ABSTRACT
The aim of this study is to determine the effects of Lead Nitrate and crude oil on the juveniles of the African catfish Clarias gariepinus. The study was based on results of acute toxicity tests, haematological, histopathological, and enzyme analysis. The acute toxicity tests lasting 96 hours were performed on the Clarias gariepinus of mean weight 138 ± 12g and mean total length of 28 ± 1.5cm. The LD50 of Lead Nitrate was 57.5mg/L while LD50 of crude oil was 823.3µl/L (ppm). The experimental fishes used for the chronic toxicity assay were 27.38 ± 0.43cm – 31.18 ± 3.76cm in mean length and 126 ± 3.22g to 151 ± 0.67g in mean weight. They were divided into groups A, B, C, D, and E of 20 fishes each. Group A was kept as control. Group B was exposed to 20mg/L of Lead Nitrate. Group C was exposed to 35mg/L of Lead Nitrate. Group D was exposed to 300ppm while E was exposed to 600ppm of crude oil. Water quality parameters, Dissolved Oxygen (DO), temperature, and pH were monitored. The body weights of the fishes were measured weekly. The experiment lasted for 70 days. Blood samples were collected for haematological and enzyme analysis before dissection of the gills and liver for histopathological examination. The control group showed significant increase in body weight at (P < 0.05) 150g ± 0.85 to 250g ± 7.07 while the treated groups showed significant reduction in body weight, 151g ± 0.67 to 75g ± 10.61. There was no significant difference (P> 0.05) in the water quality parameters DO, temperature, and pH between the control and treated groups. Significant differences (P<0.05) occurred in haemoglobin, haematocrit, red blood cells (RBC) and white blood cells. No significant differences (P> 0.05) occurred in the RBC indices, mean corpuscular haemoglobin concentration(MCHC), mean corpuscular haemoglobin(MCH), mean corpuscular Volume(MCV). Significant differences (P< 0.05) occurred between the serum enzymes: aspartate transaminase(AST), alanine transaminase(ALT), alkaline phosphatase(ALP) of the control and treated groups. The histological sections of the gills and liver showed histopathological lesions in the treated groups. These include hyperaemia, Oedema, necrosis, degeneration and cellular infiltration. The overall effect of Lead Nitrate and Crude oil on the juveniles of Clarias gariepinus was anaemia and weight loss.
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Lead Toxicity |
15 |
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2.8 |
Crude Oil Toxicity |
17 |
CHAPTER THREE |
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3.0 |
Materials and Methods |
18 |
3.1 |
Determination of LD50 of lead Nitrate and Crude Oil |
19 |
3.2 |
Experimental Design |
22 |
3.3. |
Haematological Examination |
26 |
3.4 |
Enzyme Analysis |
28 |
3.5 |
Histopathological Examination |
28 |
3.6 |
Statistical Analysis |
29 |
CHAPTER FOUR |
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4.0 |
RESULTS |
30 |
4.1 |
Weight of Experimental Fish during Exposed |
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to Lead Nitrate and Crude Oil |
30-33 |
4.2 |
Water Quality Parameters of Exposure Aquaria |
35-37 |
4.3 Haematological Responses of Experimental
Clarias gariepinus after 70 days exposure to Lead
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Nitrate and crude oil |
38-40 |
4.4 |
Enzyme Assay |
41-43 |
4.5 |
Summary of Results |
44 |
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10 |
Histopathological Findings |
46-55 |
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CHAPTER FIVE |
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5.0 |
Discussions |
56 |
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CHAPTER SIX |
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6.0 |
SUMMARY & RECOMMENDATIONS |
64 |
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6.1 |
Summary |
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64 |
6.2 |
Recommendations |
64 |
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References |
66-78 |
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Glossary |
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79-80 |
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Appendices |
81 |
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LIST OF TABLES |
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Table 3.1: |
LD50 of Lead Nitrate |
19 |
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Table 3.2: |
LD50 of Crude Oil |
21 |
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Table 4.1: |
Mean weight of experimental fish |
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during exposure to Lead Nitrate |
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and Crude Oil |
30 |
Table 4.2: |
Water Quality Parameters of Exposure |
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Aquaria |
81 |
Table 4.3 |
Haematological Responses (Mean ± Sem) of |
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Clarias Gariepinus after 70days Exposure to |
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Lead Nitrate and Crude Oil |
82 |
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11 |
Enzyme Assay |
83 |
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LIST OF FIGURES |
