VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE NGUYEN THI THANH XUAN ASSESSMENT OF AFLATOXIN B1 CONTAMINATION IN MAIZE AND AWARENESS OF AFLATOXINS IN SON LA PROVINCE Major : Food Science and Technology Code : 60540103 Supervisor : 1. Hu Suk Lee 2. Nguyen Thi Thanh Thuy AGRICULTURAL UNIVERSITY PRESS - 2017 LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com DECLARATION This thesis contains no materials that have been accepted for the award of any other degrees or diploma in any educational institutions. To the best of my knowledge, my paper includes no previously published work written by any person, except where indicated in references.
Hanoi, May 10th, 2017 Master candidate Nguyen Thi Thanh Xuan i LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com ACKNOWLEDGEMENTS First of all, I am really grateful my supervisors Dr. Hu Suk Lee from International Livestock Research Institute (ILRI) for his patience, enthusiasm, motivation and immense knowledge, and to Dr. Nguyen Thi Thanh Thuy from Vietnam National University of Agriculture, who gave me the opportunity to carry out this thesis and always had a piece of advice ready regarding the practical aspect of my work. Their guidance supported me during this research.
I wish to express my gratitude to Prof. Nguyen Viet Hung, a Southeast Asia regional representative at ILRI in Vietnam, for giving me a profound insight into the research and sharing his valuable experience with me. I always bear in mind the enthusiasm Prof. Delia Grace and Dr.
Johanna Lindahl at ILRI in Nairobi. I highly appreciate Ms. Nguyen Le Thanh and Ms. Le My Hanh for their help in the administrative aspect.
Eventually, I would like to thank to all the members at ILRI in Vietnam for giving me the opportunity to become temporarily a part of their team. I owe my thanks to all staffs in Plant Protection Research Institute. They helped me so much for conducting sampling, interview, data collection and laboratory analysis. Many thanks particularly to the whole team, who went to Son La for their logistical support and suggestion.
I also express my admiration to all members in Son La who introduced me to the locals and supervised our team during sampling and interview process. I am also very thankful to Son La residents who warmly welcomed our team in their homes and participated in the survey. Last but not least, I want to convey my gratefulness to my family and friends for their unremitting support, patience and understanding. Hanoi, May 10th, 2017 Master candidate Nguyen Thi Thanh Xuan ii LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com LIST OF CONTENTS Declaration.
ii List of contents. iii List of abbreviations. v List of tables. vii List of figures.
viii Thesis abstract. Situation in the maize production in vietnam – case of Son La. Situation in the maize production in Vietnam. Study location - Son La province.
Aflatoxins and their impacts on human and animal health. The impact of aflatoxins on human and animal health. Factors influencing aspergillus spp. infection and aflatoxin development in maize.
Situation in the aflatoxin contamination in maize. Situation in the aflatoxin contamination in the world. Situation in the aflatoxin contamination in Vietnam. Methods to detect aflatoxins.
Regulations for aflatoxins in the world and Vietnam. Regulations for aflatoxins in the world. Regulations for aflatoxins in Vietnam. Materials and methods.
The Sigma-Aldrich® Aflatoxin B1 Low Matrix ELISA kit. 24 iii LUAN VAN CHAT LUONG download : add luanvanchat@agmail. Sample size calculation and selection. Aflatoxin B1 analysis in Lab.
Survey data analysis. Results and discussions. Aflatoxin B1 contamination in maize in Son La. Awareness of aflatoxins in Son La.
Conclusions and recommendations. 58 iv LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com LIST OF ABBREVIATIONS Acronym Abbreviations A. flavus Aspergillus flavus A. nomius Aspergillus nomius A.
parasiticus Aspergillus paraciticus AFB1 Aflatoxin B1 AFM1 Aflatoxin M1 AFs Aflatoxins CIT Citrinin DON Deoxynivalenol EC European Commission EFTA European Free Trade Association ELISA Enzyme Linked Immunosorbent Assay EU European Union F. moniliforme Fusarium moniliforme FAO Food and Agriculture Organization of the United Nations FB1 Fumonisin B1 FDA Food and Drug Administration FUM Fumonisins GAPs Good agricultural practices GPS Global positioning system HBV Hepatitis B virus HCC Hepatocellular Carcinoma HPLC High-performance liquid chromatography IARC International Agency for Research on Cancer LOD Limit of detection v LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com OD Optical density OTA Ochratoxin A PBS Phosphate-buffered saline PE Polyethylene ppb Part per billion TLC Thin-layer chromatography USA United States of America ZEA Zearalenone µg/kg Microgram per kilogram vi LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com LIST OF TABLES Table 2. Area under maize, maize production and yield of districts in Son La in 2013. The property of aflatoxins.
Limits of aflatoxin growth and production by A. The FDA action levels for aflatoxins in human food and livestock feed. Vietnamese limits for aflatoxins in human food. Awareness of moldy contamination in maize in Son La.
Prevalence of AFB1 contamination in maize in Son La. Awareness of molds among 107 respondents in each category corresponding questions with answers. Practices associated with consumption of maize, meat and milk. Practices related to the medical examination.
45 vii LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com LIST OF FIGURES Figure 2. Agro-ecological regions in Vietnam. The administrative map in Son La province. Area under maize, maize production and yield in Son La province.
Chemical structure of aflatoxins. The map shows sampling sites in Son La province. Difference in the prevalence of AFB1 contamination in maize. Socio- demographic characteristics of respondents.
