.

ACTIV E BIOMONITORING OF TRACE HEAVY METALS USING FISH (Oreochromis niloticus) AS BIOINDICATOR SPECIES:THE CASE OF NAKIVUBO WETLAND ALONG LAKE VICTORIA. Z. Birungi1, B.Masola2, M.F. Zaranyika3, I. Naigaga4, B. Marshall5 Department of Civil Engineering, University of Zimbabwe, P.O. MP 167, Mount Pleasant, Harare, Zimbabwe 2 Department of Biochemistry, University of Zimbabwe, P.O. MP 167, Mount Pleasant, Harare, Zimbabwe 3 Department of Chemistry, University of Zimbabwe, P.O. MP 167, Mount Pleasant, Harare, Zimbabwe 4 Warm department, Veterinary Sciences, Makerere University, P.O Box 7062, Kampala, Uganda 5 Department of Biological Sciences, University of Zimbabwe, P.O. MP 167, Mount Pleasant, Harare, Zimbabwe ABSTRACT The Nakivubo wetland, which feeds Lake Victoria at Murchison bay, has become severely degraded over recent years and is particularly threatened by the expansion of industry, settlement and cultivation on its fertile fringes. The aim of this study was to actively biomonitor selected trace heavy metals using Nile tilapia (Oreochromis niloticus). Nile tilapia was set in cages in Murchison bay for six weeks, and sampling was done every 2 weeks for active biomonitoring and weekly for physico-chemical variables. Fish tissue was dissected and gills, liver and muscle removed for heavy metal analysis. Water samples from each site were analysed using AAS to determine metal concentrations of Zinc, Copper, Chromium and Manganese. Site 4 had the highest average NH4-N of 14.2812.73 mg/l which was above effluent standards for Uganda. From the findings, there were significantly higher concentrations of heavy metals in fish than in water (p>0.01). Fish kills were recorded highest for sites 4 (55%), 5 (77.5%) and 6 (60%) in less than six weeks indicating high levels of pollution. Copper had the highest bioaccumulation in tissue when compared to Zn, Mn and Cr, and the order of bioaccumulation was; gills>muscle>liver. Gills had highest copper accumulation with an average concentration of 11.70.42g/mg after 6 weeks. The Nakivubo wetland is not effective in cleaning up the pollution loads from the city of Kampala which could lead to risks of human health and eutrophication of Lake Victoria. There is therefore a need to upgrade the wetland through resettling people in parts of the degraded wetland and regular monitoring of polluters. There is also need to incorporate postmortem studies and biomarker analysis since the later can give early warning of pollution before fish kills are observed.
1

Key words: accumulation, fish, heavy metals, wetland.

1. Introduction Heavy metals normally occurring in nature are not harmful to the 1

environment, because they play an essential role in tissue metabolism and

growth

of

plants

and

animals

entire ecosystem. This study was aimed at determining in heavy Nile metal tilapia

(Amundsen et al., 1997). But severe metal imbalances are toxic and marginal imbalances contribute to deformities and impede health. Metals of major interest in bioavailability studies, as listed by the U.S. Environmental Protection Agency are Al, As, Be, Cd, Cr, Cu, Hg, Ni, Pb, Se, and Sb (McKinney and Rogers, 1992). Other metals that are presently of lesser interest to the EPA are Ag, Ba, Co, Mn, Mo, Na, Tl, V, Fe and Zn. These metals were selected because of their potential for human exposure and increased health risk. A study of the effects of effluents on the aquatic ecosystems has a high ecological

accumulation

(Oreochromis niloticus) using active biomonitoring (ABM) which is a

potentially useful method for monitoring toxicity of effluents. 2. Materials and Methods 2.1. Study area The study was carried out in Nakivubo wetland at inner Murchison bay on Lake Victoria, Uganda (map. 1 and 2). It is the largest of the twelve main wetland areas in Kampala covering approximately 5.29 km2, and has a total catchment extending over 40 km2 (COWI/VKI 1998). The Nakivubo River and its tributaries provide the main drainage channel for Kampala, carrying wastewater from the city centre, from the industrial area and from residential areas to the north-west of the city. Although Inner Murchison Bay is fed by 8 sub-catchments, by far the largest contribution to its nutrient load is made from the Nakivubo catchment. Six sites (1, 2, 3, 4, 5, and 6) were chosen for active biomonitoring and physico-chemical investigation of water pollution (Map 2).

