BACTERIOLOGICAL AND PHYSICOCHEMICAL ANALYSIS OF WASTE WATER FROM FISH PONDS

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1 Ethiopian Journal of Environmntal Studis & Managmnt 9 (2): , ISSN: doi: Submittd: Novmbr 04, 2015 ccptd: March 03, 2016 BCTERIOLOICL ND PHYSICOCHEMICL NLYSIS OF WSTE WTER FROM FISH PONDS *SULE, I.O., BBIK, T.O., HMED, R.N., SLIU, B.K. ND OLYINK, K. J. Dpartmnt of Microbiology, Faculty of Lif Scincs, Univrsity of Ilorin, PMB 1515 Ilorin, Nigria bstract Raring of fish spcially catfish is vry common in many communitis in Nigria and th wastwatr from ths fish ponds ar oftn dischargd into th surrounding drains. ssssmnt of th bactriological and physicochmical charactristics of tn frshwatr fish ponds stockd with frican catfish (Clarias garipinus) was conductd within Ilorin mtropolis. Th rsults of physicochmical proprtis of th watr sampls showd that th ph, total hardnss, salinity, and suspndd solid rangd from , mg/l, g/l, and g/100ml rspctivly. Th bactrial count, total coliform, and facal coliform count of th wastwatr rangd from 4.0 x x 10 5 cfu/ml, nil to 2 x 10 4 cfu/ml, and nil to 1 x 10 3 cfu/ml rspctivly. Th bactrial spcis isolatd from th ponds wr Eschrichia coli, Staphylococcus sp., Staphylococcus aurus, romonas hydrophila, Staphylococcus saprophyticus, Chromobactrium sp, Micrococcus sp., Micrococcus kristina, Providncia sp., and Listria sp. Th antibiotics suscptibility tsting of th bactrial isolats wr dtrmind using th disc mthod. ll th isolats xhibitd multipl antibiotic rsistanc (MR). It can b concludd from this study that thr is nd to monitor th quality of wastwatr from th fish ponds bfor bing dischargd into th nvironmnt sinc potntial pathogns wr isolatd and most of ths isolats wr multipl antibiotic rsistant. This study is of socioconomic significanc in th sns that whil th farmrs ar abl to rais thir fish in th ponds and driv thir livlihood, thy ar bing cautiond on th nd to trat thir wast watr bfor bing dischargd into th rciving watr body. Ky Words: Wastwatr, Fish pond, Catfish, ssssmnt, ntibiotic rsistanc Introduction Wastwatr compriss liquid wast such as swag, oil and chmicals. Ths wast watrs contain a wid rang of potntial contaminants. Wast watr contains offnsiv and potntially dangrous substancs which ar mostly of anthropognic origin and causing pollution and contamination of rciving watr bodis (Ikpi and Offm, 2011). Fortunatly, watr can naturally rnw and clans itslf via various physical mthods: dilution, sunlight, aration, flocculation, sdimntation, biological dgradation *Corrsponding uthor: Sul, I.O. sulism@gmail.com 167 (dcomposition), prdatory activitis of microorganisms tc (Hanlor, 2013). This procss of rnwal is calld slfpurification and larg bodis of watr is said to possss slf-purifying capacity (Bitton, 2005). Contamination of watr is a srious nvironmntal problm as it affcts th diffrnt uss of th watr. Th us of indicator bactria such as facal coliforms in watr quality dtrmination is widly applid. Coliforms and Eschrichia coli ar of grat importanc among bactrial indicators usd in watr quality assssmnt (Torimiro t al., 2014).

