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This research was
done to find out the lethal effects of detergent LAS on Clarias gariepinus juveniles.
The impact of exposure to linear alkyl benzene sulphonate (klin) on Clarias
gariepinus juveniles were evaluated using standard methods A total of
eighty (80) juveniles of Clarias
gariepinus were randomly
distributed into eight (8) plastic tanks for the test. They were subjected to concentrations of 0.0, 2.5, 3.3 and 4.2 mg/L
of LAS for 56 days chronic exposure study following a static renewal protocol.
The blood parameters were analyzed in the fish. The parameters like PCV and WBC
of Clarias gariepinus vary significantly (P<0.05). HB, RBC and MCH were
decreased while WBC, MCV and MCHC increased. Water quality examination showed
increase in pH, DO record was higher in the control but reduces as
concentration increases. Temperature did not differ significantly (P=0.05). Analyses of variance were
used to find out if there was a significant different in the treatment. It
was concluded that LAS causes a lot of harmful effects to fish.
Keywords: Clarias gariepinus, Hematological parameters, Linear alkyl benzene sulphonate, Water
INTRODUCTION
Water is essential natural resource of living organisms. Man depends on
water for various purposes such as fish production, drinking, cooking,
irrigation and even disposal of waste products. The aquatic environment plays
an important role in the life of all living organism due to its physicochemical
and biological properties and formation of food webs and the aquatic ecosystem.
The environment is getting polluted due to entry of different effluents into the
water bodies through domestic, agrochemicals, fertilizers, pesticides and
industrial discharges, etc. These polluting agents are toxic and deteriorate
the water quality by changing its physicochemical nature that cause an
ecological imbalance leading to stress of different kinds on aquatic organisms [1].
Detergents are organic pollutants that accumulate in freshwater
sediment, constituting pollutant mixtures [2]. One of the most common domestic
wastes that enter the aquatic ecosystem is detergent, which is a
non-biodegradable chemical substance [3].
Contamination of natural water by detergents
has become a matter of concern in recent years because of their large scale use
in home and industrial applications, such as, washing powders, dye fasteners,
formulation of shampoos, industrial and household cleansing agents, toothpaste,
tooth powder and dispersing oil spills, etc. [4].
Linear Alkyl benzene Sulphonate (LAS) is the most widely used anionic
surfactant in household and cleaning products that lower the surface tension of
water, enabling soils and stains to loosen and release from fabrics and
surfaces. These anionic surfactants are reported to be acutely toxic to aquatic
organisms [5].
Fish can serve as bio indicators of environmental pollution
(ecotoxicological studies) and therefore can be used for the assessment of the
quality of aquatic environment [6].
African catfish (Clarias gariepinus) is of great commercial
importance and it is the most common freshwater fish widely consumed in Nigeria
[7].
Clarias gariepinus
is readily recognized by their cylindrical body with scale less skin, flattened
bony head, small eyes, elongated spineless dorsal fin and four pairs of
barbells around a broad mouth. The upper surface of the head is coarsely
granulated in adult fishes but smooth in young fish [8].
MATERIALS AND METHODS
Study area
The study was carried out in the General Purpose Laboratory, Department
of Fisheries and Aquaculture, University of Agriculture, Makurdi, Benue State,
Nigeria.
Sample collection
Juveniles of Clarias gariepinus
were obtained from of the University of Agriculture, Makurdi, Benue. The fishes
were acclimatized for 14 days in the fish hatchery, Department of Fisheries and
Aquaculture, University of Agriculture, Makurdi. The fishes were fed twice
daily at 5% of their body weight during acclimatization. Prior to and during
exposure period fish were starved.
Experimental procedure
After the acclimation period, a pilot test was carried out on linear
alkyl benzene sulphonate (klin), so as to determine the lethal concentration of
LAS. Three concentrations were determined for the test at the end of the trial
which were; 2.50 mg/l, 3.30 mg/l and 4.20 mg/l.
Toxicity test
A total of one hundred and eighty (180)
juveniles of Clarias gariepinus were randomly distributed into eighteen
(18) plastic tanks. The tanks were
assigned
to 6 treatments (control inclusive) and the treatments were in triplicates. Ten fish were distributed randomly
and stocked in each tank. The exposure period lasted for 8 weeks. Fish were fed
with commercial floating feed (coppens) at 3% of their body weight.
At the end of the 8th week, blood samples of Clarias gariepinus were taken by randomly selecting fish from
the various treatments and injecting a 2 mm needle and syringe through the
dorsal aorta puncture and placed in Ethylene-Diamine-Tetra-Acetic-Acid (EDTA)
treated bottles to prevent coagulation [9]. The blood samples of Clarias gariepinus were analysed at
Laboratory College of veterinary medicine, University of agriculture Makurdi,
Benue State for the following: Hemoglobin (Hb), Packed Cell Volume (PCV), Red
Blood Cell (RBC) and White Blood Cell (WBC) while Mean Corpuscular Volume
(MCV), Mean Corpuscular Hemoglobin (MCH) and Mean Corpuscular Hemoglobin
Concentration (MCHC) using methods as described [10].