relevance because effluents constitute a direct and continuous input of pollutants in these ecosystems (Smolders et al., 2003). The heavy metal concentration in tissues reflects past exposure via water and/or food and it can demonstrate the current situation of the animals before toxicity affects the ecological balance of populations in the aquatic environment (Forstner and Wittmann, 1983). Since water is the basis for all organisms and ecosystems, protection of aquatic

resources is essential in protecting the

Map 1: The location of Lake Victoria in Kampala, Uganda

2.2 Sampling Protocol Juvenile fish (Oreochromis niloticus), 60100 g were obtained from the fish farm of Aquaculture Research and Development Centre (ARDC, Uganda) and put in hapas (cages) at selected sampling sites (Fig 1). The sample size was 40 fish for each cage with the reference site at the fish farm. Sampling was done weekly for physicochemical parameters and every two weeks for active biomonitoring in a period of six weeks. During sampling, four fish and water samples were taken from each site and

transported to the biochemistry laboratory at Makerere University. Each fish was

weighed, total and standard length measured before dissection. The fish that did not survive field exposure were thrown away. Dissection was carried out on an aluminum foil work place using stainless steel tools and wearing surgical gloves. Muscle, gills and liver were removed for metal analysis. After dissection, the tissue samples were wrapped in aluminum foil and frozen at -80
0

prior

to

metal

analysis.

The

concentrations of metals which include

copper, chromium, zinc, manganese were determined in the water samples using Perkin-Elmer 2380 Atomic Absorption

including the control using the instruments indicated temperature in parentheses: (WTW pH pH 330 and meter),

Spectrophotometer (AAS).

dissolved oxygen (WTW Oxi 330 meter), total dissolved solids and conductivity (WTW LF 330 meter).Other chemical variables were measured with a Hach water analysis kit (DR/2010 portable data logging spectrophotometer) using filtered water samples (Whatman GF 47mm filters). The methods are summarized in the

manufacturers manual and described in detail in (Bartram and Ballance

1996).Turbidity, nitrate nitrite ammonianitrogen calcium, potassium, sulphates and chlorine were determined. 2.3 Preparation of fish tissue for analysis The Food and Agricultural Organization (FAO) method (1983) involving the

digestion of the sample in an open beaker on a hot plate was used. . Prior to use, all glass ware was previously soaked in dilute nitric acid for 24 hours and then rinsed with deionised water. Fresh weight of gills, muscle and pooled liver samples from each site was weighed and placed in beakers. Fig. 1: One of the experimental cages set in Murchison bay along Nakivubo wetland Gills filaments were removed from the cartilaginous gill arches before being

weighed. Using the FAO method which The following physico-chemical variables were determined in situ at all the seven sites involves 10g fresh weight in 15ml of freshly prepared 1:1 (v/v) nitric acid- hydrogen 4

peroxide, the varied weight of the pooled samples upto 2 g were calculated backwards to coincide with this method. The beakers were covered with a watch glass and set aside for about 15minutes in order to allow the initial reaction to subside. The beaker and its contents were heated at a temperature of the hot plate not exceeding 1600C.Boiling was done for 30min and subsequently reducing the volume. The contents of the beaker were transferred into a 25ml

The stock solutions were used to calculate the required volumes for concentrations of 2,4,6,8 and 10 ppm in final volumes of 100 ml. The formular used was; C1V1 = C2V2 where,

