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HomeMy WebLinkAboutNC0050342_Permit (Issuance)_19900726NPDES DOCUWENT SCANNING COVER SHEET NC0050342 Muddy Creek WWTP NPDES Permit: Document Type: ( , , ms,wv mi_i? ham' :: . . ert Issuanc` Wasteload Allocation Authorization to Construct (AtC) Permit Modification Complete File - Historical Engineering Alternatives (EAA) Correspondence Owner Name Change Instream Assessment (67b) Speculative Limits Environmental Assessment (EA) Document Date: July 26, 1990 This documeat is printed on reuse paper - ignore *my content on the reszerge side v FILED, OFF!CF AF AMAIN I4F_ARINGS Jug 26 3 33 FM '90 State of North Carolina Department of Environment, Health and Natural Resources • 512 North Salisbury Street • Raleigh, North Carolina 27611 James G. Martin, Governor William W. Cobey, Jr., Secretary July 26, 1990 The Honorable Julian Mann III Chief Administrative Law Judge Office of Administrative Hearings Post Office Drawer 11666 Raleigh, North Carolina 27604 RE: Winston Salem/Forsyth County Utilities Commission v. EHNR, File No. 90 EHR 0708 Dear Judge Mann: John C. Hunter General Counsel Office of General Counsel Edwin L. Gavin 11 Robin N. Michael David G. Heeter Elizabeth E. Rouse • James C. Holloway John P. Barkley Robert R. Gelblum J. Peter Pascoe, III Enclosed is a copy of the document constituting agency action which, to the best of my knowledge, caused the filing of the Petition in the above referenced matter. Please note my appearance as counsel for the Respondent. Sincerely, • re.„. Eliza•eth E. Rouse Agency Legal Specialist EER/dab cc: Sherry R. Dawson P.O. Box 27687, Raleigh, North Carolina 27611-7687 Telephone 919-733-7247 An Equal Opportunity Affirmative Action Employer ...JUL-26-1990 11:43 FRIOM TO DEM P.02 ncur1 vcv • N.C. Obpt. NRC#Y JUN131990 Winston-Salem State of North Carolina Regional Office Department of Environment, Health, and Natural Resources Division of Environmental Management 512 North Salisbury Street • Raleigh, North Carolina 27611 James G. Martin, Governor Wiliam W. Cobey, Jr.. Secretary Mr. P. W. Swann P.O. Box 2511 Winston-Salem, N.C. 271.02 Dear Mr. Swann: George T. Everett Ph.D. Director June 8, 1990 Mut( 141 4" Subject: Permit No. NC0050342 Winston-Salem Forsyth County Utility Commission Forsyth County In accordance with your application for discharge: permit received on November 28, 1988, we are forwarding herewith the subject State - NPDES permit. This permit is issued pursuant to the requirements of North Carolina General Statute 143-215.1 and the Memorandum of Agreement between North Carolina and the US Environmental Protection Agency dated December 6, 1983. If any parts, measurement frequencies or sampling requirements contained in this permit are unacceptable to you, you have the right to an adjudicatory hearing upon written request within thirty (30) days following receipt of this letter. This request must be in the form of a written petition, conforming to Chapter 150B of the North Carolina General Statutes, and filed with the Office of Administrative Hearings, Post Office Drawer 11666, Raleigh, North Carolina 27604. Unless such demand is made, this decision shall be final and binding. Please take notice that this permit is not transferable. Part II, E.4. addresses the requirements to be followed in case of change in ownership or control of this discharge. This permit does not affect the legal requirements to obtain other permits which may be required by the Division of Environmental Management or permits required by the Division of Land Resources, Coastal Area Management Act or any other Federal or Local governmental permit that may be. required. If you have any questions concerning this permit, please contact Mr. Dale. Overcash at telephone number 919/733-5083. Sincerely, Original Signed By Donald Satrit for George T. Evn ptt Director cc: Mr. Jim Patrick, EPA • AOEY CSSc' Wir so Pollution Preveneon Pays P.O. Box 27687. Raleigh. North Carolina 27611-7687 Telephone 919-733.7015 An Equal Opportunity Affirmative Action Employer JUL-26-1990 11:44 FROM TO DEM P.03 Permit No. NC0050342 STATE OF NORTH CAROLINA DEPARTMENT OF ENVIRONMENT, HEALTH, AND NATURAL RESOURCES DIVISION OF ENVIRONMENTAL MANAGEMENT lEarstal TO DISCHARGE WASTEWATER UNDER THE tlATIONAL POLLUTANT DISCHARGE,ELIMINATION S In compliance with the provision of North Carolina General Statute 143-215.1, other lawful standards and regulations promulgated and adopted. by the North Carolina Environmental Management Commission, and the Federal Water Pollution Control Act, as amended, Winston-Salem Forsyth County Utility Commission is hereby authorized to discharge wastewater from a facility located at Muddy Creek WWTP off Cooper Road Winston-Salem Forsyth County to receiving waters designated as Yadkin River (001) and Muddy Creek (002) in the Yadkin -Pee Dee River Basin in accordance with effluent limitations, monitoring requirements, and other conditions set forth in Parts I, II, and IT1 hereof. This permit shall become effective July 1, 1990 This permit and the authorization to discharge shall expire at midnight on August 31, 1994 Signed this day June 8, 1990 Original Signed By Donald Writ ¶or George T. Everett, Director Division of Environmental Management By Authority of the Environmental Management Commission Ce • :JUL-26-1990 11:44 FROM • • TO DEM P.04 Permit No. NC0050342 SUPPLEMENT TO PERMIT COVER SHEET Winston-Salem Forsyth County Utility Commission is hereby authorized to: 1. Continue to maintain and operate the existing 15.0 MGD wastewater treatment facility consisting of primary treatment using bar screens, grit chambers, and primary clarification followed by secondary treatment in an activated sludge basin and chlorination located at the Muddy Creek WWTP, off Cooper Road, Winston-Salem, Forsyth County (See Part 111 of this Permit), and 2. Discharge from said treatment works at the location specified on the attached map into the Yadkin River (001) and Muddy Creek (002) which are classified Class WS-III and C waters, respectively in the Yadkin -Pee Dee River Basin. 3 z O f fc- O t� A. ( ). F1"rLUENT LIMITATIONS AND MONITORING REQUIREMENTS FINAL Permit No. NC0050342 During die period beginning on the effective date of the permit and lasting until expiration, the Pennittee is authorized to discharge from outfall(s) serial number 001(Yadkin River Discharge). Such discharges shall be limited and monitored by the pennittee as specified below: Effluent Characteristics Flow CBOD5" Total Suspended Residue" NH3 asN Dissolved Oxygen (minimum) Fecal Coliform (geometric mean) Total Residual Chlorine Temperature Total Nitrogen (NO2 + NO3 + TKN) Total Phosphorus Chronic Toxicity"" Qischarge Limitations Monthly Ayg; 15.0 KM 25.0 mg/I 30.0 mg/I 1000.0 /100 ml Weekly Avg, Daily Max 40.0 mg/ l 45.0 mg/I 2000.0 /100 ml Monitoring Measurgment Frequency Continuous Daily Daily Daily Weekly Daily Daily Daily Monthly Monthly Quarterly *Sample locations: E - Effluent, I - Influent, U - Upstream at Idols Dam, D - Downstream at US Highway 64 Upstream and downstream samples shall be grab samples. Requl rements Recording Composite Composite Composite Grab Grab Grab Grab Composite Composite Composite 'Sam, le Locatlori I or E E. E. I E U,D E E E E E E **The monthly average effluent CBOD5 and Total Suspended Residue concentrations shall not exceed 15 % of the respective influent value (85 %) removal. ***See Part III, Condition F; Chronic Toxicity (Cesiodaphnia) P/F at 4.0%, January, April, July, and October. ****See Part III, Condition G. The ; ^ shall not be less that 6.0 standard units nor greater than 9.0 standard units and shall be monitored daily at the effluent by grab sample. be no discharge of floating solids or visible foam in other than trace amounts. e66 Z-9z-1nr' -n 0 0 m m Ql A. (). EFFLUENT LIMITATIONS AND MONITORING REQUIREMENTS FINAL Permit No. NC0050342 During the period beginning on the effective date of the permit and lasting until expiration, the Permitter is authorized to discharge from outfall(s) serial number 001(Yadkinn River Discharge). (Continued) Eftluent Characteristics Discharge Limitations Monitoring Requirements Units (speclfyl Measurement Sample 'Sample Monthly Avg, Weekly Avg, Pally Max Frequency UP& Location Pollutant Analysis"" Annually E - Conductivity Weekly Grab U, D CBC05 Weekly Grab U,D NH3 as N Weekly Grab U,D Fecal Coliform (geometric mean) Weekly Grab U,D Temperature Weekly Grab U,D Lead Quarterly Composite E Lead Quarterly Grab U,D m 0 A. (). EFFLUENT LIMITATIONS AND MONITORING REQUIREMENTS FINAL Permit No. NC0050342 During the period beginning on the effective date of the permit and lasting until expiration, the Permittee is authorized to discharge from outfall(s) serial number 002 (Muddy Creek Discharge). Such discharges shall be limited and monitored by the permittee as specified below: Effluent Characteristics Discharge Limitattopa Monthly Avg. Weekly Avg, Pally Max Flow CB006 25.0 mgll Total Suspended Residue'• 30.0 mgll NH3 as N Dissolved Oxygen (minimum) Fecal Coliform (geometric mean) Total Residual Chlorine Temperature Total Nitrogen (NO2 + NO3 + TKN) Total Phosphorus Cadmium 40.0 mg/I 45.0 mg/I 1000.0 /100 ml 2000.0 /100 ml Monitoring Measurement Frequency Continuous Daily Daily Daily Daily Daily Daily Daily Monthly Monthly Monthly Requirements Samar Type Recording Composite Composite Composite Grab Grab Grab Grab Composite Composite Composite •Samale Location 1 or E E, I E, E E E E E E E E *Sample locations: E - Effluent, I - Influent **The monthly average effluent CBOD5 and Total Suspended Residue concentrations shall not exceed 15% of the respective influent values (85% removal). A discharge may occur only when flow at USGS gage 02116500 located at Highway 64 on the Yadkin River near Yadkin College exceeds 5000 cfs. The pH shall not be less that 6.0 standard units nor greater than 9.0 standard units and shall be monitored daily at the effluent by grab sample. 1 be no discharge of floating solids or visible foam in other than trace amounts. n At-q7-nnr • 0 A. ( ). EFFLUENT LIMITATIONS AND MONITORING REQUIREMENTS ANAL Permit No. NC0050342 During the period beginning on the effective date of the permit and lasting until expiration, the Pemuttee is authorized to discharge from outfall(s) serial number 002 (Muddy Creek Discharge). (Continued) Effluent Characteristics Chromium Copper Nickel Lead Zinc Discharge Limitations Units (specify' Monthly Avg, Weekly Avg, Daily MaX r Monitoring Measurement Frequency Monthly Monthly Monthly Quarterly Monthly Requirements Sample 'Sample Tins Location Composite E Composite E Composite E Composite E Composite E 066 T-9Z-Inr JUL-26-1990 11:47 FROM • TO DEM P.10 PART I . SeetionA ,schedule Q QT ipliance 1. The permittee shall comply with Final Effluent Limitations specified for discharges in accordance with the following schedule: Pernmittee shall comply with Final Effluent Limitations by the effective date of the permit unless specified below. 2. Pcrmittee shall at all times provide the operation and maintenance necessary to operate the existing facilities at optimum efficiency. 3. No later than 14 calendar days following a date identified in the above schedule of compliance, the permittee shall submit either a report of progress or, in the case of specific actions being required by identified dates, a written notice of compliance or noncompliance. In the latter case, the notice shall include the cause of noncompliance, any remedial actions taken, and the probability of meeting the next schedule requirements. II JUL-26-1990 11:59 FRbM TO DEM P.29 G. POLLUTANT ANALYSIS MONITORING CONDITION Part III Permit No. NC0050342 F. The effluent discharge shall at no time exhibit chronic toxicity in any two consecutive toxicity tests, using test procedures outlined in: 1.) The North Carolina Ceriodaphnia chronic effluent bioassay procedure (North Carolina Chronic Bioassay Procedure - Revised *September 1989) or subsequent versions. The effluent concentration at which there may be no observable inhibition of reproduction or significant mortality is 4,0% (defined as treatment two in the North Carolina procedure document). The permit holder shall perform quarterly monitoring using this procedure to establish compliance with the permit condition. The first test will be performed after thirty days from issuance of this permit during the months of January, April, July, and October. Effluent sampling for this testing shall be performed at the NPDES permitted final effluent discharge below all treatment processes. All toxicity testing results required as part of this permit condition will be entered on the Effluent Discharge Monitoring Form (MR-1) for the month in which it was performed, using the parameter code TGP3B. Additionally, DEM Form AT-1 (original) is to be sent to the following address: Attention: Environmental Sciences Branch North Carolina Division of Environmental Management P.O. Box 27687 Raleigh, N.C. 27611 Test data shall be complete and accurate and include all supporting chemical/physical measurements performed in association with the toxicity tests, as well as all dose/response data. Total residual chlorine of the effluent toxicity sample must be measured and reported if chlorine is employed for disinfection of the waste stream. Should any single quarterly monitoring indicate a failure to meet specified limits, then monthly monitoring will begin immediately until such time that a single test is passed. Upon passing, this monthly test requirement will revert to quarterly in the months specified above. Should any test data from this monitoring requirement or tests performed by the North Carolina Division of Environmental Management indicate potential impacts to the receiving stream, this permit may be reopened and modified to include alternate monitoring requirements or limits. NOTE: Failure to achieve test conditions as specified in the cited document, such as minim control organism survival and appropriate environmental controls, shall constitute an invali • tp' '�,� and will require immediate retesting (within, 30 days of initial monitoring event). Failure to: �r �� t� suitable test results will constitute noncompliance with monitoring requirements. ' = o o\ O4k /1. �G O The per iittee shall conduct a test for pollutants annually at the effluent from the treatment p1 The discharge shall be evaluated as follows: g1) A pollutant analysis of the effluent m�, e completed annually using EPA approved methods for the following analytic fractions. (a) purgeables (i.e., volatile organic compounds); (b) acid extractables; (c) base/neutral extractables; (d) organochlorine pesticides and PCB's (e) herbicides; and (f) metals and other inorganics. The Annual Pollutant Analysis Monitoring (APAM) Requirement Reporting Form A and accompanying memo, to be provided to all discharges affected by this monitoring requirement, describes the sampling and analysis requirements and lists chemicals to be included in the pollutant analysis. This monitoring requirement is to be referred to as the "Annual Pollutant Analysis Monitoring Requirement" (APAM). JUL-26-1y50 12:00 FROM TO DEM P.30 • 2) Other significant levels of synthetic organic chemicals must be identified and approximately quantified. For the purpose of implementing this requirement, the largest 10 GC/MS in the purgeable, base/neutral extractable, and acid extractable fractions (or fewer than 10,if less than 10 unidentified peaks occur) for chemicals other than those specified on the APA Requirement Reporting Form A should be identified and approximately uantified as stated in the APAM Reporting Form A instructions. This part (item 2) of the APAM requirement is to be referred to as the "10 significant peaks rule". G 0144,Y QkiL� S 7' b;OMon;ta(.4 ; I wo�(d 2Yo ? P1& wyorry,A July 24, 1990 Mr. Steve Tedder Department of Environment-, Health and Natural Resources Division of Environmental Management 512 North Salisbury Street Raleigh, North Carolina 27611 Subject: Dear Mr. Tedder IQr�02 ne'Mc,Urr li STEUc gate_lG 7 of Winton-6a} rea' " f x d A4 Skr ,14 .,fir yr% 44vc'Y o as g Permit NC0050342 Winston-Salem/Forsyth County Utility Commission Muddy Creek W. W. T. Plant Thank you for the opportunity to meet with DEHNR staff and discuss our concerns about some of the provisions in the above referenced permit. This letter is to confirm our understanding of DEHNR's position on those provisions we discussed during our July. 9, 1990 meeting. 1. Definition of Property Damage: Part II.C.4.a.(2). The City's included a request physical treatment comments on the draft version of the permit that damage to the actual biological and/or processes be included in the scope of the definition of "Significant Property Damage" (Part II.C.4.a.(2)). During our meeting, DEHNR indicated the definition of sig- nificant property damage is per EPA regulations and could not be changed. DEHNR does, however, consider the Dart of the "treatment facilities" and in the scope of the definition. Based existing language is acceptable. "treatment processes" to he therefore they are included on that interpretation, the Box 2511, Winston-Salem. North Carolina 27102 Mr. Steve Tedder July 24. 1990 Page 2 2. Including Water Quality Based Limits Within the Scope of An Upset; Part II.C.5.a. The City previously requested that the final permit include temporary noncompliance with water quality based permit limita- tions within the scope of the definition of an "upset". DEHNR was unable to comply with this request because EPA is in litigation concerning this issue and has directed the states not to modify, the. definition of "upset" until the lawsuit is re- solved. ,In view'toffYthis, the City will accept the current language, but we may elect to apply for a permit modification at a eater .date :to incox.,po.a=t,e any changes in the definition arising as a=.re- sult of the lawsuit. 