-84.33 mg dL-1). Blood glucose levels decreased with the
lower stocking density. This is in accordance with the research of Ajani et al (2015), who
stated that the glucose level will increase when the fish experiences stress, directly
proportional to the increase in stocking density used. According to Nasichah et al (2016),
normal fish glucose level is between 40-90 mg dL-1, whereas Hartanti et al (2013) give a
different normal blood glucose in fish, between 41-150 mg dL-1. Van de Nieuwegiessen et
al (2009) reported that Clarias gariepinus responds to stress by increasing its blood
glucose level. Putra et al (2020) state that stress hormones adrenaline and non-
adrenaline are associated with cortisol secretion and glucose fluctuation to control energy
use at high stocking densities.

Water quality. The temperature range, pH, DO and ammonia in all treatments during
the study were relatively similar, except the highest ammonia concentration, which was
found at the stocking density of 900 fish m-3 (Table 1). The water quality in all
treatments was classified as good for the life of the redtail catfish. According to Tang
(2003), the optimal temperature for the growth of redtail catfish is in the range of 27-
330C. The optimal pH should be between 5.9-6.4, DO should be between 5.6-7.4 mg L-1
(Roza et al 2014), and ammonia should be below 0.02 mg L-1 (Saputra et al 2019). The
water quality was maintained relatively constant during the study due to the application
of the booster technique with aquaponic recirculation.

Table 1
Water quality for redtail catfish (Hemibagrus nemurus) at different stocking densities

Treatment Temperature (oC) pH DO (mg L-1) Amonia (mg L-1)

600 fish m-3 26.1–28.2 6–7.4 7–7.7 0.0003–0.0058
(0.978 kg m-3)
700 fish m-3 26.1–28.2 6–7,4 7–7.7 0.0003–0.0052
(1.169 kg m-3)
800 fish m-3 26.1–28.1 6–7.4 7–7.7 0.0003–0.0076
(1.280 kg m-3)
900 fish m-3 26–28.2 6–7.3 7–7.7 0.0003–0.0257
(1.377 kg m-3)

The survival rate. The average survival rate of redtail catfish in all treatments was high
(79.9-92.52%). The highest survival rate was found at stocking density of 700 fish m-3
and the lowest was at the stocking density of 900 fish m-3 (Figure 8). The high survival
rate at a stocking density of 700 fish m-3 occurred because the hematological conditions
of the redtail catfish were at best values in this treatment compared to other treatments.

Figure 8. Fish survival rate at different stocking densities.

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1543
ANOVA results showed that the stocking density had an effect on the survival of the
redtail catfish (p<0.05). Based on the SNK test, the survival rate was different among
different stocking densities. This is because the hematological conditions of the redtail
catfish were also different at different stocking densities. The survival rate in this
treatment (1.169 kg m-3) was higher than that obtained by Matondang (2019), which
was 77.08-91.66%. This is due to the application of the aquaponic recirculation system
booster technique, which caused water quality to remain in a good range for the growth
and survival of the redtail catfish.

Conclusions. Redtail catfish stocking density affected total erythrocytes, hematocrit

level, hemoglobin level, total leukocytes, leukocrit level, blood glucose and survival rate.
The best stocking density was at 700 fish m-3 (1.169 kg m-3 – 5,096 kg m-3), with the
following values: number of erythrocytes: (2.56±0.05)x106 cells mm-3; hematocrit:
31.3±1.5%; hemoglobin: 10.53±0.47 g dL-1; leukocytes: (2.23±0.12)x104 cells mm-3;
leukocrit: 1.8±0.1%; blood glucose: 63.33±3.51 mg dL-1; survival rate: 92.5±1.18%.
The water quality during the study supported the life of redtail catfish.

Acknowledgements. The author would like to thank the Institute for Research and
Community Service and the Ministry of Education and Culture for funding this research.

Conflict of Interest. The authors declare that there is no conflict of interest.