C1 is standard concentration of 1000 ppm V1 is the required volume to be calculated. C2 are different concentrations of 2,4,6,8 and 10 ppm V2 is the volumetric flask used (100ml) After calculating the required volumes for each metal (0.2, 0.4, 0.6, 0.8 and 1ml) and pippeting these ampunts into 100ml flasks,

volumetric flask and diluted to the mark with deionised water. 2.4 Preparation of stock standard solutions and analysis Stock standard solutions were prepared for all the four metals. For 1000 mg/l of chromium, 3.735g of potassium chromate was dissolved in deionised water and diluted to 1 liter with deionised water. For 1000 mg/l of copper, 1g of copper metal was dissolved in a minimum volume (1:1) nitric acid and then diluted to 1 liter with 1% (v/v) nitric acid. For 1000mg/l of manganese, 1g of Mn metal was dissolved in a minimum volume of (1:1) nitric acid and diluted to 1 liter with 1% (v/v) hydrochloric acid (HCl). For 1000 mg/l of Zinc, 1g of Zn metal was dissolved in a minimum volume of (1:1) HCl and diluted to 1 liter with 1% (v/v) HCl.

the flasks were filled to the mark with deionised water. These calculated volumes were then used to set up a standard linear graph which was used to determine the concentration of metal in samples of gill, liver and muscle from each site. A Shimadzu AA 6401F Atomic Absorption

Spectrophotometer (ASS) was used to determine the levels of copper, manganese, zinc and chromium in these tissues. Average concentrations were then calculated and converted from mg/l to g/mg. 1ppm = 1mg/l = 1000 g/l,

For fish, g (mass of metal)/g (mass of fish sample) was determined and then converted to g/mg. 2.5 Statistical Analysis The physico-chemical variables for water were analysed using Microsoft excel to determine mean and standard deviation and graphs were plotted with the aid of 2004 SYSTAT software (Sigma plot (9)). To do a univariate analysis of only two groups (mean concentrations of trace heavy metals in water and fish), the t-test was employed. Analysis of Variance (ANOVA) was used to analyse bioaccumulation of heavy metals in tissue. 3. Results 3.1 Physical- Chemical Variables of Water Table 1 shows physico-chemical conditions at the six sites in Murchison bay along Nakivubo wetland, Kampala. Site 7 was the control on a fish farm at Aquaculture Research and development Center from 10thJanuary-21stFebruary, 2006. All data were presented as means standard deviation (S.D). E.cond = Electrical

(mg/l), NO2- = Nitrite (mg/l), Cl- = Chloride (mg/l), SO4- = Sulphate (mg/l), K+ = potassium (mg/l), Ca2+ = Calcium (mg/l). The pH range and temperature for all the seven sites was within permissible effluent standards for Uganda 6-8 and 20-350C respectively. Site 3, 4 and 5 had the lowest average dissolved oxygen levels. Site 4 had the highest average NH4-N of 14.28 12.73 mg/l.

3.2 Nile Tilapia as Bioindicators of Water Pollution Forty Nile tilapia were stocked in cages in different sites at Murchison bay which was highly polluted from sewage and industrial effluent. Site 5 had the highest number of fish kills (77.5%), followed by site 6 (60%) and site 4 (55%).100% of the fish stocked in site 7 survived.

3.3 Comparison of Heavy Metals in Water and Nile Tilapia A preliminary test was carried out to test the hypothesis of heavy metal accumulation in fish is significantly higher than heavy metal determination in water for copper, using T-

conductivity (s/cm), Temp = Temperature (0C), DO = Dissolved Oxygen (mg/l), TDS= Total dissolved solids (mg/l), N-NH4= Ammonia-nitrogen (mg/l), NO3- = Nitrate

manganese, zinc and chromium

test. Results from analysis showed that the

mean amount of all the tested metals in fish was greater than that in water (p<0.001).

