ML20150D449

New England Coalition on Nuclear Pollution (Necnp) Brief in Support of Appeal of Board Dismissal of Necnp Contention Iv.* Board 880217,0318 & 0401 Decisions Should Be Reversed. W/Certificate of Svc

ML20150D449
Person / Time
Site:	Seabrook
Issue date:	07/01/1988
From:	Ferster A HARMON & WEISS, NEW ENGLAND COALITION ON NUCLEAR POLLUTION
To:	NRC ATOMIC SAFETY & LICENSING APPEAL PANEL (ASLAP)
References
CON-#388-6678 OL-1, NUDOCS 8807140007
Download: ML20150D449 (69)

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{{#Wiki_filter:J ( ccteiu tbHPC UNITED STATES NUCLEAR REGULATORY COMMISSION '88 JL 11 A8 :40 BEFORE THE ATOMIC SAFETY AND LICENSING APPEAL BOARD f OFF!CE < ' biu a +^"I ) DOCKET % 4 'iE"VIC In the Matter of ) BRAhCn ) Public Service Company of ) 50-443 OL-1 t/if 4 04 *j New Hampshire, et al. ) Docket No. ) (Seabrook Station, Units 1 & 2) ) ONSITE EMERGENCY ) PLANNING & TECHNICAL ) ISSUES ) NEW ENGLAND COALITION ON NUCLEAR POLLUTION'S BRIEF IN SUPPORT OF ITS APPEAL OF THE LICENSING BOARD'S DISMISSAL OF NECNP CONTENTION IV t Andrea Ferster Harmon & Weiss 2001 "S" Street N.W. Suite 430 Washington, D.C. 20009 (202) 328-3500 July 1, 1988 o 8807140007 880701 gDR ADOCK 05000443 PDR Dse ..m,, ,,i,

__ _ ___--_-__ __ _ TABLE OF CONTENTS TABLE 13F AUTHORITIES......................................... I. INTRODUCTION.............................................. 1 II. BACKGROUND................................................ 2 III. THE LITERAL LANGUAGE OF CCNTENTION IV ENCOMPASSES THE. ISSUE OF "MICROBIOLOGICALLY INDUCED CORROSION" AND OTHER DETRIMEN-TAL EFFECTS OF "FOULING" OF COOLING SYSTEMS............... 7 IV. THE LICENSING BOARD ACTED ARBITRARILY AND CAPRICIOUSLY IN RESTRICTING THE SCOPE OF NECNP CONTENTION IV............. 11 A. The Licensing Board Applied an Incorrect Legal Standard' in Determining the Scope of NECNP Contention IV..... 11 B. The Licensing Board Erred in Ruling Contrary to the Uncontroverted Expert Affidavit and Scientific Studies Provided by NECNP................................... 14 V. THE LICENSING BOARD ERRED IN DISALLOWING NECNP'S DISCOVERY REQUEST SERVED PRIOR TO THE TIME DISCOVERY CLOSED........ 17 VI. THE LICENSING DOARD ERRED IN DISALLOWING DISCOVERY INTO CIR-CULATING WATER SYSTEMS THAT WERE NOT "COOLING SYSTEMS"... 19 VII. C O N C LUS I ON............................................... 2 1 EXHIBITS A - G - - + ev +-

. _ _ _ _ _ _ _ - _ _ TABLE OF AUTHORITIES Cases Page Culler v. Commissioner of Internal Revenue, 237 F.2d 611 (8th C i r. 19 5 6 ).................................................... 1 6 Pike v. U.S., 340 F.2d 487 (9th Cir. 1974)..........*.......... 11 Administrative Decisions Carolina Power and Licht Co. (Shearon Harris Nuclear Power Plant), ALAB-852, 24 NRC 532 (1986).................... 9-10, 11 Cincinnati Gas and Electric Co. (William H. Zimmer Nuclear Sta-tion), 12 NRC 231 (1980)...................................... 19 Pacific Gas and Electric Co. (Stanislaus Nuclear Project, Unit 1, LBP-78-20, 7 NRC 1038 (1978).................................. 20 Philadelohia Electric Co. (Peach Bottom Atomic Power Station, Units 2 and 3), ALAB-216, 8 AEC 13 (1974)................. 10, 12 Philadelphia Electric Co. (Limerick Generating Station, Units 1 and 2), 22 NRC 681 (1985)..................................... 12 Texas Utilities Co. (Comanche Peak Steam Electric Station, Unit 1, ALAB-868, 25 NRC 912 (1987)........................... 10,, 11 Iss.an Utilities Generatina Co. (Comanche Peak Steam Electric Sta-tion, Units 1 and 2), LPB-81-25, 14 NRC 241 (1981)............ 20 l Reculations l 10 C.F.R. 5 2.714(b).................................. 10, 11, 14 10 C.F.R. 5 2. 7 4 0 ( b ) ( 1 )....................................... 21 Miscellaneous

Bryers, J.D.,

Characklis, W.G.,

Zelver, N.,

and Nimmons, M.G., "Microbial Film Development and Associated Energy Losses," at 12.14-1, Paper No. 12-15 presented at the Proc. 6th OTEC Confer-ence, "Ocean Thermal Energy for the '80's," Washington, D.C., June 19-20, 1979............................................ 8, 9 Characklis, W.G., "Bioengineering Report -- Fouling Biofilm l Development: A Process Analysis." Biotechnoloav and Bioencineer-ing, Vol. XIII (John Wiley & Sons, Inc. 1980).................. 9 1 1

-lii-Neitzel, at al, "Improving the Reliability of Open-Cycle Water Systems: An Evaluation of Biofouling Surveillance and Control Techniques for Use at Nuclear Power Plants," NUREG/CR-4724, Vol. 1 (1986).................................................... 3, 4 Norman, G.,~Characklis, W.G., and Bryers, J.D.,*"Control of Microbial Fouling in Circular Tubes with Chlorine," 18 Develgn-ment in Industrial Microbioloav (1977)........................ 13 Stein, Mitchell, and Mezines, Administrative Law, S 28.06 (Mathew-Bender, 1987)......................................... 16 r

July 1, l'908 UNITED STATES NUCLEAR REGULATORY COMMISSION BEFORE THE ATOMIC SAFETY AND LICENSING APPEAL BOARD ) In the Matter of ) ) Public Service. Company of ) New Hampshire, et al. ) Docket No. 50-443 OL-1 ) (Seabrook Station, Units 1 & 2) ) ONSITE EMERGENCY ) PLANNING & TECHNICAL ) ISSUES ) NEW ENGLAND COALITION ON. NUCLEAR POLLUTION'S BRIEF IN SUPPORT OF ITS APPEAL OF THE. LICENSING BOARD'S DISMISSAL OF NECNP CONTENTION IV I. INTRODUCTION The instant appeal concerns the Licensing Board's March 18, 1 1988, ruling which limited the scope of NECNP Contention IV to only one of the adverse affects of the accumulation of aquatic organisms,in cooling systems, namely, the accumulation of macro-organisms resulting in blockage and constric. ion of coolant flow. As a result of this ruling, as well as other Licensing Board .v rulings constricting the time and type of allowable discovery for this contention, NECNP was precluded from inquiring into, or litigating, the adequacy of Applicants' program to monitor and 1 NECNP Contention IV reads as follows: The Applicent must esta.blish a survellance and maintenance program for the preven-tion of the accumulation of mollusks, other aquatic organisms, and debris in cooling systems in order to satisfy the require-ments of GDC 4, 30, 32, 33, 34, 35, 36, 38, and 39, which require l the maintenance and inspection of reactor cooling systems. The i design, construction and proposed operation of Seabrock fail to l satisfy these requirements. l l

s control microbiologically induced corrosion, a form of biofouling caused by the accumulation of microbiological organisms in cool-ing systems. II. BAQKGROUND On March 25, 1987, the Licensing Board issued a Partial Ini-tial Decision ("PID") Which authorizes Public Service Co. of New Hampshire ("Applicants") to operate the Seabrook nuclear power plant at power levels up to and includ ing 5% of rated power.2 NECNP appealed that decision on the merits, arguing, inter alia, that the Licensing Board had wrongly den 19d NECNP Contention IV. On October 1, 1987, the Atomic Safety and Licensing Appeal Board (the "Appeal Board") issued ALAB-875, reversing and remanding in part the March 25, 1987 Licensing Board dccision authorizing a low power license for Seabrook. The Appeal Board ordered, inter alia, that the Licensing Board admit two of NECNP's contention concerning protection against steam generator tube ruptures (NECNP Contention I.V.) and potential degrading of the plant's heat removal capability due to build-up of biological organisms (NECNP Contention IV), and begin the litigation process for these improperly rejected contentions.3 Discovery upon the remanded 2 Public Service Co. of New Hampshire, et al. (Seabrook Station, Units 1 and 2), LBP-87-10, 25 NRC 177 (1987). Hereinafter, all administrative decisiors in the Seabrook proceeding will be cited ( only by number and date. The agency's citation system denotes l decisions of the Licensing Board Panel as "LBP" decisions, Appeal Board decisions as "ALAB," and Commission decisions as "CLI." 3 ALAB-875, slip op, at 13-20.

. contentions began on October 26, 1987, and was to be completed by February 19, 1988.4 In response to NECNP's Second Set of Interrogatories and Request for Production of Documents to Applicants on NECNP Con-tention IV, dated December 23, 1987, Applicants objected to NECNP's questions concerning the adequacy of Applicants' program to detect and control "microbiologically induced corrosion," on the ground that the literal language of NECNP Contention IV did not specifically identify "microbiologically induced corrosion" as an issue.5 NECNP then filed a motion to compel Applicants to respond to these questions, in which NECNP presented proof, in the form of NRC-sponsored studies,6 demonstrating that the literal language used in NECNP Contention IV and bases placed Applicants on notice that the issue of "microbiologically induced i l l t l 4 Discovery was extended by the Licensing Board Order of Decem-ber 2, 1987, to take into account the additional obligations placed on counsel as a result of the Appeal Board rulings on low power operation. 5 "Applicants' Responses to NECNP's Second Set of Inter-rogatories and Request for Production of Documents to Applicants on NECNP Contention IV," filed January 14, 1988, at 2. 6 Neitzel, et al, "Improving the Reliability of Open-Cycle Water Systems: An Evaluation of Biofouling Surveillance and Control Techniques for Use at Nuclear Power Plants," NUREG/CR-4724, Vol. 1 (1986).

-4 corrosion" was encompassed by the contention.7 By Order dated February 17, 1988, the Licensing Board denied NECNP's motion to compel, and ruled, inAgr alia, that the issue of "microbiologi-cally induced corrosion" ("MIC") is not within the scope of NECNP Contention IV. The Licensing Board reasoned that "NUREG/CR-4724 was issued some four years after Contention IV was proposed," and that'NECNP cannot "expand the scope of the contention by reliance upon a document that did not exist at the time contention IV was submitted.n8 NECNP then sought and was granted leave to file a motion for reconsideration of the Licensing Board's February 17, 1988 Order, which motion was filed by NECNP on March 1, 1988. In support of its motion, NECNP presented an expert affidavit from Dr. James Bryers, who testified that the scientific meaning of the literal terms of Contention IV encompassed the issue of microbiological 1y l l 7 "NECNP's Motion to Com,,el Applicants to Respond to NECNP's Second Set of Interrogatories and Request for Production of Docu-ments on NECNP Contention IV," dated January 25, 1988. NECNP also moved to compel Applicants to respond to questions seeking l information concerning possible biofouling and corrosion in "cir-culating water systems" at the Seabrook plant. Applicants had refused to provide this information with respect to all circulat-ing water systems which Applicants claim are not "cooling sys-tems" based on their view that "Issues concerning circulating water systems generally are outside the scope of contention IV." "Applicants' Responses to NECNP's Second Set of Interrogatories and Request for Production of Documents to Applicants on NECNP Contention IV," filed January 14, 1988, at 13, 28. The Licensinj l Board did not address this argument, since it viewed its resolu-tion of the MIC issue as dispositive. ASLB Memorandum and Order (unpublished], dated February 17, 1988, at 3 n. 2. l 8 Memorandum and Order of February 17, 1988 Denying NECNP's Motion to Compel, (unpublished) at 6-7. i

L induced corrosion.9 In addition, NECNP submitted scientific studies contemporaneous to the admission of contention IV demonstrating-that, in 1982, microbiologically induced corrosion was recognized as one of the detrimental effects of biofouling of nuclear power plants.10 Both the Applicants and the NRC Staff filed responses opposing NECNP's motion, which urged the Licens-ing Board to disregard the expert affidavit of Dr. Bryers on the ground that it was "unpersuasive;" however, they failed to pro-vide anY expert opinion or studies of their own controverting the voluminous material provided by NECNP.11 The Licensing Board then denied NECNP's motion for reconsideration, again on the grounds that "the opinion of Dr. Bryer (sic) and the appended scier.tific studies cannot serve to establish that, in preparing the contention in 1982, the drafter intended to encompass MIC within the scope of the contention.n12 9 NECNP's affiant, Dr. James Bryers, is a professor in the Cen-ter for Biochemical Engineering at Duke _ University, and is the author of over thirty published articles in scientific journals and treatises on the subject of microbial fouling and its effects in engineered systems, including nuclear power plant heat-exchange systems. Dr. Bryers' affidavit and curriculum vitae are l attached in support of the instant appeal, as Exhibits A and D. 10 copies of the relevant portions of these studies are attached hereto as Exhibits C through G. 11 "Applicant's Response to NECNP's Motion for Reconsideration of the Board's Order Denying NECNP's Motion to Compel," dated l March 14, 1988, at 3; "NRC Staff Reponse to NECNP Motion for Reconsideration of the Board's Denial of NECNP's Motion to Com-l pel," dated March 11, 1988, at 5 n.3. I 12 Memorandum and Orde).- (unpublished), dated March 18, 1988, at 3. l.

-.____ ______, On February 19, 1988, NECNP filed a request for entry upon land for inspection and the taking of notes, samples and photog-raphs at Seabrook in the areas containing piping and heat-exchangers that are. susceptible to biofouling or micrebiologi-cally induced corrosion, and in laboratories where testing for biofouling or microbiologically induced corrosion is performed. The Licensing Board denied NECNP's request on the ground that it was untimely, and because it concerned matters not within the scope of NECNP Contention IV.13 On March 22, 1988, NECNP filed a motion to compel Applicants to respond to NECNP's Third Set of Interrogatories and Request for production of documents to Applicants on NECNP Contention IV, which were served on February 19, 1988. The Licensing Board denied this motion, again, on the ground that the issue of micro-biologically induced corrosion was not encompassed within the scope of NECNP Contention IV.14 By letter dated April 22, 1988, NECNF notified the Licensing Board and the parties that it did not choose to litigate the Con-tention IV due to the Licensing Board's restrictive rulings which precluded NECNP from litigating the adequacy of Applicants' pro-gram for controlling microbiologically induced corrosion. NECNP further stated that it intended to appeal the Licensing Board's 4 13 ASLB Memorandum and Order (unpublished), dated March 18, 1988, at 4-5. 14 ASLB Order (unpublished), dated April 1, 1968.

+ \\ rulings on the scope of NECNP Contention IV at the appropriate time. On May 12, 1988, in reaction to this letter, the Licensing Boarddismissed NECNP Contention IV as "abandoned." NECNP filed a notice of appeal on June 1, 1988, along with a mdtion for leave to file the notice of appeal out of time. The Appeal Board, in ALAB-894, granted NECNP's motion.15 III. THE LITERAL LANGUAGE OF' CONTENTION IV ENCOMPASSES THE ISSUE OF "MICROBIOLOGICALLY INDUCED CORROSION" AND OTHER DETRIMENTAL EFFECTS OF "FOULING" OF COOLING SYSTEMS, It is important, at the outset, to understand the scientific definitions of the phenomenon of "biofouling" and "microbiologi-cally induced corrosion," and the scientific meaning of the terms employed in NECNP Contention IV. NECHP Contention IV asserts simply that "the Applicant must establish a surveillance and maintenance program for the prevention of the accumulation of mollusks, other aquatic organisms, and debris in cooling sys-tems..." This contention does not specifically identify the problem as either "fouling" or "biofouling," nor does it identify any of the detrimental effect's of this process. However, this language in fact broadly identifies the process commonly known as "fouling," which has a number of detrimental effects, including blockage, constriction and/or mechanical deterioration of the 15 A LAB-89 4.

3 v . operating characteristics of valves and pumps, fluid resistance, and-corrosio,n.16 The Licensing Board determined that NECNP Contention IV was. limited to only one particular detrimental effect of fouling, that:of blockage of piping in heat exchange systems, which is generally caused by the build-up of macro-organisms (such as clams and mussels) and debris on.the inside of piping,'resulting + in'the degradation of heat removal capability of cooling water systems. In reaching this conclusion, the Licensing Board appropriately relied on the literal language of the contention, which referred to "the accumulation of mollusks, other aquatic organisms, and debris."17 However, the Licensing Board erred in' reaching the conclusion the word "accumulation" was intended to refer only to the detrimental effect of blockage caused by_ foul-ing.- Rather, the term "accumulation" clearly refers to the pro-cess.of fouling itself, which is the accumulation of organisms 16 Egg Bryers' Affadivit, Exhibit A, at 7; gag also Bryers, J.D., Characklis, W.G.,

Zelver, N.,

and Nimmons, M.G., "Microbial Film Development and Associated Energy Losses," at 12.14-1, Paper No. 12-15 presented at the Proc. 6th OTEC Conference, "Ocean Thermal Energy for the '80's," Washington, D.C., June 19-20, \\ 1979, which defines "fouling" as follows: The term fouling refers to the formation of inorganic and/or organic deposits on'aurfaces. In cooling systems, these deposits form on condenser tube walls increasing fluid fric-tional resistance, accelerating corrosion and impairing heat transfer. An excerpt of this study is attached hereto as Exhibit C. i l 17 ASLB Memorandum and Order, dated February 17, 1988, at 5 (emphasis in original).