References

Addini N., Tang U. M., Syawal H., 2020 [Physiological growth of lais fish (Ompok
hypophthalmus) in aquaculture recirculation system (ARS)]. Jurnal Berkala
Perikanan Terubuk 48(2):1-15. [In Indonesian].
Affandi R., Syafei D. S., Rahardjo M. F., 2009 [Fish physiology: Digestion and food
absorption]. IPB Press, Bogor, Indonesia, 240 p. [In Indonesian].
Aisiah S., Muhammad, Anita, 2011 [Use of betel leaf extract (Piper betle Linn) to inhibit
Aeromonas hydrophila and its toxicity in catfish (Pangasius hypophthalmus)]. Jurnal
Fish Science 1(2):190-201. [In Indonesian].
Ajani E. K., Setufe S. B., Oyebola O. O., 2015 Effect of stocking density on
haematological functions of juvenile African catfish (Clarias gariepinus) fed varying
crude protein levels. African Journal of Food Science 9(2):65-69.
Anderson D. P., Siwicki A. K., 1995 Basic hematology and serology for fish health
programs. Shariff M., Arthur J. R., Subasinghe J. P. (eds), Fish Health Section,
Asian Fisheries Society, Manila, Phillipines, pp. 185-202.
Aryani N., Syawal H., Bukhari D., 2002 [Testing the use of LHRH hormone for the
maturation of the gonads of catfish (Mystus nemurus, C.V)]. Torani 12(3):163-168.
[In Indonesian].
Bastiawan D. A., Wahid, Alifudin M., Agustiawan I., 2001 [Blood profile of African catfish
(Clarias spp.) infected with Aphanomyces sp. at different pH]. Jurnal Penelitian
Indonesia 7(3):44-47. [In Indonesian].
Blaxhall P. C., Daisley K. W., 1973 Routine haematological methods for use with fish
blood. Journal of Fish Biology 5(6):771-781.
Boyd C., 2015 Water quality. In: Aquaculture: Farming aquatic animals and plants. Lucas
J. S., Southgate P. C. (eds), Blackwell Publising, Chichester, pp. 52-83.
Boyoun H. S., Catur S., Yudi Y., 2010 [Easy ways to cultivate redtail catfish and Hoven’s
carp fish business opportunities]. IPB Press, Bogor, Indonesia, 161 p. [In
Indonesian].
Caruso G., Genovese L., Maricchiolo G., Modica A., 2005 Haematological, biochemical and
immunological parameters as stress indicators in Dicentrarchus labrax and Sparus
aurata farmed in off-shore cages. Aquaculture International 13:67-73.
Eames S. C., Philipson L. H., Prince V. E., Kinkel M. D., 2010 Blood sugar measurement
in zebrafish reveals dynamics of glucose homeostasis. Zebrafish 7(2):205-213.

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1544
Effendie M. I., 1986 [Fisheries biology]. Pustaka Nusantara Foundation, Yogyakarta,
Indonesia, 163 p. [In Indonesian].
Emu S., 2010 [The use of salt in a closed system of high density catfish (Pangasius sp) in
a medium containing zeolite and activated charcoal]. Thesis, Graduate School, IPB
Bogor, Bogor, Indonesia. [In Indonesian].
Fadil M. S., Syaifullah, Zakaria I. J., 2011 [Study of several aspects of water chemistry
physics and fish physiological aspects found in a rubber factory’s exhaust stream in
the Batang Arau River]. Thesis, Graduate Program Andalas University, Padang,
Indonesia. [In Indonesian].
Fitria N., Tjong D. H., Zakaria I. J., 2019 [Blood physiology of redtail catfish (Hemibagrus
nemurus Blkr)]. Jurnal Metamorfosa 6(1):33-38. [In Indonesian].
Haenen O. L. M., Lehmann J., Engelsma M. Y., Stürenberg F. J., Roozenburg I.,
Kerkhoff S., Breteler J. K., 2010 The health status of European silver eels Anguilla
anguilla in the Dutch River Rhine Watershed and Lake IJsselmeer. Aquaculture
309(1-4):15-24.
Hardi E. H., Sukenda, Harris E., Lusiastuti A. M., 2011 [Characteristics and pathogenicity
of β-hemolytic and non-hemolytic Streptococcus agalactiae in tilapia]. Jurnal
Veteriner 12(2):152-164. [In Indonesian].
Harianto E., Budiardi T., Sudrajat A. O., 2014 [The growth performance of Anguilla
bicolor at initial weight of 7 g at different densities]. Journal of Indonesian
Aquaculture 13(2):120-131. [In Indonesian].
Hartanti, Siwi, Hastuti S., Sarjito, 2013 [Performance profile of blood for African catfish
(Clarias gariepinus) after maintenance with the addition of vitamin C in the feed].
Journal of Aquaculture Management and Technology 2(1):113-125. [In
Indonesian].
Hastuti S. D., 2007 [Evaluation of non-specific defence of tilapia (Oreochromis sp)
injected with LPS (lipopolysaccharides) of Aeromonas hydrophilla]. Jurnal Protein
14(1):79-84. [In Indonesian].
Hastuti S., Subandiyono, 2015 [The health condition of African catfish (Clarias
gariepinus, Burch) maintained using biofloc technology]. Jurnal Saintek Perikanan
10(2):74-79. [In Indonesian].
Husnah S., Aida N., Gautama S., 2003 [Research on the number, type of distribution and
role of cultivated fish regardless of the catch of fish in public waters. Final report on
fisheries technology mastery project]. Balai Riset Perikanan Perairan Umum, 19 p.
[In Indonesian].
Irianto A., 2005 [The pathology of teleostei fish]. Gadjah Mada University Press,
Yogyakarta, Indonesia, pp. 16-54. [In Indonesian].
Klontz G. W., 1994 Fish hematology. In: Techniques in fish immunology. Stolen J. S.,
Flecher T. C., Rowely A. F., Zelikoff T. C., Kaattari S. L., Smith S. A. (eds), SOS
Publications, Fair Haven, USA, pp. 121-132.
Kurniawan A. J., 2019 [Hematological profile of African catfish (Clarias gariepinus) raised
using biofloc system]. Jurnal Perikanan Universitas Riau, pp. 1-9. [In Indonesian].
Li M., Li E., Qin J. G., Chen L., Yu N., Du Z., 2013 Growth performance, antioxidant
status and immune response in darkbarbel catfish Pelteobagrus vachelli fed
different PUFA/vitamin E dietary levels and exposed to high or low ammonia.
Aquaculture 406-407:18-27.
Matondang P. A. S., Tang U. M., Putra I., 2019 [Maintenance of redtail catfish
(Hemibagrus nemurus) in different stocking densities in the boster cultivation
system]. Jurnal Online Mahasiswa, Universitas Riau, pp. 1-14. [In Indonesian].
Mohapatra S., Chakraborty T., Prusty A. K., PaniPrasad K., Mohanta K. N., 2014 Benefical
effect of dietary probiotics mixture on hemato-immunology and cell apoptosis of
Labeo rohita fingerlings reared at higher water temperatures. PLoS ONE
9(6):e100929, 9 p.
Muflikhah N., Nurdawati S., Aida S. N., 2006 [Prospect of river catfish (Mystus nemurus
C.V.) germplasm development]. Bawal 1(1):11-17. [In Indonesian].
Nasichah, Zahrotun, Widjanarko P., Kurniawan A., Arfiati D., 2016 [Analysis of blood
glucose level of Java barb fish (Barbonymus gonionotus) from the downstream

