臺灣博碩士論文加值系統

高壓加工技術 (High pressure processing, HPP) 是應用於食品殺菌和保存的一項非熱加工技術，可保持食品的新鮮度但不會影響某些食品品質特性，例如顏色，天然風味和營養成分等。本研究首先探討高壓技術對組織胺高中毒風險魚種-虱目魚柳之殺菌與保鮮應用，以高壓200、300、400、500和600 MPa維持5分鐘處理虱目魚柳後，發現魚肉中之總生菌數 (Aerobic plate count, APC)、低溫菌數 (Psychrotrophic bacteria count, PBC)、產硫化氫菌數 (H2S-producing bacteria count, HBC)、大腸桿菌群 (Coliform) 和總揮發性鹽基態氮 (Total volatile basic nitrogen, TVBN) 的數值會隨壓力增加而降低。同時魚肉之L* (亮度)、W (白度)、ΔE (色差值)、硬度、內聚力和咀嚼力等數值會隨著壓力增加而顯著上升，反之，a*值 (紅色度) 則隨壓力增加而下降；顯示壓力愈高時魚肉的外觀顏色愈偏白與亮，同時伴隨質地變硬。此外，將前述加壓後之魚肉於4℃與15℃下進行貯存，評估魚肉中組織胺相關衛生品質之變化。發現高壓處理可延遲虱目魚柳在低溫 (4℃及15℃) 貯存時APC、PBC、TVBN 與組織胺含量之增加，並且壓力愈高延遲效果愈明顯；總而言之，若配合低溫4℃與較高壓力 (>400 MPa) 時可顯著延緩APC、PBC之生長以及TVBN與組織胺的產生，同時延長魚柳之保存期限。
另外，應用鹽水漬結合高壓處理之柵欄技術於提升虱目魚柳之品質，乃將魚柳先以3% 或 9%鹽水浸漬，再結合不同高壓 (300、400、500及600 MPa，5分鐘) 處理後，評估對魚柳之衛生品質與物理性質影響。可知以鹽水漬配合高壓處理比單獨鹽水漬或單獨高壓處理，對魚肉之APC、PBC 與TVBN數值具有明顯減少的加乘效果。再者，在相同壓力下，鹽水漬組之魚肉較為澎潤與亮白，其L*與W值均較未鹽漬者為高，並且9%高鹽漬組會更使魚肉原本紅色色澤明顯退色。其次，鹽水漬會使魚肉質地較為柔軟，可改善因高壓處理所導致魚肉硬度及咀嚼力增加 (變硬) 之缺點。因此建議虱目魚柳以3%低鹽水漬結合適度壓力 (300 或400 MPa) 處理，可維持較佳的色澤、質地與口感，以及較低微生物菌數與TVBN值。

High pressure processing (HPP) is a non-thermal technology for food pasteurization and preservation. The usage of HPP treatment to preserve the freshness of food was also shown not to affect some of the food quality characteristics such as the color, natural flavor, and nutrients. In this research, the HPP treatment (200, 300, 400, 500, and 600 MPa for 5 min) on preservation application of milkfish sticks was studied. The values of aerobic plate count (APC), psychrotrophic bacteria count (PBC), H2S-producing bacteria count (HBC), coliform and total volatile basic nitrogen (TVBN) in raw milkfish sticks after HPP treatment decreased with increased pressure. In color and texture of milkfish sticks, the values of L* (lightness)、W (whiteness), ΔE, hardness, chewiness and cohesiveness increased significantly with increasing pressure, whereas a* value (redness) significantly decreased with increasing pressure. In addition, the changes of APC, PBC, TVBN, and pH values in milkfish sticks after HPP treatment during storage at different temperatures (4℃ and 15℃) were evaluated. HPP treatments (≧400 MPa) for 5 min were significantly delayed the APC, PBC, and TVBN increase in milkfish sticks stored at low temperature (4 or 15℃), and extended the shelf-life of milkfish sticks.
This work evaluated the effect of brine concentration (3% and 9% for 90 min) combined with HPP treatments on the quality of milkfish sticks. After treating with different pressures, the APC, PBC and TVBN levels of the brine salting milkfish sticks decreased significantly, as compared to unsalted meat. However, the samples treated with 3% brine concentration combined with 300 or 400 MPa had the lower APC, PBC and TVBN levels, and the better color and texture. This result showed the combination treatment of brine salting and HPP may improve the microbiological and chemical quality of milkfish sticks.