X U y T (both macro and micro) and debris on pipes. The accumulation of macro-organisms can cause blockage, and the accumulation of biofilms on heat-exchange systems can ultimately interact with ~ bacteria to cause the chenomenon of microbiologically induced corrosion.18 Thus, the use of the word "accumulation" in the Contention reinforces a bread, rather than limited construction of Contention IV. I* is well settled that a party is bound by the literal terms of its own contention. Carolina Power and Licht Co. (Shearon Harris Nuclear Power Plant), ALAB-852, 24 NRC 532, 545 18 Perhaps the clearest explanation of the process, and dif-ferent types of fouling, appears in a 1981 article by W.G.Characklis, entitled "Bioengineering Report -- Fouling Biofilm Development: A Process Analysis:" The term fouling refers to the formation of inorganic and/or organic deposits on surfaces. These deposits can impede the flow of heat ~across the surface, increase the fluid frictional resistance at the surface, and increase the rate of corrosion at the surface. In any case energy losses result. Several types of fouling and their combinations may occur in heat exchangers: 1) crystalline or precipitation fouling, 2) corrosion fouling, 3) particulate fouling, 4) chemical reaction fouling, and 5) biological fouling. Biological fouling results from a) development of a biofilm consisting of microorganisms and their products (microbial fouling), b) deposition and growth of macroorganisms :such as barnacles (macrobial fouling), and c) assorted detritus. Bio _technoloav and Bioencineerina, Vol. XIII, pp. 1923-1960 (John Wiley & Sons, Inc. 1980), excerpt attached as Exhibit D. See also Bryers, J.D., Characklis, W.G.,

Zelver, N.,

and Nimmons, M.G., "Microbial Film Development and Associated Energy Losses," at 12.14-1, Paper No. 12-15 presented at the Proc. 6th OTEC Con-ference, "Ocean Thermal Energy for the '80's," Washington, D.C., June 19-20, 1979, excerpt attached hereto as Exhibit C; Bryers Affidavit, at 8.

s o v (1986). Here, the literal terms of the contention broadly identify the process of fouling, which_has a number of detrimen-0( tal effects, including microbiological 1y induced corrosion. Accordingly, microbiologically induced corrosion is within the scope of NECNP Contention IV. The Licensing Board, in part, apparently based its decision on the fact NECNP Contention IV did not specifically identify the issues of "biofouling" or "microbiologically induced corrosion." However, a contention need only be specific enough to put other parties on notice so that they will know what to defend against or oppose, and to assure that the proposed issues are proper for adjudication. Philadelphia Elqctric Co. (Peach Bottom Atomic Power Station, Units 2 and 3), ALAB-216, 8 AEC 13, 20 (1974). The fact that the contention fails.to identify specifically the issues encompassed by the contention by their technical names cannot preclude litigation of those issues, as that would estab-lish "secretive and complex technicalities" not intended by the basis and specificity requirements of 10 C.F.R. S 2.714(b). Id. l In case of doubt, Applicants "may fill any gaps in their knowl-l edge of the intervenors' case through discovery against inter-i venors." Texas Utilities Co. (Comanche Peak Steam Electric Sta-l tion, Unit 1, ALAB-868, 25 NRC 912, 933 (1987).19 l I l 19 We note that Applicants here chose not to undertake any dis-covery against NECNP; in any event, however, they were quickly l put on notice through NECNP's discovery that NECNP regarded microbiologically induced corrosion as within the scope of its contention.

4 s e v i Here, NECNP Contention IV, by broadly identifying the pro-cess of fouling of nuclear power plant heat-exchange systems, clearly put Applicants en notice that the contention encompassed all the detrimental effects of this process, including micro-biologically induced corrosion. Thus, the issue of microbiologi-cally induced corrcsion was identified with sufficient specificity to satisfy the pleading requirements of 10 C.F.R. 5 2.714(b).20 IV. THE LICENSING BOARD ACTED ARBITRARILY AND CAPRICIOUSLY IN RESTRICTING THE SCOPE OF NECNP CONTENTION IV. A. The Licensing Board Applied an Incorrect Legal Standard in Determining the Scope of NECNP Contention IV. As noted above, the scope of a contention is defined by the literal terms of the contention. Carolina Power and Light Co. (Shearon Harris Nuclear Power Plant), ALAB-852, 24 NRC E32, 545 (1986). However, the Licensing Board wholly disregarded the voluminous expert and scientific evidence presented by NECNP that the literal language of contention IV encompassed the issue of microbiologically induced corrosion. Instead, the Licensing 20 While microbiologically induced corrosion is not specifically discussad in the bases for the contention, the bases for a con-tention cannot be relied on to alter the contention's actual lan-l guage. Texas Utilties Co. (Comanche Peak Steam Electric Station, Unit 1), 25 NRC 912, 932 n. 83 (1987). And, es noted above, the l contention's literal language was sufficient to identify the issue of microbiological 1y induced corrosion. Likewise, the use of the word "blockage" in the caption of NECNP Contention IV can-not be construed as limiting the scope of the contention to only that detrimental effect of fouling, to the exclusion of others. l Like the interpretion of statutes, titles or captions cannot be i j used to alter or vary the plain meaning of provisions, gen e.a1 l Pike v. U.S., 340 F.2d 487 (9th Cir. 1974).

i ,L Board created an entirely novel standard which seeks to determine NECNP's "intent" when its contention was formulated. Applying this new standard, the Licensing Board determined that NECNP did .b not "intend" for its contention to encompass the issue of micro-biologically induced corrosion in 1982, when the contention was first formulated. y This subjective standard for determining the scope of a con-tention is without any support in past NRC precedent.21

Further, it is flatly contradicted by the numerous cases setting forth an i

obiectivq test for determining the scope of a contention, namely, that the literal language employed by the contention controls. Egg Philadelchia Electric Co. (Limerick Generating Station, Units 1 and 2), 22 NRC 681, 709 (1985), in which the Appeal Board held the intervenors to the literal terms of its contention, despite their assertion that the "sought to litigate something else." More importantly, the subjective standard applied by the Licensing Board runs contrary to the principle purpose of 10 C.F.R. 5 2.714(b), which is to provide notice to Applicants of so that they will know what to defend against or oppose. Philadel-phia Electric Co.. (Peach Bottom Atomic Power Station, Units 2 and 3), ALAB-216, 8 AEC 13, 20 (1974). Indeed, as the Limerick case 21 Ironically, the Licensing Board was forced to confine its search for "NECNP's then (1982) intent" to the language and basis of the contention itself, since this contention was wrongly rejected by the Board at the outset, thereby precluding any opportunity to develop this contentior. As noted above, the literal language of the contention itself encompasses t*s issue of microbiologically induced corrosion.

,e e r lt / - 4 cited above recognizes, it'would be fundamentally unfair to / . Applicants if subjective intent could be used to guide litigation c '\\ of contentions.and to protect Applicants from surprises. Indeed, NECNP presented evidence that microbiologically ~

• induced corrosion was recognized as one of the detrimental effects of biofouling as early as 1977, five years prior to the formulation of NECNP Contention IV.22 Thus, the literal' language of the contention was sufficient to provide adequate notice to Applicants that microbiologically induced corrosion was one of the issues to be litigated within the scope of NECNP Contention IV in 1982, when the contention was drafted, as well as in 1987 I

1 -- the more' relevant time period -- when the contention was actually admitted and litigation begun. Despite the fact that this evidence was uncontroverted, the Licensing Board disregarded this evidence, again on the premise that these studies did not l i indicate what NECNP "intended" in preparing the contention 1982.23 Clearly, the focus snould not be on what NECNP l "intended" but on whether Applicants objectively had adequate notice. We submit that the literal language of the contention provided this notice. l I l l l 22 Bryers' Affidavit, at 9; Egg also Norman, G., Characklis, l W.G., and Bryers, J.D., "Control of Microbial Fouling in Circular Tubes with Chlorine," 18 Develcoment in Industrial Microbioloav, pp. 581-590 (1977), excerpt attached as Exhibit E. 23 ASLB Memorandum and Order (unpublished), dated March 18, [ 1988, at 3. l l t #

F x Moreover, the facts here show that NECNP did intend that microbiologically induced corrosion be encompassed within the scope of its contention. This intent is apparent in the conten-tion's literal language, which broadly identified the issue of fouling, and in NECNP discovery requests, in which NECNP con-sistently sought to acquire information about the Applicants' 3 program to control microbiologically induced corrosion. The Licensing Board erred in basing its finding of NECNP's "1982 intent" on the very general information provided to satisfy the basis and specificit) requirements of 10 C.F.R. S 2.714(b), and in ignoring the more persuasive evidence of NECNP's intent con-sistently avidenced in "ECNP's discovery requests and pleadings, which were filed as saon as NECNP was permitted to litigate the contention. Again, to confine the search for NECNP's "intent" to only those documents filed in 1982, and to ignore the clear evi-dence of NECNP's intent contained in its discovery requests, would unfairly penalize NECNP for the Licensing Board's action in wrongly rejecting the contention at the outset. B. The Licensing Board Erred in Ruling Contrary to the Uncontroverted Expert Affidavit and Scientific Studies Provided by NECNP. In addition to applying an incorrect legal standard, the Licensing Board erred in disregarding the expert affidavit and scientific studies presented by NECNP. NECNP presented an expert affidavit from Dr. James Bryers, one of the nation's foremost experts on the issue of microbiologically induced corrosion and fouling of heat-exchange systems, who stated that the literal

n s - t , ~ language of the contention encompassed the issue of microbiologi-cally induced corrosion,24 and that the problem of corrosion in engineered systems caused by the interaction between micro-organisms and biofilms on pipe-liquid surfaces was recognized by the scientific community as early as 1977.25 Neither the Applicants nor the Staff offered any controverting affidavits or studies. Rather, Applicants rested merely on the bald assertions of counsel that Dr. Bryers' testimony is "unpersuasive;n26 and a the NRC Staff merely stated,,aga,in without submitting any con-troverting evidence or expert opinion, that Dr. Bryers' opinion "is entitled to little, if any weight.n27 Despite the absence of ADY controverting evidence, the Licensing Board disregarded Dr. Bryers' testimony. The Licensing Board's ruling was based on its view that Dr. Bryers' statements, by necessity made six years after the contention was formulated, "cannot serve to establish that, in preparing the contention in 1982, the drafter intended 24 Bryers' Affidavit, at 5. 25 Bryers' Affidavit, at 9; See also Norman, G., Characklic, W.G., and Bryers, J.D., "Control of Microbial Fouling in Circular Tubes with Chlorine," 18 Development in Industrial Microbioloav, pp. 581-590 (1977), excerpt attached as Exhibit E. 26 "Applicant's Response to NECNP's Motion for Reconsideration of the Board's Order Denying NECNP's Motion to Compel," dated March 14, 1988, at 3. 27 "NRC Staff Reponse to NECNP Motion for Reconsideration of the Board's Denial of NECNP's Motion to Compel," dated March 11, 1988, at 5 n. 3.

X c 1 to encompass MIC within the scope of the contention.n28 Where expert opinion evidence is submitted by only one side, as is the case here, an agency may disregard it only under three circumstances: where the agency possesses th'e expertise to sub-stitute its judgment in the place of the experts'; where their is contrary evidence already in tha record; and where the expert's testimony has minimum credibility. Stein, Mitchell, and Mezines, Administrative Law, S 28.06 (Mathew-Bender, 1987). None'of these circumstances is present here. First, the issue at hand involves the interpretation of technical, scientific termu used in the field of microbiology and biochemical engineering, wh!ch is not an area in which the Com-mission possesses expertise. Where the testimony of a witness is in an area in which the agency lacks knowledge or technical skill, it may not arbitrarily substitute its judgment for that of an expert witness. Culler v. Commissioner of Internal Revenue, 237 F.2d 611, 616 (8th Cir. 1956). Second, there was no contrary evidence in the record, other j than the unsupported "lay" opinion of Applicants' and the Staff's l legal counsel. Finally, as noted above, Dr. Bryers is one of the l I l l l 28 ASLB Memorandum and Order (unpublished), dated March 18, 1988, at 3. This reasoning is particularly ironic, in light of the fact that Dr. Bryers' "Dost hgg" interpretation of this Con-tention is necessitated by the fact that the contention was wrongfully dismissed at an earlier stage in this proceeding. To l . disallow expert opinion as to the scientific meaning of the plain langauge of the Contention because it could not, due to an error l not of NECNP's making, be made contemporaneously, is blatantly unfair and prejudicial to NECNP. 1

e - 17'- country's foremost experts on the subject of the effects of biological fouling on engineered safety systems. His opinion as to the meaning and scope of the plain language of NECNP Conten-tion IV is. clearly entitled to great weight.29 Accordingly, the Licensing Board abused its discretion in ruling contrary to the uncontroverted and entirely tredible evidence presented by NECNP that microbiological 1y induced corrosion was within the scope of NECNP Contention IV. V. THE LICENSING BOARD ERRED IN DISALLOWING NECNP'S< DISCOVERY REOUEST SERVED PRIOR TO THE TIME' DISCOVERY CLOSED. The Licensing Board denied NECNP's Motion for Leave to Enter Applicants' Land, filed on February 19, 1988, and its alternative motion to extend the dead)!ne for discovery,30 on two grounds: first, that the motion sought discovery on matters not within the scope of NECNP Contention IV; and second, that the motion was untimely.31 This ruling was in error on both counts. l l 29 While Dr. Bryers' opinion as to "the scope of NECNP Conten-tion IV" is, admittedly, the ultimate issue of this case, Dr. Bryers' expert opinion of the technical, scientific meaning of the terms used in the contention, and his expert opinion as to the range of detrimental effects that are caused by the process referred to in the contention, are entirely appropriate and admissible. 30 "NECNP's Reply to Applicants' Response to NECNP's Request for Entry Upon Land," dated March 3, 1988, at 4. 31 ASLB Order (unpublished), dated March 18, 1988, at 4-5. According to the Licensing Board, its December 2, 1988 Scheduling l Order, which provided that "Applicants,' NECNP and the Staff shall... complete discovery by February 19, 1988," meant that all responses to discovery must be received by February 19, 1988. l n

w 1 , o First, as noted above, microbiologically induced corrosion and biofouling are within.the scope of NECNP Contention IV. Therefore, NECNP's motion was clear y "relevant to the subject ? ~ matter involved in the proceeding...(and) appears reasonably cal-culated to lead to the discovery of admissible evidence." 10 C.F.R. 9 2.740(b) (1). Second, the language used in the Licensing Board's December 2, 1987 Scheduling Order implied that February 19, 1988 was the last date on which reauests for discovery may be served. The Board's Scheduling Order did not provide a separate deadline by which discovery requests must be served and received. Past NRC practice in the Seabrook proceeding has consintently been that the date by which discovery is to be closed has meant the date on-which the last discovery request must be filed.32 Rather; where a Licensing Board han intended to impose on parties a specific deadline by which the last discovery response must be received, and depositions must be taken, it has done so explicitly.33 NECNP relied in good faith on the Board's previous practice 1 of providing explicit guidelines and deadlines in such instances where it intended for the deadline for service of discovery requests to be different and earlier from deadline for completion i 32 "Memorandum & Order - Establishing Hearing Schedule on Offsite Issues Raised By NHRERP," ASLBP No. 82-471-02-OL, dated December 4, 1986; "Memorandum and Order," ASLPBP No. 82-471-02 OL, dated September 13, 1982. 33 "Memorandum and Order," ALBP No. 82-471-02-OL, dated July 25, 1986, at 11-12.

s or closure of discovery. Given the serious consequences in terms of NECNP's ability to effectively litigate the important, remanded safety issue of NECNP Contention IV, and the reasonable-ness of NECNP's reliance on past practice regarding discovery scheduling, the Board should have allowed NECNP's motion, or granted NECNP's request in the alternative for an extension of the discovery deadline. Sha Cincinnati Gas and Electric Co u (William H. Zimmer Nuclear Station), 12 NRC 231, 232 n.1 (1980) ) (ASLB considered untimely filed response because reluctant to take position which might preclude litigation of safety or environmental issues without giving every party an opportunity to 1 be heard). VI. THE LICENSING BOARD ERRED IN DISALLOWING DISCOVERY INTO CIR-CULATING WATER SYSTEMS THAT WERE NOT "COOLING SYSTEMS. " The Licensing Board also erred in denying NECNP's motion to compel Applicants to respond to questions seeking information concerning possible biofouling and corrosion in all "circulating water systems" at the Seabrook plant, on the ground that NECNP Contention IV only concerned "cooling systems."34 These dis-covery requests were entirely permissible. It is well established that "In modern administrative and l legal practice, pretrial discovery is liberally granted to enable the parties to ascertain the facts in complex litigation, refine 34 "NECNP's Motion to Compel Applicants to Respond to NECNP's Second Set of Interrogatories and Request for Production of Docu-ments on NECNP Contention IV," dated January 25, 1988, at 4-5.

a / ~20-the issues, and, prepare adequately for a nore expeditious hearing or trial." Pacific Gas and Electric Co. (Stanislaus Nuclear Pro-ject, Unit 1, LBP-78-20, 7 NRC 1038, 1040 (1978). In this 4 regard, interrogatories need only have "general relevance, for. discovery purposes, to the matters in controversy in the proceed-ing." Texas Utilities Generatina Co. (Comanche Peak Steam Elec, tric Station, Units 1 and 2), LPB-81-25, 14 NRC 241, 243 (1981). a Here, NECNP sought information about other systems in order to determine the extent to which microbiologically induced corro-sion has occurred and is adequately treated in general at Seabrook. These interrogatories seeking information about other systems are designed to lead to information that ir, relevant to NECMP's Contention IV, which concerns the adequacy of Applicants' surveillance and maintenance program for the prevention of micro-biologically irducc3 corrosion in cooling systems. The presence of corrosion in other circ. dating water systems may well indicate that corrosion may occur in cooling systems. If corrosion has occurred in other circulating water systems, it is necessary to determine whether Applicants have a program designed to prevent or control corrosion in these systems, obviously, if Applicants' responses showed that these programs are the same as the programs used to prevent or control corrosion in cooling systems, this may be admissible evidence that such programs are also not adequate to treat or control corrosion in cooling systems. Accordingly, these interrogatories were clearly "relevant to the subject mat-ter involved in the proceeding...[or which] appears reasonably

F. calculate'dcho lead to the discovery of admissible evidence." 10 C.F.R. $ 2.740(b) (1). VII. CONCLUSION For the foregoing reasons, the Licensing Board's erred in ruling that microbiologically induced corrosion was not wittin the' scope of NECNP Contention IV, and by restricting the. time and type of allowable discovery under this contention. Therefore, the Licensing Board's decisions of February 17, 1988, March 18, 1988, and April 1, 1988, should be reversed. Respectfully submitted _, p - T ???-r f f&l*-. Andrea Ferster HARMON & WEISS 2001 "S" Street N.W. Suite 430 Washington, D.C. 20009 (202) 328-3500 CERTIFICATE OF SERVICE I certify that on July 1, 1988, copies of the foregoing pleading were served by first-class mail, or as otherwise indi-cated, on all parties listed on the attached service list. W f:%gg fM /.: e Andrea forster l l l l

t en ! o ~ / y/ / SEABROOK SERVICE LIST - ONSITE APPEAL BQARD 3 '] Sheldon J. Wolfe, Chairman 19th Floor Ropes & Gray - -4 'JUL 1119885 9 U.S. NRC Boston, MA 02108 225 Franklin Street 6 LSandraGavutis. 4 RFD ifBoil154C E >h E5s"t'Kessingth H 0382 Washington, D.C. 20555 Boston,MA 02110 Stanley W. Knowles Dr. Jerry Hart)our Board of Selectmen Robert A. Backus, Esq. /7yyppQ\\ U.S. NRC P.O. Box 710 Backus,Meyer & Sdomon 'Charlesh raham, Esq. Washington, D.C.20555 North Hampton, NH 03826 111 LowellStreet McKay, Murphy and Graham Manchester,NH 03105 100 Main Street "Dr. Emmeth Luebke J.P. Nadeau Amesbury,MA 01913 5500 Friendship Blvd. Town of Rye Gregory A. Berry, Esq. Apartment #1923N 155 Washington Road Office of den. Counsel Chesy Chase, MD 20815 Rye, NH 03870 U.S. NRC Washington, D.C. 20555 " Hand Delivery "Alan S. Rosenthal, Richard E. Sullhan Mayor

