Proceedings of the International Cyanide Detection Testing Workshop

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probably start with fi sh tissue or tissue homogenates. Dr. Logue pointed out, based on mammalian research, that thiocyanate can be produced from several metabolic pathways (Logue and Hinkens 2008). “However, thiocyanate is naturally found in biological fl uids, and while this is a condition of all cyanide metabolites, thiocyanate levels are normally quite high and can be inconsistent. Large variation in background thiocyanate concentrations (in mammals) make it diffi cult to determine low level cyanide exposure….Large and variable concentrations (of thiocyanate) may indicate that thiocyanate is involved in a number of biological processes in addition to cyanide metabolism.” Hence, even if thiocyanate could be detected in MAF being imported, it might not be possible to assert in court that the presence of thiocyanate detected in the fi sh was absolute proof that the fi sh were caught using cyanide in the country of origin. Based on the previous discussion, it is unlikely that a rapid fi eld test for thiocyanate can be applied. Analyses of frozen samples sent to a laboratory (such as the USFWS laboratory in Oregon) will probably be necessary. From a cost perspective, measuring thiocyanate may be much less costly than the ATCA method (discussed below). ATCA Test The committee agreed that from a theoretical standpoint is may be better to use a biomarker such as ATCA (Lindquist et al. 2005, Logue et al. 2005, Logue et al. 2007, Logue and Hinkens 2008), especially if the US-based test results will be used for prosecution of those in the aquarium trade. “A minor metabolic pathway is the conversion of cyanide ion to 2-amino- 2thiazoline-4-carboxylic acid (ATCA). Another by product (tautameric form) is 56 2-iminothiazolidine-4-carboxylic acid (ITCA). The production of ATCA may predominate when sulfur donors become depleted or in tissues where rhodanese is sparse. … ATCA may be used as an alternative to thiocyanate for determination of cyanide exposure. An advantage of using ATCA is that it is stable in biological samples for months at freezing and ambient temperatures and therefore, may be a lasting signature for cyanide exposure. However, relatively few techniques have been described to analyze ATCA from biological matrices and relatively few studies have been conducted to evaluate the relationship between ATCA and cyanide exposure” (Logue and Hinkens 2008). Nothing is known about the pharmaco-kinetics of ATCA. A lot of research is needed before it could be applied to support law enforcement. Since, Dr. Logue has done research on ATCA he may have the equipment. If the USFWS laboratory in Oregon was to adopt this test method, they would need to purchase the equipment. Lundquist et al. (1995) measured ATCA in urine using HPLC. Logue et al. (2005) measured ATCA in blood and urine using GC-MS. It is not clear how ATCA would be liberated from fi sh tissues prior to analysis. According to Dr. Logue (pers. comm. 2008) the cost of the GC-MS apparatus to measure ATCA with peripheral equipment is about $125,000. The cost of the equipment and its technical complexity most probably precludes its use for measuring ATCA in exporting countries. Validation Studies Dr. Rubec questioned the need for additional studies to evaluate the ASTM ISE methodology. There have already been several round-robin studies by the American Society of Testing and Materials (ASTM 1987, summarized ASTM 1997) and by the American Public Health Association (summarized APHA-AWWA-
WPCF 1992, 1998). Both of these studies have included at least 5 separate laboratories and 6 operators but were performed only on aqueous samples, not masticated fi sh tissue. The concerns about QA/QC, precision, and bias were addressed. The IMA also conducted round-robin comparisons between the six laboratories it ran under contract to BFAR (IMA-Philippines 1999, Rubec et al. 2003). If round-robin testing needs to be conducted, it should be done to validate a U.S.-based test for either thiocyanate or the ATCA method. It is unlikely that the ASTM ISE method will be applied in the USA. It looks like it will be used in both the Philippines and in Indonesia. An accreditation system tied to CDT analyses from export facilities may give the U.S.A. the means to prove intent. If fi sh arrive at an import facility along with an Accreditation Certifi cate, the importers would have to admit that they knew the fi sh were accredited as being cyanide-free. If a test such as the ATCA proves otherwise, there may be grounds for prosecution of the importer under the Lacey Act. Congressional legislation similar to that proposed by Congressman Ed Case of Hawaii (House Resolution 4928 for 108 th Congress in 2004) may be necessary to allow law enforcement offi cials (from USFWS and/or NOAA) to prosecute importers who knowingly import fi sh that are proven to contain cyanide. The test that would need to hold up in court would not be the ASTM ISE method. It would be either the thiocyanate test or the ATCA test conducted in the U.S.A. U.S. offi cials need to investigate the legal ramifi cations to see whether this is feasible. Funding The IMA conducted CDT in six laboratories in the Philippines under contract to BFAR for about $100,000 per year. The same testing in the U.S.A. would be at least fi ve times more 57 expensive based on the disparity of salaries between the two countries. The Philippines, Indonesia, and Vietnam need long-term funding to establish and maintain CDT networks in each country. Unlike the U.S.A., the CDT laboratories in he exporting countries should be used primarily as a deterrent to cyanide fi shing. Cyanide testing should be conducted regionally using the cyanide screening method, so that local governments or municipalities can better manage their fi sheries. When samples are found positive using the screening method, frozen specimens would be shipped to CDT laboratories established near international airports. The CDT laboratories using the ISE method need to be tied to a licensing system for fi shers and MAF collectors, middlemen, and exporters. If fi sh coming from collectors or fi shers situated in municipalities, or from exporters situated near points of export, are found to have cyanide present more than 3 or 4 times, their licenses can be revoked. Prosecution, being expensive, should only be used as a last resort. Prosecutions should probably only be conducted when either the fi shermen/collectors, the middlemen, or the exporters are apprehended with cyanide in their possession and the fi sh test positive for the presence of cyanide. It should be noted that it becomes more diffi cult to prove that fi shes have been collected with cyanide by the time the fi sh enter the U.S.A. U.S. agencies such as the U.S. Agency for International Development (USAID) or other agencies, such as the World Bank or the Asian Development Bank, should fund the creation of cyanide detection networks in countries where cyanide use is prevalent. It will be cheaper to conduct most of the CDT in the exporting countries.