中文摘要 I
Abstract III
誌謝 V
目錄 VIII
表目錄 X
圖目錄 XII
壹、研究動機 1
貳、文獻整理 4
一、高壓加工技術及在食品上的應用 4
二、鹽漬加工 9
三、欄柵技術 11
四、虱目魚 12
五、魚貝類之鮮度判定指標 14
六、食品中的組織胺 18
參、高壓技術對虱目魚柳衛生品質之影響及在低溫貯存時保鮮評估 28
一、前言 28
二、材料與方法 31
(一) 實驗材料 31
(二) 實驗方法 32
三、結果與討論 41
肆、高壓技術結合鹽水漬對虱目魚柳品質之影響 69
一、前言 69
二、材料與方法 72
(一) 實驗材料 72
(二) 實驗方法 73
三、結果與討論 78
伍、結論 93
陸、參考文獻 95

王文政、馮貢國、葉龍山 (2012) 鯖魚加工食品之技術開發。食品資訊，247: 66-69。
王鐘毅 (2013) 食品高壓加工技術之應用現況。食品資訊，256: 68-71。
伍迎鵑 (2020) 高壓技術結合檸檬精油對組織胺生產菌之殺菌及在鮪魚肉之保鮮應用。國立高雄海洋科技大學水產食品科學系碩士論文，高雄。
江詠薇 (2012) 臺灣冷藏宅配溫度監測實證與其對食品安全影響之探討。國立台灣海洋科技大學食品科學系碩士學位論文，基隆。
行政院農業委員會漁業署 (2005) 中華民國94年漁業年報，臺北。
行政院農業委員會漁業署 (2008-2018) 中華民國97年-107年漁業年報，臺北。
行政院衛生福利部 (2006-2019) 中華民國臺灣地區食物中毒發生與防治年報，臺北。
行政院衛生福利部 (2013) 授食字第 1021951163 號與1021956329號公告食品微生物之檢驗方法－大腸桿菌之檢驗與大腸桿菌群之檢驗。
行政院衛生福利部 (2019) 授食字第 1071303671 號公告冷凍食品類微生物衛生標準。
行政院衛生福利部 (2018)「食品中汙染物質及毒素衛生標準」網址https://mohwlaw.mohw.gov.tw。
吳清熊、周照仁、蔡憲華 (1992) 海事專校水產化學 (下冊)，華香園出版社，臺北。
李治中、鄭昌家、李孟頎 (1992) 虱目魚之產銷及加工現況與展望，虱目魚利用研討會專刊，p.1-10，國立高雄海專，高雄。
李錦楓、 陳建元、黃卓治、陳華敏、蔡滄朝、洪沄利、古國隆、曾慶瀛、李貽琳、楊正護、王正方、林志城、陳昭雄、黃至盛 (2007) 新編食品加工 (三版) ，匯華出版社，台北市。
林嬙薇 (2016) 虱目魚柳貯藏試驗及市售虱目魚產品之組織胺相關衛生品質探討。國立高雄海洋科技大學水產食品科學系碩士論文，高雄。
林靜雯 (2008) 市售水果酒之化學品質、生物胺含量及組織胺生產菌之探討。國立高雄海洋科技大學水產食品科學系碩士論文，高雄。
邱思魁 (1995) 水產品鮮度及測定方法。漁市場檢驗人員培訓班水產技術講義，p.31-47，臺灣省水產試驗所編印，基隆。
邵廣昭 (1996) 臺灣常見魚介貝類圖說 (下)-魚類。臺灣省漁業局。臺北。
孫寶年、翁秀貞、陳樹功 (1994) 臺灣地區常見魚貝類圖說 (第二版) 行政院衛生署，p.175，臺北。
張勝雄、蔡永祥、黃登福 (1996) 鯖科魚類組織胺中毒及其防止方法。漁業推廣月刊，112: 57-60
許婷儀 (2012) 不同加工方法對鹽水漬鯖魚片之衛生品質影響。國立高雄海洋科技大學水產食品科學系碩士論文，高雄。
郭鴻均 (1996) 低水分活性食品之指標微生物。海大漁推，21: 233 –242。
施明智、施耀翔、林慧生 (2011) 欄柵技術應用於豆乾加工之研究，臺灣農業化學與食品科學，49 (3): 152-158。
陳韶嵐 (2019) 高壓技術對水產品保鮮與提高食用安全性之應用。國立高雄科技大學水產食品科學系碩士論文，高雄。
曾湘媚 (2019) 高壓技術結合百里香精油對組織胺生產菌之殺菌作用與對旗魚肉的保鮮應用。國立高雄科技大學水產食品科學系碩士論文，高雄。
蔡永祥 (2010) 高壓加工技術在水產品之應用。教育部產學合作資訊網，第100501期。
蔡永祥、李憶甄 (2016) 吃龍蝦免撥殼!? 新穎食品加工-高壓加工技術，科技部科技大觀園網站https://scitechvista.nat.gov.tw/c/sZll.htm。
謝靜瑜 (2018) 高壓技術對組織胺生產菌之殺菌作用與對鮪魚肉的保鮮應用。國立高雄科技大學水產食品科學系碩士論文，高雄。
蘇茂森 (2007) 虱目魚160 (水產試驗所特刊第9號)。行政院農業委員會水產試驗所，基隆。
 |
 | Haiyuan Huang
 | Han Wu
 | Ludan Tu
 | Wenzheng Shi
 | Yinghong Qu
 | Zhihe Wang
1
1
1,2
1,2
1,2
1,2
1,2
1,2
1,2
1∗
and Ragnar L Olsen
Arnold, S. H. and Brown, W. (1978) Histamine toxicity from fish products. Advances in Food Research, 24:113-154.
