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PGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205DOI:10.
3126/ijasbt.
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37Vol-2(2)June,2014PGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205DOI:10.
3126/ijasbt.
v2i2.
10352Thispapercanbedownloadedonlineathttp://ijasbt.
org&http://nepjol.
info/index.
php/IJASBTPHYSICOCHEMICALANALYSISANDMICROBIALDIVERSITYOFYAMUNAWATERANDINDUSTRIALEFFLUENTSPoonamGupta1,MonikaAsthana1,AvnishKumar*1andSiddharthaBarun21DepartmentofBiotechnology,SchoolofLifeSciences,KhandariCampus,Dr.
B.
R.
AmbedkarUniversityAgra2GujaratStateBiotechnologyMission,LeogeneProject,Gandhinagar-382017,Gujarat,India.
*Correspondence:avnishkumar81@gmail.
comAbstractPollutionhasarisenasaseriousenvironmentalconcerntothepresentworldafterindustrializationofhumansocieties.
Ithasseverelyaffectedourair,soilandwatersources.
Lookingtoitsglobal,national,regionalandlocaldimensions,itisnowimperativetocheckitateachandeverylevel.
Inthepresentstudy,8samples(3Yamunawatersamples,3tanneryeffluentsamplesand2textileeffluentsamples),werecollectedfromdifferentsitesofYamunaandexitpointsoftextileandtanneryIndustries.
Waterandeffluentsampleswereanalysedforvariousphysicochemicalparameters(pH,TDS,hardness,chlorideandBOD)usingconventionalmethods.
Afterwardsthesesampleswereutilizedforisolationofthenativebacterialspecies.
AllthesampleswereshowinghigherthanthestandardvaluesforTDS(500mg/l),hardness(80-100mg/l),chloride(250mg/l)andBOD(30mg/l).
ItwasobservedthatthetanneryeffluentswereshowingmaximumTDSvalues(1190-1240mg/l),followedbytextileeffluents(1190and1210mg/l)andYamunawater(530-1180mg/l).
Similarly,incaseofchloridecontent,highestconcentrationrange(828.
8-1598mg/l)wasshownbytanneryeffluents.
pHvaluewasnearlyneutralforYamunawater,slightlyacidicincaseoftextileeffluentsandmoreacidicfortannerysamples.
Highestrangeofhardnessvalueswereobservedforthetanneryeffluents(860-880mg/l)followedbytextileeffluents(760and860mg/l).
TheBODvalueswerenearlysimilarforallthesampleswithmaximumvaluesbeingobservedfortanneryeffluents(42-48mg/l).
Thusitcanbeinferredthatallthesampleswerehighlypollutedandneedtobetreatedbysuitablemethods.
Therewere11culturespurified,thatcouldbeemployedinbioremediationpurposes.
KeyWord:Biologicaloxygendemand(BOD);Chemicaloxygendemand(COD);Tannery;Textile;River;Totaldissolvedsolids(TDS).
IntroductionTheparamountofpollutioninourenvironmentespeciallyrivers,isadireconsequenceofcontinuallyexpandingpopulationalongwithanexponentialdevelopmentintheindustrialfield.
RiverYamuna,withatotallengthofaround1370kmisthelargesttributaryoftheGanges.
ThepollutionofriverYamunaisconsideredaninternationalissue.
Therearemanybigindustries,factories,andevenpeopleslivingincolonies,slumsandruralareasatthebankofYamuna.
Allthesearepollutingthisriver.
Amongthesecauses,industriesarecontributingmajorlytoincreasepollutionloadinriverYamunaespeciallytanneriesandtextileprintingunits.
ThetanningindustryplaysasignificantroleinIndianeconomybygeneratingexportsandemployment.
However,tannerywastesareamongthehighestpollutantindustrialwastes.
Tannerywasteisalwayscharacterizedbyitsstrongcolour(reddishdullbrown),highBOD,highpH,andhighdissolvedsolids.
Importantpollutantsassociatedwiththetanningindustryincludechlorides,tannins,chromium,sulphateandsulphidesasadditiontotraceorganicchemicalsandincreasinguseofsyntheticchemicalssuchaspesticides,dyesandfinishingagents,aswellasfromtheuseofnewerprocessingchemicalsolvents(Chandravathanam,2009).
Ithasbeenrecognizedtointerruptagro-basedactivitieslikefarmingandanimalhusbandryaswellaspolluteground-watersystems.
Groundwatercontaminationoccurswhenwastewaterandchemicalsseepthroughthesoilfromunlinedponds,pipesanddrains,orfromdumpsandspills.
