teen Manet hicasy Fach A Scanning and Transmission Electron Microscopic ‘Study on the Effect of Environmental Acid Stress on a Hill Stream Fish (Brachydanio Rerio) Gill Shiv K. Prasad’, Sudip Dey'-, and Q. P. Singh® " Sutisxet rail rune Fc, Noch Est Hl Umer, og 88022. Mop, de "eprint of anata Stes Hor ave Mf Urs, Sir 802 Ngai Fe ‘Scamming and tersrisson elocton microscopes cn ha gf fa hi cream te, rachydanie ‘one, ihabting same streams of Mochalaya a Norh East lien Slate) wi scl pt read ‘Sunber of abrenmaftes. The ethos mere eeusurl sergrmoties nace le of eigrent ot inary end secondary it meta, csorion shirkage, deposi of mci one arate Beeiee rece, fusion of i lamola at placa, and dovelooment of Sagan He pen on fhe ‘race of fe pf lama wete en Tae fe socal abremaitee te lee four Com. tour el the fp of secondary Git mela, eegkationsollamedar membrane al he tp std ern ‘ne micas part ol he rata ta scant exert. Al these uta ruck abesraiin in ne On response to endionnanen acdsee nthe nti Rabi ot Sacha enDie ecu With eel of rlsvant Serve. Keywords: Ackkc Vise, Envrcrmeria! Stress, Hil Seam, Scaning Cheon Meran2oy “Yanemiesn Secon Mcrosopy Fh, GL 1. INTRODUCTION “The survival and grou ofS and other agate espa fens greatly depen! on number of contin och 45 altude, exnperaure sad Rmaoloical factor. The lndueace of haba stuctse cm tar compention snd ‘opalson dynamics of fs s well now” Te is amply highligh in exsing Teramre dha hi- tat degradation and advese change in water qiality of ‘he aque covioariont aw the most import factors Tohuencing the Bch rowehsepedacion aad mxvial has does seporcd at among erent ate fc- vor, pit pays an omporan ole in growth ariel and ‘eoduclam eh? The pH vale exposes he ttety of acidic or tasks character of mater xpos of 81 ‘oes of pl can be sess rea Tae aie pi of ‘ale is lows Wo cause dsrton in the fh population dynamics by high ae of moray daring carly a well at lee siges tm te hfe hsty-™ Ihas bee reget that ‘ed stess was seaponibe for near extinction of ss [opulaon in Ons, Canada“ Low pl hus been pod ‘oeame sacral damage al muscle, ia and pore ‘ofa sex fish Danfoauapinanis oleted from he ar © wom compte ta bette ‘wal Raitt with acidic pH. The stot abaoraly (Ed not oocar when the sh was cece in aguas where tral pH wan mantis ‘Tac retin beeen acidic pH of water and sein in growth and rprdcton in some fch fas 0 ema ‘eonfined Gonz experimental sis” let of environmental ci ses ia bicaccmation of cerain heavy metak and increased aserpin of pt ‘ies in tse of some fish and ether aque organi "as bce por hy some sntors*° Distasbances in ekmental homeosase snd increased izwrption of coma pesices ia regent 1 secic PH of water in some fs have also too report ‘esta and Kiama (1095)! showed he adver eet of low pH exposure of ada ealnonids on gametogenee amd enbre developaeat was et) (1990) edie he ‘fect of sulphuric ac cues om oxnoreguitcy ability of Tapanese Char (Selous Imcomeert), Elevation of Dlsma somatnisin concentration tor been sepried So ‘ainbow out (Onchoryctur mpi) ding sacs © ‘Sevae suppession of spauning. im hime salmon (Onchorlyechr ner) de ace ot water his bose ‘epaed” Adi pH bat been fouad to have dfferat tolerance in dierent eh spaces Bosdos this, coral seaniipnstnom TOW HouvaSaHRESEARCH ARTICLE ‘A Soaming md Tessin Blcooa Msceopi tayo Est f avin Aid Sess dierences in ci olor have bon epried im Maisx onyalepe by some sxthors Dey ot a (200) reported thats te eesamcatl tolerance of sone vestbmis Ins been tpt Re. [2 I ih, gil aes exposed ote xvaoumest and ‘he hey maybe wad acest of water gel Fh 2 ply vial rs sine they ar the maint of asons cxchanee™ "Tam tn eviden hat St il amb wed mx ede fac soies on envroamental part of acid sre” ‘Alboogh ecuogies, pigselogial aad” Sochenical idles cn acd snes in Sch Deve teen cane out by fone ston," les stu stdie ae Inking i is part Keeping