Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices 57893 clarity of the information to be They will also be asked if they have the best of the manufacturer’s collected; and (4) ways to minimize the contingency plans. The survey will also knowledge. The information will be burden of the collection of information ask if they have tested, verified, and used for possible FDA inspectional on respondents, including through the certified their systems. The request will followup, if it indicates potential unsafe use of automated collection techniques, also ask for a single point of contact at food manufacturing situations, as well when appropriate, and other forms of the manufacturer to discuss as in the preparation of industry and information technology. information. consumer directed material addressing Title: Survey of Food Manufacturing The manufacturer will provide paper Year 2000 concerns. Facilities for Year 2000 Compliance copy of the information to FDA. The Respondents: Food manufacturers. Facilities will be asked to provide a provision of information will signify FDA estimates the burden of this status on their Year 2000 readiness. that the information provided is true to collection of information as follows:

TABLE 1.ÐESTIMATED ANNUAL REPORTING BURDEN 1

Annual No. of Respondents Frequency per Total Annual Hours per Total Hours Response Responses Response

250 1 250 1 250 1 There are no capital costs or operating and maintenance costs associated with this collection of information.

FDA establishment inventory lists Drug Administration, 200 C St. SW., ACTION: Notice. were used to determine the number of Washington, DC 20204, 202–418–3091. firms who would be subject to this SUPPLEMENTARY INFORMATION: In a notice SUMMARY: The Food and Drug collection. FDA estimates that it will published in the Federal Register of Administration (FDA) is publishing two take firms an average of 2 hours to August 3, 1995 (60 FR 39753), FDA related guidance documents entitled collect, prepare, and submit the announced that a food additive petition ‘‘Guidance for Industry: Reducing requested information. (FAP 5B4467) had been filed by Ciba- Microbial Food Safety Hazards for Dated: October 20, 1999. Geigy Corp., Seven Skyline Dr., Sprouted Seeds’’ and ‘‘Guidance for William K. Hubbard, Hawthorne, NY 10532. The petition Industry: Sampling and Microbial Testing of Spent Irrigation Water During Senior Associate Commissioner for Policy, proposed to amend the food additive Planning and Legislation. regulations in § 178.2010 Antioxidants Sprout Production.’’ These guidances are intended to provide [FR Doc. 99–27977 Filed 10–26–99; 8:45 am] and/or stabilizers for polymers (21 CFR 178.2010) to provide for the safe use of recommendations to suppliers of seed BILLING CODE 4160±01±F poly[[6-[(1,1,3,3- for sprouting and sprout producers tetramethylbutyl)amino]-s-triazine-2,4- about how to reduce microbial food DEPARTMENT OF HEALTH AND diyl] [2,2,6,6-tetramethyl-4- safety hazards common to the HUMAN SERVICES piperidyl)imino] production of raw sprouts to ensure that hexamethylene[(2,2,6,6-tetramethyl-4- sprouts are not a cause of foodborne Food and Drug Administration piperidyl)imino]], as a light stabilizer in illness and to ensure that they comply polymers used as an indirect food with the food safety provisions of the [Docket No. 95F±0187] additive. Ciba Specialty Chemicals Federal Food, Drug, and Cosmetic Act Corp., has now withdrawn the petition (the act). The first guidance is based Ciba Specialty Chemicals Corp.; without prejudice to a future filing (21 largely on recommendations from the Withdrawal of Food Additive Petition CFR 171.7). National Advisory Committee for Dated: October 12, 1999. Microbiological Criteria for Food’s AGENCY: Food and Drug Administration, report entitled ‘‘Microbial Safety Alan M. Rulis, HHS. Evaluations and Recommendations on Director, Office of Premarket Approval, ACTION: Notice. Sprouted Seeds’’ (the 1999 NACMCF Center for Food Safety and Applied Nutrion. report) (Ref.1). The second guidance is SUMMARY: The Food and Drug [FR Doc. 99–27976 Filed 10–26–99; 8:45 am] intended to assist sprouters in Administration (FDA) is announcing the BILLING CODE 4160±01±F implementing one of the principle withdrawal, without prejudice to a recommendations (i.e., microbial future filing, of a food additive petition testing) in the broader sprout guidance. (FAP 5B4467) (filed by Ciba Specialty DEPARTMENT OF HEALTH AND Chemicals Corp.; formerly named Ciba- HUMAN SERVICES DATES: Written comments may be Geigy Corp.) proposing that the food submitted at any time, however, additive regulations be amended to Food and Drug Administration comments should be submitted by provide for the safe use of poly[[6- [Docket Nos. 99D±4488 and 99D±4489] December 13, 1999, to ensure adequate [(1,1,3,3-tetramethylbutyl)amino]-s- consideration in preparation of revised triazine-2,4-diyl] [2,2,6,6-tetramethyl-4- Guidance for Industry: Reducing documents, if warranted. Microbial Food Safety Hazards for piperidyl)imino] hexamethylene ADDRESSES: Submit written requests for [(2,2,6,6-tetramethyl-4-piperidyl)imino]] Sprouted Seeds and Guidance for Industry: Sampling and Microbial single copies of the guidance entitled as a light stabilizer in polymers used as ‘‘Guidance for Industry: Reducing indirect food additives. Testing of Spent Irrigation Water During Sprout Production Microbial Food Safety Hazards for FOR FURTHER INFORMATION CONTACT: Sprouted Seeds’’ and/or the guidance Julius Smith, Center for Food Safety and AGENCY: Food and Drug Administration, entitled ‘Guidance for Industry: Applied Nutrition (HFS–215), Food and HHS. Sampling and Microbial Testing of

VerDate 12-OCT-99 16:47 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00054 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm04 PsN: 27OCN1 57894 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices

