Learning Material 1807307 VIVAT KEAWDOUNGLEK

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 8: Environmental Design and Management for Food Sanitation and Safety Figure 8.9 Schematic diagram of plastic tubular digester Source: Ferrer et al., 2011. Wisakha Phoochinda, 2012, p. 72. Conclusion In the environmental sanitation design for food sanitation and safety, you should concern in these topic as follows; 1) separate raw from ready-to-eat food; 2) must be cleanable; 3) made of compatible material; 4) smooth and accessible surface; 5) must be self-draining; 6) framework not penetrated; and 7) proper ventilation. Moreover, the grease traps system should be provided to remove oil and grease from wastewater of their kitchen. For the waste of food preparation and cooking, they should be collected and disposed in the sanitary principle. In the part of the rodent and vector control in the kitchen, the integrated-pest management system should be Page | 93

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 8: Environmental Design and Management for Food Sanitation and Safety concerned including 1) identify the pest problem; 2) decide how much pest control is necessary; 3) choose an effective option; and 4) evaluate the results. Questions 1. From this picture, why it is a wrong design for food sanitation and safety and how to improve the design of this situation? 2. How to separate and dispose the waste from your kitchen? 3. If you found flies in your kitchen, how to reduce or prevent this problem in your kitchen? 4. Please describe the process of IPM for pest management in your kitchen. References Drainnet. (n.d.). Big Dipper Automatic Grease Traps. Retrieved on March 18, 2015 from http://drain-net.com/ Huffman, R. (2005) . Minimum requirement for effective food safety interventions to reduce Listeria monicytogenes contamination of ready –to- eat food. Retrieved on March 13, 2015 form http://www.card.iastate.edu /food_safety /workshop3/presentations/17_listeria_monocytogenes.pdf Ideal mat. (n.d.). Honeycomb Grease Resistant - Anti Slip Kitchen Swarf Mat. Retrieved on March 18, 2015 from http://www.idealmats.co.uk/swarf-mat- anti-slip-honeycomb-grease-resistant Irrigation training. (n.d.). An introduction to cross connection control. Retrieved on March 18, 2015 from http://www.irrigationtraining.com/introtobackflow.html Kajiwara, Co.Ltd. (n.d.). Steam Jacketed Kettle. Retrieved on March 18, 2015 from http://www.kajiwara.co.jp/en/product/etc/food.html Page | 94

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 8: Environmental Design and Management for Food Sanitation and Safety McSwane, D., Rue, N.R., & Linton, R. (2005). Essential of food safety and sanitation (4th ed.). New jersey: Pearson Education. Moerman, F. (n.d.). Hygienic design of food processing equipment and hygienic practices during maintenance operations. Retrieved on March 18, 2015 from http://www.xn--b1agapcsgv.xn--p1ai/tl_files/trakonru_content/files/ hygienic_program/Hygienic%20design%20open%20&%20closed%20equipm ent%20+%20maintenance.pdf Virginia’s Community Colleges. (n.d.). Lesson 10: Cross-Connection Control. Retrieved on March 18, 2015 from http://water.me.vccs.edu/courses/ ENV195/Lesson10_print.htm Page | 95

Learning Material 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring System for Food Sanitation and Safety At the end of the session, students must be able to 1. Describe the process of food sanitation monitoring system 2. Apply the principle of surveillance and monitoring system for food sanitation and safety in the real situation In the part 1-8 of the food sanitation and safety, several of processes and practices, for examples, the food hazards prevention, good personal hygiene, design for food sanitation and safety, and waste management in the kitchen are demonstrated to encourage the low risk of food hazard affecting to the health. However, the surveillance and monitoring system is required to confirm the practices associated with the principle of food sanitation and safety. Hence, the process of food sanitation and safety surveillance and monitoring system should be provided in this part. This information is not only increase the knowledge related to the surveillance and monitoring system in food sanitation and safety, but it is also increase the body of knowledge associated with the carrier path of environmental health, especially, the environmental health officer in the local government. 9.1 Codex Standard Generally, there are 339 standards and public measurement associated with food sanitation and safety in “Codex” body including standard, code of practice, guideline, miscellaneous blueprint (Misc), and minimal risk levels for hazardous substances (MRLs)(Codex Alimentarius Commission, 2015). For more information, please click http://www.codexalimentarius.org/ standards/en/ 9.2 Food Sanitation Monitoring System in China According to Zhou & Jin (2013), the food sanitation monitoring system and several measurements are established by the Chinese government to eliminate the food safety problem and improve the quality of food in China. The food safety standard of China shall contain (Zhou & Jin, 2013, p. 183):

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety 1) Provision on limits of pathogenic microorganisms, pesticide residues, veterinary medicine residues, heavy metals, pollutants and other substances hazardous to human health in food and food-related product; 2) Varieties, extent of use and dosage of food additive; 3) Nutrient content requirements for staple and supplementary food exclusively for infants and other particular groups of people; 4) Requirements for labels, marks and instructions relating to food safety or nutrition; 5) Hygienic requirements for food production or business operation process; 6) Quality requirements relating to food safety; 7) Methods and procedures for food inspection; and 8) Other contents which are necessary to be formulated as food safety standard. 9.3 Food sanitation and safety Surveillance in Thailand In Thailand, the process for food sanitation monitoring system for general area is described as lists follow (สำนักสขุ ำภบิ ำลอำหำรและนำ้ กรมอนำมยั , 2556 ก, หน้ำ 17- 18): 1) Initial review the situation of food sanitation and safety including the number of food establishment, environmental factor, health care data, and food laws and legislation to create the objective and the action plan for food sanitation and safety surveillance; 2) Establish the action plan for food sanitation and safety and coordinate the stakeholder including the local administrative, local and Provincial Public Health Office; 3) Survey the physical and environmental condition using the food sanitation and safety checklist; 4) Collect the sampling of food, drinking water, ice, and food contact area such as food worker’s hand and utensil for biological contamination analysis (as can be seen in appendix 2); 5) Analyze the data using the mean and percentage of food establishment which obtain the suitable level for food sanitation and safety standard; 6) Conclude the finding data and synthesize the food sanitation measurement for solving and preventing the unsuitable practice of food sanitation and safety based on the collective data; 7) Publish the finding data and approach to the target area and stakeholder for preventing the food sanitation and safety concern in brighter future. 8) Revise all of data to develop the food sanitation and safety data base in the area. For the specific activity area such as the exhibition, sport day, the process for food sanitation and safety surveillance is same as the general area. Additionally, we can be concerned with the chemical contamination in food establishment. Furthermore, the rectal swab collecting for biological analysis in food employee’s excreta of is required to control the rapidly spread of pathogen contaminated in food. Figure 9.1 provides the general process of food sanitation and safety surveillance in Thailand Page | 97

