G.2064Page1GlobalResearchjournalofNaturalScience&Technology(GRJNST)Volume:04-Issue2(2026),2064ISSNP:2790-7643ISSNE:2790-7651www.grjnst.nethttps://doi.org/10.53762/grjnst.04.02.15Comparativeanalysisofsite-specificIntestinalHistologicalvariationsinAsianSeabass(Latescalcarifer)andSeabream(Nemipterusjaponicus)alongtheBalochistancoast,PakistanReceived:23December2025.Accepted:26February2026.Published:17April2026NaghmasabzalDepartmentofmarinesciences,facultyofmarinesciencesLasbelauniversityofAgriculture,waterandmarinesciencesLUAWMS,Uthal10250,Pakistannaghmasabzal@gmail.comEmail:WaliullahMasroorDepartmentofmarinesciences,facultyofmarinesciencesLasbelauniversityofAgriculture,waterandmarinesciencesLUAWMS,Uthal10250,PakistanCorrespondingauthordrwaliullah.marine@luawms.edu.pkNisarAhmedDepartmentofVeterinary,AnatomyandHistologyFacultyofVeterinaryandAnimalScienceLasbelauniversityofAgriculture,waterandmarinesciencesLUAWMS,Uthal10250,Pakistandrnisarujjan2009@gmail.comKiranMustafaDepartmentofmarinesciences,facultyofmarinesciencesLasbelauniversityofAgriculture,waterandmarinesciencesLUAWMS,Uthal10250,PakistanCorrespondingauthorkiranmustafa255@gmail.comArshadhussainDepartmentofmarinesciences,facultyofmarinesciences,LasbelauniversityofAgriculture,waterandmarinesciencesLUAWMS,Uthal10250,Pakistanarshadhussainmb@gmail.comEmail:KhushbakhtDepartmentofmarinesciences,facultyofmarinesciencesLasbelauniversityofAgriculture,waterandmarinesciencesLUAWMS,Uthal10250,PakistanKhushibaloch333@gmail.comSadiqRahimDepartmentofmarinesciences,facultyofmarinesciencesLasbelauniversityofAgriculture,waterandmarinesciencesGRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Page2LUAWMS,Uthal10250,Pakistanrahimsadiq20@yahoo.comAbstract:Coastalmarineecosystemsareincreasinglyexposedtoanthropogenicpressuresthatcanimpairfishhealthandcompromiseecosystemintegrity.Theintestinaltractoffishisparticularlysensitivetoenvironmentalstressormakingintestinalhistologyavaluableindicatorofhabitatqualityandenvironmentalimpacts.Thepresentstudyexaminedsite-specificvariationsintheintestinalhistologyofAsianSeabass(Latescalcarifer)andSeabream(Nemipterusjaponicus)collectedfromDam,Gaddani,andKundMaliralongtheBalochistancoast,Pakistan.Intestinaltissueswereprocessedusingstandardhistologicaltechniques,andquantitativemeasurementsofvillilengthandmucosalthicknesswereusedtoassessstructuralalterationsamongsites.Theresultsrevealedsignificantreductionsinvillilength(34.79±7.29µm,p<0.0001)andmucosalthickness(8.33±1.88µm,p<0.05)inL.calcariferfromDamcomparedtospeciesfromGaddaniandKundMalir,wherenosignificantdifferenceswereobserved.Similarly,N.japonicuscollectedfromDamshowedsignificantlyreducedvillilength(34.79±7.29µm,p<0.001)andmucosalthickness(7.11±1.67µm,p<0.05),whileindividualsfromGaddaniandKundMalirremainednormal.ThesefindingsindicatethatDamcoastalareaissubjectedtogreaterenvironmentalstress,highlightingintestinalhistopathologyasareliablebiomarkerforassessingcoastalecosystemhealth.Keywords:BalochistanCoast,HistologicalAnalysis,IntestinalTissues,Seabass,Seabream1.INTRODUCTIONThesensitivityoffishtoenvironmentalchangesandwaterqualitypositionsthemasimportantbioindicatorsforassessingthehealthofaquaticecosystems.Variationsinfishphysiologyandtissuestructureoftenreflectexposuretoenvironmentalstressorsandcanprovideearlywarningstoecosystemdegradation.Amongcommerciallysignificantspecies,Asianseabass(Latescalcarifer)holdsconsiderableecologicalandeconomicimportanceduetoitswidedistribution,highmarketvalue,andadaptabilitytodiverseenvironments(Shaw&Handy,2011,Vijetal.,2020;).TheBalochistancoast,extendingapproximately770kilometersalongtheArabianSea,representsoneofPakistan’smostcommerciallyandecologicallyvitalcoastalregions,supportingrichbiodiversityandproductivefisheries(Qureshietal.,2010).However,increasinganthropogenicactivitiessuchasindustrialdischarge,uncontrolledfishing,oilspills,andcoastaldevelopmentthreatenthestabilityofthisfragileecosystem(Herrera-Garcíaetal.,2021).Theseenvironmentalpressurescandisruptfishphysiology,altertissuestructure,andposepotentialriskstobothecologicalbalanceandhumanconsumption(Kalantzietal.,2016).Thefishintestineisavitalorganfordigestion,nutrientabsorption,andimmunologicaldefense,andisamongthefirstorganstoresponsetocontaminantspresentinfoodandwater(Salinas&Parra-Medina,2019).GRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Page3Histopathologicalchangesintheintestine,suchasgobletcellhyperplasia,villousatrophy,andinflammatoryinfiltration,serveascriticalindicatorsofsub-lethalstressinducedbypollutants,microplastics,andotherenvironmentalstressors(Walford&Lam,1993).Forseabass,thesealterationsnotonlycompromisehealthandgrowthbutalsoreduceresilienceagainstdiseases,therebythreateningtheirecologicalroleandaquaculturepotential(Shaw&Handy,2011).NemipterusjaponicusisabenthicanddemersalfishthatisfoundinthewesternPacificandIndianoceans.ThespeciesisfrequentlyfoundinPakistaniwateratdepthsof5to80meters,typicallyinschools,alongsandyandmuddycoastalareas.ItisoneofPakistan'smostprevalentandeconomicallysignificantfishspecies,makingasubstantialcontributiontothetrawlfisheriesalongtheshoresofSindhandBalochistan(Kalhoroetal.,2014).Asianseabass(Latescalcarifer),alsoknownasbarramundi,isawidelydistributedandeconomicallysignificantIndo-WestPacificteleostfishinhabitingcoastal,estuarine,andfreshwaterenvironments,includingthewatersofBalochistan,Pakistan(Shahaetal.,2020).Itishighlyvaluedforitsrapidgrowth,high-qualityflesh,andabilitytotolerateabroadrangeofsalinity,makingitimportantforbothaquacultureandwildfisheries(Pattarapanyawongetal.,2021;Mehaketal.,2025).Despiteitsadaptability,seabassissusceptibletoenvironmentalstressorssuchaspollution,habitatdegradation,andsalinityfluctuations(Mauluetal.,2021).Seabream(Nemipterusjaponicus)isalucrativecommercialmarinespeciesthatiswidelyspreadthroughouttheBalochistancoast,supportinglocalfisheriesandcontributingsignificantlytotheregionaleconomy.Seabream,abenthicspeciesthatdwellsinsandyandmuddysettings,isveryvulnerabletooscillationsinwaterqualityandsediment-boundcontaminants,makingitasuitablebioindicatorofenvironmentalhealth(Khanetal.,2017).Histologicalexaminationsofitsprimaryorgans,particularlytheintestine,provideinformationregardingtheeffectsofanthropogenicstressonfishphysiology.Modulationsinvilliform,mucosalthickness,mayreflectsite-specificstressconditions.MaterialsandMethods2.StudyAreaTheresearchwasconductedalongtheBalochistancoastofPakistan,whichstretchesabout770kmalongtheArabianSeaandsupportsdiversemarinehabitatsandfisheries.ThreesamplingsiteswereselectedasDam,Gaddani,andKundMalir.Damischaracterizedbyartisanalfishingactivitywithrelativelylowpollution;Gaddaniisheavilyinfluencedbyshipbreakingandindustrialdischarges;whileKundMalir,locatedwithinHingolNationalPark,isconsideredcomparativelypristine.GRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Page4Figure1:(a)MapofPakistanshowingitstopographicalsetting,(b)AccessibilitymapofBalochistanhighlighttheregionallinkage(c)DetailedmapofthestudyareawithsamplingpointsFivetoeightsamplesofAsianseabass(Latescalcarifer)andSeabream(Nemipterusjaponicus)werecollectedfromeachsite,withweightsrangingfrom110to240gramesbetweenSeptember2024andJanuary2025withthehelpoflocalfishermenusingtraditionalgears.Aftercapture,fishweretransportediniceboxestotheHistologyLaboratory,DepartmentofVeterinaryMedicine,LasbelaUniversityofAgriculture,WaterandMarineSciences(LUAWMS).Standardmorphometricdata,includingtotallengthandweight,wererecordedpriortodissection.Intestinaltissueswereexcisedforhistologicalanalysis.2.1HistologicalProcedureIntestinesampleswerepreservedin10%bufferedformalinfor2448h,dehydratedinascendinggradesofethanol,clearedinxylene,andembeddedinparaffinwax.Thinsections(5–7µm)werepreparedusingarotarymicrotomeandmountedonglassslides.Routinehematoxylinandeosin(H&E)stainingwasperformedfollowingstandardprotocols.2.2MicroscopicandMorphometricExaminationPreparedslideswereexaminedunderalightmicroscope(LeicaDiaplan)at100magnifications.Histologicalfeaturessuchasvillousheightandmucosalthicknesschangeswereassessed.DigitalphotomicrographswereGRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064captured,andmorphometricparametersweremeasuredusingImageJsoftware(v1.51f).2.3StatisticalAnalysisDatawasanalyzedusingGraphPadPrism(version6).NormalityandhomogeneityofvarianceweretestedwithBartlett’stest.One-wayANOVAfollowedbyTukey’sposthoctestwasappliedtodetectsignificantdifferencesamongsites.Fornon-parametricdata,theKruskalWalli’stestfollowedbyDunn’smultiplecomparisontestwasused.Resultsarepresentedasmean±standarddeviation(SD),andstatisticalsignificancewassetatp<0.05.Page5RESULTANDDISCUSSIONRESULTSThecurrentstudydescribestheresultsofahistologicalanalysisoftheintestinesoftwofishspecies,LatecalcariferandNemipterus.japonicus,obtainedfromthreecoastalsitesinBalochistanDam,Gaddani,andKundMalir(Fig.1).TheDamandGadaniiscomparedwithKundMalirandisshownby*(Kruskal-Walli’stestfollowedbyaDunn’smultiplecomparisontest,p<0.05,N=3-5,21-25perindividualmeasurement.Dataareexpressedasmean±SD.4.1HistologicalAnalysisofIntestinalTissuesUsingHEStainingHematoxylinandeosin(HE)stainingwasappliedtotheintestinaltissuesofcommerciallyimportantfishspecies,LatecalcariferandNemipterus.japonicus,collectedfromdifferentsitesalongtheBalochistancoast.Thehistologicalexaminationrevealedvariationsinmucosalthickness,gobletcelldensity,andvillilengthacrossdifferentsamplinglocations.Inseabass,theintestinaltissuesfromKundMalir(Figure2A)displayedabalancedgobletcellcountwithwell-maintainedmucosalthicknessincontrast,theintestinaltissuesfromDam(Figure2B)showedmoderatemucosalthicknessandslightlyelongatedvilli.Figure2:ComparativeanalysisofintestinaltissuesofSeabass(Latescalcarifer)andSeabream(Nemipterusjaponicus)acrossvarioussamplingsitesTheseabasssamplesfromGadani(Figure2C)exhibitedreducedmucosalthicknessandanincreasednumberofGRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Page6gobletcells.Forseabream,theintestinaltissuesfromKundMalir(Figure2D)presentedthetallestvilliandwell-developedmucosalthickness.Incontrast,thesamplesfromDam(Figure2E)exhibitedmoderatemucosalthicknessandvillilength.TheintestinaltissuesfromseabreamcollectedatGadani(Figure2F)showedshortervilliandreducedmucosalthickness.Hematoxylinandeosinstainingoftheguttissuesofmajorcommercialfishseabass(Latescalcarifer)andseabream(Nemipterusjaponicus)ofdifferentlocationsintheBalochistancoastwereconducted.Inthehistology,variationsatthelevelofmucosalthickness,gobletcelldensity,andvilluslengthamongplacesofsamplingwereobserved.Gadaniintestinaltissueofseabass(Figure2A)revealedreducedmucosalthicknessandincreasedgobletcellnumber.Dam'sintestinaltissue(Figure2B)revealedmoderatemucosalthicknessandslightlyhypertrophiedvilli.SeabassfromKundMalir(Figure2C)revealedawell-balancednumberofgobletcellsandmaintainedmucosalthickness.Gadaniseabreamintestinaltissue(Figure2D)containsshortvilliandathinmucosa.Conversely,damsamples(Figure2E)containedintermediatevillilengthandmucosalthickness.IntestinaltissuesofseabreamatKundMalirrecovered(Figure2F)possessedthehighestvilliandwell-developedmucosalthickness.Table1:ShowstheassessmentofintestinalvillilengthinSeabassSeabassS.No12StudySiteKundMalirDamVillilength(µm)Significance43.89±7.9334.79±7.29NotSignificant(***p<0.0001)3Table1showssite-specificvariationsinintestinalvillilengthofLatescalcariferalongtheBalochistancoast.The(**p<0.001)Gadani36.13±8.46villiwerelongestatKundMalir(43.89±7.93µm),followedbyGadani(36.13±8.46µm)andDam(34.79±7.29µm).ThedifferenceswerehighlysignificantinfishfromDam(***p<0.0001)andGadani(p<0.001)comparedtoKundMalir.Table2:IndicatecomparisonofmucosallayerthicknessSeabassS.NO12StudySiteKundMalirDamMucosalthickness(µm)8.33±1.888.33±1.88SignificanceNodifferencep<0.053Table2presentsthecomparisonofintestinalmucosallayerthicknessinLatescalcarifer.ThemucosalthicknessNodifferenceGadani7.37±1.86wassimilaratKundMalir(8.33±1.88µm)andDam(8.33±1.88µm),whileaslightlyreducedvaluewasrecordedatGadani(7.37±1.86µm).Statisticalanalysisshowedasignificantdifference(p<0.05)atDamcomparedtotheothersites.GRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Table3:DisplaytheevaluationofintestinalvillilengthinNemipterusjaponicusSeabreamPage7S.No123However,thefindingsprovokedtoassociatedwithcollecteddatafromthreecoastalsitesalongtheBalochistanVillilength(µm)43.89±7.9334.79±7.2936.13±8.46SignificanceNotSignificant(**p<0.001)NotSignificantStudySiteKundMalirDamGadanicoast.ThevilliwereelongatedatKundMalir(43.89±7.93µm),followedbyGadani(36.13±8.46µm)andDam(34.79±7.29µm)(Table3).Statisticalanalysisrevealedasignificantdifference(p<0.001)infishfromDamcomparedtoKundMalir,whilethevariationatGadaniwasnotsignificant.Table4:IllustratethemucosallayerthicknessinSeabreamSeabreamS.NO123Table4illustratesthemucosallayerthicknessinNemipterusjaponicusfromstudysites.ThemucosalthicknessMucosalthickness(µm)8.33±1.887.11±1.677.37±1.86SignificanceNotsignificantp<0.05NotsignificantStudySiteKundMalirDamGadaniwashighestatKundMalir(8.33±1.88µm),followedbyGadani(7.37±1.86µm)andDam(7.11±1.67µm).Statisticalanalysisindicatedasignificantdifference(p<0.05)atDamcomparedtoKundMalir,whilenosignificantdifferencewasfoundatGadani