Co esponding au ho : Allison Theodo e A hanasius.
Copy igh © 2025 Au ho (s) e ain he copy igh o his a icle. This a icle is published unde he e ms o he C ea i e Commons A ibu ion License 4.0.
His ological based sub-ch onic oxici y es ing o a ge hea y me als in esh wa e ;
Using he His o-mo phome y o Kidney o Cop odon Zilli
Allison Theodo e A hanasius 1, * and Ogoun Timipa Richa d 2
1 Depa men o Ana omy, Facul y o Basic Medical Sciences, College o Heal h Sciences, Uni e si y o Po Ha cou , Nige ia.
2 Depa men o Ana omy, Facul y o Basic Medical Sciences, Bayelsa Medical Uni e si y, Nige ia.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2637-2646
Publica ion his o y: Recei ed on 28 Ma ch 2025; e ised on 09 May 2025; accep ed on 11 May 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.26.2.1688
Abs ac
This his ology based oxicological s udy is aimed o ad ance he his opa hological cha ac e is ics o ish kidney exposed
o some known c ude oil spill hea y me als. Based on li e a u e e iew on he en i onmen al c ude oil spill bu dens o
Nige Del a egion o Nige ia, he ollowing a ge chemicals (TC) we e selec ed o he s udy: Cadmium (Cd), Ch omium
(C ), Coppe (Cu), Nickel (Ni) and Lead (Pb). The expe imen al ype is a sub-ch onici y ish exposu e es ing o wen y-
eigh (28) days du a ion. The expe imen al se up in ol es wen y- wo pieces (22) o wen y-li e (20l) anks o ou
(4) eplica e anks pe a ge chemical (TC) con aining en (10) ju enile s udy ish pe ank in a semi-s a ic ank es ing
sys em. TC exposu e concen a ions abo e maximum allowable oxican concen a ion (MATC) o esh wa e we e
used o he s udy. Quali y his ologically esul s showed al e a ion which we e consis en wi h hea y me al exposu e
s udies. Semi-quan i a i e assessmen showed signi ican di e ence be ween con ol and expe imen al kidney o gan
his opa hological indices. The s udy u he alida es MATC esh wa e guideline use o oxici y ce i ica ion in C , Cd
and Ni exposu e.
Keywo ds: His ology; Toxici y; Toxicology; Sub-Ch onic; Hea y Me als and Fish Kidney
1. In oduc ion
This s udy is aimed o simula e he ecological e ec o exposu e o some known hea y me als om c ude oil spill. I is
a labo a o y based sub-ch onic oxici y es ing o hea y me al ea ed esh wa e using he kidney o a bio-indica o
ish species. Toxici y es s a e used o expose es o ganisms o a medium—wa e , sedimen , o soil—and e alua e he
e ec s o con amina ion on beha io al, physiological, biochemical and his ological end poin s. The goal o oxici y
assessmen is o iden i y ad e se e ec s o a subs ance. Ad e se e ec s depend on wo main ac o s (O oboni, 1991);
•Dosage: has o do wi h du a ion and concen a ion o exposu e. Bo h la ge single exposu es (acu e) and
con inuous small exposu es (ch onic) a e s udied.
•Rou e o exposu e: The ou es o exposu e a e Inges ion (o al), inhala ion o skin abso p ion (de mal).
•O he ac o s a e species, age, sex, heal h, en i onmen and indi idual cha ac e is ics.
Based on he measu ed end poin s, oxici y es ing di ided in o Acu e, Sub-Ch onic and Ch onic oxici y es ing (USEPA,
1994:
•Acu e oxici y es s: a e sho - e m es s ha measu e he e ec s o exposu e o ela i ely high concen a ions
o chemicals. The measu emen endpoin gene ally e lec s he ex en o le hali y (USEPA, 1994).
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• Sub-Ch onic Toxici y Tes ing: A oxici y es designed o in es iga e possible ad e se e ec occu ing as a esul
o con inuous o epea ed exposu e o se e al g oups o expe imen al animals o a se ies o concen a ion o
he es chemical o pa (no exceeding 10%) o hei li e span (ECETOC. 1986)
• Ch onic oxici y es s, on he o he hand, gene ally a e longe - e m es s ha measu e he e ec s o exposu e
o ela i ely lowe , less oxic concen a ions. Fo a ch onic oxici y es , he measu emen endpoin conce ns a
suble hal e ec (e.g., ep oduc ion, g ow h) o bo h le hali y and sub- le hal e ec (USEPA, 1994). Resul s can
be analyzed in se e al ways. One is simply by a di ec compa ison be ween pe cen e ec s occu ing in
o ganisms exposed o si e media and hose exposed o uncon amina ed media.