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Initial review the situation of food -Number of food sanitation and safety establishment. - Environmental factor -Health care data -Food laws and legislation Action plan establishment -List of activities for food sanitation and safety -Budged/ subsidy -Target area -Human Resource -Supporting factor Activities related to the action plan for -Physical Checklist food sanitation and safety -Biological Sampling -Chemical Testing Data Conclusion (Measurement) Data Publication Data Base (Revising and developing data for the future) Figure 9.1 General process of food sanitation and safety surveillance in Thailand Source: สำนักสขุ ำภบิ ำลอำหำรและนำ้ กรมอนำมยั , 2556 ก. Page | 98

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety 9.4 Food sanitation and safety monitoring program in Thailand In Thailand, There are two monitoring program to guarantee the food sanitation and safety including: 1) Clean Food Good Taste Clean food good taste is the usually monitoring program to guarantee the food sanitation and safety. There are fifteen requirements for food restaurant, twelve requirement for food shall, and thirty requirements for cafeteria. Additionally, the coliform bacteria must not be contaminated in food sampling and food contact surface material including food worker’s hand up to 90 percent of samplings. Figure 9.2 Clean Food Good Taste Source: สำนักสขุ ำภิบำลอำหำรและนำ้ กรมอนำมัย, 2556ข. 2) Healthy Market Healthy Market is defined as a symbols that guarantee the clean, hygiene and good sanitation in the market where is store and purchase several raw materials for food establishment, for example, poultry, pork, red meat product, fish, vegetable and fruit. This program includes the forty requirements for environmental health. Moreover, food sampling in the market must not be contaminated with the chemical hazard including formalin, salicylic acid, bleach, borax, and pesticide (or detect in security level for human health). In addition, consumer protection such as the information board for consumer health, weighing-machine calibration must be required to achieve this guarantee. Page | 99

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Figure 9.3 Healthy Market Source: สำนักสขุ ำภิบำลอำหำรและนำ้ กรมอนำมัย, 2553. Page | 100

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Conclusion To sum up, there are two types for food sanitation and safety surveillance and monitoring system. One is the surveillance and monitoring system according to the international standard including the codex standard. If the food or raw food may export in another countries, the codex standard should be concerned for the surveillance and monitoring system. The latter is the national surveillance and monitoring system for food sanitation and safety. Nowadays, the main of two criteria had been promoted to support the surveillance and monitoring of food sanitation and safety which are healthy market and clean food good taste. Healthy market is the standard for fresh market or supermarket which have several shops for fresh raw material selling. This criteria focus on the environmental sanitation of the market, providing the consumer protection information, and free of chemical contamination including formalin, salicylic acid, bleach, borax, and pesticide (or detect in security level for human health). Meanwhile, the clean food good taste is the standard applied in the food restaurant, canteen, or food shop related with the processing for the ready- to-eat food. It is therefore to see that the latter criteria focus on the biological contamination and the good of personal hygiene. Questions; 1. Please explain the process of surveillance for the food sanitation and safety. 2. If you create the fresh market, how to build your market according to the relevant standards. 3. If you export the raw food to the USA, how to monitor your product which is not the effect to the health? 4. Please define the main criteria of clean food good taste. 5. How to apply the surveillance and monitoring system if you are an owner of the food restaurant? Page | 101

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety References Codex Alimentarius Commission. (2015). Codex Standard. Retrieved on April 7, 2015 from http://www.codexalimentarius.org/standards/en/ Zhou, J. & Jin, S. (2013). Food safety management in China. China: Zhejiang University Press. สำนักสขุ ำภิบำลอำหำรและนำ้ กรมอนำมัย. (2553). ค่มู ือตลาดสดน่าซ้ือ. กรุงเทพมหำนคร: โรงพิมพ์ชุมนุมสหกรณก์ ำรเกษตรแห่งประเทศไทย. สำนักสขุ ำภบิ ำลอำหำรและนำ้ กรมอนำมยั . (2556 ก). ค่มู ือการปฏิบตั ิงานดา้ นสขุ าภิบาลอาหาร และนา้ สาหรบั สาธารณสขุ อาเภอ: เลม่ ที่ 2 แนวทางการเผา้ ระวงั สขุ าภิบาลอาหาร. กรุงเทพมหำนคร: โรงพิมพ์ชุมนุมสหกรณก์ ำรเกษตรแห่งประเทศไทย. สำนักสขุ ำภบิ ำลอำหำรและนำ้ กรมอนำมัย. (2556 ข). ค่มู ืออาหารสะอาด รสชาติอร่อย. กรุงเทพมหำนคร: โรงพิมพ์ชุมนุมสหกรณ์กำรเกษตรแห่งประเทศไทย. Suggested Website http://foodsan.anamai.moph.go.th/main.php?filename=media2 Page | 102

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Appendix 1 เอกสารแนบทา้ ยประกาศกรมวทิ ยาศาสตรก์ ารแพทย์ เรือ่ ง เกณฑค์ ณุ ภาพทางจลุ ชีววิทยาของอาหารและภาชนะสัมผัสอาหาร ฉบับที่ 2 1. อำหำรดบิ หมำยถงึ อำหำรท่ยี งั บริโภคไม่ได้ ต้องผ่ำนกำรทำให้สกุ หรือกำรเตรียมด้วยกรรมวิธี ใดๆ ก่อนบริโภค 1.1 เน่ือสดของสตั ว์/สตั ว์ปี ก/ เช่น เน้ือหมู เน้ือไก่ เคร่ืองใน แช่เยน็ หรือแช่แขง็ จำนวนจุลินทรีย์/กรัม น้อยกว่ำ 5X106 MPN Escherichia coli / กรัม น้อยกว่ำ 100 Staphylococcus aureus / กรัม น้อยกว่ำ 100 Clostridium perfringens / กรัม น้อยกว่ำ 1,000 Salmonella spp. / 25 กรัม ไม่พบ Campylobacter (1) /25 กรัม ไม่พบ 1.2 เน้ือสดของสตั ว์นำ้ แช่เยน็ หรือแช่แขง็ จำนวนจุลินทรีย์ / กรัม น้อยกว่ำ 1 x 106 MPN Escherichia coli / กรัม น้อยกว่ำ 3 Staphylococcus aureus / กรัม น้อยกว่ำ 100 Salmonella spp. /25 กรัม ไม่พบ Vibrio cholerae /25 กรัม ไม่พบ Vibrio parahaemolyticus (2) /25 กรัม ไม่พบ Listeria monacytogenes /25 กรัม ไม่พบ 1.3 เน้ือสตั ว์ชนิดต่ำงๆ ท่ผี ่ำนกระบวนกำรทำให้แห้ง ปริมำณนำ้ อสิ ระในอำหำร (aw) น้อยกว่ำ 0.86 จำนวนยสี ต์และรำ /กรัม น้อยกว่ำ 100 MPN E. coli /กรัม น้อยกว่ำ 3 Staphylococcus aureus /กรัม น้อยกว่ำ 100 Salmonella spp. /25 กรัม ไม่พบ 1.4 ไข่สด Salmonella spp. / 25 กรัม ไม่พบ Page | 103