3.4 Comparison of heavy metals in fish tissue, sites and period After carrying out ANOVA, the results for copper showed significant differences in tissue and period (p< 0.05) as shown in table 3. Site 1 showed the highest accumulation of copper after a period of 2 weeks in liver (20 g/mg) but which reduced to 2 g/mg after 6 weeks

Table1: Physico-chemical variables at the seven sites in Murchison bay and the fish farm, n =5
Variables E.cond pH Temp DO TDS Turbidity N-NH4 NO2NO3 Cl
-

site 1 12917.92 6.50.33 25.721.61 2.862.36 80.4817.64 21.6013.45 3.181.32 0.030.05 0.050.04 1.130.30 4.220.77 4.620.83 8.542.67

site 2 12616.86 7.30.53 26.401.38 2.782.12 78.4816.77 42.6025.99 3.061.75 0.10.20 0.991.33 1.190.46 4.380.88 4.580.62 8.501.76

site 3 16363.02 6.90.48 25.71.41 1.801.02 104.2542.19 22.6021.36 8.075.30 0.220.30 1.351.96 0.970.23 3.500.75 6.263.49 9.843.03

site 4 18999.23 6.90.17 25.61.34 1.550.54 122.7564.5 84.75136.42 14.2812.73 0.150.27 0.170.13 1.080.27 3.531.17 7.005.07 12.605.08

site 5 21399.17 6.70.08 25.82.01 1.230.06 129.348.88 201.67237.39 8.495.65 0.120.17 1.641.40 1.240.33 4.670.50 8.034.68 13.072.83

site 6 22275.97 6.80.34 26.731.62 3.600.72 144.3049.38 40.6734.20 7.403.79 0.150.12 1.180.96 0.990.38 3.930.51 4.634.85 12.604.24

site 7 (control) 825.45 7.91.16 24.741.55 2.361.82 50.827.65 235.5049.13 5.033.40 0.050.07 0.040.06 11.810.00 3.320.217 3.580.421 6.300.70

SO4 K+

Ca2+

Table 2: Survival and death rate of Nile tilapia set in cages at the seven sites in Murchison bay and fish farm from 10th January-21st February 2006.
Fish kills/ week sites 1 2 3 4 5 6 7 2 0 2 2 5 15 20 0 3 1 2 4 2 16 2 0 2 0 0 0 4 1 1 1 7 5 1 2 1 8 6 1 1 1 Av. fish kills 4 9 9 22 31 24 0 Survival/40fish 36 31 31 18 9 16 40 % fish kills 10 22.5 22.5 55 77.5 60 0 %Survival 90 77.5 77.5 45 22.5 40 100

(Fig. 2). There was bioaccumulation of copper in muscle for all the sites except 4, 5 and 6 which had no fish left by the sixth week (Fig. 3). The results for manganese showed significant differences in tissue, site and period (p< 0.05) as shown in Table 4. Site 2 had the highest bioaccumulation of Mn in liver with an average concentration of 4g/mg after a period of 6 weeks (Fig.5). Sites 3 showed the highest bioaccumulation after a period of 6 weeks in gills with an average concentration of 2 g/mg (Fig.7).

After carrying out ANOVA, the results for zinc showed significant differences in tissue p< 0.05 (Table 5). Site 3 had the highest bioaccumulation in gills after a period of 6 weeks with an average concentration of 10g/mg (Fig.10). Site 6 also showed bioaccumulation in muscle after a period of 4 weeks with an average of 6g/mg (Fig. 9).

The results

for

chromium

showed

significant differences in tissue and period, p< 0.05 (Table 14). These results indicated bioaccumulation although on average chromium recorded the lowest levels in Oreochromis niloticus as compared to other heavy metals (Table 7, 8, 9).

Comparison of heavy metals basing on tissue type and period only, showed that copper had the highest bioaccumulation as compared to other heavy metals in all tissue (Table 7, 8, 9).