Water quality determination of test solution
The following parameters were determined during the experiment:
Temperature: Dissolved oxygen meter model HI98123 which
have ability to read both dissolved oxygen and temperature was used. The probe was immersed in the water
samples and allowed to stand for 2 min for equilibrium in each case. The
readings were taken and recorded.
Dissolved oxygen: Dissolved oxygen meter model HI98123 was used
to determine the dissolved oxygen in the water samples. The probe was dipped in
the samples of water and after waiting for about 2 min for the reading to
stabilized, the result was displayed on the LCD and the records were taken.
pH (hydrogen ion concentration):
The hydrogen ion
concentration was measured using the Hanna multi parameter water tester model H198129. The
probe was immersed in each of the water samples. It was stirred gently and
readings was taken and recorded after it has stabilized.
Total
dissolved solids: TDS
measurements were also done using Hanna HI 98129 multi parameters water
checker. The meter was set to measure the TDS by use of the MODE keypad. The
probe was immersed in each of the water samples. It was gently stirred and then
reading was taken when stabilized.
Electrical
conductivity: This was done with the use of Hanna multi parameters water checker meter.
DATA
ANALYSIS
The data obtained from these experiments were analysed using the
minitab 17th edition and one way Analysis of Variance (ANOVA).
RESULTS AND DISCUSSION
Hematological parameters of juveniles of Clarias gariepinus exposed to sub lethal concentrations of LAS are
shown in Table 1. Hematological
indices such as Packed Cell Volume (PCV) and White Blood Cell differed
significantly (P<0.01) but hemoglobin, mean corpuscular volume, mean
corpuscular hemoglobin and mean corpuscular hemoglobin concentrations were not
significantly different across the treatment. Packed Cell Volume (PCV) differed
significantly (p<0.05) with values ranging from 35.50 ± 1.00% in 0.0 mg/l to
27.00 ± 2.50 in 4.20 mg/l. White Blood Cell (WBC) also differed significantly
ranging from 11.85 ± 0.85 in 4.20 mg/l to 4.15 ± 0.65 in the control (0.0 mg/l).
Red blood cell were not significantly (P=0.99) different among the
treatments but ranged from 5.15 ± 1.35 in 0.0 mg/l to 4.65 ± 1.65 in 2.50 mg/l.
Hemoglobin value ranged from 11.85 ± 0.30
in 0.0 mg/l to 9.00 ± 0.85 in 4.20 mg/l. Mean corpuscular hemoglobin
concentration ranged from 33.30 ± 0.10 in 2.50 and 4.20 mg/l to 33.20 ± 0.10 in
0.0 mg/l, mean corpuscular hemoglobin ranged from 27.30 ± 1.04 in 2.5 mg/l to
18.55 ± 8.40 in 4.20 mg/l and mean corpuscular volume ranges from 82.00 ± 0.31
in 2.5 mg/l to 50.60 ± 2.80 in 3.3 mg/l.
Physico-chemical parameters of the water used during the experiment
Table 2 shows the physico-chemical parameters obtained during the 8 weeks sub lethal test Clarias gariepinus treated with LAS. There were significant difference (P<0.05) of the tested water among different concentrations with the exception of temperature. Dissolved oxygen decreased with increase in concentrations from 6.75 ± 0.05 mg/l in control to 4.45 ± 0.25 mg/l in 4.20 mg/l, Total dissolved solid increased with increase in concentrations also from 339.50 ± 10.50 from control to 969.00 ± 18.00 mg/l in 4.20 mg/l and so was electrical conductivity from 696.50 ± 24.50 in control to 1943.00 ± 36.00 in 4.20 mg/l and pH from 7.15 ± 0.15 in control to 8.10 ± 0.14 in 4.20 mg/l.
DISCUSSION
Blood is recognized as a potential index of fish response to water
quality, and can be used to ascertain the effects of pollutants in the environment.
Some researchers [11-15] stated that hematology may be useful tool in
monitoring stress levels of aquatic pollution on fish. Hematological parameters
are closely related to the response of the animal and to the environment, an
indication that environment where the fish lives exert some influence on the
hematological characteristics [16]. Hematology and clinical chemistry analysis,
although not used regularly, can provide substantial diagnostic information
once reference values are established. Unfortunately, reference values are not
used on a routine basis in fish and the number of studies in which reference
intervals have been determined for fish is limited [17]. However, it is well
known that blood sampling, laboratory techniques, seasoned variation, size,
genetic properties, sex, population density, lack of food supply, environmental
stress and transportation could affect hematological data [18]. Hence,
comparison of reference interval should be done with caution in respect to
variation in environmental condition.
