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International Journal of Trend in Scientific Research and Development (IJTSRD)
Volume 4 Issue 2, February 2020 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470
@ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 279
Bioaccumulation of Heavy Metals in
Clarias Gariepinus Exposed to Untreated Paint Effluent
Orimoloye, M. A; Mohammed, A
Department of Science Laboratory Technology, Rufus Giwa Polytechnic, Owo, Nigeria
ABSTRACT
This study presents the heavy metal accumulationprofileinClariasgariepinus
(catfish) exposed to untreated paint effluent. Sub-lethal concentrations of
paint effluent were used to pollute catfish aquaria. This was followed by
isolation and identification of microorganisms from the pollutedaquaria after
96 h using conventional microbiological techniques. The tissue heavy metal
composition and physicochemical parameters of the aquaria were also
monitored using standard analytical methods. Microbial load in the range of
10.00a±2.00 to 189.00d±20.52 cfu/ml ontissuesofcatfishwereobserved.The
heavy metals present in the paint effluent were mainly Lead (1.20±0.01ppm)
and Zinc (0.34±0.01ppm). No significant difference in the proximate analysis
between the tissue of catfish from the control and the polluted aquaria were
observed. Catfish tissue accumulated Lead (0.16 ppm) from the polluted
aquaria. Persistent exposure through pollution by effluent in aquatic
environment can lead to increased metallic Lead bioaccumulation that could
be unhealthy for both the fish and human consumption. Therefore,
indiscriminate discharge of untreated effluent from industrial sitesinto water
bodies should be discouraged.
KEYWORDS: Clarias gariepinus, bioaccumulation, heavy metal, paint effluent
How to cite this paper: Orimoloye, M. A |
Mohammed, A "BioaccumulationofHeavy
Metals in Clarias Gariepinus Exposed to
Untreated Paint
Effluent" Published
in International
Journal of Trend in
Scientific Research
and Development
(ijtsrd), ISSN: 2456-
6470, Volume-4 |
Issue-2, February 2020, pp.279-284,URL:
www.ijtsrd.com/papers/ijtsrd29989.pdf
Copyright © 2019 by author(s) and
International Journal ofTrendinScientific
Research and Development Journal. This
is an Open Access article distributed
under the terms of
the Creative
CommonsAttribution
License (CC BY 4.0)
(http://creativecommons.org/licenses/by
/4.0)
1. INTRODUCTION
Persistent contamination of water bodies through continuous discharge of industrial effluents is common and can produce
varying degree of physiology and biochemical changes in fish species [1]. Rapid urbanization and industrialization has led to
increased disposal of pollutants such as heavy metals,radionuclidesandvarioustypes oforganicandinorganicinto theaquatic
environment. Extensive industrialization has measurably influences the quality of water of lakes, pondsandrivers all overthe
world. Industrial wastes constitute the majorsourceof metal contaminationoftheaquaticenvironment[2].Industrial effluents
have been regarded as the main input source of various pollutants to the aquatic ecosystem and contain various organic and
inorganic substances potentially toxic to aquatic biota. It is well established that pollution lowers the quality of life in various
aspects and affects health and life span [3]. Besides the direct health effects, the subtle danger of pollutants lies in the fact that
they may be mutagenic or toxic and lead to several human afflictions like cancer, atherosclerosis, cardiovascular diseasesand
premature ageing [4]. Discharging effluents without proper treatments may have an adverse effect on the receiving water
bodies [5]. Heavy metals constitute the major contaminants. Metals are important pollutants, because they are not eliminated
from the aquatic ecosystems by natural processes like organic pollutants and are enriched in mineral organic substances.
Metallic contaminants are introduced into aquatic system through smelting process, effluents,sewageandleaching ofgarbage
which cause serious damage to the aquatic fauna [6]. Heavy metals have long being recognized as serious pollutants of the
aquatic ecosystem. Increase discharge of heavy metals into natural aquatic ecosystems can expose aquatic organisms to
unnatural high levels of these metals [7]. The metals entering the aquatic ecosystem may not directly cause damage to
organism but they can be deposited in aquatic organism through the effect of bio-concentration, bioaccumulation and other
food chain processes thus gets to man in concentrations that poses threats to human health via consumption of sea foods [8].
Transport of metals in fish occurs through the blood where the ions are usually bound to proteins.Themetalsarebroughtinto
contact with the organs and tissue of the fish and consequently accumulated to a different extent in different organs ortissues
of the fish. Once heavy metals are accumulated by an aquatic organism, they can be transported through the upper classofthe
food chain [9].
