Research Article ISSN 2250-0480 Vol 3/Issue 2/Apr-Jun 2013

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Lie Science !io "echnolo#$


HISTOPATHOLOGY CHANGES IN FRESH WATER FISH CLARIAS
BATRACHUS(Linn.) EXPOSED TO MERCURY AND CADMIUM

J. SELVANATHAN
1
, S.VINCENT
2
AND A. NIRMALA
1

1
Department of Biotechnology, Vinayaka Missions University, Aarupadi Veedu
Institute of Technology, hennai!"#$1#%, Tamilnadu, India&
'
Associate (rofessor, Department of Advanced )oology and *ealth Biotechnology,
+oyola ollege, hennai!$%& Tamil ,adu,India&


ASTRACT

"he release o hea%$ &etals into the a'uatic en%iron&ent causes (ater pollution pro)le&s )ecause o their
to*icit$+ persistence an, )io-accu&ulation -An in%esti#ation on the eect o hea%$ &etals li.e /ercur$ an,
0a,&iu& on 1resh (ater ish Clarias batrachus carrie, out in the la)- 1ishes (ei#hin# 10 2 0-5 # an, 10 2 1
c& len#th (ere e*pose, to su)lethal concentrations 3,eri%e,+ )ase, on acute to*icit$ tests4 o &ercur$ 0-15+
0-05+ 0-05 an, 0-03 pp& an, o ca,&iu& chlori,e 0-66+ 0-30+ 0-17 an, 0-12 pp& or 30 ,a$s- 8urin# this perio,
ishes (ere e, (ith artiiciall$ prepare, oo, -9n the 10
th
+ 20
th
+ 30
th
,a$ ishes (ere ta.en out+ sacriice, an,
the tissues o #ill an, li%er (ere e*cise, out- "he total autops$ (as co&plete, in less than 4 &ins- "he result
sho(e, that the ,e#ree o ,istortion o the #ill+ li%er (as proportional to the e*posure perio, an, concentration
o the &etals (as oun, to )e ,ose an, ti&e ,epen,ent-

INTRODUCTION

/ore an, &ore o our ha)itats are )ein# ,eteriorate,
,a$ )$ ,a$ ,ue to increase, en%iron&ental pollution
)$ &eans o %arious anthropo#enic acti%ities- "he
in,ustrial eluents that contain to*ic su)stances li.e
hea%$ &etals+ pestici,es an, other che&icals are
,ischar#e, into the (ater )o,ies- An$ pollutant
(hich is ,ischar#e, into (ater (ill chan#e the
surace tension+ ther&al properties+ con,ucti%it$+
,ensit$ an, p: %alue alon# (ith )io,e#ra,a)le
3proteins+ ats+ pestici,es+ un#ici,es etc4+ an, non-
)io,e#ra,a)le pollutants- "he aci,ic an, al.aline
pollutants ,estro$ &ost o the in%erte)rates an,
&icroor#anis&s 3!ar)er an, Shar&a+ 15584- As a
result the a'uatic auna an, lora are a,%ersel$
aecte,+ (hich lea, to )ioaccu&ulation in a'uatic
or#anis&s an, )ioconcentration in hi#her %erte)rates
3Ac.er&an+ 20014- ;co-rien,l$ en%iron&ent is a
necessar$ con,ition or the (ell )ein# o hu&an
race- "he ,e#ree conta&ination in a'uatic
en%iron&ent is re'uentl$ assesse, )$ co&parin#
contain&ent concentration in associate, )iota-Since
)ioconcentration o co&poun,s ha%e )een
,eter&ine, in the en%iron&ent +it has )een o)ser%e,
that there are &an$ 'uantitati%e relationships
)et(een structure an, )iolo#ical acti%it$ o
che&icals esta)lishe, in a'uatic s$ste&- :o(e%er
the &ain sites o these hea%$ &etal upta.e an,
accu&ulation are the #ills an, #astrointestinal tracts-
Lea, is a co&&on hea%$ &etal oun, in the
en%iron&ent an, is ,eri%a)le ro& ur)an (aste
(aters+ in,ustrial ,ischar#es an, a#riculture runo-
Its inclusion in #asoline as anti-.noc. contri)ute to
its occurrence in the air +(hich is transporte, to the
strea&s an, ri%ers )e runos (here ish an, other
Research Article ISSN 2250-0480 Vol 3/Issue 2/Apr-Jun 2013