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety 1.5 อำหำรพร้อมปรุงหรืออำหำรอ่นื ๆ ท่มี ีอำหำรดิบเป็นส่วนประกอบหรือส่วนผสม จำนวนจุลินทรีย์ / กรัม น้อยกว่ำ 1 x 106 MPN Escherichia coli / กรัม น้อยกว่ำ 100 Staphylococcus aureus / กรัม น้อยกว่ำ 100 Clostridium perfringens / กรัม น้อยกว่ำ 1,000 Bacillus cereus / กรัม น้อยกว่ำ 1,000 Salmonella spp. / 25 กรัม ไม่พบ Vibrio cholerae / 25 กรัม ไม่พบ Vibrio parahaemolyticus (2) /25 กรัม ไม่พบ 2. อำหำรพร้อมบริโภค น้อยกว่ำ 1 x 106 2.1 อำหำรดบิ ท่เี ตรียมหรือปรุงในสภำพบริโภคได้ทนั ที น้อยกว่ำ 500 2.1.1 ผัก ผลไม้ สลัด ส้มตำ น้อยกว่ำ 1 x 104 จำนวนจุลินทรีย์ / กรัม น้อยกว่ำ 100 จำนวนรำ / กรัม น้อยกว่ำ 100 จำนวนยสี ต์ (3) / กรัม ไม่พบ MPN Escherichia coli / กรัม ไม่พบ Staphylococcus aureus / กรัม Salmonella spp. /25 กรัม Listeria monacytogenes (4) /25 กรัม 2.1.2 อำหำรทะเล เช่น ปลำ กุ้ง ปลำหมึก หอย ซำชิมิ จำนวนจุลินทรีย์ / กรัม น้อยกว่ำ 1 x 105 MPN Escherichia coli / กรัม น้อยกว่ำ 3 Staphylococcus aureus / กรัม น้อยกว่ำ 100 Salmonella spp. /25 กรัม ไม่พบ Vibrio cholerae / 25 กรัม ไม่พบ Vibrio parahaemolyticus / 25 กรัม ไม่พบ Listeria monacytogenes / 25 กรัม ไม่พบ Page | 104

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety 2.2 อำหำรปรุงสกุ หรืออำหำรท่ผี ่ำนกรรมวิธเี กบ็ รักษำ 2.2.1 ขนมหวำน ผกั ผลไม้ (ดอง แช่อ่มิ เช่ือม กวน แห้ง) จำนวนยสี ต์และรำ / กรัม น้อยกว่ำ 1,000 MPN Escherichia coli / กรัม น้อยกว่ำ 3 Staphylococcus aureus / กรัม น้อยกว่ำ 100 Salmonella spp. /25 กรัม ไม่พบ 2.2.2 ขนมปังมีไส้หรือไม่มีไส้ อำจผสมวัตถุอ่นื ท่ไี ม่เป็นอนั ตรำยต่อสุขภำพ เช่น ลูกพรุน ลูกเกด ชอ็ กโกแลต จำนวนจุลินทรีย์ / กรัม น้อยกว่ำ 1 x 104 จำนวนยีสต์และรำ / กรัม น้อยกว่ำ 10 MPN Escherichia coli / กรัม น้อยกว่ำ 3 Staphylococcus aureus / กรัม น้อยกว่ำ 10 Salmonella spp. / 25 กรัม ไม่พบ 2.2.3 อำหำรหมักพ้ืนเมืองท่ีเป็ นผลิตภัณฑ์จำกสัตว์ เช่น แหนม กะปิ ปลำร้ำ ปลำจ่อม ส้มฟัก บูดู MPN Escherichia coli / กรัม น้อยกว่ำ 3 Staphylococcus aureus / กรัม น้อยกว่ำ 100 Clostridium perfringens / กรัม น้อยกว่ำ 1,000 Bacillus cereus / กรัม น้อยกว่ำ 1,000 Salmonella spp. / 25 กรัม ไม่พบ 2.2.4 อำหำรปรุงสกุ ท่วั ไป เช่น อำหำรปรุงสำเรจ็ (ประเภทข้ำวแกง กว๋ ยเตี๋ยว ขนมจีน) ยำ ไส้กรอก หมูยอ ปูอดั ปลำหมึกปรุงรส ซูชิ จำนวนจุลินทรีย์ / กรัม น้อยกว่ำ 1 x 106 MPN Escherichia coli / กรัม น้อยกว่ำ 3 Staphylococcus aureus / กรัม น้อยกว่ำ 100 Clostridium perfringens / กรัม น้อยกว่ำ 100 Bacillus cereus / กรัม น้อยกว่ำ 100 Salmonella spp. / 25 กรัม ไม่พบ Vibrio cholerae / 25 กรัม ไม่พบ Page | 105

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Vibrio parahaemolyticus (2) /25 กรัม ไม่พบ Listeria monacytogenes / 25 กรัม ไม่พบ 2.2.5 อำหำรท่ผี ่ำนกระบวนกำรทำให้แห้ง อบหรือทอด เช่น อำหำรอบกรอบ อำหำรทอดกรอบ นำ้ พริก ปริมำณนำ้ อสิ ระในอำหำร (aw) น้อยกว่ำ 0.86 จำนวนยสี ต์และรำ / กรัม น้อยกว่ำ 100 MPN Escherichia coli / กรัม น้อยกว่ำ 3 Staphylococcus aureus / กรัม น้อยกว่ำ 10 Clostridium perfringens / กรัม น้อยกว่ำ 100 Bacillus cereus / กรัม น้อยกว่ำ 1,000 Salmonella spp. / 25 กรัม ไม่พบ 3. นำ้ ด่มื ท่ไี ม่ได้บรรจุในภำชนะปิ ดสนิท เช่น นำ้ ผ่ำนเคร่ืองกรอง ไม่พบ Escherichia coli / 100 มิลลิลิตร ไม่พบ Staphylococcus aureus / 100 มิลลิลิตร ไม่พบ Salmonella spp. / 100 มลิ ลิลิตร ไม่พบ Clostridium perfringens / 100 มิลลิลิตร 4. เคร่ืองด่มื ท่ไี ม่ได้บรรจุในภำชนะปิ ดสนิท เช่น เคร่ืองด่มื รถเขน็ / แผงลอยริมถนน / ร้ำน อำหำรฯ จำนวนยสี ต์ / มลิ ลิลิตร น้อยกว่ำ 5,000 จำนวนเช้ือรำ / มลิ ลิลิตร น้อยกว่ำ 100 Escherichia coli / มลิ ลิลิตร ไม่พบ Staphylococcus aureus / 0.1 มิลลิลิตร ไม่พบ Salmonella spp. / 25 มลิ ลิลิตร ไม่พบ Clostridium perfringens / มิลลิลิตร น้อยกว่ำ 100 Bacillus cereus / มิลลิลิตร น้อยกว่ำ 100 5. ภำชนะสมั ผัสอำหำร (5) น้อยกว่ำ 1,000 จำนวนจุลินทรีย์ / ช้ินภำชนะ ไม่พบ Staphylococcus aureus / ช้ินภำชนะ ไม่พบ Salmonella spp. / ช้ินภำชนะ Page | 106