fis x vw, a Scaming and Tesnamistoe heen Micrscopie sty on the gl of Brechydano ‘ono with referee to emizoamentl aid atest iat ‘mural Asta ban boos le up. Seuasing lec ‘misosopy Recane of high resolving power an age depth of lds Healy to rescat deal sarfoce micro ‘srctal sbaormaies inching aigament ofthe gil (urponcns. wie Taunton clesnon micoopy ae tors vey high esclton and cepiicion expec to bing out ie sacral efits, i amy, in te i, fend by eaviroament a sess 2. MATERIALS AND METHODS 2. Fish Materials Fi species ected forthe stay nos Brachydans vr. “The collection was mace fom “Ung” sean aed “Wak-Unmah ste” sted In and sound Sten, the capt iy of Meghataya, 2 Nerh East Indian Ste. Moutly colecsont ver made fe deen #23. ‘Ss (quing Sommer, otis and Wine) aug he ye amr 08 22. Memsorement of PE Water pf he seen was measured every Week with Iielp of pA Moc (U pi System 361, Sperone) ing the stay peo ‘23, Esperimental Set-Up “The joven fh collected from the sata habitat with antic pl were dod ito two 20093. ‘One of the groups was ror in aquariom wit water foo the matali, amd side pH as ened ee ‘he mal hai the pH of wae war changed allan vale by te dion of NaOH Bo he grows (joc ice were ee i the aan ty eset ey 224 Un Stracsral Seales “The Sich collcte fom th natal abit a well Soak ‘ao expenmensl aguas were aneribcied by wag Te ‘ine methme sulphonic and gills wor icin for ‘loasaectral studios Ten gil samples each fom fh feowe in steal tat and frecn eapeiatl growpe ‘wer seed for als sinctia smaysr dering cach so Si of the sly period 0 confirm npc of the 2ukd. Scanning Electron Microscopy ‘The gl samples exssed tom the Set were eat Sato small pies (222 mm) and wee fine in 29% ga. ‘ecldetyde prepared in OL M Sodium cacodylate butler (GH 72), cehydte though increasing concctraon of ‘secloge and were died using TMS following the meod (of Dey et al (0989).° The dry samples were secured bore Faomly mo bess sie (10 en dices 30 igh) ith dosble shy tape. The samples were then contd ‘ith goltpaladinm in JRC 1100 (EOL) lou Speer Cote and were cbned in the sexondary eicron eit Sion mode of + SM 6360 (IEDL) suming ceewon mio ‘cope stan aceloming vise of 20 KN. WD selector ‘yas ued io mate the working istanee (WD) 3 om 242, Trenemicon Elcom Merscopy ‘The samples wore cut into small pices of approx: smacly | m1 si nse and wore fd io mie Kamovaiy’s fiatve™ heving he ompostion of 250 ai of 02M Soc cuca bf 20 of Pra fermae- Ine diswoled i ita 60°C iaging te vote 1 480 ‘by do sled water. To ds, 2 lf 25% gare Iiyde and 125 g of airs air sre wor de Aer foer hous is tbe above pimary ashe, the sy Des were wasbed throughly i 0.1 M Sockameacody- Ine batle Pos: futon wer cad out i 1% Osman ‘eerie prepa in he same lle for oe hor at 4, ‘Spocinent wee dhyated in ascading pales of ace- ton (20%, 50%, 70%, BOM, PE, 95% amd dy ace ‘one) wat two changes of 15 annex each The spl Ae ee. Roe 61-8 2044sree thn led ff the aso by prope onde for TaSetion was cad ont grade in rent gro porous of propylene onde with he ented eon Digui ein Araki CY 212—10 mi; DDSA desceyh ‘occnicaniyide)—10 ma: OMP-30 Diana ro eaedaing medion ig Wem cape ‘The embeded Nos wore kept a 50°C ian emhed 'e 60°Cand thecal tet were hop fr 18 boars ' compl plyercaton, ‘Ueahn sections (600-800 A) were ca in aw RMC Ute Mierotome, MEX with 2 dammed tafe The seciom we cllied on copper grids and ned wah 5% aleshotc soliton of Ural acct fn 10 miwes stor tperatire nthe dk, lle Led ie “he shaved seine were exaied ina JEOL JEM 100 (CXL ramen Electon Micrscoge i sm acters volage of 80 KY 3. RESULTS 1 Uitre Strctaral Stes LI. Seanaing Flctrom Micrscopy Tih grown at leaine pH shows! nomad Keates of {Bll wl pop agnmnt and wel ined prey ad Secvetay mela 1) 1m the alle of fo colecte fom its earl hata wth ace pl he aigenene of he pry and soo Ainy gil mets wae fond to be fost (ie 2, Bess