CHARACTERIZATION AND QUANTIFICATION OF OESTROGENIC ENDOCRINE DISRUPTORS IN LAKE VICTORIA IN TANZANIA, UGANDA AND

KENYA

Mdegela, R.H1., Mbuthia, P.G2., Byarugaba, D.K3., Mabiki, F1., Msigala, S1., Waweru, P. K2., Saphan, M3 and Mwesongo, J1

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Venue: Entebbe, Kampala, UgandaDate: 25th to 28th May 2014VicRes Scientific Conference

LAKE VICTORIA

Sub-optimal productivity of fish in Lake Victoria • Climate challenge –

global warming• Overfishing• Diseases – Infectious

and non-infectious• • Pollution - Point and

non-point sources

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THREATS FROM POLLUTION

Increased human population

Unplanned (informal) settlements

Poor management of wastes

Direct disposal of wastes in water bodies (Rivers & Lake)

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POLLUTANTS OF CONCERN

Conventional group of pollutants and challenges

Pesticides Heavy metals

Emerging group of pollutants and challenges

Endocrine disruptors

Pharmaceuticals

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Justification

Exposure of fish to oestrogenic endocrine disruptors:

Alter normal functions of natural hormones responsible for critical physiological processes

A challenge of great concern worldwide

Many Developed countries, have determined: Sources, concentrations, identities, Health effects in aquatic & terrestrial animals In developing countries, such studies are limited

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Environmentally Important Oestrogens

Oestrogenic hormones include: 17 β -estradiol – β E2 17α-estradiol – α E2Estriol - E3 Estrone - E1 Mestranol - M 17α-ethynylestradiol – EE

Important characteristicsLow water solubilityAssociated with suspended solids

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Aim of the study

To detect and quantify selected oestrogenic endocrine disruptors in water in Lake Victoria in view to assess their contribution to health status of fish

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STUDY AREA: LAKE VICTORIA, MWANZA GULF

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Maganga beach, Nyashishi River, Butimba River, Nyakurunduma River, Mirongo River, Old Igombe Landing site, Butuja, Shede and Kigongo Ferry (9 sites)

Lake Victoria, Napoleon Gulf, Jinja, Uganda

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Rwebitookeisland, Bidico, Wairaka, wanyange, Masese, kakira, wanyange drainage, kirinya-fish industry, kirinya–leatherindustry, NWSC, and area with no human activity (11 site)

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Figure 1: Lake Victoria, Winam Gulf, Kenya

Railway sewage, Tilapia beach, Dunga, Rivers: Kisat, Luanda, Otiako, Nyamasaria, Tako, Wigwa, Auji & Kibos (11 sites)•Homabay landing site, Rivers: Awach, Nyakwanya, Arujo (4 sites)

Collection of water samples

Three water samples were collected from each of the sampling points that included

InshoreIntermediate point 1(00 meters from

inshore)Offshore (500 meters from the inshore)Rivers (100M before entry into lake)At entry point into the lakeAt 500 meters from the river entry points

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Collection of water samples (cont……)

Water samples were collected in clean 2.5 liter glass bottles

Extractions done within 12-24 hours after collection

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Fish samples

Mwanza: Live Tilapia fish were fished where all the water sampling points converged.

In Jinja (Live 44 Tilapia), Kisumu and Homabay (71 Tilapia and 52 Nile

perch) live fish samples were caught from the selected water sampling sites and others purchased from fishermen at the landing sites.

Each fish sampled was examined for gross and histopathological lesions externally and internally.