• O he app oaches o analysis de e mine he e ec concen a ion on i y pe cen (50%) o he sample (EC50)
and he lowes obse ed e ec i e concen a ion (LOEC), o he no obse ed e ec concen a ion (NOEC). The
e ec , “E” in he a o emen ioned ac onyms can be any o he ollowing end poin s: mo ali y, mo bidi y,
inhibi ion o g ow h and/ o ep oduc ion.
Hea y me al con amina ion o aqua ic ecosys em has a ac ed g ea a en ion o esea che s o e he las ew decades
(Fa ombi e al., 2007). Me al in he aqua ic en i onmen a e bioaccumula ed by o ganisms ei he passi ely om wa e
o by acili a ed up ake. Excess me al concen a ion in an o ganism mus be ac i ely exc e ed, compa men alized in
cells o issues, o me abolically immobilized. Some me al escape all hese ac ions causing oxic and o he ad e se
e ec s (Chapman e al., 1996; Rand e al., 1985). Some hea y me als, Ch omium (C ), cadmium (Cd), coppe (Cu), lead
(Pb), nickel (Ni) and zinc (Zn), a e ound in he lis o o ganic and ino ganic haza dous pollu an s which is p epa ed by
he Uni ed S a es En i onmen al P o ec ion Agency (USEPA) (Aka and Tunali, 2005). Howe e , i is impo an o no e
ha mos essen ial me als may be oxic when p esen in concen a ions ha a e in excess o he guideline le els (Agbozu
e al., 2007). Acco ding o Robe s (1989 and 2001) Cu, Pb, me cu y (Hg), Zn, C , manganese (Mn) and i on (Fe) a e he
mos common cause o me al poisoning. Ja up (2003) documen ed ha he main h ea o human heal h om hea y
me als a e associa ed wi h exposu e o Pb, Cd, Hg, and a senic (As) (a senic is a me alloid bu is usually classi ied as a
hea y me al).
Ge ha d (2004) es ablished ha bioindica o s a e o ganisms o communi ies o o ganisms which eac ions a e
obse ed ep esen a i ely o e alua e a si ua ion, gi ing clues o he condi ion o he whole ecosys em; Ge ha d also
indica e ha bioindica o s a e species eac ing o an h opogenical e ec s on he en i onmen , concluding ha a
biological indica o would be: a species o g oup o species ha eadily e lec s he abio ic o bio ic s a e o an
en i onmen , ep esen s he impac o en i onmen al change on a habi a , communi y o ecosys em o is indica i e o
he di e si y o a subse o axa o he whole di e si y wi hin an a ea. In his sense, he p ima y ole o ecological
indica o s is o measu e he esponse o he ecosys em o an h opogenic dis u bances (Niemi and McDonald, 2004).
Kidneys a e i al o gans in he main enance o pH and olume o body luids as well as e y h opoiesis (Iqbal e al., 2004).
In ish, hey main ain a wa e and elec oly e balance in o de o p o ide a s able in e nal en i onmen . Due o hese
p ope ies, he kidneys a e also an ou s anding indica o o possible pollu ion and en i onmen al s ess in he icini y
o he ish (Hin on and Lau en, 1990). Fish kidney neph ons consis o collec ing duc s, p oximal and dis al ubules, and
enal co puscle. Mo eo e , he in e s i ial s oma o ish kidney includes hema opoie ic issue, which con ains
hema opoie ic s em cells (HSC), blood cells, enal p ogeni o s, and o he mesenchymal cell ypes (Mo amedi, e al.,
2020). The kidney o mammals canno o m new neph ons o eplace he los ones when i is damaged, and i can epai
only he p oximal ubules and glome uli pa ially h ough bone ma ow cells (Mo amedi e al., 2019) o in a enal
mesenchymal cells (Lin e al., 2005; Osa une e al., 2006). Howe e , a ious ish species, including ainbow ou , ilapia,
ca ish, zeb a ish, omcod, and he Ho muz killi ish Aphaniops ho muzensis a e known o egene a e hei kidneys
((Mo amedi, e al., 2020). Fish kidneys a e conside ed as an impo an i al o gan o use as a bioma ke in de e mining
he heal h o ish in he eshwa e ecosys em (Be ne e al., 1999). The issues and o gans which a e in con ac wi h he
o eign compounds ha e he po en ial o su e he mos and cause a damage when exposed o highe concen a ions o
chemical pollu an s (Timb ell, 1991).