Spent Irrigation Water During Sprout and have led to a significantly better disinfection treatments (such as 20,000 Production’’ to the Office of Plant and understanding of the microbial ecology parts per million of calcium Dairy Foods and Beverages (HFS–306), of sprout-associated foodborne illness, hypochlorite in water) is likely to Center for Food Safety and Applied not all industry segments have been reduce the level of contamination if Nutrition, Food and Drug reached. Traceback investigations reveal present in or on seeds and, in turn, Administration, 200 C St. SW., that most of the firms associated with reduce the risk of foodborne illness from Washington, DC 20204, 202–205–4200. recent outbreaks were not using the consumption of sprouted seed. Send one self-adhesive label to assist approved seed disinfection treatments, However, current approved treatments that office in processing your request. or were not using them consistently, do not guarantee total elimination of The guidances are attached to this and were not testing for microbial pathogens. If even a few pathogens notice as appendixes 1 and 2 and are contamination during sprout survive a seed disinfection treatment, also accessible via the FDA home page production. Although currently they can grow to high levels during on the Internet: http://www.fda.gov. approved treatments can significantly sprouting and contaminate the entire Submit written comments on the final reduce pathogen levels in or on seeds, batch. Therefore, although seed guidance(s) to the Dockets Management they have not been shown to completely disinfection treatment is recommended, Branch (HFA–305), 5630 Fishers Lane, eliminate pathogens (Ref. 1). microbial testing of spent irrigation rm. 1061, Rockville, MD 20852. Consequently, outbreaks continue to water from each batch or production lot FOR FURTHER INFORMATION CONTACT: occur. of sprouts should also be conducted to Michelle A. Smith, Center for Food On July 9, 1999, FDA issued a prevent contaminated product from Safety and Applied Nutrition (HFS– consumer advisory advising all entering the food supply. 306), Food and Drug Administration, consumers to be aware of the risks In addition, FDA is issuing ‘‘Guidance 200 C St. SW., Washington, DC 20204, associated with eating any variety of to Industry: Sampling and Microbial 202–205–2975, FAX: 202–205–4422, e- raw sprouts, and advising persons at Testing of Spent Irrigation Water During mail: [email protected]. high risk of developing serious illness Sprout Production’’ (the microbial SUPPLEMENTARY INFORMATION: due to foodborne disease (children, the testing guidance). This guidance is elderly, and persons with weakened designed to assist sprouters in designing I. Background immune systems) not to eat raw sprouts a microbial testing program to ensure Since 1995, raw sprouts have been (Ref. 3). This advisory is updated from adulterated product does not enter increasingly implicated in foodborne a previous advisory issued August 31, commerce. Specifically, this guidance outbreaks. Between January 1995 and 1998 (Ref. 4), and was prompted by recommends testing spent irrigation May 1999, there were 11 reported information from clover and alfalfa water from each individual batch or outbreaks in the United States sprout-associated salmonellosis production lot of sprouts for two associated with sprouts from outbreaks that occurred from January pathogens, E. coli O157:H7 and commercial growers, 9 of which were 1999 through May 1999. Salmonella. The microbial testing due to various Salmonella serotypes and guidance also provides instructions for II. Guidance 2 to Escherichia coli O157. The number the sampling and testing of sprouts for of culture-confirmed cases in each of In 1997, FDA asked the National those instances when it is not possible these outbreaks ranged from 8 to more Advisory Committee for Microbiological to test spent irrigation water. However, than 500, and more than 1,300 cases Criteria for Food (NACMCF) to : (1) sprouts should not be tested in lieu of have been reported overall (Ref. 1). Review the current literature on sprout- irrigation water when spent irrigation Alfalfa and clover sprouts have been associated outbreaks, (2) identify the water is available. implicated most often, but, because all organisms and production practices of Sprouters should be aware that the kinds of sprouts are produced under greatest public health concern, (3) microbial testing program described in similar conditions, all raw (uncooked) prioritize research needs, and (4) this guidance involves a number of sprouts may pose a risk. In all of the provide recommendations on hazards. Microbial test procedures reported outbreaks, the likely source of intervention and prevention strategies. should only be run by qualified the pathogen was contaminated seed. On May 28, 1999, NACMCF adopted a personnel, in a qualified independent However, in a large 1996 Salmonella report entitled ‘‘Microbial Safety laboratory that is separate from food Montevideo/Meleagridis outbreak, poor Evaluations and Recommendations on production areas. Additional criteria for sanitation and unhygienic practices at Sprouted Seeds’’ (Ref. 1). laboratories performing these tests are the sprouting facility may also have FDA is now issuing ‘‘Guidance to provided in the microbial testing contributed to the contamination of Industry: Reducing Microbial Food guidance. sprouts (Ref. 1). Safety Hazards for Sprouted Seeds’’ (the As more effective treatments or other Sprouted seeds represent a food safety sprout guidance). This guidance food safety controls are identified and problem because the conditions under incorporates some of the implemented, the current which sprouts are produced (time, recommendations made by the 1999 recommendation to test spent irrigation temperature, water activity, pH, and NACMCF report. The guidance water from each batch of sprouts nutrients) are ideal for the exponential identifies the most important steps, e.g., produced may be changed, e.g., to growth of bacteria. If bacterial pathogens use of good agricultural practices (Ref. periodic microbial testing as a tool for are present on or in the seed, sprouting 5), seed disinfection treatment, and validating the effectiveness of food conditions are likely to encourage their microbial testing, which the agency safety systems. proliferation (Ref. 2). believes should be implemented These guidance documents do not FDA and other public health officials immediately to reduce the risk of raw provide detailed information on all are working with industry to identify sprouts as a vehicle for foodborne individual steps that should be followed and implement production practices to illness and to ensure that sprouts in the production of seeds and sprouts. ensure that seed and sprouted seed are comply with the Food Safety Provision References and resources for assistance produced under conditions that are safe. of the act (21 U.S.C. 301–397). are listed at the end of this notice and While these efforts have improved food As noted in the sprout guidance, in the broader sprout guidance. Other safety awareness within the industry routine use of approved seed materials, in the form of further

VerDate 12-OCT-99 16:47 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00055 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm04 PsN: 27OCN1 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices 57895 guidance, educational videos, etc. will Microbiological Safety Evaluations and recommendations identify the be made available, as appropriate. For Recommendations on Sprouted Seeds, http:/ preventive controls that the Food and ∼ example, FDA and the California /vm.cfsan.fda.gov/ mow/sprouts2.html. Drug Administration (FDA) believes Department of Health Services, Food 2. National Advisory Committee on should be taken immediately to reduce and Drug Branch, in cooperation with Microbiological Criteria for Foods, 1999b. the risk of raw sprouts serving as a Microbiological Safety Evaluations and industry, are producing a Recommendations on Fresh Produce. Food vehicle for foodborne illness and ensure comprehensive educational video Control, 10:117–143. sprouts are not adulterated under the outlining sprout specific practices, in a 3. FDA, 1999, Press Release—Consumers food safety provisions of the Food, Drug, number of areas, including those Advised of Risks Associated With Raw and Cosmetic Act (the act). Failure to practices recommended in this Sprouts, P99–13, http://www.fda.gov/bbs/ adopt effective preventive controls can guidance. The agency expects the video topics/NEWS/NEW00684.html. be considered insanitary conditions to be available in early 2000. 4. FDA, 1998, Talk Paper—Interim which may render food injurious to The guidance documents (see Advisory on Alfalfa Sprouts, T98–47. health. Food produced under such appendixes 1 and 2) represent FDA’s 5. FDA, 1998, Guidance for Industry— conditions is adulterated under the act Guidance to Minimize Microbial Food Safety (21 U.S.C. 342(a)(4)). FDA will consider current thinking on prevention of Hazards for Fresh Fruits and Vegetables, microbial hazards in sprouted seeds. http://www.foodsafety.gov/∼dms/ enforcement actions against any party They do not create or confer any rights prodguid.html. who does not have effective preventive for or on any person and do not operate controls in place, in particular, to bind FDA or the public. An IV. Comments microbial testing. alternative approach may be used if FDA is soliciting public comments, These recommendations are based on such approach satisfies the requirement but is implementing this guidance the recommendations of the National of applicable statutes and regulations. document immediately because of Advisory Committee on Microbiological Following the recommendations in this continuing foodborne illness outbreaks Criteria for Foods (NACMCF, 1999) and guidance will not shield any person or associated with consumption of raw elaborate on Compliance Policy Guide any food from appropriate enforcement sprouts. Interested persons may, at any 7120.28 (CPG 7120.28). under the act if adulterated food is time, submit written comments on the Seed Production: Seeds for sprout distributed in interstate commerce. The guidance documents to the Dockets production should be grown under good guidances are being distributed in Management Branch (address above). agricultural practices (GAPs) in order to accordance with FDA’s policy for Level Two copies of any comments are to be minimize the likelihood that they will 1 guidance documents as set out in the submitted except that individuals may contain pathogenic bacteria. For more agency’s Good Guidance Practices, submit one copy. Comments should be information on GAPs, see FDA’s 1998 published in the Federal Register of identified with the appropriate docket ‘‘Guidance for Industry: Guide to February 27, 1997 (62 FR 8961). numbers found in brackets on each Minimize Microbial Food Safety FDA believes this guidance and guidance document. A copy of the Hazards for Fresh Fruits and current education and outreach efforts guidance documents and comments Vegetables’’. Copies of this guidance are will have a significant positive impact received may be seen in the office above available on the internet (http:// on those industry segments that still between 9 a.m. and 4 p.m., Monday www.foodsafety.gov/dms/ prodguid.html) or by calling the number need tools to make a safer product. The through Friday. agency will closely monitor the safety of listed in the references and resources at Dated: October 21, 1999. sprouts and the adoption of enhanced the end of this guidance. prevention practices as set out in this Margaret M. Dotzel, Seed Conditioning, Storage, and guidance. Acting Associate Commissioner for Policy. Transportation: Seeds that may be used Failure to adopt effective preventive The text of the guidances follows: for sprouting should be conditioned, controls can be considered insanitary GUIDANCE FOR INDUSTRY - stored, and transported in a manner that conditions which may render food minimizes the likelihood that the seeds REDUCING MICROBIAL FOOD SAFETY HAZARDS will be contaminated with pathogens. injurious to health. Food produced FOR SPROUTED SEEDS1 under such conditions is adulterated For example, seed should be stored in under the act (section 402(a)(4) (21 [Docket No. 99D±4488] closed or covered containers in a clean All parties involved in the production U.S.C. 342(a)(4))). FDA will consider dry area dedicated to seed storage. of sprouts—seed producers, seed enforcement actions against seed and Containers should be positioned off the conditioners, and distributors, and sprout producers who do not have floor and away from walls to reduce the sprout producers—should be aware that effective preventive controls in place, in possibility of contamination by rodents seeds and sprouted seeds have been particular, effective microbial testing. or other pests and to facilitate regular On December 27, 1999, FDA plans to recognized as an important cause of monitoring for pest problems. initiate a national field assignment, foodborne illness. The following Sprout Production: Sprouters should sending investigators to sprouting implement appropriate practices to 1 This guidance has been prepared by the Office ensure that sprouts are not produced in facilities to test water used to grow of Plant and Dairy Foods and Beverages in the sprouts (i.e., spent irrigation water) and violation of the act which prohibits the Center for Food Safety and Applied Nutrition at the production of food under insanitary to assess the level of adoption of Food and Drug Administration. This guidance preventive controls. represents the agency’s current thinking on conditions which may render food reducing microbial food safety hazards for sprouted injurious to health (21 U.S.C. 342(a)(4)). III. References seeds. It does not create or confer any rights for or In addition to seed treatment and testing on any person and does not operate to bind FDA The following references are on or the public. An alternative approach may be used for pathogens (see below), sprouters display in the Dockets Management if such approach satisfies the requirements of the should maintain facilities and Branch (address above) and may be seen applicable statute and regulations. Following the equipment in a condition that will recommendations in this guidance will not shield protect against contamination. Facilities by interested persons between 9 a.m. any person or any food from appropriate and 4 p.m., Monday through Friday. enforcement under the Federal Food, Drug, and with poor sanitation can significantly 1. National Advisory Committee on Cosmetic Act if adulterated food is distributed in increase the risk of contaminating Microbiological Criteria for Foods, 1999a. interstate commerce. product. Sprouters should employ good