Groundwatermaytakealongtimetocleanseitselfbecauseitmovesslowlyandisoutofcontactwithair.
Consequentlywhenthedepositionofsolidsisconsidered,itbecomesapparentthatrawunsettledtannerywastewaterscancauseencrustation(ofcalciumcarbonate)andseriouscorrosionofmetalsaswellasconcretesewers(duetoH2SbiologicaloxidationtoH2SO4)(Balusubramanianetal.
,2000).
Thushighpollutantloads,involvingchromium,sulphates,chloridesetccouldeasilyinterferewithkeybiologicalprocessesusedinsewagetreatmentplants.
Inadditionthesepollutantsmayalsodamagetheecologyofthereceivingterrestrialandaquaticsystemsinthevicinityofthedischargepoints(Mwinyihijaetal.
,2010).
BiologicalResearchArticlePGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205Thispapercanbedownloadedonlineathttp://ijasbt.
org&http://nepjol.
info/index.
php/IJASBTdecompositionoforganicmatterfromIndustrialwaste,aswellassulphideemissionsfromwastewaters(alsoduetobacterialreductionofsulphate)(Balusubramanianetal.
,2000),areresponsibleforthecharacteristicobjectionableodoursfromtanneriesintheformofH2S(Dormanetal.
,2000).
Essentiallyitisnowrecognizedthatdirectcontactwithsomeindustrialchemicalscanpotentiallycausedisability,illness(toxigenic/carcinogenic)andevendeathinhumans(Murtietal.
,1989).
Humanhealthcanalsobeaffectedbytoxichazardsthroughtheunskilledandunprotectedhandlingofpesticides,tanningchemicalsandtreatedhidesandskins(Mwinyihija,2011).
TextileindustryisoneoftheoldestindustriesinIndia.
Itisknownforusinglargequantitiesofwaterandvarietyofchemicals(Qinetal.
,2007).
Takingintoaccountthevolumeandcompositionofeffluent,thetextilewastewaterisratedasthemostpollutingamongallindustrialsectors(Robinsonetal.
,2001;Awomesoetal.
,2010;Vilasecaetal.
,2010;Babanetal.
,2010).
Ingeneral,thewastewaterfromatypicaltextileindustryischaracterizedbyhighvaluesofBOD,COD,colourandpH(YusuffandSonibare,2004;Tufekcietal.
,2007,).
Itisacomplexandhighlyvariablemixtureofmanypollutingsubstancesrangingfrominorganiccompoundsandelementstopolymersandorganicproducts(Brown,1983).
Incompleteuseandthewashingoperationsgivethetextilewastewateraconsiderableamountofdyes(Mathuretal.
,2005).
ConsideringtheseaspectsthepresentstudywasdesignedtoinvestigatethelevelofpollutioninYamunawateratAgraregionduetoselectedindustrialeffluents(tanneryandtextile).
Thephysicochemicalpropertieswasanalysedforthispurpose.
Thenativebacterialcultureswereisolatedfromthepollutedwaterandeffluentsamples,sothatitmaybeusefulforbiodegradationofvariousxenobioticcompounds(likevariousdyesandotherhumanmadenon-degradablecompounds)presentinwater.
MaterialsandMethodsPhysicochemicalanalysisofYamunawater,textileandtanneryeffluentsamplesSamplingwascarriedoutfrom8differentplaceslocatedinAgra,Mathura&Delhiasmentionedbelow:1.
Y1=PoiyaGhat,Agra(Yamuna)2.
Y2=Sikandarpur,Agra(Yamuna)3.
Y3=Khelgaon,NearDayalbagh,Agra(Yamuna)4.
Ta1=Tanneryfactory,GaonMehel,Runakta,Agra(Tannery)5.
Ta2=TanneryFactory,Runakta,Agra.
(Tannery)6.
Ta3=TanneryFactory,Agra(Tannery)7.
Tx1=TextileIndustry,NearJaiGurudev,Mathura(Textile)8.
Tx2=TextileIndustry,NewDelhi(Textile)Thewaterandeffluentsampleswerecollectedinsterileplasticbagsintwosets(Fig.
1).
Onesetwasutilizedforphysicochemicalanalysisandotherforbacterialisolation.
pHwasrecordedatthesamplingpoint.
Physico-chemicalanalysesofallthesamplesweredonebystandardmethods(APHA,1992).
Isolation&PurificationofbacteriafromYamunawater&effluentsamplesTheindigenousbacterialcultureswereisolatedseparatelyfromallthe8samplesbySpread-platemethodandthensubsequentlypurifiedbyrepeatedstreaking(QuadrantStreaking).