3. Incorporation of the Provisions of the Domestic Sewage Regulations in the Permit. The fina t:Termit includes several promuigated.�under-_EPA's Domestic Sewage ments of these 'regulations are found in h and Parts III.B.2, 3, 5, and 6 of the conditions?expected - to i*be Regulations.. The require-.. Parts IIIA.'2a. , 2, permit. EPA was to have promulgated these regulations before the ef- fective date of the permit and, consequently, the. City did not. .ob- ject to DEHNR incorporating proposed regulations.. into the -permit. Unfortunately, the EPA administrator has not signed these regula- tions into effect and the final permit does require that we ,comply with unpromulgated regulations. • During our meeting the City of Winston-Salem expressed a re- luctance to accept the permit sections dealing with the Domestic Sewage Regulations. Our unwillingness to accept these areas of the permit was primarily due to the fact that EPA could change the technical requirements prior to promulgation. DEHNR indicated that the EPA had informed them that the regulations would be signed shortly and there was little, if any, chance that the technical requirements would be changed. The City agrees to accept these portions of the permit with the understanding that DEHNR will modify the permit to reflect any changes made by EPA to the final form of these regulations. Mr. St /- v1, Tedder 4. Pretreatment Program Modifications. Our comments on the draft permit included a request that DEHNR stipulate that pretreatment program changes are' minor modifications to the permit rather than modifications. This dis- tinction is important to the City since EPA regulations limit the extent to which changes. can be made when a "minor" permit modifi- cation is made. (See 40CFR 122.63g). During .our meeting DEHNR stated that any modification to the pretreatment program constituted a minor -permit modification in the regulatory sense. DEHNR also stated that minor changes in the plant's,pretreatment program, such as adding a new SIU, would not beaccompanied by the imposition of new monitoring conditions or amended4ef1luent limitations unless thechange could be expected to dramatcaiiy affect the nature of the 'plant's discharge. The City agrees that changing the permit's monitoring and limitatious :requirements;.may be appropriate if a new SIU could be expected `�` `� = adical ly ':alter the p lant's`- discharge. However, we do not wish4to.-have the permit modified to incorporate unwarranted limits :andJmonitoring requirements, or to have unrelated areas of the permit:modified when changes are made to the pretreatment pro- gram. Since DEHNR has acknowledged that all changes to the pretreatment program are minor permit modifications, the City is willing to accept the existing language. We are doing so with the understanding that any changes made to the permit that we feel are unwarranted or in excess of that allowed by the applicable regula- tions may be adjudicated through the Office of Administrative Hearings. 5. Prohibition of Discharges from SIUs Whose Permit Has Not Been Approved by DEHNR. The language in Part III.A.4 of the permit prohibits the City from accepting wastewaters from an SIU until the pretreatment pro- gram is modified. In the City's comments on the draft permit. it was noted that all SIUs added to the pretreatment program are not new discharges. For example, should the definition of SIU become more stringent. many industries could become an SIL without changing the charac- teristics of their discharge. Mr. Steve Tedder J1.11 If the permit language was rigidly adhered to, the City would have to cease sewer service to these industrial users until DEHNR approved the necessary modifications to the pretreatment program. The City feels that such an action would not be appropriate. DEHNR responded that in these situations the. affected indus- tries will be "grandfathered" by DEHNR and the City would not be in violation of the permit or the pretreatment program if sewer service was continued. The City agrees with this approach and the permit language is acceptable. 6. Preparation of Pretreatment.Headworks Monitoring Program. Part III.B.2 of the permii: requires. the City to develop a headworks monitoring program for EHNR's app.roval,.. It was our un- derstanding that'a guidance manual -:was being prepared to assist permittees in developing this program and the City requested that the provisions of this part of the permit not take effect until the guidance was available. During our meeting DEHNRndicated its pretTeatment section had recently made some headworks~tmonitoring guidance information available to permittees and there were not any plans to provide additional guidance. The City therefore agrees to provide documentation on the current headworks monitoring program for the Muddy WWTP. We would like to point out that the permit only allows sixty days from the time the program is submitted until we are required to implement the approved plan. Given DEHNR!;.s. staffing situation and heavy work load, the City of Winston-Salem.is concerned -that DEHNR may need the entire 60 days to review the program. Should DEHNR then require additional changes in the program, the City would not be able to implement the modifications by the deadline. As suggested by DEHNR, the City will include a provision in its program provid- ing for a 6 month implementation delay for modifications requested by DEHNR to allow sufficient time for arrangements. Bioassay Monitoring Provisions and Effluent Limitations. The City of Winston-Salem agrees that it is important to pro- tect aquatic life from toxic discharges. We are not opposed to some for::: of bioi onitoring to protect the receiving stream from toxic discharges. MI-. Steve Tedder The City disagrees, however, that two consecutive failures of the mini -chronic bioassay procedure constitutes noncompliance. The single dilution test protocol is not a definitive test and two tests is a very small data base upon which to initiate an enforce- ment action. In addition, the test can exhibit inherent variabil- ity since the test is a biological assay; the test does not demon- strate the linearity of the dose response; and the data previously submitted by the City demonstrates that non -toxic samples can fail the test. We agree that biomonitoring is an important tool for protect- ing streams from potential toxic impacts. However, we disagree with current. procedures for utilizing biomonitoring data in com- pliance determinations. - During the ineeting we suggested that *a two phase approach to toxicity testing be incorporated into the permit and that compli- ance be based on a six month average chronic effect level instead of two consecutive test failures. A point-:= by point summary of the City'proposal is as fol- lows: Phase I 1. The City would monitor quarterly using the N. C. Mini - Chronic Pass/Fail Protocol. 2. If one test indicates a failure, another test will be conducted within the next month. 3. Two consecutive failures -under Phase I testing would deem the discharge to warrant additional scrutiny and Phase II monitoring begins. Phase II 1. Under Phase II, the City would monitor at least quarterly using a definitive test. The test would measure the NOEC-LOEC interval of a 2%, 4%, 8%, 12% and 16% dilution series. The test result would be reported as the chronic effect level, i.e., the geometric mean of the NOEC and LOEC concentrations. 2. The City would be in violation of the permit if the six month average chronic effect level was equa. to or less than the 7Q10 I. W. C.. 3. The City would resume Phase I monitoring if four consecu— Mr. Steve Tedder tive tests performed at no less than monthly intervals failed to detect chronic toxicity at the 7Q10 instream waste concentrations. The City feels that this approach to toxic monitoring offers several advantages to DEHNR and to the City. A.major advantage under this proposal is that the initiation of an enforcement ac- tion for the excedance of biomonitoring limitations occurs after a pattern of test failures of the facility has been established. The establishment of such a pattern, as well as the use of a de- finitive test to measure the -dose response, significantly reduces the chances of the ..City being deemed noncompliant when it is not and the EPA guidelines for utilizing test data as an enforcement tool ,are addressed.. ''1e'woul'd _also_ like to point -out that a test failure is based on a LOEC value matching the 7Q10 instream waste concentration. Assuming that a test detects the real presence of chronic toxic- ity, an adverse stream effect will occur only if the LOEC concen- tration is equaltoor: less than the actual instream waste concen- tration. Given that average stream flows are much higher than 7Q10, a test .failure 'does not equate to an immediate adverse im- pact cn the receiving'stream and the stream will not be subject to an acute toxic risk while a data base on the discharge is devel- oped. The period between the first phase one test failure and the •point at which the Facility becomes noncompliant could also be used by permittees_to solve their problem without DEHNR having to place them under.a consent order. • During our meeting DEHNR refused to consider incorporating the City's toxic monitoring proposal into the permit. DEHNR indi- cated that an AD Hoc Committee was being established to evaluate biomonitoring requirements and that the modification of the permit's chronic toxicity limitations and monitoring requirements at this time would be premature. DEHNR did indicate that the City could use a definitive test protocol, but the current policy of defining noncompliance as two consecutive test failures will remain in effect. The City of Winston-Salem feels that even with the use of a definitive test protocol the two consecutive failure definition of noncompliance is unacceptable in that it may subject the City to an unjustified enforcement action and/o: a ,.:itizens suit. Mr. Steve Tedder t age In that the Ad Hoc Committee is to begin work very shortly in reviewing biomonitoring requirements, we suggested that the toxic- ity limitations provisions of the permit should be held in abey- ance until action on the committee's report is taken. DEHNR stated that they would not agree to hold these provi- sions in abeyance, citing the need to be uniform in their treat- ment of permit holders, and an extension of the thirty day permit appeal period is not possible. Given.these conditions, the City of Winston-Salem has filed a petition with the Office of Administrative Hearings requesting an adjudicatory hearing in this matter. If-DEHNR is agreeable, how- ever, . we will request that an indefinite stay be. granted by- the administrative law judge to allow time for the Ad Hoc Committee to complete their review before the. idjudicato ry petition proceeds or the City decides to dismiss it. Please contact me at your earliest convenience if our under- standing "of DEHNR's position on the. items discussed during our July 9, 1990 meeting is inaccurate. You may reach me at 919/784-4700. I would like to thank you again for your time in assisting us in this matter. Your efforts in explaining DEHNR's rationale in preparing this permit have helped resolve most of our concerns. We sincerely appreciate your cooperation in this matter. Sincerely, 'Stan_ . Webb Plant Manager -Muddy Creek Plant pc: Crystal Couch, IWC Sherry Dawson, Asst. City Attorney It _t) JuN 2 2 1990 TEC'iNICAL PPORT BRANCH State of North Carolina Department of Environment, Health and Natural Resources Division of Environmental Management 512 North Salisbury Street • Raleigh, North Carolina 27611 James G. Martin, Governor George T. Everett, Ph.D. William W. Cobey, Jr., Secretary Director June 19, 1990 Stanley B. Webb City of Winston-Salem Box 2511 Winston-Salem, NC 27102 Subject: Proposal For a Wet Weather By -Pass City of Winston-Salem's Muddy Creek Wastewater Treatment Plant Forsyth County Dear Mr. Webb: In your letter to Mr. Trevor Clements of my staff dated February 28, 1990, you requested that the Division give you an indication as to our position on the possibility of installing a wet weather by-pass at the subject wastewater treatment plant. It is my understanding that the City is concerned that the influent pumps to the treatment facility will be damaged if they continue to have to pump more than their design capacity. In your letter you indicated that this by-pass would be on a very infrequent basis and only at times of very high stream flows. While I appreciate your concern for the protection of the facilities influent pumps, I can not be supportive of the by-pass of untreated wastewater from the Muddy Creek facility or any other facility in the state. I realize that your facility and many others in the state experience challenge when wet weather results in much higher than normal flows into your systems. The Division's position on this matter is that efforts must be made to reduce infiltration and inflow to levels where the problems are eliminated or the permittees must construct additional units at the treatment facility to be able to handle the increases in flow. I realize that this in many cases is an expensive undertaking, but at the same time it is one that the citizens of North Carolina expect and deserve. I hope that this will clarify the Division's position on the issue of by-passes. If there is a need for any additional information or clarification, please do r hesitate to contact Mr. Dennis R. Ramsey of my staff or myself at 919/733-5083. S` erely, eorge cc: Steve Tedder Dennis R. Ramsey Winston-Salem Regional Office Pollution Prevention Pays P.O. Box 27687, Raleigh, North Carolina 27611-7687 Telephone 919-733-7015 An Equal Opportunity Affirmative Action Employer MEMO TO: S�eJC� epe,.fi Gis Sam 50 &AA. tA-4- cA. e �c LO-s DATE• SUBJECT: pcQ 1� )SSuse_ �J ; �-e d' ri Gf Cj North Carolina Department of Environment, Q Health, and Natural Resources otr4- PUBLIC WORKS DEPARTMENT City of Winton-a1em April 30, 1990 Mr. Dale Overcash, NPDES Permits Group North Carolina Department of Environment, Health and Natural Resources Division of Environmental Management Post Office Box 27687 Raleigh,. North Carolina 27611-7687 MAY 0 (99O PERM!7s FF �� C4.it • •n •S RE: Comments on Revisions to Parts I, II, and III of the Draft NPDES Permit for the Muddy Creek Wastewater Treatment Plant; Permit # NC0050342; City of Winston-Salem, Forsyth County Dear Mr. Overcash: Included herein are the comments the City of Winston-Salem wishes to offer regarding the revisions to Parts I, II and III of the draft NPDES permit for the Muddy Creek Wastewater Treatment Plant (N. C. 0050342). These revisions were received by the City on April 2, 1990. Part I.B.2: Requirements for Operation of the Facility at Opti- mum Efficiency. This provision of the permit requires the City to provide the operation and maintenance necessary to operate the plant at opti- mum efficiency. The City of Winston-Salem requests that DEM delete this part of the permit because its intent is addressed in Part II.C.2 and its language is inconsistent with the requirements of 40CFR Part 122.41(e). Part 122.41(e) requires only that facilities, be oper- ated in a manner to achieve compliance with the permit conditions. Box 2511, Winston-Salem, North Carolina 27102 Mr. Dale Overcash April 30, 1990 Page 2 Part II.A.5.a: Definition of "Monthly Average Discharge." It appears that a portion of the second sentence in this part of the permit has been inadvertently deleted. This sentence should be changed to read as follows: "It is, therefore, an arithmetic mean found by adding the weights of the pollutant found each day of the month and then dividing this sum by the number of days the tests were re- portede Part II.C.4 a.(2): Definition of "Severe Property Damage." The permit defines "severe property damage" as being substan- tial physical damage to property, damage to the treatment fa- cilities which causes them to become inoperable, or substantial and permanent loss of natural resources which can reasonably be expected to occur in the absence of a bypass. As worded. the definition of severe property damage does not appear to include damage or disruption to the physical and biological treatment processes occurring in the treatment fa- cilities. In as much as there are occasions when it is necessary to by- pass actual treatment processes to prevent a prolonged period -of noncompliance due to their disruption, we feel that the definition of severe property damage should include damage to treatment pro- cesses, as well as, damage to the physical structures housing these processes. The City of Winston-Salem requests that Part II.C.4.a.