2 Bactriological and Physicochmical nalysis of Wast Watr...SULE t al. On of th prominnt uss of surfac watr such as rivr, pond, run-off tc is in aquacultur which has contributd immnsly to th food sctor and rducd th risk of fishs at th brink of xtinction du to xcssiv catch in laks and ocans (Tidwll and llan, 2001). Mor than onfourth of all animal protin consumd by man is aquatic in origin (FO, 2000). quacultur has bn practicd in many sian countris for cnturis, but is a nw form of agricultur in many frican and Latin mrican countris. It is dfind as th cultivation of animals and plants in aquatic nvironmnts. quaculturists manipulat crtain componnts of th nvironmnt to achiv gratr control on production of aquatic organisms (Purdom, 1996). In Nigria, th raring of frican catfish (Clarias garipinus) is hlping in th provision of food and its usfulnss cannot b ovr-mphasizd as th dmand for fish is xpanding rapidly throughout th world (Ugwuba and Chukwuji, 2010). Thr is an intricat rlationship btwn th fish, thir biotic and abiotic nvironmnts and changs in on componnt may rflct and affct th othr (Wurt, 2000). Pond watr or watr usd in intnsiv fish raring contain many microorganisms which could b pathognic or opportunistic pathogn to fish, human, and planktons (Zmyslowska t al., 2003). Ths contaminating microorganisms hav bn attributd to qustionabl watr quality which can b tracd to th watr sourcs and high stocking dnsitis (Okpokwasili and kujobi, 1996). Watr sourcs for arthn ponds ar usually untratd surfac watr runoffs from strams, rivrs, laks, stord watrs whil undrground watr sourc is bing utilizd for most concrt ponds. 168 Watr from undrground sourcs such as a wll is pumpd and stord in storag tank which is usd to rfill th ponds whn th watr is to b changd at intrvals of about 3-5 days. Th fd usd for fish in ths ponds contain organic matrials and introducs a wid varity of microorganisms into th ponds. Omojowo and Omojasola (2013) isolatd six bactrial pathogns: E. coli, E. coli 0157:H7, Shiglla dysntria, Staphylococcus aurus, Salmonlla typhi, and romonas hydrophila from Cow dung usd as fd in th fish pond. Th microbial flora of a cultivatd fish is a rflction of its aquous nvironmnt (Sugita t al., 1985; Oni t al., 2013). Frshwatr fish in ponds commonly suffr from bactrial disass such as various kinds of skin ulcrations, albinodrma, rythrodrma, furunculosis, and vrticl-scal disas, primarily causd by romonas sp. and Psudomonas sp. (Das, 2004). Som of ths disass wr rportd to b most svr during th dry sason, whn dclining watr quality is a problm. In rizona, significant mortalitis of culturd tilapia rsultd from infction with a salt-tolrant strain of th bactrium romonas hydrophilia (Llobrra and acutan, 1987). Som prvalnt pathognic bactria of milkfish and tilapia isolatd in Taiwan includ spcis of th gnra: romonas, Edwardsilla, Flavobactrium, Psudomonas, and Strptococcus (Sursh and Lin, 1992). ricktssia-lik bactrium has also bn idntifid as a causativ agnt for disas outbraks in tilapia culturd in Taiwan. Th sprad of multipl antibioticrsistant pathognic bactria hav bn rcognisd by th World Organization for nimal Halth (WOH), th Food and gricultur Organization (FO) and th

3 Ethiopian Journal of Environmntal Studis and Managmnt Vol. 9 no World Halth Organization (WHO) as a srious global human and animal halth problm. Th dvlopmnt of bactrial antimicrobial rsistanc is nithr an unxpctd nor a nw phnomnon. It is, howvr, an incrasingly troublsom situation bcaus of th frquncy with which nw mrging rsistanc phnotyps ar occurring among many bactrial pathogns and vn commnsal organisms. Rsistanc has bn obsrvd to ssntially all of th antimicrobial agnts currntly approvd for us in human and vtrinary clinical mdicin. Th scop of this rsarch is to lucidat th bactrial contaminants in wast watr from fish ponds as wll as thir antibiotics suscptibility pattrns. It is common to s many fish ponds within our community raising diffrnt typs of fish spcially catfish. Th wast watr from ths fish ponds ar oftn dischargd into th surrounding drains. Hnc, this study is conductd to dtrmin th quality of this wast watr in ordr to protct th nvironmnt, plant, animal and man. Th aims and objctivs of this rsarch wr to dtrmin th physicochmical and bactriological charactristics of wast watr from fish ponds; isolat, idntify and dtrmin th occurrnc of th bactrial spcis; and th antibiotics suscptibility pattrns of th bactrial isolats. Mthodology Study ra Wastwatr sampls wr collctd from som fish ponds within Ilorin mtropolis. Ths wr 8 arthn ponds: TJ fish farm I, sa dam I, Tank, Ilsanmi, Osr I, TJ fish farm II, sa dam II, and Osr II; and 2 concrt ponds:.r., and Phonix. 169 Sampl Collction Th wastwatr was collctd asptically into a stril ambr bottl at a dpth 10 15cm from th surfac of th watr (Njoku t al., 2015). Th watr sampls wr transportd in black polythn bag immdiatly for analysis. Th sampling was don during th months of Novmbr 2014 to January 2015 btwn 8 10 am. Dtrmination of ph Th ph of th watr sampl was dtrmind using a digital Dnvr ph mtr modl 20 with glass lctrod. This was don according to standard mthod (PH, 1998). Dtrmination of Suspndd Solid This was dtrmind by filtration mthod using Whatman s filtr paprs (Duby and Mahshwari, 2005). Dtrmination of Salinity Contnt of th Watr Sampls This was dtrmind using argntiomtric titration according to BP, Dtrmination of Total Hardnss This was dtrmind using complxomtric titration as dscribd by Duby and Mahshwari (2005). Bactriological nalysis Th bactrial count of th wastwatr was dtrmind using pour plat tchniqu whil sprad plat mthods wr usd for th total and facal coliform counts. Th mdia usd wr nutrint agar, MacConky agar, and osin mthyln blu agar for bactrial count, total coliform, and facal coliform count rspctivly (Fawol and Oso, 2007; Willy t al., 2011). Charactrization and Idntification of Bactrial Isolats Th isolatd bactria wr charactrizd basd on thir colonial morphology, cllular morphology and