VerDate 12-OCT-99 16:47 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00056 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm04 PsN: 27OCN1 57896 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices sanitation practices as a standard Traceback: Traceback cannot prevent GUIDANCE FOR INDUSTRY - operating procedure to maintain control a foodborne illness outbreak from SAMPLING AND MICROBIAL TESTING OF SPENT throughout all stages of sprout occurring. However, being able to trace IRRIGATION WATER production. Inadequate water quality a food back to it’s source quickly can DURING SPROUT PRODUCTION4 and poor health and hygienic practices limit the public health and economic [Docket No. 99D±4489] can all increase the risk of food impacts of an outbreak, if it occurs. Introduction becoming contaminated with pathogens. Information gained in traceback Sprouters may wish to review 21 CFR investigations may also help prevent Raw sprouts have been associated Part 110 which sets forth good future outbreaks. Sprout producers, seed with at least eleven foodborne illness manufacturing practices (GMPs) in producers, conditioners and distributors outbreaks since 1995. FDA and other manufacturing, packaging, or holding should develop and implement systems public health officials are working with human food that cover these aspects of to facilitate traceback and recalls in the industry to identify and implement food production. event of a problem. All parties should production practices that will assure Seed Treatment: Seeds for sprouting test their systems in advance of a real that seed and sprouted seed are should be treated with one or more problem. produced under safe conditions. While these efforts have improved food safety treatments (such as 20,000 ppm calcium References and resources: 2 awareness within the industry and have hypochlorite ) that have been approved 1. Food and Drug Administration. 1982. for reduction of pathogens in seeds or led to a significantly better Compliance Policy Guide Sec. 555.750 Seeds understanding of the microbial ecology sprouts. Some treatments can be applied for Sprouting Prior to Food Use, i.e., Dried at the sprouting facility while others Mung Beans, Alfalfa Seeds, etc. (CPG 7120.28 of sprout-associated foodborne illness, will have to be applied earlier in the ) can be viewed and printed from the WWW not all industry segments have been seed production process. However, at at the following address http://www.fda.gov/ reached and outbreaks continue to least one approved antimicrobial ora/compliance—ref/cpg/cpgfod/cpg555– occur. Consequently, FDA released a treatment should be applied 750.html guidance document, entitled ‘‘Guidance immediately before sprouting3. 2. Food and Drug Administration. 1998. for Industry: Reducing Microbial Food Guidance for Industry—Guide to Minimize Sprouters should carefully follow all Safety Hazards for Sprouted Seed’’ (the Microbial Food Safety Hazards for Fresh ‘‘sprout guidance’’). The sprout label directions when mixing and using Fruits and Vegetables can be viewed and guidance identifies a number of areas, antimicrobial chemicals. printed from the WWW at the following from the farm to the sprout facility, Testing for Pathogens: Because address http://www.foodsafety.gov/dms/ where FDA believes immediate steps currently approved antimicrobials have prodguid.html or may be obtained by calling should be taken to reduce the risk of not been shown to be capable of 202–401–9725. sprouts serving as a vehicle for eliminating all pathogens from seed, 3. Food and Drug Administration, 1999. foodborne illness and to ensure that sprout producers should conduct Press Release—Consumers Advised of Risks sprouts are not adulterated under the microbiological testing of spent Associated with Raw Sprouts. P99–13. http:/ Food, Drug, and Cosmetic Act (the act). irrigation water from each production /www.fda.gov/bbs/topics/NEWS/ NEW00684.html Specific recommendations in the sprout lot to ensure that contaminated product 4. FDA, 1999. ‘‘Guidance for Industry: guidance include: development and is not distributed. Because testing for Sampling and Microbial Testing of Spent implementation of good agricultural pathogens can be done with irrigation Irrigation Water During Sprout Production’’ practices and good manufacturing water as early as 48 hours into what is can be viewed and printed from the WWW practices in the production and generally a 3 to 10 day growing period, at http://vm.cfsan.fda.gov handling of seeds and sprouts, seed producers who plan accordingly can 5. National Advisory Committee on disinfection treatments, and microbial Microbiological Criteria for Foods. 1999a. obtain test results before shipping testing before product enters the food product without losing product shelf- Microbiological Safety Evaluations and Recommendations on Sprouted Seeds. http:/ supply. life. Testing, whether done by the The agency will closely monitor the producer or contracted out, should be /vm.cfsan.fda..gov/mow/sprouts2.html 6. National Advisory Committee on safety of sprouts and the adoption of done by trained personnel, in a Microbiological Criteria for Foods. 1999b. enhanced prevention practices as set out qualified laboratory, using validated Microbiological Safety Evaluations and in the sprout guidance. FDA plans to methods. Additional information on Recommendations on Fresh Produce. Food send investigators to sprouting facilities sample collection and microbial testing, Control. 10:117–143. to test water used to grow sprouts (i.e., including how to sample and test 7. Copies of Federal regulations in the spent irrigation water) and assess the sprouts when testing spent irrigation Code of Federal Regulations (CFR) may be adoption of preventive controls. Failure water is not practicable (as may be the purchased from the U.S. Government to adopt effective preventive controls case with soil-grown sprouts), can be Printing Office or by telephone at (202) 512– 1800. The CFR is also available at local can be considered insanitary conditions found in a companion guidance which may render food injurious to document referenced below. branches of U.S. Government Printing Office Bookstores. Information on location of regional branches is available on the WWW 4 This guidance has been prepared by the Office 2 In 1998, the Environmental Protection Agency at the following address: http:// of Plant and Dairy Foods and Beverages in the issued a ‘‘section 18’’ for the temporary use of Center for Food Safety and Applied Nutrition at the 20,000 ppm calcium hypochlorite to disinfect seed vm.cfsan.fda.gov/lrd/ob-reg.html Food and Drug Administration. This guidance for sprouting. In the fall of 1999, the exemption was 8. Sections of the CFR, such as 21 CFR Part represents the agency’s current thinking on renewed for another year. However, in order to 110 Current Good Manufacturing Practices in reducing microbial food safety hazards for sprouted ensure continued availability of this treatment, Manufacturing, Packing, or Holding Human seeds. It does not create or confer any rights for or registrants should be actively pursuing a full Food, can be viewed and printed from the on any person and does not operate to bind FDA registration under section 3 in 2000. WWW at the following address: http:// or the public. An alternative approach may be used 3 Antimicrobials are either pesticides chemicals www.access.gpo.gov/nara/cfr/index.html. if such approach satisfies the requirements of the or food additives, depending on where they are applicable statute and regulations. Following the used. As such their use on seeds for sprouting must Appendix 2—Guidance for Industry: recommendations in this guidance will not shield be approved by EPA or FDA. To find out what Sampling and Microbial Testing of any person or any food from appropriate antimicrobials have been approved by EPA or FDA enforcement under the Federal Food, Drug, and for use on seeds for sprouting, you can call 202– Spent Irrigation Water During Sprout Cosmetic Act if adulterated food is distributed in 418–3098. Production interstate commerce.