AllelevencultureswerethenallottedwithsuitableculturenumberandstoredinNutrientAgarslantsforfutureuse(Asthanaetal2014).
CharacterizationofthebacterialculturesThepurifiedbacterialcultureswerethencharacterizedonthebasisoftheircolonymorphology(Colour,Shape,ElevationandOpticalCharacteristics)andGram'sstaining(Asthanaetal2014)Fig.
1:MapofRiverYamunashowingsamplingpointsatAgraIndia.
PGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205Thispapercanbedownloadedonlineathttp://ijasbt.
org&http://nepjol.
info/index.
php/IJASBTResultsPhysicochemicalanalysisofYamunawater,textileandtanneryeffluentsamplesTheobservationsofthephysicochemicalanalysisarepresentedinTable1.
Outofthe8samplesanalysedonlysamples1stand2ndYamunawerecolourlesswithoutsediment,whileremainingwerecolouredwithsediment.
Samples1st,7thand8thwereodourlessandalltheothersampleswerehavingfoulodour.
IncomparisontothestandardvalueofT.
D.
S(500mg/L)allthesamples(530-1240mg/L)wereshowinghighervalues.
Itwasobservedthatthetanneryeffluents(1190-1240mg/l)wereshowingmaximumTDSvalues,followedbytextileeffluents(1190and1210mg/l)andYamunawater(530-1180mg/l).
Similarlychloridecontentofallthesamples(277.
9-1598mg/L)wasquitehighincomparisontostandardvalueofdrinkingwater(250mg/L)andmaximumvalueswereobtainedfortannerysamples(828.
8-1598mg/l)followedbytextilesamples(625.
7and628.
2mg/l)andYamunawater(277.
9-491.
3).
pHwasfoundtobenearneutral(6.
7-7.
5)forYamunawatersamples,slightlyacidic(6.
3&6.
5)incaseoftextileeffluentsandacidic(5.
1-5.
7)fortanneryeffluents.
ExceptforthetannerysamplespHoftheothersampleswasfoundtobewithinthepermissiblerangeofdrinkingwater.
AsincaseofotherparametersthehardnessandBODvaluesforallthesampleswerehigherthanthestandardvalues.
HighestrangeofHardnessvalueswereobservedforthetanneryeffluents(860-880mg/l)followedbytextileeffluents(860&760mg/l)andlastlyYamunawater(540-680mg/l).
TheBODvalueswerenearlysimilarforallthesampleswithmaximumvaluesbeingobservedfortanneryeffluents(42-48mg/l)andlowestvaluesobservedforYamunawater(31-35mg/l),whichisquiteclosertostandardvalue(30mg/l).
Table1:Physicochemicalparametersofthewater&effluentsamples:SampleIDLocationQuantitytakenColourandAppearanceOdourT.
D.
S(mg/L)Chloride(mg/L)pHvalueHardness(mg/L)B.
O.
D(mg/L)Y1Poiyaghat,Agra(Yamuna)500mLColourless,nosedimentationOdourless530277.
96.
754031Y2Bahadurpur,Khaspur,Agra(Yamuna)500mLColourless,nosedimentationFoulodour1160466.
57.
568033Y3Khelgaon,nearDayalbagh,Agra(Yamuna)500mLBlackishwithsedimentationFoulodour1180491.
37.
165035Ta1TanneryfactoryGaonMehel,Runakta,Agra(Tannery)500mlLightYellow,withdirtyblacksedimentationFoulOdour1230828.
85.
787542Ta2TanneryFactory,Runakta,Agra(Tannery)500mlBrown,withdirtyblacksedimentationFoulOdour124015985.
588048Ta3TanneryFactory,Agra(Tannery)500mlLightBrown,withdirtyblacksedimentationSlightFoulOdour119015885.
186045Tx1Textilefactory,Mathura(Textile)500mLBlackishwithsedimentationOdourless1210625.
76.
386038Tx2Textilefactory,NewDelhi(Textile)500mLBlackishwithsedimentationOdourless1190628.
26.
576035StdStandardvaluesfordrinkingwater-Colourless,nosedimentationOdourless5002506.
5-8.
580-10030Here:Y=Yamuna;Ta=Tannery;Tx=Textile;Std=StandardPGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205Thispapercanbedownloadedonlineathttp://ijasbt.
org&http://nepjol.
info/index.
php/IJASBTTable-2:SampledilutionandnumberandtypeofcolonypresentinYamunawater&TextileeffluentsamplesSampleIDDilutionNo.
ofcolonyTypeofcolonyTypeofculturespurifiedNameofpurifiedculturesY1(Yamuna)1*10-41011S.