ti, tee mere dsr of sescmday gil Enea ‘aces ad depostion of nce atthe sare of the 21 lemel parcataty al the ip (Fe. 35). act ot, shinoge and distor ere cident at te ip ‘ot cme gill lanella (Fg 9) Hyperpiia, characaied Ade Meo: os 6, 1-8, 2011 1 Sconing nd sms acta Mie Sly te Fe of orm Sess Te 2 Sattiy coon memgagh evi nw of gon of by ceil policeon in interarm mefon leo int (7). ‘son oF lama at ples, brskage at the base of primary il le (Pip) a ep detachment 3 SOUY HoUWaSayRESEARCH ARTICLE Of inary il lamella, were obered at some ples eo) esos eset, breakage, Is f alignment cc, the evelopment of some fnge print lke patens on be ce and De lame wee abo 1). Sina acral sbmore shea sh agent tac pi ch pou a alae pid ot crn crak il ice rte of the gill 842. Transmission Heron Micascomy “The owns coaaur fhe secondary il lanes was ay stared atthe op (es. (x9) compare tec tol (Pgs. 126.8). The menbrase of he il aed a ‘he tp wr ound be evap paces Fart i ‘le prin of the fame wat fone Wo be Gnd to ‘tenicent exten ig. 11). “The side shaped led cells which cnt somal feamge in contol wae fome 0 be str tthe ah {row af cle pH fi natal bit ig.) ‘The tood coll were found to be clumped apes (fg. 11a) aed regularities of te cesta an periph pats mer lost completely (Fig. 13) The mle gut of tome of the sil lames showed outa dies 0 te smembnme Fg. 1RESEARCH ARTICLE. ico snctvals>eonaies in be gil in tespomse $id oe er eat y 7-0 oe fh a esd wee sca terrain ave repletion 0 Inent nar the water sfx and more frequent opera ‘oven’ a stated conol eam stan 10% Imaal was herve in borin eae sh nae elu However sir wesprakey tess mera iy we Oe born rd poptn 4. DISCUSSION The ctnrvaon male bn th peen dy sugges tat simormaiics extibvied by gil he eh lc! fm incnatnl habit we due © he ade pH of eet. Tas fs tupac ty te fact that when rei Ies neerene to mata in again with wate ro the at Sra ha, ote Ht mat mae alba, cal be stuctarssboealites st shown by fh fm na Tablat were cashed by jini fh tated ay in wer hic catalan seal ae Bs nochange Dirt id of raeee expend by fh citer thongh chercally indeed chee oc dgh ‘ames ses caney = eomber of abermalie In (mopkgy and ine strc of fabon Th or preset say he ssoning lc mice seectim of eof alloca af te pny an = ‘airy gil acl intestate an cease fo {ie pocton aioe tothe Bal acl Th ess in ‘acened diffi dtc whch town to cae po, ‘fem in gas nchage. Tis era iin dee is dr tn cane in ole of he ton ae cps ae secondary Tamer pcm” Father, he Tos of aligenent was rgd Yo ‘ove sedation in pce feween mghioang comay Tabet which Kel to ave severe aver alto ‘change: Citing of ect) ea ng fen paw as cbc ne cre sy es te So “hese abeormatses ar hly to ens isis ia sos change despre. & mls typeof ce ‘ation vas made in hyve es of he eh rah Sone re (Harton Boeken) sncnced by Cann ove This cigars ht mesos and i taal darnaiies ia Fh pill way be came che by tne chemicals cr by cmos test deo ‘oor water gealy (ide of wax Oe sem coe) ‘Absormal cra ease gills revealed fom seaming oeeuoe miemsenpe Mike in eer ieee Shon fey cae diese espa mechan and also ote anton of the gil ach a tn el on a his comfy 4 1 He sd tat eer med sss erwin ius im cums rpc Te abe 1s farts the anomiston lee rkosogic st sahons a coveued. entra rin onl ‘Genin comiurof he up ingle ae oe of Cece: so membrane, coig clr mcrae Jesdng © prvi ott The beakage and disor ton of il artis memtesme ad os of ee ees Img aera ceran caer wars Defamation of spines od cas in seoadry lane aa hemsigil groblers- Silly sak ‘en disenson inthe mide panto of toe of all Inrela nce cell wilerion which way lea pein Fare, te ean alo sncease the fsa ice tetaer gh i linc ‘lear defrost an Toe of elle fete in i Taela gues nt expose of th eh ee pt as Ao me fee. 89-8, 2097 A Somming Wasisn Eimn Miso Sno he tf asa Ad Ss cause severe eller shmcrmaiies, Hang bo neti of te tito noe ha otal esi mela ae averse