Table 3: Results of copper between tissue, site and period from ANOVA

Source Model Tissue Site Period Error Total

Type 111 sum 6.905 2.965 3.637 0.521 11.62 18.526

a

df 11 2 6 2 40 51

Mean Square 0.628 1.482 0.606 0.26 0.291

Freq f 2.161 5.103 2.087 0.896

Significant Difference 0.038 0.011 0.076 0.416

Source Model Tissue Site Period Error Total

Type 111 sum 5.157 1.326 1.163 2.576 5.895 11.051

a

df 11 2 6 2 40 51

Mean Square 0.469 0.663 0.194 1.288 1.47

Freq f 3.181 4.499 1.315 8.741

Significant Difference 0.004 0.017 0.273 0.001

Table 4: Results of manganeser between tissue, site and period from ANOVA

Table 5: Results of zinc between tissue, site and period from ANOVA

Source Model Tissue Site Period Error Total

Type 111 sum 15.572 2.371 2.589 1.947 5.662 21.233

a

df 11 2 6 2 39 50

Mean Square 1.416 1.186 0.432 0.973 0.145

Freq f 9.752 8.167 2.973 6.706

Significant Difference 0 0.001 0.017 0.003

10

Table 6: Results of chromium between tissue, site and period from ANOVA

Source Model Tissue Site Period Error Total

Type 111 sum 12.253 8.517 0.456 2.436 3.037 30.056

a

df 10 2 6 2 40 51

Mean Square 1.225 4.258 0.076 1.218 0.076

Freq f 16.137 56.085 1.002 16.043

Significant Difference 0 0 0.438 0

11

25

4
20

accumulation

accumulation

15

10

1
5

0
0 0 1 2 3 4 5 6 7 8

s ite s

s it e s

Fig. 2: Copper accumulation (g/mg) in liver for a period of 2, 4 and 6 weeks

12

Fig. 5: Manganese accumulation (g/mg) in liver for a period of 2, 4 and 6 weeks

10

1 .8 1 .6

accumulation

1 .4 1 .2

accumulation
0 1 2 3 4 5 6 7 8

1 .0 0 .8 0 .6

2
0 .4 0 .2 0 .0 0 1 2 3 4 5 6 7 8

sites

s ite s

Fig. 3: Copper accumulation (g/mg) in muscle for a period of 2, 4 and 6 weeks

Fig. 6: Manganese accumulation (g/mg) in muscle for a period of 2, 4 and 6 weeks

18 16

2 .5

2 .0

14
accumulation

aaccumulation

12 10 8 6 4 2 0 0 1 2 3 4 5 6 7 8

1 .5

1 .0

0 .5

0 .0 0 1 2 3 4 5 6 7 8

s ite s

sites
2weeks 4weeks 6weeks

Fig. 4: Copper accumulation (g/mg) in gills for a period of 2, 4 and 6 weeks

Fig. 7: Manganese accumulation (g/mg) in gills for a period of 2, 4 and 6 weeks

12

12
0 .8

10
0 .6

accumulation

accumulation

0 .4

4
0 .2

0 .0

0 0 1 2 3 4 5 6 7 8

s it e s

s it e s

Fig. 8: Zinc accumulation (g/mg) in liver for a period of 2, 4 and 6 weeks

7

Fig. 11: Chromium accumulation (g/mg) in liver for a period of 2, 4 and 6 weeks

1 .2

1 .0

accumulation

0 .8

accumulation

0 .6

0 .4

0 .2

0 0 1 2 3 4 5 6 7 8

0 .0 0 1 2 3 4 5 6 7 8

s ite s

s it e s

Fig. 9: Zinc accumulation (g/mg) in muscle for a period of 2, 4 and 6 weeks

Fig. 12: Chromium accumulation (g/mg) in muscle for a period of 2, 4 and 6 weeks

12

1.2

10

1.0
8

accumulation

0.8
6

accumulation
0 1 2 3 4 5 6 7 8

0.6

0.4
2

0.2

s ite s
2w eeks 4w eeks 6w eeks

0.0 0 1 2 3 4 5 6 7 8

site s

Fig. 10: Zinc accumulation (g/mg) in gills for a period of 2, 4 and 6 weeks

Fig. 13: Chromium accumulation (g/mg) in gills for a period of 2, 4 and 6 weeks

13

Table 7: Accumulation of different trace heavy metals in liver (g/mg), n=7

period 2 weeks Heavy metal Copper Manganese zinc chromium Copper Manganese zinc chromium Copper Manganese zinc chromium mean 2.3 0.23 0.58 0.27 4.88 1.19 0.16 0.16 1.06 0.41 0.4 0.1 0.89 0.2 0.230.48 0.87 0.34 0.32 0.14 0.8 0.4 0.36 0.13 SD 0.6 0.71 3.16 0.1 1.07 0.27 0.53 0.13 0.91 0.38 1.05 0.35 Min 1.43 0.16 0.2 0.05 0 0 0 0 0 0 0 0 Max 3.33 1.8 9.77 0.35 3.3 0.72 1.46 0.44 2 1.06 2.84 0.74