His ology is he s udy o he s uc u e o no mal issue while his opa hology is he s udy o he s uc u e o abno mal,
diseased issue. Acco ding o Hin on and Lau en (1990) his ological al e a ions a e he o e all esul o un a o able
physiological and biochemical al e a ions in an o ganism. They may be used o o esee he e ec s o al e a ions on
p ocesses o ac i i ies such as p eda o a oidance, g ow h, ep oduc ion, and popula ion s abili y ha occu a highe
le els o biological o ganiza ion (Meye s and Hend icks, 1985). Nega i e e ec s as a esul o oxici y gene ally begin a
he cellula o issue le el long be o e he ob ious signs such as beha io al changes, loss o appe i e, discolo a ion, o
dea h occu . This is whe e his ology could be a use ul ool in oxici y s udies. His opa hology was in oduced in s udies
whe e mo e de ailed in o ma ion was needed o u he desc ip ion o sc eening o ea ly p eneoplas ic changes
(Wes e e al., 2002). His opa hological cha ac e is ics o speci ic o gans exp ess condi ion and ep esen ime
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2637-2646
2639
in eg a ed endogenous and exogenous impac s on he o ganism s emming om al e a ions a lowe le els o biological
o ganiza ion (Hin on and Lau en, 1990; Swee e a ., 1996).
Di ec exposu es o a ious oxic pollu an s may esul in abno mali ies and pa hological al e a ions in ish (Be ne e
al., 1999). The e o e, o de e mine and simula e he e ec s o some a ge hea y me als exposed o Cop odon Zillii, a
his ology-based ish heal h assessmen p o ocol was used o e alua e he bio-indica o ish, using he kidney as a
his ologic bioma ke
2. Ma e ials and me hod
2.1. S udy Loca ion
The ish oxici y es ing was done in a We labo a o y o he Depa men o Aquacul u e and Fishe ies Managemen ,
Facul y o Ag icul u e, Uni e si y o Benin, Nige ia. The depa men unde akes high le el human esou ce de elopmen
h ough esea ch, aining and echnology ans e o ensu e sus ainable de elopmen o o es esou ces as well as he
en i onmen al and social impac on hese esou ces.
2.2. S udy Specie
2.2.1. Fish selec ion
EROCIPS (Eme gency Response o coas al Oil, Chemical and Ine Pollu ion om Shipping), (2006), “P o ocol o
Selec ion o Sen inel Species” was basically used o selec he app op ia e sen inel specie o his s udy. The esiden
ish species, Cop odon Zillii because i is a widely s udied ish species (Ha ey e al., 1999; Pie apiana e al., 2002;
Budzinski e al., 2004; EROCIPS, 2006; Ma igo´mez e al., 2006; Ma ı´nez-Go´mez e al., 2006; Joly-Tu quin e al., 2009),
and i is a esiden o he Nige Del a egion o Nige ia. This egion is c ude oil p oduc ion hub o Nige ia, wi h he highes
le el o c ude oil spills and o he c ude oil ela ed en i onmen al c imes.
Figu e 1 Pic u e o C. zillii
2.2.2. Fish Bioda a
The chosen bio-indica o ish specie o his s udy is Tilapia zillii, now known as Cop odon Zillii. The ish specie is a
esiden o he Nige Del a egion o Nige ia. Cop odon Zillii has a maximum leng h o 40cm (SL) and a maximum
published weigh o 300 g ams wi h a o al o 13 o 16 Do sal spines (GISD, 2019). The non-b eeding colou a ion o C.
zillii is da k oli e on op and ligh oli e o yellow-b own on he sides, o en wi h an i idescen blue sheen. Lips a e b igh
g een and he ches is pinkish. Six o se en da k e ical ba s c oss wo ho izon al s ipes on he body and caudal
peduncle. Fins a e oli aceous, co e ed in yellow spo s wi h he do sal and anal ins displaying an ou line o a hin o ange
band. Caudal in o en g ey wi h pale in e s ices wi h do s co e ing he en i e in. Adul s display a black spo ou lined in
yellow. C. zillii om 2 o 14cm (SL) ha e an en i ely yellow o g ey caudal in wi h no do s, de eloping a g eyish caudal
in wi h do s wi h inc easing size. Spawning colo a ion is shiny da k g een on op and sides, ed and black on he h oa
and belly, and ob ious e ical bands on he sides. Heads u n da k blue o black wi h blue-g een spo s. Eggs a e g een
o oli e g een, s icky, 1-2 mm in diame e ; ela i ely smalle han eggs o o he cichlids (FishBase, 2008; GISD, 2019).