VerDate 12-OCT-99 16:47 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00057 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm04 PsN: 27OCN1 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices 57897 health. Food produced under such aseptically. Obviously, sample Furthermore, additional preparation conditions is adulterated under the act collection should be done on site, either (e.g., selecting representative (21 U.S.C. 342(a)(4)). FDA will consider by employees or by contract personnel. subsamples for analyses, blending or enforcement actions against any party Aseptic sampling procedures are stomaching, and allowing sprout who does not have effective preventive described below. particles to settle out) is required when controls in place, in particular, effective Testing testing sprouts. Each additional step in microbial testing. any procedure (sampling or testing) This guidance document, ‘‘Sampling FDA recommends that all testing for introduces a possibility for error. and Microbial Testing of Spent pathogens be conducted in an external, Consequently, sprouts should not be Irrigation Water During Sprout qualified, independent laboratory that tested in place of irrigation water unless Production,’’ is intended to assist should meet several key criteria. First, production methods make it impossible sprouters in implementing one of the the lab should be physically separated to test spent irrigation water. For principal recommendations in the from the food production facility to broader sprout guidance, i.e., that prevent cross-contamination from test example, spent irrigation water may not producers test spent irrigation water for materials. This is especially important be available when sprouts are grown in two pathogens (Salmonella and where the materials used in the soil. [Note: The recommendation to test Escherichia coli O157:H7) before enrichment step required before testing irrigation water does not preclude product enters commerce. Instructions and the positive controls (described adding the testing of sprouts (either are also provided for the sampling and below) can contain pathogens and if not sprouts collected during production or testing of sprouts for those instances properly handled may contaminate finished product), to a food safety plan when it is not possible to test spent sprouts. that includes testing irrigation water.] irrigation water. However, for the Second, the laboratory should be Sampling and testing steps specific to reasons discussed below, sprouts should staffed by personnel with training and sprouts are given in italics and may be not be tested in lieu of irrigation water. experience in analytical microbiology disregarded when testing spent techniques to ensure that tests are irrigation water. Why Test performed correctly and that all Sampling Plan Salmonella and Escherichia coli appropriate safety precautions, O157:H7 have been the major causes of including appropriate waste disposal, Sprouters should have a sampling sprout-associated illness outbreaks. are followed. Third, the laboratory plan in place to ensure the consistent should have appropriate resources and Seeds are the likely source of collection of samples in an appropriate a demonstrable quality management contamination in most of these manner. The following factors should be outbreaks. Routine use of approved seed system. If testing is done by the sprouter, then considered in determining when and disinfection treatments (such as 20,000 how to sample. parts per million of calcium the laboratory facilities, personnel, and hypochlorite in water) is likely to management system should also meet When to Sample reduce the level of contamination if all these criteria to ensure that testing is pathogens are present in or on seeds reliable and does not create food safety Pathogens are most likely to be and, in turn, reduce the risk of hazards. present at detectable levels at or after 48 hours from the start of the sprouting foodborne illness from the consumption Why Sample Irrigation Water of sprouted seed. However, current process. Levels will not necessarily Procedures are provided for testing approved treatments cannot guarantee increase after 48 hours and may decline spent irrigation water and sprouts. total elimination of pathogens. The slightly. Thus, collecting samples for Although each has advantages and same conditions that encourage testing can be done as early as 48 hours disadvantages, FDA is recommending germination and growth of seeds (e.g., after the start of sprouting. If seeds are testing spent irrigation water. presoaked (e.g., soaked in water for a temperature, available moisture, and Spent irrigation water that has flowed nutrients), also encourage the growth of short time and then transferred to over and through sprouts is a good growing units for sprouting), presoak bacterial pathogens. Even if only a few indicator of the types of microorganisms pathogens survive a seed disinfection time should be included in the 48 hour in the sprouts themselves and the minimum. treatment, they can grow to high levels microflora in spent irrigation water is during sprouting and contaminate the fairly uniform. Thus sampling If you are using rapid test kits, entire batch. Therefore, seed procedures for spent irrigation wate are samples may be collected as late as 48 disinfection treatments should be relatively simple. Furthermore, water hours prior to shipping and still provide combined with microbial testing to can be used directly in the test an opportunity for the sprouter to obtain ensure that pathogens are not present procedures described here. The only test results before product enters the before sprouts enter the food supply. potential disadvantage of testing spent food supply. However, early results will As additional food safety controls are irrigation water is that the level of allow a sprouter to take corrective identified and implemented, the current microorganisms recovered in irrigation actions sooner, minimizing the potential recommendation to test irrigation water water is about 1 log less than the level for a contaminated batch of sprouts to from every batch of sprouts produced in sprouts. If pathogens are present in contaminate other production batches. may be changed, e.g., to periodic sprouts at very low levels, it is possible Earlier testing (i.e., 48 hours after the microbial testing as a tool for validating that they might be missed in water but start of sprouting) will also minimize the effectiveness of food safety systems. recovered in sprouts. the time and resources spent on a batch Who Should Perform The Tests Testing the sprouts themselves has of sprouts if a presumptive positive is several significant disadvantages. First, found. If a firm’s action plan includes Sample collection multiple sprout samples must be taken running confirmatory tests on a Sample collection should be done by from different locations in the drum or presumptive positive before discarding personnel that have been trained to trays to ensure that the sample collected product, testing earlier rather than later collect representative samples is representative of the batch. allows more time to run additional tests.