4 Bactriological and Physicochmical nalysis of Wast Watr...SULE t al. biochmical proprtis and thn idntifid by making rfrnc to standard txtbooks (Mac Faddin, 1980; Cowan and Stl, 1985). ntimicrobial Suscptibility Tst Th broth cultur of ach isolat was standardizd using 0.5 McFarland standard. Thn, st plat of stril Mullr Hinton agar was inoculatd with th standardizd tst organism and th antibiotic disc was placd on th inoculatd mdium. Th mdium was incubatd at 37 o C for 24 hours. ftr incubation, th plat was obsrvd for zons of inhibition around ach of th antibiotics. Th diamtr of th zon of inhibition was thn masurd in millimtr (mm) (CLSI, 2005; Brown and Macowan, 2010). ram positiv antibiotic disc was usd for ram positiv organism whil ram ngativ antibiotic disc was usd for ram ngativ organism. Multipl ntibiotics Rsistanc (MR) Indxing of Isolats Th multidrug rsistanc of th bactrial isolats wr analyzd using th multipl antibiotic rsistanc (MR) indx. Th multipl antibiotic rsistanc (MR) indx was dfind as a/b whr a rprsnt th numbr of antibiotics to which th isolat was rsistant to and b th numbr of antibiotics to which th isolat was xposd (Chitanand t al., 2010). Data nalysis Th data wr analysd using SPSS 15.0 packag. ll data ar man of thr rplicats. Th man, rang and standard dviation of ach paramtr was dtrmind. Th mans wr sparatd using Duncan s Multipl Rang tst (SPSS, 2010). Rsults Physicochmical Paramtrs of Wast Watr from Fish Ponds Th physicochmical proprtis of th wastwatr showd a ph rang of , total hardnss mg/l, salinity contnt g/l, and suspndd solid g/100ml (Tabl 1). Bactriological Counts of Wast Watr from Fish Ponds Th bactrial count, total coliform and facal coliform of th wastwatr rangd from 4.0 x x 10 5 cfu/ml, nil to 2 x 10 4 cfu/ml, and nil to 1 x 10 3 cfu/ml rspctivly (Tabl 2). Bactrial isolats and thir occurrnc Tn bactria spcis wr idntifid (Tabl 3) and thir occurrncs was as prsntd in Tabl 4. This compriss both ram positiv and ram ngativ bactria. ntibiotic suscptibility tsting (ST) of bactrial isolats ll th ram positiv bactria wr suscptibl to Ofloxacin but rsistant to Cftazidim, Cfuroxim, moxycillin- Clavulinat antibiotics. Similarly, all th ram ngativ bactria wr suscptibl to Nitrofuratoin, Ofloxacin, Ciprofloxacin, but rsistant to moxycillin-clavulinat, mpicillin, Cfuroxim, Cftazidim antibiotics (Tabl 5). ll th isolats showd multipl antibiotic rsistanc (MR) (Tabl 6). Th isolats with MR indx valus highr than 0.2 wr considrd multipl rsistant (Chitanand t al., 2010). 170