• Overnight Mail Chairman City Hall R. Scott Hill.Whilton U.S. NRC Newburyport, MA 01950 Lagoulis, Clark, Hill Whilton Washington, D.C. 20555

& McGuire Alfred V. Sargent Chairman 79 State Street "Howard A.Wilber Board of Selectmen Newburyport, MA 01950 U.S. NRC Town of Salisbury, MA 01950 Washington, D.C.20555 George Dana Bisbee, Esq. Sen. Gordon Humphrey Geoffrey M. Huntington, Esq. Atomic Safety & Licensing U.S. Senate Office of the Attorney General Board Panel Washington, D.C. 20510 State House Annex U.S. NRC (Atta. Tom Burack) Concord, NH 03301 Washington, D.C. 20555 Selectmen of Northampton Allen Lampert Atomic Safety & Licensing Northampton, NH 03826 Chil Defense Director AppealBoard Panel Town of Brentwood U.S. NRC Sen. Gordon Humphrey. Exeter,NH 03833 Washington, DI;. 20555 1 Eagle Square Suite #507 Richard A. Hampe, Esq. Docketing and Service Concord,NH 03301 Hampc and McNichoin U.S. NRC 35 Pleasant Street j ' Washington, D.C. 20555 MichaelSantosuosso Concord,NH 03301 Chairman Mrs. Anne E. Goodman Board of Selectmen Gary W. Holmes, Esq. Board of Selectmen Jewell Street, RFD #2 Holmes & Ellis 13-15 New Market Rd. South Hampton, NH 03842 47 Winnacunnent Road Durham, NH 03842 Hampton,NH 03842 Judith H. Mizner, Esq. William S. Lord, Selectman Siherglate, Gertner, et al. William Armstrong Town Hall - Friend St 88 Broad Street Civil Defense Director Amesbury,MA 01913 Boston, MA 02110 10 Front Street Exeter, NH 03833 Jane Doughty Rep. Roberta C. Pevear SAPL Drinkwater Road Cahin A.Canney 5 Market Street Hampton Falls, NH 03844 City Manager, City Hall Portsmouth,NH 03S01 126 Daniel Street Phillip Ahrens, Esq. Portsmouth,NH 03S01 l Carol S. Sneider, Esq. Asst. Attorney General l Assistant Attorney General State House, Station #6 Matthew T. Brock, Esq. 1 Ashburton Place Augusta, ME 04333 Shaines & McEachern P.O. Box 3fo

• Thomas G. Dignan, Esq.

Maplewood Avenue R.K. Gad II, Esq. Portsmouth,NH 03801

"7 + i UNITED STATES NULLEAR REGULATORY COMMISSION BEFORE THE ATOMIC SAFETY AND LICENSING BOARD ) In the Matter of ) ) Public Service Company of ) New Hampshire, et al. ) Docket No. 50-443 OL-1 ) (Seabrook Station, Units 1 & 2) ) ONSITE EMERGENCY ) PLANNING & TECHNICAL ) ISSUES ) AFFIDAVIT OF DR. JAMES BRYERS I, James Bryers, being on oath, depose and say as.follows: 1. I am a Professor in the Center for Biochemical Engineering at Duke University. My curriculum vitae, which describes my academic and professional experience, publications, and research, is attached hereto as Exhibit B. 2. My area of expertise in the field of chemical engineering is in the physical, chemical and biological processes a governing biofilm formation, and the detrimental effects of these biofilms in engineered systems, including nuclear power plant j heat-exchangers. 4 3. I have reviewed the text and bases of Contention IV, "Blockage of Coolant Flow to Safety-Related Systems and Com-1 ponents by Buildup of Biological Organisms," sponsored by New England Coalition on Nuclear Pollution ("NECNP") in the above-captioned proceeding. It is my opinion that, while contention I IV's identification of the problem as "the accumulation of mol-i lusks and other aquatic organisms in reactor cooling systems" does not specifically identify any particular type or detrimental i f = t

' + ~ -2 effect of fouling, it broadly identifies the process of fouling. 4. The term "fouling" refers to the formation of inorganic and/or organic deposits on surfaces, and includes that form of L'ouling caused by the attachment of macro-organisms and particulate matter on pipe-liquid interfaces, and corrosion-foul-ing, resulting from the interaction of biological organisms and biofilms or slime layers on surfaces. "Microbiologically induced. 7 corrosion," also referred to as "microbiologically mediated cor-rosion," is one detrimental effect of fouling, which can be caused by the interaction between aerobic and anaerobic bacteria and biofilms in pipe-liquid interfaces. 5. It is my opinion that, because the identification of the issue in NECNP Contention IV broadly identifies the process that causes "fouling" and "corrosion," microbiologically induced corrosion is within the scope of this Contention. 6. "Biofouling" refers to fouling that results from the development of a biofilm consisting of of microorganims and their products (microbial fouling), the deposition and growth of macro-organisms such as barnacles (macrobial fouling), and the accumulation of assorted detritus. Biofouling can be very extensive, even with very minute levels of biofilm. Similarly, i flow or heat inefficiencies can occur even with minute coverage of fouling organisms on surfaces. 7. There are several detrimental effects of biofouling in heat-exchange systems, one effect is blockage of cooling sys-tems, and subsequent impairment of the system's heat transfer

. capabilities. Blockage can be caused by the accumulation of macro-organisms (mussels, barnacles). Another detrimental effect of fouling is contriction and/or mechanical deterioration of the operating characteristics of valves and pumps, which is caused by the accumulation of a biofilm or "slime" on pipe-liquid inter-faces.. Another detrimental effect of fouling is fluid fric-tional resistance, whereby fluid as pumped ineffeciently through pipes. Finally, corrosion and degradation of pipes and heat-exchange systems, as a result of the accumulation of micro-organisms (microbial fouling) on surfaces. 8. The identification of microbiologically induced corro-sion can be seen in Contention IV's use of two words - "aquatic organisms," and "accumulation." The term "aquatic organisms" refers both to macro-organism such as mussels, clars, and other bivalves and bivalve larvae, and micro-organisms, including aerobic and anaerobic bacteria. "Accumulation" is used in the i bio-chemistry field to refer to the accumulation of biofilms on heat-exchange systems, which are the result of aerobic or l anaerobic bacteria depositions, as well as the accumulation of macroorganisms. See Bryers, J.D., Characklis, W.G.,

Zelver, N.,

and Nimmons, M.G., "Microbial Film Development and Associated i Energy Losses," at 12.14-1, Paper No. 12-15 presented at the Proc. 6th OTEC Conference, "Ocean Thermal Energy for the '80's," Washington, D.C., June 19-20, 1979. This detrimental biofilm or l slime can ultimately interacts with bacteria to cause corrosion of heat-exchangers. Thus, "the accumulation of aquatic

~.. 9 -4 organisms" refers equally to the accumulation of microorganisms and the formation of biofilms, as'well as the accumulation of macroorganisms. 9. The problem of corrosion in engineered systems caused by the-interaction between microorganisms and biofilms on pipe-liquid surfaces is not a new one. As early as 1977, 'the cor-rosive effects associated with microbial fouling and biofilm formation have been the subject of studies by the scientific com-munity, and have been identified as a detrimental effect of "biofouling." See Norman, G., Characklis, W.G., and Bryers, J.D., "Control of Microbial Fouling in Circular Tubes with Chlorine," 18 Development in Industrial Microbiolocy, pp. 581-590 (1977), attached as Exhibit E, and studies' cited above. 10. Fouling by macro-organisms, such as barnacles and mus-sels, should not be thought of as independent of microbial foul-ing. Microbial fouling often precedes colonization of heat-exchanger surfaces by macro-organisms, since the microbiological organisms which cause the corrosion are a food cource for bivalves, permitting and encouraging their settlement and colonization, and the sedimentation caused by and causing micro-biologically induced corrosion enables mussels and oysters to attach more firmly to piping surfaces. Therefore, control of i microbial fouling results in control of macrobial fouling. Con-versely, controlling macro-fouling will not necessaryily control microbial fouling or microbiologically induced corrosion. See Characklis, W.G., "Bioengineering Report -- Fouling Biofilm l l i

y l

5-Development: A Process Analysis,".Biotechnoloov and Bioencineer-t ing, Vol. XIII; pp. 1923-1960 (John Wily _& Sons, Inc. 1980), - attached as Exhibit D. M r. James D. Efryers Subscribed and sworn before me this A93ay of February,.1988. m lw e k%, W Notary Publid Mr. Commission Expires April 30,1992 I i i 4 e

I i

i k b L a s h a P - - - ~

Ex.H ^ October, 1967 CURRICULUM VITAE-JAMES D.,BRYERS Associate Professor Center for Biochemical Engineering Duke "niverr.ity Du ...m. North Carolina -27706 USA (919)'684-2434 Telex: DUKETELCOM DURM 802829 Dorn: 19 January, 1951 Houston, Tenas Harried: Christine A. Morgan, 2 sons (Morgan T. and Andrew J.) Home Address: 2915 Friendship Road, Durham, NC 27705 Home Telephone: (919)-489-9885 EDUCATION Ph.D. Chemical Engineering, Rice University, 1990. Dynamics of Early Biofilm Formation in a Turbulent Flow System. Ph.D. Dissertation. M.Sc. Chemical Engineering, University of Idaho, 1976. The Effects of Chain Transfer on Molecular Weight Distributions in an Emulsion Polymerization. M.Sc. Thesis B.Sc. Chemical Engineering, University of Ilouston, Graduation with Honors, 1974 PROFESSIONAL EXPERIENCE A. Teaching and Research Associate Professor, School of Engineering. Duke University, Durham, NC (April, 1985 - Now). Research Scientist (Mitarbeiter). Abteilung Technische Biologie. Institut fur Biotechnologie und die Eidgen6ssische Anstalt fur Wasserversorgung, Abwasserreinigung und Gevssserschutz (EAWAG), EidgenBssische Technische Hochschulen (ETH), Zurich Switzerland (May,1980 - May 1985). Research Fellow. Departments of Chemical Engineering and Biology, University of Calgary, Calgary. Alb6rta, Canada (August,1979 - August, 1980). )

G P8 2 B. Consulting-Oregon Graduate Center, Beaverton, OR - 1986 BIORESPONSE, Inc., Haywood, CA - 1986 Cetus Corporation, Emeryville, CA - 1986 Mestle' Corporation, Yevey, Switzerland 1985 Shell Explora, tion & Production, Aberdeen, Scotland 1985 Ciba-Geigy, Bern, Switzerland - 1983-1985 Shell 011 Company, Westhollow Research Center, Houston, Tx - 1981 Institute of Paper Chemistry, Appleton, WI 1980 British Petroleum, London, England 1980-1981 Shell Oil Company, Calgary, Alta, CAN 1980 Mobil Oil Company, Edmonton, Alta, CAN - 1980 C. Directed Thesis Research Microbiological events in aerobic / anaerobic fouling'biofilms, Ph.D. research of W.F. McCoy, Department of Biology, University of Calgary, Calgary, Alta, CAN - 1982. Transport of groundwater pollutants during peak flood events in the Glatt River, Kanton Zurich, M.Sc. Thesis, EAWJG, 1982. Particulate Effects on the anaerobic production of methane, Ph.D. research of J. Berger, EAWAG 1983. Use of' dynamic tracer methods to evaluate performance of biomass support particles in wastewater treatment systems, M.Sc. research project, EAWAG, 1983. l -Mass transport within biofilms, Ph.D. research of H.

Siegrist, EAWAG, 1985.

l Death, Lysis, and Cryptic Growth in Microbial Cultures, Ph.D. research of C.A. Mason, EAWAG/ETH, 1986. JOURKAL EDITORSHIP Regional Editor, BIOFOULING, Gordon & Breach Science Publ., Beginning 1987 until 1990. PROFESSIONAL SOCIETIES l American Institute of Chemical Engineers Society of Industrial Microbiologists i American Chemical Society - Microbial and Biochemical Technology AWARDS, HONORS, SCHOLARSHIPS Charles P. Schaufus Fellovehip from the Parenteral Drug Association and the Millipore Corporation for research on cell recycle in fermentation systems, Nov. 1985. Award value: $30'000.

.I PUBLICAT. IONS A. Articles in Refereed Journals .Bryers, J. D., Biologically Active Surfaces: Processes Governing the Formation and Persistence of Biofilms, Biotechnol. Proa., 1 (2): 57-68, 1987.

Bryers, J.

D. and Mason,.C. A. Biopolymer Particulate Turnover in Biological Waste Treatment Systems: a Review Bloorocess Enaineerina, 2: 95-109, 1987.

Mason, C.

A.,

Hamer, G.,

and Bryers, J. D., The Death and Lysis of Microorganisms in Environmental Processes, FEMS Microb.olocical Reviews, 19: 373-401, 1986.

Mason, C.

A.,

Bryers, J.

D., and

Hamer, G.,

Activity, Death and Lysis during Microbial Growth in a Chemostat:. Chemical Encineerina Communications, 41: 163-176, 1986,

Bryers, J.

D., Stability Analysis of a Binary Culture Chemostat Experiencing Biofilm Formation, Biocrocess ED9.ineerina, 1, 3-11, 1986.

Hamer, G.,

Bryers, J. D., and Berger, J. Thermophilic Anaerobic Digestion for Sewage Sludge Digestion, ACTA. BIOTECHNOL., E 213-222, 1985. I

Bryers, J.

D. A Structured Model of the Anaerobic Digestion of Biomass Particulates. Biotechnoloav and Bioencineerina, 27(5): 638-649, 1985.

Bryers, J.

D. Biofilm Formation and Chemostat Dynamics: Pure and Mixed Culture Considerations, Biotechnoloav gDd Bioencineerina, 26(8): 948-958, 1984. Bryers, J. D. and Characklis, W. G. Processes Governing Early Biofilm Formation. Biotechnoloav and Bioenaineerina, li (11) : 2451-2476, 1982. Characklis, W. G.,

Trulear, M.

G., Bryers, J. D., and

Zelver, N.

Dynamics of Biofilm Processes:

Methods, Water E.esearch, 16(7):

J207-1216, 1982. i t

McCoy, W.

F.,

Bryers, J.

D.,

Robbins, J.,

and Costerton, J. W. Observations of Fouling Biofilm Formation. Canadian J Microbioloav, 27(9): 910-917, 1981. 2 Bryers, J. D. and Characklis, W. G. Early Fouling Biofilm Formation in a Turbulent Flow System: Overall Kinetics. Water Research, 15(4): 483-491, 1981.

o B. Other Professional Publications

Bryers, J.

D. A Structured Model of Hansenula polvmorcha Diauxic Growth in Continuous Culture, Proceedinas 1987 ASME Winter Meeting, Bioprocessing Colloquium,

Boston, MA.

December, 1987.

Bryers, J.

D. Effects of Cell Recycle on Cell Viability and Metabolism, Proceedinas_ 1987 ASME Winger

Meeting, Bioprocessing Colloquium, Boston, MA.

December, 1987.

Banks, M.

K. and Dryers, J. D. Biopolymeric Particulate Turnover in Biofilm Systems, Proceedinos A.I.Ch.E. 1987 Annual Meeting - Colloidal Phenomena in Biofilm Systems, New York, NY,. November, 1987.

Mason, C.

A.,

Bryers, J.

D., and Hamer, G. Mikrobielles Wachstum in Chemostaten: Ein Tod, Lyse und kryptisches Wachstum'inkorporierendes Modell,

3. Dechema Proc..

Jahrestagun,g der Biotechnologen, Frankfurt, BRD. 1985. Bryers, J.- D., Hamer, G. and Moo-Young, M. (Eds.). Third International Waste Treatment and Utilization Symposium. Conservation and Recyclina, E (1/2), 1985.

Hamer, G.

and Bryers, J. D., "Aerobic thermophilic sludge treatment: some biotechnological concepts," Proceedinos Third International Wasce Treatment Symposium, Conservation and Recyclina, E, (1/2, 1985). t Bryers, J. D., Berger, J. and Hamer, G. Interpretation of Thermophilic Anaerobic Digestion Experiments Using a Dynamic Structural Model. Proceedinas_, Third International Waste Treatment and Utilization Symposium - IWTUS3. Resources and Reevclina R (1/2), Pergamon, 1985. Bryers, J. D., Characklis, W. G.,

Zelver, N.,

and l Nimmons, M. J. Biofouling Film Develepment and Associated l Energy Losses, Proceedinas, 6th OTEC tonference, l G. L. Dugger (Ed.), Washington, D. C., 1979. l Bryers, J. D. and Characklis, W. G. Tha 'athematical l Simulation of Microbial Film Growth. P: oceedinos 97th Annual AWWA Conference, Anaheim, CA, 1977.

Norrmann, G.,

Characklis, W. G., and Bryers, J. D. l The Control of Microbial Films in Circular Tubes with l Chlorine, Developments in Microbioloav, 18, Chapter 48, 1977. l l l l

9 C. Contributions to Boots Characklis, W. G. and Bryers, J. D. Bicfilms in Wastewater Treatment, Chapter 17, in BIOFILMS, W. G. Characklit and K. C. Marshall. (Eds.), John Wiley, (in press). c

Bryers, J.

D. and Characklis, W. G. Biofilms in Biotechnology, Chapter 19. lH:

BIOFILMS, W.

G. Characklis and K. C. Marshall. (Eds.) John Wiley Publication (in press l.

Bryers, J.

D. Mathematical Models of Bacterial Attachment and Subsequent Biofilm Formation. IH: Mathematical Models in Microbial Physiology, Michael Bazin (ed) CRC Review Series, Boca Raton, FL. (in press). Bryers, J. D. and Hamer, G. Microorganisms in Water Purification.Use of Artificially Captured Chapter 11[: Methods in Enzymoloov Series; Enzyme and Whole Cell Technology. K. Mosbach (Vol. Ed) Academic Press, Inc., New York. (in press).

Bryers, J.