VerDate 12-OCT-99 16:47 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00058 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm04 PsN: 27OCN1 57898 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices

How to Sample temperature, preferably at 0 to 4.4 °C (32 samples should be collected throughout ° Aseptic procedures are critical to to 40 F). Sealed coolant packs should the entire production lot (e.g., if there avoid contaminating the sample during be used to avoid contamination from are 20 trays in a production lot, collect sample collection, storing the sample(s), melting ice. samples from every other tray in the rack moving from top to bottom, side to and transporting the sample(s) to the What to Sample and How Much to lab. Aseptic sampling procedures, as Collect side, and front to back). Samples should described below, should be part of a be placed directly into a clean, sterile, FDA recommends that a sprouter test prelabeled container. firm’s plan for sample collection. for pathogens by collecting a sample of Equipment used to collect samples spent irrigation water from each 2. Sample Collection for Sprouts should be clean and sterile. Sampling production lot or batch. For purposes of If testing sprouts, thirty-two (32) tools and sample containers may be this guidance, a production lot or batch sample units should be aseptically purchased pre-sterilized. Alternatively, is defined as sprouts from a single lot of collected, approximately 50 grams each, tools and containers may be sterilized at seed that were started at the same time from different locations in the drum or 121 °C (250 °F) for 30 minutes in an in a single growing unit (i.e., a single growing trays. The total sprout sample autoclave prior to use. Heat-resistant, drum or rack of trays). Pooling samples will be approximately 1,600 g (about dry materials may be sterilized in a dry- should be avoided as pooling from 56.48 ounces or 3.53 pounds per heat oven at 140 °C (284 °F) for 3 hours. different production batches may production lot or batch). Sample units The type of sample containers used decrease the sensitivity of the tests by should be collected throughout the will depend on the type of samples diluting the level of pathogens in a entire production lot (e.g., from top to collected but may include pre-sterilized contaminated sample with samples that bottom, side to side, and front to back plastic bags, tubes, cups, and flasks. are not contaminated. Pooling samples of the drum or trays). Each 50 gram Containers should be dry, leak-proof, from different batches also complicates sample unit should be placed directly wide mouthed, and of a size suitable for the interpretation of results. If a into individual clean, sterile, prelabeled the samples. Sample containers should presumptive positive is found, the containers. (Keeping the thirty-two be properly labeled prior to starting sprouter should discard all lots sample units separate will make it easier sample collection. represented by the pooled sample or Sample collectors should wear a clean for the lab to select representative perform additional tests to determine analytical units for microbial analysis lab coat, sterile gloves, and a hair net to which batch(s) are contaminated. insure they do not contaminate the compared to pulling analytical units samples. Hands should be washed 1. Sample Collection for Spent Irrigation from a single 1,600 gram mass of immediately before sampling, and prior Water sprouts.) to putting on sterile gloves. Sterile The volumes given below for spent Microbial Testing gloves should be put on in a manner irrigation water (or sprouts) represent a Testing Procedures that does not contaminate the outside of sufficient sample size to test for both the glove. Gloves should be properly Salmonella and Escherichia coli The testing procedures described in disposed of after use. O157:H7. this guidance are screening tests. They Hands should be kept away from If testing spent irrigation water, 1 liter were chosen to obtain results as simply mouth, nose, eyes, and face while of water (about 2 pints or one quart) and quickly as possible (i.e., to provide collecting samples. should be aseptically collected as the answers in 48 hours or less) on the Sampling instruments should be water leaves a drum or trays during the presence or absence of two major protected from contamination at all irrigation cycle. pathogenic bacteria, i.e., Salmonella and times before and during use. Sampling If sprouts are grown in drums, a single Escherichia coli O157:H7. Formal instruments and samples moved 1 liter sample may be collected. confirmation methods, which take between the sampling site and the If sprouts are grown in trays, and all longer than 48 hours, are described in sample container, should not be passed trays in a production lot have a common the FDA Bacteriological Analytical over the remaining pre-sterilized trough for collecting spent irrigation Manual (published by AOAC instruments. water, a 1 liter sample may be collected International, Gaithersburg, MD). The sterile sample container should at that point. If there is no common The kits identified in this guidance be opened only sufficiently to admit the collection point for water from trays, it are AOAC approved screening tests and/ sample, place the sample directly in the may be necessary to collect water or FDA has experience with their use. container, then immediately closed and samples from individual trays and pool These are also the tests that FDA plans sealed. If collecting samples in a these samples. In this case, a sampling to use as screening tests to monitor container with a lid, the lid and plan should be devised to ensure spent irrigation water at sprouting container should be held in one hand collection of a sample that is facilities. If screening methods, other while collecting the sample. The lid representative of the production lot. than those described here are used, they should NOT be completely removed. When 10 or fewer trays make up a should first be validated either by (The lid should not be held separately production lot, approximately equal formal collaborative studies or by or placed on a counter). volumes of water should be collected comparative studies with standard The sample container should be filled from each of the 10 trays to make a total methods using the specific commodity no more than 3/4 full to prevent sample volume of 1 liter. For example, in question spent irrigation water or overflow. The air from the container collect about 100 ml of water from each sprouts. should not be expelled when sealing, of 10 trays to make a 1 liter sample; Procedures for determining the particularly for plastic bags. Samples or about 125 ml from each of 8 trays; 167 presence or absence of Escherichia coli sampling equipment should not be ml from each of 6 trays, and so on. O157:H7 and Salmonella species using exposed to unfiltered air currents. When more than 10 trays make up a the test kits listed below are provided at Samples should be delivered to the production lot, ten samples should be the end of this guidance. These laboratory promptly. Perishable material aseptically collected, approximately 100 procedures should be performed should be kept at an appropriate ml each from different trays. Again, separate from the food production