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety 6. พ้ืนผวิ สมั ผสั อำหำร ไม่พบ Escherichia coli / 50 ตำรำงเซนตเิ มตร ไม่พบ Staphylococcus aureus / 50 ตำรำงเซนติเมตร ไม่พบ Salmonella spp. / 50 ตำรำงเซนตเิ มตร ไม่พบ Clostridium perfringens / 50 ตำรำงเซนติเมตร ไม่พบ Bacillus cereus / 50 ตำรำงเซนตเิ มตร ไม่พบ 7. มือผู้สัมผัสอำหำร (6) ไม่พบ Escherichia coli / มอื ไม่พบ Staphylococcus aureus / มอื Salmonella spp. / มอื หมายเหตุ (1) Compylobacter jejuni / coli (2) เฉพำะอำหำรทะเลหรืออำหำรท่มี ีอำหำรทะเลเป็นองค์ประกอบ (3) เฉพำะผลไม้หรือมีผลไม้เป็นสว่ นประกอบ (4) เฉพำะผักหรือมผี ักเป็นองค์ประกอบ (5) ส่มุ ตัวอย่ำงชนิดเดียวกนั อย่ำงน้อย 4 ช้ินภำชนะ ยกเว้น เขยี ง/มดี /ครก/ภำชนะท่มี ีขนำดใหญ่ สมุ่ ตัวอย่ำง 1 ช้ิน (6) swab มือข้ำงท่ถี นัด 1 มือ กรณที ต่ี รวจพบ Staphylococcus aureus หรือ Clostridium perfringens หรือ Bacillus cereus ต่อ กรัมหรือมิลลิลิตรน้อยกว่ำ 10 น้อยกว่ำ 100 หรือ น้อยกว่ำ 1,000 ให้หมำยถงึ ตรวจไม่พบเช้ือ เหล่ำน้ันใน 0.1, 0.01 หรือ 0.001 กรัม หรือมิลลิกรัม ตำมลำดบั Page | 107

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Appendix 2 ประเภทอาหารและการเก็บ ประเภทอาหาร การเก็บตวั อยา่ ง 1.อาหารคาว 1.1 พร้อมรับประทำน เช่น ข้ำวแกง เกบ็ ท่วี ำงจำหน่ำย 1.2 อำหำรจำนเดยี ว เช่น ข้ำวมนั ไก่ เกบ็ รวม 1 กรัม 1.3 กว๋ ยเต๋ยี ว เกบ็ ผักโรย เน้ือหมู หรือลูกช้ิน 1.4 อำหำรตำมส่งั เกบ็ ผักท่ที ำนคู่กนั 1.5 ขนมจีน เกบ็ ผกั และเส้น 1.6 โจก๊ ต้มเลือดหมู เคร่ืองใน เกบ็ เคร่ืองในหมู หรือหมูท่ลี วกไว้ 1.7 อำหำรอสี ำน เกบ็ ผักดบิ ต่ำงๆ 1.8 ขำยอำหำรหลำยอย่ำง ให้เลือกอำหำรท่เี ส่ียง 2. อาหารหวาน 2.1 ขนมนำ้ แขง็ เกบ็ พวกท่ใี ช้มือสมั ผสั มำกท่สี ดุ 2.2 ขนมกะทิ เกบ็ ตวั ขนม 2.3 ไอศกรีม เกบ็ ตวั อย่ำงไอศกรีม ผลไม้ท่โี รยหน้ำ (ถ้ำมี) 2.4 อำหำรว่ำง เกบ็ อำหำรท่พี ร้อมรับประทำน 2.5 ขนมหวำนแห้ง เกบ็ ท่คี ิดว่ำเส่ยี ง หมายเหตุ : ข้อกำหนดในกำรกำรตรวจสอบแบคทเี รียในอำหำร จะต้องใช้ตัวอย่ำง 10 ตัวอย่ำงต่อ ร้ำนค้ำหรือแผงลอยจำหน่ำยอำหำร 1 ร้ำน ประกอบด้วย อำหำร 5 ตัวอย่ำง ภำชนะ 3 ตัวอย่ำง (ตวั อย่ำงละไม่ต่ำกว่ำ 4 ช้ิน) และมือของผู้สมั ผสั อำหำร 2 ตวั อย่ำง ท้งั น้ีผลกำรตรวจสอบควำม สะอำดจะต้องผ่ำนร้อยละ 90 ของตัวอย่ำงท่ตี รวจ Page | 108

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety ตวั อยา่ งตารางการเก็บตวั อย่างอาหาร ภาชนะ และน้าด่ืม ของร้านอาหารและแผงลอย ประเภทของตวั อยา่ งอาหาร ชว่ งเวลา อาหาร น้าดืม่ น้าแข็ง นา้ หวาน มือผู้ปรุง ภาชนะ แก้วน้า สถานทเี่ กบ็ (ต.ย.) (ต.ย.) (ต.ย.) (ต.ย.) (ต.ย.) หยิบจับ (ต.ย.) ท่ีเกบ็ (ต.ย.) 1. ร้ำนอำหำร/ 2 คร้งั ต่อ 2 1 1 1 2 2 1 แผงลอย ปี (5 ช้นิ ) (5 ช้นิ ) 2. โรงอำหำร 2 คร้งั ต่อ 2 1 1 1 2 2 1 (เกบ็ ทกุ แผง) ปี (5 ช้นิ ) (5 ช้นิ ) 3. โรงคร้ว - 4 -- - 3 21 โรงพยำบำล คำว 2 (5 ช้นิ ) (5 ช้นิ ) สำยยำง 2 หวำน 1 แหล่งท่ีมา: สำนักสขุ ำภิบำลอำหำรและนำ้ กรมอนำมัย, 2556 ข, หน้ำ 31-33. Page | 109

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Appendix 3 เกณฑ์มาตรฐานตลาดสดน่าซือ ส้าหรบั อาหารสดที่จ้าหน่ายในตลาดสด จะต้องไม่พบตรวจไม่พบสำรเคมีดังต่อไปน้ี 1) สำรฟอร์มำลิน (นำ้ ยำดองศพ) 2) สำรกนั รำ (ซำลิซิลิก) 3) สำรบอแรกซ์ 4) ยำฆ่ำแมลง หรือตรวจพบในเกณฑท์ ่ปี ลอดภัย 5) ตรวจไม่พบสำรเร่งเน้ือแดงในเน้ือหมู (สง่ ตรวจทำงห้องปฏบิ ัตกิ ำร) หำกต้องมีกำรเกบ็ ตัวอย่ำงอำหำรในตลำดสด เพ่ือตรวจสอบกำรปนเป้ื อนสำรเคมี ควรจำแนกดัง ตำรำงต่อไปน้ี ดัชนกี ารปนเป้อื นทางเคมี ชนดิ / ประเภทของอาหาร ทต่ี ้องการทดสอบ บอแรกซ์ เน้ือสตั ว์ ผลิตภณั ฑท์ ่ที ำจำกเน้ือสตั ว์ และอ่นื ๆ เช่น หมูสด หมูบด ปลำบด ทอดมนั ไส้กรอก แป้ งกรุบ ทบั ทมิ กรอบ ย่ำฆ่ำแมลง ผลไม้ดอง ลูกช้ิน เป็นต้น ฟอร์มำลีน ผักสด ผลไม้สด ปลำแห้ง เป็นต้น อำหำรทะเลสด ผกั สด ผลไม้สด เน้ือสตั ว์สด เช่น สไบนำง สำรฟอกขำว (ผ้ำข้ีร้ิว) ปลำหมึกกรอบ (ปลำหมึกแช่ด่ำงหรือปลำหมกึ เยน็ ตำโฟ) เป็นต้น สำรกนั รำ ถ่ัวงอก ขิงซอย ยอดมะพร้ำว กระท้อน หน่อไม้ดอง นำ้ ตำล กรดแร่อสิ ระในนำ้ ส้มสำยชู มะพร้ำว ทุเรียนกวน เป็นต้น มะม่วงดอง ผกั ดอง ผลไม้ดอง เป็นต้น (นำ้ ส้มสำยชูปลอม) นำ้ ส้มสำยชู นำ้ ส้มพริกดอง นำ้ มะนำวเทยี ม เป็นต้น สผี สมอำหำร ไส้กรอก กุ้งแห้ง แหนม กุนเชียง ลูกช้ิน ปลำแห้ง ข้ำวเกรียบ เป็ นต้ น Page | 110