(2) be modified to read as follows: "Severe property damage" means substantial physical damage to. property;. damage to the treatment facilities, including disruption' of the treatment processes to the extent that a violation of the permit limitations can be expected to occur, or substantial and permanent loss of natural resources which can reasonably .be ex- pected to occur in the absence of a bypass. Severe property dam- age does not mean economic loss caused by delays in production. Mr. Dale 0vercash April 30, 1990 Page 3 MAY v ;96 Part II.C.4.b.: Bypass not Exceeding Limitations. PERMijs & _ I p:ty� i7ii <n This part of the permit allows any bypass to occur which does i not cause effluent limitations to be exceeded, but only if it is for essential maintenance to assure efficient operation. The term "efficient operation" is not consistent with the provisions of 40CFR 122.41 e, which require only that the permit - tee properly operate and maintain the facility to achieve compli- ance with the conditions of the permit. The City of Winston-Salem requests that the term "permit com- pliance" be substituted for "efficient operation" in the first sentence of Part II.C.4.b. Part II C.5.a.: Definition of "Upset." The permit defines an upset as an exceptional incident wherein there is temporary and unintentional noncompliance with technology based effluent limitations because of reasons beyond the control of the permittee. The City of Winston-Salem feels that the upset definition should also apply t.0 ii; i t.' ..._1i. 1:1.._ ;,Il:lans which cater quality based. It is entirely conceivable that a permittee could unintentionally violate water quality based limitations due to the occurrence of an exceptional incident beyond the control of the permittee, i.e. "upset." The City of Winston-Salem requests that the first sentence in Part II.C.5.a. be changed to read as follows: "Upset" means an exceptional incident in which there is unin- tentional, and temporary noncompliance with permit effluent .limitations because of factors beyond the reasonable control of the permittee. Part II.C.5.b.: Effect of an Upset. The first sentence of this part of the permit references paragraph (3), which does not seem to apply to "upset." Further, there appears to be a word deleted from this sentence. We request that this sentence be changed to read as follows: An upset constitutes an affirmative defense to an action brought for noncompliance with permit effluent limitations if the requirements of Part II.C.S.c of the permit are met. Mr. Dale Overcash April 30, 1990 Page 4 Part II.C.6 : Removed Substances. pER - - ;i jiv This part of the permit requires the City to comply with all existing federal regulations governing the disposal of sewage sludge. The City feels that the purpose of this clause is addressed by language in our sludge disposal permit. No regulatory need is thereby served by including this provision in the Muddy Creek Plant NPDES discharge permit and we request that the second sen- tence in this part be deleted. Further, this part of the permit provides for a reopener clause to be incorporated by reference into our sludge disposal permit. The reopener clause allows DEHNR to reopen our sludge disposal permit to incorporate the applicable requirements of 40CFR Part 503. We object to the inclusion of this language in the permit be- cause it essentially modifies the conditions of our sludge dis- posal permit without providing for our rights to due process, and it allows DEHNR to reopen the permit for a reason which is not specified under 40CFR 122.62. EPA regulations specify the situations under which permits may be reopened to incorporate new standards without the permittee's consent. The need to incorporate revised sludge dis- posal standards is not one of the situations wherein EPA allows a permit to be reopened by the permitting agency. We request that Part II.C.6 be revised to read as follows: Solids, sludges, filter back wash, or other pollutants re-' moved in the course of treatment or control of wastewaters shall be disposed of in accordance with NCGS143-215.1 and in a manner such as to prevent any pollutants from such materials from enter- ing waters of the State or navigable waters of the United States. The permittee shall notify the Permit Issuing Authority of any significant changes in its sludge use or disposal practices. Part II.D.6 : Records Retention. This part of the permit requires the City to maintain all records for a period of three years. It is further provided that the Director may extend the retention period at any time, -upon re- quest. Mr. Dale Overcash April 30, 1990 Page 5 R tfri;EIVED MAY 0 3 i90 Whereas the term "at any time" can be construedto include the time frame after the three year retainage period had expired, we feel that the permit language should be modified to require the Director to make an extension request within the three year pe- riod. read: We request that the last sentence of this part be modified to This period may be extended by request of the Director at any time within the three year record retention period. Part II.E.5.b. Monitoring Reports. This provision provides that if we monitor any pollutant more frequently than required by the permit, the results of this monitoring shall be included in the calculation and reporting of the data submitted in the DMR. 0ur previous permits contained language which restricted this requirement to that monitoring done in excess of the frequency re- quired for the specific locations delineated in Part I.A. of the permit. We would like to see the language in the new permit so writ- ten to prevent our having to routinely report the results of the extensive amount of "in -plant" monitoring the City does forpro- cess control. We request that Part II.E.5.b of the permit be modified to read as follows: b. If the permittee monitors more frequently at the desig- nated locations than required by the permit, using test procedures specified in Part II.D.4 of this permit, the results of this monitoring shall be included in the cal- culations and reporting of the data submitted in the DMR.` Part III.A.2.b.: Requirements for Control of Pollutants Attribut- able to Industrial Users. A key word appears to have been inadvertently deleted from the sentence. Please modify the language to read as*follows: Mr. Dale Overcash April 30, 1990 Page 6 MAY U 3 690 b. Wastes which will cause corrosive structural damage to treatment works, and in no case, discharges with pH less than 5.0 standard units unless the system is specifically designed to accommodate such discharges; Part III.A.4.: Pretreatment Program Submission or Modification to Include SIU's. The provisions of Part III.A.4. require the City to modify our existing pretreatment program per 15NAC 2H .0907(b) prior to accepting wg,stewater from any significant industrial user. The City of Winston-Salem is concerned about the possible ramifications of the use of the term "modification." Section II.B.14 of the permit provides that pretreatment program modifica- tions shall be considered as permit modifications, thus opening every permit provision for possible change each time a new SIU is permitted. The City feels that the addition of new SIUs to a permit'tee's approved pretreatment program is actually a "minor modification" as per 40CFR 122.63g. Federal regulations provides that the per- mitting authority is not allowed to modify existing permit provi- sions or incorporate any new provisions into the permit when mak- ing a "minor" permit modification. We request that the last sentence of Part III.A.4. be modified to read: Prior to accepting wastewater from any significant industrial, user, the permittee shall either develop and submit to the Division a Pretreatment Program for approval per 15NCAC 2H .0907(a) or request a minor modification to an existing Pre- treatment Program per 15 NCAC 2H .0907(b) and 40CFR 122.63(g). The permittee shall not be required to prohibit discharges from existing industrial users, being reclassified, as a significant user, while DEHNR is reviewing the SIU's proposed permit. We also request that the existing language in Part .I1I.B.14 be modified to read as follows: Modifications to the approved pretreatment program, including but not.limited to local limits modifications and monitoring Mr. Dale 0vercash April 30, 1990 Page 7 program changes shall be considered a permit modification or a minor permit modification depending on the applicable pro- visions of 15 NCAC 2H .0114 and 40CFR 122.63g. The City would also like to point out that the permit does not contain a definition of Significant Industrial User. We feel that this definition should be included in the permit to prevent any confusion as to which industrial users need to be included in our Pretreatment Program. Part III. B.: Pretreatment Program Implementation. The City of Winston-Salem is concerned that DEHNR may have made significant changes to the State's pretreatment program with- out formal EPA approval, as required !by 40CFR 123.62. • The City does not feel that any such unapproved changes are federally enforceable under the NPDES permit. We request that a statement be added to Part II.B. of the permit to read as follows: Pretreatment program requirements incorporated into the permit which have not been formally approved by EPA as per 40CFR 123.62 are not deemed to be federal requirements and are not federally enforceable under this permit. Part III.B.2.: Headworks Monitoring Program. The permit requires the City to submit a headworks monitoring program proposal to DEHNR for approval within 120 days of the ef- fective date of the permit. DEHNR has been developing a guidance document to assist per- mittees in developing such a program but it has not been released. We object to being required to formulate a headworks monitoring .program until DEHNR's pretreatment section provides this guidance document. Part II.B.3.: Headworks Analysis. Since the City has not had an opportunity to review the head works monitoring guidance document, the Clty reserves the right for future comment after having adequate opportunity to review the guidance document. Mr. Dale 0vercash April 30, 1990 Page 8 Part III. B.5.: Monitoring and Inspection. MAY u /99b �-r'rl)) 'flif.� r. The last sentence requires the permittee to inspect all SIUs annually. Currently there is not statuatory authority in either state or federal regulations for this requirement. We request this requirement be deleted, or revised to be consistent with NCAC 2H.0916 (c)(3)(c). Part III.C.: Authorization to Construct. The City of Winston-Salem has previously requested that the language in this section of the permit be modified to make it workable and consistent with federal requirements. As .written, the permit requires that the City undertake no construction or additions to the facility without written approval from DEHNR. We feel that this requirement is unworkable in that minor modifications must be done routinely to effectively operate the plant, and neither the City nor DEHNR has the manpower to pre- pare and timely process the plans and specifications for the num- ber of changes that would fall under this broad permit provision. Moreover, the D. C. Court of Appeals has held that EPA lacks the authority to impose a construction ban under the NPDES pro- gram. NRDC vs EPA, 822 f.2d104 (D. C. Cir. 1987). We request that the provisions of Part III C be limited to those construction projects undertaken to add to the plant's treatment capacity or to those projects intended to change the type of process used in the plant's system. We also request that wording be inserted into this part of the permit stating that, in view of the 1987 decision by the D. C. Court of Appeals, this re- quirement is not federally enforceable. Part III.F.: Chronic Bioassay Testing Requirement. The City of Winston-Salem requests that the language in this section of the permit be modified to reflect the pertinent re- quirements governing quarterly biomonitoring for acute toxicity (Reference Part 1 of my March 30, 1990 letter to you). The City does reserve the right to comment on any permit language proposed for this section regarding Acute Toxicity biomonitoring. Mr. Dale Overcash April 30, 1990 Page 9 Part III.G. Monitoring of GC/MS Peaks. Part III.G. of the permit requires that the ten largest GC/MS peaks in each organic analytical fraction be quantified and iden- tified. We have previously requested that this language be modified to limit this requirement to those GC/MS peaks, not to exceed ten, which can be identified with a confidence level of at least ninety percent (90%). DEHNR has previously indicated to the City that it would evaluate the language and change it accordingly. The current ver- sion of the language is still unchanged from that originally pro- posed. We request that the language In the second sentence of III.G.2 be changed to read as follows: For the purpose of implementing this requirement, the largest ten GC/MS peaks in each organic chemical analytic fraction (or fewer than 10, if less than 10 unidentified peaks occur) which can be identified with a confidence of at least.90%, for chemicals other than those specified on the APA Monitor- ing Requirement Reporting Form A should be identified and approximately quantified as stated in the APA Reporting Form A instructions. The City of Winston-Salem sincerely appreciates the opportu- nity to work with DEHNR in finalizing this permit. We are willing to meet with DEHNR staff to discuss our comments or this permit. Please contact Mr. Stan Webb at 919/784-4700 if you wish to' schedule a meeting or if you have any questions. Thank you for your cooperation. pc: It Sin4erely, TILIITIES DIVi• ION Tom Griffin,'' . E. / Utilities Superint: %ent Crystal Couch, IWC Supervisor Stan Webb, Muddy WTP Superintendent Dr. George Everette Larry Coble, NC -Regional Office DIVISION OF ENVIRONMENTAL MANAGEMENT April 6, 1990 MEMORANDUM TO: Dale Overcash (\--ry THROUGH: Trevor Clements FROM: Ruth Swanek Vs, SUBJECT: City of Winston-Salem Muddy Creek WWTP NPDES No. NC0050342 Forsyth County I have reviewed the letter submitted by the City of Winston- Salem for its Muddy Creek WWTP. The following changes should be made in the draft permit: 1. The monitoring frequency for lead should be changed to quarterly. 2. You should read through items 4-19 of their letter to determine if the new boiler plate will address these items, and if it does not, whether anything should be changed. 3. Item 15 of their letter addressed the GC/MS peaks monito- ring. We should send Winston-Salem the new language and reporting sheets. I am preparing a cover letter for Steve Tedder's signature which should be sent out with the revised draft permit. If you believe that anything in items 4-19 should be addressed in the cover letter, please let me know as soon as possible. This cover letter will address certain issues raised in their letter which need additional explanation, and state that this permit represents DEM's final stance. We should prepare a permit package for Winston-Salem which includes the following: 1. Cover letter 2. Revised NPDES permit 3. New boiler plate 4. Ken Eagleson's document which contains DEM's responses to the questions most frequently asked about our toxicity program. (I will attach this to the cover letter which I will send to you). When the permit package is complete, the entire package should be sent to Steve Tedder for his review and signature on the cover letter. If you have any questions, please advise. cc: Steve Tedder Steve Mauney Central files State of North Carolina Department of Environment, Health, and Natural Resources Division of Environmental Management 512 North Salisbury Street • Raleigh, North Carolina 27611 James G. Martin, Governor George T. Everett, Ph.D. Wiliam W. Cobey, Jr, Secretary DirectorApril 25,1990 Mr. Tom Griffin, P.E. Utilities Superintendent Public Works Department City of Winston-Salem Box 2511 Winston-Salem, NC 27102 Subject: Muddy Creek WWTP NPDES No. NC0050342 Forsyth County Dear Mr. Griffin: A modified draft NPDES permit and a public notice that have been prepared for your facility by the Division of Environmental Management (DEM). The draft permit (NC0050342) contains modifications made in light of your March 30, 1990 letter requesting several changes, and the permit as written reflects DEM's final decision regarding these matters. For clarification purposes, the following additional explanation is offered: 1. Toxicity The chronic Ceriodaphnia toxicity test will remain in the permit. Mr. Ken Eagleson of our Environmental Sciences Branch recently provided your office with a list of sources which contain the basis of North Carolina's tox- icity procedures in. his letter of February 1, 1990. In addition, I have enclosed a document containing DEM's responses to the questions most frequently asked about our toxicity program which I believe will satisfy your concerns in this regard. 2. Lead Monitoring DEM made an error in preparing your last draft, and the monitoring frequency for lead has been changed to quarterly. We apoligize for any inconvenience this may have caused you. 3. Outfall 002 The limits and monitoring requirements for outfall 002 will remain in the permit. DEM understands that that your operators have no control over when the plant discharges. However, to ensure that this outfall is only used dur- ing rare storm events, discharge from pipe 002 will only be allowed when the Pollution Pie endon Pays P.O. Box 27687, Raleigh, North Carolina 27611-7687 Telephone 919-733-7015 USGS gage 02116500 located on the Yadkin River at Highway 64 near Yadkin Col- lege indicates that the flow is greater than 5000 cfs. According to USGS records, this flow corresponds to approximately the tenth percentile of flow at this location. That is, the flow should exceed 5000 cfs 10% of the time. The flow at the gage during Hurricane Hugo was greater than 12000 cfs, and flow data indicate that the flow has been greater than 5000 cfs on numerous occassions when you did not discharge through outfall 002. The metals and nutrient monitoring for outfall 002 will remain in the permit. We expect that the loadings will be much higher during storm events and the data will help us evaluate any impacts. DEM is interested in assess- ing short term acute impacts as well as long term impacts caused by a dis- charge. 4. Monitoring of GC/MS Peaks The language in this permit condition has been slightly revised. DEM is working on a new reporting form which will include columns for the compound identification (i.e. library match, retention time, etc.) and the probability of a correct identification. If a compound cannot be specifically identified, a compound class may be listed instead. DEM will forward you a copy of the new form upon completion if you request it. You will find that many of your concerns are addressed through the new boiler plate language (copy enclosed). Any requested modifications not made in the permit reflect the final view of DEM. If you believe that your con- cerns have not been adequately addressed, you have the right to an adjudica- tory hearing upon written request within thirty (30) days following receipt of the final permit. This request must be in the form of a written petition, conforming to Chapter 150B of the North Carolina General Statutes, and filed with the Office of Administrative Hearings, Post Office Drawer 11666, Raleigh, North Carolina 27604. Unless such a request is received by this Division, the requirements contained in this permit shall be final and binding. If you have any questions, please contact Don Safrit or Trevor Clements of my staff at (919)733-5083. Attachments cc: Don Safrit TWIsmoreqftemett's Steve Mauney Central Files Sincerely, Steve W. Tedder, Chief Water Quality Section PUBLIC WORKS DEPARTMENT City of Winton- Salem APR 0 3 )990 March 30, 1990 PERMITS RI != f,INEFRIN(; Mr. Dale Overcash. NPDES Permits Group North Carolina Department of Environment, Health and Natural Resources Division of Environmental Management Post Office Box 27687 Raleigh, North Carolina 27611-7687 RE: Comments on Draft NPDES Permit for Muddy Creek Wastewater Treatment Plant; Permit No.: NC0050342; City of Winston- Salem; Forsyth County Dear Mr. Overcash: The City of Winston-Salem wishes to offer comments regarding the draft NPDES permit for the Muddy Creek Wastewater Treatment Plant (NC 0050342) which was released for public comment on Febru- ary 20, 1990. This permit is being reissued following our request for a permit modification dated September 15, 1989. The City of Winston-Salem and DEHNR have negotiated several changes to the permit's boiler plate which do not appear in the draft. Parts I and II of the permit are not included in the draft document, although DEM stated that they were going to change the language in several of the provisions in those parts. The City re- serves the right to comment on provisions and requirements con- tained in Parts I and II of the permit, since these parts were not included in the draft. For the purpose of commenting on the draft, the City of Winston-Salem is offering "new" comments regarding this document and we are also summarizing those changes DEM has committed to make in the permit which were not included in the draft. In addition, we are restating the basis of our objections to other provisions in the permit that were noted in our letter to Paul Wilms of September 15, 1989 and which have not yet been re- solved. Box 2511, Winston-Salem, North Carolina 27102 Mr. Dale Overcash March 30, 1990 Page 2 Since more detailed discussions were included in the letters dated September 15, 1989 and January 11, 1990 the City hereby in- corporates those concerns and objections contained in the two pre- vious letters in lieu of reiterating each point in this letter. 1. Comments on Effluent Toxicity Permit Limitations The draft permit requires quarterly chronic toxicity testing of the plant's effluent and defines noncompliance as two succes- sive test failures. The permit requires that the bioanalysis testing be performed using the N. C. Mini Chronic Bioassay Proce- dure. The City of Winston-Salem does not feel that it is appropri- ate for DEM to impose effluent toxicity limitations on the plant's discharge. DEHNR and EPA have determined under Part 304 (L) of the Clean Water Act that there is no need for Winston-Salem to ad- -;_ ,kdress or to eliminate water quality based toxic pollutants. DEHNR it]N �fshould, therefore, require only monitoring for whole effluent tox- -I icity. 3 �v Furthermore, the City of Winston-Salem feels that the N. C. Mini -chronic (Pass/Fail) bioassay procedure does not accurately predict reproductive impairment in Ceriodaphina dubia, DEHNR's required indicator organism, and that the test is imprecise. 0n these bases the test procedure should not be used as a tool to de- termine permit compliance. The City of Winston-Salem reached these conclusion following our review of test data collected from mini -chronic bioassay analyses performed by certified labs across the State after June 1, 1988. The detailed discussion of this study is attached as Ap- pendix I, entitled "Chronic Bioassay Concerns and Supporting Data." The results of the study are summarized below: 1. 43.5% of samples which failed the test based on reduc- tion were declared toxic when they were not. Nine (9) such samples were statistically compared to sixty-six (66) other control groups. Four of the nine samples, declared toxic, would have passed the test at least 74% of the time when compared to other control groups. 2. In five (5) of six (6) control group replicates, the test predicted chronic toxicity when there could not have been any difference in the incubation water and, therefore, no toxic components present. Mr. Dale 0vercash March 30, 1990 Page 3 3. In 36% of sixty two samples passing the test, the control would be judged toxic in comparison to the sample dilution being tested. This indicates that a "beneficial factor" is present in the sample and not in the control water. Test failures may, therefore, be caused by the absence of a beneficial factor in the sample rather than the presence of a toxic substance. 4. Reproduction in Ceriodaphnia dubia is impacted by normal water qualities including hardness, pH, con- ductivity, and osmotic pressure. The test's protocol does not include procedures to negate these factor's influences and, therefore, the N. C. mini -chronic bio- assay procedure is not a controlled test. 5. The indicator organism required for use in bioassay monitoring by DEHNR has a cyclical reproductive pat- tern which is influenced by factors that are not yet understood. 6. The N. C. mini -chronic procedure is accurate only when the sample tested fails due to mortality. In this res- pect, the procedure is no more accurate than the Acute Toxicity Test. The City of Winston-Salem does not wish to accept any permit provision which could result in an enforcement action based on the results of a test which we feel is inaccurate. The City requests that the permit include provisions for quarterly effluent monitoring only, using the Acute Toxicity Test. 2. Monitoring Frequency for Lead In a September 15, 1989 letter to Mr. Paul Wilms, the City of Winston-Salem requested that the monitoring frequency for lead be changed from monthly to quarterly for the 001 discharge at the ef- fluent, upstream, and downstream sample sites. Mr. Wilms notified the City in a letter dated December 7, 1989 that our request in this area would be granted. The draft permit, however, still requires monthly monitoring for lead at these three locations. Mr. Dale Overcash March 30, 1990 Page 4 3. Effluent Limitations and Monitoring Requirements for the 002 Discharge The draft permit requires that a discharge via the 002 outfall shall occur only when the USGS gauge located at Highway 64 on the Yadkin River exceeds 5,000 CFS. This limitation is impossible to comply with because the , plant's operators have absolutely no control over the plant dis- �k= charges via the 002 outfall. If the water level in the Yadkin 5 River rises high enough to surcharge the 001 outfall, the plant's ''' � effluent automatically overflows into the 002 outfall leading to Muddy Creek. �� The City objects to this permit requirement because it in- ``(:',ocludes a condition we cannot control. We request the wording in 0 i''this area be deleted from the permit. o'0 The draft permit also requires monthly effluent monitoring at the 002 discharge for Total Nitrogen, Phosphorus, Cadmium, Chro- mium, Copper, Nickel, Lead and Zinc. The City of Winston-Salem requests that monitoring require- ments for all these parameters be deleted from the permit. DEM normally requires monitoring for parameters of this type to assess the long term effects of a discharge o'n a receiving stream. The 002 discharge will have no long term effect on the Muddy Creek be- cause its discharges are infrequent and, even then, occur only during flood conditions. To date, the 002 discharge has been used for a total of ten hours in four years. The City does not feel that the 002 discharge has sufficient occurrences or frequency to warrant monitoring for these param- eters. 4. Bypass In the City's September 15, 1989 permit modification request to Mr. .Wilms, we asked that DEM revise Part II.B.3 of the permit to make the language consistent with the federal bypass regula- tions [40 CFR Part 122.41(m)(4)(A)] Mr. Wilms response of December 7, 1989 to our request in this matter stated that DEM would modify the language in this part to Mr. Dale 0vercash March 30, 1990 Page 5 state the requirements of 40 CFR Part 122.41(m). The draft permit does not contain these language modifications. Inasmuch as we would like to comment on the new language prior to this issue being resolved, DEM should submit a draft of the proposed language to the City for comment. 5. Language in Part III.A.4(e) In the City's September 15, 1989 permit modification request we asked that the language in Part III.A.4(e) of the permit be de- leted or modified. The existing language would prohibit the City from deleting industries from the pretreatment program once they had gone out of business. DEM stated in Mr. Wilm's letter of December 7, 1989 that the language in Part III.A.4(e) will be deleted entirely. This change is not shown in the draft permit. The City requests either the deletion, or alternatively that the provisions in Part III.A.4.e. be amended to read "has been included in the permittee's pretreatment monitoring program submitted in accordance with 15 NCAC 2H.0006 and has not been eliminated in accordance with 15 NCAC 2H.0907(b) and (c)." 6. Terms" Major Contributing Industry" VS. "Significant Industrial User" The City of Winston-Salem asked DEM in the City's permit modification request of September 15, 1989 to either remove the term "Major contributing industry" from Part III.A.1 and III. A.3 or to define the term to distinguish it from the term "Significant Industrial User." The City's request was based on our need to un- derstand which industrial users must be included in the pretreatment program. DEM has indicated that, in an effort to promote clarity, the word major will be deleted. The City feels that clarity would be best served by substituting "SIU" for "major contributing indus- try" in both of these parts. This substitution will not alter the intent of the section. For instance, Part III.A.1 indicates that pollutants in the plant's discharge from industrial contributions may receive NPDES limits as information becomes available. An industry discharging Mr. Dale 0vercash March 30, 1990 Page 6 pollutants which pass through the POTW making it necessary for DEM to impose effluent limitations on these pollutants, would meet the definition of "significant industrial user", although DEM has used the term "major contributing industry." Therefore, the two terms are synonymous in their meanings and one term should be used. Further, Part III.A.3 of the permit states that the Permittee may have to impose more stringent limits on contributing indus- tries than are stipulated in 40 CFR Part 403 to meet the NPDES permit's effluent limits. Part III.4.d, of the permit includes in the definition of "Significant Industrial User" those industries which the permittee has determined to have a potential to ad- versely impact the treatment plant, receiving stream, or to limit the plant's sludge disposal options. Therefore, a contributing industry which received more stringent discharge limitations, as required under Part III.A.3, would automatically become an SIU be- cause the City would have ascertained that there was a need to circumvent an adverse impact on the plant by imposing more strin- gent discharge limits on the industrial user. Again, the terms SIU and major contributing industry are synonymous. The City asks that the last sentence in Part III.A.3 which refers to "major contributing industries" be deleted since the de- letion will not alter the intent of the section. We request that DEHNR also substitute "Significant Industrial User" for major con- tributing industry in all of Parts III.A.1 and III.A.3. 7. Permit Reopener In the City of Winston-Salem's September 15, 1989 request for permit modification, we objected to the broad reopener provisions in the permit. We felt that these provisions, which include parts of Parts II.A.4, II.A.5, III.G and III.H, had the potential effect of subjecting the City to a constant array of changing permit re- quirements necessitating immediate responses which could preclude a coordinated planned response to environmental concerns. DEM has responded that it is not their policy to arbitrarily open permits and that the provisions of 40 CFR 122.62 and NCGS 143-215.1 allow DEM to carry over all reopeners that are in place in the current permit. DEM also maintains that regulations allow the Director to reopen the permit to incorporate new or compli- cated requirements and to incorporate toxic limits. Mr. Dale 0vercash March 30, 1990 Page 7 The City agrees that federal law allows reopeners based on 307-A toxics, but the modification of any other areas of the per- mit is not allowed without the permission of the permittee. Fur- ther, we feel that DEM's response regarding reopeners allowed un- der NCGS 143-215.1 applies only to new and enlarged facilities. In this light, we propose that Part II.H be deleted since toxics reopener provisions are already covered under II.A.5. Part III.G of the permit should also be deleted since it provides for the permit to be reopened to incorporate new effluent limits and/or water quality standards that are not toxics standards es- tablished under Section 307 (a) of the CWA. 8. Authorization of Construction The City of Winston-Salem has requested that the language in Part III.D of the permit be modified to make the requirements of the provisions workable and consistent with federal requirements. As written, Part III.D requires that the City undertake no construction or additions to the facility without written approval from DEHNR. We feel that this requirement is unworkable in that minor modifications must be done routinely to efficiently operate the plant, and neither the City nor DEM has the manpower to pre- pare and process the plans and specifications in a timely manner for the number of changes that would fall under this broad permit provision. DEM has responded to our concerns in this matter by stating that the intent of the provision in Part III.D. is to ensure that all changes at the plant are reviewed and approved. DEM's posi- tion is that there is no need to review a one to one change but moving a component or changing the sizes of components must be ap- proved. We suggest that this provision be limited to those construc- tion projects undertaken to add to the plant's treatment capacity or to those projects intended to change the type of process used in the plant's system. Moreover, the D. C. Court of Appeals has held that EPA lacks the authority to impose a construction ban under the NPDES pro- gram. NRDC VS EPA, 822 f.2d 104 (D.C. Cir. 1987). In view of this, the City asks that wording be inserted into Part III.D stat- ing that this requirement is not enforceable under Federal NPDES procedures. Mr. Dale Overcash March 30, 1990 Page 9 12. Noncompliance Reporting The City of Winston-Salem has requested that DEM delete the provision of Part II.D.6.d of the permit which require that DEHNR be notified by telephone within twenty-four hours of the City be- coming aware of noncompliance with the NPDES permit. While the City has every intention of continuing to notify DEM's regional office when permit exceedances occur, we do not want this function regulated by the permit. DEM has stated that Part II.D.6.d can be deleted and the City requested in a January 11, 1990 letter that the deletion be made. We are unsure if DEM has complied with this request because Part II was not included in the draft permit issued for public comment. 13. Reporting Activities Resulting in the Discharge of Certain Pollutants The City of Winston-Salem has requested that DEM delete all provisions in Part II.D.7. This permit condition is based on the requirements in 40 CFR 122.42(a) which are limited to NPDES per- mits issued to specified industrial dischargers. EPA has deter- mined that these requirements are not appropriate for use in regulating municipal wastewater treatment plants such as the Muddy Creek Plant. DEM has stated that it feels the requirement is appropriate and that it does not have to remove these conditions from the per- mit just because EPA did not include these prov_sions in its mu- nicipal permits. The City of Winston-Salem considers the provision of Part II.D.7 to be more restrictive than 142.42(b) and inconsistent with the provisions of NCGS 143-215(c). NCGS 143-215(c) provides that "effluent standards, limitations and management practices . . shall be no more restrictive than the most nearly: applicable fed- eral effluent standards and limitations and management practices." Mr. Dale Overcash March 30, 1990 Page 8 9. Operation and Maintenance The City of Winston-Salem had requested that DEM modify the language in Part I.B.2 and II.B.1 of the permit to make them con- sistent with the requirements of 40 CFR Part 122. As written, these provisions require that the City operate the treatment plants' facilities at optimum efficiency or as efficiently as pos- sible. DEM has stated that the language in II.B.1 is being modified to coincide with 40 CFR Part 122. DEM did not, however, indicate if a similar modification was to be made to Part I.B.2. 