5 Ethiopian Journal of Environmntal Studis and Managmnt Vol. 9 no Tabl 1: Physicochmical charactristics of wastwatr from th fish ponds Sampl ph Salinity contnt (g/l) Suspndd solid (g/100ml) Total Hardnss (mg/l) T J fish farm I * 8.02 g ± f ± g ± ±4.0 T J fish farm II * 8.43 h ± f± b ± d ±5.0 Ilsanmi * 7.34 ± bc ± c ± cd ±4.0 Tank * 7.25 d ± bc ± d ± d ±5.0.R. ** 8.04 g ± b ± a ± a ±5.0 Phonix ** 8.04 g ± a ± a ± a ±6.0 Osr I * 6.77 a ± f ± f ± b ±4.0 sa dam I * 7.46 f ± cd ± c ± bc ±5.0 Osr II * 7.19 c ± d ± ± bcd ±4.0 sa dam II * 6.83 b ± cd ± ± cd ±4.0 *, Earthn pond; **, Concrt pond Man followd by th sam suprscript within th sam column ar not significantly diffrnt at α = 0.05 basd on Duncan s multipl rang tst (DMR). Tabl 2: Bactriological counts of wastwatr from th fish ponds Sampl Bactrial count (cfu/ml) x 10 3 Total coliform count (cfu/ml) x 10 3 Facal coliform (cfu/ml) T J fish farm I * 300 f ±20 20 c ±2 0 a ±0 T J fish farm II * 200 ±10 0 a ±0 0 a ±0 Ilsanmi * 13 a ±2 0 a ±0 0 a ±0 Tank * 4.0 a ±1 0 a ±0 0 a ±0.R. ** 66 b ±3 0 a ±0 0 a ±0 Phonix ** 78 bc ±4 0 a ±0 0 a ±0 Osr I * 180 d ±20 0 a ±0 0 a ±0 sa dam I * 19 a ±2 2.0 b ±0 1.0 b ±0 Osr II * 670 g ±20 0 a ±0 0 a ±0 sa dam II * 89 c ± b ±0 1.0 b ±0 *, Earthn pond; **, Concrt pond Man followd by th sam suprscript within th sam column ar not significantly diffrnt at α = 0.05 basd on Duncan s multipl rang tst (DMR). 171

6 Tabl 3: Charactrization and idntification of bactrial isolats Bactrial isolats Colonial colour Colonial shap ram s raction Cll s shap Clls arrangmnt Motility Catals Coagulas Indol Oxidas Mthyl rd Vogs Proskaur Citrat Uras Starch hydrolysis TSI 1 Cr Ir - C Cl k + F + romonas hydrophila 2 W Ci + B S F - Micrococcus sp. 3 y Ci + C Cl F - Micrococcus kristina 4 Cr Ir + C S F - Staphylococcus aurus 5 W Ci + C Cl F - Staphylococcus sp. 6 W Ci + C Cl F - Staphylococcus saprophyticus 7 y Ci + B S F + Listria sp. 8 Cr Ci - C Cl F - Chromobctrium sp. 9 Cr Ci - B Cl F - Providncia sp. 1 0 Cr Ci - B Cl F - Eschrichia coli Ky: Cr = cram; W = whit; y = yllow; Ci = circular; Ir = irrgular; ;B = bacilli; C = cocci; S = Singl; Cl = clustr F = Frmntativ; - = ngativ raction; + = positiv raction; OF = Oxidation-Frmntation; K = cid/lkalin raction; = cid/cid raction; = cid and gas; = cid only NO3 rduction Maltos Sucros Lactos Mannitol lucos Fructos OF H2S Probabl idntity 172

7 Bactriological and Physicochmical nalysis of Wast Watr...SULE t al. Tabl 4: Occurrnc of bactrial isolats in wastwatr from th fish ponds Bactrial isolats T J fish farm I T J fish farm II Ilsanmi Tank.R. Phonix Osr I sa dam Osr II sa dam II romonas hydrophila Micrococcus sp Micrococcus kristina Staphylococcus aurus Staphylococcus sp Listria sp. + Chromobactrium sp Staphylococcus saprophyticus Providncia sp Eschrichia coli + + Ky: +: isolatd : not isolatd Tabl 5: ntibiotics suscptibility pattrn of bactrial isolats +, prsnc of inhibition; -, absnc of zon of inhibition; ( ), diamtr of zon of inhibition (mm); CZ, Cftazidim 30µg; CRX, Cfuroxim 30µg; EN, ntamicin 10µg; OFL, Ofloxacin 5µg; U, moxycillin-clavulinat 30 µg; CTR, Cftriaxon 30µg; CPR, Ciprofloxacin 5µg; ERY, Erythromycin 5µg; NIT, Nitrofurantoin 300µg; CXC, Cloxicillin 5µg; MP, mpicillin 10 µg 173