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 9: Surveillance and Monitoring Method for Food Sanitation and Safety Appendix 4 เลขสารบบอาหาร ตามระเบยี บส้านักงานคณะกรรมการอาหารและยา วา่ ดว้ ยการดา้ เนนิ การเกย่ี วกับเลขสารบบอาหาร พ.ศ. ๒๕๕๗ หมายเหตุ: สถานะของสถานทีผ่ ลิต/ นาเขา้ และหน่วยงานทีอ่ นุญาต มี 4 ประเภท ไดแ้ ก่ -หมำยเลข 1 หมำยถงึ สถำนท่ผี ลติ อำหำร ซ่งึ สำนักงำนคณะกรรมกำรอำหำรและยำ เป็นผ้อู นุญำต -หมำยเลข 2 หมำยถงึ สถำนท่ผี ลติ อำหำร ซ่งึ จงั หวดั เป็นผ้อู นุญำต -หมำยเลข 3 หมำยถงึ สถำนท่นี ำเข้ำอำหำร ซ่งึ สำนักงำนคณะกรรมกำรอำหำรและยำ เป็นผ้อู นุญำต - หมำยเลข 4 หมำยถงึ สถำนท่นี ำเข้ำอำหำร ซ่ึงจงั หวัดเป็นผู้อนุญำต หน่วยงานทีอ่ อกเลขสารบบอาหาร มี 2 ประเภท ไดแ้ ก่ -หมำยเลข 1 หมำยถงึ อำหำรท่ไี ด้รับเลขสำรบบอำหำรจำกสำนกั งำนคณะกรรมกำรอำหำรและยำ -หมำยเลข 2 หมำยถงึ อำหำรท่ไี ด้รับเลขสำรบบอำหำรจำกจงั หวดั Page | 111

Learning Material 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory At the end of the session, students must be able to 1. Describe the process of food sanitation laboratory 2. Define the suitable method for analysis of food sanitation and safety 3. Apply the food sanitation and safety laboratory the real situation Food hazard analysis is the requirement for food sanitation and safety surveillance and monitoring system according to the job descriptions of the environmental health officer in order to reduce and prevent the prevalence of foodborne disease in the community. Hence, they need to know about the principle of food sanitation and safety laboratory to apply this knowledge for finding and analyzing the cause of food hazard contamination. For this part, the first is the principle of biological analysis according to the standard of most probable number or MPN method. And the latter, we focus for the analysis of chemical contamination using the test-kits developed by the department of medical science, ministry of public health. 10.1 Introduction of Food microbiology (Cappuccino &Sherman,2008, p 311) Microbiologists have always been aware that foods, especially milk, have served as important inanimate vectors in transmission of disease. Food contain the organic nutrients that provide an excellent medium to support the growth and multiplication of microorganism under suitable temperatures. Food and dairy products may be contaminated in food in a Fariety of ways and from a variety of sources: 1) Soil and Water Food-borne organisms that may be found in soil and water and that may contaminate food are members of the genera Alcaligenes, Bacillus, Critorbactor, Clostridium. The common soil and water molds include, Rhizopus, Pennicillium, Fusarium. 2) Food utensils The type of microorganism found on utensils depends on type of food and the manner in which the utensil were handle. 3) Enteric Microorganism of human and animal

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory The major member of this group are Becteroides, Lactobacillus, Clostridium, Escherichia, Samonella, Shigella, Staphylococus. 4) Food Handlers People who handle foods are especially likely to contaminate them because microorganism on hands and clothing are easily transmitted. A major offending organism is Staphylococus, which is generally found on hand and skin, and in the upper respiratory tract. Food handlers with poor personal hygiene and unsanitary habits are most likely to contaminate foods whit enteric organism. 5) Animal hides and feed Microorganisms found in water, soil, feed, dust, and fecal debris can be found on animal hides. Infected hides may serve as a source of infection for worker, or the microorganisms may migrate into the musculature of the animal and remain variable following its slaughter. To determine the number of microorganism present in food products, the standard plate count using the several dilution is demonstrated. 10.2 Enumeration of Escherichia coli and the Coliform Bacteria (U.S. Food and Drug Administration, 2015) Escherichia coli, originally known as Bacterium coli commune, was identified in 1885 by the German pediatrician, Theodor Escherich . E. coli is widely distributed in the intestine of humans and warm-blooded animals and is the predominant facultative anaerobe in the bowel and part of the essential intestinal flora that maintains the physiology of the healthy host . E. coli is a member of the family Enterobacteriaceae , which includes many genera, including known pathogens such as Salmonella, Shigella, and Yersinia. Although most strains of E. coli are not regarded as pathogens, they can be opportunistic pathogens that cause infections in immunocompromised hosts. There are also pathogenic strains of E. coli that when ingested, causes gastrointestinal illness in healthy humans Figure 10.1 Escherichia coli Source: Dreamstime, 2000. Page | 113