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1545
 Songo weir of the Brantas River]. Brawijaya University, Malang, 333 p. [In
Indonesian].
Ni M., Wen H., Li J., Chi M., Bu Y., Ren Y., Zhang M., Song Z., Ding H., 2014 The
physological performance and immune responses of juvenile Amur sturgeon
(Acipenser schrenckii) to stocking density and hypoxia stress. Journal of Fish &
Shellfish Imunolgy 36(2):325-335.
Nurdawati S. N., Muflikhah M. S., Sunarno J., 2006 [Fishery resources of the Batanghari
River, Jambi]. Jurnal Bawal 1(1):1-9. [In Indonesian].
Nuryati S., Kuswardani Y., Hadiroseyani Y., 2006 [Effect of bee resin on blood profiles of
infected Carassius auratus by Aeromonas hydrophila]. Jurnal Akuakultur Indonesia
5(2):191-199. [In Indonesian].
Pamukas N. A., Syafriadiman, Lukistyowati I., Efriyeldi, Feliatra, Mulyadi, Syakti A. D.,
Fauzi M., Windarti, 2020 Types and abundance of plankton in the hybrid tilapia
brackish water culture media enriched with mixed booster (plankton, aqua enzyme
and amino liquid). AACL Bioflux 13(2):1198-1210.
Putra I., Effendi I., Lukistyowati I., Tang U. M., Fauzi M., Suharman I., Muchlisin Z. A.,
2020 Effect of different biofloc starters on ammonia, nitrate, and nitrite
concentrations in the cultured tilapia Oreochromis niloticus system. F1000Research
9:293, 13 p.
Putra I., Pamukas N. A., 2011 [Maintenance of river catfish (Ompok sp.) in a recirculating
aquaponic system]. Jurnal Perikanan dan Kelautan 16(1):125-131. [In Indonesian].
Royan F., Rejeki S., Haditomo A. H. C., 2014 [The effects of different salinities on blood
profile parameters of tilapia (Oreochromis niloticus)]. Jurnal Teknologi dan
Manajemen Akuakultur 3(2):109-117. [In Indonesian].
Roza M., Manurung R., Budhi A., Sinwanus, Heltonika B., 2014 [Study of the
maintenance of redtail catfish (Hemibagrus nemurus) with different stocking
densities in floating net cages in Sungai Paku Reservoir, Kampar Regency, Riau
Province]. Acta Aquatica 1(1):12-62. [In Indonesian].
Samuel, Said A., 1995 [Relationship of weight and length and condition factor of the river
catfish (Mystus numerus CV) in the Batanghari watershed]. Seminar on the
Compilation of Fisheries Research Results in Public Waters, Department of
Agriculture, Jakarta, Indonesia, pp. 127-130. [In Indonesian].
Saputra A., Mumpuni F. S., Setiadi E., Setiawan I. D., 2019 [The growth and survival
performance of the seed of Hemibagrus nemurus catfish given different probiotics].
Jurnal Mina Sains 5(1):1-12. [In Indonesian].
Setiawati K. M., Mahardika K., Alit A. A. K., Kusumawati D., Mastuti I., 2017 [Growth and
blood profile of suni grouper (Plectropomus leopardus) seed maintained at different
salinities]. Jurnal Ilmu dan Teknologi Kelautan Tropis 9(2):557-568. [In
Indonesian].
Steel R. G. D., Torrie J. H., Dickey D. A., 1996 Principles and procedures of statistics:
Abiometrical approach. 3rd Edition. McGraw Hill, New York, pp. 352-358.
Tang U. M., 2003 [Baung fish culture technique]. Kanasius, Yogyakarta, Indonesia, 84 p.
[In Indonesian].
Tang U. M., Aryani N., Masjudi H., Hidayat K., 2018 [Effect of temperature on stress on
malay catfish (Hemibagrus nemurus)]. Asian Journal of Environment, History and
Heritage 2(1):43-49. [In Indonesian].
Uslichah U., Syandri H., 2003 [Reproduction of sasau fish (Hampala sp.) and lelan fish
(Osteochilus vittatus C.V.) in Lake Singkarak]. Jurnal Iktiologi Indonesia 1:41-48.
[In Indonesian].
Van de Nieuwegiessen P. G., Olwo J., Khong S., Verreth J. A. J., Schrama J. W., 2009
Effects of age and stocking density on the welfare of African catfish, Clarias
gariepinus Burchell. Aquaculture 288(1-2):69-75.
Wedemeyer G. A., Yasutake W. T., 1977 Clinical methods for the assessment of the
effects of environmental stress on fish health (No. 89). US Fish and Wildlife Service,
Federal Government Series, 18 p.
Wulandari Y. S., 2014 [The study on the development of African catfish (Clarias
gariepinus) cultivation using the boster system at the Fish Boster Center of Indosco