10. Maintenance Scheduling The City of Winston-Salem requested modification of the lan- guage in Part II.D.2 of the permit to allow for a distinction be- tween maintenance events that can be scheduled and those emergency maintenance activities which cannot. The City also requested that the term "non -critical water quality period" be clarified. DEM indicated in their response of December 7, 1989 that they recognized a distinction between scheduleable maintenance and emergency maintenance functions. DEM also indicated that "non -critical water quality periods" were those times when a stream flow greater than 7Q10 coincided with a low flow period through the plant. The City agrees with DEM's response and suggests that the second sentence in the provisions of Part II.D.2 be changed to read as follows: "Any planned, non -emergency maintenance of facilities, which might necessitate unavoidable interruption of operation and possible permit exceedances, shall be scheduled during times when a non-7Q10 stream flow coincides with a period of low flow through the plant." 11. Reporting of Past or Planned Activities In response to the City of Winston-Salem's request for the deletion of Part II.A.4, DEM has agreed to delete the last para- graph in Part II.A.4 and incorporate the language in 40 CFR 122.41(1)(1). Mr. Dale 0vercash March 30, 1990 Page 10 14. Monitoring Frequency The City of Winston-Salem has requested the deletion of Part II.D.4 and Part III.f of the permit, which would give DEM the au- thority to require the City to monitor more frequently or monitor for pollutants not specified in the permit. The City is willing to determine on a case -by -case basis whether increased monitoring requirements are justified, but we object to the modification of the permit without the City's con- sent and the absence of provisions to protect the City's right to due process. DEM has responded that NCGS 143-215.1(b)(3) gives DEM the right to change the monitoring frequency upon sixty (60) days no- tice. The City continues to maintain that DEM does not have this authority in that the provisions of NCGS 143-215.1(b)(3) pertain to new or enlarged facilities. The Muddy Creek Plant does not fall into either of these two categories. Inasmuch as this area of the permit remains unresolved, the City of Winston-Salem would welcome an opportunity to discuss the matter further with DEM. 15. Monitoring of GC/MS Peaks Part I.J.2 of the permit requires that the ten largest GS/MS peaks in each organic chemical analytical fraction be quantified and identified. The City of Winston-Salem requested a modifica- tion to this permit provision to limit the requirement to those GC/MS peaks in each fraction, not to exceed ten, which can be identified with a confidence level of at least ninety percent (90%). DEM has stated that it is evaluating the language and will make some changes in the permit. Since the provisions of this part are not included in the draft permit issued for public comment, the City is unsure as to their status. The City does reserve the right to comment on any language changes DEM proposes for Part I.J.2 and we would welcome an opportunity to discuss the matter further, should DEM so de- sire. Mr. Dale Overcash March 30, 1990 Page 11 16. Pretreatment Program Resubmission Part III.A.4 of the draft permit requires the City to submit a Pretreatment Program to DEHNR for approval prior to accepting discharges from any new SIUs. This provision is inappropriate; Winston-Salem already has an approved pretreatment program. Fur- ther, federal and state pretreatment program requirements do not require a new pretreatment program to be developed every time a new SIU begins discharging. This permit requirement is unneces- sary. The City of Winston-Salem's permit modification request of September 15, 1989 petitioned DEHNR to modify the mechanism by which the new SIU's could be added to the City's pretreatment pro- gram. DEHNR responded with a proposal to require the City to submit a modified description of the pretreatment program for approval, rather than an entire program. The City of Winston-Salem is concerned about the possible ramifications of the use of the term "modified program descrip- tion" in the proposed language for Part III.A.4. Section III.B.12 of the permit provides that pretreatment program modifications shall be considered as permit modifications, thus opening every permit provision for possible change each time a new SIU is per- mitted. The City feels that the addition of SIU's to the pretreatment program is not a modification as much as it is a re- porting function. The City requests that the second sentence in the first para- graph of Section III.A.4 be modified to read as follows: "Prior to accepting wastewater from any new significant user, the permittee shall submit a permit and synopsis for the SIU to DEHNR in accordance with Section 15 NCAC 2H.0906 of the North Carolina Administrative Code. The Permittee shall not be required to prohibit discharges from existing industrial users, being reclassified as a significant user, while DEHNR is reviewing the permit." 17. Submission of Information Requested by DEHNR The City of Wlnston-Salem has requested that the provisions of Part III.B.6(f) of the permit be deleted on the, grounds that ? t'. Mr. Dale Overcash March 30, 1990 Page 12 the language in this provision is not contained in NCAC 2H .0908 and it could be interpreted as giving DEHNR the authority to re- quire reporting activities well in excess of that provided for by the regulations. We suggest that the language in Part III.B.6(f) be changed to read as follows: "upon request, other information which is necessary to deter- mine compliance with the pretreatment program and required by the provisions under NCAC 2H.0908." 18. Change in Definition of Significant Industrial User In its permit modification request of September 15, 1989, the City of Winston-Salem requested that DEHNR delete the last sen- tence under Part III.A.4. The provisions of this part would auto- matically incorporate into the permit the changes in the defini- tion of SIU occurring as a result of promulgations in response to Section 307 of the CWA or revisions to 15 NCAC 2H .0903. We felt that this provision deprived us of our rights to due process. DEHNR notified us in a December 7, 1989 letter that this pro- vision would be deleted. This change was not made in the text of the draft permit issued for public comment and it should be ad- dressed by DEM. 19. Modifications to North Carolina's Pretreatment Program The City of Winston-Salem is concerned that DEHNR may have made significant changes in the State's pretreatment program with- out formal EPA approval, as required by 40 CFR Part 123.62. Inasmuch as the City objects to NPDES permit requirements be- ing imposed based on State regulations which have not been for- mally approved by EPA, the City requests that a statement be added to Part III.B of the permit to read as follows: "Pretreatment program requirements incorporated into the per- mit which have not been formally approved by EPA as per 40 CFR 123 are not deemed to be federal requirements and are not federally enforceable." The City of Winston-Salem is dedicated to the protection of Mr. Dale Overcash March 30, 1990 Page 13 water quality in North Carolina. We look forward to a cooperative effort with DEHNR in this regard. All of our comments are being made with a constructive intent and we would welcome the opportu- nity to discuss these issues in more detail. If you have any questions regarding these comments or the supporting attachment, please feel free to call Stan Webb at 1/919-784-4700. Your cooperation in this matter is greatly ap- preciated. pc: rl r Crystal Couch Stan Webb Dr. George Everett Larry Coble Attachment(s) SW/lt Sincerely, Tom Gri•f.fin P. E. UTILITIES DIVISION Utilities Superintendent Page 2 While we have no data which indicates conclusively that this is occurring, we would like to point out that 44% of the samples failing the test based on reproduction had reproductive rates which were equal to or higher than the average control and much higher than the level of reproduction known to occur naturally. Our findings suggest that it would be prudent to conclusively identify the factor or factors which cause "reverse toxicity" and modify the test procedures to compensate for their effects. II. The test protocol does not utilize an effective control in that test organisms are subjected to non -toxicant related stresses to which control organisms are not exposed. Under the test protocol, organisms slated to serve as con- trols are transferred into the same solution in which they were cultured, and those individuals comprising the test group are sud- denly placed into a solution which may have altogether different, although normal, levels of water hardness, pH, conductivity, and osmotic pressures. Therefore, test organisms undergo stresses associated with adapting to a new environment which control organisms do not. It is entirely conceivable that these sub lethal stresses could cause reproductive impairment. The test would therefore give a false indication of toxicity. The City of Winston-Salem feels that this portion of the pro- tocol serves to defeat the purpose of the control - that being to equalize the effects of every parameter on the test result except toxicants. The test protocol should be modified to ensure that the back- ground concentrations of all normal water qualities are ap- proximately equal in both the test and control solutions. III. The pass/fail mini -chronic procedure does not exhibit the degree of accuracy in measuring reproductive impairment that is necessary to warrant its use as an enforcement tool. During the City of Winston-Salem's review of test data ob- tained from the pass/fail mini chronic procedure, data were ob- tained which indicated the test protocol does not accurately mea- sure reproductive impairment. Page 3 Six sets of replicates comparing control water to itself were statistically analyzed according to the provisions of Appendix H in EPA 600/4-89/001. The critical values used to measure statis- tical fit were chosen to reflect a 99% confidence factor, as re- quired by DEM. Five of the six replicates predicted significant reproductive impairment where there could not possibly have been any difference in the water samples. This data is detailed in Table III-1. Table III-1: Results of Control Replicate Analysis Conducted on N. C. Mini -Chronic Bioassay Procedure Sample Reproduction X 1 Test I.D. I-11 Date 5-9-89 1 2 3 4 5 6 7 8' 9 10 11 12Aepo.Result 24 10 14 13 13, 23 12 23. 23 9 231 8 16.2 Fail I-10 5-9-89 28 22 27 34 24 31 31 30 26 ' 16 20 25 26.2 I-17 10-18-89 24 27 30,27 24,33 26 27 29 , 31 26 14, l3 26.5 31.; Fail I-18 10-18-89 _ 27 34- 30 31131 31 32 33! 30 32 30 I-17 _ 10-18-89 24 27 30 27 24 33 26 27 29 31 26 14, 38 26.5 35.2 Fail 1-19 10-18-89 38 33 41 32 37 33 31 41 29 37 32 I-18 10-18-89 27 34 30 31_31 31 32 33 30 32 30 33 31.2 Fail 1-19 10-18-89 38 33 41 32 37 33 31 41 29 37 32 38 35.2 I-23 12-27-89 31 29 26 29 27 30 36 5 _27 _27 ,34 35 28 Pass 1-24 12-27-89 36 34 31 `27 31� 33 30 35 31 32.4 I-26 1-10-90 36 37 28 38 ,38 ,37_30 29 35 3.6 36 _34 37 36 34.2 Fail 1-25 1-10-90 41 41 ,34. 41 , 35 43 43 39 38 40 44 37 45 40.6 To further evaluate the possibility of a sample being falsely declared toxic on the basis of reproductive impairment, the repro- ductive data obtained on all samples failing the test non -acutely were compared against every control group in the data set. Our goal in doing this was to determine the percentage of instances when a reproductive based test failure would pass, when compared to a large population of individual controls. The summary of these comparisons are shown in Table III.2. Detailed data are included as well in Appendix B. Page 4 Table III-2: Summary of Statistical Comparisons Between Repro- ductive Impairment Test Failures and a Large Po- pulation of Control Groups. Parameter # of Comparisons 66 # of Test "Passes" 49 t Test Passes with Reproductive Impairment Test Failure Designation A B C D E F G H I 66 66 66 66 66 66 66 66 32 57 21 23 15 63 64 11 "Reverse Toxicity" 21 0 30 0 0 0 34 53 0 % of Time Passing 74% 48% 86% 32% 35% 23% 95% 97% 17% of Time Reverse Toxicity Occurred 32% 0% 45% 0% 0% 0% 52% 80% 0% There were nine test reports in the total data set which failed due to reproductive impairment. Of these nine, four had reproductive rates in the test group which was statistically iden- tical to or superior to at least seventy-four percent of the sixty-six control groups used in the comparison. We feel that this translates into at least 44% probability of a sample being declared toxic based on reproduction when it probably is not. The data on replicate control groups indicate this probability may be much higher. IV. DEM's Policy of Defining Permit Noncompliance as two Succes- sive Failures of the Pass/Fail Procedure is not Consistent with EPA Recommendations as to how Bioassay Data should be Utilized as an Enforcement Tool. EPA's "Policy for the Development of Water Quality -Based Per- mit Limitations for Toxic Pollutants", parts of which are refer- enced in EPA 600/4-89/001; March, 1989, states that "Effluent tox- icity data, in conjunction with other data, can be used to establish control priorities, and set permit limitations to achieve those standards." The qualifier statement, "in conjunction with other data", indicates that EPA does not intend for the results of a bioassay test to stand alone in determining a facilities compliance status with whole effluent toxicity limitations. EPA also recommends that bioassays intended to be used in conjunction with other data to demonstrate noncompliance should be conducted so as to determine the actual NOEC of the sample and Page 5 ensure that the response of the test organisms to increasing con- centrations of toxicants is linear. This requires that a series of sample dilutions be evaluated simultaneously, and the mini -chronic procedure does not do this. DEM's current policy of assessing noncompliance with effluent toxicity limitations is based solely on the occurrence of two con- secutive test failures. There is no consideration given to other pertinent data, nor are there provisions in the mini -chronic test protocol to evaluate the dose response. In evaluating the data presented in this document, the City of Winston-Salem was convinced that the pass/fail mini -chronic bioassay test is not suitable for use in determining compliance. It may be useful in monitoring to determine if a facility's dis- charge needs further evaluation, but even the use of the test for this purpose is questionable if test failures in one or two samples are considered to be significant. The City of Winston-Salem feels that the pass/fail mini -chronic procedure is accurate only when its predicts toxicity associated with mortality. In this respect, the procedure is no more accurate than the "Acute Toxicity Test." It Master list of all data evaluated A la =' Appendix A PRT = PASS WITH REVERSE TOXICITY = FAI ;AMPLE I.D_ UMBER TEST DATE REPRODUCTION AVG. 'EPROD.UMHOS COND. HARD NESS MG/L SITE CODE 1 2 3 4 5 6 7 8 9 10 11 12 1-1 .. 26 21 34 32 34 28 27 22 15 36 29 29 27.8 260 28.7 IWC= (93%) 28 30 30 21 28 25 19 23 29 28 16 15 P 24.3 550 1-2 7-15-88 45 41 36 42 34 46 40 37 31 29 42 39 38.5 228 33.5 2 (96%) 26 30 26 31 36 31 27 32 34 29 41 39 F 31.8 305 1-3 8-21-88 27 11 16 19 15 19 10 17 11 17 11 19 16.0 490 49 3 (93%) , 22 14 0 14 1 6 15 15 12 13 F 9.8 335 1-4 10-3-88 24 29 30 29 26 28 29 32 29 29 29 28 28.5 200 38 4 (5%) 38 35 34 38 41 39 31 40 38 35 38 42 PRT 37.4 420 1-5 10-17-88 31 19 30 17 34 20 20 36 34 18 31 28 26.5 200 38 5 (62%) 47 46 43 44 47 39 43 41 41 38 47 42 PRT 43.2 390 1-6 10-25-88 17 8 16 20 16 12 16 12 10 21 18 15 15.1 186 49.9 1 (93%) 42 47 40 45 49 42 34 37 27 47 36 47 PRT 41.1 609 1-7 3-13-89 27 10 13 35 19 34 12 37 20 36 31 39 26.1 156 43.5 5 (62%) 39 36 34 38 34 32 41 43 36 43 39 47 PRT 38.5 400 1-8 4-24-89 31 32 32 31 35 31 M 26 29 24 23 21 28.6 182 43.5 6 (36%) 36 33 32 38 25 28 30 34 33 30 30 31 PRT 31.7 290 1-9 5-11-89 8 24 30 24 9 18 25 13 10 23 4 21 17.4 178 37.5 5 (62%) 47 56 48 52 47 43 49 50 46 50 43 48 PRT 48.2 350 Appendix A Page 2 of 11 SAMPLE I.D. NUMBER TEST DATE REPRODUCTION A AVG. COND. HARD NESS MG/L SITE CODE 1 ?_ 3 4 5 6 7 8 9 10 11 12 'EPROD.UMHOS 1-10 5-9-89 28 22 27 34 24 31 31 32 26 16 20 25 26.2 178 37.5 21B (100%) 28 32 24 32 29 33 31 45 25 26 25 29 PRT 29.9 123 1-11 5-9-89 24 10 14 13 13 23 12 23 23 9 23 8 16.2 178 37.5 21A (100%) 35 42 28 32 35 34 34 38 38 31 42 31 PRT 35.0 128 1-13 7-17-89 29 31 24 29 34 27 33 31 31 32 29 22 29.3 158 36.9 6 31 39 22 13 46 30 26 24 23 47 27 38 P 30.5 191 (36%) 1-14 8-7-89 45 45 47 40 46 43 41 40 44 46 47 42 43.8 151 36.9 5 (62%) 25 43 51 46 42 29d 55 47 23d 37 15 57 P 39.2 432 28.1 163 37 6 24 28 28 28 30 27 31 , 25 29 30 30 27 1-16 10-2-89 (36%) 31 30 35 32 19 41 30 M 35 31 34 33 PRT 31.9 167 1-17 10-18-89 24 27 30 27 24 33 26 27 29 31 26 14 26.5 158 40 7 (98.7) 0d 0d 0d Od Od 0d Od Od 0d Od Od Od F 0 428 1-18 10-18-89 27 34 30 31 31 31 32 33 30 32 30 33 31.2 158 40 7 (98.7) 0 0.i 0d 0 0 0d Od 0d Od 0 Od 0 d F 0 428 1-19 10-18-89 38 33 41 32 37 33 31 41 29 37 32 38 35.2 158 40 4 (4%) 34 Od 21 0 d Od Od 23 27 30 15 15 19 F 15.3 411 1-20 11-8-89 35 28 32 33 33 37 22 29 26 25 29 36 30.4 172 37 5 (62%) 36 24 11 0 d 25 24 1 ,; 0 d 30 0 d 12 22 F 15.3 435 Appendix A Page 3 of 11 ;AMPLE I.D. 1UMBER TEST DATE REPRODUCTION AVG. REPROD_UMHOS COND. HARD NESS MG/L SITE CODE 1 2 3 4 5 6 7 8 9 10 11 12 1-21 11-29-89 29 30 18 32 32 32 33 35 28 32 21 30 29.3 160 40 5 (56%) 35 17 28 24 35 30 19 30 27 9 33 26 P 26.1 343 1-22 12-13-89 42 29 11 19 40 34 38 36 41 38 18 39 32.1 163 44 4 (4%) 32 22 36 35 34 31 29 32 38 38 21 35 P 31.9 367 1-23 12-27-89 31 29 26 29 27 30 3.6 5 27 27 34 35 28.0 161 44 5 (56%) 37 31 40 44 35 34 38 40 32 39 40 34 PRT 37.0 408 1-24 12-27-89 36 34 31 27 31 33 37 30 30 35 34 31 32.4 161 44 4 (4%) 33 36 43 34 38 36 34 43 38 32 37 38 PRT 36.8 363 I-26 1-10-90 41 41 41 35 43 43 39 38 40 44 37 45 40.6 155 44 4 (4%) 41 42 38 40 38 38 36 30 36 38 38 34 F 37.4 448 1-25 1-10-90 36 37 34 28 28 38 29 35 36 36 37 36 34.2 155 44 6 (36%) 31 2 32 39 41 45 35 38 39 42 38 43 P 35.4 166 1-27 7-19-88 30 16 23 30 22 29 36 30 33 24 31 30 27.8 220 29.8 6 (36%) 35 31 32 33 23 27 19 23 27 21 33 30 P 27.8 230 1-28 6-1-88 27 25 26 33 19 20 25 25 31 28 29 31 26.6 140 11.1 8 (37%) d0 0 d d0 d 0 d0 0 d d0 d 0 d0 d0 d0 d0 F 0 750 1-29 2-20-89 25 39 35 35 42 34 37 30 36 38 43 40 36.2 133 43.5 9 (1.4%) 33 38 35 35 35 38 35 39 36 7 43 40 P 34.5 722 Appendix T. Page 4 of 11 'AMPLE I.D. 4UMBER TEST DATE REPRODUCTION AVG. 