Fish is an important component of human nutrition, and those from contaminated sites present a potential risk to human
health. Metals can accumulate in aquatic organisms includingfishandpersistinwaterandsediments.Fishesarethe simpleand
reliable biomarkers of pollution of metal in a polluted aquatic ecosystem, they tend to take these metals up from their direct
environment [10].Therefore, this study was undertaken to assess the heavy metal accumulationandmicrobial safetyofcatfish
grown in aquaria contaminated with paint effluent.
IJTSRD29989
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 280
2. MATERIALS AND METHODS
2.1. Collection of samples
The Paint effluent used in this study was obtained at point source of a Paint Factory in Lagos. The samples were collected in
sterile containers and taken for physiochemical and microbiological analysis.
2.2. Set up and Pollution of Aquaria
Six (6) aquaria in duplicates each contain 10 fingerlings. The fishes were then exposed to paint effluent at sub-lethal
concentrations of 0.00 (control), 1.25mls/l, 2.50mls/l, 3.75mls/l, 5.0mls/l and 6.25mls/l for 96hours. Careful observations
were then made to note the number of mortalities and effect of the paint on the test organism.
Analytical Methods
2.3. Water Analysis for Heavy Metal
Water samples in the bottles were shaken sufficiently for thorough mixing, and then 50mls aliquots were filtered on Watman
No. 41 filter papers and quantitatively transferred into 50mls volumetric flask before aspiring directly into the Inductively
Coupled Plasms for analysis of heavy metals as describe by APHA [11].
2.4. Analysis on fish
2.4.1. Heavy Metals Analysis
The concentrations of the selected heavy metals (Pb and Zn) in fish flesh were determined using X-Ray Fluorescenceanalyzer.
2.4.2. Nutritive Value Analysis of Fish
Nutritive value of the sample flesh was determined using Kjedahl Steam Distillation and Soxhlet methods respectively.
2.5. Physiochemical Parameters
The physiochemical parameters suchastemperature,total dissolvedsolidsandtotal solidsweredeterminedusingthemethods
of Aderinola et al. [12].
2.6. Biochemical and Morphological Identification of Bacteria Isolates
Bacteria were isolated using nutrient agar as well as selective media; and the individual colonies were identified by
morphological and biochemical techniques using methods described by Osho and Fawole [13].
2.7. Statistical Analysis
The data were analyzed for significantdifferences(P<0.05)byAnalysisofVariance(ANOVA)usingcomputerStatistical Package
for Social Sciences (SPSS) for windows (v.25.0). Determined differences were partitioned by the Least Significant Difference
(LSD) at P=0.05
3. Results and discussion
The temperature range of the aquaria contaminated with paint effluent is presentedin figure1.Thefigureshowsnosignificant
difference in the temperature of the control and other treatmentsexcepttreatment2whichhada lowertemperatureatthe end
of the experimental period. This suggest that the paint effluents had no effect on the temperature of the fish aquaria however,
the lower temperature observed in the treatment 2 may need to be investigated more since there are no observationthatmay
suggest possible explanation for the drop in the temperature.
Fig1: Temperature of water in catfish aquaria
Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 281
Fig 2: Total dissolved solids in catfish water aquaria
Fig 3: Total solids in catfish water aquaria
Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L
Figures 2 and 3 presents the total dissolved solids and total solids in the aquaria water samplesrespectively.Thetotal solidsin
the aquaria water increased with increase in the paint effluent contamination rising from 0.125mg/L in the control to
0.265mg/L in the treatment 5 (6.25ml/L of effluent)atthebeginningoftheexperiment.Meanwhile,thetotal solidsgotreduced
drastically in all the treated samples after four days trial except treatment 1. This may be due to assimilation of some of the
solid particles by the fish [14]. Also, it may be due to settling of these solids at the bottom of the aquaria
Table1: Heavy metal present in paint effluent sample
Parameters Composition
Zinc 0.34±0.01
Nickel N.D
Chromium N.D
Cadmium N.D
Lead 1.20±0.01
Key: ND- Not detected
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 282
Table2: Proximate composition of catfish
Treatment Moisture content Crude protein Fat contents Ash content
Control 7.78c±0.12 32.71a±0.75 27.62a±0.38 11.73b±0.20
TR1 4.92a±0.21 47.86c±0.87 28.33a±0.23 13.77c±0.40
TR2 6.54b±0.48 38.88b±0.00 29.52a±0.13 10.50a±0.40
TR3 10.02d±0.07 48.5c±0.00 31.54ab±0.02 11.30ab±0.06
TR4 8.68c±0.45 31.71a±0.78 29.01a±0.05 10.46a±0.27
TR5 7.66c±0.29 37.17b±0.78 30.67ab±0.32 11.03ab±0.11
Values are presented as Mean±S.E (n=3). Means with the same superscript letter(s) along the same column are significantly
different (P>0.05).
Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L
Table3: Heavy metal accumulation in harvested catfish
Treatments Zinc Lead
Control 0.21 0.01
TR1 0.22 0.06
TR2 0.24 0.09
TR3 0.15 0.12
TR4 0.14 0.16
TR5 0.22 0.15
Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L
Table4: Bacterial load of catfish tissue (x 102 cfu/g)
Treatment Skin Liver Gill Intestine
Control 10.00a±2.00 33.67a±3.67 35.33a±7.57 100.67b±1.53
TR1 58.67c±13.32 75.67ab±4.30 80.67b±21.20 142.00c±5.05
TR2 70.67b±10.07 87.67bc±8.02 81.33b±6.11 180.00e±4.00
TR3 103.67c±4.73 126.33c±5.51 91.33d±3.22 104.67b±2.01
TR4 100.33c±20.50 90.00bc±15.10 89.00d±20.52 85.67a±6.03
TR5 73.00b±20.66 77.33ab±8.08 85.67c±16.01 168.00d±5.92
Values are presented as Mean±S.E (n=3). Means with the same superscript letter (s) along the same column are not
significantly different (P>0.05).
Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L
Table5: Frequency of occurrences of bacterial isolates in catfish grown in paint effluent contaminated aquaria
Isolates Before After Total
Staphylococcus aureus 2 1 3
Salmonella sp _ 2 2
Bacillus substilis 3 4 7
Enterobacter aerogenes 2 _ 2
Bacillus cereus 2 3 5
Micrococcus leteus 1 2 3
Klebsiella sp 1 1 2
Aeromonas hydrophila 2 1 3
Listeria monocytogenes _ 2 2
Shigella sp 1 1 2
The results of the heavy metal present in the painteffluentis
shown in table 1 where it was revealed that Ni, Cr, and Cd
were not detected in the effluent whereas, Pb and Zn were
found to be 1.20 ppm and 0.34 ppm respectively. The non
detection of the other heavy metals in the effluent samples
may be due to pretreatment bythecompanybefore releasing
their waste water into the environment, since the
government require industries to treat their wastes to
reduce toxicity to barest minimum before ejectingsameinto
the environment [15]. The presence of lead in the effluent in
high proportion suggests that the effluentmaybedangerous
to aquatic lives if released like that in to water bodies. Lead
poisoning is one of the major problems associated with
industrial effluent discharge with debilitating effects on
animals. The heavy metals become toxic when they do not
get metabolized by the body and end up accumulating in the
soft tissues [16]. Ingestion is the most common route of
exposure to heavy metals. Fish has been reported to
accumulate metals from water by diffusion via skin and gills
as well as oral consumption. Bioaccumulation of heavy
metals in Clarias gariepinus probably might have resulted
from inability of the fish to metabolize the heavy metals or
the metabolism of the heavy metals in the fish tissues, were
slow [17].
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 283
The results of the proximate composition and mineral
content of the catfish used in this study are presented in
table 2 and 3 respectively. There were no significant
differences in the crude protein, fat and ash content of the
fish. However, there was a gradual increase in lead
concentration in the tissue of the cat fish withincreaseinthe
concentration of the paint effluent. The increase may be due
to the availability of the metal in the water where the fish
lives as a result of contamination by the effluent. Also,
bioaccumulation of heavy metals especially lead in aquatic
animals have been documented in the last decade. The high
level of lead in the catfish tissue may be linked with
bioaccumulation of same. Similar result was also
documented by Igwegbe et al. [18].
The bacterial load on different parts of the cat fish is shown
in table 4. The table revealed that all the catfish grown in
paint effluent contaminated aquaria had higher bacterial
count compared with the control. The bacterial load of the
intestines were higher than those found in other parts ofthe
body whereas the skin had the least count. The higher count
in the intestine may be due to digestion activity going on in
the intestine as well as the abundance of food available for
microbial growth [19]. However, the high microbial count
ranging from 75.67 to 126./33 x 102 cfu/g observed in the
liver compared with 33.67 x 102 cfu/g obtained in the
control group suggests that the paint effluent may suppress
the catfish immunity and allow infection of major internal
organs. This may lead to death of the fish and general loss of
aquatic diversity [20]. A total of 10 bacterial isolates were
obtained based on their morphological and biochemical
properties. The bacteria were: Micrococcus luteus, Bacillus
subtilis, Staphylococcus aureus. Enterobacter aerogenes,
Salmonella Sp, Bacillus cereus, Klebsiella spp, Aeromonas
hydrophila, and Listeria monocytogenes. Tables 3 shows the
frequency of occurrences of probable isolatesfromcatfish,it
was observed that Bacillus subtilis was the most frequentfor
catfish. Some of these bacteria are known pathogens of man
and their presence in the fish suggests that the consumption
of the fish pose serious health risk to the consumers [21].