8 Ethiopian Journal of Environmntal Studis and Managmnt Vol. 9 no Tabl 6: Multipl antibiotics rsistanc (MR) of th bactrial isolats Bactrial isolats Numbr of antibiotics rsistant to (a) Numbr of antibiotics usd (b) MR Indx (a/b) romonas hydrophila Micrococcus sp Micrococcus kristina Staphylococcus aurus Staphylococcus sp Listria sp Chromobactrium sp Staphylococcus saprophyticus Providncia sp Eschrichia coli Discussion Eighty prcnt of th fish ponds usd in this study wr arthn ponds. Most of th concrt fish ponds wastwatr hav highr ph (tnd to b alkalin) whn compard with th arthn ponds wastwatr. This could b du to dissolution of ions from th soil into th fish pond s watr. Th optimal watr hardnss ncssary for fish to thriv is dpndnt on th spcis of fish. Most fish grow wll ovr a wid rang of hardnss valus. Th total hardnss valus in this study is btwn mg/l and falls within th rcommndd standard of mg/l for fish cultur and within th prmissibl 100mg/l hardnss for catfish (Dpartmnt of Watr ffairs and Forstry, 1996). Th arthn ponds hav highr lvl of suspndd solid whn compard to concrt ponds which is du to th soil sdimnts, disturbancs whn watr is to b changd, agitation by th fish during fding and a hungr protst. Earthn ponds mimic th natural habitat of ths fishs with charactristics which includ dirct xposur to sunlight, prsnc of soil bnath th watr, prsnc of othr aquatic plants.g. zooplankton, alga, 174 diatoms, phytoplankton, watr dris owing to natural causs (vaporation). Salinity is among th most important factors and xrts various ffcts on th vitality of microorganisms. Th diffrncs in th salinity ar attributd to th incras in th vaporation rat (Torimiro t al., 2014). Salinity contnt in arthn ponds wr slightly highr owing to incrasd vaporation rat compard to that in concrt ponds and som of th watr in arthn ponds infiltrat into th soil rsulting into watr loss. Th rsult of th bactriological analysis showd that th bactrial count varid with location and this may b du to diffrncs in managmnt practics rsulting in diffrnt lvl of organic loads in th pond systm from th dit usd in fding th fish (Oni t al., 2013). Th sourc of microorganisms could also b tracd to sourcs of watr fd into th pond. Wll watr is xpctd to b of good quality whn usd for th pond than watr from rivr surfac runoff or stram. lso th frquncy of chang of th watr spcially in concrt ponds influncs th microbial load. Concrt ponds ar compltly dischargd and rfilld with frsh watr whil arthn ponds uss a topping systm whr mor watr is addd in ordr to maintain spcific volum or

9 Bactriological and Physicochmical nalysis of Wast Watr...SULE t al. watr lvls. It was obsrvd that th microbial load in arthn ponds wr considrably highr than that of concrt ponds. In a similar study, Nzh and Udz (2012) obtaind bactrial counts which rangd from 1.0 x x10 4 cfu/ml and 9.0 x x10 4 cfu/ml in arthn and concrt ponds rspctivly. Th prsistnc of pathogns in th watr nvironmnt is considrd as on of th crucial factors for infction transmission in trms of acut outbraks of disas. This study rvald th prsnc of Eschrichia coli, romonas hydrophila, Micrococcus kristina, Micrococcus sp., Staphylococcus sp., Staphylococcus saprophyticus, Staphylococcus aurus, Listria sp., Chromobactrium sp., and Providncia sp. in th fish ponds watr and this may pos a thrat to th halth of th fishs and consumrs. It was found that ram ngativ bactria gnra prdominat and this is in agrmnt with findings rportd by Molokwu and Okpokwasili, (2002), kinwal t al. (2007), Khunthongpan t al. (2013), and Njoku t al. (2015) in thir study on th divrsity of microflora of fish pond watr. Furthrmor, Elsaidy t al. (2015) isolatd E. coli, coliforms and Salmonlla sp. from watr and fish bing fd with raw and frmntd manur. Th rcovry of Eschrichia coli in two of th fish ponds could suggst a possibl facal contamination of th fish ponds. Th facal matrial may b as a rsult of frtilization of th ponds with animal manur which is dischargd dirctly into th fish ponds, or xcrtd by th fish into th ponds (yulo t al., 1994; Omojowo and Omojasola, 2013). ll th organisms wr suscptibl to Ofloxacin which can b usd for trating infctions in both humans and animals at appropriat concntrations. In a similar study, Morrison (2006) dtctd oxacillin 175 rsistanc in Staphylococcus sciuri group. Th dtction of pathognic bactria that ar multi-drug rsistant that is having multipl antibiotics rsistanc (MR) indx valus highr than 0.2 call for th nd for formulation and implmntation of cod of practic for fish farmrs. This is to nsur appropriat fish managmnt and prvntion of transmission of potntial pathogns which is important in th part of food safty plan. Conclusion It can b concludd from this study that thr is nd to monitor th quality of wastwatr from th fish ponds bfor bing dischargd into th nvironmnt sinc potntial pathogns wr isolatd and most of ths isolats wr multipl antibiotics rsistant. Rcommndation It is rcommndd that microbiological analysis and physicochmical xamination of wastwatr or fish for signs of possibl infctions should b conductd on a rgular basis. Th ministry of agricultur should nsur that th fish farmrs ar supplid with halthy fry for thir stock. ood quality watr such as wll or borhol should b usd in th fish pond rathr than watr from qustionabl sourcs such as rivr, stram, and surfac runoff. Th fish fds should b sourcd from rputabl manufacturrs. Watr in th fish pond should b changd compltly at rgular intrvals. Concrt ponds should b usd for raring of fish rathr than arthn ponds. Wast watr should b tratd ithr by physical mthods which involv filtration through slow sand filtrs, rapid sand filtrs, sand bds or chmical mthods such as addition of disinfctants g chlorin bfor final disposal into surrounding drains.