VerDate 12-OCT-99 16:47 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00059 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm04 PsN: 27OCN1 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices 57899 facility by a qualified laboratory, Dividing Samples into Subsamples for found so that, if one does occur, preferably an independent, certified lab. Analyses corrective actions can be taken quickly. The rapid test procedures described Spent Irrigation Water - A total of 1 L The following should be included in a in this guidance all involve an of spent irrigation should be collected corrective action plan. enrichment step to encourage the for analysis. Two (2) 100 ml subsamples Seed is the likely source of selective growth of pathogens, if they should be analyzed for the presence of contamination in sprout-associated are present, in order to make their E. coli O157:H7. Two (2) 375 ml foodborne illness outbreaks. Further, detection possible. These test kits will subsamples should be analyzed for the recent outbreak investigations have NOT detect pathogens in irrigation presence of Salmonella. Any unused shown that a single contaminated seed water or sprouts if the enrichment step portion of the spent irrigation water lot can result in contamination of is not performed. should be stored under refrigeration multiple production lots of sprouts. In addition, seasonal or regional pending completion of the analysis. Therefore, when a batch of sprouts is differences in water quality, type of seed Sprouts - Thirty-two (32) 50 g analytical considered to be contaminated with a being sprouted, individual sprout units of sprouts should be collected for pathogen, the batch of sprouts, the seed production factors, and variations in analysis. Two (2) of the 50 g analytical lot used to produce the sprouts, and any sampling and analytical conditions may units (25 g subsamples from each) other sprout production lots that were all impact on the effectiveness of the should be analyzed for the presence of made from the same seed lot and that screening tests. Therefore, the lab E. coli O157:H7 and thirty (30) of the 50 are still under control of the sprouter, should periodically run positive g sample units (25 g subsamples from should be discarded. controls (i.e., sprout or water samples to each) should be analyzed for the In addition, anything in the sprouting which a known quantity of pathogens presence of Salmonella. Unused facility that has come into contact with have been added) to ensure the tests portions of the sprout analytical units the contaminated production lot or its used are capable of detecting pathogens should be stored under refrigeration water (e.g., drums, trays, bins, buckets, when they are present in the samples pending completion of the analysis. tool and other sprouting equipment, Sample preparation (stomaching being tested. testing equipment, and other possible sprouts) surfaces, such as floors, drains, walls, Test Kits The procedures in this guidance use a and tables), should be thoroughly blender to prepare sprouts for testing. Escherichia coli O157:H7 cleaned and then sanitized to avoid As an alternative to blending, sprouts 1. VIP EHEC, Biocontrol, Inc., contamination of subsequent batches of may be homogenized in a Stomacher Bellview, WA., (AOAC Official method sprouts. Care must be taken in handling (Model 400). To use a Stomacher, place ι 996.09) contaminated sprouts or water, 25 grams of sprouts in a sterile equipment, and test materials to avoid or Stomacher bag, add 225 ml enrichment accidental exposure to the pathogen(s). 2. Reveal E. coli O157:H7, Neogen broth and process on medium speed for Corp., Lansing, MI. 2 minutes. A) Procedure for the Escherichia coli O157:H7 Rapid Analysis of Spent Salmonella Interpretation of Results and Irrigation Water or Sprouts. 1. Assurance Gold Salmonella EIA, Subsequent Actions (AOAC Official method ι 999.08) I. Test Kits choose one.• VIP EHEC5, Interpreting Results or Biocontrol, Inc., Bellview, WA., or • 2. Visual Immunoprecipitate (VIP) Analyses should be performed in Reveal E. coli O157:H75, Neogen Corp., Assay for Salmonella, (AOAC Official duplicate (two tests for each of the two Lansing, MI. pathogens). When results are negative method # 999.09) II. Equipment and Materials (Both kits are manufactured by for all tests, results are assumed to be BioControl Systems, Inc., 12822 SE correct. When results are positive for 1. 1 Mechanical blender (capable of 32nd Street, Bellevue, WA 98005). one or both tests for either pathogen, the 10,000 to 12,000 rpm) or Stomacher results are considered presumptive and Model 400 (with required stomacher General Laboratory Instructions the grower should either: bags) Prepared Media Storage 1. Consider the presumptive positive 2. Sterile blender jars, with cover, result(s) to be true and take immediate resistant to autoclaving for 60 min at Unless noted otherwise most media can corrective actions, as described below, 121 °C be made in advance and stored at 20– 3. 1 Balance, with weights (2000 g ° ° OR 30 C (68–86 F) in the dark with a shelf 2. Ask the testing laboratory to run capacity, sensitivity of 0.1 g) life of at least one month. Media should confirmatory tests and destroy the batch 4. 1 L Erlenmeyer flask be well wrapped or contained in order only if the confirmatory tests are also 5. 2 Sterile graduated pipettes, 1.0 and to reduce evaporation. positive for the presence of a pathogen. 10.0 ml and pipette aids Equipment Sterilization In considering the second option, 6. Sterile instruments for use in taking Safe and proper operation of sterilizing remember that confirmatory testing and handling of samples (such as autoclaves requires specially trained takes extra time and will lessen the knives, tongs, scissors, spoons, etc.) personnel. The sterilization time is marketable shelf life of the sprouts. (All 7. Sterile culture tubes, 16 x 150mm or typically 121 °C (250 °F) for 15 minutes. product should be held until test results 20 x 150mm Media and Equipment Decontamination are available.) Rapid test kits are for 8. Incubator/shaker platform, 35 + 1 °C Used culture media and test kits should screening ONLY. Confirmatory testing 9. pH meter or test strips be decontaminated by autoclaving should be done using standard methods 10. Fisher or Bunsen burner before disposal. Decontamination in the FDA Bacteriological Analytical 11. Magnetic stirrer and stir bars should be performed in an area that is Manual (Edition 8, Revision A–1998). totally separated from media 5 The enrichment procedure described in this Corrective Actions guidance for the tests for Escherichia coli O157:H7 preparation and sterilization. Trained have been modified by FDA to enhance the ability personnel should be used to properly Each facility should have a corrective of the kits to detect Escherichia coli O157:H7 in decontaminate used media. action plan in place before a positive is spent irrigation water and sprouts.

VerDate 12-OCT-99 16:47 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00060 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm04 PsN: 27OCN1 57900 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices

12. Sterile syringes 10. Vancomycin (antibiotic) cool, add antibiotic supplements. 13. Sterile 0.2 m filters Instructions for sprouts are given in Preparation of antibiotic stock solutions 14. Distilled water italics. Prepare a stock solution of each III. Ingredients antibiotic (acriflavin, cefsulodin, and Modified Buffered Peptone Water 1. Peptone vancomycin) by dissolving 1000 mg of (mBPW) 2. NaCl each antibiotic in a separate tube Step 1. To make 1000 ml of mBPW, mix 3. Na2HPO4 containing 10.0 ml of distilled water. the following constituents into distilled 4. KH2PO4 Filter-sterilize the solution using a 0.2 m water, stirring to dissolve. For spent 5. Casamino acid filter and syringe. The stock solution irrigation water, prepare double strength 6. Yeast extract may be stored for several months in foil (2X) mBPW, as follows: (If testing 7. Lactose wrapped tubes at 4 C (39.2 C). sprouts, use single strength enrichment 8. Acriflavin (antibiotic) Prepare the modified Buffered Peptone broth base.) 9. Cefsulodin (antibiotic) Water as described below, autoclave,

Modified Buffered Peptone Water (mBPW) Double strength (2X) (For use Single strength Ingredient with spent irri- (1X) (For use gation water) with sprouts)

Peptone 20.0 g 10.0 g NaCl 10.0 g 5.0 g Na2HPO4 7.2 g 3.6 g KH2PO4 3.0 g 1.5 g Casamino acid 10.0 g 5.0 g Yeast extract 12.0 g 6.0 g Lactose 20.0 g 10.0 g Distilled water* 1000 ml 1000 ml *pH 7.2 ± 0.2 (Test pH of distilled water BEFORE adding the ingredients above. If necessary, pH may be adjusted with 1N HCl or 1N NaOH.