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Lie Science !io "echnolo#$
a'uatic or#anis& ta.e it up an, incorporate it in their
)o,$- "he ,eath o thousan,s o Japanese (ho ate
ish ro& /ina&ata !a$ in 1552+ the su,,en ,eath
o 3or4 4+000 people in Lon,on in 1560 ,ue to eatin#
o ish an, si&ilar e%ents in,icate the necessit$ or
ensurin# a clean en%iron&ent- As an in,icator o
en%iron&ent li%a)ilit$+ ish see&s to )e %er$
i&portant since it is aecte, )$ the li%in# ha)itat
3na&el$ (ater4+ (hich is to,a$ pollute, )$ eluents
ro& in,ustries+ pestici,es (ashe, out ro&
a#ricultural lan,s an, ,eter#ents ro& househol,
,rains etc- "his paper is ai&e, at ,eter&inin# the
#ills+ li%er+ o the Clarias batrachus to su) lethal
concentration o ca,&iu& an, &ercur$-

MATERIALS AND METHODS

:ealth$ li%in# speci&ens o C.batrachus (ei#hin#
10 2 0-5 #& an, 10 2 1 c& in len#th ha%e )een
)rou#ht ro& "a&ilna,u isher$+ <oon,i+
"hiru%allore 8ist- "he$ (ere )rou#ht to the
la)orator$ in (ell-aerate, containers+ to a%oi,
h$peracti%it$+ ph$sical in=uries an, stress to the ish-
"he ishes (ere screene, or an$ patholo#ical
s$&pto&s an, (ashe,(ith1> ?/n94 solution- "he
health$ speci&en (as then transerre, to #lass
a'uaria 350 * 25 * 25 c&4 containin# tap (ater- "he
ishes (ere accli&ati@e, to the la)orator$ con,itions
or 15 - 20 ,a$s prior to e*peri&entation- 1ishes
(ere e, (ith artiicial pellet ,ietA (ater (as
replace, (ith clean (ater (hene%er necessar$- "he
<h$sicoche&ical characteristics o the (ater (ere
anal$se, as per the proce,ure o 3A<:A+ 15554+
A&erican pu)lic health association- "he test ish (as
i,entiie, to the species le%el- Si@e an, (ei#ht (ere
ta.en the ishes (ere precon,itione, to unpollute,
tap (ater or 20 8a$s-

Preparation of Stock solution
/ercur$ an, 0a,&iu& 0hlori,e (ere o rea#ent
#ra,e- 0oncentrate, stoc. solutions (ere prepare,
)$ (ei#hin# correct a&ount o the salts an,
,issol%e, in (ater 31#/l4- Ali'uot %olu&e o
calculate, stoc. solution to $iel, the ,esire,
concentration (as a,,e, to the tan. to #i%e the e*act
re'uire, concentration o the hea%$ &etal-In the
present stu,$ 1/10+ 1/20+ an, 1/30 o the 56h L0
50

(ere selecte, as su) lethal concentration an, the
ishes (ere e*pose, to each concentration or a
perio, o 10+ 20 an, 30 ,a$s- A control )atch
correspon,in# to each test #roup (as si&ultaneousl$
&aintaine,- "he e*peri&ents (ere repeate, i%e
ti&es an, concentrations supplie, ,ail$ to &aintain a
constant to*ic &e,ia- 1ishes (ere ta.en out+ )lotte,
,r$ (ith sot a)sor)ent paper an, ,issecte, to
re&o%e li%er an, #ill tissue- "he or#an (as
preser%e, in la)ele, sa&ple )ottles containin#
or&al saline+ sectione, an, sli,e preparations (ere
&a,e or histolo#ical in%esti#ation un,er the
&icroscope-