10 Ethiopian Journal of Environmntal Studis and Managmnt Vol. 9 no cknowldgmnts Th authors acknowldg th coopration rcivd from th ownrs of th fish ponds whr watr sampls wr collctd. Rfrnc kinwal, M.M.., Kk, R.I. and Egnonu, C. (2007). Bactrial microflora of frican catfish (Clarias garipinus) juvnils raisd undr fish cum pig intgratd farming systm in frsh watr arthn ponds during th dry sason. Nig. J. Fishris, 4(2): PH (1998). Standard Mthods for th Examination of Watr and Wastwatr, 20 th dition. mrican Public Halth ssociation, Washington DC, ISBN: yulo,.m., Machado, R.. and Scussl, V.M. (1994). Entrotoxignic Eschrichia coli and Staphylococcus aurus in fish and safood from th southrn rgion of Brazil. Int. J. Food Microbiol., 24: Bitton,. (2005). Wast watr Microbiology, 3 rd dition, John wily and Sons, Inc., Hobokn, Nw Jrsy, US. P. 400 BP (1993). British pharmacopia, volum I, Intrnational dition, HMSO publication cntr, London pp Brown, D. and Macowan,. (2010). Harmonization of antimicrobial suscptibility tsting brakpoints in Europ: implications for rporting intrmdiat suscptibility. J. ntimicrob. Chmothr., 65: Chitanand, M.P., Kadam, T.., yananath,., Totwad, N.D. and Balhal, D.K. (2010). Multipl antibiotic rsistanc indxing of coliforms to idntify high risk contamination sits in aquatic nvironmnt. Indian Journal of Microbiology 50(2): Doi: /s CLSI (2005). Prformanc standards for antimicrobial suscptibility tsting. Fiftnth informational supplmnt, M100-S15, vol. 25, No. 1.Clinical and Laboratory Standard Institut, Wayn, Pa. Cowan S.T. and Stl, K.J. (1985). Manual for th idntification of mdical bactria. 4 th dition, Cambridg Univrsity Prss, London. 217pp. Das, B.K. (2004). Bactrial disass of fish and thir control. In: Swain P., Sahoo P.K. Das B.K. Disas Diagnosis and Halth Managmnt of Frshwatr Fish and Shllfish. Bhubanshwar, Cntral Institut of Frshwatr quacultur, Dpartmnt of Watr ffairs and Forstry (1996). South frican Watr quality uidlins, gricultural us: quacultur 2 nd Edition 6, 170pp. Duby, R.C. and Mahshwari, D.K. (2005). Practical Microbiology. S.Chand and company Ltd., Nw Dlhi. 352pp. Elsaidy N., boulnnin, F. and Kirlla,..K. (2015). Impact of using raw or frmntd manur as fish fd on microbial quality of watr and fish. Th Egyptian Journal of quatic Rsarch 41(1): FO (2000). Th Stat of World Fishris and quacultur. Food and gricultural Organization, Rom, Italy. Fawol, M.O. and Oso, B.. (2007). Laboratory Manual of Microbiology. Spctrum Books Limitd, Ibadan, 123 pp. 176

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