1.
11*10-5--Y2(Yamuna)1*10-410022S.
2.
11*10-5--S.
2.
2Y3(Yamuna)1*10-46511S.
3.
11*10-5--Ta1(Tannery)1*10-420012S.
4.
11*10-5501S.
4.
2Ta2(Tannery)1*10-42411S.
5.
11*10-5--Ta3(Tannery)1*10-42011S.
6.
11*10-5--Tx1(Textile)1*10-43411S.
7.
11*10-5--Tx2(Textile)1*10-41912S.
8.
11*10-541S.
8.
2Here:Y=Yamuna;Ta=Tannery;Tx=TextileIsolation&PurificationofbacteriafromYamunawaterandeffluentsamplesSampleno.
4(tannery)andsampleno.
8(textile)wereshowngrowthat10-5dilution(Table2).
Intotal11bacterialcultureswerepurified(Table2).
Yamunawaterandtanneryeffluentsamplescontributedequalnumberofcultures(foureach)andtextileeffluentcontributedthreecultures.
CharacterizationofthebacterialculturesOnthebasisofcolonymorphologyitwasobservedthattheculturesfromYamunawaterwereeitherwhite,orangeorpaleyellowincolour,whereasculturesfromTanneryeffluentwerewhiteorcolourlessandculturesfromtextilewerewhiteororange(Table3).
Onthebasisoftheremainingparameters(shape,elevation&opticalcharacteristics)thecultureswereshowingvariablemorphology(Fig.
2).
AllfourbacterialculturespurifiedfromYamunawaterwereGramnegative(Fig.
3).
Ofthese,threewerecocciandonewasbacilli.
AlltheculturesfromTanneryeffluentwereGramnegativecocci(Table-3).
IncaseoftextileeffluenttwocultureswereGramnegativecocciandonewasGrampositivebacilli.
DiscussionThepresentstudywascarriedouttodeterminethevaluesofvariousphysicochemicalparametersinordertoevaluatethelevelofpollutionandtoisolatebacterialculturesfromYamunawaterandindustrialeffluents.
OnthebasisofphysicochemicalanalysisoftheYamunaandindustrialeffluentsamplesitwasobservedthattheTDSwerepresentatmuchhigherconcentrationinthetanneryandtextileeffluentsincomparisontoYamunawaterandstandardvaluefordrinkingwater.
SimilarlyGuptaetal.
(2012)reportedhighTDSValues(973mg/L–1664mg/L)fortextileeffluents.
HigherTDSvaluescanbeduetomanydissolvedsolidswhichmightbeundesirableforhealthofthereceivingwaterbody.
Dissolvedmineralsandorganicconstituentsmayproduceaestheticallydispleasingcolourandodour.
Alsothesolidsincreasetheturbidityofwater,inducesepticconditionsinthewaterbodybyretardingthephotosyntheticactivityandalsointerfacewithoxygentransfermechanismofair-waterinterface.
PGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205Thispapercanbedownloadedonlineathttp://ijasbt.
org&http://nepjol.
info/index.
php/IJASBTFig.
2:ColonyexaminationofpurifiedculturesFig.
3:Microscopicexaminationofpurifiedcultures:(A)Gramnegativecocci,(B)Gramnegativebacilli(C)GrampositivebacilliMaximumchloridecontentwasobservedintannerysamplesandthismightbebecauseofthevariouschlorinebasedchemicalsduringthesteps.
Nextinorderweretextilesamplesandthismightbebecauseofthevariouschlorinebasedchemicalsusedduringthebleachingsteps.
ChloridecontentofYamunawaterwasalsohigherthanthestandardvalueandthismightbebecauseitreceiveseffluentsfromvariousindustries.
AlltheYamunawatersamplesandtextileeffluentsamplewereshowingnearneutraltoslightlyacidicpH.
Allthetanneryeffluentsampleswereshowingacidicvalueanditmightbebecauseofhigherquantitiesofacidicchemicalsintanneryunits.
Hardnessvalueofallsampleswasmuchhigherthanthestandardvalue.
Itmightbeduetopresenceofcations(calcium,magnesiumandsodium)inthesamples.
Thisinturnmightbeduetotheuseofvariouschemicalsinthetannery,textileindustryandintheotherindustriesreleasingtheireffluentinYamuna.
ABCPGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205Thispapercanbedownloadedonlineathttp://ijasbt.
org&http://nepjol.
info/index.
php/IJASBTTable3:ColonymorphologyandGramstainingofisolatedbacterialculturesfromYamunawater,tanneryandtextileeffluentwaterCultureNameGram'sstainShape(Bacteria)ColourShape(Colony)ElevationOpticalCharacteristicsS.