D. Application of Captured Cell Systems to Biological Treatment Processes. Chapter 2 in Bioenvironmental Systems, Vglz It, D. L. Wise (Ed) CRC Review Series, Boca Raton, FL. (1987).

Bryers, J.

D. and " L. Irvine. Structured Modelling of biological Treatment Processes. Chapter 6 in Bioenvironmental Systems. Vol. II, D. L. Wise (Ed) CRC Review Series, Boca Raton, FL. (1987),

Irvine, R.

L. and Bryers, J. D. Stoichiometry and Kinetics of Biological Treatnent Processes. 1H: Comprehensive Biotechnology, Volume IV - Principles of Biotechnology: Engineering Considerations, M. Moo-Young, C. L. Cooney, and A. E. Humphrey, Chapter 41, pp 757-772, (Eds.), Pergamon Press, London (1986).

Bryers, J.

D. Biofilm Formation and Its Consequences. Group Two Report. lH: Microbial Adhesion and Its Consequences. K. C. Marshall (Ed.) Dahlem Konferenzen. Berlin, West Germany. January, 1984. Bryers, J. D. Processes Contributing to Biofilm Formation: A Review. Proceedinas First International Conference on Fixed Film Biological Processes, Y. C. Wu et al (Eds.), Kings Island, OH, pp. 155-183, 1982.

1 c. i Characklis, W. G.,

Bryers, J.

D.,

Trulear, M.

G., and Zelver, N. Biofouling Film Development and Its Ef fects on Energy > Losses:. A Laboratory Study, in Chapter 5, Condenser Biofoulina Control, J. F. Garey (Ed) Ann Arbor Science, Inc., Ann Arbor, MI, pp. 49-76 1980.

Bryers, J.

D. and Characklis, W. G. Measurement of Primary Biofilm Formation, in Chapter 11, Condenser Biofoulina Control J. F. Gar y (Ed) Ann Arbor Science, Inc., Ann Arbor, MI, pp. 169-183, 1980. Bryers, J. D. and Characklis, W. G. Kinetics of Primary Biofilm Formation within a Turbulent Flow System, in Fouling of Heat Transfer Eauipment, E.F.C. Somerscales and J. G. Knudsen (Eds.), Hemisphere Publishing Corporation, Washington, D. C., pp. 313-333, 1901. 6 D. Invited Seminar / Conference Speaker: "Biotechnology in Environmental Engineering - Introduction" and "Fate of Genetically Engineered Mciroorganisms in Natural and Envineered Systems," Invited Lecture, American Environmental Engineering Professors (AEEP) Workshop, Philadalphia, Pennsylvania, October 1987. "Modelling of Biological Wastewater Treatment," an IAWPRC Specialized Seminar, August 28-30, 1985. Copenhagen, Denmark. Session Chairman on Basic Kinetics. "Microbial Adhesion and Its Consequences," Dahlem Conference Scheduled January, 1984, Berlin, invited guest speaker. "First International Conference on Fixed Film Biological Processes," invited Session Chairman on Fundamental Biofilm Processes, Kings Island, Ohio, April, 1982. "First International Conference on Fouling of Heat Transfer Equipment," session Co-chairman on Biofouling, Rensselaer Polytechnic Institute, Troy, New York, 1979. I k

S 7 I l-RESEARCH PROJECTS AND ACQUIRED' FUNDING AMOUNT DATE-INSTITUTION PROJECT TITLE. FUNDING SOURCE (in US $. ') 1978 Rice Equipment. Sigma Xi l'000. University, Grant-Ph.D Research Houston, TX Research Society Supplement 1979 . University Anse"robic Province of 150'000, to-of Calgary, Biofilm Alberta, Dept. 1981

Calgary, Formotion in of Natural

' Alberta,CAN Secondary 011 Resources & Recovery Energy Systems 1981 Swiss Mixed Culture Swiss National 200'000, to . Federal Biofilm Devel-Science Funds, 1983 Institute opment: Carbon Water Quality for Water Oxidation and Division Resources Nitrification & Water (Co-investigator: Pollution W..Guj e r) Control,EAWAG, D'd b e n d o r f, Sw i t z e r l a nd 1983

EAWAG, Thermophilic Swiss National 180'000, to DUbendorf, Anaerobic &

Science Fund, 1985 Switzerland Aerobic DiEes-Refuse & Recycle tion of Sludge Division l .(Co-Investi-gator:G. Hamer) y 1985 'EAWAG, Physiology of Swiss National 150'000. to .Dubendorf, Transient Science Funds, 1987 Switzerland Conditions in Microbiology i Microbial Division l Cultures I l 1985 Duke Research School of 15'000. L University Initiation Engineering l Durham, NC Grant c o n t i n u e d - 1 l l L l

A y x s \\=. c 1985 Duke-Research G r a'd u a t e. S c h o o l 12'000. University . Equipment and the Richard-Grant Leach-Research Endovment 1985 buke Research Acadenic Research 5'000. liniversity Equipment Counci'l~, Duke Grant University 1985 Duke Research N Ill Biomedical 50'000. to University Development .Research Grant 1986 Grant 1985 Duke B i o f -f l m Parenteral. Drug 30'000. to University Formation Association &~ 1986 within Cell Mil 11 pore Corp. Recycle C.P. Schaufus Award s Fermentation Systems ~ 1986 Duke Biochemical North Carolina 406'000'. University Engineering Biotechnology Research Center Laboratory D e v e l o p n.a n t (Co-Investigator: H. Clark) i .1986 Duke' Enhanced Enzyme North Carolina 15'000, i: to University Productivity Biotechnology .1987 via substrate Center . Compositional L Transients 1987 ' Duke 1987 Triangle North Carolina -5'700. University University Biotechnology Conference: Center and the l Biotechnology Hillipore Corp. Applied to the Environment 1987 Duke Research Equip-National Science 29'450. l University ment Grant: Foundation Liquid Scintil-lation Counter c o n t i n u e d. 9 -'d s

4-1987 Duke Fate of Biopoly-North Carolina 16'500. University reric Particles Eictechnology in an Aerobic Center Biofilm System 1987 Duke Effects of Exo-Duke University 5'000. University Polysaccharide Research Counci] Production on Biofilm-Bound Bacterial Meta-bolism 1987-Duke Enhanced Enzyme NSF-Biochemical 210'000. 1990 University Productivity in & Biomass Engrg. Micreerganisms CBT-8711612 Experiencing. Prolonged Sys-tem Transients 1987 Duke Workshop on the North Carolina 15'000. University Commercializa-Biotechnology tion of Cellu-Technology l lar Adhesion & (H. Smith-CoPI) Biofilm Processes Total (as of Sept,1987) $ l'495'650. 1 i . J

r 'MICROBI AL FILM DEVELOPMENT AND ASSOCIATED ENERGY LOSSES-i

j.. D. Bryers* *, W. G. Ouracklis', N. Zelver, and M. G. Nimmonsi

+ Entironntental Science and Engineerisg Dept. R. George R. Brot:n School of Engineering ( )(, ([ Rice University Houston, Texas 7 7001 ' Abstract Microbial foultnr. in power plant condensers in-The Problem creases heat transfer and fluid frictional resis-tance resulting in energy losses. Binfouling con-The most corzon method for controlling the fcul-trol is generat*1y'by chlorine addition creating po-ing biofilm development.rd maintaining ^ condenser tential toxicity. problems in receiving waters. A perfornance is periodic chlorination. However. con-better understanding of biciouling film development c e rr. over residual toxicity from hypochlorous acid and destruction ~(i.e.,-stoichiometry and kinetics) or its reactier. Troducts has resulted in federal is necessary to maintain effluent water quality regulations which limit the allovable concentrations while minimizing biofouling ef f ects, of free available chlor!ne in cooling water dis-charges. At the present time, there is no sound ba-This paper reviews research progress in the fol-sis for assissing the impact of the regulations, loving areas: This investigation stems frem the apparent need 1. Development of various sensitive biofilm de-for a more basic understanding of fouling biofilm A tection methods for monitoring the extent of bio-development and fouling biofilm destruction. fouling. Project objectives included the following: 2. Determination of effects of*certain variables 1. Develop a better understanding of fouling bio-on the kinetics and stoichiometry of biofilm accumu-film development, with particular ecphasis on the

lation, ef fects of fluid flow rate, bulk vater temperature. -

'"I ""I#

3.. Correlatien of biofilm de.velopment to in-centration, creases in both heat transfer and fluid frictional rssistance.

2. Determine th'e effectiveness of fouling bio-film destruction by chemical oxidants, prir.arily 4 Determination of the ef fects of chlorine ap-chlorine. I plications on established biofilms. 3. Develop a prack(tal, reliable, suf ficiently sensitive device for monitoring biofouling and for Introduction ef fectively operating arf controlling biofouling de-struction processes at operating power plants. The term fouling refers to the formation of inor-genic and/or organic. deposits on surf aces. In cool. Laboratory experiments and a liraited number of. ing systems, these deposits forn on condensei tube field tests were conducted with two reactor config-valls increasing fluid frictional resistance, accel. urations: .ersting corrosion and impairing heat transfer. Four 1. a tubular reactor cypes of fouling, alone or in combinations, may occur 2. an annular reactor consisting of a stationary outer cylinder and a rotating inner cylinder. 1. crystalline fouling caused by precipitatien of CACO), CaSO4 or silicates The tubular reactor geometry and its turbulent 2. corrosion fouling resulting f rom formation of flow regime are identical to those. existing in cool-insulating layers of retal oxides on the tubes ing water condensers. The annular reactor was test-ed as a biofouling monitor because it is very sensi-3. fouling due to adherence of particulate mat-tive to fe,aling and is easy to operate and maintain. ter on tube surfaces-The annular reactor has the potential of being used 4.- biological fouling resulting from attachmen. in a sidestream frem the cooling water supply to and growth of microbial organisms continuously monf*or biofouling for control of the l addit ion of oxidant. Biofocling in the experimental l This investigation was restricted to the study of reactors.tas reasured by observing changes in the biological fouling. following parameters: i 1 ( 1. biofilm thickness 2. attaeSed bionass j 3. fluid frictional resistance 4 heat transfer resistanee

• Prof essor, Environmental Science and Engineering Dept., Rice University.

proc,,se, in Foullnr. Riof tim Developent se,t,t raduate Assistant, Environment al Science C tit erobial f ouling is the con.bined re sult of mhys-and En:,ineering Dept., Rice University. l l 12.15-1 L db { h C. b 0IW fM Cd, /$pq Ne /2. / { t I m

A o ~x w e, s - = c lilOENGINEERING REPORT Fouling liiofilm Des clopmenI: i A Process Anal sis 3 W (i ClfA R \\( K l. l s. ( ', sn e c, q L y,y,,,,, g I 90 ci o n Bor< man. t /,,,, n, y n yj - ?~ ~- ,p , n_-_=_-_- Summars yQ$[ =" Du film ib t e h mnie m ea sin 1xe is the nti rest,4 id se.eral phs sh il, he n n.il .. ri d mu rohui ri 4sesses m iading t he h uhm m; ilfransr< o t + sidn ids cd.md p.J'n ulate nu:ter s f rom t he Nuk nord to the surf.we. 2) Gr m nos reh.d t eH .a'.n hment 'o t he s urf n, e li Nh r(Ibl.ll f r.in s fi erm.f f h in % f f r(tu Ib re prf %lltClhtrl e's i\\ f' Olq the hh 'II ! M f e s ulllng 'n pr(b Jumon of in g.imc m.o rer di r rhal det is hment or.ne Normi, aue prmu rik io rioid she - ( T hn f erort presem s a trameu or k fo sitco a anah / mg t he m!er't deJ rros e s se s s ont ributmc to b ot'dm Je s clopme n! Some of the asa da He r a t e a nd s ilm r,"n h"i Jara are presented so i that the relans e nriweu rates c.in he ci mr.aed INTR ()lil Cll()N (}" .I.he Ici m louhne iclers to lhe undesirable for mahon of mor can' and or organic derosit s on sullaces l hese derosus can impede the flou s heat across t he suriat e. of ncicase the lhud frichonal resntance al the sur1 ace. and increase the rate of corrosion at the surlaCe in any case' i energy fosses r esuh. Sescral ty pes of fouling and ther, cornbmahons may occur m heat exchangers; il cr> stalhne or precipitation fouh

2) corrosion fouhng.

ne

3) particulate fouhne. O cnemical icaction fouhng. and 5; biological toul-ing or biofouline. Biological fouhne results from al des elopment biofilm con <isting of microoiganisms and tneir pr oducts f microbial foul-of a hi deposihon and grow th of macrooreannins me ).

such as barnacles (macrobral fouhnei. and ci assoiled detruus Ahhough many diffe r e n t mac roorganoim s such as bar nacles and mussels has e been idenhlied m j fouhng communnies. Ihis report w di concentrate on microbial fouhne on the contios ersial premise that n alu a) s precedes colonvahon of the face by macroorgannms. Consequently. control of micromal fouhng re-sur-suhs in control of macrobial fouline. Dt ' elermem of a ss stemahc unJer son hng of Hofouhne fr ern held obsel s a h t nrls has been hmn Cd heCausC ob 'hC m'Cf; sir'In ob % d dn' Bior es h noop s a na n,~e y,,e e r m g \\W x x;jj pg _%,;3;. C 19RIJohn wdes i Sms [nc qg,,, ; A

n I s! I j .[ 1If I ,,i\\ I ,li-t g S n t o n uiy - s r s e t e e tr lmled e imcf t y s i n ma s f b i t a a s n i t te h l. e o y tr b s e u m t u gcih ioin .n 1 t, r u m la u a pn i p t ef n n u J b l e mt c n a l N. n .ule s. a r i l eo a e ish m ason ) m. d e s l m n 1 u l a l h t t u tat l ne tr - u e c c c n gr t cs nnul t h s o I l a n e n r e r c ad s a O. a r n ont nn c cd amet o r s c h n y p o d. a da f w r i lu p p i w n iii e mg i e a. h t n s i h n ahin o c i a eh n s V1 ,o t. c e a ui. e oy a r r t c mni E wEs a. d l iI. h e r Iwll g c c a e g n L n c a D e s m. lu i i l c a u s l co a e ue n a n io

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a

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u. - -.-

u u.J .= p., = = = ,J .~ .,..J r -u ~. o u. .J.,- e 2 r ~ < r 7~ i*. u ?. - w - - .=- . / c" 2 7 = Pt =.. V 3 ~, 3 $,.=e>......x y> -... x... J. .= E. 5 .c.- ~ . = E ; - - x= .~..~ r. -.J w .J k,,. = m .a 2 = o ~ u =.c f y ~u 2 n u

• =

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• J

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v u a u >. n s p

r

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=- r.

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n = =. =. - c .= y l u w= v - c .-i.u = ;- = u : ' ~: -: u - 7 ~g s - n u./. L =- t*

. =

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• J u
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u c i m.x n u.: u-u .., u. o. us= -.a 3 G 5 w;

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c. =..

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• w n C.

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4.

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y, = - - Nn c., y * *' = v.7 7 25 a "cc *

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a

• s C u

- J u.r .u..J d ! J y n \\ x +. - -= ' 7.;, ,J y - .J =- v 2 C. - w / ,J \\ = .r. 2

• =

C.,, Y w ,J e / r .o - -,,,,,,.u e t J - u e U - u ~-. -: o c. J. ~* :e J e .G $ ~.,' u. 23.,=,, 3 "=..3 =. u. - = - u.pu u 7.'2 = = = C ** = y r - C. 3 e :.d.=, 9 3 - -. ,g' y- 0 U d

.=

Vu I w = Z ,g.- -e.";.:= m n y.: -r .=:.=, _ u g 7 C. :,.=-.. r -- J

• J n

C. e C- =.2 3 = 3 "J = w"'8 - f y'-e: -2 .= = Q. G =f,7j.3 ". C - I

- -

4". C

• J z

t w-cx= >= u.J c .U -n ;: c,g -: Y n. w ,=J ". 9 2 ::' = 9 '. =.* 7 u .= c. u = E== 5 = '. s3 '..

• s"' **2 -=

C.,*

w. " =.

. u .,..~.u e k J w# / e -7 . u.=.. C. u^ $ 9.h .- e X : - e ru-A. G ~.5..J. e :: c =. . = w - = = u C." - o .u % =. ;= 5. n 3' P.5 "? c

=

C n - r -- 3 E.2 E E.J 8. = c - J %.~ 9 u.J u n, o -s y - h J J e u -.,X . Ey ~~ _- - {.u..J. = ~ .e -.=. .-,J u u.= E u u.,. e>...= - c ~. -. y... A u -. 4_ -w C.. e

w. -*

a. - o C. 2- - =. =,. = ":: =. =. =,g w .J .r s,, C u - u g.3 4.u t.7

• J
• J v

.J -7 =. = ; 8 * = u,.*: e' u '.: ,g, y g -- y ; w A w W m g = =

• f w

-Jr 2. A. .C = .E a ;c..c v=Ju.3 4 s - 2 O _ # w = v *:

u C. -.,,, -=c

u g "C : 2v cg

= - -= .. c O u u C. v .C b cy e

, - oJ p

a.. s,. u, u

• 3 u r -

-E=J.-

• .= - 6. c

- - u r :* n = .c.3 - 4.C 4 .f w"

J u d-u a w w >i. u'A.-

=. e=s A u J . E u,s w. . w v uC 7 r. C. u c e C C - =.. s u o.u C u uJ 2 ..c C = A e .C . C .s'-.g.A $ c,, e o u o C. ;,-- cw w - e v w 9 ,~ c

• r u ^
• J 2 ir.

.J, u -, u = = .J ,g.- . u t;c u= n u c.n : = === nE p = r e _ d

== c.: s . u ! - =.2-u - c3

O c C. =

... =. - . =. =#'.= Cm.bn.1 J

• J e

s c -

x. - s - u - -

• J w

-a g U - u - c : = .J c w= .u n. u g -s =9 A C..* e / C y p = = ~.. Es ,, u.3 x w )a C r w w = n U +.C *:;. nu - o

• -=..r.

, u =.. ... -Y.: - ': C ,C

• J

.. u ~3 r .J e y -.C onJ ~ C - .J. 2. c c,, - r ... 3 e-y u r .e. g ..,p ,7 A J -, '3 J. J - w - J w C '3 e . A =;% C -( C. '2 -* :* ;,i ~. = - *:

• J = * - u u t., n

-w u

• u 4 --: C - u ^

7 /

• J C. g.. =

w r. T n - =- u.:.:

• J. d. 4 C, g C' *. g

/ as . n. 7 . e u, p = y 3. f .,, r e ,J .A o x -. o q - -* - 4 3. k f .C

*

4 y 'u= a u. c.J e :;;.: w 9 - u u '- %7-8.,,.a n* c.c:. -.; f.J 4.O cf c = G = a: =7 t e o r i;. =I O. '. ; =. 'r 4 c.^ 4 - 2 x g {,= c n : 3 e Gdr = - o e L g J s uou C. i n ~ u 4, ~7.u, O = n. $

• :.: '.:c - 3 :

- = b'" c;,.d ~C .-E,: = = w - c s

1. = =

uw u u C x.-: c =4 wo. d. A. 6 .J -u .u.. r.J w ~. +s1 / rf -. a. 6. u.