Table 9: Accumulation of different trace heavy metals in gills (g/mg), n=7

period 2 weeks Heavy metal Copper Manganese zinc chromium Copper Manganese zinc chromium Copper Manganese zinc chromium mean S.E 7.71 3.75 0.34 0.06 0.53 0.11 0.17 0.05 3.19 0.76 0.49 0.17 0.41 0.87 0.41 0.07 11.7 0.42 0.35 0.25 1.69 1.46 0.5 0.18 SD 9.93 0.15 0.29 0.14 1.97 0.44 0.23 0.19 1.12 0.66 3.87 0.48 Min 3 0.03 0.03 0.03 0 0 0 0 0 0 0 0 Max 30 0.52 0.86 0.42 5 1.33 0.69 0.58 2.5 1.82 10 1

4 weeks

4 weeks

6 weeks

6 weeks

Table 8: Accumulation of different trace heavy metals in muscle (g/mg), n=7

period 2 weeks Heavy metal Copper Manganese zinc chromium Copper Manganese zinc chromium Copper Manganese zinc chromium mean S.E 10 0 3.3 0.05 1.09 0.26 0.11 0.01 1.1 0.1 0.69 0.19 1.99 0.77 0.44 0.1 0.57 0.2 0.16 0.07 0.58 0.27 0.56 0.2 SD 0 0.14 0.69 0.04 0.25 0.5 2.02 0.25 0.54 0.19 0.71 0.53 Min 1 0.18 0.05 0.05 1 0 0 0 0 0 0 0 Max 1 0.56 1.98 0.16 1.67 1.65 6 0.79 0.01 0.45 1.62 1.12

4 weeks

6 weeks

14

4. Discussion The low levels of dissolved oxygen in sites 3, 4 and 5 could have been due to the high organic load from the wetland which requires oxygen for decomposition of algae. When decomposition rates are high, then the concentration of dissolved oxygen can decrease to the point of affecting the ability of other oxygen-dependent organisms to survive. Site 6 had the highest DO levels and this could have been attributed to the fact that it was measured last during sampling when the temperature and light were high. Site 7 had NH4-N levels higher than sites 1 and 2 which could be due to decay of grass. The fish pond had dykes with planted grass which was occasionally slashed and in the process some of it would end up in the pond. Occasionally water hyacinth was found in the bay. This water weed is an indication of eutrophication due to the rich organic nutrients coming from the wetland. Losses of fish in Sites 5 and 6 could have been due to clogging of the cages and thus not allowing entry of water which helps improve dissolved oxygen levels and

provision of food. In some instances, the cage in sites 5 was shifted by wind to shallow water which could have lead to suffocation of fish. In addition cages in sites 3, 4, 5 and 6 were occasionally found surrounded by the water hyacinth reducing circulation of water. This weed could have been blown by the wind towards the cages. In aquatic ecosystems fish are regarded as good representative indicators of overall system health due to their relatively high position in the food chain (Adam et al, 1993).Tilapias are hardy fish for culture and can be grown in very harsh environments. This was confirmed with Oreochromis niloticus which were able to survive in sites 1, 2 and 3 up to six weeks despite the extremely high pollution loads in Murchison bay. However, for successful management it is necessary to know their tolerance limits. At least occasional checks should be made of water temperature, salinity, dissolved oxygen and acidity (pH), to ensure that values remain in the range known to be good for tilapia growth. Tilapia does not thrive in low water temperatures but are very tolerant

Eutrophication leads to episodes of oxygen deficiency. In addition, a lot of dead algae were observed on the surface of water which becomes poisonous leading to fish kills. During the fourth and sixth week of sampling, there was bad odour and colour in site 5 and 6 implying discharge of raw sewage into the bay.