.5.DISCUSSIONThecurrentstudyevaluatedtheintestinalhistologyof(Latescalcarifer)and(Nemipterusjaponicus)takenfromthreecoastalsitesinBalochistanDam,Gadani,andKundMalirtoidentifypotentialstructuraldifferences.SuchstudiesareveryrarefromthecoastofBalochsitan.Thereareseveralstudiesthathaveanalyzedtheseabasshistologyandphysiologicalparameter(Coz-Rakovacetal.,2005;Purushothamanetal.,2016;Thophonetal.,2003)&seabream(Kerdgarietal.,2013;Ibrahimetal.,2025).InthecurrentfindingsfishfromKundMalirshowedthelongestintestinalvilliandthethickestmucosallayer,reflectingahealthiergutcondition(Liuetal.,2025).Asimilarpatternwasdescribedinseveralstudiesunderexperimentalconditionswhofoundthatseabassexposedtominorchallenge(0.22ppmofheavymetal)hadlongvilliandathinmucosallayer.Fishexposedtodietarychangesshowedvillishorteningandtissuealterations.Likewise,Sklan,Prag,andLupatsch(2004)observedtallandwell-organizedvillilinedwithcolumnarepithelialcellsbearingabrushborderinhealthyTilapia.Inthesameway,Vélez-Calabriaetal.(2023),alsoreportedthatgiltheadseabream(Sparusaurata)fedwithhydrolyzedporcinemucosadevelopedthickervilliandmucosallayersthanthoseinthecontrolgroup(0dose),whichindicatesbetterintestinalstructureandnutrientabsorptioncapacity.FishcollectedfromDamandGadanishowedshorterintestinalvillilengthofseabass.asimilarobservationwasmadebyPedàetal.(2016),whofoundthatEuropeanseabass(Dicentrarchuslabrax)exposedtocontaminatedconditionsdevelopedshortenedandswollenvillialongwithepithelialdamage.TheirfindingsGRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Page8supportthepresentresults,indicatingthatunfavorableenvironmentalconditionscanleadtoweakenedintestinalstructureandreducedabsorptivecapacity.ThesefindingsmayrepresentourresultsfromGaddaniandDamsite.Thesesiteshavebeenwidelyusedforindustrialactivity.Pedàetal.(2016),speciesfrombothsitesshowedthinnermucosajustlike,Purushothamanetal.(2016),studywhoobservedabouta60%reductioninmucosalheightinstressedseabass(Latescalcarifer),indicatingweakenedintestinalhealth.FishfromDamandGadanishowedshortervilliandthinnermucosa,whilethosefromKundMalirhadahealthierintestinalstructure.Thismorphologicalcharacterpromoteseffectivedigestionandabsorptionoffeed.Inthecurrentstudy,thewell-preservedvilliatKundMalirmayindicatestablephysiologicalconditions.Besidesthis,shortervilliandthinnermucosallayersatDamandGadanimayreflectadaptiveorphysiologicalresponsestodifferencesthatcanbelinkedwithhabitats,eatinghabits,orphysicalvariable(temperature&salinity)stress.(Younisetal.,2013)observeddegenerationandshorteningofintestinalvilliinOreochromisniloticusexposedtosublethalcadmiumlevels.Al-Niaeem,Al-Hamadany,andAl-Tameemi(2010)observedvilliatrophyandmucosalthinninginCyprinuscarpioduringstarvationstress.Whilethecausesinpreviousinvestigations(Younisetal.,2013)weredeliberatelycreated,thetypeoftissuevariation(shortervilli)foundislikethemodificationsseeninthecurrentstudy.Thisshowsthatfishintestinesareextremelysusceptibletoenvironmentalorphysiologicalchanges,butitneedstobeanalyzedmoreindepthwithothermolecularapplications.Naturalhabitatchanges,suchaswatertemperature,salinity,orfoodavailability,whichareknowntoalterfishintestinalmorphology,couldexplaintheobservedvariationamongsites.Previousresearchhasfoundthatfishintestinestructureadjuststoeatingconditionsandhabitatfactors,influencingvilliformandmucosalthickness(Sklanetal.,2004).TheslightlyreducedvillilengthandmucosalthicknessseeninDamandGadanifishcouldbeattributedtonormalstructuralmodificationscausedbyenvironmentalconditionsratherthanpathogenicabnormalities.LatescalcariferandNemipterusjaponicushadcomparablepatternsofsite-wisevariation,howeverthedegreeofvariancedifferedeachspecies.Latescalcariferexhibitedmoresignificantvillishortening,butNemipterusjaponicushadslightlythinnermucosa,indicatinginterspecificheterogeneityinintestinalresponses.Thesedifferencescouldbeduetodietaryhabits,metabolism,orenvironmentalpreferences.Inconclusion,thehistologicalinvestigationdemonstratedthatintestinemorphologyvariedbetweenplacesalongtheBalochistancoast,presumablyduetonaturalenvironmentalchangesoradaptivephysiologicalresponses.ThemorphologicalparametersseenatKundMalirindicateacceptablehabitatconditions,whereasslightstructuralvariationsinfishfromDamandGadaniindicatepossiblesite-relatedimpacts.Thefindingsarecomparablewithpreviouslyrecordedintestinalfeaturesinteleosts,supportingthenotionthathistologicalpropertiesoffishintestinesaresensitivemarkersoftheirhealthandhabitatcircumstances(Al-Niaeemetal.,2010;Sklanetal.,2004;Younisetal.,2013).RegularhistologicalexaminationsofcommerciallyimportantfishspeciescanthusprovideusefulinformationaboutthebiologicalstateandhabitatqualityoftheBalochistancoastalecosystem.GRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Page96.CONCLUSIONThecurrentstudycomparesthehistologicalassessmentoftheintestineinLatescalcarifer(seabass)andNemipterusjaponicus(seabream)takenfromthreecoastalsitesinBalochistan.Thefindingsrevealedsignificantdifferencesinintestinalvillilengthandmucosalthicknessacrosssites.Overall,fishfromKundMalirhadlongervilliandthickermucosallayers,indicatingwell-developedintestinalsystems,whereasfishfromDamandGadanihadshortervilliandthinnermucosa.Thesedifferencesimplythatlocalenvironmentalorphysiologicalfactorsmayinfluenceintestineshapeintheseanimals.Althoughthespecificcausesofthesechangeswerenotinvestigatedinthecurrentstudy,theobservedstructuraldifferencescouldbenaturaladaptationstohabitatconditions,eatingpatterns,orotherphysiologicalinfluences.Theresultsshowthathistologicalparameterslikevilliheightandmucosalthicknesscanbeeffectiveindicatorsoffishintestines'overallhealthandfunctionality.Overall,thefindingsprovideessentialbaselineinformationontheintestinalhistologyoftwocommerciallyimportantfishspeciesfromtheBalochistancoast.Suchbaselinedataisvitalforfuturecomparativeandmonitoringstudiestounderstandhowenvironmentalandbiologicalfactorsinfluencefishtissueshapeandgeneralhealth.ACKNOWLEDGMENTOursincerethanksalsogototheFacultyofMarineSciencesandFacultyofAnimalScience,LasbelaUniversityofAgriculture,WaterandMarineSciences(LUAWMS),laboratorystafftoprovidethenecessaryfacilities,timeandresourcestocarryoutthisresearch.GRJNST,Volume:04-Issue2(2026)/ISSNP:2790-7643ArticleID:2064https://doi.org/10.53762/grjnst.04.02.15G.2064Page10REFRENCESAl-Niaeem,K.S.,Al-Hamadany,Q.H.,&Al-Tameemi,R.A.(2010).Histopathologicalchangesintheintestine,liverandpancreasofthecommoncarp(Cyprinuscarpio)duringstarvation.Proceedingsofthe6thInternationalConferenceonBiologicalSciences(Zoology),Uni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