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Although coliforms were easy to detect, their association with fecal contamination was questionable because some coliforms are found naturally in environmental samples. This led to the introduction of the fecal coliforms as an indicator of contamination. Fecal coliform, first defined based on the works of Eijkman is a subset of total coliforms that grows and ferments lactose at elevated incubation temperature, hence also referred to as thermotolerant coliforms. Fecal coliform analyses are done at 45.5°C for food testing, except for water, shellfish and shellfish harvest water analyses, which use 44.50C . The fecal coliform group consists mostly of E. coli but some other enterics such as Klebsiella can also ferment lactose at these temperatures and therefore, be considered as fecal coliforms. The inclusion of Klebsiella spp in the working definition of fecal coliforms diminished the correlation of this group with fecal contamination. As a result, E. colihas reemerged as an indicator, partly facilitated by the introduction of newer methods that can rapidly identify E. coli. Currently, all 3 groups are used as indicators but in different applications. Detection of coliforms is used as an indicator of sanitary quality of water or as a general indicator of sanitary condition in the food-processing environment. Fecal coliforms remain the standard indicator of choice for shellfish and shellfish harvest waters; and E. coli is used to indicate recent fecal contamination or unsanitary processing. Almost all the methods used to detect E. coli, total coliforms or fecal coliforms are enumeration methods that are based on lactose fermentation. The Most Probable Number (MPN) method is a statistical, multi-step assay consisting of presumptive, confirmed and completed phases. In the assay, serial dilutions of a sample are inoculated into broth media. Analysts score the number of gas positive (fermentation of lactose) tubes, from which the other 2 phases of the assay are performed, and then uses the combinations of positive results to consult a statistical table, to estimate the number of organisms present. 10.3 Swab Technique (กรมวิทยาศาสตร์การแพทย์, 2551, 24) Swab technique is the method for sampling the any food container, utensils that their surface will be contact with the food including the raw food and ready-to-eat food. Moreover, this method can be collect the microbiological contamination in human fingers or any surface that contact with the food directly. The process for swab technique is provided as lists follows (see in figure 12.2 to 12.3); 10.3.1 Clean and sanitize your hand when the swab sampling will be beginning. 10.3.2 Use the sterile cotton bud and dip into the sterile dilution water (e.g. peptone water or phosphate buffer). 10.3.3 Then, scotch the cotton bud into the surface as list below; -For hand, select one hand that usually use for food preparation and scotch the cotton bud in this hand; -For the food container, scotch the cotton bud in the bottom of a pocket around 2X2 square of inch; -For the cup and drinking water container, scotch the cotton bud that the width of this sampling from the topside is 1 inch including the inside and the outside of this container; -For any food utensil, scotch the cotton bud into the surface around 2X2 square of inch; Page | 114

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 10.3.4 Put the cotton bud into the sterile dilution water container, then break the tip of cotton bud; 10.3.5 Close the dilution water container. 10.4 Food and food equipment sampling A sample unit consists of a minimum of 100 g and is usually a consumer-size container of product. Take sample units at random to ensure that a sample is representative of the lot. When using sample containers, submit a control consisting of one empty sample container that has been exposed to the same conditions as those under which the sample was collected. Collect more than one sample unit from large institutional or bulk containers when the number of sample units required exceeds the number of containers in the lot. A sample unit will consist of more than one container when containers are smaller than 100 g (U.S. Food and Drug Administration, 2015) According to the clean food good taste standard in Thailand, the food and food equipment sampling is shown in table 10.1 Table 10.1 Food and food equipment sampling referred to the clean food good taste standard. Type of sampling Number of sampling Sampling technique Hand of Chief select one hand that usually use Swab for food preparation for one Food Container man Swab More than 4 pieces/ one (on the bottom of food Drinking water container dilution water/ a food shall or container around 2X2 inch) cafeteria Swab Small utensil More than 4 pieces/ one (the width of swab area from (e.g. spoon, chopsticks) dilution water/ a food shall or the topside is 1 inch including Large utensil cafeteria the inside and the outside of (e.g. cutting board) the container) Ready to eat food /desert More than 4 pieces or more Swab Ice than 4 pairs/ one dilution water/ (the area of this swab is 2x2 Diet Broth a food shall or cafeteria inch) 1 piece/ one dilution water/ a Swab food shall or cafeteria (the area of this swab is 2x2 inch) More than 100 g - More than 100 g - More than 100 ml - Source: U.S. Food and Drug Administration, 2015. Page | 115

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory For the food sampling, the risk or food contamination of the risk of cross contamination are considered for food sampling. The ready-to-eat food decorated or mixed by the raw material such as a fresh fruit or fresh vegetable is the significant sampling that can be contaminated with the microorganism. Additionally, the ready-to- eat food that provide in the room temperature more than 4 hours is a high risk for the multiply of microorganism in food. Hand Hand sample Food Containers Drinking Water Container Food container sample Drinking water container sample Page | 116

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Small Utensil Sample Small Utensil Large Utensil Large Utensil Sample Figure 10.2 Food and food equipment sampling referred to the clean food good taste standard Source: Adapted from Cappuccino & Sherman, 2008. Page | 117

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory The Area of food container for swab sampling one hand that usually use for food preparation The width of swab area from the topside is 1 inch including the inside and the outside of the Drinking water container 1 inch Page | 118

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory The Area of Swab technique for small utensil (2x2 inch) The Area of Swab technique for large utensil (2x2 inch) 2x2 inch Figure 10.3 Swab sampling area for food sanitation laboratory Source: Adapted from Cappuccino & Sherman, 2008. 10.5 The Most Probably Number Test (MPN) (Cappuccino &Sherman, 2008) The three basic to detect coliform bacteria in food are the presumptive, confirmed, and completed. The test are performed sequentially on each sample under analysis. They detect the presence of coliform bacteria (indicators of fecal contamination), the gram-negative. Non-spore-forming bacilli that ferment lactose with the production of acid and gas that is detectable following a 24-hour incubation period at 37 0c. 10.5.1 Presumptive test The presumptive test is specific for detection of the coliform bacteria. Measured aliquots of water and food to be tested are added to a lactose fermentation broth containing an inverted gas vial. Because these bacteria are capable of using lactose as a carbon source (the other enteric organism are this medium. In this experiment, you Page | 119

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory will use lactose fermentation broth containing an inverted Durham gas for gas collection) Tubes of this lactose medium are incubated with 10-ml, 1-ml, 0.1-ml aliquots of water and food sampling. The series consists of at least three group, each composed for five tubes of the specified medium. The tube in each group are then inoculated with designated volume of the water or food sample. The greater the number of tubes per group, the greater the sensitivity of the test. Development of gas in any of the tubes is presumptive evidence of the present of coliform bacteria in the sample. The presumptive test also enables the microbiologist to obtain some idea of the number of the most probable number (MPN) test. The number of tubes in each group that show gas following the incubation periods (See in table 12.2). Positive Negative Figure 10.4 The positive and negative of Durham gas collection tube in the Lactose Broth for MPN Method Source: Spilatro, 2004. 10.5.2 The confirmed test The presence of a positive or doubtful presumptive test immediately suggests that the sample is non-potable. Confirmation of these results is necessary because positive presumptive tests may be the result if organism of non-coliform origin that are not recognized as indicators of the fecal pollution. Page | 120

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory The confirmed test required that selective and differential media such as the gas fermentation in the specific media such as brilliant green lactose bile broth (BGLB) from the positive lactose broth is the alternative method to confirm the total coliform bacteria that can be contaminated in any food. Positive Negative Figure 10.5 The positive and negative of Durham gas collection tube in the brilliant green lactose bile broth (BGLB) for MPN Method Source: Soleris, 2018. Additionally, an isolated colony is picked up from the confirmatory test and streak on the gram negative media such as EC-Medium Broth that the fecal coliform bacteria can be produced a gas in this broth. Page | 121