Step 2. Sterilize mBPW by autoclaving subsample, add the following filter- sprouts, add the quantity of antibiotics at 121 °C (250 °F) for 15 minutes. sterilized antibiotics to 1000 ml of listed in the column labeled single Remove from autoclave and allow to medium. For spent irrigation water, add strength to the single strength mBPW.) cool until cool to the touch. the quantity of antibiotics listed in the Step 3. Once the medium is cooled and column labeled double strength to the immediately prior to the addition of a double strength (2X) mBPW. (If testing

Antibiotic supplements for mBPW Double strength (2X) (For use Antibiotic Stock Solution with spent irri- Single strength (1X) (For use with sprouts) gation water)

Acriflavin (A) 0.2 ml 0.1 ml Cefsulodin (C) 0.2 ml 0.1 ml Vancomycin (V) 0.16 ml 0.08 ml

IV. Testing - Perform analysis in the 1000 ml sample of spent sprout mBPW+ACV) and blend at 10,000 to duplicate: For microbial testing, irrigation water, aseptically transfer 100 12,000 rpm until homogenized (at least duplicate sub-samples (analytical units) ml of sample into a sterile 1L flask 60 seconds) or stomach for 2 minutes on need to be removed from the sample containing 100 ml of 2X mBPW+ACV. medium setting in a Stomacher Model and placed in enrichment broth. Repeat with second subsample. 400. Transfer sprout homogenate to a 1L Enrichment broth containing sub- Sprouts: Two (2) 50 g analytical units of Erlenmeyer flask. samples are allowed to incubate for a sprouts will be analyzed. From two of period of time, and a small quantity of the thirty-two 50 g analytical units the enrichment broth/sample is applied collected, aseptically remove and weigh to the test kit device. Specific directions out a 25g subsample of sprouts. Transfer follow: each of the 25 gram subsamples of Step 4. sprouts into separate sterile blender jars or sterile stomacher bags. Add 225 ml of Water: Two (2) 100 ml subsamples of single strength enrichment broth with spent irrigation will be analyzed. From added antibiotic supplements (1X

VerDate 12-OCT-99 17:12 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00061 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm08 PsN: 27OCN1 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices 57901

Step 5. Incubate the enrichment broth/ 10. Sterile pipettes, 1 ml, with 0.01 ml 2. Buffered Peptone Water + novobiocin sample mixtures overnight at 37 °C (98.6 graduations; 5 ml with 0.1 ml (BPW + n) ° F) with shaking at 140 RPM. graduations and 10 ml with 0.1 ml Immediately prior to the addition of a Step 6. Test each enrichment broth graduations and pipette aids 25 g subsample, add 4 ml of 0.1% 11. Inoculating needle and inoculating sample for the presence of E. coli novobiocin solution to each 225 ml loop (about 3 mm id or 10 l), nichrome, O157:H7, using either the VIP EHEC volume of buffered peptone water. device or the Reveal E. coli O157:H7 platinum-iridium, chromel wire, or 6. Trypticase soy broth + novobiocin device. Use 0.1 ml from the inoculated sterile plastic 12. Sterile test or culture tubes, sizes 16 (TSB+n) and incubated mBPW +ACV to x 150 mm and 20 x 150 mm inoculate VIP or 0.12 ml for the Reveal. Suspend 30 g of commercial available 13. Test or culture tube racks Follow the manufacturers instructions trypticase soy broth medium in 1 L of 14. Vortex mixer distilled water. Mix thoroughly. Warm for the inoculation of test kits. 15. Sterile shears, large scissors, scalpel, gently on a temperature controlled hot Step 7. Observe test results within 10 and forceps plate until the medium is dissolved. minutes to avoid possible fading of 16. Fisher or Bunsen burner Dispense in 10 ml aliquots in 20 x 150 bands which could lead to false negative 17. pH test paper (pH range 6–8) with mm tubes and autoclave 15 min. at 121 results. A band in both the test and maximum graduations of 0.4 pH units °C. control chambers is a positive test for per color change contamination. A band in only the 18. Sterile syringe Just prior to sample addition, add 0.1 ml control chamber is a negative test. If a 19. Sterile 0.2 m filters of 0.1% novobiocin solution to each tube containing 10 ml of Trypticase soy band does not appear in the control III. Media and reagents chamber, the test was not done correctly broth. and must be repeated. For preparation of media and reagents, 7. Trypticase soy broth + 2, 4 refer to sections 967.25 to 967.28 in B) Procedure for the Salmonella Rapid dinitrophenol + novobiocin Official Methods of Analysis (published (TSB+DNP+n) Analysis of Spent Irrigation Water (or by AOAC International, Gaithersburg, Sprouts) MD USA). Designations within Suspend 30 g of commercial available I. Test kits choose one parentheses refer to Appendix 3, Media trypticase soy broth medium and 0.1 g • and Reagents, of the Bacteriological of 2, 4 dinitrophenol in 1 L of distilled Assurance Gold Salmonella EIA, or water. Mix thoroughly. Warm gently on • Visual Immunoprecipitate (VIP) Analytical Manual (BAM), Edition 8, Revision A ( also published by AOAC a temperature controlled hot plate until Assay for Salmonella the medium is dissolved. Dispense in 10 Both are manufactured by BioControl International). ml aliquots in 20 x 150 mm tubes and Systems, Inc., (12822 SE 32nd Street, 1. Buffered peptone water autoclave 15 min. at 121 °C (250 °F). Bellevue, WA 98005). For purposes of (commercially available-Oxoid, BBL, or Just prior to sample addition, add 0.1 ml pre-enrichment and selective Difco) of 0.1% novobiocin solution to each enrichment, these test kits provide 2. Buffered peptone water + novobiocin tube containing 10 ml of Trypticase soy different instructions for each of three 3. Tetrathionate (TT) broth (M145) broth + 2, 4 dinitriphenol. types of foods: (a) processed foods, (b) 4. Rappaport-Vassiliadis (RV) medium dried powder processed foods, and (c) (M132) 9. Novobiocin solution, 0.1% raw foods. For the analysis of sprouts 5. Trypticase soy broth (commercially available) Novobiocin, sodium salt 0.1 g and spent irrigation water, use the pre- Distilled water 100 ml enrichment/selective enrichment 6. Trypticase soy broth + novobiocin Dissolve novobiocin in distilled water. procedures described for (c) raw foods. 7. Trypticase soy broth + 2, 4 dinitrophenol + novobiocin Do not autoclave. Sterilize by filtering II. Equipment and materials1. Blender 8. 1 N Sodium hydroxide solution through a 0.2 m filter. Store solution at and sterile blender jars OR Stomacher (NaOH) (R73) 4 °C (39.2 °F), protected from light (e.g. Model 400 with appropriate stomacher 9. 1 N Hydrochloric acid (HCl) (R36) wrap container in aluminum foil). bags. 10. Novobiocin solution, 0.1% Solution can be stored for one week. 2. Sterile, 16 oz (500 ml) wide-mouth, 11. Sterile distilled water IV.Testing screw-cap jars, sterile 500 ml Buffered peptone water (Number 1.), A. Irrigation water-From the 1 L spent Erlenmeyer flasks, sterile 250 ml Buffered peptone water with novobiocin irrigation water sample, two (2) 375 ml beakers, sterile glass or paper funnels of (Number 2), Trypticase soy broth with subsamples will be analyzed for the appropriate size, and, optionally, novobiocin (Number 6) and Trypticase presence of Salmonella. containers of appropriate capacity to soy broth with 2,4 dinitrophenol and accommodate composited samples novobiocin (Number 7), are not 1. Aseptically transfer a 375 ml 3. Balance, with weights; (2000 g included in the BAM. Their preparation subsample directly to a 6 L Erlenmeyer capacity, sensitivity of 0.1 g) is described below. flask containing 3,375 ml BPW + n. Swirl to mix thoroughly. Repeat 4. Balance, with weights; (120 g 1. Buffered Peptone Water (BPW) capacity, sensitivity of 5 mg) procedure with second 375 ml 5. Incubator, 35 °C (95 °F) Dissolve 20 grams of commercially subsample of spent irrigation water. 6. Refrigerated incubator or laboratory available buffered peptone water 2. Allow flasks to stand for 60 min at refrigerator, 4 + 1 °C (39 + 1 °F) medium in 1 liter distilled water. Mix room temperature. Mix well and 7. Water bath, 42 + 0.2 °C (107.6 + 0.2 thoroughly. Dispense 225 ml portions determine pH with test paper. Adjust °F) into 500 ml Erlenmeyer flasks. After pH, if necessary, to 6.8 + 0.2 with sterile 8. Sterile spoons or other appropriate autoclaving for 15 min at 121 °C, and 1 N NaOH or 1 N HCL. instruments for transferring food just before use, aseptically adjust 3. Incubate flasks without shaking for samples volume to 225 ml with sterile distilled 18–26 hours at 35–37 °C (95–98.6 °F). 9. Sterile culture dishes, size 15 x 100 water. Adjust pH, if necessary, to 7.2 + Each flask is considered to contain pre- mm, glass or plastic 0.2 with sterile 1 N NaOH or 1 N HCl. enrichment broth.