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2637-2646
2640
2.2.3. Fish Sou ce
The ish o oxici y es ing was acqui ed om Nige ia Ins i u ion o Oceanog aphy and Ma i ime Resea ch (NIORMR)
in Sapele Del a S a e, Nige ia. The Ins i u e is in ol ed in a me s' ield es and incuba ion alida ed esea ch esul s
and echnologies. The es ish was posi i ely iden i ied by a axonomis om NIOMR, o be he igh species o he
s udy. The es ish used was disease- ee and appea heal hy, beha e no mally, eed well, and had low mo ali y in
cul u es, du ing holding, and in es con ol. (USEPA, 2002).
2.2.4. Tes Fish Guidelines
Ju eniles o Cop odon Zillii we e chosen o he s udy in andem wi h ISO,1994 ecommenda ion o ch onic oxici y
s udy (ISO, 1994; CEPA, 1999). Young o ganisms a e o en mo e sensi i e o oxican s han (a e) adul s. Fo his eason,
he use o ea ly li e s ages, such as ju enile is equi ed o all es s (ISO, 1994; CEPA, 1999). Fish we e app oxima ely
he same age and we e go en om he same sou ce. Since age may a ec he esul s o he es s, his would enhance
he alue and compa abili y o he da a i he same species in he same li e s ages we e used h oughou a moni o ing
p og am a a gi en acili y (ISO, 1994; CEPA, 1999).
2.3. S udy Design
2.3.1. S udy Guideline
In e na ional S anda diza ion O ganiza ion (ISO) es guideline s anda d, ISO 10229:1994 – “Ch onic Toxici y Tes o
Fish Wa e quali y”, was used o he labo a o y s udy (ISO, 1994). The s anda d speci ied he me hod o he
de e mina ion o he long- e m oxici y o subs ances (pu e chemicals, mix u es, was ewa e e c.). This S anda d allows
o he use o a semi-s a ic me hod. The endpoin esponse was a measu e o he mo phological changes o es ish
exposed o a es subs ance o a pe iod o 14 o 28 days. The s anda d pe mi s o adap his me hod o use wi h a wide
a ie y o eshwa e , ma ine and b ackish wa e ish wi h app op ia e modi ica ions in es condi ions ( empe a u e,
ood, ish ma king echnique).
2.3.2. Expe imen al Se -up
Sub-Ch onic Toxici y s udies we e done in Depa men o Aquacul u e and Fishe ies Managemen , Facul y o
Ag icul u e, Uni e si y o Benin. The expe imen al du a ion was 28 days. Twen y wo pieces (22) o wen y li e s (20L)
anks we e acqui ed o he expe imen . Fou eplica e anks pe Ta ge Chemical (TC) we e labeled and used o he
g oup ea ed. Hence, i e TC o Coppe , Cadmium, Ch omium, Lead and Nickel, a o al o Twen y (20) anks we e se up
o he expe imen .
Figu e 2 Expe imen al se -up in he We -labo a o y o he Depa men o Ag icul u e Uni e si y o Benin
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2641
2.3.3. Tes Chemical/Concen a ion
Ta ge Chemicals (TC), Cadmium (Cd0, Ch omium (C ), Coppe (Cu), Nickel (Ni) and Lead (Pb) we e chosen because o
hei ca diogenic quo ien and p esence in c ude oil. A s anda d s ock solu ion o 100mg/L o he TC (Cd, C , Cu, Ni and
Pb) was p epa ed om analy ical g ade me allic sal s o Cadmium Chlo ide (CdCl2), Lead II Ni a e (Pb(NO3)2), Nickel
II Sulpha e (NiSO4.(H2O)6), Coppe Sulpha e (CuSO4) and Po assium Dich oma e (K2C 2O7). S ock solu ion was
p epa ed wi h de-ionized wa e in I-L olume ic. The maximum accep able oxican concen a ion (MATC) o
es ua ine esh wa e was se as he concen a ion o s ock solu ions o he labo a o y es ing (Gheo ghe e al., 2017).