4. Conclusion
This study revealed the quality of fish in terms of heavy
metal and microbial contamination, when exposed to an
industrial paint effluent. The levels of the heavy metals
detected in fish are not remarkable to cause acute health
consequences, but chronic adverse health effects due to
prolonged bioaccumulation andlongterm exposurethrough
fish consumption can be inevitable. The public should be
enlightened on the possibility of being susceptible to health
problems when feeding on fish contaminated with heavy
metals and harmful microorganisms
References
[1] J. A. Adakole and D. S. Abolude, “Studies on effluent
characteristics of a metal finishing company, Zaria –
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[8] J. G. Holt, N. R. Krieg, P. H. A. Sneath, J. T. Stanley and S.
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[9] R. Karbassi, I. Bayati and J. Moattar. “Origin and
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[10] E. H. Kim, Y. R. Jun, H. J. Jo, S. B. Shim and J. H. Jung.
“Toxicity identification in metal plating effluent:
implications in establishing effluent discharge limits
using bioassays in Korea”. Mar. Pollut. Bull. Vol 57, pp
637–644, 2008.
[11] American Public HealthAssociation.StandardMethods
for the Examinationof Water andWastewaterAnalysis,
(21st ed. ). American Water Works Association/Water
Environment Federation, Washington D. C, pp. 289,
2005.
[12] O. J. Aderinola, V. Kusemiju and E. O. Clarke. “Trace
metal distribution in surface water, sediment, and
tissues of freshwater catfish (Clarias. gariepinus),from
Oke- Afa canal, Lagos Nigeria. ” International Journal of
Geography and Geology, VOL. 1(1), PP 10-22, 2012.
[13] O. M. Fawole and B. A. Oso. “Laboratory manual of
Microbiology. ” 5th Edition, Spectrum Books Limited,
Ibadan, PP 22-23, 2007.
[14] D. R. Livingstone. “Oxidative stress inaquaticorganism
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[15] J. C. Van Dyk, G. M. Pieterse and J. H. J. Van Vuren.
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[16] S. Hayat and M. Javed. “Regression studies of
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@ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 284
with sub-lethal metal stressed fish. ” International
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[17] J. C. Akan, M. Salwa B. S. Yikala and Z. M Chellube.
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[18] A. O. Igwegbe, A. N. Charles, C. C. Elizabeth and H. B.
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[20] G. Nussey, J. H. J. Van Vuren, and H. H. Du Preez.
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Bioaccumulation of Heavy Metals in Clarias Gariepinus Exposed to Untreated Paint Effluent

  • 1. International Journal of Trend in Scientific Research and Development (IJTSRD) Volume 4 Issue 2, February 2020 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470 @ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 279 Bioaccumulation of Heavy Metals in Clarias Gariepinus Exposed to Untreated Paint Effluent Orimoloye, M. A; Mohammed, A Department of Science Laboratory Technology, Rufus Giwa Polytechnic, Owo, Nigeria ABSTRACT This study presents the heavy metal accumulationprofileinClariasgariepinus (catfish) exposed to untreated paint effluent. Sub-lethal concentrations of paint effluent were used to pollute catfish aquaria. This was followed by isolation and identification of microorganisms from the pollutedaquaria after 96 h using conventional microbiological techniques. The tissue heavy metal composition and physicochemical parameters of the aquaria were also monitored using standard analytical methods. Microbial load in the range of 10.00a±2.00 to 189.00d±20.52 cfu/ml ontissuesofcatfishwereobserved.The heavy metals present in the paint effluent were mainly Lead (1.20±0.01ppm) and Zinc (0.34±0.01ppm). No significant difference in the proximate analysis between the tissue of catfish from the control and the polluted aquaria were observed. Catfish tissue accumulated Lead (0.16 ppm) from the polluted aquaria. Persistent exposure through pollution by effluent in aquatic environment can lead to increased metallic Lead bioaccumulation that could be unhealthy for both the fish and human consumption. Therefore, indiscriminate discharge of untreated effluent from industrial sitesinto water bodies should be discouraged. KEYWORDS: Clarias gariepinus, bioaccumulation, heavy metal, paint effluent How to cite this paper: Orimoloye, M. A | Mohammed, A "BioaccumulationofHeavy Metals in Clarias Gariepinus Exposed to Untreated Paint Effluent" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456- 6470, Volume-4 | Issue-2, February 2020, pp.279-284,URL: www.ijtsrd.com/papers/ijtsrd29989.pdf Copyright © 2019 by author(s) and International Journal ofTrendinScientific Research and Development Journal. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (CC BY 4.0) (http://creativecommons.org/licenses/by /4.0) 1. INTRODUCTION Persistent contamination of water bodies through continuous discharge of industrial effluents is common and can produce varying degree of physiology and biochemical changes in fish species [1]. Rapid urbanization and industrialization has led to increased disposal of pollutants such as heavy metals,radionuclidesandvarioustypes oforganicandinorganicinto theaquatic environment. Extensive industrialization has measurably influences the quality of water of lakes, pondsandrivers all overthe world. Industrial wastes constitute the majorsourceof metal contaminationoftheaquaticenvironment[2].Industrial effluents have been regarded as the main input source of various pollutants to the aquatic ecosystem and contain various organic and inorganic substances potentially toxic to aquatic biota. It is well established that pollution lowers the quality of life in various aspects and affects health and life span [3]. Besides the direct health effects, the subtle danger of pollutants lies in the fact that they may be mutagenic or toxic and lead to several human afflictions like cancer, atherosclerosis, cardiovascular diseasesand premature ageing [4]. Discharging effluents without proper treatments may have an adverse effect on the receiving water bodies [5]. Heavy metals constitute the major contaminants. Metals are important pollutants, because they are not eliminated from the aquatic ecosystems by natural processes like organic pollutants and are enriched in mineral organic substances. Metallic contaminants are introduced into aquatic system through smelting process, effluents,sewageandleaching ofgarbage which cause serious damage to the aquatic fauna [6]. Heavy metals have long being recognized as serious pollutants of the aquatic ecosystem. Increase discharge of heavy metals into natural aquatic ecosystems can expose aquatic organisms to unnatural high levels of these metals [7]. The metals entering the aquatic ecosystem may not directly cause damage to organism but they can be deposited in aquatic organism through the effect of bio-concentration, bioaccumulation and other food chain processes thus gets to man in concentrations that poses threats to human health via consumption of sea foods [8]. Transport of metals in fish occurs through the blood where the ions are usually bound to proteins.Themetalsarebroughtinto contact with the organs and tissue of the fish and consequently accumulated to a different extent in different organs ortissues of the fish. Once heavy metals are accumulated by an aquatic organism, they can be transported through the upper classofthe food chain [9]. Fish is an important component of human nutrition, and those from contaminated sites present a potential risk to human health. Metals can accumulate in aquatic organisms includingfishandpersistinwaterandsediments.Fishesarethe simpleand reliable biomarkers of pollution of metal in a polluted aquatic ecosystem, they tend to take these metals up from their direct environment [10].Therefore, this study was undertaken to assess the heavy metal accumulationandmicrobial safetyofcatfish grown in aquaria contaminated with paint effluent. IJTSRD29989
  • 2. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 280 2. MATERIALS AND METHODS 2.1. Collection of samples The Paint effluent used in this study was obtained at point source of a Paint Factory in Lagos. The samples were collected in sterile containers and taken for physiochemical and microbiological analysis. 2.2. Set up and Pollution of Aquaria Six (6) aquaria in duplicates each contain 10 fingerlings. The fishes were then exposed to paint effluent at sub-lethal concentrations of 0.00 (control), 1.25mls/l, 2.50mls/l, 3.75mls/l, 5.0mls/l and 6.25mls/l for 96hours. Careful observations were then made to note the number of mortalities and effect of the paint on the test organism. Analytical Methods 2.3. Water Analysis for Heavy Metal Water samples in the bottles were shaken sufficiently for thorough mixing, and then 50mls aliquots were filtered on Watman No. 41 filter papers and quantitatively transferred into 50mls volumetric flask before aspiring directly into the Inductively Coupled Plasms for analysis of heavy metals as describe by APHA [11]. 2.4. Analysis on fish 2.4.1. Heavy Metals Analysis The concentrations of the selected heavy metals (Pb and Zn) in fish flesh were determined using X-Ray Fluorescenceanalyzer. 2.4.2. Nutritive Value Analysis of Fish Nutritive value of the sample flesh was determined using Kjedahl Steam Distillation and Soxhlet methods respectively. 2.5. Physiochemical Parameters The physiochemical parameters suchastemperature,total dissolvedsolidsandtotal solidsweredeterminedusingthemethods of Aderinola et al. [12]. 2.6. Biochemical and Morphological Identification of Bacteria Isolates Bacteria were isolated using nutrient agar as well as selective media; and the individual colonies were identified by morphological and biochemical techniques using methods described by Osho and Fawole [13]. 