Balamurugan, S., Ahmeda, R., Chibeu, A., Gao, A., Koutchma, T. and Strange, P. (2016) Effect of salt types and concentrations on the high-pressure inactivation of Listeria monocytogenes in ground chicken. International Journal of Food Microbiology, 218: 51-56.
Balasubramaniam, V. M., Farkas, D. and Turek, E. J. (2008) Preserving foods through high-pressure processing. Food Technology, 62 (11): 32-38.
Bartholomew, B. A., Berry, P. R., Rodhouse, J. C., Gilbert, R. J. and Murray, C. K. (1987) Scombrotoxic Fish Poisoning in Britain: Features of over 250 suspected incidents from 1976 to 1986. Epidemiology Infection, 99: 775-782.
Bjørn Gundersen
Calik, H., Morrissey, M.T., Reno, P. W. and An, H. (2002) Effect of high-pressure processing on Vibrio parahaemolyticus strains in pure culture and pacific oysters. Journal of Food Science, 67: 1506-1510.
Chaijan, M. (2011) Physicochemical changes of tilapia (Oreochromis niloticus) muscle during salting. Food Chemistry, 129 (3): 1201-1210.
Chen, L. C. (1990) Aquecuture in Taiwan. London: Fishing News, 119-137.
Cobb, B. F., Alauiz , I. A. and Thompson , C. A. (1973) Biochemical and microbial studies on shrimp: nitrogen and amine nitrogen analysis. Journal of Food Science, 38: 431-435.
Cruz-Romero, M., Smiddy, M., Hill, C., Kerry, J. P., and Kelly, A. L. (2004). Effects of high pressure treatment on physicochemical characteristics of fresh oysters (Crassostrea gigas). Innovative Food Science & Emerging Technologies, 5 (2): 161-169.
Daryaei, H. and Balasubramanian, V. M. (2012) Microbialdecontamination of food by high pressure processing. p.370-406. Demirci, A. and Ngadi, M. (Eds.), Microbial decontamination in the food industry, Woodhead Publishing Limited, UK.
Eerola, S., Hinkkanen, R., Lindfors, E. and Hirvi, T. (1993) Liquid chromatographic determination of biogenic amines in dry sausages. Journal of AOAC International, 76: 575-577.
Fan, H., Luo, Y., Yin, X., Bao, Y. and Feng, L. (2014) Biogenic amine and quality changes in lightly salt- and sugar-salted black carp (Mylopharyngodon piceus) fillets stored at 4°C. Food Chemistry, 159: 20-28.