1.
1(Y1)GramNegativecocciYellowSphericalConvexOpaqueS.
2.
1(Y2)GramNegativecocciOrangeSphericalConvexOpaqueS.
2.
2(Y2)GramNegativebacilliWhiteSphericalflatTransparentS.
3.
1(Y3)GramNegativecocciPaleyellowSphericalConvexOpaqueS.
4.
1(Ta1)GramNegativecocciWhiteSphericalConvexTransparentS.
4.
2(Ta1)GramNegativecocciWhiteSphericalflatOpaqueS.
5.
1(Ta2)GramNegativecocciColourlessSphericalConvexTransparentS.
6.
1(Ta3)GramNegativecocciWhiteIrregularflatTranslucentS.
7.
1(Tx1)GramNegativecocciWhiteIrregularflatOpaqueS.
8.
1(Tx2)GramNegativecocciwhiteIrregularflatOpaqueS.
8.
2(Tx2)GramPositivebacilliOrangeSphericalConvexOpaqueHere:Y=Yamuna;Ta=Tannery;Tx=TextileItwasobservedthattheBODvaluesofYamunawateraswellastheindustrialeffluentswerehigherthanstandard.
Thispointtowardstheoxygendeficiencyinthewaterbodiesduetodischargeofindustrialeffluents,richinorganiccompounds.
Highvaluesofthisparameterindicatepotentialdepletionofdissolvedoxygeninthewaterbody.
Deficiencyofoxygeninreceivingwatercouldcauseadverseeffectsonaquaticlife.
Inworstcase,thiscanresultintotaldepletionofoxygeninreceivingwater,causingananaerobicenvironment,thuschangingthehabitatfromaerobictoanaerobiclife.
ThephysicochemicalanalysisoftheYamunawaterandtheeffluentsgivesanideaabouttheseverity,typeandpossiblesourceofpollution,andcanbeusedasanargumenttoemphasizeonthetreatmentofYamunaaswellastheindustrialeffluentspriortotheirdischargeontheopenlandorlocalwaterbodies.
Aftercompletionofpurificationstep11bacterialcultureswereobtained,whichcanbetermedasnativemicrofloraoftheYamunaandindustrialeffluentsamplesasnogrowthwasobtainedoncontrolplatesofNutrientagar,andthisruledoutanypossibilityofthelaboratorycontamination.
Yamunawater,tanneryandtextileeffluentscontributednearlyequalnumberofbacterialcultures(4,4and3culturesrespectively).
Presenceoftheseculturesinthesamplessuggeststhattheyareadaptedtotheirpollutedenvironment.
InasimilarstudybyGuptaetal.
(2013)isolationof26bacterialculturesfromtextileeffluentshasbeenreported.
The11bacterialculturesobtainedfromYamunawaterandeffluentswerecharacterizedonthebasisofcolonycharacteristicsandGram'sreaction.
WhentheisolateswerecomparedonthebasisofcolonycharacteristicsitwasfoundthatsomeisolatesfromYamunawereshowingquitedifferentcolonycharacteristics,butmostoftheisolatesfromtanneryandtextileeffluentswerefoundtobesimilar.
ThisresultindicatesthatthereismorediversityinthebacterialmicrofloraofYamunaduetodilutionoftheindustrialeffluents.
ItwasobservedthatisolatesfromtheYamunaandeffluentswereshowingquitedifferentcolonycharacteristics,whichmeansthatisolatesobtainedfromYamunamightbedifferentfromtheisolatesofeffluents.
YamunawatercontributedmajorlyGramnegativecocciandonlyoneGramnegativebacilliwhiletanneryeffluentcontributedonlyGramnegativecocci(Vermaetal2011).
MajorityoftheculturesobtainedfromtextileeffluentwerePGuptaetal.
(2014)IntJApplSciBiotechnol,Vol2(2):199-205Thispapercanbedownloadedonlineathttp://ijasbt.
org&http://nepjol.
info/index.
php/IJASBTGramNegativecocciwhileonlyoneculturewasGrampositivebacilli.
ItmightbeconcludedthatGramnegativecocciismoreadaptedtogrowinthepollutedenvironmentandthuscanbeaverypotentcandidateinthefieldofbioremediation.
AcknowledgementAuthorsarethankfultoDean,SchoolofLifeSciencesandHead,DepartmentofBotany,SchoolofLifeSciences,BhimRaoAmbedkarUniversity,Agra,forconstantsupportduringthework.
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