Hematological parameters of Clarias
gariepinus such as PCV, HB, RBC
and MCH showed reduction with increase in concentration while WBC, MCV and MCHC
increased with increased LAS concentrations. This situation is similar to that
reported by some researchers [19-21] on T. guinensis. Low PCV value
obtained in this study may be attributed to reduction in red blood cell caused
by osmotic changes [22]. This finding is in agreement with the findings of others
who investigated hematological changes in mudfish Clarias gariepinus exposed to sub lethal concentrations of copper
and lead [23,24]. The observed decrease in the PCV and Hb of C. gariepinus
juvenile exposed to LAS could be indicative of hemodilution due to
erythrocyte sequestration. Some workers have attributed changes in such blood
parameters to erythrocyte swelling [25] or hemolysis [26]. Some have reported
that the RBC count decreased significantly in the fresh water fish, Labeo
rohita on exposure to herbicide glyphosate [27,28]. While others reported
decreased hemoglobin content in the fresh water fish, Tilapia mossambica on
exposure to arsenic [29,30]. Certain researchers made similar observation in Cirrhinus
mrigala exposed to detergent tide [31,32]. All these observations are in
conformity with the findings of the present study. Thus, a reduced red blood
cell count implies a reduction in the level of oxygen that would be carried to
the tissues as well as the level of carbon dioxide returned to the lungs [33].
Although the increasing or decreasing number of the White Blood Cell
(WBC) of Clarias gariepinus is a
normal reaction on the exposure of fish to toxicants [34], this study showed
increase with increased concentration of LAS to Clarias gariepinus. This is in agreement with the work of certain
scientists who reported increased WBC with increase in concentration of tobacco
leaf dust. This may be attributed to increase in leucocytes synthesis as a
defense mechanism against the destruction of erythrocytes [35].
MCH and MCHC is an indicator of red blood cell swelling. In the
findings, MCH of Clarias gariepinus
reduced with increase in concentrations of LAS where as MCHC and MCV increased
with increase in concentration of LAS in Clarias
gariepinus. This is in consonant to the findings of which reported a
decrease in MCH and increase in MCV [36]. The increase in MCV is contrary to
the findings of some of the researchers who reported lower MCV values in Clarias gariepinus treated with Zinc
salt [37].
Effects on water quality parameters
The temperature reading for acute and sub lethal effect of LAS on Clarias
gariepinus fell within 23-30°C in the study which is in accordance with
the range (20-30°C) suggested by the Federal Environment Protection Agency for
optimum physiological state of fish. Dissolved oxygen reduced with increase in
concentrations of with control having the highest dissolved oxygen in both
acute and sub lethal test and lowest value was recorded in the highest
concentration for acute and sub lethal test. The heighten activities of the
fish due to poison can also remove oxygen from the water body had earlier
reported that indiscriminate deposition of effluent into an aquatic system
might decrease the dissolved oxygen concentration, which stand to impair
respiration leading to asphyxiation (which is an indication of unconsciousness
or death produced by failure of the blood to become properly oxygenated in the
lungs) and may ultimately result into organ architectural degradation such as
liver dysfunction. Decrease in
dissolved oxygen in this study can be attributed to the presence of the
chemical (LAS) in the water. TDS, EC and pH increased with increase in
concentrations. The water quality
parameters of the study were significantly different from the control. However,
they were within acceptable tolerance range and also could not have had any
negative effect on the tested fish. The water quality parameters showed
significant difference (p<0.05) with exception of the Temperature both in
the acute and sub lethal of Clarias
gariepinus on LAS.
A parameter like temperature can influence metabolic rate in the bodies
of organisms affect density of ambient water of the organism and even food
availability [15]. In the present experiment, temperatures did not vary both
acute and sub lethal test of Oreochromis
niloticus but was between 26 and 28°C which are within tolerance limits for
the survival of fish [16]. Some recommended a pH range of 6.5-9. The pH of both
acute and sub lethal test of Oreochromis
niloticus were also within the tolerance limit of fish survival, it could
not have affected the fish, though slight changes in pH could grossly affect
ammonium nitrogen toxicity [38]. It was affirmed that detergent concentration
as little as 0.9 mg/l at a pH of 5.5 is lethal to fish. Significant variations
in oxygen concentrations were observed, though within tolerance limit, reduced
levels as observed with increased LAS concentration could have led to stress.
CONCLUSION
1. In
conclusion, linear alkyl benzene sulphonate was found to be toxic to juveniles
of Clarias gariepinus.
2. Histopathological
studies further revealed that acute concentrations of LAS damaged the gills and
liver of treated fishes and led to their deaths.
3. Sub
lethal concentrations of LAS adversely affected the growth performances of C. gariepinus and Oreochromis niloticus.
4. The
present study revealed that sub lethal concentrations of LAS in C. gariepinus and Oreochromis niloticus created hematological disturbances.
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