'EPROD.UMHOS COND. HARD NESS MG/L SITE CODE . 2 3 4 5 6 7 8 9 10 11 12 1-30 3-6-89 14 12 23 18 20 3 15 19 12 19 20 8 15.3 168 43.5 9 (1.4%) 10 20 9 11 10 9 4d 12d 6 9d1 d 6 F 8.9 666 1-31 7-10-89 22 33 33 26 43 34 30 37 30 30 31 29 31.5 170 36.9 10 (84%) °d d7 10d 28d 14d 14d . 10d 20d 11a 15d 19d F 13.5 422 1-32 5-8-89 8 24 30 24 9 18 25 13 10 23 4 21 17.4 178 37.5 5 (62%) 55 50 29 36 43 47 51 53 49 46 36 50 PRT 45.4 336 III-1 10-23-89 15 15 23 27 3d 23 23 4 d 25 23 30 19.8 130 30 11 (72%) O d d0 d9 4 d Od '. d6 d II O d 5d 6 F 2.8 460 IV-1 10-3-88 27 24 22 21 22 20 18 20 20 23 24 18 21.58 84 45 4 (5%) 12 20 19 13 M 10 11 6 7 8 6 8 F 10.91 430 IV-2 11-15-88 17 25 19 13 22 31 28 17 24 23 18 18 21.25 80 48 1 (93%) 28 26 16 24 16 26 26 24 29 34 26 25 PRT 25.0 630 IV-3 9-11-89 30 35 31 37 39 31 40 35 37 30 34 33 34.3 70 40 7 (99%) 28 30 36 29 26 33 33 27 28 32 26 32 F 30.0 390 IV-4 10-12-89 34 32 33 39 33 36 33 29 39 33 36 36 34.4 60 40 12 d-H20 7 11 9 7 6 2 d5 4 d 0 3 F 4.5 20 • Appendix A Page 5 of 11 SAMPLE I.D. UMBER TEST DATE REPRODUCTION HARDt 1 2 3 4 5 6 7 8 9 10 11 12 'EPROD.UMHOS AVG. COND. NESS MG/L SITE CODE IV-5 10-18-89 36 32 32 34 41 32 38 34 36 39 33 38 35.4 60 40 7 (99%) 24 20 21 30 33 29 25 27 21 25 28 25 F 25.7 355 IV-6 10-25-89 29 33 30 26 32 30 37 33 28 35 35 29 31.4 50 40 13 (99%) 36 39 30 27 33 31 30 37 33 36 39 44 P 34.6 345 IV-7 4-24-89 18 13 19 13 19 14 16 14 21 15 16 15 16.1 80 40 14 15 24 21 .26 22 20 18 18 21 20 20 14 PRT 19.9 305 (37%) IV-8 4-24-89 18 13 19 13 19 14 16 14 21 15 16 15 16.1 80 40 8 (37%) Oa 0 Od 11 . Od Od Od Od F 0 910 36 32 32 34 41 32 38 34 36 39 33 38 35.4 60 40 15 IV-9 10-18-89 (99%) 43 34 29 37 36 31 30 35 34 42 40 40 P 35.9 340 IV-10 10-25-89 29 33 30 26 32 30 37 33 28 35 35 29 31.4 50 40 11 (72%) 23 29 17 32 26 14 12 0 11 16 20 0 F 16.7 345 IV-11 10-25-89 29 33 30 26 32 30 37 33 28 35 35 29 31.4 50 40 16 (99%) 29 33 28 29' 31 30 29 34 26 43 20 14 P 28.8 360 IV-13 5-29-89 16 16 20 15 16 15 17 16 23 23 24 29 19.2 100 40 10 (84%) 27 26 20 23 26 26 26 20 22 22 22 25 PRT 23.75 370 Appendix A Page 6 oi: 11 AMPLE I.D. 1UMBER TEST DATE REPRODUCTION AVG. REPROD . COND. UMHOS HARD NESS MG/L SITE CODE 1 2 3 4 5 6 7 8 9 10 11 12 IV-14 5-30-89 16 16 20 15 16 15 17 16 23 23 24 29 19.2 100 40 8 (37%) 8 17 6 6 21 19 17 16 10 10 8 7 F .. 12.1 970 IV-15 6-19-89 33 30 21 29 28 34 23 38 26 36 33 24 29.6 90 40 10 (84%) 0 19 0 19 0 d 28 Od 0 0 0 0 20 F 7.2 325 IV-16 6-19-89 33 30 21 29 28 34 23 38 26 36 33 24 29.6 90 40 8 (37%) 30 0 23 23 7 0 23 0 20 26 0 0 F 12.7 795 IV-17 6-26-89 20 31 27 20 30 29 34 32 31 35 28 37 29.5 70 40 9 (1.4%) 24 28 30 30 19 28 28 29 14 30 38 33 P 27.6 650 IV-18 6-26-89 20 31 27 20 30 29 34 32 31 35 28 37 29.5 70 40 10 (84%) 27 25 28 27 24 22 16 24 27 31 22 29 P 25.2 490 V-1 10-23-89 27 24 37 25 28 35 37 24 39 29 34 33 31.0 162 40 13 (99%) 36 28 32 20 24 33 29 27 30 35 33 20 P 28.9 487 V-2 12-6-89 26 22 22 20 17 18 21 22 20 18 18 15 19.9 116 46 11 (72%) 25 23 26 27 29 28 30 29 16 33 29 23 P RT 26.5 475 V-3 12-13-89 18 26 23 32 23 22 28 32 27 18 26 23 24.8 116 46 22A A Page 7 of 11 SAMPLE I.D. UMBER TEST DATE REPRODUCTION AVG. COND. HARD NESS MG/L SITE CODE 1 2 3 4 5 6 7 8 9 10 11 12 'EPROD.UMHOS (99%) 6 6 16 20 4 18 24 25 14 5 0 I F • 138 V-4 8-9-88 16 16 18 15 15 17 16 16 17 14 15 18 16 250 148 3 93%) 12 16 16 16 16 15 13 17 18 17 18 16 P 15.8 213 V-5 8-23-88 16 16 17 17 16 15 18 17 15 15 16 16 16.2 157 --- 3 (93%) 14 15 16 16 16 21 14 16 16 14 16 16 P 15.8 416_ V-6 10-23-89 27 24 37 25 28 35 37 24 39 29 34 33 31.0 162 40 16 (99%) 26 30 32 27 28 26 21 29 34 33 23 29 P 28.2 564 V-7 12-6-89 26 22 22 20 17 18 21 22 20 18 18 15 19.9 116 46 13 (99%) 20 15 17 16 0 d 14 . lld 26 13 11 14 8d F 13.8 502 r V-8 12-6-89 26 22 22 20 17 18 21 22 20 18 18 15 19.9 116 46 17A (100%) 10 d 0 15 12 15 27 26 11 14 20 9 23 P 15.2 203 V-9 12-6-89 26 22 22 20 17 18 21 22 20 18 18 15 19.9 116 46 16 (99%) 20 29 26 24 27 25 23 24 26 15 29 23 PRT 24.3 511 V-10 12-6-89 26 22 22 20' 17 18 21 22 20 18 18 15 19.9 116 46 18A (100%) 25 11 13 20 22 23 24 13 16 16 23 15 18.4 190 V-11 12-6-89 26 22 22 20 17 18 21 22 20 18 18 15 19.9 116 46 19A (100%) 4 0 1 11, 3 0 13 0 16 9 0 16 6.1 185 V-12 12-13-89 18 26 23 32 23 22 28 32 27 18 26 23 24.8 116 46 7 T ,1n 'r9 1 X A r ;AMPLE I.D. IUMBER TEST DATE REPRODUCTION AVG. COND. HARD-t' NESS MG/L SITE CODE 1 2 3 4 5 6 7 8 9 10 11 12 'EPROD.UMHOS 22 20 24 22 22 19 23 14 26 23 14 26 P 21.3 425 (99%) V-13 12-13-89 18 26 23 32 23 22 28 32 27 18 26 23 24.8 116 46 20A (99%) 27 23 20 17 25 26 21 13 14 30 17 28 P 21.8 121 V-14 12-13-89 18 26 23 32 23 22 28 32 27 18 26 23 24.8 116 46 15 (99%) 14 18 14 12 23 9 22 16 12 21 12 2.1 F 16.2 415 VI-1 6-7-88 22 20 20 19 15 22 20 19 20 21 21 19 20.0 115 45 5 P 18.42 280 62% 18 15 15 20 20 23 17 16 19 15 22 21 VI-2 3-14-89 21 24 27 13 16 17 18 25 31 32 28 19 22.6 140 39 6 (36%) 22 14 5 18 32 7 29 17 14 22 10 22 P 17.7 382 VI-3 10-16-89 24 0 d 23 32 29 22 49 0 d 27 36 33 20 24.6 168 45 15 (99%) 23 35 32 27 32 31 43 24 28 26 32 24 PRT 29.8 478 VI-4 11-8-89 35 33 42 33 34 33 1 a 30 30 38 29 31 30.8 168 45 9 (1.4%) 31 31 27 28 25 26 29 26 30 34 30 34 P 29.3 488 VI-5 9-4-89 27 32 29 29 24 24 26 28 26 12 19 31 25.6 168 41 9 (1.4%) 26 21 20 27 23 13 23 22 27 13 28 19 P 21.8 762 VI-6 8-14-89 26 5 32 17 24 29 26 20 23 25 27 24 23.2 168 40 9 • Appendix Page 9 of 11 AMPLE I.D. UMBER TEST DATE REPRODUCTION AVG. COND. HARDt NESS MG/L SITE CODE 1 2 3 4 5 6 7 8 9 10 11 12 REPROD.UMHOS 27 •29 20 21 29 24 27 27 24 21 27 24 P 25.0 661 (1.4%) VI-7 8-7-89 23 19 19 20 24 22 24 21 18 27 23 22 21.8 168 35 10 (84%) 24 27 16 30 18 27 24 28 28 24 18 24 P 24.0 321 VI-8 9-11-89 9 d 22 19 21 23 27 22 19 28 22 26 29 22.25 168 38 10 (84%) Od Od O a O d Od O d. Od 0 d Od 0 d Od Od F 0 402 22 27 31 27 21 20 19 32 24 26 18 24 24.3 168 45 10 VI-9 10-9-89 (84%) Od 18 0 c Od 26 0 di Od Od 3 22 21 22 F 9.3 328 V1-10 11-29-89 24 13 18 14 8 16 17 20 15 15 23 14 16.4 168 48 10 14 29 15 18 29 29 32 16 17 15 13 17 PRT 20.3 345 (84%) VI-11 7-10-89 16 7 25 29 25 24 20 23 21 14 10 23 19.75 114 42 14 (37%) 14 23 22 22 28 19 16 19 21 22 21 23_ P 20.83 307 VI-12 10-16-89 24 0 d 23 32 29 22 49 0 d 27 36 33 20 24.6 168 45 14 (37%) 41 24 41 35 33 36 38 22 43 45 35 40 PRT 36.1 243 VI-13 9-11-89 9d 22 19 21 23 27 22 19 28 22 26 29 22.25 168 38 8 (37%) 2 1 d 5d 7d 6 d ld 6d 3d 7 d 3d 3a 1 d F 3.8 612 V1-14 7-10-89 16 7 25 29 25 24 20 23 21 14 10 23 19.75 114 42 8 (37%) 21 17 19 16 22 16 18 19 17 21 20 18 18.67 696 VI-15 10-9-89 22 27 31 27 21 20 19 32 24 26 18 24 24.3 168 45 8 ;AMPLE I.D. DUMBER REPRODUCTION COND. HARDt NESS MG/L SITE CODE TEST DATE 1 2 3 4 5 6 7 8 9 10 11 12 'EPROD.UMHOS AVG. (37%) 26 20 23 26 21 25 29 20 14 21 23 15 P 21.9. 652 I11-1 11-1-89 13 16 16 14 17 15 18 15 18 20 16 15 16.08 155 56.8 8 (37%) 18 15 18 17 28 32 28 34 16 19 20 13 PRT 21.50 462 111-2 11-2-89 15d 17d 21 19 21 15 18 17 17 11 16 15 16.83 160 56.8 8 (37%) 30 26 24 25 26 21 25 27 20 18 27 13 PRT 23.5 496 VII-3 11-30-89 30 24 14 19 15 21 32 16 25 4 30 27 21.42 138 50.8 8 (37%) 22 26 22 16 21 20 23 21 20 22 21 14 P 20.67 510 V-15 1-10-90 23 22 13 15 20 17 17 21 15 13 16 16 17.3 r 141 43 1 18A 99% 21 3 19 16 18 10 14 21 13 8 22 22 P 14.8 216 V-16 1-10-90 23 22 13 15 20 17 17 21 15 13 16 16 17.3 141 43 17A 99% 25 16 19 28 13 11 18 12 22 15 11 14 P 17.0 166 V-17 1-10-90 23 22 13 15 20 17 17 21 15 13 16 16 17.3 141 43 19A 99% 25 16 19 28' 13 11 18 12 22 15 11 14 P 15.8 106 V-18 1-10-90 23 22 13 15 20 17 17 21 15 13 16 16 17.3 141 43 13 99% 18 12 III13 II 4,'d12 F 10.4 476 V-19 1-10-90 23 22 13 15 20 17 17 21 15 13 16 16 17.3 141 43 11 72% 13 20 16 13 23 23 19 17 24 23 23 21 P 19.6 437 1 J )e1LLt J_ 1-1. SAMPLE I.D. NUMBER TEST DATE REPRODUCTION AVG. COND. HARD NESS SITE MG/L CODE 1 2 3 4 5 6 7 8 9 10 11 12 REPROD.UMHOS 23 22 13 15 20 17 17 21 15 13 16 16 17.3 141 43 16 V-20 1-10-90 99% 20 13 11 11 19 14 16 16 17 12 24 10 P 15.3 486.._ 21.6 141 43 15 V-21 1-17-90 21 15 20 23 25 23 23 24 24 19 22 20 99% 16 27 21 14 21 11 14 18 9 19 21 19 P 17.5 621 V-22 1-17-90 21 15 20 23 25 23 23 24 24 19 22 20 21.6 141 43 7 lld 7d11d14 11 14 15d 7 14 9 13d14 F 11.7 - 457 ■99% V-23 1-17-90 21 15 20 23 25 23 23 24 24 19 22 20 21.6 141 43 22A 99% 30 25 33 24 20 25 27 Od 28 Og 22 25 21.6 214 V-24 1-17-90 21 15 20 23 25 23 23 ` 24 24 19 22 20 21.6 141 43 20A 16 21 28 27 15 4 25 31 30 21 21 28 P 22.3 150 ■99% ■ ■ ■ Appendix 13 Page 1 of 5 Result of Statistical Comparisons Between Reproductive Failures and Total Control Population Control I. D. # of Test Groups Failing on Reproduction I. D. Number A B C D E F G a. 1 I-1 P T P T T T P PRT T I-2 T T T T T T T P T I-3 PRT P PRT P P P PRT PRT P I-4 P T P T• T T P PRT T I-5 P T P T T T P PRT T I-- 6 PRT P PRT • P P P PRT PRT P I-7 P P P 1 T T T P PRT T I-8 P P P. T T T P PRT T I- 9 PRT P PRT P P P PRT PRT P I-10 P T P T T T PRT PRT T I-11 PRT P PRT P P P PRT PRT P • I-13 P T P T T T P• PRT T I-14 T T T T ' T T T T T I-16 P PRT I-17 PRT PRT I-18 PRT KEY P = PASS PRT = PASS WITH REVERSE TOXICITY T = TOXIC I-19 • Appendix 13 Page 2 of 5 Result of Statistical Comparisons Between Reproductive Failures and Total Control Population Control I. D. # of Test Groups Failing on Reproduction I. D. Number A B C D E F G H I I-20 T T P T T T P PRT T I-21 P T P T T T P PRT T I-22 P T P T T T P P T I-23 P T P. T T T P P T I-24 T T P T T T P PRT T III-1 P P PRT P P P PRT PRT T IV-1 PRT P PRT T T T •PRT PRT 1 T IV-2 P P PRT T P T PRT PRT T IV-3 T T T T T T P P T IV-4 T T T T T T P P T IV-5 T T T T T T P P T IV-6 T T P T T T P PRT • T V-1 PRT V-2 PRT PRT PRT PRT V-3 PRT PRT PRT VI-1 PRT •PRT PRT PRT VI-2 PRT PRT PRT • Pppenaix b Page 3 of 5 Result of Statistical Comparisons Between Reproductive Failures and Total Control Population Control I. D. # of Test Groups Failing on Reproduction I. D. Number A B C D E F G H I VI-3 P P P P P P P PRT T I-25 T T T T T T P P T I-26 T T T T T T T T T 1-27 P T P T T T P PRT T • 1-28 P T P T T T PRT PRT T I-29 T T T T T T P P T I-30 PRT P PRT P P P PRT PRT P 1-31 T T P T T T P PRT T 1-32 PRT P PRT P P P PRT PRT P IV-7 PRT P PRT P P P PRT PRT P IV-13 PRT P PRT P P T PRT PRT P IV-15 P T P T T T P PRT T IV-17 P T P T T T' P PRT T V-4 PRT P PRT P P P PRT PRT P V-5 PRT P • PRT P P P PRT PRT T V-6 T T P T T T P PRT T V-7 PRT P PRT P P T PRT PRT T Appendix B Page 4 of 5 Result of Statistical Comparisons Between Reproductive Failures and Total Control Population Control I. D. # of Test Groups Failing on Reproduction I. D.A Number B C D E F G H I V-12 P T PRT T T T PRT PRT T VI-4 T T P T T T P p T VI-5 P T P T T T PRT PRT T VI-6 P P PRT T T T PRT PRT T 'VI-7 PRT P PRT T T T PRT PRT T VI-8 P P PRT T T T PRT PRT T VI-9 P P PRT T T T PRT PRT T VI-10 PRT P PRT P P P PRT PRT P VI-ll PRT P PRT P P T PRT PRT T VI-12 P P P P P P P P T VII-1 PRT P PRT P P P PRT PRT P VII-2 PRT P PRT P P - P PRT PRT T VII-3 P P PRT P P T PRT PRT T V-15 PRT P PRT P P T PRT PRT T V-21 PRT P PRT T T T PRT PRT T Appendix B Page 5 of 5 Summary # Control Comparisons A B C D EFGH I 66 66 66 66 66 66 66 66 66 # Passes 49 32 57 21 23 15 63 64 11 # Pases with Reversible Toxicity 21 0 30 0 0 0 34 53 0 % Passes 74% 48% 86% 32% 35% 23% 95% 97% 17% % Samples with Reverse Toxicity 32% 0% 45% 0% 0% 0% 52% 80% 0% Doubt that test samples A, C, G, H are really fails. �iple D. 1 I -I I-2 I-3 I-4 I-5 I-6 I-7 1-8 --9 10 -11 -13 - 17 -19 :-20 .-21 -22 :-23 :-24 I-1 :II-1 .V-1 :Y-2 :V-3 (V-4 :Y-6 Y-1 V-2 V-3 /1-1 'I-2 /I-3 26 06/14/88 O111S188 08/21/88 10/03/89 IOIlll88 1012SI88 03/13/89 04/28/89 OSIO (89 05/09/89 0810 IB 101�1189 10/11/89 10/18/89 11/06/89 IU27189 12 1J 89 12�2T 89 12/27/89 10/17/88 10123J89 1t115188 Oj12189 10/16/89 10/23/89 12f06189 06108/B8 1 0116�89 01108 j90 ' Control Group Reproduction F Test Test --- Test A vs Date __�1 2 3 41 5 61 l 8 9:10:11:12: Control 26 21 34 32 34 28:27 22 I S 36,29 29, .2.42 4541 36423446:403731 29:4239 1.80 271116 19 15 19:10 17 11 17:11 19 1,54 242930292628:29293229:2928 3.97 31 19 30 11 34 20:20 36 34 18:31 28 3.29 17 8 16 20 16 12 16 12 10 21:18 15 1.01 27 10 13 35 19 34 12 37 20 36131 39 1.42 31 32 32 31 3531 26 29 24:23 21 8243024 91825 1310231 421 4.46 282221342431 31 302616'20 25 1,71 24 10 14 13 13 23 12 23 23 923 8 2.61 45 24 27 38 42 36 15 17 30 34 36 29 26 22 21 24 36 31 45 28 34 28 29 34 15 25 35 32 32 33 22 22 2O 41 24 47 238 60 1 32 18 26 31 19 23 19 31 33 32 30 22 23 20 27 23 34 41 29 40 21 231 32 33 32 29 16 21 13 37 39 34 26 20 32 19 33 35 34 46 24 31 37 33 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1018 25 32 21 20 8 39 34 1 F test only run where SVilks passed F Test Test B vs Control F Test Test C vs Control F Test Test D vs Control F Test ' F Test Test E vs 1 Test F vs Control Control F Test Test G vs I Control F Test Test A vs Control F Test Test I vs Control 1 1 2.77 3.72 4.36 26.57 2.04 6,76 1.11 1.50 3.92 2,56 8,24 15,51 23,36 4,55 30,6.50 4 * 4,74 4,16 1.05 I.63 12,30 2,98 1,34 14.86 3.86 8.14 12.30 11,10 9.61 3.48 12.04 4.93 27.21 2,13 1,85 1.14 11,79 F test only run Where SVilks passed w 1 1 3.68 2.74 2.34 2.60 5.01 * I,51 11.31 6,79 2,60 3.98 1,24 1,52 2,29 2.24 2.97 1,51 2,15 2,45 10,72 w 1.21 3,43 7,62 1.46 2,65 1.11 1,21 1,09 1.06 2,93 1.18 2,01 2.67 3,74 1,32 1,16 F test only run where SVilks passed 1.69 1.26 1,08 5.67 * 2,30 1,44 5,20 3.12 1,20 1.16 3,31 4,98 1.03 6,47 1,39 1.01 1,13 4.93 2.62 1.57 3.50 3.17 1.22 1,86 2,62 2.37 2.05 1,35 2.51 1.05 5.80 1,12 8,69 1.65 2.51 F test only run where SVilks passed 1 1 1.55 2,09 ,, 2,44 14.89 * 3.19 1,98 1.I9 2,20 1.43 4,62 8,69 13.09 2.55 31640 2.66 2,33 1,88 609 .89 1.67 1.33 8,32 2,16 4.90 6,89 6.22 5.38 1.95 6.15 2,16 15.24 1,53 3,31 4.34 6.61 F test only run where SVilks passed 1 1 w 1 4.90 F test only run where SVilks passed 1.58 1.17 1.00 6.09 * 2.14 1,55 4.84 2.91 1.11 1,71 1.89 3.55 5,35 1,04 6,95 1.49 1.09 1,05 4.59 2.82 1.47 3.26 3.40 1.13 2,00 2,82 2.54 2.20 1.25 2.76 1.13 6.23 I.60 8,09 I,11 2,70 F test only run where SVilks passed 1 .1 1 3.65 2,72 2,32 2,63 4.97 1.50 11.22 6,14 2,58 3.95 1.23 1.53 2.31 2.22 3.00 1,56 2.13 10,64 1.22 3.40 7,56 1.41 2.62 1.16 1.22 1.10 1.05 2.90 1.19 2.05 2.69 3,71 1,3I 1.11 F test only run Where SVilks passed 1 1.24 1 1,08 1.27 7.74 * 1.69 1,91 3,81 2,29 1.14 2,40 4,52 6,81 1.33 8,84 1,90 1.38 1.21 3.61 3.58 1.15 2,56 4,33 1.12 2.55 3.58 3.23 2.80 1.01 3,51 1,44 7.93 1,26 6,36 2,25 3,44 F test only run where SVilks passed 1 1 Appendix C F Test of all controls vs test groups failing the mini -chronic test on reproduction * = Non Homogeneous Variance •ple L Test .r D. t 1 Date -21 -28 -29 -30 -32 Y-1 Y-13 V-15 Y-I 1 V-4 V-5 V-6 V-1 '-12 'I-4 'I-S /I-6 'I-7 II-8 II-9 II-10 II-11 /I-I2 III-1 III-2 III-3 I-15 I-21 07/19/88 06/01188 0 j06j89 OS�OB189 OSIZ9�89 06J26j8 OB109188 08/23/88 10123I89 12/06/89 12/13/89 09Ia4j8 08116►89 09fa1189 10/09/89 11 29 89 Ol 10 89 1016/89 I1j01%89 11/02/89 01110190 01/17/90 1 30 27 25 14 2$ 16 20 16 26 35 26 23 24 16 24 13 15 30 23 21 Control Group Reproduction 1 Test : Test A vs 2 16 25 39 12 33 24 16 3� 16 22 33 34 22 13 0 17 24 22 15 3 23 26 23 30 20 21 11 22 42 32 19 118 25 23 16 21 14 13 20 4' 5' 6' 30:22:29 33119120 35142' 34 18120 3 26:43 34 241 9 18 13119 14 15116 15 29:28 34 20:30 29 I5115 17 11116 15 25:28 35 20111 18 32123 22 33134 33 29124 24 11124 29 20124 22 21123 21 21:21 20 141 8 16 29 25 24 32 29 22 14 17 15 19 21 15 19 15 21 15 20 11 23 25 23 1 36 25 37 15 30 16 17 23 16 18 31 21 28 1 26 24 22 19 17 20 49 18 32 17 23 8: 9 30133 25:31 30136 19112 31130 13110 14:21 16123 38126 32:31 16:11 11115 24139 22120 32127 30' 30 28 26 20 23 21 18 19 28 32 24 20 15 23 21 0 21 15 18 17 17 16 25 21 15 24 24 101111121 Control 2431=301 28 29131 38 43140 19 201 81 30 3I1291 23 4211 15 16 151 23 24 29 36 33 24 35 28 31 14 15 18 15 16 16 29 34 33 18 18 15 18 26 23 38 29 31 12 19 31 25 27 24 21 23 22 22 26 29 26 18 24 15 23 14 14 10 23 36 33 20 20 16 15 11 16 15 4 30 27 13 16 16 19 22 20 Test Group Reproductio A. 8 D - 1 G 8 24 23 28 14 6 12 26 41 20 29 30 18 6 20 30 42 11 21:30 11132 36129 14112 16120 19:13 26131 38:40 1 UI i 1 1 33 26 26 23 4 36 38 29:25127 141121 0 33133121 9:22116 18124125 101111 6 31:21132 38:36:30 19111:16 21:25 11116 28132 12:21 14 5 18 34 29 36,38 10110 1 1 1 1 1 1 1 1 28 20 26 12 19 6 41 38 25 0 32 21 20 8 39 34 8,7 1,91 1.15 1.63 2.15 1.80 4.46 2.35 .870 0,09 1.66 * 80 .36 .96 3.08 2.32 1.84 1,31 1,26 2.85 4.17 1.99 4.36 1,35 2.01 1 test only run where SVilks passed 1 Test Test B vs Control 3,40 5,80 4.12 3,12 3.72 1.50 15.25 * 4.98 3.58 3,12 1204: 4,93 1,03 3.41 2.11 15.51 3.64 5.10 5.32 2.35 1.85 21.93 13,34 1.53 9.02 13.46 1 Test Test C vs Control 3,00 1.16 2,48 3.21 2,14 6.19 1.49 2.05 2,85 2.14 6,62 11.59 2.93 1.18 2,01 2.99 1.52 2,81 2.00 1.92 4.34 2.74 1,31 6,65 1.13 1,32 w 1 Test Test D vs Control 1,38 1.24 1.14 1,50 1,26 3.12 3,25 1.06 1.31 1.26 14.40 25.21 1.35 2,51 1.05 1,37 2.16 3.31 1.29 1,09 1.13 2,00 8.69 5.96 2.84 3.06 1.92 2.87 1 Test 1 1 Test ' 1 Test Test E vs 1 Test 1 vs Test G vs Control 1 Control Control 1.91 1 3.25 2,31 1►75 2.09 1.19 8.55 2.19 2.01 2►09 31.85 66.23- 1.95 6.15 2.16 1.91 1.22 8,69 2.04 2.86 2.98 1.32 3,31 15,65 1.47 1.16 5,05 1.54 1.28 1.33 1.06 1.40 1.11 2.91 3.49 1.14 1.22 1.17 15.47 27.01 1.25 2.16 1.13 1.28 2.01 3.55 1.20 1.11 1.22 1,86 8,09 6,40 3,05 2.85 2.01 3.08 1 Test : 1 Test 1 Test 8 vs 1 Test I vs 1 Control 1 Control 2.97 1 2.91 1.74 1.74 2.46 2,46 3.24 3.24 6.14 * 6,14 1.51 1.51 2.03 2.82 2.82 2.12 2.72 6,61 * 6.61 11.68 2.90 2.90 1.19 