,s h w r.

9= 6" n w, 3 t= .J ....=.w w . w '- y *:=

C g.u "3

w n . = :, " = u u .j g - 8 - w n s r.: -U w w.. C..-- . C : "o- .J. .J r w

• C.

.4*.. .J - n C. =. 4 9 7.J / .J C. *%) s -..- . - c ei - > u C t - u -. - >.u C n C - r - J C... - y -. ..,J. - uc g - -u C .= 2 / .n .- r .c.... -* ; - C r u, 7 - *- s 6 = 2,. s*J L l C* f.. e.J ~*.u ,J .g - w p -) 2 .-g {J u .J = ' ~. '- 4.a.

• J

,,, :.~ Y

• a

}. C' n p ^ y -.'Aw.&- =u C. 7 -. c .% 9 ^,. ". ". C J u = J 2 r .J e n .s..J c

r.-

- - 7 -n -7 o.: 2: g C. o..- o.u m j. = u. s : .=..- 7" c

u

.u -u .J ,J e.r= =. -u c :. - !.4 4, ~ .. 2 u u.u = . r .o - ,., 2 y = .r n uw -.-.a - u.

c. 7 :. g w

=

.. N o.3 u

..; y - = w :: = ; ~ - - a .J C. u r - ,- :: =. ~ -o.- .J :. r .J n -u =- c -. - w .u. +u.c- "r

• e

.w... ..=. += =.u w ~ = - i -. 9 ; J, ..u -.:

u. -e e

u u = r 1 = m.. =.9 --c-C. u u= =.. _ =.. u- .u u -: ;- = - -.s _..

g -..-

u. =

x u - d.. ,.t o: - - J m u m ..r..= C..: : = = u: z ..o -= u .u .: 7 u --C,,c. r.. < u - .J u= c u J u

u u,, _ =..*. 7 :

.a C. =,: ?

g:. a.u.

u ;* 0- . =. ~. -.=

u 4

=7 w r u,, 4.?,.. c .u, Ey = g d 'u y,,,=< ' i = d *= =u n J n : c : u ~~. w. u ~: yw. - -. u.. ~ y v <

= 32,- ":

J - : r .J E ~ ' 3 *,: ~~e. uI9C- *? o ~ *i d ,,s. u I c=7 = = u 't - 44 57 E :E 3 3 3

. J i

-. m = d 'E - 3 ' < ;" b.. J - = -=u = : n / 34 = u f- = > G =J g2 ' j 2 { ", =.=. d,".". *M, = y * -c i C. = =~ 4 u ,l':

c. u:

r e 42w I = /.J ~ 7.- n =. r _ a,<,~ < u .J ,J 4" =- - u. ,f *., ' " d } O r u =. : r u-J.- ..e ;.u :- u_z E J c, x- -.- .=. ,u -u- ,J n +y .u ,u xu7=.. u eJ J

• u, s

8 =..J J p. 7 J u = =.,. r .e s -J c p = / = g J u

• . - ;i C. m a

.= k, J. 1, i,* - u . -, - C 1-e. .=.y =_*u,., t w /

• J

= c$ -.' * -* u e 3 u ); w r 7 -b {= 5 ',b.

• • j '

O e r .u. u. y 1 = s C. u7 .~ (.

• 4 u

9 )* 3 fuO O ** )'* $'- u -.E=}= - J 1= su c = - - u .u _-. ':= =.,E -=4 - i o '* = n 3'.=u: e - = = u - J j .e t zS 7 > "':' ,.; i.-=** = " - = =- ?: ~~- u : % 4'J== '"3 1 7 /.= -= ~,,

. =.,

a* = =. -:, <? -y 'J -.) ".-1 "3 -* C. :- u - = = 9

*

C. 4

• • .; =.

= 1 _ =u ~; u:.= / = - u = .e. m /,O 1

• =

^- ~ 3$. s l l \\

l ._-..~.u.~. c ~-> y . e. z u. u s.s.u, , t .s.,,. c s = .u =- . a_u .s-,._.-. m =; ,s o .s u a ~- a e =, r _- c ~. :.,.v g u :u. - c ,= z /n .._ Z... =u..=-

e. e:

-. ?.. .a. .= 3 s .= .u.; .su_u ge m. .1 u.-..=.

c.,.-

e = u w.c 5- -w =.. u Zap .= Ds g .z.. z

~

-u.o, = ~ - m - .a= 3 ., =x xu = .-e E.. =. =:. = c.s : o n.= _c.,.. =. :.=.a .e e: = e n =.. o-x = =. =- c c.

2

. 's 2 u = s o r u-r c n u m .c. - e e - s = -

3 f.:.

.u = s..s .= =. -2 :.a > =.= u u u.=u = u.-.a.: - - - -

r..=.

u. v = e =.=. 'a 3 .u E, -H / - =- - - .s u, Je u2a w i '". n C =c. 'E.' =. = = - c2 :.. d. E.?. 5 5 ~ u u ,,r. s = s e., e: =w = n P. = -u .= c = w a, , s a, =, - ue u u u - - -. =. u .=. - .c.

= e

e _c uu w o = u_ c..= c.- c = .=. a u =. 4- =,.. =.s. u .s : =_- a.,

u. u

.u- - e: =. z s n ; e

== :s = n 2 ~ - ..*_.e .g. .. e ,. -=

r..

.v.. yc m... ~ =. .= = -5, E a=. y sc .c = - 2._x s - - e

• J. >.. -

i ,e .. u _ J .= c - - . = w u .a. - x p -m ~ uEr22 =.=:: s e u - - - >. u - ca - u= 5 r v:.2: 4 = y,s v c u .s. ,u =. -:= .3 - s-w u u > u x,- .v. -. - - - a r .c> =:=_ e .s ~ u c r- ,

• u

..,. n . y c =- c = s=3 ..x ur ~- c. v = = 2 u ~ 3. 3,u ra.-..a = - 9 :- m - u- - > u,=c. u _ = =. - v, u _e a =..u. y.=_ s..= a c..g m e >.u c .s a.- =. ~.u =

=

n o==.cu. n. u .=

i. < -

c, .m .z > c .s . r u. = .= u._ ..s m u ,= ,~4> g. s, e.,, a eo,... - w..,. x c. -. = .u. ,. s...a.g..- s c u ,.e u

e. > -. _-.,.=.- - u.-.-

e ,s = = . = - a -= .a . -. s ,3 ?uu v s t = =, c =.:. -5 p2m= a - - c. :: z- - c> s 4 =. ; u u .~n >u u =: d e, u : u u m... o. -.: :.,s.c, e==.v.ne /. -. m.u, e s= . s& &c, - - - = -3 u = s =, A. .s w -

1. 2 s= 3, 2,.~

u. ;:

y, < l c" 'n u 6. >j u-c d2 's E 3 2 5 ~- ] us. u = _"i u e r-3.5 $. _ v = " 13 6 2:: = s = = :_g,". x, .=. gu q m, u c - s, c. 2 - . c =,3 e., 9 gc ..e - y u / . =. . ~ y,. ~ g e v, : g., =: w -- 5 .J. -c,,..-.. 3y ,J g, g- .. u ic.= ,C n.= p w a x lllll -.- - c .. U y *n f -C w

v -.

t' .s -' - i - 9 < <: = .u. .= ^ C g,." z .g .3 ,,x,,,' o.u. u ..e n s z, e s ,P. u u u = w .. 9.. : uu c

• , C p

e = =.'.W 7 -3 sc = =

• J C

.=.= - w a.:.:' - w f -g=e'>c,, = f 3,, - .Z =. -=. -= - e-e- -,. n ;..,-2 o a =..* u c re J -u as ~~es =. u

*

s . a = - . u"- - e y o u .a. = * -. s s G. 3, = 1 u. a. 1 e y _, =u f.= ,u . =u_ v. ec .u. ~ v a... f. exL-x v.u o,+ x

c. u-

. >*G e . - ~- , c. p- , -... ~ - y o = w e w c. C..s. g u a i - ..A.-. a....,,.- oy,s ,o g -..,s. u,u .s g, s,-,. u c,,, - V z =- -j.E - e z O 3.; y , =-

3. p-un.-

u..f . u,' u, n. f.= ?s - .= = -9 w c c .s .a. r =.. C - C u u U # . 3 i ,r -5 3V = z. E E ~ "3 "u= 3

• 7.Y L o *J"

- j % = u O.= y = .J = A w

• J s

=- =.s:... s s s -?.s / .=. u 2. f.u = uu. -= -2 N 3 ~

..w

- 9 4

• 2 y =

= ~.

.

w , =.

s 3 y

-A t. .~ a.. - e

• Isa

/ - ", T.u .I. = g a:: au y.:ll =- w n. 3.?.s < C, 3 f = f* = = ", c 259 e 33 = 4.u u "d. =.d - = 4 -.Do.C "2 gu - 3 y: 6jO s 3 ~.".e"i 4 E =, u & u: u =s -s t '= Z f 7 -->"J O C = ~~ C

'7,a

~ n. e y* 's e d v- =-=c. = x b, i : :: u, u.- = c = s = u= n x. =, - e u. M.

• f d O

. =

.=aw. C 4 >

> - c .s -o -E ~o< x =e u - 2 >u % w,. c / -e u n - n -s C -, A.c -2 -u ua - u r A o w r 3 - e c' g w se C. 6 3 i' N =1 C s PEP * *. 2 m 3 M 2 8 8 s.'s 5 f,4.*.=E2's H.E i.8't.s Ec21. f ) u J"3 g u

• --.*5 g. as s.

= n M Qy =,=.=.: c,. _ C.~.d. ~- C3..$ u.3.. A, w

3. =

.% c w 3 a oc u = .u. u. - = . ~ - g w w. A O u E .. - w4 u u w...- o

• C,. 2 - 6 t*

C, O. = - A* .,, a n u" C - - o . y "e/ - =, " cu - = c w " 7 .- " O C.

• • b y

r = u L"3 - # '- u, = (,,)

/ C g

u e O e : w n ,J n, -3 +e=.=- 3 {- / =~. c - = uo w u 3. e c e o..e. 2 's =.u .u . = - .=..c. w. : : u c.2 u u w w u 3 -

c. -

= u1 .s, u 'u 6 .a c .e.v.= - .= o s u r.. = o. .v- = - - u .e - 2.n ; w - : e c..: u s

a

.c 7 .o n " ~- =,. u y = .u. . 22 u ,s E = E : .=y U._,.xd >, E.o u p~r x -E.'j 3 ': '.E c.'d

0E

'e a

- n u > ~. u-g u -= ~v-2T r r > s- -E..o, u 9 ..O. -s o E.e r u u o. u u .d. = n ~ =. y o e w>- - -- a ,I 0 $== .c =.=. u-5= s u u= .c ~ S, =.> .,w,.u, .c-c~- v v o u u -. .- n .cs.o <: ~- - >. - e e - .~- -r < a y ay . u s.. c. < w.. - - -- .s 7 o u u.-C u =. C " ., = - c= u - c, c w a -

o

=. n < n u u c ~ - =. = = c". u - ..- = w - : = L, c. l = c C '} C. C.gx w = :. u 0 -

r u, =s - u

$. 3 =u i. Z >~ u; g" a 'u - - -=.=w wC ' =

u

= 0 Ee2 - =4 cd u G ;;T ' ** 9 =, 2 =: u' - - o - % = a g"0 u e / u = ; =.. ,u y '= a.C " u 'i -Ee ??3 i=

. A 'E u

v 4u 3= ug --u y' 4, E, u =u \\ -.,. s - =- = cw - .s n-w n - ~ ~.. .s.. s u - u = I w u 3 ...c - w .r. - c-e.=_ u c _=-- .0 . C_ .s ~u. =, n

c. :

w _.9 .u. - ;,. w u ..9 u - + - * = ..x-n 1, s. 3a='dui 5.., g .s..-s u w.= _,....c - c = w a2

c. 3 n_-

u, r x c. d 0.u ~ u 74..u .a g =u m=,=3

= :

.-7.4~7.: 79 .. r, u G =-- = x u.2 u = 2 =..; ,,. s c c = 5 --u u. e u u E?e.= h 4> .c =v 'e - c - e u u u .o E u 2 C '3..= m.o.E =s

- w u.-

~

G>f =: 3" E = c. = ~- 4 u = =.. ; =.1.= .s u.,.=. sa3 - 2 e e km .= . =. =. r =

= -

= s

.- e-

s c. c c

=w=-,- -,- - -s w .= u -. - ::

c. u. =.=

c. s =. w - - e = 9, .s j - u e.. - y .J,. - - 3 u" u 9 . =. = U c,u -n -= - t>- i f> u r - t E = ;> e=: .= u t-a. =,' =2 w s = -.:,, G :s. 2 = = ~ ~ c, o: r .E : e 5., = s . 9 u=.=,- .s u .a. u, - u u, - .= .l d '" E : - - y L, < o u,r - =- y u -x c-a p. - u 9 -=

c. u 2,

c n. <.*- f s u-c -7 s c = u = s t y -n ~..* a-.A.=. c.J - ~- : u _ -G . u e v n - -.w wy

c. =,

- = = r w - ~ u - - =.j.. w 1 , = -

u. -:: r

.f,. - p a c ;4 "= =

• s e

.J .,= ~.- =.. C s - =.u x +.=: .uc' J _u + s - =. 3 - c u = =,,,= - = v - u e

-=u u

= e s w -. > - uu - =, -a - u -n .:. -- e

o

.-= -9 = r ~:..=, ,y - c, - Cc ,) .=- -3 J -..:: -,r =.-s

e.,.u.n

=. c u_ .a .s. = - -u- , =. r.u :: u w u - y. .s .= t u _u.. 7 -l: -g "j u + 4 =, ". 3 e -u -{ -1 = u4 e r s 7... =. '1O1 - O.= 7- .*i * - - u .c..d, 4 C ; w

• J N#

'g u

• ?*

y. g - _ = - - - u - 4== g = c = G. r - 3 o u

e :

y r

=

,r ;- ; = - u 2 e 3 - n.c a z x v .s, 's 3 A y . s, = ~ <.: - =

c. p

-.=. = .u d .,,".a w= u 0 u = s n s - = v.a = r' u - - - = v ~- - < +- -e 7

*., u =.==J

- = '3 g = 0 4 ~3 ~0

• J

, ' A ' -w ~ ~ -.d=. =

- s 3 c s

-1 = .:- c G. 7 3 3 3* U..d w-7 = - =. - o s

• • O CO

-v. = ~~ ~ .= P I m- ^ m

m...

v \\ ( l a. .2, a.a = = =,. = .t S...y ..J J u =.: .J. . =.. v >>= = -, u, n - - J = .-u ..J s . ~ .r u. ? -.p .J X y ... -n 3_ J .J g- = .J . '-.J

== y =, / , ~,, ,J 2 - = ,,../. u= g - a ,,e .2 .'J e-C

• • • J, A.

.J >J .u 'd = - .J,U* 9 .J J p - r O u :

• • Q "

. C. d [* I / - '..d e ~ o w V

• J

,C

• J n

r o Z = =. = C j.:. - = C C. 2 =. ? . - o - : =. 5_.4. 7 -, n.== . = < u : = =. f.s '.: b C J-C"*.: C " 'u: -u .. u. - ma u = .w a e - >. :_ > - *= 2 - f. .d.

• . E..
  • ..O.

a 6 = ~ ~ .,d,,, X $: ~2 --.n g 9

a.,

• c.

M.y

• E

'A = r l a ? u.,J ^ es*1

• u n

,J u a

• " D.C

> ' ' * =

• .= ^

l .~=,

• 'C

")J""., *g ' '* y J., '5 ~3 .= u C u - . $,. 's -c,.. C $'..*J../ ? u.: ** ~,.~. --.t ~ m J, =

• s

= = g 9.. - y ~ ,.. p u.. .J. ,J= ~. * - ~ ..f J p a.'

• s p.

. 3 9. =1 .J J / -- ~ y ~-.- ~- .J ~,,,,,., ....u .n u_ _ u w w

• e

.-:.v -v* L. # J h- = .: : : -.c. .= u w .._J .J. u, : . - J - u~~...~,. .J - ~ u. p -.u ..J -J b .J = .=.-. :,... .J . u. _o, .4 .~. . yu. - .J A C. = y . u n u.- : r ;.: ui ; n:= J . uu J ..~n ~ ~~. u -

• J,

u- .J _ .u . X -E.2 u p>g g. .J = 9 p, ~ :~e. ~ 3 y.a ::.s.- e g.=, ' g. =.1- .==~. g .o n. .=. c c s..:.?.a= :: f a e5

c.,;.3 _d

, n: 3 = u 2 >=: = 2..:u . _ - u.- s. = = u = J . :.,-.- a. = &. u +- =. 3 .J u .= : - a.:,-s : = 5.J._../y .= J .C, -M ..J. =,-

,37

,.. - - - i.. u ,.n-u, .J c. ..J.

y 7'=.

g - -J - t c s-2. e. * .J a = w O". / M r A =. v c_ :.: 1 C

,

.J G,.'a =,=, - = -, - - u J .: ? = = J n .o j r '~ r r =.y [, .= y n, =J . J

• J L

>..m<Iw = -u. = y x o,-. - o c. u c,o y.: u.

- =.-

u, u > a.~: u =7.:. w '.# d SE ** I ."J

==2 e =,,.g a 5.J u ). *.~: ~ u,,, .s.e. e_ g = ,.,d ,J *- y "J [.( (. S.= y 9 - -, ~.;, =., -..i,. "M ':s + "I 6

• • d A',3 [ O. *..'.J.*- C y'-

= n u,2 *: T

• J C.

/ = .w .= .J e.. .J. -lJ '.. C. "..u., ..,. = * .C. %, .-,.. ~.. .,,,l,.J - / :.c J y 'J C

• O

,a=

• J

~. u w p ? ^ '.J e x ..J.=. e .".e-a Jy. %~ ,..: : u g : s x.. "*.T -w r- : -u-u = 3 m ~J d : s. p u ~. n y r u - = = u e, e o... - a 'n vJ e.-. g,

==J ~.J : ~~~ e.u.i -., y y y J .J y .C ~ U 4*y -.J. : o ,J ~ J e r / .J. g } J

• y = m***

' y,.,,,, = = . C %.j o, ,c,,. u,,, 4.J a

• g

.,.p g e ,3 .J,,. = - -* : p

.- y e.-...

s..* y d G.~~.~/ ~ ~. ~ ~.

