The effects of 96 h sub-lethal concentrations of African locust bean effluent on Oreochromis niloticus were studied using plastic buckets. Packed cell volume (PCY) and histological study of gills and liver were the tissue chemistry parameters investigated. The control buckets contained no effluent. Results revealed increase in ESR (Erythrocyte Sedimentation Rate) decrease in haemoglobin, red blood cell and PCY Yalue of the fish exposed to all sub-lethal concentrations. Damage to liver and gills were noticed in some of the sub-lethal concentrations of all the effluent. Normal situation was observed in all the examined tissues of fish from the control buckets. The African locust bean effluent is toxic on O. niloticus and the lethal concentrations of African locust bean effluent deducted from the 96 hours acute toxicity effect ranged from 3ml

Adebayo, O.T.

Ekpo-Jide, O.

Fagbenro, O.A.

Aquatic Commons

178
MATERIALS AND METHODS
Altogether, 240 live 0. niloticus fingerlings were purchased from Agricultural Development Project
(ADP) fish farm at Alagbaka, Akure. African locust bean effluent was obtained from a local producer
at Oke-Aro Titun m Alcure. The fingerlings were transported to Fisheries laboratory of FUTA and
placed in 100ml of water inside conico-cylindrical plastic containers for 48 hours to acchrnate to
lahoratory conditions. Feeding was discontinued during the acclimation period in order to avoid
pollution of the water with feed wastes. The fingerlings were weighed individually using a top-
loading Mettler balance and randomly distributed into each container containing 100mi of water at ten
fingerlings per container. A range-finding test was conducted using ten containers, each filled with
IOOmlof water prior to the introduction of the effluent. Each of the five varying concentrations (5ml,
lOml, 151111, 20ml and 25ml) of' African locust bean effluent was introduced with a pipette mto the
containers at 09.00 hr and the mixture was stirred. Each stock sulution treatment was replicated twice.
I:'\ITRODUCTTON
Effluent discharges constitute the greatest pollution problem that Nigeria has ever experienced (Sikoki
& Kolo, 1993). The effluent from these agriculture and industrial products, hax conunucd to affect
both mall and aquatic ecosystems directly or indirectly. Industrial effluent discharges exert great
Impact on the aquatic system even at low concentrations. Krous et a/.(1982) and Ofojekwu et
al.( 1990) used different test organisms, different periods of exposure to pollutants, waters of different
quality and different ways of reporting the results. The acute toxicity of Roger" was determined for
the juveniles of the freshwater fish, Nile tilapia (Oreochromis niloticusy by Annunc & Ajike (1999)
and it was suggested that osmoregulatory and respiratory incapacitation through mucus accumulation
and epithelial detachment were the major causes of fish mortality following exposure to Roger".
Exposure of the 0. niloticus to sub-lethal concentrations of malachite green led to severe
physiological impairment of fish after a 10 week exposure period (Omoregie et al., 1992).
The African locust bean (Parkia biglobosa) is a source of protein in diets of Nigerians, and in
the fermented form, it is one of the most important food condiments in the entire savanna region of
West and Central Africa. Locust beans are boiled for about an hour to soften the seed coat for
removal; the hulls are removed followed by washing besides rivers, and boiling for additional 10-12
hours to soften the cotyledons. I'he cotyledons are allowed to undergo wild fermentation for about2
tu 3 days during which the characteristic colour (brown) and odour (amrneruated) are developed. The
effluents generated during African locust bean prcduction may have harmful effect on water bodies as
well as organisms that inhabit such water bodies. The effluent may be introduced deliberately or
accidentally into the aquatic ecosystem during washing besides rivers impairing the quality of the
water and making It unfavourable lor aquatic life. The environment of the fish is often altered by tOXIC
substances and this results in fish kills) especially when the concentration is higher than what the
homeostasis of the fish can control. Information on the effects of sub-lethal and lethal exposure of
freshwater species to African locust bean and its effluent is limited (Ahmed et 01., 2005), hence thrs
study IS to evaluate the effects of sub-lethal concentrations of P. biglobosa effluent on the behaviour.
survival, some haematological parameters. and histology of the gills and liver of 0. niloticus
fingerlings.