FAO (2018) The state of world fisheries and aquaculture 2018 - meeting the sustainable development goals. http://www.fao.org/3/i9540en/ i9540en.pdf
FAO and WHO (2013) Public health risks of histamine and other biogenic amines from fish and fishery products. http://www.fao.org/fileadmin/ user_upload/agns/pdf/Histamine/Histamine_AdHocfinal.pdf
Flack, J. M., Peters, R., Shafi, T., Alrefai, H., Nasser, S. A. and Crook, E. (2003) Prevention of hypertension and its complications: Theoretical basis and guidelines for treatment. Journal of the American Society of Nephrology, 14 (suppl 2) S92–S98.
Gill, T. A. (1990) Objective analysis of seafood quality. Food Reviews International, 6: 681-714.
Gouygou, J. P., Sinquin, C., Etienne, M., Landrein, A. and Durand, P. (1992) Quantitative and qualitative determination of biogenic amines in fish. p. 178-186. Chap. 15 In: Burt J. R. (eds) Pelagic Fish- The Resource and its Exploitation. Fishing News. Oxford, U.K.
Gram, L. and Huss, H. H. (1996) Microbiological spoilage of fish and fish products. International Journal of Food Microbiology, 33: 121-137.
Harada, K. (1991). Fish colour. Australian Fisheries, 50: 18-19.
Hayman, M., Baxter, I., Oriordan, P. J. and Stewart, C. M. (2004) Effects of high-pressure processing on the safety, quality, and shelf life of ready-to-eat meats. Journal of Food Protection, 67 (8): 1709-1718.
He, H., Adams, R. M., Farkas, D. F. and Morrissey, M. T. (2002) Use of high-pressure processing for oyster shucking and shelf-life extension. Journal of Food Science, 67: 640-645.
Huang, H. W., Hsu, C. P., Yang, B. B. and Wang C. Y. (2013) Advances in the extraction of natural ingredients by high pressure extraction technology. Trends in Food Science & Technology, 33: 54-62
Hui, J. Y. and Taylor S. L. (1985) Inhibition of in vivo histamine metabolism in rats by foodborne and pharmacologic inhibitors of diamine oxidase, histamine N-methyltransferase, and monoamine oxidase. Toxicology and Applied Pharmacology, 81 (2): 241-249.
Hultin, H. O. (1985) Characteristics of muscle tissue. In : O. R. Fennema (Ed.). Food Chemistry, 2nd ed. p.723-789. Marcel Dekker, Inc., Madison Ave., New York.
Hwang, D. F. Chen, T. Y. Chang, S. H. Chou, S. S. Deng, J. F. and Chai, T. (1999) Biogenic amines and bacterial isolates of marlin implicated in food poisoning. Journal of the Science of Food and Agriculture, 1: 223-228.
Hwang, D. F., Chang, S. H., Shiau, C. Y. and Cheng, C. C. (1995) Biogenic amines in the flesh of sailfish (Istiophorus platypterus) responsible for scombroid poisoning. Journal of Food Science, 60:926-928.
Kim, D. H., Woo, K. B. and Ahn, D. H. (2013) Inhibitory effects of high-hydrostatic-pressure treatments on histamine production in mackerel (Scomber japonicus) muscle inoculated with Morganella morganii and Photobacterium phosphoreum. Food Control, 34 (2): 307-311.
Knorr, D. (1993) Effects of high-hydrostatic-pressure processes on food safety and quality. Food Technology, 47 (6): 156–161.
Konosu, S., Watanabe, K. and Shimizu, T. (1974) Distribution of nigrogenous constituents in the muscle extracts of eight species of fish. Nippon Suisan Gakkaishi, 40, 909-915.
Kriaek, M., Matejkova, K., Dadakova, E., Spicka, J., Vacha, F., and Vrchotova, N. (2015) Changes in the content of biogenic amines and fatty acids in high pressure–processed carp flesh (Cyprinus carpio). Journal of Food Protection, 78 (8): 1592-1596.
Kristin Lauritzsen,
Kruk, Z. A., Yun, H., Rutley, D. L., Lee, E. J., Kim, Y. J. and Jo, C. (2011) The effect of high pressure on microbial population, meat quality and sensory characteristics of chicken breast fillet. Food Control, 22: 6-12.
Lauritzsen, K., Akse, L., Gundersen, B. and Olsen, R. L. (2004) Effects of calcium, magnesium and pH during salt curing of cod (Gadus morhua L). Journal of the Science of Food and Agriculture, 84: 683–692.