VerDate 12-OCT-99 17:12 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00062 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm08 PsN: 27OCN1 57902 Federal Register / Vol. 64, No. 207 / Wednesday, October 27, 1999 / Notices

4a. If using the Assurance Gold 3. Incubate flasks without shaking for necessity and utility of the proposed Salmonella Enzyme Immunoassay, 18–26 hours at 35–37 °C (95–98.6 °F). information collection for the proper transfer 0.1 ml pre-enrichment broth to Each flask is considered to contain pre- performance of the agency’s functions; 10 ml RV medium and transfer another enrichment broth. (2) the accuracy of the estimated 1.0 ml of pre-enrichment broth to 10 ml 4a. If using the Assurance Gold burden; (3) ways to enhance the quality, TT broth. Incubate in a water bath 5–8 Salmonella Enzyme Immunoassay, utility, and clarity of the information to ° ° hours at 42 C (107.6 F). Incubation of transfer 0.1 ml pre-enrichment broth to be collected; and (4) the use of the RV medium and TT broth in the 10 ml RV medium and transfer another automated collection techniques or water bath is termed the selective 1.0 ml of pre-enrichment broth to 10 ml other forms of information technology to TT broth. Incubate in a water bath 5–8 enrichment process. Following selective minimize the information collection hours at 42 °C (107.6 °F). Incubation of enrichment, transfer and combine 1.0 burden. ml TT broth and 0.5 ml RV medium into the RV medium and TT broth in the a single tube containing 10 ml of water bath is termed the selective Type of Information Collection prewarmed [42 °C (107.6 °F)] TSB + n enrichment process. Following selective Request: New; Title of Information broth. Incubate in a water bath 16–20 enrichment, transfer and combine 1.0 Collection: Evaluation of New Medicare hours at 42 °C (107.6 °F). Continue as ml TT broth and 0.5 ml RV medium into Members of Medicare+Choice Plans; described in this kit’s instructions for (c) a single tube containing 10 ml of Form No.: HCFA–R–0298 (OMB# 0938– raw foods. prewarmed [42 °C (107.6 °F)] TSB + n New); Use: The objective of this survey 4b. If using the VIP Assay for broth. Incubate in a water bath 16–20 is to understand the special information Salmonella, transfer 0.1 ml pre- hours at 42 °C (107.6 °F). Continue as needs of new Medicare members, their enrichment broth to 10 ml RV medium described in this kit’s instructions for (c) sources of information, their preferred and transfer another 1.0 ml of pre- raw foods. distribution channels, their enrichment broth to 10 ml TT broth. 4b. If using the VIP Assay for understanding of the traditional Incubate in a water bath 18–24 hours at Salmonella, transfer 0.1 ml pre- Medicare program and their 42 °C. Incubation of the RV medium and enrichment broth to 10 ml RV medium understanding of their particular and transfer another 1.0 ml of pre- TT broth in the water bath is termed the +Choice plan, and the impact National enrichment broth to 10 ml TT broth. selective enrichment process. Following Medicare Education Program activities Incubate in a water bath 18 24 hours at selective enrichment, transfer and may have on new members’ decisions to 42 °C. Incubation of the RV medium and combine 0.5 ml of TT broth and 0.5 ml choose a +Choice plan or change their RV medium into a single tube TT broth in the water bath is termed the ° selective enrichment process. Following plan; Frequency: On occasion; Affected containing 10 ml prewarmed [42 C Public: Individuals; Number of (107.6 °F)] TSB+DNP+n broth. Incubate selective enrichment, transfer and Respondents: 3000; Total Annual in a water bath 5–8 hours at 42 °C (107.6 combine 0.5 ml of TT broth and 0.5 ml Responses: 3000; Total Annual Hours: °F). Continue as described in this kit’s RV medium into a single tube instructions for (c) raw foods. containing 10 ml prewarmed [42 °C 1212. ° B. Sprouts - Thirty 50 g analytical units (107.6 F)] TSB+DNP+n broth. Incubate To obtain copies of the supporting ° of sprouts were collected for Salmonella in a water bath 5–8 hours at 42 C (107.6 statement and any related forms for the ° analysis. Aseptically weigh out a 25 g F). Continue as described in this kit’s proposed paperwork collections subsample from each analytical unit and instructions for (c) raw foods. referenced above, access HCFA’s Web transfer each subsample to a sterile [FR Doc. 99–28016 Filed 10–25–99; 8:45 am] Site address at http://www.hcfa.gov/ blending jar (or stomacher bag). Add BILLING CODE 4160±01±F regs/prdact95.htm, or E-mail your 225 ml buffered peptone water plus request, including your address, phone novobiocin (BPW + n). Blend the 25 g number, OMB number, and HCFA DEPARTMENT OF HEALTH AND sprout subsample with 225 ml BPW + document identifier, to HUMAN SERVICES n for 2 min. Repeat procedure for [email protected], or call the Reports remaining twenty-nine analytical units. Health Care Financing Administration Clearance Office on (410) 786–1326. 1. The thirty 25 g sprout subsamples Written comments and may be analyzed by either of the [Document Identifier: HCFA±R±0298] recommendations for the proposed following two options: information collections must be mailed Agency Information Collection Option A: within 60 days of this notice directly to Each 25 g/225 ml blended sprout Activities: Proposed Collection; Comment Request the HCFA Paperwork Clearance Officer homogenate is poured into a 500 ml designated at the following address: Erlenmeyer flask, or equivalent AGENCY: Health Care Financing HCFA, Office of Information Services, container, and analyzed individually. Administration, HHS. Security and Standards Group, Division Option B: In compliance with the requirement of HCFA Enterprise Standards, Fifteen of the thirty 25 g/225 ml of section 3506(c)(2)(A) of the Attention: Julie Brown, Room N2–14– blended sprout homogenates are poured Paperwork Reduction Act of 1995, the 26, 7500 Security Boulevard, Baltimore, into a 6 L Erlenmeyer flask, and Health Care Financing Administration Maryland 21244–1850. analyzed collectively. Repeat with the (HCFA), Department of Health and remaining 15 blended sprout Human Services, is publishing the Dated: October 18, 1999. homogenates . Thus, each sample following summary of proposed John Parmigiani, consists of two 375-g composites . collections for public comment. Acting, HCFA Reports Clearance Officer, 2. Allow flasks to stand for 60 min at Interested persons are invited to send HCFA Office of Information Services, Security room temperature. Mix well and comments regarding this burden and Standards Group, Division of HCFA determine pH with test paper. Adjust estimate or any other aspect of this Enterprise Standards. pH, if necessary, to 6.8 + 0.2 with sterile collection of information, including any [FR Doc. 99–27986 Filed 10–26–99; 8:45 am] 1 N NaOH or 1 N HCL. of the following subjects: (1) The BILLING CODE 4120±03±P

VerDate 12-OCT-99 17:12 Oct 26, 1999 Jkt 190000 PO 00000 Frm 00063 Fmt 4703 Sfmt 4703 E:\FR\FM\27OCN1.XXX pfrm08 PsN: 27OCN1