2.7. Statistical Analysis The data were analyzed for significantdifferences(P<0.05)byAnalysisofVariance(ANOVA)usingcomputerStatistical Package for Social Sciences (SPSS) for windows (v.25.0). Determined differences were partitioned by the Least Significant Difference (LSD) at P=0.05 3. Results and discussion The temperature range of the aquaria contaminated with paint effluent is presentedin figure1.Thefigureshowsnosignificant difference in the temperature of the control and other treatmentsexcepttreatment2whichhada lowertemperatureatthe end of the experimental period. This suggest that the paint effluents had no effect on the temperature of the fish aquaria however, the lower temperature observed in the treatment 2 may need to be investigated more since there are no observationthatmay suggest possible explanation for the drop in the temperature. Fig1: Temperature of water in catfish aquaria Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L
  • 3. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 281 Fig 2: Total dissolved solids in catfish water aquaria Fig 3: Total solids in catfish water aquaria Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L Figures 2 and 3 presents the total dissolved solids and total solids in the aquaria water samplesrespectively.Thetotal solidsin the aquaria water increased with increase in the paint effluent contamination rising from 0.125mg/L in the control to 0.265mg/L in the treatment 5 (6.25ml/L of effluent)atthebeginningoftheexperiment.Meanwhile,thetotal solidsgotreduced drastically in all the treated samples after four days trial except treatment 1. This may be due to assimilation of some of the solid particles by the fish [14]. Also, it may be due to settling of these solids at the bottom of the aquaria Table1: Heavy metal present in paint effluent sample Parameters Composition Zinc 0.34±0.01 Nickel N.D Chromium N.D Cadmium N.D Lead 1.20±0.01 Key: ND- Not detected
  • 4. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 282 Table2: Proximate composition of catfish Treatment Moisture content Crude protein Fat contents Ash content Control 7.78c±0.12 32.71a±0.75 27.62a±0.38 11.73b±0.20 TR1 4.92a±0.21 47.86c±0.87 28.33a±0.23 13.77c±0.40 TR2 6.54b±0.48 38.88b±0.00 29.52a±0.13 10.50a±0.40 TR3 10.02d±0.07 48.5c±0.00 31.54ab±0.02 11.30ab±0.06 TR4 8.68c±0.45 31.71a±0.78 29.01a±0.05 10.46a±0.27 TR5 7.66c±0.29 37.17b±0.78 30.67ab±0.32 11.03ab±0.11 Values are presented as Mean±S.E (n=3). Means with the same superscript letter(s) along the same column are significantly different (P>0.05). Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L Table3: Heavy metal accumulation in harvested catfish Treatments Zinc Lead Control 0.21 0.01 TR1 0.22 0.06 TR2 0.24 0.09 TR3 0.15 0.12 TR4 0.14 0.16 TR5 0.22 0.15 Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L Table4: Bacterial load of catfish tissue (x 102 cfu/g) Treatment Skin Liver Gill Intestine Control 10.00a±2.00 33.67a±3.67 35.33a±7.57 100.67b±1.53 TR1 58.67c±13.32 75.67ab±4.30 80.67b±21.20 142.00c±5.05 TR2 70.67b±10.07 87.67bc±8.02 81.33b±6.11 180.00e±4.00 TR3 103.67c±4.73 126.33c±5.51 91.33d±3.22 104.67b±2.01 TR4 100.33c±20.50 90.00bc±15.10 89.00d±20.52 85.67a±6.03 TR5 73.00b±20.66 77.33ab±8.08 85.67c±16.01 168.00d±5.92 Values are presented as Mean±S.E (n=3). Means with the same superscript letter (s) along the same column are not significantly different (P>0.05). Key: TR1; 1.25mls/L, TR2; 2.50mls/L, TR3; 3.75mls/L, TR4; 5.0mls/L TR5; 6.25mls/L Table5: Frequency of occurrences of bacterial isolates in catfish grown in paint effluent contaminated aquaria Isolates Before After Total Staphylococcus aureus 2 1 3 Salmonella sp _ 2 2 Bacillus substilis 3 4 7 Enterobacter aerogenes 2 _ 2 Bacillus cereus 2 3 5 Micrococcus leteus 1 2 3 Klebsiella sp 1 1 2 Aeromonas hydrophila 2 1 3 Listeria monocytogenes _ 2 2 Shigella sp 1 1 2 The results of the heavy metal present in the painteffluentis shown in table 1 where it was revealed that Ni, Cr, and Cd were not detected in the effluent whereas, Pb and Zn were found to be 1.20 ppm and 0.34 ppm respectively. The non detection of the other heavy metals in the effluent samples may be due to pretreatment bythecompanybefore releasing their waste water into the environment, since the government require industries to treat their wastes to reduce toxicity to barest minimum before ejectingsameinto the environment [15]. The presence of lead in the effluent in high proportion suggests that the effluentmaybedangerous to aquatic lives if released like that in to water bodies. Lead poisoning is one of the major problems associated with industrial effluent discharge with debilitating effects on animals. The heavy metals become toxic when they do not get metabolized by the body and end up accumulating in the soft tissues [16]. Ingestion is the most common route of exposure to heavy metals. Fish has been reported to accumulate metals from water by diffusion via skin and gills as well as oral consumption. Bioaccumulation of heavy metals in Clarias gariepinus probably might have resulted from inability of the fish to metabolize the heavy metals or the metabolism of the heavy metals in the fish tissues, were slow [17].