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Figures 1: |
Regression lines of changes in mean weight of |
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experimental fish during exposure to Lead |
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Nitrate |
31 |
Figure 2: |
Regression lines of changes in mean weight of |
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experimental fish during exposure to Crude oil |
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32 |
Figure 3: |
Graph showing changes in mean weight of the |
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experimental fish |
33 |
Figure 4: |
Dissolved Oxygen of Exposure Aquaria |
35 |
Figure 5: |
Hydrogen Ion Concentration (pH) of Exposure |
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Aquaria |
36 |
Figure 6: |
Temperature of Exposure Aquaria |
37 |
Figure 7: |
Haemoglobin of Clarias gariepinus after 70 days |
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exposure to Lead Nitrate and Crude oil |
38 |
Figure 8: |
Haematocrit of Clarias gariepinus after 70 days |
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exposure to Lead Nitrate and Crude oil |
39 |
Figure 9: |
Red Blood Cell Count of Clarias gariepinus |
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after 70 days exposure to Lead Nitrate |
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and Crude oil |
40 |
Figure 10: |
SGOT (AST) of Clarias gariepinus after |
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70 days exposure to Lead Nitrate and |
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Crude oil |
41 |
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12 |
SGPT (ALT) of Clarias gariepinus after 70 days |
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exposure to Lead Nitrate and Crude oil |
42 |
Figure 12: |
Alkaline Phosphatase of Clarias gariepinus |
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after 70 days exposure to Lead Nitrate |
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and Crude oil |
43 |
LIST OF PLATES |
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Plate 1: |
Experimental Setup |
24 |
Plate 2: |
Gill Section of Control Fish |
46 |
Plate 3: |
Gill Section of 20mg/l Pb (NO3)2 Treated |
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Group |
47 |
Plate 4: |
Gill Section of 35mg/l Pb (NO3)2 Treated |
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Group |
48 |
Plate 5: |
Gill Section of 300ppm Crude Oil treated |
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Group |
49 |
Plate 6: |
Gill section of 600ppm Crude Oil Treated |
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Group |
50 |
Plate 7: |
Liver section of Control Fish |
51 |
Plate 8: |
Liver section of 20mg/l Pb(NO3)2 group |
52 |
Plate 9: |
Liver section of 35mg/l Pb (NO3)2 group |
53 |
Plate 10: |
Liver section of 300ppm crude oil group |
54 |
Plate 11: |
Liver section of 600ppm crude oil group |
55 |
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13 |
INTRODUCTION
Trace amounts of metals occur naturally in water, however waste water from mining, chemical industries and Agriculture, etc, are the main sources of pollution. Additionally, rain contaminated by burning fossil fuels is another source of pollution (Pitter, 1999). In fish, the route of heavy metal entry is either through the gills or mouth. The blood carries these heavy metals to different organs or systems hence haemotological studies and histopathological examinations have been considered as valuable diagnostic tools in clinical biochemistry, physiology, genetics and medicine. For many years, these have been used to investigate disease or metabolic alterations (Bansal et al., 1979).
Physico-morphological changes in the blood indicate the changes in the quality of the environment, therefore blood parameters are important in diagnosing the functional status of animals exposed to toxicants (Borane and Zambare, 2006).
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Fishes are particularly sensitive to water borne environmental contamination and are recognized as a useful model for indicating water quality (Mathis and Kervern, 1975).
Lead, a biologically non-essential metal is relatively abundant in nature and of extensive use in modern times (Todd et al.,
1996). The build up and transportation of lead in water, atmosphere and sediment result in bioaccumulation of the metal in various pockets of food chain. Sub lethal concentrations of lead cause toxicity which results into oxidative damage in fish tissues. It produces oxidative stress through the generation of free radicals in which reactive oxygen species (ROS) are most important in causing damage to cells and tissues (Verma and Belsare, 2005).
Crude oil pollution is common all over the world, particularly endemic in countries whose economies are dependent on the oil industry. In Nigeria oil industry, operations are both onshore and offshore and the oil terminals as well as most refineries are located in the Niger Delta region, hence more than 90 percent of oil related activities take place in this region (Imevbore and Adeyemi, 1981). According to the River State Environmental
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