Two concen a ions below and abo e he es ima ed MATC we e used o he es ing (Gheo ghe e al., 2017).
Table 1 The esul s o TC doses applied o he ish oxici y s udy
Ta ge Chemicals (TC)
MATC (mg/L)
FED: Dose abo e MATC (mg/L)
FED: Dose below MATC (mg/L)
C1
C2
C3
C4
Cadmium (Cd)
0.00001
0.1
0.01
0.0001
0.0001
Ch omium (C )
0.001
10.0
1.0
0.01
0.001
Coppe (Cu)
0.002
20.0
2.0
0.02
0.002
Nickel (Ni)
0.025
25.00
2.5
0.05
0.025
Lead (Pb)
0.001
10.0
1.0
0.01
0.001
Key: TC = Ta ge Chemicals; FED = Fish Exposu e Dose; C1, C2, C3 and, C4 = Dose Concen a ions o Hea y me al applied pe Fish Tank; MATA =
CCME, 2001
2.3.4. Tes Chambe
Twen y (20) li e s o chemically ine essels (plas ic Tanks) we e used o he s udy. Each anked was s ocked wi h 25
ish, a s ock capaci y ha was enough o allow o p ope g ow h and main enance o dissol ed oxygen concen a ion.
This is in compliance wi h he guideline loading a e c i e ia (OECD, 2013). The es chambe s was andomly posi ioned
in he es a ea and shielded om unwan ed dis u bance. The es was ca ied ou wi hou adjus men o pH.
Ne e heless, whe e he e is e idence o ma ked changed in he pH o he Tank wa e a e addi ion o he es
subs ance, he es would be epea ed, adjus ing he pH o he s ock solu ion o he ank wa e be o e addi ion o he
es subs ance. The PH adjus men will be made (p e e ably wi h HC1 o Na0H) in such a way ha he s ock solu ion
concen a ion is no changed o any signi ican ex en and ha no chemical eac ion o physical p ecipi a ion o he es
subs ance is caused (ISO, 1994; CEPA, 2004; Gheo ghe e al., 2017).
2.3.5. Tes Solu ion Condi ions
The es solu ion is esh wa e , which was used o simula ed esh su ace wa e . Wa e empe a u e was main ained
be ween 20OC o 25OC (ISO, 1994; CEPA, 2004). The empe a u e o he es solu ions was measu ed by placing a
he mome e di ec ly in o he es solu ions. Tempe a u e was eco ded con inuously in a leas one essel du ing he
du a ion o each. DO concen a ion was main ained a no less han 60% o he maximum ai sa u a ion alue
h oughou he es . DO and pH was checked a he beginning o he es and daily, h oughou he es pe iod. Ligh
quali y was se a ambien labo a o y illumina ion. Pho ope iod was se a a minimum o a ion o 12 hou s ligh o
12hou s da k, wi h a ligh in ensi y main ained a 10 o 20 μE/m2/s. Feeding was a leas once daily, he quan i y o
ood being kep cons an and ela ed o he ini ial ish weigh , a leas 2% body weigh (ISO, 1994; CEPA, 2004).
2.3.6. Validi y o Tes
• Fo he condi ions o alidi y, ISO (1994) and OECD (2013) condi ions o he alidi y o es we e adop ed o
his s udy:
• The mo ali y in he con ols should no exceed 10% a he end o es .
• The dissol ed oxygen concen a ion should be a leas 60% o he ai sa u a ion alue h oughou he es
• In semi-s a ic p ocedu es, ae a ion can be used, p o ided i does no lead o a signi ican loss o es subs ance
• The e should be e idence ha he concen a ion o he subs ance being es ed has been sa is ac o ily
main ained (i should be a leas 80% o he nominal concen a ion) o e he es pe iod. The esul s should be
based on measu ed concen a ion i he de ia ion om he nominal concen a ion is g ea e han 20%
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2.4. His ological E alua ion
A he end o he expe imen , 5 ishs om each ank was sac i iced and he a ge o gan o gills we e excised and
p ese ed in 10% bu e ed o malin solu ion and sen o he labo a o y o issue p epa a ion. Tissue p epa a ion was
done in acco dance o D u y and Walling on (1980). The ollowing his ological e alua ions we e made:
2.4.1. Quali a i e Analysis
A Ligh mic oscopy (Olympus BH2) was used o iden i y and in e p e (his ological desc ip ion) issue slides and
mic og aph specimens a 40X, X100 and X400 magni ica ion. The pe cen age p e alence o issue his opa hology was
no ed. His ological indings in he p epa ed slides om Fish Exposed Dosage (FED) o TCs and con ol we e no ed.