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Extraction of pollutants in water

Extraction of water samples carried out according to Swart and Pool (2007) with minor modifications

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Detection and quantification

The detection and quantification used Estrogen (El/E2/EE) ELISA KIT

Competitive ELISA technique as per manufacturers recommendations

Total oestrogens (E1, E2 and EE)Estrone(E1), 17-β Estradiol (E2) Ethynylestradiol (EE)

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Results: Validation of the Assay

y = -0.3446x + 1.7173R² = 0.9314

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

0 0.016 0.16 1.6

Abs

orba

nce

(450

nm

)

(a) Concentration of total oestrogens in water extract from a sewage pond effluent (pg/L)

y = -0.2825x + 1.4855R² = 0.9856

0

0.2

0.4

0.6

0.8

1

1.2

1.4

0 0.05 0.15 0.5 3A

bsor

banc

e (4

50 n

m)

(b) Concentration of total oestrogens in standards (µg/L)

Degree of agreement of the two established correlation coefficients was 94.5%

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Results: Total estrogenic EDs (mean ± SD) concentration (pg/L) in water at different sites

50 100 150 200

Inshore

Intermediate

Off shore

Rivers before entry

Rivers at entry

Concentration of total oestrogens (pg/L)

a

b

a

a

a

17

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Results: Total estrogens concentration (ng/L) in Homabay, Kenya

Results: Total oestrogens, Estrone and Estradiol (ng/L) in water from different sites

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Results: Comparison of estrogens levels (μg/L) in Napoleon Gulf and selected waste water sites in Kampala Uganda 20

Summarized Eds Results

Mwanza Gulf:

1. Total oestrogens - 0.39 – 38.9 ng/L

2. Estrone (E1) - 0.37 – 6.36 ng/L

3. 17-β Estradiol (E2) - 0.0021 – 0.12 ng/L

4. Ethynylestradiol (EE) - 0.0026 – 0.032 ng/L

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Summarized Eds Results

Napoleon Gulf1. Total oestrogen -200

- 800 ng/L2. Estrone (E1) - 9.8 -

49 ng/L 3. 17-β Estradiol (E2)-

145 - 305 ng/L 4. Ethynylestradiol

(EE) 45 - 360 ng/L

Kisumu1. Total Estrogen - 0.57 – 64.21

ng/L Homabay –

1. Total Estrogen- 0.35 – 3.63ng/L

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Gross Abnormalities in Fish1.Missing tail/fins - 18.3 %2.Missing eye/opacity - 17.33.Deformed body - 11.54.Deformed head (jaw, face) heart - 8.75.Kinked back and tail - 6.76.Abnormal body color - 3.87.Growth on back - 3.88.Operculum absent - 2.99.Missing scales - 1.910. Dropsy - 1.011.Splenic granulomas/ increased MM12.Cystic formation in gonads - 2.413.Aspermia, cystic and atretic gonads

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Dorsal fin starting from the middle of the back

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Prevalence of lesions in gonads

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Prevalence of lesions in gonads

Cysts embedded in the testicular parenchyma: 2.4%Loosely attached fluid filled cysts: 1.2%

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Histopathological Lesions29

Discussion/Conclusions

All water samples analyzed were positive for total, E1, E2 and EE oestrogenic endocrine disruptors

Some areas had low concentrations, similar finding to other studies (Campbell et al., 2006)

Although low, their potential impact on fish health must be considered

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Discussion/conclusions

In all countries highest estrogens concentration were detected in rivers 100 meters before and at entry point into the lake, fish landing sites and water from sewage treatment plants.

Significant difference (P<0.05) was observed for estrogens levels in river water samples and different lake sampling points.

This study shows existence of oestrogen EDs at different concentrations, and rivers are the main sources of pollutants into Lake Victoria.

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Conclusions/recommendations

Findings from this study therefore calls for intervention that would minimize the discharge of domestic waste water into the rivers and eventually into the lake

Measures for treatment of polluted water before being discharged into the lake should be among target strategic points

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Conclusions/recommendations Oestrogenic pollutants be considered

among emerging pollutants in developing countries

Further studies Profiling of oestrogenic pollutants Effect studies of oestrogenic endocrine

disruptors in fish

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ACKNOWLEDGEMENTs

SIDA – VICRES PROJECT IUCEA UNIVERSITY OF NAIROBI SOKOINE UNIVERSITY AGRICULTURE MAKERERE UNIVERSITY ALL LAKE VICTORIA – FISHERIES

STAKEHOLDERS

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THANK YOU FOR LISTENING & SUPPORT

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