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Proceedings of the International Cy
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Proceedings of the Cyanide Detectio
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ACKNOWLEDGEMENTS The Proceedings of
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PREFACE The International Cyanide D
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EXECUTIVE SUMMARY The International
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ATCA) in homogenized fi sh tissue m
Page 15 and 16: Validation of the ISE method applie
Page 17 and 18: Importing countries should also req
Page 19 and 20: INTRODUCTION Cyanide fi shing is a
Page 21 and 22: through the implementation of a net
Page 23 and 24: Working Group 2 The Export Working
Page 25 and 26: o o o o Can the U.S. require verifi
Page 27 and 28: Polymeric membrane based ion select
Page 29 and 30: Goal 2: Provide recommendations on
Page 31 and 32: hands-on training that develops a d
Page 33 and 34: Goal 3: Identify research needs to
Page 35 and 36: Working Group 1 Participants Bob KO
Page 37 and 38: estimated at roughly 3 to 5 days. H
Page 39 and 40: Goal 2 : Identify steps, agreements
Page 41 and 42: 3) certifi cate, but tests positive
Page 43 and 44: Report from Working Group 3 The Par
Page 45 and 46: No. of Shipments 6,000 5,000 4,000
Page 47 and 48: Goal 3: Provide a recommendation wi
Page 49: Working Group 3 Participants Eric P
Page 53 and 54: Introduction Since the early 1960
Page 55 and 56: ainbow trout exposed to 40 mg SCN -
Page 57 and 58: Blennies, which were net-caught and
Page 59 and 60: electrical potential (similar to an
Page 61 and 62: in the Philippines under contract w
Page 63 and 64: endorsing the IMA’s use of the IS
Page 65: The intended products from the rese
Page 69 and 70: ASTM (1997) Standard test methods f
Page 71 and 72: Kuegsen, M., Kloock, J.P., Knobbe,
Page 73 and 74: Philippines. In Proceedings from th
Page 75 and 76: 3) 4) 5) 6) the metabolism of cyani
Page 77 and 78: or new matrices. 5.2.3 Modifi catio
Page 79 and 80: Level Two: The method should be val
Page 81 and 82: of cyanide exposure and to identify
Page 83 and 84: Special considerations for the anal
Page 85 and 86: tissue samples indispensable. Altho
Page 87 and 88: y reduction and subsequent separati
Page 89 and 90: can also be used for analysis of cy
Page 91 and 92: thiocyanate in saliva and serum bas
Page 93 and 94: are many discrepancies in the liter
Page 95 and 96: Table 3. Analytical Methods to dete
Page 97 and 98: Poisoning, In Textbook of military
Page 99 and 100: (70) Odoul, M., Fouillet, B., Nouri
Page 101 and 102: V. (1975) Stable reagents for the c
Page 103 and 104: (159) Felscher, D. and Wulfmeyer, M
Page 105 and 106: Purpose of this Document The purpos
Page 107 and 108: day. One cyanide tablet (2 g) is mi
Page 109 and 110: 10-90% following exposure to cyanid
Page 111 and 112: existing cyanide detection methods
Page 113 and 114: - Plasma lactate — elevated plasm
Page 115 and 116: Method Advantages Disadvantages Hig
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Paper Method Matrix Detection Limit
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Table 4 lists experts identifi ed t
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X Jerry Thomas CDC Em Steve Borron
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References Barber, C. and Pratt, V.
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Rates of Recovery. Environmental Ma
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COUNTRY REPORTS
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The first three of these destructiv
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enforcement is minimal. There is an
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1. Management/guidance to the Minis
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in Vietnam now and requires more ef
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Figure 2: Administrative structure
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References Baquero, J. (1999) Marin
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Acronyms • CDT • Cyanide Detect
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Law/Ordinance/Decree /Decision 52/2
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of samples required for analysis. U
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Field test kit. BFAR recognizes the
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Political Will Having planted the s
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gravely by fi shing communities. Th
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ecome the partner and counterpart o
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Ion-Selective Electrodes for Cyanid
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References Alban, J., Manipula, B.E
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levels with mV readings recorded as
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Trends Determined by Cyanide Testin
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Implementing agencies: • Quaranti
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APPENDIX
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1:30 Overview of and Potential Cyan
Page 172 and 173:
Kristine Johnson Director Kingfi sh
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Assessment of U.S. Role in Trade: U
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