• e '.. J ~-

a/ - =...4 ': e.J y y

== s f e .. p .J do C. e { * / 'd U n f 3,y % *e 2.== y ~, i...,, * - nd .J.c "=Ou"

• - 5 *3 =..

=.C .,,,-a'. Q* u ,4

• J

= n w u, ;d' *, ' y =C..s..- - - n - :. = . 'J =.- y

A -

T /

: =

/ .4 O w n,. n .= x u - '-- y . *.J .c., e .u. - u - - r. t,. g - ".- e -w =- .J , p = J n. 7,- 6. ?* F* h * ~ w - A ..g / /. ..U.,,=% y .j C = .C._=..= .J' - C.J ~= g g u A *. ~ eg .e .J. = / * - / ,J b. p==.,...--. = - J -4 a ,e w ? ,I - - u g.".. 'J y (g:3 5. ~,f Y,g ,.i.= .= C

• J 0 5* J u ~ *"* ?.5 5,

.. d f., -Ay f V - b. h k ~5 ! $2 5 ? - ? 5..Q. w.$ = .5. ?"* k $ $". 5, N '_$.?. '] J r J. =..

• d 7

=,

• d ? u,

~ u J u ' = J ? 2 ~= !a y: ;, u, = C : 'c .= i-C. ~ .-~=- u. u =.>..- i w ': n ~ p / - n.u.- y 3 - y _s u Y > y - / ~ /

== .=J =- .J. -,. ~ .%. u u / - c, d...J u: .u. ur u

• • y J
• ,, ' : g

==

• J.... /.,- - y

, 7 .y w- ': y % ,4,,,. 7 : g... g - ? / 't r, ;.,,,.; .., g.,,,. 7 + .s. 2 j u. '=.; e. .J - y f .s.a.y y ~. .J ~=- - y y g,,o y .r - i _J y p - p cs. % n p .J, p g - 'm u -r,u e.E 2.-.=-- n 3, , u >. - -

,u

= u J 7 .. 3. e - ~. -=, y.g g J c 3.(. 4, y :.,3. "" ;. u. =f-6=u5 -ii =,.. x c,, ~ = r ~'., N = =,, =.'d.2 2

• o

< e g, p.. u_ n

t. u *3

= -

7. - g T"'.

J J c J J . C,~ v. l ~ 9 v c ~3 u .-= c-s: C. v,u u - -.J .=. e u, _-.=. r -.. u .J r., .J r

a. v.u

, :. : y .J.- c e... s u = .= ~-

u. h..J.

I =0 ' 7-. ,J J a 5${

• )

.I

5. *

• 9

~- ..-. u C ,"J , **fJ k. *.A.,", . ".. " * * {.. .J,

• f

{ ~g _s '.}J - *} k =. .Jn ~s

• • =
• =..=

.,,, p e J.*

• ~*

~-

• C. -=

="'",9,., 3 a ~~"* *.:. .. C .= = . -= .J p = .. J w l' e-c. r ,=,=,,. =. *J ^ = =J .,.J <~.=y* 1 ** : m e =~ C / .Je

• w
• J g-
• ",a-

.,,,*./. .ie p :

• J

-t u.,,, -* .:f .J - C u

• J,

= , '.= J.*. u-9 ,Y.u,

u,

3 "; / .J y ~=. x.= u G,

• 9 ** C C
• ds l$ n 3

=Un- .,). {. ~ a.-= .: u ** uCM - s.= - u. .'. y 3..C- ^ " * =

• J

-.- g y* / y-g ~ r = 7-g .n > J C~ s =: n.c. .. 7 g -==: :: >. u

• / ? u -

u J.-=..,..G % - 9 aj e -1..' T . =..,. ~ G ,, = '7 3 y -J = n '* 9 ,,l<. . u., c' 9.9 ~~ ~ +J

• J

=.g 'J .c 5 = f E. .u - J - - u : -,. 4 -.- - - -..: - -s ~.: 3, J .J *, 9.J / : =. .a - ';- "4 h, ** .c .u... [*- -.2f ,.4-J 'J'. .L. u,[g a.-.,* ~ 2 - J = .J 7-V O C. f ~/. - b.

• J

.= n 9 - .. I -Cp u O 1.**.'~-

• 7 -* -

a,

• j -* =".J
• J i.J

'I. u d ~* ,J .J e. 6 ""J, 2 t u --E ej y m. ! -g / j .J - =.': ~~ 7.

• 7 C-

-.d.c. c: ,J. J.- e,, _, W *:.. - [ 4. .O.

1. ~

'T O Jp y y n .r

• J
• J

", { V V - sO 'C" ~7*..., = .a

• • =,

J ~*.;U-s j C 'd - ~ ..J. C. J. . e ~. ~ -. .h e :.. ',J =.=:. J u .J .s

• J

$. C.J.*,J- = , J. *{j ** .J.- .J . u ^* J

• d 3 :
• J y

"u J p )*. .3

r.. $., s

,J. ..,.2 -*) ,,f,

• m.)

u a C,J = y ** *"

u. y 3

J -.i y 2 2 ~ n s. ,m. -.J .J .q Q

• • y * :

f. y y ,.,h. - = .y y..h ,/.-. = ~ = ~w.. - - .J, i ,., y eJ .. i

• J
• "l*

f u _..,,.,j p e w .J.e .J /.- r ~- e , y p e = =. e* .s..# - g.. a ,J y .s. J Z J Z. A.*., ,,J

== g .J.,,. ;;- e 2 ~- ,J .J. ~~,, -.,,,, ,g A= Q- = ** - /=,/ / -- J + ,J. = =...-.. y e p p - .J J .J =< - .g* p... .f. y .e. .J f e. .,; * > C J .J 1 -$ f. ? :. ~. *)

• ]

u J ~;. { J

• • . ~ - J,.,-..

w .*, y - 4- '. ,J. / 1 J '- s =. J ./ O.,.- .J

• • C.

<5 .r. C. J .-.J .J. .J =- -. y. 9 =- - .J .J .J .J = .= . + . 3 e. e - g w p

• • • )

w -J w

,== 0 sw a

.. * = - ,O.

• J -A

=, .J. A. .J ,) ae =* O ". ~A .J

• • "3

.,h

• J.

= -

==y

• * /

.V. **./. - ~~~ .r. 0,, - 4 = " e.,, -J f .= ,. =-* =., .f. y ~,,

• J

.) -e "J f = ~ '. C,* 2 - p y

• • • J. **

2 : y s 9 :: / -J ~ p f..

• J *

= f .o dJ. m .J

*.=

.J -. : p - er -. ; -. ,,J,, e 3. e~ C. p* e r a .J J

• R

.d.. - -. .u. * - - J -% .S

f. C.

.- - =. .J .g ./ J. O. ,J -) r 7 <J e-* J -J ~ 4 y ,,"* % eJ w .J. o,/*.** =*..m-r C .J 4 .f = =** t* .J .J = .y .J =- .=,= 1 .J J.'"** J -3.o -J - = - e.~, - y ~* # - y - = O a.

• )
• .J
• s 5, ~

. ~./.,, *" % - J p = J J t - w.** J.. ,7 08 .s .J J e,- . / = - [ j .J J ,. '= - - y _ x r - [ 6 ? ,J S & -J g 1

• a

.. - k.m ~

• J

~ J s. .. -j -== .,J s 9 J - C = - e - . $..,a-2, y - -.y / .n=.3 -. ') : * ~f ':J .O 4- - =* V.O.* = =* / *- =. ** a., .J . -.,,,.* $' $ L s / .J f .J ,J .J - - 2. O u / p O - a.d

• • ~ s -

2 =J 8" . =,, ** w m J

• J J
• 1 ?

!. -..J A ~* J .J. ~- g y V

• ~~.', ~

.n. ~ 2 J, s - y * : " $ -j Q.: C -. ~.J

=

• J r ~

~- ~ .J n -.g R r 1 =)

==o ja =- ,J .J .m,, 9 9. - .J "l" -,.. - a.. u,.. }. =. .J p ., ~ s e e. J y.= - - u. J - - 9 J t = - y y e + J

p. s,ew,,, -

2 -== .O. dC I - t [ .s====... ~. C. = e.- p. [J

== ,j. =*.,. y ~s .J ~ e ' I -.).- P

• 7

-U "J

• ) '. O ~.

I .J .J j .,b I,

• T.

f

• .= -

'd *" / .J d ** '.r.' 2 .=. *-. ,J - ee - y

• )

, = = * * .J .o .J -m - ~ -.= e y a .J J. A. e ') e ~- --. - d r .J J -... e. -' -*

• • • J p -

.) -.Je -.,, .J, . p =. ~,.* .n J, ,,.- -=..j. r .J J p .m. r .J e - a -J t r ~* -.- y,v -

• j -* -
• /

j f .J -1 t. 1- - / *,. - -,J.j -J -r y .f 9.

• J

- - ~., o er .N l. ., =, .s / =

• • ,~

s L* {,"-- *.* .. _ y ', s u .u i ,J .J y,. [ y :.,. i 1 - y op -~O - f -n u_. ,y - O ** =Un

• ,J - : _ -..:,

u-1 ~J J ; 7,. 4: s . -. A O - y '.*.~., 's s S -*: :: -$ 1 = d i ,1 J 1 e / /. e C. e - - - - - - ~ - - - ' ' ' ' ~

gp St I4; titOI.NGINEl: RING RETORT: FOUt.tNG li!Ofit.At DEVEl.OPAtt.-NT 39u m a i so p 4: I 3 ca c... A r a -'S ,f i e j R e movel R.... a s i N (eg/b 4 f' 5 r AFR-Za 2 d 7 I +. + + 2 o O 20 40 60 80 sOO s2O s..eam in.< n...., ra t. j Fig. 7. 1he inih.cnce.4 in..silm thk Loew an t gluc..se 1. =Img :.ne. n gh.ciew seein..r I T ~ tc by b.s hn. Giocow s. nhng e.oc was v v mem' mm' hu At R.Iv.nl luingt 'or AFR 2 8 (Ret. Sil. iii esin.' x 3 1 i 3 2 i f removal rate remains constant (Fig. 71. The critica!. or "active." thick-2 I [e ness is observed to increase with substrate concentration. This behavior I, -3 i is confismed by other investigators 2"' and is attributed to nutrient thf 2 fusionallimitations within the biofilm. Once the biofilm thickness escceds-j 4 c 7 t% j 3 1. the depth of substrate (or oxygen) penetration into the biotilm Wig Kl. 3 5; I; the removal rate is unalfected by lisrther biofilm accumulation. ?' Observed substrate removed rate cannot be used to distinguish between g growth, maintenance, product fermation, and death. It seems clear from l E 'fj 4 ji F other data ' that product formation sprimarily polysacchaside) is signili-2 I j 3 I cant in the c.;rty stages of biofilm formation. Maintenance seguirements i. 3, s - 5 t 3, } or biomass decay become important as the biofilm gets thicker and sub-i g Eg strate does not entirely penetrate the bioliim. These other process rates -i Sg ~e = E %_i= [- 5 w E. e o 25 c. J . j t s " E ~- ni E S 2 3 -+ ~c j r,, j 9 f ;.' g r. c - s c c e l J s f t. - t 39g$2r: 2 Atsy g 5*S: 3 '$ 'f ?35' i 111'133!!~ fA 5 f -54 I r,ssc i u o, ~g-j x E b~2 23 2 i 4 4 JJ 30'E)c62;$k9 c2t

• in -e-0 30j{s"5 w.

i s ( e gsa sa s E7 -- s-s, 3$kh39kN5b a c 3 i ? E. P. E 7 3 7,, u ?.' k c s 2, '5 7,5 7 E 5 = ~ $.g e" u, ,- 2 E T o u.:

• E., *E. 4 Y,

v, : I 8,.E : e' $_ $ 3 2 7 2 =., 3 D!ffuSt0 LEAL RESISTANES !!! h_ $b$ U .f C, - $ $4 r. ~2 'n_7., e m w e 3 -= c 810F it.ftS i i .i J'"

-y # ~.5 6,

67 a a e a a a e a e a j 2 -c s - Fig. 5. As tinitihn thkknew incicases beyinna the depth uf wbsu te hw uugene pen-k >2 1

.7 4 d " " ' I trataan, wbsts.ite hw ogygen) renuwa aare twcomes smicperklent.4 b s

e..lihn thA Luew. l i l ^

1 1 ,4 ) L C -= e 3 E = u .s } i 3 Z / .c E l d' w r 55, l / s-e* = N. .-8 = = 1 \\\\\\ / ~=. =:

5...

/ - m a e = u 2 . i-l l 8 \\ / 4 1 E E i A A wI /, = + -E e. a; r1 e s 2 x e. s v /,= g- + 4 =- = a i E, r,, i { j / W .? 1 c y O y a / z = } / 9 e 1 ? M

• f *

= l' 8 a 1 =- y9 = + 7 4 = Z 5 k l

== u $ / -9:. w a e a g -= 4

/

-2 .3 X9 l ..== 5...: m

= g:

r a - e. w-L + .{ a c,. s .v e - d 3 a / - -a ;s 2 s, r. a2 . z.- - I - ,-j / l_e _. = A 1 ~. c . z = d i

.- s r

2 s

~ 5 i + 6; J 4a .'. g, 9 v-3gr = - 3 i / i !! / 3 l' ~; ' ~ y- ~~. ~ .E' $ 'i / - 'J 'I, 2 !w / i ~.!

• I J

~ 2 .? 'a E 13 $, 3, 3, s -r /, e iI l u e .s >. 3 s r: c=- i .P = T g' y = E L.-=,.'r 7: / C ~ 5 2 =.,3 ; =q d u. :~ c - s r u cn = a - e a - + C .~ d r s e / u L l E 9j ~3 3 'r a u = 0.qdj:!4s 'f-, i s= y w u - c

1. r -

a a / = 2

: e t

.A 2 2.,c 2, p r k a = s r e .' :, c f = 2 -! _t!s i-b ./ -/ r/ 9 5 g.: t v- -g er 0 / e 9 -.-. e - y c. 0 -s 9, 9 (# = c 2 .=: = e y =- = ' a 4, =, =d. y = ; s yv.=, z o a s -9 l = . = s 's =- n -= =- u s /. x .s ' g ': dg ' 1'. '2 : T e r r s = =t -i-. -i 9 -g z -r s = 2 C,. Z, = v. r '- ~3 r l <w = = g - 2.=Ed 2 3. 7 - - r : 2e52 3 =. *.0 ;W 3 v 0 w9 a -. e. 1 2 - 4 - e

- g 2 g -v= =. l 8C 7 -s = a 3 - 3 3 q := :y 1E y ;3 2 <( s vv u

• 3 4

's a e 9 l' l zg = s4 *== r a aw

+

=

2 3 .'.j -3 -3 e a e e e e a e a e s a e a a e l ~. ,3, h N 5 "P 23 4 X," i $ '. > c:* a*: y-i

• *1 - -a; 2,.a. * ;,,. 1 e

s =

• y 4

-e a v = z u v 4 i M M 3 -v ,,,,. u.,.,- y c T- + y'.. e <. es a=a w y. 4. 3 3 7 N c. u e -2. p J -: 7n S g . u. -. /g C -C - = - J 1 2 =. 9 y a9 c. .**v .z o e s. , y /.2.- e g g - 2 .,==. * ~.e = . 'd 6a x V cM .v e. n e, w o ed ,.,= x, = y4

• 3 e

.J g ~ ~r -u4-7 . C..,. J :J W / y .t = w - * = # y a. 3 5,o, d. 4'# rg

• J 9 v p

w "d. j w .O 9 y c,. e - g ). [ , g.j== 7.: O 7 y 2: - o 3 c ::: c o - -

• s f

e Eo,-e .s' u s a, = 3 M e. u.:: e n.c s, =- = a c w. e ..c :., =-: C 2 g -u ~ r ~2 f C _,'u.'.^ - -- - Y =e 0 t' -C e . o n. y" v .C -: -ll + ' e - e e.T., T $.'.7

_s

.: u* - :

oe o s c, s c,,,... - 2 -s 9 C ?,,', r r .=t f< vi

== 4 7a - - 1 ' % s a m- ' e. . O * *. . A > 2 'A V - -. ~: t 7 g a. i ~- f s =

• J v'

• U

- ~- - ~ * - e e s C., V C s, ~ .A x - ; a .: o - - c; '& c & j O g & \\l l '% v s 4 i \\ u., h P' "; : > - e M I s e ;.

==4.p l' \\ .Q .d -~:* 'J'-.=~L 1 i s g J I s x g.: ? i m- ~ 2 - 3 ~- O - s a l s, . o. s g I,, 5 < - ; y4-.i ."s.". O -e _. -.- 7 -2 o r w

• s e=vuu.an Aa e =

e c i J 1 / 2 -e = o 5 I ,= 8 n s e g ,s ~ - =- r l .s, ~ C w3o e ,o T. 3 o / s I. s s, 1

g

= 4 g ;a.,n s o7e p, y e s ,g r 6 =. 3 s g c: y oe O, , o i.s f -C , ~ = s %s v.N 2, , 8e s a i ..= a O e a. s. .e 2 C *. W w g 1 0 tQ, di yi z 5 - '. 0 -4 a2-. o,o %y ev I .g d

• o

- !'4 = -= 90 . 8I? = _s 1, x s w^ 3. shQ.

• d 3 i

-. hy 2 d,s h !3 - 8 4 } ij 4 w. 2 ~ x o c.h...(s ,i N v.2 C. O ,5, o v s o g-3 -,'. :s 3 yd - s 9 e - v t e ,o .o - o o n o,gL o =,4 9 =e 3.3 0 C O A s o o o o o o ~ <., a y, z o,.9"ON_ C.l, = =c y I. '.E.~. 1

f. 3 1

-n i ,e e= s .a r e. = s a a m s 4 - 2 =, 7, -r.2 9 , V - g e -3; as "E e

a

.~ 7 c = 2. d. "-d 22 W- ~ 2E c: -6 a: c-w. 4 y

E-e u

. *b M, 4, - s .Y g .N., .. g 6

• • s b

m 6 .. E{ $d .? 1 ~ >. w cc. e: = t.A Cd. o g.

==. u, .u.. c . u .=.