RESULT

Histological Study
Effect of Mercury on Gill
"he histopatholo#ical stu,ies in,icate, that the #ill
o C.batrachus (ere treate, )$ su) lethal
concentrations o /ercur$- Ater 10 ,a$s 3<late 1A4+
&ini&u& chan#es (ere o)ser%e,- Not &uch chan#es
(ere o)ser%e, in <ri&ar$ la&ellae )ut the secon,ar$
la&ellae (ere use, to#ether to &ost o their len#th
(ith the ter&inal 1/3 portion ree 3<late 1!4- "he
inuse, secon,ar$ la&ellae (ere thinner co&pare,
to their controls- "he epithelial cells (ere seen in
)et(een the use, secon,ar$ la&ellae- In so&e case
secon,ar$ la&ellae appeare, to )e cur%e,-In 20 ,a$s
treate, ish+ pronounce, chan#es li.e ,istortion o
secon,ar$ la&ellae+ h$pertroph$ o #ill ila&ent an,
h$perplasia o epithelial surace+ ,etach&ent o
epitheliu& ro& un,erl$in# pillar s$ste&+ the usion
o secon,ar$ la&ellae resultin# is a re,uction in the
surace area o the epitheliu& (as notice, 3<late
2!4-In ish treate, upto 30 ,a$s+ the chan#es
o)ser%e, in the #ill o
C. Batrachus (ere se%ere erosion an, ,e#eneration
o #ill epitheliu& an, a##re#ation o )loo,
corpuscles- 1usion o the !oun,ar$ o the secon,ar$
la&ellae increase, (ith e*posure perio,s 3<late 3!4



Research Article ISSN 2250-0480 Vol 3/Issue 2/Apr-Jun 2013


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Lie Science !io "echnolo#$
(late 1
Histological lesion in Clarius batrachus (Cat fish) exposed to
Mercury and Cadmium Gill ! "# days





(late '
Histological lesion in Clarius batrachus (Cat fish) exposed to Mercury and Cadmium Gill ! $# days


Research Article ISSN 2250-0480 Vol 3/Issue 2/Apr-Jun 2013


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Lie Science !io "echnolo#$


(late $
Histological lesion in Clarius batrachus (Cat fish) exposed to Mercury and Cadmium Gill ! %# days





Effect of Mercury on &i'er
In Clarias batrachus treate, or 10+ 20+ 30 ,a$s+
lar#e area o li%er tissue sho(e, ,isrupte,
hepatoc$tes (ith loss o nor&al palisa,e
arran#e&ent- Ater 10 ,a$s o e*posure+
,e#eneration o )loo, %essels+ %acuoli@ation an,
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Lie Science !io "echnolo#$
h$pertroph$ (ere o)ser%e, 3<late 4A+ !4-"he
hepatoc$tes appeare, s(ollen ater treat&ent or
20,a$s 3<late 5A+ !4- "he li%er sho(e, acute
inla&&ation inclu,in# rupturin# o hepatoc$tes an,
increase, accu&ulation o p$.notic nuclei+ necrosis+
%acuoli@ation o tissue an, ,e#eneration o )loo,
%essels- At the en, o 30 ,a$s e*posure+ the li%er
tissue ha, lost their c$toplas&ic ,ensit$ an,
appeare, opa'ue in &an$ areas- 8e#eneration o
hepatoc$tes sho(in# ,istinct %acuoles+ necrosis (ith
sinusoi,al lesions (ere the &a*i&u& alterations
o)ser%e, 3<late 504-

(late %
Histological lesion in Clarius batrachus (Cat fish) exposed to Mercury and Cadmium &i'er ! "# (ays



A& ontrol


B& Mercury











Research Article ISSN 2250-0480 Vol 3/Issue 2/Apr-Jun 2013


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Lie Science !io "echnolo#$
(late -
Histological lesion in Clarius batrachus (Cat fish) exposed to Mercury and Cadmium &i'er ! $# (ays



A& ontrol


B& Mercury


& admium

Effect of Cadmium on Gill
In ish e*pose, to su) lethal concentration o
0a,&iu& %arious histopatholo#ical alterations are
o)ser%e, ater 10 ,a$s o e*posure 3<late 104- "he
secon,ar$ la&ellae are thin an, are use, to#ether-
"he shape (as altere,- ;rosion o h$pertroph$ o
&ucus cells+ epithelial cells+ ,e#eneration o pilaster
cells an, epi,er&al cells (ere notice,- "he usion o
secon,ar$ la&ellae &a$ ha%e )een ,ue to e*cessi%e
secretion o /ucous-In 20 ,a$s treate, ish+ chan#es
li.e ,istortion o epithelial cells+ h$pertroph$ o #ill
ila&ents+ h$perplasia o epithelial surace (as
e%i,ent- Re,uction in surace are o the epitheliu&
an, ,etach&ent o epitheliu& ro& pillar s$ste& (as
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Lie Science !io "echnolo#$
notice,- 3<late 204-At the en, o 30 ,a$s+ as the
,uration o treat&ent increase,+ &ucous cells
increase, alon# (ith ,istortion o #ill ila&ents an,
la&ellar re#ions- Se%ere e,e&a+ separation o
respirator$ epitheliu& an, %acuoli@ation alterations
(ere o)ser%e, 3<late 304-