15

to high temperatures. Activity and feeding of tilapias are reduced below 20C. The dissolved oxygen (DO) limit for Nile tilapia has been found to be 0.01 mg/l (Nadla and Pickering, 2004). Although tilapia can survive very low oxygen conditions, it does not mean they will grow well. This was observed after four weeks in sites 4, 5 and 6 as the fish were stunted and had flat stomachs indicating little feeding. The DO level for good growth of fish should ideally be above 2mg/l (Nadla and Pickering, 2004). Tilapia can survive in wide range of water acidity- pH between 3.5 and 12. However, the recommended pH for good growth of tilapia is 6.5 to 9. Acidic water (with a low pH) will not support the growth of phytoplankton, zooplankton and detritusdigesting bacteria. All these organisms are important as they collectively provide an environment conducive for fish growth. The pH for all the sites was within acceptable range for growth of tilapia.

dissolved oxygen below required levels for survival of cichlids (2-3 mg/l), and site 6 had the highest electrical conductivity. The bioaccumulations observed in site 7 could be due to the upcoming developments around the fish farm but in general the external environment was acceptable as compared to the sites in the bay.

In general, bioaccumulation of copper in tissue was recorded highest in the order gills>muscle>liver after a period of 6 weeks. Gills are central in the uptake of dissolved substances from the water being the prime target for the toxic action of waterborne metals such as Cd, Cu and Zn (Soto et al, 2003).This fact may explain the highest accumulation in gills. The high levels of copper in the liver can be ascribed to the binding of copper to metallothioneins (MT), which serves as a detoxification mechanisms (Hogstrand and Haux, 1991). Lloyd (1992) found that the main environmental factor which affects copper toxicity in water is

The accumulation of copper reduced after a period of six weeks which could imply that the liver is able to eliminate copper. This could be attributed to the external

calcium concentration. This is as a result of the competition that occurs between these two ions for binding sites in the tissue of the gills and other organs. This could explain the low levels of copper in gills for sites 4, 5 and 6 which had high calcium levels (fig. 6 and 9). Copper in the alkaline conditions

environment, for example site 4 had the highest NH4-N which predisposes fish to bacteria gill disease, sites 4 and 5 had low

16

precipitates as the carbonate and is non toxic (Dallas and Day, 1993).As the majority of the surface water pH values recorded in this study were above 6, most of the copper present may well have been rendered biologically unavailable. Manganese is an important component of many metabolic processes, including the functioning synthesis cholesterol of of and flavoproteins and the

In general, bioaccumulation of zinc in tissue was highest for gills>liver>muscle.

According to Moor and Ramamurti (1987), zinc induced negative changes when

penetrating fish gills. Gills are the main route for metal penetration from the water. The results supported their opinion that the main route of zinc entry into the fish was via water. During the whole period of the study, the lowest zinc loads were found in the muscles. In their studies, Kroupa and Hartvich (1990) also reported low zinc levels in muscles.

mucopolysaccharides, haemoglobin (DWAF,

1996). The toxicity of manganese is not dependent on the total manganese

concentration, but the concentration of oxidized manganese which is available. The presence of high concentrations of chlorides, nitrates and sulphates solubility, may increase both When zinc in the water rises to a level where the amount entering the organism through the gills exceeds the requirement for this metal, the surplus has to be excreted (Lloyd, 1992).This could explain the lowest levels of zinc in gills from results. High levels of zinc leads to further gill damage involving the separation of epithelium, enlargement of central and marginal channels, occlusion of central blood spaces, and results in

manganese

increasing

aqueous mobility and uptake by plants (Clement et al., 1985). Manganese

accumulated in plant material will also provide a source for dissolution during decomposition. Higher concentrations of Ca2+ ions will compete with Mn ions for absorption through the gill surface thus decreasing the bioavailability of manganese to fish (Seymore et al, 1995). This could explain the low levels of manganese recorded in gills for sites 4 and 5 which had high calcium levels (Table 1 and Fig. 7).

decreased oxygen consumption, the ability to transport ions across the gill surface and an increase in hypoxia ventilation frequency (Lloyd, 1992).