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1546
 Dwi Jaya Sakti Inc., Gedangan Subdistrict, Sidoarjo District, East Java]. UB
Repository, Malang, Indonesia, 124 p. [In Indonesian].
Yanto H., Hasan H., Sunarto, 2015 [Hematology study for early diagnosis of fish diseases
in the production center of freshwater fish culture in the Kapuas River, Pontianak
City]. Jurnal Akuatika 6(1):11-20. [In Indonesian].
Zalukhu J., Fitrani M., Sasanti A. D., 2016 [The maintenance of tilapia with different
stocking densities in aquaponic cultivation]. Jurnal Akuakultur Rawa Indonesia 4(1):
80-90. [In Indonesian].
*** Indonesian International Standard, 1990 [Collection of SNI for public works. Water
Quality SK SNI M-03-1989-F: Water quality testing methods]. Department of Public
Works, Indonesia. [In Indonesian].
*** Sudarmadji, Boster T., 2013 [S.O.P boster system catfish cultivation]. Available at:
www.indosco.com. [In Indonesian].

Received: 12 November 2020. Accepted: 19 January 2021. Published online: 06 June 2021.
Authors:
Niken Ayu Pamukas, Department of Aquaculture, Faculty of Fisheries and Marine Science, Riau University, H. R.
Subantas KM 12.5 St., 28293 Pekanbaru, Indonesia, e-mail: [email protected]
Usman Muhammad Tang, Department of Aquaculture, Faculty of Fisheries and Marine Science, Riau University,
H. R. Subantas KM 12.5 St., 28293 Pekanbaru, Indonesia, e-mail: [email protected]
Mulyadi, Department of Aquaculture, Faculty of Fisheries and Marine Science, Riau University, H. R. Subantas
KM 12.5 St., 28293 Pekanbaru, Indonesia, e-mail: [email protected]
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which
permits unrestricted use, distribution and reproduction in any medium, provided the original author and source
are credited.
How to cite this article:
Pamukas N. A., Tang U. M., Mulyadi, 2021 Hematology of Asian redtail catfish (Hemibagrus nemurus) in
different stocking densities using an aquaponic recirculation system. AACL Bioflux 14(3):1534-1547.

AACL Bioflux, 2021, Volume 14, Issue 3.

http://www.bioflux.com.ro/aacl
1547