& admium



& admium



&admium





Research Article ISSN 2250-0480 Vol 3/Issue 2/Apr-Jun 2013


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Lie Science !io "echnolo#$
Effect of Cadmium on &i'er
In ish at 10 ,a$s o treat&ent+ s(ollen hepatoc$tes
(ere notice,- 8e#eneration o hepatoc$tes+
%acuolation space or&ation (as e%i,ent+ throu#hout
the tissue 3<late 404- At 20 ,a$s o e*posure an,
hi#h concentration accu&ulation o )loo, %essels+
con#estion o hepatic tissue (ere also o)ser%e,
3<late 504- Ater 30 ,a$s o treat&ent the li%er (as
hi#hl$ ,a&a#e, su)capsular %acuoli@ation+ necrosis+
in,istinct cell )oun,aries in &an$ places an,
p$.notic nuclei (ere also o)ser%e,- As the ,uration
increase,+ se%ere ,e#ra,ation o the li%er cells or
hepatoc$tes an, h$pertroph$ o hepatic nuclei an,
clu&pin# (as e%i,ent in &an$ places 3<late 604


& admium

(late "
Histological lesion in Clarius batrachus (Cat fish) exposed to Mercury and Cadmium &i'er ! %# (ays



A& ontrol
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B& Mercury


& admium

DISCUSSION

"he histolo#ical chan#es on ish is a note(orth$ an,
pro&isin# iel, to un,erstan, the e*tent to (hich
chan#es in the structural or#ani@ation are occurrin#
in the or#ans ,ue to en%iron&ental pollution- Sin#h
et al.315504 stu,ie, en%iron&ental pollution an, its
eects on a'uatic ani&als-:ea%$ &etal to*icant
lea,s to &an$ patholo#ical chan#es in ,ierent
tissues o ish an, has )een reporte, or Labeo rohita
e*pose, to &ercurichlori,e an, Chana punctatus
e*pose, to phen$l &ercuri acetase 3?aruppasa&$+
20004- Sri%asta%a et al- 315824 ha%e o)ser%e, the
hisotopatholo#ical chan#es an, accu&ulation
potential in the ish tissues un,er chro&iu& stress-
0a,&iu& e*posure in,uces the appearance o
#ranular ,eposits in the li%er+ atroph$ o the
pro*i&al renal tu)ules+ an, increases in chlori,e cell
turno%er at the #ills 3<ratap an, (en,elaar !on#a+
15534-Bill h$perplassia+ necrosis o intestinal
&ucosa+ at iniltration o li%er parench$&a cells+
,estruction tu)us (ere o)ser%e, ,ue to a&&onia
poisonin# in the intestinal tract+ .i,ne$ an, #ills o
=u%enils Sparus auratus 3Ca.i et al-, 15874-Bill is an
i&portant tissue )ecause o its ,irect contact (ith
(ater an, an$ eect o a#enc$ has to #o throu#h it to
co&e into the ish )o,$ 3Jana an, !an,opa,h$a$a+
15874-It has )een su##este, that (ater)orne a#encies
,a&a#e ish #ills+ presu&a)l$ )$ causin# )rea.,o(n
)$ the #as-e*chan#e &echanis&s (ith the
conse'uent tissue h$po*ic con,itions 3Natara=an+
1585a4-"he &icrophoto#raphs o #ill o C. batrachus
ha%e )een o)ser%e,- In control ish+ the structure o
#ills is si&ilar to that o other resh (ater cat ish as
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Lie Science !io "echnolo#$
,escri)e, )$ Laurent+315854 (here as the #ills o the
e*peri&ental ish e*pose, to ca,&iu& an, &ercur$
310+ 20 an, 30 ,a$s4 sho( acti%e secretion o
&ucous- /ore speciicall$+ in the &ercur$ treate,
#ill the epithelial la$er (as ,isrupte,- In e( re#ions+
,isinte#ration an, usion o pri&ar$ la&ella (ere
o)ser%e,- In ca,&iu& treate, #ill+ &ar.e,
h$perplasia o the )ranchialarch+ pilaster cell
%acouli@aton an, con#estion o )loo, %essels (ere
(ell &ar.e,- Si&ilar results ha%e also )een o)ser%e,
)$ S.i,&ore+315724 in rain)o( trout (hen e*pose,
to @inc sulphate- :istolo#ical e*peri&entations o the
#ills o rain)o( trout )$ Llo$,+ 315654 (hen
e*pose, to @inc+ lea, an, copper sho(e, separation
o the epitheliu& ro& the #ill la&ellae an, cells
slou#he, o into the spaces )et(een the ila&ents-
"hese chan#es are in a#ree&ent (ith the present
o)ser%ation (hen the ish is into*icate, (ith the
hea%$ &etals &ercur$ an, ca,&iu&- "he results are
also in arran#e&ent (ith the (or.s o ?apila&ano=
an, Ra#otha&an+ 315554 (ho ha%e reporte, or
Boleopthalmus dumieric e*pose, to su)lethal
concentration o ca,&iu&-In the present stu,$
%acuoli@ation+ space or&ation an, resultin#
hae&&orha#e+ h$pertroph$ o hepatoc$tes an,
clu&pin# (ere o)ser%e, in &ercur$ treate, tissues
an, e*tensi%e necrosis+ p$.notic nuclei in ca,&iu&
treate, li%er-?a)ir an, !e#u& 315784 reporte,
c$toplas&ic ,e#eneration+ p$.notic nuclei in li%er
tissues+ %acoulation in hepatic cells an, rupture o
)loo, %esselsA ,e#enerate hepatic cells an, necrotic
nucleic (hen Heteropneustes fossils (as e*pose, to
C. punctatus to a su)lethal concentration o
en,ocrine an, o)ser%e, h$pertroph$ o hepatic cells
an, li%er cor, ,is rra$+ %acoulation o c$toplas& an,
necrosis+ ruputre o hepatic cell &e&)rance an,
necrotic centrol)ular area-"hese t$pes o histolo#ical
alterations (ere also notice, in the li%e tissue o
Labeo rohita e*pose, to tanner$ eluent 3Rana an,
Su,hir+ 15554- "he li%er tissues o L. rohita sho(e,
histolo#ical chan#es ,ue to alato*in #i%en to ish
intraperitoneall$ 3Sahoo et al-, 20014-0onclusi%el$+
&etals are store, in ,ierent sites in ani&als
,epen,in# on the &etals an, on the ani&al species-
"o chec. the continual intro,uction o these &etals
in to the oo, chain+ a &ore cautious application o
insectici,es an, pestici,es shoul, )e e&plo$e, an,
eluents ro& in,ustries &ust )e treate, )eore
,isposal-