• u C

. - u e w = = y ,c_.~ = c ~. - E .-m .e ~ w o e. ,,,Q-g e Y- - ' - o U u u (y.,= .u e- = V (,, C "C" Q .: =c.. C xx w -us ~1 Q 2 C. e T T s Ew c. E

• J

- 5 e .:f ,C us a C -~u o, -g .C-g -u 1.C e u ; e r u e a e2. .O -a e C u~ C u -d C- - y .. o./ v y -u -uw ..y* L N .J s. 5 _ y a .J

• d Y..[ d,,,,

g= w, A -54 N La d 2-O O u.C . : o w '.-* C., q -* ( o C u .e.: .= u-n ~- coeu e 'J E 0 = a.=. ~r e, K. c = vw o > / w .y

• e Q

~n u 2# y 0 n m= 2 C. -- C 9**=' = f y g -95

== un Q.' u u.* Z y, w C y - w O -, [J .n -- C.: M 4 C ,/, 4.. ~. - = - m 'i,,.;., g g, -Q y -.A o ..c u n c ., O p 9 . _9 9 ..C .s A u c q. /

s. s - * -

, _ _s a.g - g- / u u c*.,,, _w w y (: c k ..c C J / e v .s + c w y E n, - # u p g / - w.... c -.. u., +.v-

• J
• J 0O&y cA -.-

w* v,w t: w. C.,. u - A -. -E g C. y ; = u A 3e 4_ C."c m C r - e.

== - -- s 7.: .. y. 4. $ .4 u

J C. O O b

.. ~=

n

• a y

n. :- .-w u e g - (,) y w f s .J un y ? 7 u .c - c 'O y* o. U g, yu w - v 0 .J e n..c .J'," c -.t.,. w .e s# a a ~ = s. w n% C C o --*"{ ,d y . e 3 e u u" O C O W 6 'J 1N 5 - C 6 O=- y, 5

79 C.

O n u uM -J to 4 'C w w 4-wE Df =- " A = c. u. 4 - -. =.. c., : - g_. - f .J

u

-d M 9- .C

• J o 9

-r u, C; C s .,,o a. F v; w.,.. ':- 4 +C E.,, u., 2.- ..g .J /7 -u j- -

.= -;

p r. 5 > v / r-.. - y .y w -.: OC f 4 u - / Cw 9

= f c - 4 r,,,,

m g . M.d 7 *: 3 =v = =*d -.J = C -,- 5. " wi.0f T r-y., .= w e 9 V l = 9. :,: . v" .0

T

/

• J e

O., ": - ; - *:

.u

'v .$ O. 2

c. v. : - 2

=.~= O 1 .A w =- 2 -= 3 'G =d., 4 u. C. = - u, =. = v= ~ t. - u - e -4 = A -r. C

• ! e-g

.O. y, y . =. -... 2 p. = .5 x. ~ y . m *,. - o a g , d.. ,.. C

-,'2.C - y~ ~.u u F, d. =2 7

i

~;

i u.

e =. - C C. -. -n. J. C .c = = .r - E= 6

• 2 w

e. .c 3 w x u.. ~ . r -e =,.=- r -.,.s.,- V -- u - a -, 4. 3,: d 3 9 u-u e e - u-e .u, s r sw J C J .: -:.: =- u.. .- :: ? u. l. JI.d. r. : 4G:.: 'E <

u..

C--** 2 3 a < C.u~j.3.y u,y *y,,, q, J -u =.. -s u.J v w I3J G3a= u, e. u = g .5-C. y - s -u - i G 3F, ' -" :53 - -- v s y o. ;J. p ** .l v - 4

4.-.

1

• 1 Q

'e"'.oc o -= u, w. : : v

.=

• = J n

ij 's C 4 " y 5,a N 1 ^ ) ej =$ 5'-

.' 2, c. 4 u-

.=. : - w -e G E 5* C O l l e v - n - . C n C ~ _?> e" - u' u ,? 2 t -s c.C z y: u.-: -v .'. - = u; s.u c. : J ~ u _t j a u.e = =. = v = <

J 9 u

= ', *. f.C - Ae =* -.. i -f = - .J ..~ w* u. - w . - J -o .a s - u ** J v .:. 1 = y 3,v C - J g. * -J .J 9 . *5 'J9 m g g p *f ..,, / .J C C u. c 'J eJ -. 1 y j .$ '} f*} } -y -a .,,,y r -,J . - r w 3 :2 -.2 5 :* 2..h. r'J. 3 'd . b 3" = G i s y ~

. =3.

r s i

• ~>.5,*

I.l [j.'l..b - L. 2,"j 1 - - -u. .J ~.

• C

\\

y - - :

.J - g ~ c : uL. I i.. ~,... .s J d =.=:: v-1 2 - u, - -

.,.=,

a ; e ;5 =...d = -3 ,J

J t u

- u- =.: y : v < - y.J =.: 1 - ?:

.. =z >n_.C 2 : /

. :J n a w i i -

s. >: s (r 4,

= 2 ~ 3 o.7

. 9 3:v J

-.: g: - .a

v =

y : 2

c. :

-C., i 3.3

- f, **,.

:

to r e .J - : - - e^- r a - u-a.u.:. v : - gC" - ' -.d y .= 'J r2 - [ $ :J &..,. o~., ; -, -'J

• W t

. u y C e " e# ', 4 el y. ._,,0 . - 1 v :.. 'Je = **.- ,J v - : 'J J.. Q 2.> E

• V v

.y - g u : g,

-=3

;.=.

.

9.: r J yi

e

=r . ~ W. w w- - - - .-w--. r.-

I e D f f. e I $EYbbh$ .C, O N .L. =u33 .u .= ~./ u ,j .=n.*- d. = u-Z u

• - o 3

ga,, u -4 3 D 2 d 'M,b U e.: S u . 'd

• s X

w *> p c w.. - - 2.=.s s s - t. O 0 n u > =.h

c. c g* c

.m 3 a 1 3 = =-

= =.-

~, e c: v =

r, e

i = :=. :,= = : 5 w s ";; G. a a. n; y : ::u,.==. / d. 2 u = - c. ".x'u c ^ ~3 u 3 u / E u , u ' ** ~3 tyi'E n 2 G C t' >

• 1: < ~;

,i.: E'~> =' / o_L.e4 - P d x s.7 o = o e ' J.. -- =- -= C

• C *

== .C.. *=L u ,= / -e. z s i G w s u* s - _- - u .u*..- J - n - U r = / e m e = / x n=ve d' .= u e.=> v , a u n ,/ l 3 3 m. 3 . g b.- .c a, - uu ou - =. - Z .w - w.= = -u m 5 l 4 3 . = u= c _o,.'> = - 6 w uan 3 c. ,s a 1 g =.== g u..' e =: = = s = n e R. o '.,= s e E.. s e / l. ow / o 1,. u m,=..~. u oy = ~ . = u- .= s

==,k u w 3, i a d g

=

w = n n: / Ya 5 $ $ 5 f 2 'd' / *-O '*I <?g'7$ b 7 .~ * ? *. 5 'G h. n.c. u: E.$. f.s 5: a o : = g z ;g - y. c. =- s I =: w ..e..c i.>. c o = .=. .c m y

e = 7 s Z t ...u e = - c. - . = =, >C >..E -5 p" .'db*.5 m o .=a w *- '2 c. .: u =- -E. y u ~~ ..j 'r. f c*f. E O cu w u a 2 8 g x e' "v A 2-c= .o -=c ee = n...,-.c > w.e, u s - 3 it 9 7 a E 5 u .: -n-e= s A o

• -u-o w

W) $$3NCH1 n11.09 norstm )h i .E - u -E z E= C b 1E a 0-:. g,'2 =: g =.; c.'z. = 5-u.c =. 5 -u.o..c.= =.7.c U., .c. o 0 ,.ea

=.>.: cu o

w o .c. c t.,.o u n -.u n a . u.= -/ u C w eu = r"~' "~,. u u B ,s Ee a.. e :. w,. u.-:.u -=.c C < g '- c - - c: u .g ogu a - c 7.x 3.. - f.:. e a y,, =, c c. = / 4 @.,, C E. t! - ,.=._u n n ,y u .c ? t,, > u, d a n= a ~ u e> c <.c y = E y-- o 5 .=;3 ,.:. =c -, >r u u .1 = i, 4

s w

~c u. n. a n-1, -. ; f $ $ *y g hE b f.h Y 5C '4 = < s y.= = $ ? t-u

==e r

== c= s o e t-E s $ nu a s x0% = . 4 ,s.U N' 3.. ~u. ~ 2 .? l,. C ' -s 5, .- = .2 c ;.fd c 3:'2 = 1 y *- ~: ~c '?.- ~ 4 .= o e l g< ~* u n 2 Rs i e5 6 y72-c.3 [ i l, s = e. a 2 u .=u c my=5y s vn.

Gc;

='bf l -t,g u =u / Q e, e-m = 2 g 2 .: u m - .: u =.x-n-er

c. u,

~~ s .e ~x c. < .s =- ~ b=. u ,,={.,,*. H i-t.5 g E- + cw1 5 i @E s

c.,;: *

c. n j=

\\ i li =y 5 --5 4 uu 3 n. t f =- e.,3 u u g e 5 s ; ~ "z..

1. n-3.

= =.1. " n -t ~/ x ' c.u ' - ~ b r. - y2 s E r HI 5,a!

• t-. r i

52

3..i s i

s - ~ 's n n u. o.w.- - u =. r, a .= r. - --..,x-- ! - c o @2...,. L,, e =3 3. ..a w = _s s., 7. =. .e. s. 3 c. o s ; s es -2, 2 3., - c :;- e

J, y,. t. s n ~* e ki =. : .: - -.d . - c. v = ,s .;a {t. gg -g ;;,,; \\ , g.' ~ 3 ;= = q s ,,J e -J. 1. jg,- i } 4 e ,ee , y =,3 [s - .s j w 2, =v - f ui

• n
• 7

/ s. U

• l*

.O = . - u \\ > r =.. ;. ~ E ~

• 4 -=

..r .c "" *S - [s \\ a . =,, = *.

• J

-O.=.

• s d

C. = t g T -s = s 1 6 =# = O' [. ": C O e e s O ', 7 u -j,Y ~ n e O t g =, -

-

7 { ,s = "" E -=s= 2 f : u u S: .=, - y -= C. C a u. p w-v.- - 7. u [s u -. .t .ue -4 a u = c 5 -7 2'u -- -. = u = =a 7 x4j g. 3 .?. 's'- S- - u u. .> =.-2 ~. So =

f.

• s s
• n e

4 a - s s - s

i 3948 Cll AR ACK LtS filOENGINEERING REPORT: FOUI.ING lilOl'!I.ht DEVEl.OPMEN T tvag .a. 9. / y.. k -q -e 2 0... =, a 5 8 Y e La- $ ss-E e r-6 G 5 .a .- *"I => ^ 3 Initd f.

• 6 8 N/m a

a. A

f.

• 79 N/m' E

e 7

• 150 cm/sec initd i = 180 cm/sec

/ "3 e. n. Emperrnental Run Time (trl Experrnental Run Time (hr) l eg IN Change in prenure drop with umc due to beofilm formahon. Espenment i hg. 20. Ch.mge in a.6 fnciam f ciar ;,nd e a ba.61ni sh. anew wish i,me ti.r an espenmen ...n.1.uca.a sonss no almd selocus s ket 28 C0"'3dC'I di (""ss. ant ps cwuec drop (Rel. 2t <he.aes that Ilim capacity was reduced to 829 of the original capacit)wliere f = l'riction factor (dimer.sionless), d = sube diameter (length). a non h Lib.natos) espeisment.- p = linid density inuas.JlengthS. P = average finid velocity tiengthetimel hNtional iesassance can be represented by a dimensionless fr_ecide = prewure drop along length L (maw / length time ), and I. = length 2 I'.sclos ptsell b) NtWeCII pt'ewill'e pog lg (jegygib t. ,f 4, The change in faiction l' actor and biolihn thicknew w.ith aj,n, imo% I " -.O , !'tytile 2(llor ;t l. abor;gjog y ggghggl3g g g;ggggg.. g)<gLtige !.is Ob%elTCd s.!!!4LLt tll beluviin in a inbid.ir reactor in the liehl gl'ig. 2lt . G. f f..-'~' w N M.- N O o -9 .A.. 3 \\* 3e J To Ec if sr g. 5.; g

f.,

9 2.. v me ..... s. ..........c d Instaat G = 185 crn/sec e..,....., a, I Espersnental Run Time (hr) I l .s e. I y. i., a h..nce in wlunicu ( !)ow s.ste weih anne due to beo6tm forna.ieion. Enpen Tame (days) 6 w.n sona..as J.. unse ne peewus e daop e Rel. 21. ! l'ig. 21. Cli.inge en pscwiere Jmp due to hadilns fium.atum at.a licld Ioeatkus t Ref. 32) h = i

1 1 ,,, - u f- $ 4.y g c. 3 gey ny ;gyy e. 9 5 S.E E. u,w . 5 u c4 o v- = u y u$ d: 05II =2 -e 3 =

a. - =

c - w .3 07 -3 "2 j- >. 7 O d.E ' J :

• 3.

>.. ? a 'n3d:i. 2 =y = ~ = ~ a -u (5 -, e.= u ~s.

4 u.d,

- a e = = ; >V 8. V l-s u 7. c... u -0 c - c wN> - 3o 5 =..t u .=. z ,c.,= >.=. 7. w - =. y.:.d u a.-= x.a u - y.- - =.d = s c

c. x. - >?s =, :

6..

.e. .=. = .< =. c .,b. u.:. o

u.= es

a =.: 9. u -r. =- nw F- .. a.c c i s.c. * >. P o..: c. u -g u. e - s- . - u .,..e,. =.. ~.. L.. M. =u =.- u - *d 3 = /.= =

• k

=

a.-coo o s - ?v .= .s u

c. >.

e -u - = - c e =. > - - o r -5 F. 4== E *. f

=

c. u M

.3 v,,v 5 -Cu= o= c = u. o 2 ~E e "= .M.u92 e

- 5.

.'s u -G w = r v = .x u..k..a, w a c. ,,. u c -< u o e u,,., e v ,e - ~ .3 ~ v - *=. d.:= a ..s'. ~ = < e., / =., 2 u'i e u Y =,. ._.,,u u,, e .J s - C .. 4 ,.."5 .. =. u a ., =..,. .J..a.. J. g- .s. c. =, - c- =-t r -

u

.a : - = .2 y, - - p - g >. u >.: = .-.J t s u, u -.a -e.-

=

== 0 c. ..~. a u x.,. _: . u. u. o u.,,. .n. .a . u cu-u .c .u a - / = =, - c. ,p =. ~ - 2 ?a e n u e .u,,-.". 1 n - - - g s=, u uu =.g / .J %, J,=- .o = =- ww - ei -u.6. u -.,.. - o u _u /- ..=. = T. A .J s o u.4 s ~. - u,. -".=. :. .~ a .. u u ._=. o=.=s --u _= u.. = ., a 2 ....,. : = u e = 2 = c. u = -. -a u .= = c = n,~ m e. - s M - u= =' = 2 - e.,. = R. M =, ',,=.=. >. e. u = v .- d

3. =.

a= ..E. =. g g _ p =w.: q >. 4 63?x e s u e: cP-J u o ud.E.=s.=,2,

d..

y =,3 .= = w3:

c. s x r y.:

u s t _9 '3.cs. 5 c ',.:s,E' M., - = d ._O E.t.5 > u u eg= a 6 u ' E.7 9..:.- y-

c. 3 d

j . s C 2.- .s.a 'e .= s .ucx n,. . 2.u = - a..j a - s 2..a,n,=. u s. s=- ,u =. =- .c,.c o >- o u -. =

u. =

...c _.. s c e., >u 3.5., c c w c c=

c. E.s.s u.:

e F v.E.:,. w s s & sJ =. e a. 9 .c m .J>.>..s u> du. - c J 4

u u

- x --1 uc r e . - u.,. u. r-x Ju-y -

== >.

• 4

....J C. A ;* .r. 7 ,e o .u n - - t mu x..- . =- : u 2.a .u = =~ =- 2e:r.e - p.:= =:.,d.c w n a.=: : a = ..- e- = .: - r u.- =.: u.e UV 3 ~.o 3 o C_ C u. '.c-J 'A.u 4 -$:. .3 9 -e u c G = g ~5 2 c G G 4o.r =o 3.= 2' C sy t- - .= x 2c== - r -c.- - = < =u e - 3=g e =7' o 9c u 1-o: >=- o u= - : j.: n*=lu-E. g.c.

u y2 g, ?

u' u

. :: y =

• d w.e.s g.,

n = .=, ~ A. u 2 u t,. .u -.c = *>~ c.= n= =. - ,e = _= - - = x- - n=s .n c '..s -, - = - =- , c. z.w M.= 9 u eS.c. r = .u i = = u - c i. c,. u s .7.,. .c c c. 2 .e n - - .u=.=- . c, n . c,.; - =. u. u .4 = = = n e =2,,. - >.= y-.= . u z o 5 5 b.c,2. . = ~. = y 9(.==3 .i G u >: : = '5 Y u - 3.=s c' v d= c.c.4 c: = 3.. r,,2. a ~.s- =- o "3 E ~/i 4 >. c. c .s c 2o E5 E=w u.;,= p g = s.,, u .'E *5 G .5 3.y ' =. = .c n u-u - =.. e e s w .- 2 -- / -c = =. E u c w = n <; "O, u5 G,=E. E c '.G 2 -3 4 5 d.e-: - i ~- e - -- u s -s 55 5'd '_n-d.: E ~. u '. ~ 5 ,'E - d C 3 % 'E

4 ~5-y _$

7

d....e c ig 0.:

= = = y. = ': = c rz u s = ~. := 3. - ~ ra ce cea s :d u.4 u u c d:; .=sj.I 4.c "u.

• 2 5

xknac

...; E u.: yC .=um s.c -r *: e o u C -..a.w.-, u.= u r

• n c.

c :v L 1.c u.: u U E4 s-5 - ~ = = u c.: e u y c = 4 y= ~~ u== n.o r L 0

• • a u.d.

=

.= =,,.1

, w "C' e-U c -~ u tu n

• • ',= e n

,*1 h =u E *a s s* V. }u>.r. a nd ** .g g.y N,Lu. Le. n. U u c a .'s.== u".- G .I.

o. c, CT --

o y:,.r u 5 G. u e. c-L o = es c> 3 trg u o=. c u u e . o. go W u.:,..- _ -6 1 5 es*. y d = c g _-. ,e.~ n ? u 'O $E G w a d uj n ,,l 7 le 4 w .=. c w > = / 4-d ~a -u%

• % C l

q g ,,=: e U 2 C*=.: C.:. e e 4, 4 n >- -= w ' = c e $ 2' =x 3 p d Ee. I V 6 -v

- 3 a

o -5 5u - = x = c - - g50u 0 = ? 5 *7 g 05 c w C.- 1,.. 3 5 e j t w e c>=...- p Mx C l y '"

• J -. a>

1 'l c. < 3 m ~; .f .N. '. g =, x p- ,=.: .? 0: ,e a e.#. 4.4 I J, = *. c, ,c, x. u ~ ~- =. + . I2 g, 2 2 5 p 4 u.h u a e v a a ..s .t g-e 7 u v = = u e a ;, c .,. e - 2,, e-x e s,

c:

u f u **,=.g s, s ) 9 7. C - =u-. me .e

• 1, c.

g3 g

• u'

u s o-o :

e, 2 i.w . a u ,s., = 9.c - - = u

• 3 e

7,

8. '1 2: w y

= 2,

~- c =+,

om 9 : u .. =. l .t 7 w~ 9 % e u e w = 3 e

3 2,=.