37 viii LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com THESIS ABSTRACT Master candidate: Nguyen Thi Thanh Xuan Thesis title: Assessment of aflatoxin B1 contamination in maize and awareness of aflatoxins in Son La province Major:Food Technology Code: 60540103 Educational organization: Vietnam National University of Agriculture (VNUA) Research Objectives This research was conducted to evaluate the prevalence of aflatoxin B1 contamination in maize and residents’ awareness of aflatoxins in Son La province. Materials and Methods Materials included maize samples and the Sigma-Aldrich® Aflatoxin B1 Low Matrix ELISA kit. Maize samples were randomly collected at the end of the rainy season from 25 communes in 5 districts in Son La with recording GPS location. All maize samples were analysed for AFB1 contamination in Lab, using the Sigma-Aldrich® Aflatoxin B1 Low Matrix ELISA kit.
Besides, questionnaire survey was simultaneously carried out with maize collection using face-to-face interviews. Eventually, all data were imported into Microsoft Excel 2010 and analyzed using STATA (version 14.0, StataCorp, College Station, TX, USA).1 ArcMap (ESRI, Redlands, CA, USA) was used to create the map. Main findings and conclusions The prevalence of AFB1 contamination in maize in Son La was lower when harvested in the field; however, this prevalence was higher in the rainy season. Noticeably, most people had adequate awareness of molds and their impacts on human and animal health, but they lack insight of aflatoxins and the impacts of aflatoxins, particularly ethnic minority groups.
In conclusion, the findings can conclude that maize in Son La is exposed to AFB1 to varying degree, and there may be a risk that maize production could contain AFB1. Therefore, we believe that this study produces evidence on potential risk to humans and animals in Son La and will inform science-based messages for reducing risk. Addtionally, the findings underline the need to enhance public's awareness about aflatoxins and health risks associated with aflatoxins on human beings and animals. ix LUAN VAN CHAT LUONG download : add luanvanchat@agmail.
INTRODUCTION Vietnam has an extremely advantageous geographical location in Southeast Asia and Asia. It is located in both tropical and sub-tropical areas. Thus, its climate features a tropical monsoon climate with high temperature and humidity which is a favorable condition for fungal growth, especially mycotoxins produced from Aspergillus spp. Among mycotoxins, aflatoxins, particularly aflatoxin B1 (AFB1) produced primarily by two species of ubiquitous fungus Aspergillus (A.
parasiticus have attracted general public attention because it is the most toxic and potent carcinogen even in small quantities (FAO, 1979 and De Campos, 1987). According to The International Agency for Research on Cancer (IARC), AFB1 is classified as group I carcinogens (Humans et al. Epidemiologically, AFB1 can lead to be genotoxicity and carcinogenic toxicity for both human and animal, especially the development of liver cancer (hepatocellular carcinoma) and higher risk to hepatic B virus infected individuals (WHO, 1998; Hansen et al. In fact, AFB1 is very susceptible in nuts and cereal, especially maize and groundnut during the harvest, or gradually susceptible while storing and processing (Reddy et al.
Therefore, it is extremely important to detect aflatoxins in food and animal feed for assurance of human and animal health as well as food hygiene and safety. In Vietnam, maize is the second only to rice as staple food for the consumption of humans and animals, especially in the rural and mountainous areas. It is regarded as the major energy feed source for Vietnam's livestock industry (Ha et al. Mountainous North-west provinces, such as Son La, are still listed as belong to the poorest in Vietnam with limited development opportunities over the past years.
Therefore, maize is also considered as one of the most important crops. It grows in different ecological regions to alleviate poverty in the provinces with difficult economic conditions. Nevertheless, maize was reportedly extremely good substrate for fungal development producing mycotoxins, especially aflatoxins (Wang and Liu, 2006 and Zinedine et al. Meanwhile, Son La province is the largest maize producing area in Vietnam but few comprehensive AFB1 investigations had been carried out to 1 LUAN VAN CHAT LUONG download : add luanvanchat@agmail.com establish the prevalence of AFB1 contamination in maize in Son La.
Previously, some studies on the occurrence of AFB1 in maize in Vietnam have provided some insight; however, these were based on small sample sizes or limited-scale geographical areas or relied on the data from one point in the maize value chain. For instance, AFB1 was surveyed in 32 maize samples in a Wang et al.’s study (1995), Trung et al. (2008) only conducted in 25 maize samples whereas a bigger sample size (97 samples) were researched by Thieu et al. (2008) and Phuong et al.
(2015) (Wang et al., 1995; Trung et al., 2008; Thieu et al., 2008 and Phuong et al. This would be difficult to generalize the population. Moreover, few comprehensive AFB1 contamination research to establish the prevalence of AFB1 contamination in maize and unique study about perception and knowledge of aflatoxins had been conducted in Son La (Lee et al. Therefore, an investigation extended greater sampling was conducted to assess AFB1 contamination in maize and awareness of aflatoxins in Son La province.
RESEARCH OBJECTIVES General objectives This research conducted is to evaluate the prevalence of aflatoxin B1 contamination in maize and residents’ awareness of aflatoxins in Son La province. Specific objectives - To evaluate the prevalence of aflatoxin B1 in maize in Son La province in an attempt to give recommendations to mitigate moldy contamination in maize. - To evaluate the awareness of aflatoxins among people so as to improve their understanding of health risks for both human beings and animals relating to aflatoxins contamination by consuming maize in Son La. 2 LUAN VAN CHAT LUONG download : add luanvanchat@agmail.
SITUATION IN THE MAIZE PRODUCTION IN VIETNAM – CASE OF SON LA 2. Situation in the maize production in Vietnam As in most of the Asian countries like China, Thailand, Indonesia, Vietnam, maize production have been dramatically increasing during the last decade along with the rapidly increasing demand for animal feed. The Vietnamese maize production boosted from 1,177.2 thousand tons in 1995 to 2,005.9 thousand tons in 2000 and drastically increased 5,281.0 thousand tons in 2015 (GSO, 2017). This rapid increase leads to both expansion in area and yield.