Ledward, D. A. (1998). High-pressure processing of meat and fish. In K. Autio (Ed.), Fresh novel foods by high-pressure. Espoo: VVT Biotechnology and Food Research, 165-175.
Lee, Y. C., Hsieh, C. Y., Chen, M. L., Wang, C. Y., Lin, C. S and Tsai, Y. H. (2019) High pressure inactivation of histamine-forming bacteria Morganella morganii and Photobacterium phosphoreum. Journal of Food Protection, 83 (4): 621-624.
Lee, Y. C., Tsai, Y. H., Chen, S. L., Kung, H. F., Arakawa, O. and Wei, C. I. (2020) Inactivation and damage of histamine-forming bacteria by treatment with high hydrostatic pressure. Foods, 9 (3): 266.
Lee, Y. C., Kung, H. F., Wu, C. H., Hsu, H. M., Chen, H. C., Huang, T. H. and Tsai, Y. H. (2016) Determination of histamine in milkfish stick implicated in food-borne poisoning. Journal of Food and Drug Analysis, 24 (1): 63-71.
Leif Akse,
Leistner, L. (1985) Hurdle technology applied to meat products of the shelf stable product and intermediate moisture food types. Simatos, D. and Multon, J. L. (Eds.), Properties of water in foods: In relation to quality and stability, Springer Netherlands, Dordrecht, 309-329.
Leistner, L. and Gorris, G. M. (1995) Food preservation by hurdle technology. Trends in Food Science and Technology, 6: 41-46.
Matějková, K., Křížek, M., Vácha, F. and Dadáková, E. (2013) Effect of high-pressure treatment on biogenic amines formation in vacuum-packed trout flesh (Oncorhynchus mykiss). Food Chemistry, 137: 31-36.
Medina-Meza, I. G., Barnaba, C. and Barbosa-Cánovas, G. V. (2014) Effects of high pressure processing on lipid oxidation: A review. Innovative Food Science & Emerging Technologies, 22: 1-10.
Montiel, R., De Alba, M., Bravo, D., Gaya, P. and Medina, M. (2012) Effect of high pressure treatments on smoked cod quality during refrigerated storage. Food Control, 23 (2): 429-436.
Morrow, J. D., Margolies, G. R., Rowland, J. and Robert, L. J. (1991) Evidence that histamine is the causative toxin of scombroid-fish poisoning. New England J. Medicine, 324: 716-720.
Pivarnik, L. F., Thiam, M. and Ellis, P. C. (1998) Rapid determination of volatile bases in fish by using an ammonia ion-selective electrode. Journal of AOAC International, 81 (5): 1011-1022.
Post, L. S., Lee, D. A., Solberg, M., Furgang, D., Speecchio, J. and Graham, C. (1985) Development of botulinal toxin and sensory deterioration during storage of vacuum and modified atmosphere packaged fish fillets. Journal of Food Science, 50: 920-996.
Rastogi, N., Raghavarao, K. S. M. S., Balasubramaniam, V. M., Niranjan, K. and Knorr D. (2007) Opportunities and challenges in high pressure processing of foods. Critical Reviews in Food Science and Nutrition, 47: 69-112.
Rawles, D. D., Flick, G. J. and Martin, R. E. (1996) Biogenic amines in fish and shellfish. Advances in Food and Nutrition Research, 39: 329-365.
Remenant, B., Jaffres, E., Dousset, X., Pilet, M. F. and Zagorec, M. (2015) Bacterial spoilers of food: behavior, fitness and functional properties. Food Microbiology, 45: 45-53.
Saiz, A. H., Mingo, S.T., Balda, F. P. and Samson, C. T. (2008) Advances in design for successful commercial high pressure food processing. Food Australia, 60: 154-156.
Sasagawa, A., Naiki, Y., Nagashima, S., Yamakura, M., Yamazaki, A. and Yamada, A. (2006) Process for producing brown rice with increased accumulation of GABA using high-pressure treatment and properties of GABA-increased brown rice. Journal of Applied lycoscience, 53 (1): 27-33.
Shiau, C. Y., Pong, Y. J., Chiou, T. K., and Chai, T. (1997) Effect of growth on the levels of free histidine and amino acids in white muscle of milkfish (Chanos chanos). Journal of Agricultural and Food Chemistry, 45:2103-2106.