,4 g -s f /. a .=.

• e>

9 3 3 .s. ', 9 o 7 p C u.., / j v7 = l,p 8 M)I1 '$19Ut/W M DUO $ LYFC" % ~; g d .I v

,::.c,

{ l f1 ; 9 r =i .2.,n= _ wa = =- , u -1 o -f = _/ k) '4 'IIIVd% NO3 N@ f 2,; .= - - -~.M -x 1 2 e = g .=

• • J

=

• l 5

(. 9 - ?a o. 4 - --~$ J f 'e ?. t. < v = = u v

~ I x e a. >I . g i i ly edd lpl a e .n e me n - s e t it ri c s - m a r i e ei i v d m n - s hi n r e s v a l a oi nia te t,i mtoe b r nt d c ef ct g a r r c t a u s n i c E ek ce n ut o s 7 n o isb n C s qe a d "n -s e ef c a v si v ad ai e 2 i e si y c n p b at t et N a nh n nw .m 1 n df a n a l mi A h at o of e r r s c e s s e i a r ei v r s c nup a h u e c o n oh C w i n h i p i r S ct lm (c 2 e g b eif i os T et s n i e s s t s e nl t is toCa d il l a e k a a o n v 0 .a n. I n aif c c S t hi n ic mh g Sn3 t t c somi i inr pm a e 7 n g 3C 0 t a u f E s b a h : t n m.r t e ts 9 u 3 o e mi s a e s t i oc t s r eo yiu2 u l r c n i is in ts t h o= t e or s t s e di oe o a o lmdlie en a t i e s e lmt ov, d r s e > r p n c ?o.r igi o,. v t t E a s y ioib i a r l gaf ih a g ar foilmf o* A s r l i n m l oi w F s eu nt

8 nSmR i s n<

s nn el i f c s n r G xi wOns r a s o u n. b r a nr ea S oa c n b ont imr tc e oi aO ) sx.g o ei g oh opr m N i h n o o i e o wa A n f b f t si s r A tc af s i n a E ( s a. sl e nv nf i T t r n t m e edi tat ed t icCi i t l F ea C e i L f d lafr p .f g I i 4 n t i i f r oi d s r 5 u ub a O h v r e s u e nt s ai T id e f te nd is H ioo n. C T in o r ad ie mnq y s e t h tc s f t a t U kt et m uL tcd a m s. ta ic f iuiob e r it s mnT mi e l u f l n e lei a. r oG r A n e s e s n w t n o u t E s af s r r c ofrb n t ee epf V e o e s a wa grf a ei) f r t s u h al n I a a n w. c tpt e a h I i tad c 4 b x mI ts o c r T n ta e h t n e n n mmc N e e n n. h e n n c e Eg: .l ri n c u n O sh iF r ue r s n wu med a / cit e ini dn s sf e i oiSi a h e s e i r eb d cyoat e f j i n s n r u s M. cma r n s e c l lm H i r n it nTa d .e n qi a n r v wr d n t i v l d wpat h e n e s r

• h t

t os ei e e et a u l / n e os s r f t i o i ab H ei gl F a u cs s l B s o a r u c (s % f mt . i o n toell r imo l mh mt t c t e a s s e n o s e n a O mf ef l j., y., Int 9 n t .a w i a n l nh s adi oiu%t tua nd e9 a e t a eit n tr r e somkn ic l nh l mi f pt a n n r t e I a m t r a tc a h 6. ,jg, IgIia' us .t f nl o.o n r c t a me d t]1 s r ns h i ut f oei O o oot el n r y e oi a v a b rf a h a oi e o o tnti te S e e mipe mtc f l oel h w h f e T vd eh b e r t ma h e c t e e e in inr t y s c uid e d h b a of a r c e n s r v e n v ov s o. nf c r n C d ai e ei hi e mit n s i. s a c r vf t oi E e b. ud omc yf s". f lmd ai s tct mt icd o o s R , gy fm 5t u o ei e x al o m 'd t n ue r e ec e ec ad v e s s mo vt b d s h u t l f i l wc u a r ad 2 c u nd tr u h i lc g nih te te E i c ta nt ocd r h o n o nit a pnl I g t e e 1 n s poet }h!! Q bt c A .s in u. nFt 's a D

a nUl c e

w d ;g hP a C [c /e/8a C[r

• c a

i e l s p 0'l r L vi l o 3 leFtl b e l e 0 n p u m r. r 0m h Zna ma= e p g. s e 0 3 u1 f i c pe y s 0 s i s oy o2 ont r it l wiu lel bllyi a

0. 3 2

e 0n al h r 1 0rp ae a o vb a eT 4 2m 0 *"O 0

n. R l

o c 1O da u a c eu r s lc yd /.T n 6p h a s h 5p 00 = )a.

• r i d s si e

7r 0 0 .4 r s o v s s ubla e = k 0 v e n ,h b w is t e !mu ua r f e o or u e 4, ' k 0 =3, mn r2 lah los i h uo I O s om t c cd s i up = S da s y sb n. /, h o h t s nl f ii J= c/ s aNe o b v v e a.%. 1 / kT we 4 ,h s s oe sao f 0 kT n s f e eich a oJ n 9. k O s ohh e s t o I n n o7 c e et t b ivh a b x y h i n o u e i it oh ch nu t a ~ ei r T r Rl b ^ t e o. ict t k a ci teh l T e oih t w S h mhic t ut r r5 e o. m a n b t e ,. m

o. bs f

3 s a . u i t .l i %'*w lo ns wf n o toe ly 6l er wi2 e e u K o d f t h h N n. a o d N, oe a ,. s n moe t t emwg C n ih s/ s a A ob t d t e r s s c e aNer d i d n. a R r a ~ l . lo \\ ue rc 9 a w s l n e wot r lp n7 e n p

s. n i

n l d tomi mr y y . e k C e et a n e R H .oe i tr n u os to5 ot i c e o n. h d q p po s s raa t r n e s s nti c c r s a e e s sfi e a ey

n. is mhepl niee oh

. a t o t h l e e oih r s nb i s d w:ml o ioll d t c a mr wi t a m .l e l e a as ul i h gi tor a b w so o I e foc t 0 w

o. n x

s n r 4 .a at .r i a r e g r . y i bf e a om e a. et c pdl . o w t pel e e net a a ohl a a ch i u u a t t s a s ~ n n tet mI u. tp t e nl qh i s a n ~ s .cl e i n

• ,~ *

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b n n t a et ih.t e u ci e -. a". woc: s pn v f r. " 3yw

5. E.

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• 3uuru Qo yny.nu out pneoys s! t H h

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3.,

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• • 'ws

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'u., _r _0$0ER NUMBER Q, [ ~ i. The International Corro:,fon Foruni Sponsored by the National Association of Corrosion Engineers / t, larch 22 26,1982 / Albert Thornas Convention Centor, Houston, Texas MICROBIAL FACILITATION OF CORROSION David C. White 310 Nuclear Research Department of Biological Science Florida State University Tallahassee, Florida 32306 ABSTRACT Newly developed sensitive biochemical methods make possible the quantitative study of microbes that facilitate corrosion. At least 3 mechanisms for facilitation of corrosion can now be exauined. The uneven distribution of microbes and their extracellular polymers can create concentrction cells that dif fer in cathodic activity. The netabolic acitivities of aerobic microbes can create anaerobic niches in highly aerobic environments.and from these niches oqganic acids can be generated by the ] activities of fermentative bacteria. In the presence of sulfate or organic ~ sulf ate est.rrs, the oxidation of organic acids can lead to generation of hydrogen sulfide by the sulf ate redsteing anaerobes. Hydrogen sulfide is capable of cathodic depolarization and of oxidation by aerobic Thiobacteria with the formation of sulfuric acid. The detection and validation of "signature" lipids in these microbes can now allow the use of C enrichment 1 Publica tiori Right Ccpyright by the author (s) where copyright is ripphcable. Repro !u:.ed by the !Jational Aesoci.iton of Corto:fon Enginecre with per-rn!3riori of the bothor(,). flACE ha been given first tights of Tsubbchtion of this rna.*u:,cript. ncquests for perrnission 'o publish this rnanuscript h any fonn,in part or in whole, rnust be madein writi~ 11o tJACE, Publiptions Dept., P. O.11ox 210340,llouston,Texcs 77213. The rnanua.cript has nt.1 yct bean revicnc.! sy iJACC., and accordingly, the rnaterial p ct,;ated nod the vice,s cyptessed are Wely those of the author (s) and are not nccer.s: inly ene sed by the Associnti:in. Print <:0 hi USA

y experiments which can provide correlation between corrosion rates and specific microbial activities so that more rational countermeasures can be developed as has been begun for the microfouling community and. the problems of heat t ran s fe r efficiency. I INTRODUCTION With the increasing necessity to recycle both fresh and saltwater, problems of microfouling and subsequent microbially faci'itated corrosion 1 become more important. Not only do nicrobial films increase resistance to efficient heat transfer, increase the resistance to fluid flow and provide the conditions for facilitation of corrosion, but they may provide the ideal growth conditions for the dissemination of the human pathogen Legionella. Simple antifouling treatments with biocides are increasingly expensive and potentially damaging to the environment so research towards a new strategy t0 interdict l microfouling led to the development and validation of biochemical mett.ods by which the biomass and community structure of the microbial films could be examined. These methods can now be applied to increase the basic underst'anding of microbial corrosion facilitation. Microbes can facilitate or initiate corrosion b*

t. heir activities by at least three mechanisms.

Concentration cells differing in cathodic activity can j be generated by the uneven distribution of microbial and extracellular biomass and community compositon. Microbes can generate corrosive cetabolites uch as the organic acids that are important in the weathering process by which soils are replenished or by the generation of miaeral acids under the proper 2 L

a c e ~w t.. conditions of growth. The sulfate reducing bacteria generate hydrogen sulfide with subseanant cathodic depolarization and metalic sulfide formation. With newly developed methodology for "signature" lipids of various physislogical groups of organisms, the microbial ecology of these organisms can be studied to provide a rationale for countermeasures. , EXPERIMENTAL PROCEDURE A flow diagram of the experimental procedares utilized in the study of microbial fouling and corrosion is illustrated in Figure 1. 1 Patchy distribution A coupon of the exposed surface is recovered and stained with aqueous acridine orange. The acridine orange is then vashed off and the relative distribution of intensity of fluorescence measured with an epifluorescent microfluorimeter can give a quantitative estimate of the patchiness as the I specimen is moved across the microscope stage (1). Other coupons can be fixed with glutaraldehyde, dehydrated, coated and examined by scanning electron I microscopy (2). A typical micrograph of the fouling community developing on 'a titanium expopsed to seawater is shown in Figure 2. Lipid extraction The samples are extracted by the one phase-Bligh and Dyer method and af ter inducing phase separation' the lipids are recovered from the organic phase (2). The residue remaining after extraction is removed from the surface by abrasion and analyzed af ter acid hydrolysis. The aqueous portion of the lipid extr, action is also analyr.ed for the adenine nucleotides as illustrated in Figure 1. 3

l 1 Er, F TITLU: Ef f ect of Diofilm Growth on flydraulic Performance-KEY WOi41'S : Biotoulino; Fr ic tional r es is tanec; !!gdr_aulic ' Energy Lonnes ; pjj erolii a ) F i l'p.jd, Pipelines, Sli;no Lavers, Wall lloughncsc, Water Supply ADSTRACT: An experimen tal investigation of the deleterious ef f ect. of microbial slime layecs on the hydraulic performance of water conduits is presented. The underlying mechanisms tha t ' lead to an increase of frletional lossos in the conduit are explored und their relative impor tance is discussed. It is t, shown that although the slime layer is viscoelastic and filamen tous, its ef fect on frictional resistance can be adcouately represen ted through an increase in rigid equivalent sand roughnocs of the conduit wall. -) pMu2 I & & a a D% dsce, /06 (N Y.s~) ff

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e. t .g e ob Ef fect of Biofilm Growth on flydraulic Performance Dy B.'F. Picologlou,' N. Zelver,' an'd U. G.-Characklis 3 INTRODUCTION Biofouling in water conduits causes pronounced increases in fluid frictional resistance. The resulting energy losses arc of major concern to the water. supply and power industries. Biofouling is a general term referring to undesirable effects due to attachment of microorganisms at liquid-solid interfaces. The microorganisms produce a polysaccharide slime layer (5,6,8) which, when formed on the inside surface of water conduits, increases frictional resistance'in flow systems re-sulting in energy losses or losses in pipeline capacity.- +- Deterioration of pipeline capacity attributed to biofilm development can be substantial. Seifert and Kruger (14) report a 55% reduction of original capacity in a 50 mile (80 km) long water supply pipeline 23.62 inches (60 cm) 1D due to a l) thin slimy layer approximately 0.026 inches (650 pm) thick. Table 1 documents other case histories of biofouling in water supply lines (3). Biofouling is not limited to microbial activity. The term includes the interaction of the microorganisms and the slime layer with both the chemistry of the solid surface and the bulk ' Asst. Prof., Dept. of flechanical Engrg., Rice Univ., Houston, Tex. Research Environmental Engr., Cutock, Inc., Sacramento, Calif. 3 Prof., Dept. of Environmental Sci. & Engrg., Rice Univ., Houston, Tex.

P Frictional Losses Due to BiofilmsData Summary from Case IIistories of Closed Conduits Experiencing ~ TABLE 1. Reduction in Conduit Conduit Design Flow Biofilm Thickness Diameter Length conduit Capacity (micrometers) (centimeters) (kilometers) Surface Reference (1) (2) (3) (4) (5) (6) 121 in 2 years 800 105 13 Cement ( 9) 231 1600 90 13 Concrete ( 9) 161 in 3 weeks 3000 90 41 Steel ( 1) 551 in 3 years 635 60 93 Steel (15) 3.53 in 1 year 36 2.5 Steel ( 7) I;ote : 1 micrometer = 3.94 x 10-5 inches 1 centimeter =.3937 inches 1 kilometer .6215 miles = t 9

gy

• ...pr f luid.

These in teractions can enhance some of the more commonly known fouling phenomena such as precipitation or crystallization .v (scaling) and corrosion. In'these latter cases, the wall -layer attains a much more rigid s tructure an d the pronounced increase in frictional resistance can be successfully explained by the increase in the equivalent sand roughness of the pipe

• wall.

In the case of microbial slime layers, the situation is more. com-plex. The thickness and morphology of the slime layers are (_) functions of the operating conditions. A change in operating conditions, such as an increase in wall shear stress, can cause significan t changes in the morphology and thickness of the bio ' film, thus changing the value of the equivalen t sand roughness. In addition, the viscoelastic nature of the slime layer and its filamentous morphology suggest 01at perhaps additional dissipa-tion mechanisms contribute significantly to the increased fric-tional resis tance, consequently, description of the biofilm effect by a. unique value of equivalent sand roughness may be {) inadequate over the entire range of the operating conditions. I The purpose of this s tudy is to explore some of these possibilities. This paper will only be concerned with microbial l slime layers and, th er ef or e, the ter m biofouling will be used for microbial fouling and the term biofilm for the microbial slime layer. EXPERIMENTAL METIIODS s Only the salient features of the system employed ar e given l l here. For additional in formation, see references (16) and (4). i 1he experimental sys tem was designed so that impor tan t l l

7 e ~l', [. J Refvraird frwn Volume is of D WELOPSIENT IN INDUSTRIAL AllCROlllOL OGY A l'u blir curio n of the h<rrry Ivr in.!usr*ial.stu r 4rolwy u 1977 Cal A PT E R 48 Control of Microbial Fouling in Circular Tubes with Chlorine C.NORRHAN. W.G. OI AR ACK Lis. AND 1. D. URYl:RS Department of Environmr ntalScience and Engineering. Rice Uninrsity llouston. Te.cas 77001 hfierobial fouling is a maior cauw of encrgy loss in usier pipelines, heat exchangers, and power plant condenters. Chemical control is usually by chlorine addition. New resirktions on einvent chlonne residuals require that chlorine t'e added judiciously. The uork described is the basis for a methodology to determine optirnum chlonne dosing rates for fouhng control. Tubular reactor esperiments were conducted foi turbuknt conditions (Reynolds numla:t 13.000-19.000). hiicsobial Olm thickness (T) was monitored by elecincat conductivity (12.5 pm) and cortctaicd v ell with increases in frictional resistance measured by pressure drop (6ph Observed op was tigniGeantly higher than predicted based on reduction of crou wetional ares available for now, and a p i, creases of 200% sere observed for T>100 um. Chlorine addiiion caused partial Gim reinovel wii5 consequent increases in einuent particulates. A mathemarical description o' rnictobia! film pe u th and its control by chlorine is offered. INTRODUCTION Microbial fouling is i rnajor cause of energy losses in water pipelines and heat exchangers. Thin microbial Alms attach to the inside of water conduits causing large increases in both Guid frictional and heat transfer resistance. Characklis (1973a.b) and Norrman (1976) reviewed the literature concerning the effects of fouling on frictional resistance. Chlorine Eenerally is use d for controlling microbial fouling in such systems. Hna ever. both economic considerations and increasing.ly stringent en.ironmental regulations require a systematic understanding of microbial fouling,its effects, and methods of control. This caper desenbes researen directed toward the following objectives:

1. Development of a suiuble apparatus for experimental determination of frictional l

resistance as a function of film thickness.

2. Determination of the dependence of fnetional resistance on Gim thickness and now l

rate.

3. Determination of the effect of varying chbrine application rates on film thickness and frictional resistance.

4. Development of mathematical models describing both Gim Erowth and Olm destruction j

by chlorine. i 1 MATT.Rf ALS AND METHOD 5 S>atem description. A lubular reactor was used for reasons of dynamic similarity to full scale l I systems. Figure I is a schematic diagram of the experimental apparatus. Two loops permitted l l simultaneous experiments at difiarent now rates. Each loop cont:uned a rotameter and j l separate sections for Gim thickness, film density, and pressure drop measurements. The entire ) system, including test sections (Fig. 2), was acrylic tubing (1.27 cm 1.D.) roughened te l l . promote microbial attachment. The tuba!ar reactors were operated on a once through basis ( during chlorine addition. i i 1 t SSI i 1 1 l l I I i f l}}