2.4.2. Semi-quan i a i e analysis
A s anda d semi-quan i a i e his ological assessmen acco ding o he modi ied p o ocol o Be ne e al. (1999)
modi ied by Van Dyk e al. (2009) was done o quan i y he his opa hological al e a ions obse ed in he gill o his ology
slides. This assessmen is use ul because i can be applied o any o gan as well as allowing o s anda dized
quan i ica ion o esul s. This assessmen allows o he compa ison o obse ed his opa hologic lesions be ween he
di e en Fish Exposu e Dosage (FED) o TCs and con ols (USEPA, 1994).
3. Resul s
3.1. Quali a i e His ological Assessmen
Figu e 3 Pho omic og aphs (H&E:400X). (A) No mal Kidney show Renal Co puscles (RC) and Renal Tubules (RT) (B):
Showing mul i ocal Melano Mac ophage Cen es (MMC) and di use In e s i ial Haemo hage (IH) (C): Showing Mul i-
ocal MMC wi h ma ked s uc u al al e a ion – ha dening wi h a ophy o enal co puscles and enal ubules
3.2. Semi-Quan i a i e His opa hological Assessmen
Below (Table 2) is he kidney his opa hologic o gan index (Io g) esul collec ed wi h a sco e cha quan i ied by Be ne
e al. (1999) p o ocol (modi ied by Van Dyk e al., (2009a), showed ha : Li e Io g o Cd1-Cd3, Cu3, Ni3, Pb2 and Pb3
we e abo e con ol; Kidney Io g o Cd1, Cd2, C 1, C 2, Cu3, Ni3, Pb2 and Pb3 we e abo e con ol. Gills Io g o Cd1, Cd2,
C 1-C 3, Cu3, Ni3, Pb2 and Pb3 we e abo e con ol. Mean ish indices esul showed ha Cd1, Cd2, C 1, C 2, Cu3, Ni3,
Pb2 and Pb3 we e abo e he con ol alues
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Table 2 The mean O gan and ish Indices o cul u ed ish lesions
TC
FEC
Kidney Mean Io g
Con
1.0
Cd
Cd1
13.4
Cd2
8.6
Cd3
0.6
Cd4
0.2
C
C 1
11.0
C 2
10.4
C 3
0.4
C 4
0.4
Cu
Cu1
Nil
Cu2
Nil
Cu3
4.6
Cu4
1.0
Ni
Ni1
Nil
Ni2
Nil
Ni3
2.6
Ni4
0.4
Pb
Pb1
Nil
Pb2
9.6
Pb3
1.2
Pb4
0.2
Key: Cd1, Cd2…C 1, C 2… e c = Concen a ions o Hea y me al applied pe Fish Tank; Conc = Concen a ion: ed = alues abo e he con ol alues;
FEC = Fish Exposu e Concen a ion; Io g = O gan Index; Nil – Fish in he es chambe did no su i e he du a ion o he expe imen
Table 3 Kidney Pai ed Sample T Tes o he Fish Toxici y S udy, compa ing he mean O gan Indices (Io g) o ish om
he di e en FED anks wi h he con ol ish
Pai ed Di e ences
D
Sig. (2- ailed)
Mean
S d. De ia ion
S d. E o Mean
95% Con idence In e al
Lowe
Uppe
Cd
Con - Cd1
-12.800
6.140
2.746
-20.424
-5.176
-4.661
4
.010
Con - Cd2
-8.000
2.915
1.304
-11.620
-4.380
-6.136
4
.004
Con - Cd3
0.000
1.000
.447
-1.242
1.242
0.000
4
1.000
Con - Cd3
.400
1.140
.510
-1.016
1.816
.784
4
.477
C
Con - C 1
-10.400
7.021
3.140
-19.118
-1.682
-3.312
4
.030
Con - C 2
-9.800
4.658
2.083
-15.584
-4.016
-4.704
4
.009
Con - C 3
.200
.837
.374
-.839
1.239
.535
4
.621
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2637-2646
2644
Con - C 4
.200
1.304
.583
-1.419
1.819
.343
4
.749
Cu
Con - Cu1
.600
.894
.400
-.511
1.711
1.500
4
.208
Con - Cu2
.600
.894
.400
-.511
1.711
1.500
4
.208
Con - Cu3
-4.000
2.345
1.049
-6.912
-1.088
-3.814
4
.019
Con - Cu4
-.400
1.140
.510
-1.816
1.016
-.784
4
.477
Ni
Con - Ni1
.600
.894
.400
-.511
1.711
1.500
4
.208
Con - Ni2
.600
.894
.400
-.511
1.711
1.500
4
.208
Con - Ni3
-2.000