Zinc

can

bioaccumulate

in

the

liver

reflecting the multi-functional role of the

17

liver

in

the

detoxification binding) and

(through storage

chromium and this could be due to the upcoming developments like planting of flowers on commercial scale. Acknowledgements: I would like to

metallothioneins

processes (Carpene et al, 1990). It has the ability to bind to metallothioneins, although Cu has a greater affinity for proteins and is able to displace zinc (Roch et al, 1985). This could explain the low content of zinc in liver as compared to copper in the results. slightly basic, environments, In

acknowledge water net which sponsored this research. References Adam, S.M., Brown, A.M. and Goede, R.W (1993). A quantitative health assessment index for rapid evaluation of fish condition in the field. Transactions of the American Fisheries Society. 122: 63-73. Amundsen, P., Staldvik, F. J., Lukin, A. A., Kashulin, N. A., Popova, O. A. and Reshetnikov, Y.S (1997). Heavy metal contamination in freshwater fish from the border region between Norway and Russia. Science of the Total Environment, 201, 371378. Bartram, J and Ballace, R (eds) (1996). Water quality monitoring. E and FN Spon, London, UK. Buhler, D.R., Stokes, R.M and Caldwell, R.S (1977). Tissue accumulation and enzymatic effects of hexavalent chromium in rain bow trout (Salmo gairdneri). J Fish Res Bd Can Carpene, E., Cattani, E., Serrazanetti, P., Fedrizzi, G., and Coretesi, P. (1990).Zinc and copper in fish from natural waters and rearing ponds in Northern Italy. J Fish Biol. 37: 293-299. COWI/VKI, (1998). Kampala Water Quality Monitoring Programme: Murchison Bay Water Quality Project, Report prepared for Ministry of Natural Resources National Water and Sewerage Corporation by COWI in association with VKI, Kampala, Uganda.

anoxic marsh sediment zinc is effectively

immobilized and not bioavailable (Gambrell et al, 1991). Very high abundances of soluble zinc are present under well oxidized conditions and at pH 5 to 6.5, whereas low abundances of soluble zinc are present at pH 8 under all redox conditions and at pH 5 to 6.5 under moderately and strongly reducing conditions (Gambrell et al, 1991). Accumulation of chromium in fish is not a serious problem since the fish are able to eliminate it from the body (Buhler et al, 1977). Chromium readily penetrates gill membranes by passive diffusion and

concentrates at high levels in various organs and tissues. The site of toxic action during lethal chromium exposure to fish varies with different pH values (Van der Putte et al, 1981). Bioaccumulation was highest in sites 1, 2, 3 and 7 but there were no significant differences between these sites (Figs.18, 19 and 20). Site 7 showed bioaccumulation of 18

Dallas, H.F. and Day, J.A (1993). The effect of water quality variables on riverine ecosystems: A review. Fresh water Research Unit, University of Cape Town, Rondebosch, South Africa. DWAF. (1996). Procedures to access effluent discharge impacts. WRC Report No.TT 64-351pp. Clement Associates (1985). Chemical, physical and biological properties of compounds present at hazardous waste sites. Prepared for US Environmental Protection Agency, Washington, DC.312pp. Food and Agriculture Organization of the United Nations, FAO/SIDA (1983). Manual of methods in aquatic environment research. Part 9. Analysis of metals and organochlorine in fish. FAO fish Technical paper 212. (FAO, Rome) Section 2, pp 1420. Forstner, U and Wittmann, G.T.W. (1983). Metal pollution in the aquatic environment. Springer-Verlag, Berlin, pp. 30-61. Gambrell, R.P., Wiesepape, J.B., Patrick, W.H., and Duff, M.C., (1991) .The effect of pH, redox, and salinity on metal release from a contaminated sediment: Water, Air, and Soil Pollution, 57-58: 359-367. Hach Company Procedures Manual (19961999) .Hach Co, Loveland, CO, USA. Hogstrand, C and Haux, C. (1991). Minireview. Binding and detoxification of heavy metals in lower invertebrates with references to metallothinein. Comp. Biochem.Physiol. 100c:383-390.

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