1、API RECOMMENDED PRACTICE For API RPS8 Tfdrd F.cUtion December 1975 BIOLOGICAL ANALYSIS OF SUBSURFACE INJECTION WATERS OPFICIAI.PlaUCATION 111.11,u.-.PAIIIRT OPlICS AMERICAN PETROLEUM INSTITUTE Washington,D.C.20037 Issued by AMERICAN PETROLEUM INSTITUTE Production Department 300 Corrigan Tower Buildi
2、ng Dallas,Texas 7 5201 uopyrlght C 1976 by American Petroleum Institute 4 American Petroleum IDBt.itute General Bacterial Counts of Injection Waters 10.The following medium shall be uaed for making bactei-ial counts on waters containing less than 20,000 ppm(mg/1 liter)total solids:Beef extract,grams
3、.-8.0 Teyptone,grams-6.0 J)extroee,grlUlIS-1.0 Agar,grams.15.0 Distilled water,milliliters i.,000.0 11.Adjust pH to 7.0 with sodfum hydroxide(NaOH)and sterilir.e at 15 lb steam pressu1,e for 15 min.12.The sulfate-reducing bacteria mediwn(Par.18)shall be used for making plate counts cm wate18 contain
4、ing over 20,000 ppm total solids.13.Pour plat.ea are more desirable tha.n spreadplates becall88 of minimization of surface spreading colonies.Dilutions and plate counts are to be made in the mannei:reirenoed by the lat.est edition of S?Methads fr,t the Ea:ammation of Watet and Wa.re W aur.t 14.Plate
5、s shall be incubated under aerobic conditions and within 5 C of the recorded tempelature the water when 11ampled.15.Plates should be observed for growth after 2to 5 daY)l incubation.16.Fo untreated wate18,bacterial counts of lessthan 10,000 organisms per milliliter are genei-ally of little signjflca
6、nce.The significance of cowits above 10,000 per milliliter will aepend upon additional evidence such as loss of injectivity,incleased wellhead JlleSSure_ or filter plugging.17.A suitable broth cultu1-e method for aerobicbacteria is p1888nted as Appendix A,Section A-II,Par.A2.Sulfate-redacfng Bacteri
7、a Counts 18.A medium of the followhlg compmition shallbe used for counting sulfate-reducing bacteria:Sodiwn lactate,USP,milliliters-4,0 Yeast extract,grams-LO Thb medium-:,be i,urchued in the deb:,dratecl form ea TGEApr.tATllllable from American Pobllo-Health A.No,Ino.,1190 BlOlld,.-N.Y,N,Y.14112,IB
8、ulfatie-rednolne ba.cteda medium oan be obtained In dehydratedConn frlxn the biological auwb hou.Ascorbic acid,grams-0.1 MgSO,7HsO,crams-.-0.2 K.HPO,(anhydrous),grams-0.01Fe(SO,)a(Na),e6H.O,grams.O.aNaCl,grams-._._.,.10.0 Agar,grams-15.0 Distilled water,milliliters-1,000.0 NOTE:R?0.001 ms per liter
9、may be added as an indicator for the presence o!oxygen.19.The indients should be dissolved with gentleheating,The pH should then be adjusted to 7.8 with Naoa If excessive precipitation occu1 the mediumshould be discarded The medium Ja pered and flowed back and f.oJ:th four times to JlllX the inoculu
10、m.One milliliter from this tube fa then transf erred to a second tube and nuxed as be.fore.Contbme thls selial transfer until a dilution al 1 to 10,()QO is reached(6 tubes).After the fnoculum has beent181ls.fet.-red from each tube,the tube should be cooled rapidly to solidify the agar.(Screw caps or
11、 rubber stoppers:should oe used to seal the tubes to prevent dehydration of the medium.)To prevent solidification of a_Bar in the u-ansfer pipettes,warm pipettes(near 45 C)should be uaed for this operation.11.All work is done in duplicate and tubes areincubated at a temperature within 6 0 of the rec
12、orded temperature of the water at the time otsampling.22.All tu s should be held a minimum of 4weeks.The tubes should be examined on the third day and at the end of each week for the a_prance of sulfate-reducing bacteria,as indicated ey intense black colonies.23.The presence of sulfate-reducing bact
13、eria isco11Sidered to represent a potential problem.The extent of.the problem will depend upon additional evidence,such as black water or an increased hydrogen sulfide coo.tent of the injection water.M.A suitable field test for the culwre ot sulfatereducin_g_ bacteria is presented as Appendix A,Sect
14、ion A-Il,Par.A7-A1.SECTION II EVALUATION OF CHEMICALS FOR CONTROL OF MICROBIAL GROWTH Scope 25.These procednres are intended to be used toe-valuate chemicals as anti-microbial control agents in watel-fnjectlon systema where microbial g_rowtb js a-problem,tgtd where it is believed chemical control ex
15、pected to yield a desirable end 41l,These methods are destgned to be Ul3ed for screening purposes in the laboratory.Application to sf:flc mfcrooial-control problems may require modlf1cation if.these techniques(see Appendb:A).Pnrlty of Reagente27.Unless otherwise indicated all chemical rea,.ent:8 shall m.t.Ame1ican Chemical Society speciff.agal and yeast