1.871
.840
-4.323
.323
-2.390
4
.075
Con - Ni4
.200
1.304
.583
-1.419
1.819
.343
4
.749
Pb
Con - Pb1
.600
.894
.400
-.511
1.711
1.500
4
.208
Con - Pb2
-9.000
5.657
2.530
-16.024
-1.976
-3.558
4
.024
Con - Pb3
-.600
.547
.245
-1.280
.080
-2.449
4
.070
Con - Pb4
.400
1.140
.510
-1.016
1.816
.784
4
.477
Key: TC = Toxican Conc,; FEC = Fish Exposu e Conc. NA = No Applica ion (Fish g oup all died); RED = alues abo e he con ol alues
4. Discussion
The kidney his olological al e a ions include: Di use In e s i ial Oedema; gene alized ascula up u e wi h In e s i ial
Haemo hage (IH); Melano-Mac ophage Cen e; Vacuola ion; Di use Coagula i e Nec osis; Renal Tubula Hype ophy;
and Nec o ic Foci. This s udy indings was consis en wi h his opa hological al e a ions ound in ish exposed o a ious
con aminan s which include: ubula epi helium and glome ulus (Teh e al.,1997); dila ion o Bowman’s space
(Ji aungkoo skul e al., 2002); ubula nec osis, desquama ion and acuola ion o ubula epi helial cells (O iz e al.,
2003); degene a ion o glome uli and inc eased in in a ubula hema opoie ic issue (Iqbal e al., 2004); and
degene a ion o glome ulus, hype ophied cells and na owing o he ubula lumen (Cengiz, 2006).
The semi-quan i a i e analysis showed ha he TC concen a ions abo e MATC had wo se kidney o gan his opa hologic
indices (Io g) han con ol. This alida es he guideline MATC o esh wa e aqua ic li e (ECETOC,2002) which
p oposes dele e ious ecological exposu e e ec beyond he allowable en i onmen al concen a ion o he TC. Though
concen a ions o Cu3, Cu4 & Pb3 caused haza dous e ec e en a concen a ions lowe ha he MATC. This migh no
be un ela ed o he haza dous quo ien o he me als, bu ne e heless needs u he in es iga ion. Fu he mo e, he e
was signi ican di e ence (P<0.005) o he kidney his opa hologic o gan indices (Io g) be ween he con ol and TC
concen a ions o Cd1, Cd2, Cu3 and Pb2.This u he alida es he MATC guideline o esh wa e aqua ic li e
(ECETOC,2002). Cu3, which is a Cu concen a ion below MATC, showed haza dous exposu e e ec which was
inconsis en wi h MATC guideline. This migh be due Cu oxic a ini y o kidney and o Haza dous Quo ien in esh
wa e bodies. Ne e heless, his ac needs u he in es iga ion
5. Conclusion
This s udy was oxicologically ele an . The s udy was able o demons a e ha hea y me al oxici y can be simula ed
in a labo a o y se ing. I has u he p o en ha he a ge chemicals concen a ion abo e he guideline le els o
egula o y s anda ds o MATC we e haza dous o he bio-indica o ish. Though he e is need o u he in es iga e he
pha macokine ics behind he haza dous exposu e e ec s caused by he concen a ions o Cu and Pb ha we e below
MATC guideline, which was inconsis en wi h MATC egula o y p o ision. The s udy has also p o en ha
his opa hologic o gan indexing is an explo able en i onmen al ool o his ological based oxici y ce i ica ion
Compliance wi h e hical s anda ds
Disclosu e o con lic o in e es
No con lic o in e es o be disclosed.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2637-2646
2645
S a emen o e hical app o al
E hical app o al was ob ained.
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