ABSTRACT <r
The effects of 96 h sub-lethal concentrations of African locust bean effluent on Oreochromis niloticus
were studied using plastic buckets. Packed cell volume (peV) and histological study of gills and liver
were the tissue chemistry parameters investigated. The control buckets contained no effluent. Results
revealed increase in ESR (Erythrocyte Sedimentation Rate) decrease in hcamoglobin, red blood cell
and PCV Value of the fish exposed to all sub-lethal concentrations. Damage to liver and gills v.ere
noticed m some of the sub-lethal concentrations of all the effluent. Normal situation was observed In
all the examined tissues of fish from the control buckets. The African locust bean effluent is toxic on
0. niloticus and the lethal concentrations of African locust bean effluent deducted from the 96 hours
acute toxicity effect ranged from 3m!.
O. EKPO-.TTDE, O.A. FAGBEl\"RO & O.T. ADEBAYO
Department of Fisheries and Aquaculture Technology, Federal University of Technology, Akure
TOXICITY OF AFRJCA .."'JLOCUST BEA.."'JE"'FLlJE~T ON Oreochromis niloticus
Table 1. Water gualit~ of test solutions containing African locust bean effluents.
Concentration Day 1 Day 2 Day 3 Day 4
mVl Temp pH DO Temp pH DO Temp pH DO Temp pH DO
0 25 7.65 4.8 25 7.68 4.0 25 7.70 3.8 24 7.80 3.1
0 25 7.64 4.4 B 7.62 3.9 25 7.65 3.6 24 7.78 2.9
25 8.01 2.6 25 7.RS 0.6 26 7.75 1.0 24 7.69 0.9
25 7.52 1.7 25 7.55 0.7 25 7.74 1.1 25 7.62 0.9
2 25 7.65 1.4 24 7.63 0.9 25 7.70 0.6 24 7.70 0.6
2 25 7.70 1.2 25 7.65 0.7 24 7.72 0.5 24 7.68 0.6
3 25 7.52 1.0 25 7.53 0.5 23 7.43 0.5 25 7.80 OJ
3 24 7.65 0.8 24 7.65 0.6 23 7.66 0.4 25 7.77 0.1
4 25 7.60 0.9 24 7.73 0.4 25 7.75 0.5 25 7.74 0.2
4 25 8.02 0.7 25 7.75 0.3 25 7.73 0.4 25 7.72 0.1
5 25 7.66 0.3 24 8.01 0.1 24 7.78 0.1 25 8.01 0.1
5 25 7.82 0.2 24 7.98 0.1 25 7.90 0.1 25 8.01 0.1
RESULTS ANn DlSCUSSIOl"
Temperature and dissolved oxygen changed with the lime while pH Increased steadily. DIssolved
oxygen concentration declmed continuously, leading to stressful conditions on the fishes (Table 1).
Eileen et al.( 1991) also suggested that fish mortality tests arc affected by temperature, dissolved
oxygen concentration. pH and the duration of exposure. Fish mortality occurred in all conccntranons
(1, 2, 3, 4 or 5 ml) of the test toxicant as shown in Table 2. No fish died after 24 hours of exposure.
No mortality was observed on exposure of tilapia fingerlings to 1 ml or 2 ml of effluent throughout
the experiment however, mortality was found between 24 and 96 hours of exposure to 3 - 5 ml.
Generally. after the application of the effluent, must of the fishes gradually became imbalanced and
sank to the bottom within two hours of exposure. They later came to the surface and engulfed air by
projecting their snout above the water level. At this time they lost their swimming ability. Opercular
movements slowed down after one hour exposure.
Two replicates of the control treatment were also prepared by placing ten fingerlings in each of the
two containers filled with 100ml of water without the effluent.