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Positive Negative Figure 10.6 The positive of Durham gas collection tube in the EC-medium for MPN Method 10.5.3 The complete test The completed test is the final analysis of the water and food sample. The Eosin- methylene blue (EMB agar) or Endo agar be streaked form the positive of the confirmed test. EMB form a complex that precipitates out onto the coliform bacteria such as E. coli, producing dark center and a green metallic sheen, while the Endo agar provided a dark pink colonies in this media. Additionally, the gram staining is used for analyzing the microorganism in the completed test. Figure 10.7 Metallic green sheen colonies of the E. coli specified in the EMB agar Source: Microbe Online, 2019. Page | 122

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 10.6 Food Sanitation and Safety Laboratory: MPN method 10.6.1 Sample and Culture Lab I (presumptive) Lab II (confirmed) Lab III (completed) -Food Sample: 1 sample 24-hours-old of positive 24-hours-old of positive lactose broth culture form EC-medium culture -Food container: 5 pieces each of the three series form each of the three per a dilution water per form the presumptive test series form the group presumptive test -Utensils: 1-5 pieces per a dilution water per group 10.6.2 Media Lab I (presumptive) Lab II (confirmed) Lab III (completed) -3 double strength lactose 12 single strength - EMB agar fermentation Broth (LB2X) Brilliant Green lactose per one of sample per group bite broth (BGLB) per one of sample per group -9 single strength lactose fermentation broth (LB1X) -12 single strength of EC- per one of sample per group medium broth per one of sample per group 10.6.3 Equipment/ Reagent Lab I (presumptive) Lab II (confirmed) Lab III (completed) -Sterile Cotton bud: 10 -Inoculation Loop -Inoculation Loop pieces per group -Glassware make pencil -Glassware make pencil -Staining tray -Sterile dilution water -Lens paper (peptone) : 5 bottoms per -Glass slide group -Microscope -Gram iodine -Bunsen burner -12 test tube per sample -Sterile 10-ml, 1-ml, and 0.1 ml pipettes. Page | 123

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Lab I (presumptive) Lab II (confirmed) Lab III (completed) - 95% ethyl alcohol -Sterile glassware and - Safranin glassware make pencil -Crystal violet -Pertri dish -Bibulous Paper -Sterile bag -Stomacher/ Blender 10.6.4 Procedure Sample preparation 1) Solid Food sampling: scotch the cotton bud into the solid food sample as same as the swab technique; 2) Liquid sample: mix the liquid food sample and then transfer the 10-ml, 1-ml, and 0.1-ml of sample directly; 3) Swab sampling: shake the dilution water by the hand and then transfer the 10-ml, 1-ml, and 0.1-ml of sample directly. Lab I: Presumptive test 1) Set up three separate series consisting of three groups, a total of 9 tubes per series per sample, in the test tube rack; for each tube, label the sampling source and volume of sample inoculated as illustrated Series 1: Hand-swab sample 3 tube of LB2X-10 ml Series 2: Food/beverage sample 3 tube of LB1X-1 ml Series 3: Utensil-swab sample 3 tube of LB1X-0.1 ml Series 4: Food container- swab sample 3 tube of LB2X-10 ml 3 tube of LB1X-1 ml 3 tube of LB1X-0.1 ml 3 tube of LB2X-10 ml 3 tube of LB1X-1 ml 3 tube of LB1X-0.1 ml 3 tube of LB2X-10 ml 3 tube of LB1X-1 ml 3 tube of LB1X-0.1 ml Page | 124

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 2) Using a 10-ml pipette, transfer 10-ml of hand-swab sample to the three tubes labeled LB2X-10ml; 3) Using a 1-ml pipette, transfer 1-ml of hand-swab sample to the three tubes labeled LB1X-1ml; 4) Using a 0.1-ml pipette, transfer 0.1-ml of hand-swab sample to the three tubes labeled LB1X-0.1ml; 5) Repeat step of 2 to 4 for the food/beverage sample, utensil-swab sample, and food container-swab sample; 6) Incubate all tubes for 24-48 hours at 370C. Lab II confirmed test Total Coliform Bacteria 1) Prepare the 12 tubes of BGLB broth with the Durham gas collection tubes for the confirmed test; 2) Select only the positive lactose broth culture and transfer them to the BGLB using 2 times of inoculation loop; 3) Incubate all tubes for 24-48 hours at 370C; 4) After 48 hours, read the number of positive tubes for analyzing the MPN of total coliform bacteria as can be seen in table 12.2 Fecal Coliform Bacteria 1) Prepare the 12 tubes of EC-medium broth with the Durham gas collection tubes for the confirmed test; 2) Select only the positive lactose broth culture and transfer them to the EC-medium broth using 2 times of inoculation loop; 3) Incubate all tubes for 24-48 hours at 45.50C. 4) After 48 hours, read the number of positive tubes for analyzing the MPN of fecal coliform bacteria as can be seen in table 12.2 Lab III completed test 1) Transfer the positive of EC-medium using the inoculation loop in the EMB agar, then streak the loop on the EMB agar for E.coli isolation; 2) Incubate for 18-24 hours at 35.5 0C+5 0C; 3) Examine all of the plates that the E.coli colonies are blue-black with a metallic green sheen on the plates; 4) Identify the confirmative result of E.coli using the negative gram staining technique. 5) Record your result of sample that illustrate the E.coli contamination. Page | 125

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 10ml Swab or food sample LB 2X 1ml 0.1ml Presumptive Test LB 1X LB 1X Incubate 37 0C, 24-48 hr Gas fermentation in Durham Tube Confirmed Test EC-Medium BGLB Incubate 37 0C, 24-48 hr Incubate 45.5 0C, 24-48 hr Page | 126

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Confirmed Test EC-Medium Incubate 37 0C, 24-48 hr Incubate 45.5 0C, 24-48 hr Gas fermentation in Durham Tube Gas fermentation in Durham Tube Read the positive tube for finding the number Complete Test of total coliform bacteria using MPN table Streak the positive EC-Medium on EMB agar Incubate 35.5+5 0C, 18-24 hr Metallic Green Sheen Or E.coli colonies Page | 127

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Lab Result The Presumptive Test Sample Gas Reading MPN 95% Tube Probability Tube LB1X (1ml) Tube Range LB2X 123 LB1X (10ml) (0.1ml) 123 123 The Presumptive test (Positive and Negative tube) Name of sample……………………………………………………. Sample location…………………………………………………….. Page | 128

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory The Confirmed Test for total coliform bacteria Sample Gas Reading MPN 95% Tube Probability Tube BG1X-1ml Tube Range BG1X-10ml 123 BG1X-0.1ml 123 123 The Confirmed test for total coliform bacteria (Positive and Negative tube) Name of sample……………………………………………………. Sample location…………………………………………………….. Page | 129

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory The Confirmed Test for fecal coliform bacteria Sample Gas Reading MPN 95% Tube Probability Tube EC1X-1ml Tube Range EC1X-10ml 123 EC1X-0.1ml 123 123 Picture 12.3 The Confirmed test for Fecal coliform bacteria (Positive and Negative tube) Name of sample……………………………………………………. Sample location…………………………………………………….. Page | 130