1,19 2,05 2.05 2.91 1.53 2.18 1.99 1.90 4.31 2.16 1.32 6.60 1.12 1.33 2.97 4.66 1,53 2.18 1.99 1.90 4,31 18.75 2.16 1.32 6.60 1.12 1,33 1 test 1 test 1 test 1 test 1 test 1 test 1 test 1 test only run only run only run only run only run only run only run only run where where where where where where where where SVilks SVilks SVilks SVilks SVilks SVilks SVilks SVilks passed passed passed passed passed passed passed passed a aaple "1 Test .D. t : Date I-1 I-2 I-3 I-4 I-6 I-7 I-8 I-9 I-10 I-11 1-13 I-14 I-16 I-17 1-18 I-19 1-20 I-21 I-22 1-23 I-24 II-1 III-1 IV-1 IV-2 IV-3 1V-4 IV-5 IV-6 V-1 Y-2 V-3 YI-1 VI-2 VI-3 I-25 I-26 06J14188 O1115186 1010 189 1012SI86 03/13/89 � 05/08/89 OS j09189 0T U 89 08 OT 89 0/02/89 0/17/89 0 (189 0/18/89 II/(306 1 89 21 2118 212TI89 2/27/89 0/17/88 0/23/89 1�/5�88 09/11/89 10/12/89 10/16/89 IO2)I889 12�06 /9 12 1J 89 06/08/88 03/14/89 10/16/89 01/08/90 01/08/90 WIZ1= Test A vs 1 1 45 24 11 27 38 28 24 29 45 2 31 41 51 61 1i 81 9110111112i- Control, , 21134 32'34 28127 22 15 36 29 290.9672 41136 42 34 46140 31 31 29 42 39 0.9738 11116 19 15 19110 17 11 11 11 19 0.9338 291302926281292932292928 0.9546 19130 I1 34 20120 36 34 18 31 28 0.9496 811620161216121021 1815 0.9774 10:13 35 19 34 12 37 20 36 31 39 0.9659 32132 31 35 31 26 29 24 23 2I 0.4332 24130 24 9 18 25 13 10 23 4 21 0.9822 22121 34 24 31 31 30 26 16 20 25 0.9797 10114 13 13 23 12 23 23 9 23 8 0.9205 3124 4541 24128 24121 21134 38133 335128 2 42129 31 36 19 15 17 30 36 29 26 22 21 24 41 29 34 13 15 25 35 32 32 33 24 22 26 22 2� 37 41 28 30 32 11 26 31 19 23 22 33 30 322 23 20 27 23 34 41 29 40 28 31 33 34 46 30 24 31 31 33 1932 �d0 29 27 11 31 16 10 27 3 21 22 Il 22 )1 79 39 73 ]4 41 26 32 25 28 20 17 32 23 19 IS 13 16 32 29 26 28 JS 43 27 43 33 41 27 31 33 26 31 32 33 31 37 22 32 33 34 38 30 36 33 37 11 18 2723 20 18 31 28 31 40 36,33 32138 30:31 35131 18121 22128 22:20 17118 22149 38:29 43139 31 40 25 21 33 41 29 11 5 30 14 23 17 229 334 33 24 22 19 325 35 31 44 32 46 29 30 29 31 30 32 29 37 26125 28132 4l, 38 27127 30135 24' 25 2244 37 36 28 39 20 20 23 23 30 33 35 329 18 18 21 27' 36 36 36 40 44 29 41 26 32 329 21 18 34 34 20 23 8 36 33 34 18 21 28 33 37 37 22 42 21 33 36 39 35 31 2 30 18 18 33 36 38 15 19 20 36 0,9703 0,9732 0,9127 0.9469 0,9645 0.9605 0,9725 0.9395 0,9064 0.7975 0.9118 0.9587 0,9031 0.9754 0.9164 0,962I 0,9621 0.9121 0.9712 0,9501 0.9108 0.9570 0.9520 0,9735 0.8757 0,9280 0.9712 w 1 Control Group Reproduction 1 SRilks 1 SVilks Test 13 vs 1 Control 0.9671 0.9731 0.9511 0.8987 0.9682 0.9564 0.9309 0.6571 * 0,9651 0.9114 0,9563 0.9476 0.9216 0.9130 0.9434 0.9033 0.9541 0,9678 0.9510 0,9296 0.9203 0.9352 0.9534 0,9225 0.9269 0.9654 0.9453 0.9317 0,9360 0.9434 0,9698 0.9324 0.9631 0.9022 0,9144 0.9540 0,9397 0.9366 SVilks Test C vs Control .9101 .9162 .9384 .9689 .9660 .9718 .9666 .4037 .9857 .9831 .9254 .9774 .9367 .9543 .9217 .9659 .9329 .9703 .9I51 .8941 .7606 .9699 .9398 .8924 .9480 .9640 .9353 .9673 .9283 .9596 .9522 .9498 .9587 .9701 .9821 .8558 .9309 .9761 1 1 Rifts . Test D vs Control 0,9608 0,9588 0.9573 0.9641 0.9290 0,9489 0.9603 0.4612 0.9626 0.9612 0,8114 0,9615 0.9668 0.9733 0,9504 0,9716 0.9248 0.9421 0,9483 0,9026 0.8296 0.9653 0.9164 0,9080 0,9614 0,9513 0.9379 0.9568 0,9470 0,9541 0.9161 0.9591 0.9404 0,9133 0,9555 0,8932 0.9424 0.9706 1 i 1 SVilks Test £ vs Control .9388 .9323 .9546 .9300 .8812 .95 9173 .5762 .9180 .9362 .9113 ►9395 .952t. .9445 .9355 .9360 .9569 .9436 .8996 .8879 .8960 .9548 .9443 .8999 .9565 .9519 .9609 .9508 .9601 .9602 .9357 .9600 .9540 .9307 .9303 .9431 .9236 .9550 SVilks 1 SVilks 1 SVilks 1 Test F vs 1 Test G vs 1 Test R vs Control ; Control Control .8352 .8139 .7791 .T130 .8262 .1851 .8852 .4127 .8806 .8142 * .7866 .7831 .7361 .1261 .7994 .1164 w .7763 w .8043 .8026 ,8513 .7788 .7541 w .8263 .8500 :7354 .8038 .7106 .7551 .1526 .7108 .8132 .1382 .8029 .1047 .8217 ,8761 .7659 .7609 w1 *1 w: w: w Test Group Reproductio SVilks Control vs Control 1 F Test Eq T Test Vilcoxon A` 8 F G 1 1 24120 23129 28130 14118 61 6 12120 26:30 41142 8:11 21130' 33 29125127 11132 26 141121 0 36129 26 33133121 14112 23 9122116 16120 4 18124125 19113 101111 6 26131 36.31:21:32 38140 38138:36130 61 6 21:19:17:16 1 1 , , 1 1 1 , , 1 , , 1 , 21:25 11116 28132 12121 141 5 71 8 34129 36138 10110 28125 201 0 26132 12121 19120 6, 41139 38134 81 7 I-10 vs I-11 1-17 vs I-18 I-11 vs 1-19 I-18 vs I-19 I-23 vs 1-24 I-25 vs I-26 0.9283 0.8515 w 0,9413 0,9130 0.7468 0,8943 1,53 1.43 4.66 1,52 -4.19 ; 4.82 3,15 -4,11 98.5 116.0 • 0.9112 0.9814 0.9218 0,9372 0.9404 0.9144 0.9593 0,4801 0.9801 0.9818 0.9064 0.9759 0.9634 0,9547 0,9705 0.9448 0.9525 0,9147 0.9756 0.9169 0.8408 0.9602 0,9687 0,9241 0.9575 0.9577 0.9579 0.9506 0.9575 0.9603 0.9400 0,9525 0.9483 0.9361 0.9558 0.8959 0.9427 0.9632 .9486 .9569 .9416 .8980 .9451 .9481 .9560 .3951 .9722 .9643 .9411 .9190 .9539 .9468 .8849 .9328 .9699 .9666 .8614 .8892 .7501 .9518 .9I69 .8882 .9640 .9717 .9651 .9458 .9600 .9696 .9628 .9750 .9559 .9118 .9817 .8554 .8861 .9467 1 1 1 1 SVilks Test I vs Control 0,9565 0,9566 0.9205 0.9563 0,9055 0.9483 0.9450 0,4967 0,9521 0.9543 0.8630 0.9621 0.9556 0.9662 0,9736 0,9659 0,9143 0,9404 0.9111 0.9048 0.8643 0,9484 0,9732 0.9094 0.9534 0,9318 0.9303 0.9428 0.9378 0.9381 0.8912 0,9493 0.9305 0.9638 0,9259 0,9060 0,9261 0.9532 1 1 1 Appendix D Shapiro -Wilkes Test Results comparing distribution of all control groups vs the test groups failing the mini -chronic procedure on reproduction * = Data are abnormally distributed *pie D1 Test Date -27 -28 -29 -31 1 -32 Y-1 V-13 Y-15 Y-11 v-4 Y-5 Y-6 Y12 I-5 'I-7 I9 "I-I0 'I-11 1-12 `II-1 'II-2 'II-3 '-15 '-21 06101188 02/20/89 03/06/807/10/89 05/08/89 OS j29189 06/19/89 08�0 %88 1008�2388 23 j89 12106/89 11106�89 09/OdI89 OB( 01189 09/11/89 10 09 89 U�29 89 01/10/89 1t101189 11/02/89 11/30/89 01/10/90 O1I11190 1 30 27 14 22 I8 16 20 I6 16 26 35 27 26 23 9 22 16 24 13 15 30 23 21 Control Group Reproduction : 2 3: 4 5 6: 7 8: 9 10'11:121 16 23' 30 22 29:36 30:33 24 31:30 25 26 33 19 20:25 25:31 28 29:31 39 35 35 42 34:37 30:36 38 43:40 1223 1820 3:15 19:12 19 20: 8 33 33 26 43 34 30 31:30 30 31129 24 30 24 9 I8 25 13:10 23 4'21 13 19 13 19 14 16 14:21 15 16 15 16 20 15 16 15 17 16123 23 24 29 3021,2928342338:26 36 33 24 31 21120 30 29:34 32:31 35 28 31 16 18115 15 17:I6 16:17 14 15 18 16, 11117 16 15:18 17115,15 16 16 24:37:25 28 35:31 24:39:29 34, 33 22:22:20 11 18:21 22:20:18 18:15 26123132 23 22:28 32:27:18 26:23 33:42:33 34 33: 1 30:30:38 29:31 32:29:29 24 24:26 28:26:12 19:31 5' 32:17 24 29:26 20:23:25 27:24 19 19:20 24 22:24 21:18:27 23:22 22 19' 21 23 21:22 19:28:22 26:29 27 31 21 21 20:19 32:24:26 18' 24 13 18 14 8 16:17 20:15 15 23 14 1 25 29 25 24:20 23:21 14 10 23 0 23 32 29 22:49 0:27 36 33 20 16 16 14 11 15:18 15:18 20 16 15 17 21 19 21 15:18 17:17 11 16 15 24 14 19 15 21:32 16'25 4 30 27 22 13 15 20 17:11 21 15 13 16 16 1520232523232424192220 Slinks Test A vs Control 0.9578 0.9102 0.9706 0.9643 0.9746 0,9822 0,9753 0.9315 0.9126 0.9610 0.955I 0.9349 0.9501 0.9108 0.9570 0.1211 0.9415 0,8981 0.9814 0.9454 0.9520 0.9742 0.9659 0.8757 0.9735 0,9673 0.9591 0.9562 0,9735 w SVilks Test B vs Control 0.9621 0,9608 0.9639 0.9674 0,9596 0.9651 0,9253 0,9420 0.9733 0.9682 0,8837 0,8106 0.9698 0,9324 0.9631 0,8895 0.9523 0,9388 0.9265 0.9498 0.9604 0.9573 0.9611 0.9540 0.9066 0.9292 0.9554 0.9411 0,9296 SVilks Test C vs Control 0.9522 0,9761 0.9623 0.9567 0,9638 0.9857 0,9369 0.8919 0.9853 0,9568 0,9653 0.9622 0,9522 0,9498 0,9581 0,6915 0.9205 0,8725 0,9512 0,9215 0,9594 0.9686 0,9615 0.8558 0,9645 0.9812 0,9566 0,9318 0,9768 1 1 Test Group Reproductio SVilks Control vs Control ti 24 23 28 14 6 12 41 20 29 30 18 20 42 21:30 17:32 36:29 14:I2 16:20 19:13 26:31 38:40 33:29:25:21:21:25 26:14:12: 0:11:16 26' 33:33:27' 28:32 23: 9:22:16 12:21 418:24:25 14: 5 10:11: 6, 7: 8 36 31:27:32:34:29 38 38:36:30:36:38 28 20 26 12 16 41 38 : 8:17: 6: 6:21:19:17:16:10:10: 8 , 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 , 1 , 1 20 32 21 28 34 7' I-10 vs I-11 I-13 vs 1-15 I-17 vs 1-18 1-23 vs I-24 I-25 vs I-26 0.9283 0.9400 0,8515 0.7468 0.8943 SVilks 1 SVilks Test D vs 1 Test E vs Control Control 0.9590 0.9501 0.9608 0,9492 0.9799 0,9626 0,9558 0,9017 0,9553 0,9551 0.9650 0.9533 0.9161 0,9591 0.9404 0.7690 0.9440 0.9175 0,9632 0.9352 0.9421 0.9551 0.9515 0.8932 0.9113 0.9688 0,9562 0.9312 0.9761 1 0.9273 0.9435 0.9362 0.9300 0,9530 0,9180 0,9549 0.9516 0,9312 0,9158 0,9210 0,9065 0,9357 0,9600 0.9540 0.8416 0.93I3 0.9279 0.9553 0.9433 0,9633 0,9540 0.9096 0,9431 0.9417 0.9481 0,9414 0.9595 0.9419 i SVilks 1 Test.F vs Control SVilks : SVilks 1 Test G vs Test H vs : Control Control 0.8082 *: 0,8022 w, 0.8222 *: 0.8288 0.8181 0,8833 =: 0,7366 *1 0,1637 0,8176 w, 0.8234 * 1 0.6790 * 0.6622 *1 0,8166 *1 0,7370 w) 0,8028 0.7454 *1 0,8203 *1 0,8185 *1 0.7341 *1 0.8199 *1 0,7900 *1 .8460w1 0 0.8101 * 0.7162 *: 0,1534 ^: 0.8592 *: 0,7570 *: 0.7504 *: 0.9115 0,9713 0.9826 0,9189 0,9666 0,9801 0.9541 0.9103 0.9642 0.9762 0,9224 0,9036 0.9400 0,9525 0,9483 0,7725 0,9721 0,9260 0.9586 0.9650 0.9407 0,9631 0.9740 0,8959 0.9452 0,9490 0.9560 0.9486 0.9679 1 0.9405 0.9383 0.9392 0.9397 0,9522 0.9722 0,9658 0,9556 0,9709 0,9231 0,9145 0,8807 0.9628 0,9750 0,9559 0.6794 0,8868 0,8506 0.9673 0.9060 0,9805 0,9629 0.9401 0.8554 0.9477 0.9387 0,9502 0.9723 0,9221 1 1 1 1 SVilks Test I vs Control 0,9519 0,9514 0.9696 0.9507 0,9688 0.9527 0.9459 0.8805 0.9339 0.9648 0,9600 0.9510 0,8912 0.9493 0,9305 0,8029 0.9638 0,9285 0.9562 0.9575 0.9161 0,9509 0.9501 0.9060 0,9622 0,9518 0.9408 0.9228 0,9686 Sample I.D. I.D. 1 I-13 I-14 I-16 I-17 I-18 I-19 I-20 1-21 I-22 I-23 I-24 II-1 III-1 IV-1 IV-2 1Y-3 IV-4 IV-5 IV-6 V-1 V-2 V-3 VI -I VI-2 VI-3 1-25 I-26 test Date 06/14/88 07/15/88 08/21/88 10/03/89 10/17/88 10/25/88 03/13/89 04/24/89 05/08/89 05/09/89 05/09/89 07/11/89 08/01/89 0/02/89 0/17/89 0/11/89 0/18/89 1/06/89 1 27 89 2/13/89 2/27/89 2/27/89 0/17/88 0/23/89 0/03/88 1/15/88 09/11/89 0/12/89 0/16/89 0/23/89 0/23/89 2/06/89 2/13/89 06/08/88 03/14/89 10/16/89 01/08/90 01/08/90 Control Group Reproduction I 26 45 27 24 31 11 27 31 8 28 24 1014 29 3124 45 45 47 24 28' 28 24 27'30 27 34' 30 38 33'41 35 28' 32 29 30 42 29' 11 18 31 29' 26 36:34'31 19'13'19 I5115'23 27124'22 17:25: 19 30' 35 31 3432 33 36, 32 32 29' 33 30 27124 31 26, 22 22 18' 26' 23 22122'20 21124'27 24 0' 23 36:37' 34 4I:4I'41 2' 3: 4: 5: 6: 7: 8' 9' 10' 11' 12' I 1 1 1 1 1 1 1 21'34'32:34 411364234 11I 29' 30 29 26 19 30 17 34 8' 16 20 16 1 13 35 19 32' 32 3l : 35 24' 30 241 9 22' 27 34; 24 13:13 29:34 40; 46 28130 21:24 31131 32:37 33133 32 : 32 19140 29:27 27:31 16 10 27 3 21 22 13 22 31 39 39 33 34 41 26 32 25 28 20 17 32 23 19 15 13 16 32 29 28 28 35 43 46 19 28 20 12 34 1 18:25 3l:31 23112 21 43 21 33,26 31 : 32 33:31 31122 32:33 34 38 30 33 11 27 20 31 36 32 30 35 18 22 34133 36 : 36 33:38 33,28135 35129 24139129 34133 22120118 18'1S 32:27;18 26 23 22.20 19:20:21,21 19 17:18 25:31:32128 19 22:49 0:27:36 33 20 38129 35:36:36 37 36 43139, 38:40144 37 45 27' 22115136 29:29 40 37131129 42:39 (0 11111:17 11119 29 29:32:29 29128 203613411831128 16 12110121 18115 1237120:3631139 26129, 24 23121 13110:23 4:21 30:26 16 20125 23: 23 9 23: 8 33 31:31 32 29:22 41 40144 46 41:42 3125293030:27 27293126114 33 30 32 30 : 33 41293732138 29262529136 35128132 21:30 36 41; 38 18139 5 21127 34' 35 30301353431 14 26:16 20 2 23 4125 23 30 202012324,18 11 24123 18 18 3537:30 29 39133 34 36:39 36 31 18 23 18 28 40 33 38 37 37 28 1 I 1 1 1 1 1 1 1 1 1 test Group Reproduction A '6 C •' D F C 8 24' 20:2I 30133 29 25:27 23 29:17 32:26 14 121 0 28 30:36 29126 33 33127 14 18:14 12 23 9 22:16 6 611620 4 18 24:25 12 20:19 13 10 111 6 26 30126 31 36 31 27132 41I42138 40 38 38 36130 8:17: 6: 6 21119:11;16 1 1 1 1 1 1 1 1 1 21 11 28 12 14 7 34 36 10 25:28:25 16:201 0 32126132 21'12:21 5 19120 8 6: 8 29 41139 38, 38:34 101 8: 7 1 1 , 1 1 I Eq T test Test A vs Control 0.99 6.75 -5,34 * 2,23 0.35 -6.60 ^ -3.11 0,21 -4,36 2.40 13.35 1.81 0.41 4.38 5.86 2.69 2.00 4.78 -4.80 -2.98 -2.35 5,74 6.19 6.17 3,88 2,74 -4,05 -0,48 -4.46 -1.46 5,47 10,48 Eq T Test only run where F test passed Eq T test Test B vs Control 3,23 6,59 -0 , 20 2.74 ^ 2.20 0.20 2.88 w "-0,12 3.03 ^ 4,25 * 3,87 3.66 3,03 -0.25 0.81 1.39 4.29 * 1,12 1.59 Eq T Test only run where F test passed Eq T test Test C vs Control -1.13 4.71 -8.32 * -1.38 -(0.23 ^ -2,19 -6.69 * -0.48 11.66 * -1.14 -2.11 1.10 3,50 0,25 -0,39 1.94 -7.31 ^ -7.04 ^ -4,98 3.20 3.54 4.25 ^ 1,07 0,55 -8.05 = -3.20 -9,26 ^ -3.70 ^ 2,97 8,41 Eq T Test only run where F test passed EgTtest 1 Eq T test Test D vs : Test E vs Control 1 Control 5.20 *1 10.93 -0.09 4,19 ^ -0.61 0.45 4,91 * 7.74 ^ 17.84 * 5.34 * 10.66 w 7.44 6.64 ^ 10.18 ^ -0.15 1.27 3.48 * 2,51 ^ 10.79 ^ 11.43 * 11,88 9.20 w 7.13 ^ 2,34 4,57 * 2.86 ^ 10.45 * 15.20 * Eq T Test only run where F test passed 4,60 8.81 0.48 0.13 2.98 0,82 4,30 0,52 6.00 4.52 8.21 6.06 5,54 4.64 4,16 0,39 1.51 2,44 8.10 6.00 3,92 2.77 2,15 7.95 1 Eq T test 1 Eq T test Test F vs : Test G vs Control ; Control 4.30 Eq T Test Eq T Test only run only run where where F test F test passed passed -1.81 3.21 -7.95 -2.14 -9.27 -1.69 -5.18 -2,71 - 6,73 -1.44 7.53 -2.71 1.83 - 0.13 -1,24 0.08 0.36 -7.24 -4.13 - 6,40 - 5,15 1.45 1.58 2.16 -0,25 -0,39 -7.25 -3,62 -4.07 1.33 5.31 Eq T Test only run where F test passed Eq T test Test R vs Control -4.85 0.61 -12.11 -8.20 -15.28 -6,43 -10.28 -5,86 5.40 -8.43 -6,51 -5.85 -1,52 -4.28 -11.13 -13.20 -9.18 -2.27 -2.40 -1.56 -4,53 -3.51 -13.95 -1,19 -16.08 -7.39 -2.31 2.51 Eq T Test only run where F test passed Eq T test 1Test Ivs1 i Control 6.59 Al 12.00 1.84 11 1.85 11 *1 1 11 11 ,I 11 ^1 1 ^1 1 1 w, ^1 1 1 I ^ ^ 5.54 1.54 4.02 6,47 9.13 18.09 6.85 11,76 8.19 8,04 5.87 11.32 1.61 2,58 5,39 4.20 11,87 12.43 12.84 10.45 8,45 4,35 6.17 4.40 11,57 15.79 Eq T Test only run where F test passed Appendix E Equal variance T-Test results, unequal variance T-Test results and results of rank sum tests. * = Test Group and Control Groups are statistical different. Savple I.D. I Test Date Control Group Reproduction 1 2 1-27 I-28 I-29 1-30 I-31 1-32 IV-7 IY-13 IY-15 1V-11 v-4 Y-5 v-6 Y-1 V-12 VI-4 VI-5 YI-6 VI-1 VI-8 VI-9 VI-10 VI-11 VI-12 YII-1 YII-2 VII-3 v-15 v-21 01/19188 06/01/88 02/20/89 03/06/89 01110189 05/08/89 04/24/89 05/29/89 06119189 06/26/89 08/09188 08/23/88 10123/89 12/06/89 12/13/89 11/08/89 09/04/89 08/14/89 08/07/89 09/11/89 100989 11 29/89 07/10/89 10/16/89 11/01/89 11/02/89 11/30j89 01 / 10/90 01111190 3:4:5 316 23 322 270 25 26 33 19 25 39 35 35 42 1412231820 22 33 33 26 43 8 24 30 24 9 186 16 20 15 16 33 30 21 29 28 20 31 27 20 30 16 16 11 11 16 21 24 31 25 28 26 22 22 20 17 1826233223 35 33:42, 33 34 21 32:29:29 24 26 5132111 24 23 19:19:20 24 9 22:19:21 23 22 27:31:21 21 24:13:18:14: 8 16: 7:25:29:25 24: 0 23:32 29 13:16 16 14 17 15:11 21 19 21 30:24 14 19 15 23:22 13 15 20 21115202325 61 1' 81 9;10;11;12 29:36 20:25 34131 3:15 34130 18:25 14:16 15,11 34 23 29 34 11 16 15 18 3537 18 21 22 28 33 1 24 26 29 26 22 24 27:22 20:19 16:17 24' 20 22 49 15 18 15,18 21:32 I1:17 23:23 30: 25 30 19 37 13 16 38 32 16 17 24 22 32 30 28 20 19 32 20 20 15 16 21 24 33 24 31 28 36 38 12 19 30 30 10 23 21 15 23 23 26 36 31 35 11 14 15 15 39 29 20 18 21 18 30 38 26 12 23 25 18 21 28 22 24 26 15:15 21:14 27:36 18 11 25 15 24 20 11 4 13 19 Eq T test : Eq T test ' Eq T test Eq T test 1 Eq T test : Eq T test : Eq T test Test A vs : Test B vs Test C vs 1 Test D vs 1 Test E vs 1 Test F vs : Test G vs Control ; Control Control Control S Control S Control ; Control EgTtest :EgTtest 1 Test H vs 1 Test I vs Control Control 31:30 29 31 43 40 20 8 3l4 16 24 33 15 16 34 18 26 29 19 21 26 23 33 16 16 16 22 29 21 15 29 24 37 18 16 33 15 23 31 31 24 22 29 14 20 15 15 16 20 Test Group Reproduction A 8 C E F G H 24:20 23129 28:30 14118 6: 6 12120 26' 30 41:42 8:17 21130 33 17:32 26 36:29 26 14:12 23 16:20 4 19:13 26:31 36 38:40 38 6: 6 21 1 I 29 14 9 18 10 31 38 19 25:27:21:25:28:25 12: 0:11:16:20: 0 33:21:28' 32:26:32 22:16:12'21:12:21 24:25 14 5 19:20 11:6186:8 27:32 34 29 41:39 36:30 36 38 38:34 17:16 10 10 81 1 1 1.11 0,55 5.70 -5.16 * 3.01 -3,11 -1.03 -3,73 2.03 2.02 2.14 -4.05 = -0.48 -0,04 -1.09 -2,82 - -1,78 -0,82 -5,41 -2.65 -1.51 - -6,34 -1,61 -5.55 -2.93 Eq T Test only run where F test passed 3,34* 5.94 -0,42 4.44 0.20 3.88 3.81 4.29* 3.41 2.61 1.83 1.68 0.88 1.59 1.26 -1.18 -2.23 3,59 -7,15 0.84 -11.69 -6.73 -0.23 -0.28 5,57 : 4.81 5.10 * 4.10 10.06 : 8.12 -0.43 0,18 1.51 = 6.25 = 0.45 0.82 -0.05 1.58 1.72 6.50 * 5.50 6.53 * 5.51 = 0.55 1.13 * 6.00 = -8,05 2,34 -3.20 * 4,51= 3.92= -2.40 4.50 * 3,99 = 2,90 * 2,83 -6,89 3,65 -4,31 2.96 * 2.79 -3.60 4.29 * 3,11 = -8,63 0,13 0.65 -4.81 : 1.52 1.111 2,15 -12,93 -10,76 • 0,42 1,92 2.06 -9,42 * 0.70 -6.89 : 3.43 * •. -1.88 -5.20 * 1 1.00 * -2.82 * -7.01 7.24 2.14 -0.73 11.20 = -7,60:-11.63:; 1,38 -0.16 -3.32 8.82 -5.18 = -9.86 * -11,88 2.28 -6.57 * -11.33 -1.01 -4.33 = 1.88 -1.12 -4.44 1.91 I i -0.39 -3.51 8.45 -1.25 =: -13.95 4.35 -3.62 *-7.79 :; 6.11 -2.94*-6.42:; 6.00= -3.55 4.35 -6,21 -13.11 5.56 -4.59 :1 -8.43 *' 4.59 -3,95 -8.24 5.92 -8,11 -13.33 2,13 -5,08 :1 -8.29 *1 3.08 *1 -19.16 * : 2.38 -9.25*:-16,18=: 2.67:' -3,17 *: - 8.45 *' -14.91 :: 2.81 *� - 6,33 -12.93 5.34 Eq T Test Eq T Test Eq T Test Eq T Test Eq T Test Eq T Test Eq T Test Eq T Test only run only run only run only run only run only run only run only run Where where Where where where , where where Where F test F test F test F test F test F test F test F test passed passed passed passed passed passed passed passed Control Group Reproduction 2 3 4; 5 6 1 8 ; Unequal Test ; Unequal Test Test A vs Control Test B vs Control 9:10:11:12: Value df 1 Value df Unequal Test ; Unequal Test ; Unequal Test Test C vs Control ;Test D vs Control ;test E vs Control Value df ; Value df Valve df Unequal Test Test F vs Control Value df Unequal Test Test I vs Contro Value df Sasple I.O. 1 Test Date 1 I I Unequal Test ; Unequal Test Test G vs Control