This was followed by a 96-hour definitive test conducted using ten conico-cylindrical plastic
containers each filled with 1001111of water. each of the five stock solution varying concentrations
(Iml, 2ml, 3ml, 4ml and 5ml) of African locust bean effluent was introduced with a pipette into the
containers at 09.00 hours and the mixture was stirred. Stock solution and control treatments were
replicated twice. A total of 120 0. niloticus fingerlings (mean wt. 2. I5g) were used for this test. The
number of deal! and hvmg fingerlings in each plastic container was receded every 24 hours startmg at
O~.OO h. during which the behaviour of the fingerlings was monitored. Observations made include
mortality rate, lack of movement and lack of reaction to gentle prodding as described by Ward &
Parrish (1982). The percentage mortal ity 111 each concentration was determined and the LCso value
was dctermmed by the probit-logit transformation method. Water quality parameters such as
temperature, pH and dissolved oxygen (D02) concentration were determmed at 24 hour intervals using
standard methods described by MBA (1989) during 96-hour test. Immediately after death. gills and
liver of the fingerlings were excised upon dissecting the fish for histological examination according to
standard procedures. Blood samples were collected from two fingerlings from each of the treatment
media immediately after death by caudal puncture into 2.5m1 syringe already treated with ethylene
amino acetic acid (EDTA) to prevent coagulation. Packed cell volume (PCV), haemoglobin
concentration, leucocytes count and the erythrocyte count were estimated using various appropriate
laboratory methods as described by Blaxhall & Daisley (1973). Data collected were subjected to the
two-way analysis of variance (ANOV A) test (P = 0.05). Differences among the means were separated
by Duncan multiple range test (Duncan, 1955).
180
Table 3. Haematological value of Oreochromis niloticus fingerlings
The concentration of the African locust bean effluent used for the haematological test were 1ml, 2ml,
3rn1,4ml and 5mI for 96 hours, and theresults are presented in Table 3.
Concentration (ml/l)
The 96h effect of African locust bean effluent on Oreochromis niloticus fingerlings.
80
70
60
50
40
30
20
10
2 3 4 5 6 7
The exposure of tilapia to 5 and lOml of African locust bean effluent resulted in 20% and 70%
mortality, respectively, within 96 hours while exposure to 15, 20 and 25 ml resulted 111 50%, 60°-0,
80% and 100% mortality, respectively, within the exposure period. The 96 h LCso obtained
graphically (Fig I) was 8ml. The 96 hr LCso values are shown in Figure I. No mortality was observed
III tilapia exposed to 1 or 2 ml; on exposure to 3 - 5 ml, no fish died between 6 and 24 hours, however
mortality occurred between 48 and 96 hours of exposure. At 5mI concentration, the highest mortality
(30%) occurred between 48 and 96 hours of exposure. There was no mortality in the control
treatments and fish in this group did not show any abnormal behaviour.
Table 2. Mortality of Oreochromis niloticus exposed to African locust bean effluent for 96 hOUTS.
6 hours 24 hours 48 hours 72 hours 96 hours
Coneen lration % mortality Live Death Live Death Live Death Live Death Live Death
5lOl 20% 10 9 1 o 8 1 8
10ml 70% 8 2 7 I 7 6 1 3 3
15ml 70% 10 7 3 (, 1 6 3 3
20ml 90% 8 2 3 3 2 1
25ml 100% 8 2 3 5 1 2 0
Hb, RBC and PCV values of tilapia fingerlings exposed to 1,2,3,4 or 5 ml of the effluent decreased
significantly compared to those of the control fish while the RRC counts Increased at the end of the
exposure period compared to those recorded in the control. Exposure of 0. niloticus to sub-lethal
concentrations of African locust bean caused a significant decrease in rev, RBC and Hb of the fish.
Similar reduction in blood parameters was reported by Sarnpath et a/.( 1993) and Omoregic et
al.( 1994) when 0. niloticus W(I!) exposed to polluted environment under laboratory conditions. The
SIgnificant reduction in these parameters IS an indication of severe anaerrua caused by the effluent on
the exposed fish. Anaemic response could be due to the destruction of erythrocyte production
(Wintrobe,1978; OmoregIe,1995), haemodilution (Sampath et al.,1993), as well as the destruction of
intestinal cells (Gardner & Yerich,1970). Similarly, Rambhaskar & Rao (1990) noted that changes in
haernatological parameters due 10 unfavourable exogenous factors such as adverse water quality,
overstocking. and starvation are indices of ill health in cultivated fish. Ayotunde (1997) also reported
that stress factors result in changes in haematological characteristics in fish. It is concluded that some
parameters, and organs are affected negatively by increasing concentrations of the effluent and this
will guide in policy formulations against water pollution.