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory The Completed test The complete test for E.coli Name of sample……………………………………………………. Sample location…………………………………………………….. The complete test for E.coli gram staining Name of sample……………………………………………………. Sample location…………………………………………………….. Page | 131

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Please identify the food sample and the location contaminated the E.coli found in the EMB agar Hand-swab Food container Food sample Utensil Sample Conclusion for the better of food sanitation and safety ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… ………………………………………………………………………………………… Page | 132

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 11.1 Chemical Contamination in food 11.1.1 Borax Borax, also known as sodium borate, sodium tetraborate, or disodium tetraborate, is an important boron compound, a mineral, and a salt of boric acid. Powdered borax is white, consisting of soft colorless crystals that dissolve easily in water. Its use as a cooking ingredient is to add a firm rubbery texture to the food, or as a preservative such as pork ball and ground pork. However, the importance symptom occurring from the increasingly intake of borax (less than 5 milligrams for children and less than 15 milligrams for adults) is referred to kidney insufficiency syndrome and the allergenic effect for gastrointestinal tract. Figure 11.1 Borax Source: G & A Warburtons Ltd, 2016. 11.1.2 Formalin Formalin is a clear aqueous solution of formaldehyde and methanol used especially as a preservative. Some of food including seafood, vegetable, and animal entrails may be contaminated with the formalin. Several syndrome is occurred if you intake the high level of formalin, for example, cancer, abdominal pain, unconsciousness symptom, and death. Figure 11.2 Formalin Source: Indiantmart, 2019. Page | 133

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 11.1.3 Sodium Hydrosulfite Sodium Hydrosulfite or Sodium Dithionite (NaHSO3) is a white crystalline powder with a weak sulfurous odor. It is the sodium salt of dithionous acid. Although it is stable under most conditions, it will decompose in hot water and in acid solutions. It is a chemical substance to bleach the whitening color on food such as bean sprouts, santol, and pickled bamboo shoots. The symptom occurring from the food contaminated with sodium hydrogensulfite is abdominal pain, hypotension, and death. Figure 11.3 Sodium Hydrosulfite Source: Indian chemical company, n.d. 11.1.4 Salicylic Acid Salicylic acid is a type of phenolic acid, and a beta hydroxyl acid (BHA). It is probably best known for its use as a key ingredient in topical anti-acne products. Moreover, the growth rate of microorganism such as mold can be prohibited using the salicylic acid. There are; unfortunately, several symptom if you intake the high level of salicylic acid including the fever, deficiency of immunization, tinnitus symptom, and death. Figure 11.4 Salicylic Acid Source: Hale Cosmeceuticals Inc, 2009. Page | 134

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 11.1.5 Polar Compounds Polar compound is definded as the fatty acid, monoglyceride, and triglyceride associated with the frequency of reuse on the cooked oil. In fact, the more you reuse the cooked oil, the more of cooked oil is oxidized, hydrolyzed, and polymerized. Therefore, the hypertension and the Coronary heart disease may be occurred if you intake the polar compound more than 25% per day. Figure 11.5 Cocked Oil 11.2 Chemical contamination testing for Food Sanitation and Safety 11.2.1 Borax 1) Grind the sample to the small size (approximately the head side of match) Source: กรมวิทยาศาสตร์การแพทย์, 2551. Page | 135

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 2) Bring 1 spoon of the sample to the cup of test-kit Source: กรมวิทยาศาสตร์การแพทย์, 2551. 3) Drop the test-kit solution, then mix the sample and solution Source: กรมวิทยาศาสตร์การแพทย์, 2551. 4) Dip the curcuma paper in the sample, then dry this paper around 10 minutes Source: กรมวิทยาศาสตร์การแพทย์, 2551. 5) Read the color on the curcuma paper, the red color is provided on the paper if the borax is contaminated in the sample (more than 50 milligrams per kilograms of sample) Page | 136

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Source: กรมวิทยาศาสตร์การแพทย์, 2551. 11.2.2 Formalin 1) Pour a liquid sample in bottle in NO.1 test-kit solution around 1/3 of bottle, or; 2) Use the distilled water soaked for the solid sample around 1-2 minutes, rinse the water-soaked sample in NO.1 test-kit solution around 1/3 of bottle, 3) Pour the sample from the previous step to the NO.2 test-kit solution, then mix and stir the sample, 4) Pour the sample from the previous step to the NO.3 test-kit solution, then mix and stir the sample immediately, 5) Read the color from the NO.3 test-kit solution, the red or pink color is provided if the sample is contaminated with the formalin. 11.2.3 Sodium Hydrosulfite 1) Pour the 5 ml of liquid sample into the test-kits cup; or Source: กรมวิทยาศาสตร์การแพทย์, 2551. 2) Use 10 ml of the distilled water soaked for the solid sample around 1-2 minutes, rinse the water-soaked sample in the cup Page | 137

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Source: กรมวิทยาศาสตร์การแพทย์, 2551. 3) Drop the 3 droplet of solution into the cup and then mix the sample Source: กรมวิทยาศาสตร์การแพทย์, 2551. 4) Read the color on the sample, the dark or gray color is demonstrated if the sodium hydrosulfite is contaminated in the sample. Page | 138

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 11.2.4 Salicylic Acid 1) Mark the label of two cups of test-kit including the NO.1 (the sample test) and NO.2 (the standard color of salicylic acid), and pour the 5 ml of liquid sample into the test-kits cup Source: กรมวิทยาศาสตร์การแพทย์, 2551. 2) Drop the 1 ml of the salicylic acid solution I into the NO.2 of cup only Source: กรมวิทยาศาสตร์การแพทย์, 2551. 3) Drop 1 ml of salicylic acid solution II for each cup, do not mix the sample for this step Page | 139

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 5) Read the color on the surface where; If the dark or violet color is provided on the NO.1 cup as same as the NO.2, this sample is contaminated with salicylic acid, Source: กรมวิทยาศาสตร์การแพทย์, 2551. If the dark or violet color is not provided on the NO.1 cup as same as the NO.2, this sample is not contaminated with salicylic acid. Source: กรมวิทยาศาสตร์การแพทย์, 2551. Page | 140

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory 11.2.5 Polar Compounds Test in the cooked oil 1) Drop the 4 droplets of Polar Compounds Test solution in the test tube Source: กรมวิทยาศาสตร์การแพทย์, 2551. 2) Drop the 2 droplets of cocked oil in the test tube Source: กรมวิทยาศาสตร์การแพทย์, 2551. 3) Mix and stir the sample for 30 seconds 4) Read the color on the sample, the pink color is demonstrated if the cooked oil contain with the polar compound in the standard (not over 25% of polar compound in cooked oil) Page | 141

Vivat Keawdounglek (Ph.D.) Learning material of 1807307 (Food Sanitation and Safety) Part 10 to 11: Principle of Food Sanitation Laboratory Laboratory Result for Chemical Contamination on Food Type of Type of food Source of food The Result Reference Picture chemical contamination 1. Borax 2. Formalin 3. Sodium Hydrosulfite 4. Salicylic Acid 5. Polar Compounds 6. (Other) Page | 142