test H vs Control Value df 1 Value df 1-1 I-2 1-3 I-4 1-5 1-6 I -8 1-9 1-10 I-I1 -13 -14 1-16 I-11 1-18 1-19 1-20 I-21 1-22 I-23 I-24 II-1 11I-1 IV-1 1V-2 IV-3 IV-4 IV-5 IY-6 V-2 V-3 VI-1 VI-2 VI-3 1-25 1-26 06/14/88 01/15/88 08/21/88 10/03/89 10/17/88 10/25/88 03/13/89 04/24/89 05/08/89 05/09/89 05/09/89 01/11/89 08/01/89 0/02/89 0/17/89 0/17/89 0/18/89 1/06/89 2/13/89 2/27/89 2/27/89 0/11/88 0/23/89 0/03/88 1/15/88 49/ 11 /89 0/12/89 0/16/89 0/23/89 0/23/89 2/06/89 2/13/89 06/08/88 03/14/89 10/16/89 01/08/90 01/08/90 26 45 24 31 11 21 41 11 29 18 27,10 31;32 8;24 28122 24:10 29:31 24 29 45145 41 40 24:28 28 28 24:21 30 27 21 34 30 31 38 33 41.32 35 28 32:33 29 30 18:32 42 29,11:19 ,31 29:26:29 :36 34:31:21 119 13:19;16 ;151523:27 :21 24 22:21 117 25 19:13 :30 35 31:31 :34 32 33; 39 :36 32 32:34 293330:26 21 24 31;25 26 22 22;20 18 26 23:32 222220119 ;21 24 27:13 :24 0 23132 ;36 31 34:28 ;41 41 41135 34 36 16 30 16 13 32 30 27 14 32 42 19 29 17 20 35 31 24 34 13 34 34 26 16 19 35 24 13 34 46 28 46 28 20 12 34 31 3l 123 27 43 30.27 24;33 31;31 31;33 33;31 32; 32 40; 34 27'30 31 13 22 22 39 33 41 32 28 11 23 15 16 29 28 43 33 27 31 236 32 30 35 18 22 22 17 22 38 43 21 40 10 16 212 25 31 12 22 37 17 12 326 13 30 23 15 31 11 32 34 10 20 29 26 23 36129 29:42 11;11 29; 29 18:31 21:18 36' 31 24 23 1 6 20 9 23 33 31 31:32 41 40 44; 46 31 25 29 1 30 26 21 29'31 32 33 30 32 31 41 29 31 22 29 26 25 33 35 28 32 38:36:41 38 36: 5:21 21 31:30:30 35 18;14126 16 23:23: 4 25 18:20; 20 23 28 11:24 23 40 35; 31 r 30 33 29 39 33 38 34 36 39 37 33 28 35 37 24 39 29 21 22'20 18 2832;2718 20 19 20 21 18 25' 31 32 49 0; 21 36 29 35.36 39 38' 40 r 29 47 29 39 19 28 28 15 21 21 25 8 22 42 3027 2614 30 33 32 38 29 36 21 30 18 39 34 35 34 31 20 2 23 30 24 18 18 18 34 33 36 36 33 38 35 29 34 33 18 15 26 23 21 19 28 19 33 20 36; 37 36 44'.31 45 r , Test Group Reproduclio A C G 1 ;24 :23 28 :14 :6 :12 :46 20; 21; 30 29:11; 32 30; 36; 29 18:14;12 6116120 20:19113 30:26:31 42:38:40 8,11: 6; 6 , , r 33129125 26:14:12 26:33:33 231 9'22 4 18 24 10 11 36 31 21 38 38 36 21 19 17 27 21;25 0 11:16 27 28:32 16 12121 25r14; 5 6;1;8 32 30 16 r r r 28 26 12 19 6 25 0 32 20 8 34:29r41 39 36:3838 34 10;101 8 7 r r , 0.13 13.00 1.99 14.00 -2.09 • 15.00 Unequal T Test only run where F Test Failed 3.95 -0.50 11,00 14.00 4.01 * 13.00 8.95 * 12,00 3.80 * 11,00 4,85 * 11.00 5.85 * 14.00 5,15 * 12,00 I.62 12,00 5.69 * 13.00 5.80 * 12.00 6.10 * 12.00 4,71 * 13.00 1.06 12.00 2.53 15.00 1.11 11.00 S,bO * 13.00 1,80 * 12,00 Unequal T Test only run where F Test Failed 15,00 12,00 14.00 0,66 13.00 -3.16 * 13,00 Unequal T Test only run where F Test Failed 8.31 2.93 14.00 15.00 6.04 12.00 12,51 =; 13,00 , r , 8,00 15,00; 5,84 =; 12,00 10.21 =; 14.00; 7.24 * 12.00 4.81 15.00 2.61 1,99 Unequal T Test only run where F Test Failed 14,00 13.00 1,50 * 14,00 2,93 = 13,00 8,34 : 14.00 8,11 * 14.00 6.95 : 14.00 2.19 14.00 2.31 12,00 10,93 Unequal T Test only run where F Test Failed 14.00 Unequal T Test only run where F Test Failed -2.19 ; 14.00 -2.44 -0.44 -1.80 -1.11 Unequal T Test only run where F Test Failed 14.00 14.00 14.00 13,00 -1.69 -6.50 Unequal T Test only run where F Test Failed 15.00 12.00 14,00 13.00 13.00 9.73 13. 9.41 =: 14 11,39 *; 13 4.10 2.90 Unequal T Test only run where F Test Failed 13 14 Saiple I.D. 1 Test Control Group Reproduction Date ;1:2:3 I-21 1-28 1-29 1-30 I-31 I-32 IV-7 IV-13 IV-15 IV-11 Y-4 V-5 Y-6 V- 7 V-12 VI-4 VI-5 VI-6 VI-7 VI-8 VI-9 VI-l0 VI-1l VI-12 YII-1 YII-2 VII-3 V-15 V-21 01/19/88 30 16 06/01/88 2l 25 02/20/89 25 39 03/06/89 14 12 01110/89 22 33 05/08189 8 24 04/24/89 18 13 05129/89 16 16 06119/89 33 30 06/26/89 20 31 08/09/88 16 16 08/23/88 16 16 101231P 21:24 12/06/89 26:22 12/13/89 :18126 11 /08/89 :35:33 09/04/89 :27:32 08/14/89 :26: 5 08/01/89 :23:19 09/ 11 /89 : 9:22 10/09/89 :22121 11 /29/89 124113 01110/89 1161 1 10/16/89 1241 0 11/01/89 :13:16 11/02/89 :15:17 11/30/89 ' 30124 01/10/90 23:22 01 /11190 21:15120 4 5 6 8 Unequal Test ;Test A vs Control 9:10:11:12: Value df Unequal Test :Test B vs Control Value df Unequal Test Test C vs Control Value df Unequal Test Test 0 vs Control value df Unequal Test 1 Unequal Test Test E vs Control Test F vs Control Value df 1 Value df Unequal Test Test G vs Control Value df Unequal Test Test 1I vs Control Value df Unequal Tes Test I vs Cont! Value c 23 30 26 33 35 35 23 18 33 26 30 24 19 13 20 1 5 21 29 27 20 18 15 17 11 31 25 22 20 23 32 42 33 29 29 3211 19 20 19 21 31 21 18:14 25129 23:32 2116�19 14 19 13 IS 23 22 19 42 20 43 19 16 28 30 15 16 28 11 29 20 34 34 18 15 34 29 17 15 35 18 23122 34133 24124 24:29 24:22 23121 28: 16 25 24 29 22 11 211 S IS 21 20 11 25 23 36 25 37 15 30 25 16 11 23 34 18 121 28 26 26 24 19 30 25 30 19 13 16 38 32 16 11 24 22 32 30 28 20 21 19 32 17120 20:23 49: 0 18:15 18 11 32 16 11 21 23 24 33 31 36 12 30 10 21 23 26 31 11 15 39 20 21 30 26 23 18 1254 21 21 11 25 15 24 24!31;30 28 29 3I 38 43 40 19 20 8 30 31 29 23 4 21 15 16 15 23 24 29 36 33 24 352831 151616 29 34 33 18 18 15 18 26 23 38 29 31 12 19 31 25 21 24 21 23 22 22 26 29 152314 14 10 23 36 33 20 20 16 15 11 16,15 430:27 13 16:16 19 22120 Test Group Reproduction :24:20 21 123129 17 :28:30 36 :14:18 14 :6:616 :12:20:19 :26:30:26 :41:42:38 8111: 6 30:33:29 25 21121 25 28125 32:26:14 12 0111 16 20: 0 29:26:33 33 21128 32 26:32 12:23: 9 22 16:12 21 12121 201 4 18 24 25114, 5 19120 131 10111 6: 1' 8 6: 8 31136 31121 32134129 41:39 40:38 38:36 30136.38 38:34 6:21119:11 16110'10 8: 1 -8.04 -8.13 Unequal T Test only run where F Test Failed 13.00 12.00 3,11 -0.19 0.18 14.00 12.00 15.00 1,06 12.00 2.53 15.00 1.10 12.00 �0 3.08 14.001 -0.19 11.00 0.05 12,00 0,21 13,00 1,61 12.00 Unequal T Test only run vhere F Test Failed -4.89 -14.08 -14.41 -3,37 Unequal T Test only run where F Test Failed 14.00 14.00 12.00 14,00 -0.06 0.00 1.99 -0.06 Unequal T Test only run vhere F Test Failed 12.00: 11.00: 13,00 14.00 0.51 13,00 0,60 11.00 0.64 11.00 2.19 14.00 3.05 13.00 0.59 12.00 0.89 13,00 1.01 15,00 2,92 * 13,00 Unequal T Test only run vhere F Test Failed Unequal T Test only run vhere F Test Failed -10.84 12.00 -10.93 11.00 -1.11 13,00 -10.40 :; 14.00 Unequal T Test only run vhere F Test Failed -21.51 -6.28 Unequal T Test only run vhere F Test Failed 14.00 14.00 14,00 1.85 2.46 2,53 2.90 3.21 Unequal T Test only run Where F Test Failed 1 Saaple 1.D.Ie Test Date Control Group Reproduction 11 2 3 4 5 6; 1 8 9;10 11 12 Vilcoxon Test Test A vs Control CC CT Value C Val Vilcoxon Test Test B vs Control CC CT Value C Val Vilcoxon Test Test C vs Control CC CT Value C Val Vilcoxon Test Test D vs Control CC CT Value C Val Vilcoxon Test Test E vs Control CC CT Value C Val Vilcoxon Test Test F vs Control CC CT Value C Val Vilcoxon Test Test G vs Control CC CT Value C Val 1-13 1-14 1-16 1-11 1-18 :-l9 -20 -22 -24 I-1 II-1 v-1 V-2 V-3 V-4 V-5 V-6 V-I V-2 V-3 VI -I V1-2 VI-3 I-25 I-26 06/14/88 26; 21 07/15/88 45:41 08/21/88 21;11 10/03/89 24:29 10/17/88 31119 10/25188 11 8 03/13/89 2110 04/24/89 31 32 05/08/89 8 24 05/09189 28 22 05/09/89 2410 07/17/89 29131 08/01189 45145 0102/89 24:28 0/17/89 24;27 0/17/89 21;34 0/18/89 :38:33 1106189 ; 35; 28 1/21/89 '29::30 2/13/89 142129 2/27/89 131;29 2120/11/89 8 :19113 0/23/89 15;15 0/03188 21; 24 1/15/88 11 25 09/11189 30 35 0/12/89 34 32 0/16/89 36 32 0/23/89 29 33 0/23189 2124 2/06189 26 22 2/13/89 18 26 06108/88 22 22 03114/89 21 24 10/16/89 24 0 01108/90 36 31 01 /08190 41 41 34 32 34 28;21 36 42 34 46 40 16 19 15 19 10 30 29 26 28 29 1620161216 1335193412 32 31,35 31 30 24; 9 18 25 27 34124 31 3I 14 13 :13 23 12 24 29' 34 21 33 41 40 46 43 41 28 28 30:21 31 302724;3326 30 31 31:31 32 41 32 37:33 31 32 33:33'31 22 18 �32'32 32 33 11 19140 34138 26; 29127 30136 31,21131133:31 1III17110 23 22 31 331 32' 30 37 22 20 27 23 34 41 IVI IVI111 IV 21; 3121123 21:22:20:18 13122 31128 31139 31140 39133 36133 34141 32; 38 26 32 30;37 252835131 20 11 18121 32 23 22128 19 15 22120 13 16 11118 32 29 22149 28 28 38;29 35 43 43; 39 I 22 31 11 29 12 337 26 30 23 31 40 25 21 33 29 35 151362929 31:29 42 39 11;17 11 19 32; 29 29 28 34118 31 28 10 21 18 15 20 36 31 39 29 24 23 2I 26162025 23 9 23 8 31 32 29 22 44 46 47 42 29 30 30 21 29 31 26 14 30 32 30 33 29373238 26 25 29 36 28 32 21 30 141;38118'39 5:21:21:34 35 30:30:35134 31 14126:16120 2 23: 4125123 30 20:20123' 24 18 11:24:23 18 18 35137:30 34 33 29139133 36 36 34:36:39 33 38 33; 28; 35 35 29 24:39:29 34 33 22:20:18 18 15 32121118 26 23 19120 21 21 19 25131 32 28 19 0;21 36 33 20 35136 36 31 36 38140 44 31 45 Test Group Reproductio 24; 20 23:29 28:30 14:18 6; 6 12:20 26:30 41:42 8;11 21130 11'32 36 29 14 12 16.20 19113 26; 31 38 ; 40 6; 6 33 26 26 23 4 36 38 21 29 14 33 9 18 10 31 38 19 25 27 12 0 3327 22 16 24.25 11; 6 21' 32 36 30 11 16 21 11 28 12 14 1 34 36 10 25 16 32 21 5 8 29 38 10 28 20 26 12 19 6 41 38 8 25 0 32 21 20 8 39 34 2 2 2 2 2 2 2 2 2 0 2 2 0 2 2 2 2 2 2 2 21 21 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 130,0 116,0 148,5 106' 109 109 2 2 2 2 2 2 2 2 0 2 2 0 2 2 2 2 2 2' L. 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 21 2' 1 n 2' 2; 2 2 2 2 2 2 2 2 2 2 2 2 2 2 101,5 106 2 2 2 2 2 1 2 2 2 2 2 0 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 161,5 106 2 2 2 2 2 2 2 2 2 2 21 2 ; 155.5 ; 109 I 2,: 2 1 2 1 I 2' 1 2 1 1 2 I I 2 2 2 2 2 2 2 2 112.5 109 2 2' I 21 2' 2; 2' 2 2 2 2 0 2 0 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2' 2 2 2193.5* 106 2' 21 2 ; 141,0 109 2; 2' 2' 2; 2 2 2 2 21 I 2 1 2: 2190.0=; 109 21 1 21 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 0 2 2 2 2 2; 2' 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 100.0* 95.5* 109 106 2 2 2 2 2 2 2 2 2 0 2 0 2 2 2 2' I 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 80,0 99 18.0 = 99 109.5 99 78.0 * 99 84.5 * 99 110,0 99 89.5 * 96 112,0 99 80.5 * 99 101,0 99 78.0 * 99 18,0 = 99 18.0 * 99 80,0 * 99 18.0 * 99 78,0 = 99 18,0 = 99 . 86.0 *, 9g 9 ' 18.0 : 99 110,5 99 104.0 99 83,5 * 99 90,0 * 99 78,0 * 99 18.0 * 99 18.0 * 99 18.0 : 99 18.0 * 99 89,0 * 99 82.0* 99 86.0 * 99 88.0 * 99 101.599 } 78,0 • 99 18.0 * 99 2 2 2 2 2 1 2 2 0 2 0 2 2 2 2 2 2. 2: 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 115,5 2 2 2 2 2 2 2, 2 ; 169.0 2 2 2 2 2 2 2 2 106 109 Saiple I.D. 1 I-1 I-2 I-3 1-4 I-5 I-6 I-7 I-8 I-9 I-10 I-11 I-13 I-14 1-I6 I-17 I-18 I-19 I-20 1-21 I-22 I-23 1-24 II-1 III-1 IV-1 IV-2 IV-3 IV-4 IV-5 IV-6 V-1 V-2 V-3 VI-1 vI-2 VI-3 I-25 1-26 ' Test Date 06114188 01/15/88 08/21/88 10/03/89 10111188 10/25/88 03/13/89 04/24/89 05/08/89 05/09/89 0S/09/89 01117/89 08/01189 0102/89 0111189 0111189 0/18/89 1/06/89 1121189 2113/89 22189 2/21189 0/17/88 0/23189 0/03/88 1115188 09/ 11 /89 0/12/89 0/16/89 0123/89 0/23/89 2/06/89 2/13/89 06/08/88 03/14/89 10/16/89 01/08/90 01/08/90 Control Group Reproduction 2 3 4 5 6: 1, 8 1 , 1 , 9:10 11:12 26 21 34 32 34 28:21:22 15:36 29:29 4541 36423446:40:31 31:29 42:39 21 11 16 19 15 19110117 11:17 11119 24 29 30 29 26 28129129 32:29 29128 31 19 30 11 34 20:20' 36 34:18 31:28 17 8 16,20 16 12116 12 10:21 18 15 27 10 13135 19 34' 12 31 20136 31 39 31,32 32:31 35 31 26 29'24 23 21 8' 24 30:24 9 1$ 25 13 10 23 4 21 28:2227:342431,31 3026162025 24110 14:13 13 23:12 23 23 923 8 29:31 24:29 34 21:33 31 31 32 29 22 45,4541:404643141 4044464142 24:28 28128 30 21:31 25;29 30 30 27 24,21 30'21243312621:2931 2614 21'343031 31 31:3233:30323033 3813341 323133:3141129373238 35,2832333337:2229:26252936 29'3018,323232133351283221 30 42:29:11:19' 40 34138 36141 38 18 39 31,29,26:29:21 30136 5127 27 34 35 36 34 31 21.3113313730130353431 19 13 19 16' 10:17 18 14126 16 20 2 15152327 3:212323:4252330 21 24 22 21 22:20 18 20120 23 24 18 1725191322'31 281112423:1818 303531 3l3931 403513730:3433 34 32, 33 39 33 36 33 29139 33136 36 36321323441 32383413639'3338 29331302632303733128353529 21 24:31 25 28 35 37 24139 29 34 33 26 22122 20 17 18 21 22120 18 18 15 1826:23'3223222832121182623 22 22120 19,15 22 20 19120 21 21 19 21 24121 1316 17 18 25131 32 28 19 24 0123 32129 22 49 0121 36 33 20 36 31:34 28, 28, 38 29 35136' 36 37 36 41 41141 35'43:43 39 38:40 44 31 45 24:20 23:29 28130 14:18 61 6 12120 26130 41142 Test Group Reproductio 21 11 36 14 16 19 26 38 8111, 6 30:33129125 32126114:12 29:26:33:33 121231 9122 201 4:18:24 13: :10:11 31:36:31:21 40:38:38136 6:21:19:1/ 27:21 l 25 28 0111116 20 27:28 32 26 16112121 12 25'14:519 6 1; 8 6 323412941 30 36:38 38 16 10110 8 , 1 25 0 32 21 20 8 39 34 Vilcoxon Test Test 8 vs Control CC CT Value C Val 2 2 2 2 2 2 2 2 2 0 2 2 0 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 215,0 2 214.0 2' 2 214.0 2' 2' 2 2 2' 2' 2 2 2 2 2 2 2 205,0 2 2 106 109 109 109 Vilcoxon Test Test I vs Control CC CT Value C Val 2 2 2 2 2 2 2 1 2 2 2 2 2 0 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 90.50! 119.00 90.00! 106 109 109 ' aaole. Ttst .5. # Date 1 2 Control Group Reproduction 3 4 5:6 7 8 9;10 11 12 Wilcoxoa Test Test A vs Control CC CT Value C Val Wilcoxon Test Test B vs Control CC CT Value C Val Vilcoxon Test Test C vs Control CC CT Value C Val nilcoxon Test Test D vs Control CC CT Value C Val Vilcoxon Test Test E vs Control CC CT Value C Val Vilcoxon Test Test P vs Control CC CT Value C Val Vilcoxon Test Test G vs Control CC CT Value C Val 1-27 -28 -29 .-30 :-30 -32 IV-11 v'-S Y-6 -1 V-12 VI-4 VI-5 v1-6 v1-1 vl-8 VI-9 VI-10 VI-11 VI-12 VII-1 VII-2 VII-3 V-15 V-21 A 8 Y G H 07/19/88 06/01/88 02/20/89 03/06/89 01110/89 05/08/89 04124/89 05/29189 06/19189 06/26189 08/09188 08/23/88 10123/89 12/06/89 12/13/89 11 /08189 09/04/89 08114/89 08/07/89 09/1l 89 10/09 89 11 29 89 01 10 89 10/16189 11/01/89 11/02/89 11/30/89 01/10/90 01/17/90 :21:15:20 30 27 25 14 22 8 18 16 16 25 39 12 33 24 13 16 23 26 35 23 33 30 19 20 30 33 35 26 24 13 15 33; 30 21.29 20; 31 27:20 16:16 18;15 16;16 11;11 21;24 31;25 26:22 22; 20 18 26 23; 32 35 33 42; 33 27 32 29;29 26 23 19 9 22 19;21 222 21 31;21 14 6;13 1,25'29 24 0; 23 32 13 16116 14 15 11:21 19 30 24:14 19 23 22; 13 15 23 22; 29 19; 20 42; 34 20; 3 43; 34 9;18 19;14 16115 28;34 30; 29 15;11 16;15 28; 35 11;18 23:22 34; 33 24; 24 24; 29 24:22 23;27 21 : 20 36 30 33; 24 31 30 25 25 31'28 29 31 31 30 36 38 43 40 15 19 12 19 20 8 303730303129 25 13 10 23 421 16 14 21 15 16 15 171623232429 23 38 26 36 33 24 343231352837 16 16 11 14 15 18 18 17 15 15 16 16 37; 24 39 29 34 33 21:22 20 18 18 15 28;32 27 18 26 23 1:3030382931 26:28 26 12 19 31 26; 20 23 25 21 24 24:2118212322 22:1928222629 l 9; 32 24 26 18 24 8' 16 11; 20,15 15 2314 25,2 ,20 23,21,1 ,10,23 29' 22; 49 0 : 21; 36; 33 : 20 11 15:18 15118 20:16:15 21 15118 11;11 11'16115 15 21; 32 16; 25 4130127 20 11:17 21:15 13,16:16 25 2 3 ; 2324 : 24 19' 22 : 20 24120 23;29 14 6 12 26 41 8 18 6 20 30 42 17 Test Group Reproduction 21:30 11:32 36;29 14112 16; 20 19;13 26;31 38 : 40 6; 6 33 26 26 23 4 36 38 21 29' 25:27 14 12; 0 33 33;21 9 22;16 18 24125 10 11; 6 31 21; 32 38 36; 30 19 11; l6 21 11 28 12 14 1 34 36 10 25:28:25 16; 20; 0 . 32:26' 32: 21 12 21 51920; 868; 29 41.39; 38 38; 34; 10 8; 1; 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 98.00* 109 148.50 1 109 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2' 2' 2' 2' 2' 2' 2' 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 153.50 153.00 109 109 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2' 2' 2' 2' 2' 2' 2' 2' 2' 2'- 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 120.50 ; 109 204.00 ; 109 172.50 109 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2' 2' 2' 2 2 2 90.00*; 109 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 90.00* 109 80.00 80.00* 18.00* 113.00 18.00* 112.00 100,00 92.50* 78.00* 79.00* 102.00 102.00 18.00* 89.00* 82.00* 89.Q0* 83.00* 91.50* 83.50* 81.00* 82.00* 102.00 96.50* 101.50 101.00 100.00 95.50* 82.50* 99 99 99 99 99 99 999 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 140.00 109 SaipTe ' I.D. 1 •Test Date Control Group Reproduction Vilcoxon Test Test H vs Control 1: 2: 3: 4: 5: 6: 7 IIIII 8 9 10 11:12: CC CT Value C Val Vilcoxon Test Test I vs Control CC CT Value C Val I-27 I-28 I-29 I-30 1-31 1-32 IV-1 IV-13 IV-15 IV-17 V-4 V-5 V-6 V-1 V-I2 VI-4 VI-S VI-6 VI-1 VI-8 VI-9 VI-10 YI-11 VI-12 VII-1 YII-2 VII-3 V-15 V-2I 07/19/88 30:16:23 30122'29'36 06/01/88 ' 21' 25126 33:19 20 02120189 25 39:35 35:42 34 03/06/89 14 12123 18120 3 07/10189 ,22 33133 26'43 34 05/08/89 1 8 24:30 24 9 18 04/24/89 :18 13119 13 19 14 05129/89 :16 16120 15 16 15 06/19/89 133 30121 29 28 34 06126189 '20 31:27 20, 30 29 08/09188 16 16118 15115 11 08/23/88 16 16111 17:16 15 10/23/89 27 24131 25:28 35 12/06/89 26 22:22 20:11 18 12/13/89 18 26123 32:23 22 11/08/89 35 33142 33134 33 09/04/89 21 32129 29124 24 08/14/89 26 5132 11:24 29 08/01189 23 19119 20124 22 09/11/89 9 22119 21123 27 10/09/89 22 21:31 21121 20 11/29/89 :24:13:18,141 8 07/10/89 :16: 1125129125 10/16/89 :24: 0:23' 32:29 11101/89 ,13,16'16114117 11 /02/89 15111 21 19.21 11/30/89 .30124 14 19' 15 01110/90 23122 13 15120 01/11190 21,15'20,23:25 30 33 24 31:30 25 25 31 28 29131 3730363843140 151912192018 3031303031129 25 13 10 23 4121 16 14 21 15 16:15 111623232429 23 38 26 36 33 24 34 32 31 35 28 37 16 16 11 14 15 18 18 11 15 15 16 16 372439293433 21 22 20 18 18 15 28 32 27 18 26 23 1 30 30 38 29 31 26 28 26 12 19 31 26 20 23 25 21 24 242118272322 22 19 28 22 26 29 193224261824 16117' 20:15115:23114 24:20 23121114110123 22149 0121136133120 15' 18 15118:20116115 15 18 11 11111 16 15 21321625143027 17 17 21 15113 16 16 2323124241192220 A s B 4 D 8 P G 8 Test Group Reproductio 2420213033 2329173226 28 30 36 29 26 11612024 66, 12 20 19113 26 30 26:31 36 41 42 38 : 40 38 811 61621 29:25 14:12 33 : 33 9:22 18124 10111 31121 38136 19:11 27 0 27 16 25 6 32 30 16 21 11 28 12 14 1 34 36 10 25 16 32 2� 8 29 38 10 28 20 26 12 19 6 41 38 8 25 0 32 20 8 39 34 1 , , , 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 222.00 2 2 195.50 2 221.00 2 2, 2� 2 205,00 2 2 2 109 109 109 109 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 110.00 2 2 2 2 2 90,00* 2 2 2 2' , 2' , 2' 2: 2 2 109 109