The gills and liver were examined lu assess the toxicological effect of the African locust bean
effluent on them. The various organs were exposed to different level of concentrations of the toxicants
(I, 2, 3. 4 or 5 ml). Mucus accumulation was observed on body surfaces and gill filaments of dead
fish. Examination of the gills of untreated (control) 0. niloticus fingerlings revealed a normal gill
filament consisting of primary lamellae with its arrays of delicate secondary lamellae, pnmary
epithelium and secondary epithelium covering the primary and secondary lamellae respectively and
no vacuolation, There was little ur no discernible change in gill structure of 0. niloticus fingerlings
exposed to low concentrations (1 and 2 ml) of the effluent. At higher concentrations however (3 and 5
1111) plates 10 and 12. the gill structure showed detachment of the epithelial cells in both the primary
and secondary lamellae, there was also vacuolation of the filament and influx of cells into the tissue
which resulted into degeneration of the tissue. Histological studies revealed that low concentranons
(1-2 ml) of the locust bean effluent did not cause any histological damage to 0. niloticus Ever. In
higher concentration (3-5 ml), the liver exhibited histological changes as indicated by loss of nuclei in
some hepatocytes, which also did not show distinct cytoplasmic boundaries as seen in the control fish
liver. There was space formation in the parenchyma tissue and the nuclear cells had thick dark look
(pykuosis) as seen in plate 3-5. The cellular arrangement of the liver cells was also distorted. At some
places, necrotic woes were observed where complete cell death was evident. Infiltration was also
clearly evident. Lesions were more prominent in the central zone of the liver than in the periphery.
Histological examination revealed that from the treated fish species, the gills were swollen and the
lamellae were extensively fused and congested with blood. Similar observations were made by
Onwumere (1986), who expressed that the histology of Iiver and gills of 0. niloticus fingerlings
exposed to 30, 40 and 50% effluent from the NNPC Refmery at Kaduna showed that the gills were
swollen and bulged the opercula. Aderiyc (1998) also stated that the gill structure of 0. niloticus
treated with petrol and engine oil mixture was fused together and that there was extensive hyperplasia
and separation of the epithelial layer from the supportive tissues. In the liver, degeneration vacuole
formauon and irregular nuclear was noted and these was observed in newborn guppies, Lebistes
reticulates (Crandall & Goodnight, 1962), disintegration and necrosis in common carp, Cyprinus
carpio (Wong et ul.,1977) and vacuolation, necrosis and appearance of some globular bodies In
Puntius eonchonius (Kumar & Panl,1981) due to zinc toxicity were reported.
Concentration of test media ESR PCV RBC Hb
(mm2 (%) {mm3} (g(mmll_
Control 4.0 30 15300000 26
1ml 5.0 28 1470000 24
2rnl 5.5 28 1500000 26
3 ml 6.0 25 135000CO 23
4rnl 6.5 24 1348000 23
5 m1 7.5 21 1160000 21
30
182
REFER..I!:NCI!:S
Aderiyc, R.K (199R) Studie« on the toxicological effects of petrol and engme oil mixture on African
catfish, Clurtus garieptnus (Burchell 1822) fingerlings. ;\,1. Agnc 1echo °1besis. Federal
Umversity of Technology. Akure. 65pp.
Ahmed. I.M., Farouq. A.A. Yerirna, '\1.B. & Bello, A.M.(2005) Acute toxicuy ot'watei extract of the
bark of African locust bean on tilapia (Oreochromis niloticusi. Pp7. 111 Book of Abstracts 41 st
Annual Conference of Science Association cf Nigeria, Apnl 25-29, 2005. Lsmanu Danfodiyo
University, Sokoto, Nrgeria.
Annune. P.J\. & AJlke. S.U.(1999) Acute Toxicity and gill morphology of Oreochromis 1IIIutl(7/\
exposed to Roger". Journal of Aquatic Sciences 14: 1-4.
APIlA, AWWA & WprF (American Puhlic Health ASSOCiation, American Public Health
Assoctauon, Ameucan Water Works Association and Water Pollution Control federal) (1989)
Standard Methods for the Examination of Water and Wastewater. 17111Edition. APHA,
Washington, DC. 139l pp.
Ayotunde E.O.(l997) Ilaernatological ami serological characteristics or He/era/is niloticus (Cuvrer
1829): Osteoglossidae, M.Agric.Tcch. Thesis. Federal University of Technology. Akure. 82pp.
Blaxhall, P.C. & Daisley, K.W.(1973) Routine haernatological methods for use with fish blood,
Journal ofl-ish BIOlogy, 5:771-781.