  • 5. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 283 The results of the proximate composition and mineral content of the catfish used in this study are presented in table 2 and 3 respectively. There were no significant differences in the crude protein, fat and ash content of the fish. However, there was a gradual increase in lead concentration in the tissue of the cat fish withincreaseinthe concentration of the paint effluent. The increase may be due to the availability of the metal in the water where the fish lives as a result of contamination by the effluent. Also, bioaccumulation of heavy metals especially lead in aquatic animals have been documented in the last decade. The high level of lead in the catfish tissue may be linked with bioaccumulation of same. Similar result was also documented by Igwegbe et al. [18]. The bacterial load on different parts of the cat fish is shown in table 4. The table revealed that all the catfish grown in paint effluent contaminated aquaria had higher bacterial count compared with the control. The bacterial load of the intestines were higher than those found in other parts ofthe body whereas the skin had the least count. The higher count in the intestine may be due to digestion activity going on in the intestine as well as the abundance of food available for microbial growth [19]. However, the high microbial count ranging from 75.67 to 126./33 x 102 cfu/g observed in the liver compared with 33.67 x 102 cfu/g obtained in the control group suggests that the paint effluent may suppress the catfish immunity and allow infection of major internal organs. This may lead to death of the fish and general loss of aquatic diversity [20]. A total of 10 bacterial isolates were obtained based on their morphological and biochemical properties. The bacteria were: Micrococcus luteus, Bacillus subtilis, Staphylococcus aureus. Enterobacter aerogenes, Salmonella Sp, Bacillus cereus, Klebsiella spp, Aeromonas hydrophila, and Listeria monocytogenes. Tables 3 shows the frequency of occurrences of probable isolatesfromcatfish,it was observed that Bacillus subtilis was the most frequentfor catfish. Some of these bacteria are known pathogens of man and their presence in the fish suggests that the consumption of the fish pose serious health risk to the consumers [21]. 4. Conclusion This study revealed the quality of fish in terms of heavy metal and microbial contamination, when exposed to an industrial paint effluent. The levels of the heavy metals detected in fish are not remarkable to cause acute health consequences, but chronic adverse health effects due to prolonged bioaccumulation andlongterm exposurethrough fish consumption can be inevitable. The public should be enlightened on the possibility of being susceptible to health problems when feeding on fish contaminated with heavy metals and harmful microorganisms References [1] J. A. Adakole and D. S. Abolude, “Studies on effluent characteristics of a metal finishing company, Zaria – Nigeria”, Res. J. Environ. Res. , vol 1(2) PP 54-57, 2009. [2] S. O. Adewoye and O. O. Fawole. “Bioconcentration of metals in the tissue of Clarias gariepinus fingerlings exposed to lethal concentration of cassava Waste water. ” World J. Biotechnol. , vol 3, PP 465-468, 2002 [3] M. Al – Masri, S. Aba, A. H. Khalil and Z. Al – Hares. “Sedimentation rates and pollution history of a dried lake” Sci. Total Environ. , vol 293 (1–3), pp 177 – 189, 2002. [4] T. A. Ayandiran, O. O. Fawole, S. O. Adewoye and M. A. Ogundiran. “Bioconcentration of metals in the body muscle and gut of Clarias gariepinus exposed to sublethal concentrations of soap and detergent effluent.” J. Cell Anim. Biol. , vol. 3(8), pp 113-118, 2009. [5] Y. K. Chan, C. K. Wong, D. P. H. Hsieh, S. P. Ng, T. K. Lau, and P. K. Wong. “Application of a toxicity identification evaluation for a sample of effluent discharged from a dyeing factory in Hong Kong. ” Environ. Toxicol. 