ameied tn he gil ofan ind Fah evan fara alBoeph i ifenlty Neves sca normals i be gl ewer Inf mele can xe ‘aici phyla sea Se faking ey Steak neue, ‘Nbcomal sical Cues exit by aochondric Inching dorm of shape, dsetion of ot and at Imembrane ot inlews dlbealy se entty fena Eating o prob in eapcry fenton Mecin etal (UI) sagged a pg ‘el eunges i, partly the eich se chusewled bye fe of fon the aon Of me seine eeepton of sand okie inte _pmcows exchange Mallat (3989)° rosewes hate fhm dcuchnest i » comma pienonenon in git of Jn apenewene sere water quality. Now (1092) found hay ie fspiaoy epi dtsehment rs ‘in the nts in inion dates, ecting the gaa Kavall (19) jot at spratory pitta <€xachment it ils dae cnivommenal eenanieton Sock a5 acid atm some Bethe pep os Sstnegoat tte oes th edit detachment fSuggeting tthe former was comer of ater Te gpa se chert in ously chrcsied hy ‘ir clike poker net ae gion of ‘toy Imeardeceving these sea and mang scons change tre iia Mazer (18) seer tha tee pofraied cee stemmed fom the tur, ‘ofthe lament nth intramelr pae and acted a ar "er othe lain of oe ttn ote th hd [tong aa epost wdkam aes wes ualty wa oon ond tect of sane ues thle sistas te blood Bom tos hare the vere (Mfc of deoasng te aygennion fae, setae ‘hurler ip aceon onan ‘he poseace of mires ithe sch and benchi ‘amet of rahydann veri eae em theca ‘eatin ae karen oe clad fo the acon anchor Im oer to prec the eit inate ot Sod agents tn spnson Hower it hs ber see ‘ha he presace of sch sce the eel con ‘nce the Bites of Moura hare nich Gi tubs the pascons crane The eae need he Dit steam gown neat bt wih acdc pl dvelps the mien ides opoterthe epiil nt {he atnrpon of te vatatnce Sieh boy Hen ern the pes een the ae ithe thik ‘looarvaer maser and Kinde the pcos eich Ohdbone fle of be il Teaom at soared he foment sidy ae epeced ty inde Jattbues te ‘aTOWMY HouVvasauRESEARCH ARTICLE Ihe Helin epteliin eligi an increased Nowe ow imide te lamellae wick eam caer len oe ‘murs channel, Hoed congestion or even anes tis o be reed im wie content ht cea pill Jamel ae he ee of nsentseelboie acy and Ince the abel es feats i the gill mel ae Ince of some metabolic distances ing “The distances im the gill etme die to ade PHL of wats appar lo be ne fo cata shsormalites In imc meen ad ekctoenomisd! feds ass 19 whieh ca be everzome ony by cringe Fak nde sutra cr aa pt of mae 5. CONCLUSIONS 1a lear at emironmestl a esc case nam ber of abaormaites im al of the fish Prcdain ver Jeatog to Hs mae fnctoning. Sic il lays ay ‘mporint res meeps lee of ions waste, iba balance ce or prow Seaning 2a Trane Son sleccse mieapie dseraen em pil of he fh Fespom ose pl matali apps tobe Sienrean irc Te present say sages da seanng a ‘nanaision eco obeoscopy ean ply 4 vey por Saat rie sing the extn of ue shoo’ a ‘eh de to earronmentl ses al can serve the sep for free sac robin othe ota “The pesca sera us stances in Fah Used ip aie of water sapped by Cer ivesugaions volving deren gprs Ii Therefore ausee ted that clon micrmeestades ander aid sess «wre appa ace wil ‘rate to ald soe import infant be elt eater sis wong iret eprceces References and Notes RS cre kn ing 8 + Eien ng 1 an BT ie Siete a es {tannin apse. Po Ths Ca Uno paren SA seit Ean, Wasi lc 197 81 1B fegat age eo ih Souk 6 9. Spy ramen 1 aa, 1B Satpbo. SW Rambo MmacagesdF ha, Soar oon % Sd Rn aa Sat 3 1S Ea tm et ean 9 TCD eT Yen at? ot om 1 Sabp by knobs whan Ue cavemen rae 72 OO, 16 Chapt a or Dy Wien Gola iO 8 7, en tr tok Bot Tv 5,0 a 1 he Arm cay ty 2 ML Menon aR 3 Bichon, Po 98 0) 2% Sp Do 5. Da Ba aya. Dy Mere 2 EE ema 1m tk 1,208 9 2 OME See HL a 9 6 Mee Tia Ce 1 pa 2 (hanno Cham Ase The teed Fecal Pea Cth ra cm 12 Mier Con J 2a G54) ‘ocean 8 0 a, Pyne caey WS or dD I Rade, en Nos, {any si RUE ha at Hee 1G Po 6 eee are Received: 30 July 201), Accept: 20 September 2011 4 Ae arose. Hos. 6, 1-8. 2018