Crandall, c.A. & Goodnight. C.1.(1962) Effects of sub-lethal concentrations of several toxrcants of
growth of the common guppy, Lebistes reticuuues. Limnology and Oceanography 7: 233-2)<)
Eileen, M.P .. Wilham, R \V., James. W.A. & Guthrie vV.P.(IY91) loxicity of chelated copper to
juverule-, of red drum .. Sciuenops ocellatus. Journal of the World Aquaculture Society 3: 101-108.
Gardner, G.R. Ix Yevich, P.P.( 1970) Histological and haematological responses of an estuarine teleost
to cadmium Journal of Fishenes Research Board of Canada 27: 2185-2196.
Grizzle. J.M.(1977l l laematological changes in fingerlings channel catfish exposed to malachite
green. Progressive FIsh Cultunst 39: 90-93.
Krous, S.R., Bluer, V.S. & Mead, T.L( 1982) Effects of acclimation of nitrite movement across the
gill epithelia of rainbow trout. Progressive Fish Culturist 44: 126-130
Kumar, S. & Pant, S.c.( 1981) Histopathological effects of acutely toxic levels of Cu and Zn on !,'111s.
liver and kidney of Puntius conchonius. Indian Journal of Experimental Biology 19(2): 191-194.
Oladimeji, A.A.(l987) Impacts of oil pollution on Nigerian fishing industry. Nigerian Journal Applied
Fisheries and Hydrohiology 2: HI-YO
Omoregie, E.(1995): Effects of petrol on the Nile tilapia and its helminth infection. Ph.D. Thesis
University of Jos, Jos Nigeria. 152pp.
Omoregie, E., Eseyin, T.G. & Ofojekwu , P.C.(1994) Chronic effects or lorrnalin on erythrocyte
counts anti plasma glucose nlNile tilapia, Oreochromis niloticus. Asian Frshencs Science 7: 1-6.
Ornorcgic, E .. IJfodike. E.R.C. & Keke, l.R.(l990) Tissue chemistry of Oreochromis niloticus
exposed to sub-lethal concentrations of Gammalin 20 and Actellic 25 Ec: Journal of Aquatic
SCiences 5: 33-36
Onuoha, G.c. & ]\wadukwc. F.O.( 1990) Influence of liquid petroleum refinery effluent on the
hatching succcxs of Cluriu» gariepinus (African mudfish) e)!!-!:;.Journal of Environment and
Ecology X14. 1201-1206.
Onwumere, H.Cd 1986) Toxicity of treated eft1uent from NNPC Refinery, Kaduna to ulapia
tOreochrouus mloticusi. M.Sc. Thesis, Ahmadu Delio Umversuy, Zaria.
Oram, R.F., Hummer, P.l. & Smoot, R.C.(1976) Biologyof livmg systems. Charles. E. Merrit
Publishing Company, Columbus, Ohio.
Rambhas Kar, B. & Rao, K.S.( 19<)0) Use of haernatological parameters as diagnosnc tools In
deterrmrung the health of rmlkfish, ebanos chanos (Forsskali) in brackish water culture.
Aquaculture and Fishenes Management 21: 125-129.
Sampath, K., Velarnnial, S., Kennedy, 1.1. & James, R.( 1993) Haematological changes 'on
Oreochromi ...mossumhicus as a function of exposure penod and sub-lethal levels.
Sikoki. F.D. & Kolo, R.J.(1993) Perspectives in water pollution and their implication for conservation
of aquatic resources. Pp. 184-190, In: Proceedings of the National Conference on Conservation
of Aquatic Resources. Egborge, A.B.M., Omuloyin. OJ., Olojcdc, A. & Manu, S.A. (eds.)
NARESCON Publishers, Abuja, Nigeria. 31Spp,
Wintrobe, M.M.(19?g) Clinical Haematology. H. Kiimpton, London, UK. 488pp.
Wong, M.H, Luck, K.C. & Choi, K Y (1977) The effects of Zn and Cu salts on Cyrinus carpio and
Ctenopharvngodon idellus. Acta-Anatornica ~.1:450-454.


Toxicity of African locust bean effluent on Oreochromis niloticus

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Toxicity of African locust bean effluent on Oreochromis niloticus

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