18, PP 312–316, 2003. [6] C. E. Gomez, L. Contento and A. E. Carsen. “Toxicity tests to assess pollutants removal during wastewater treatment and the quality of receiving waters in Argentina.” Environ. Toxicol. 16, pp 217–224, 2001 [7] U. U. Gabriel, F. G. Obomanu and O. S. Edori. “Biochemical changes in hybrid cat fish (Heterobrartchus bidorsalis, Clarias gariepinus). Treated with Nuracron. ” Chinese J. AppL Environ.Biol. VOL 16 (3): PP 353-357, 2010. [8] J. G. Holt, N. R. Krieg, P. H. A. Sneath, J. T. Stanley and S. T. William. “Bergey’s Manual Of Determinative Bacteriology.” Williams and Wilikins, Baltimore, pp 786-788, 1994. [9] R. Karbassi, I. Bayati and J. Moattar. “Origin and chemical partioning of heavy metals in riverbed sediments” Int. J. Environ. Sci. Tech. , vol. 3(1) pp 35 – 42, 2006. [10] E. H. Kim, Y. R. Jun, H. J. Jo, S. B. Shim and J. H. Jung. “Toxicity identification in metal plating effluent: implications in establishing effluent discharge limits using bioassays in Korea”. Mar. Pollut. Bull. Vol 57, pp 637–644, 2008. [11] American Public HealthAssociation.StandardMethods for the Examinationof Water andWastewaterAnalysis, (21st ed. ). American Water Works Association/Water Environment Federation, Washington D. C, pp. 289, 2005. [12] O. J. Aderinola, V. Kusemiju and E. O. Clarke. “Trace metal distribution in surface water, sediment, and tissues of freshwater catfish (Clarias. gariepinus),from Oke- Afa canal, Lagos Nigeria. ” International Journal of Geography and Geology, VOL. 1(1), PP 10-22, 2012. [13] O. M. Fawole and B. A. Oso. “Laboratory manual of Microbiology. ” 5th Edition, Spectrum Books Limited, Ibadan, PP 22-23, 2007. [14] D. R. Livingstone. “Oxidative stress inaquaticorganism inrelation to pollution and agriculture” Revue de MedecineVeterinaire, VOL 154, PP 427-430, 2003. [15] J. C. Van Dyk, G. M. Pieterse and J. H. J. Van Vuren. “Histological changes in liver of Oreochromis mossambicus (Cichlidae) after exposure to cadmium and zinc.” Ecotoxocol. Environ. Safety, vol 66, pp 432- 440, 2007. [16] S. Hayat and M. Javed. “Regression studies of planktonic productivity and fishyieldwithreferenceto physico-chemical parameters of the ponds stocked
  • 6. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD29989 | Volume – 4 | Issue – 2 | January-February 2020 Page 284 with sub-lethal metal stressed fish. ” International Journal of Agriculture and Biology, vol 10 pp 561565, 2008. [17] J. C. Akan, M. Salwa B. S. Yikala and Z. M Chellube. “Study on the Distribution of Heavy Metals in Different Tissues of Fishes from River Benue in Vinikilang, Adamawa State, Nigeria. ” British Journal of Applied Science and Technology, VOL. 3(4), PP 1548-1557, 2013. [18] A. O. Igwegbe, A. N. Charles, C. C. Elizabeth and H. B. Mamudu. “Effects of season and location on heavy metal contents of fish species andcorrespondingwater samples from Borno State of Nigeria. ” GlobalAdvanced Research Journal of Medicine and Medical Science, vol. 3(3), pp 064-075, 2014. [19] Khare, S and Singh, S. “Histopathological lessons induced by copper sulphate and lead nitrate in the gill of freshwater fish Nundus.” J . Ecoto xicol. Environ.Mar. Poll. Bull. , vol. 6: pp 5760-5760, 2002 [20] G. Nussey, J. H. J. Van Vuren, and H. H. Du Preez. “BioaccumulationofChromium,Manganese,Nickel and Lead in the tissues of the Moggel, Labeoumbratus (Cyprinidae),fromWitbank Dam,Mpumalanga.”Water South Africa,vol. 26, pp 269-284, 2000. [21] F. A. Oguzie. “Heavy metals in Fish, WaterandEffluents of lower Ikpoba River in Benin City, Nigeria. ” Pakistan Journal of Science and Industrial Research, VOL 46, pp 156-160, 2003.