Simonin, H., Duranton, F. and de Lamballerie, M. (2012) New insights into the high-pressure processing of meat and meat products. Comprehensive Reviews in Food Science and Food Safety, 11: 285-306.
Siyuan Shen
Taylor, S. L. (1986) Histamine food poisoning: Toxicology and clinical aspects. Critical Reviews in Toxicology, 17: 91-128.
Taylor, S. L., Guthertz, L. S., Leatherwood, M. and Liber E. R. (1979) Histamine production by Klebsiella pneumoniae and an incident of scombroid fish poisoning. Applied and Environmental Microbiology, 37: 274-278.
Taylor, S. L., Stratton, J. E. and Nordlee, J. A. (1989) Histamine poisoning (Scombroid Fish Poisoning): An allergy-like intoxication. Journal of Toxicology: Clinical Toxicology, 27: 225-240.
Teixeira, B., Fidalgo, L., Mendes, R., Costa, G., Cordeiro, C., Marques, A., Saraiva, J. A. and Nunes, M. L. (2014) Effect of high pressure processing in the quality of sea bass (Dicentrarchus labrax) fillets. Pressurization Rate, Pressure Level and Holding Time. Innovative Food Science & Emerging Technologies, 22: 31-39.
Thorarinsdottir, K. A., Arason, S., Sigurgisladottir, S., Gunnlaugsson, V. N., Johannsdottir, J. and Tornberg, E. (2011) The effects of salt-curing and salting procedures on the microstructure of cod (Gadus morhua) muscle. Food Chemistery, 126 (1): 109-115.
Tsai, Y. H., Kung, H. F., Chen, H. C., Chang, S. C. and Wei, C. I. (2007) Determination of histamine and histamine-forming bacteria in dried milkfish (Chanos chanos) implicated in a food-borne poisoning. Food Chemistry, 105 (3): 1289-1296.
Tsai, Y. H., Kung, H. F., Lee, T. M., Chen, H. C., Chou, S. S. Wei, C. I. and Hwang, D. F. (2005) Determination of histamine in canned mackerel implicated in a food borne poisoning. Food Control, 16: 579-585.
Tsai, Y. H., Kung, H. F., Lee, T. M., Lin, G. T. and Hwang, D. F. (2004) Histamine-related hygienic qualities and bacteria found in popular commercial scombroid fish fillets in Taiwan. Journal of Food Protection, 67: 407-412.
USFDA (United States Food and Drug Administration) (2019) Fish and fishery products hazards and controls guidance, 4th ed. Department of Health and Human Services, Public Health Service, U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition Office of Food Safety, Washington, DC.
Wang, C. Y., Huang, H. W., Hsu, C. P. and Yang, B. H. (2016) Recent advances in food processing using high hydrostatic pressure technology. Critical Reviews in Food Science and Nutrition, 56 (4): 527-540.
Watabe, S., Kamal, M. and Hashimoto, K. (1991) Postmortem changes in ATP, creatine phosphate, and lactate in sardine muscle. Journal of Food Science, 56: 151-153&171.
Watabe, S., Ushio, H., Iwamoto, M., Yamanaka, H. and Hashimoto, K. (1989) Temperature‐Dependency of Rigor‐Mortis of Fish Muscle: Myofibrillar Mg2+‐ATPase Activity and Ca2+ Uptake by Sarcoplasmic Reticulum. Journal of Food Science, 54: 1107-1115.
Wenxian Yang
Yagiz, Y., Kristinsson, H. G., Balaban, M. O. and Marshall, M. R. (2007) Effect of high pressure treatment on the quality of rainbow trout (Oncorhynchus mykiss) and mahi mahi (Coryphaena hippurus). Journal of Food Science, 72: 509-515.
Yang, W., Shi, W., Qu, Y., Wang, Z., Shen, S., Tu, L., Huang, H. and Wu, H. (2020) Research on the quality changes of grass carp during brine salting. Food Science & Nutrition, 8 (6): 2968-2983.
Zhang, Y., Li, Q., Li, D., Liu, X. and Luo, Y. (2015) Changes in the microbial communities of air-packaged and vacuum-packaged common carp (Cyprinus carpio) stored at 4°C. Food Microbiology, 52: 197-204.