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2- A<:A 315554- Stan,ar, /etho,s or the
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3- !ar)er 8+ Shr&a /S 315584- ;*peri&entall$
In,uce, !ioaccu&ulation an, ;li&ination o
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Research Article ISSN 2250-0480 Vol 3/Issue 2/Apr-Jun 2013


L - 21
Lie Science !io "echnolo#$
:ealth$ an, 8isease, 1ish 3;,- S- Eoo, /arcel
an, 8e..ar4- )nc. *e+ ,or&., 65-120-
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Seminar, (.S. ept., Health $ducation and
Welfare., pp- 181-
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0oncentration o /etas$sto* on Selecti%e
9*i,ati%e ;n@$&es+ "issue Respiration an,
:ae&atolo#$ o the 1resh(ater Air-!reathin#
1ish+ Channa Striatus 3!lee.er4- Pest. Biochem.
Ph.siol., 21D154-158-
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0eils+ 0hlori,e 0ells+ an, NaF / ?F Atpase
Acti%it$ in the Bills o the 1resh(ater "eleost
/reochromis 0ossombicus at Nor&al an, :i#h
0alciu& Le%els in the A&)ient Eater- Aquat.
"o1icol., 26D 133-150-
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Alato*in B
3
to Rohu+ Lobeo !ohita 3:a&ilton4-
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0a,&iu& in 0ertain 1resh(ater 1ishes o Ri%er
Su)arnare.ha- J. 'resh Wat. Biol., 2D 185-153-
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Sulphate on the Bills o Rain)o( "rout- Wat
!es., 6D 217-230-
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315824- 0han#es in Bill o Punctius Sophore
Gn,er 0hro&iu& Stress- J. Biol. !es., 2D 85-
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Bull )nst /ceonogr. 'ish Cairo-, 13 314D 43-57-