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  3. Investigating the possibility of binding aflatoxin M1 to Lactobacillus rhamnosus GG and Bifidobacterium lactis BB12 in ice cream

Investigating the possibility of binding aflatoxin M1 to Lactobacillus rhamnosus GG and Bifidobacterium lactis BB12 in ice cream

Number of pages: 104 File Format: word File Code: 32411
Year: 2014 University Degree: Master's degree Category: Food and Packaging Industries
Tags/Keywords: Aflatoxin - Aflatoxin binding - Aflatoxin M1 - BB12 - Bifidobacterium lactis - Dairy products - ice cream - lion poison - Mycotoxin - Probiotic - Rhamnos GG
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  • Summary of Investigating the possibility of binding aflatoxin M1 to Lactobacillus rhamnosus GG and Bifidobacterium lactis BB12 in ice cream

    Dissertation for Master Degree in Agricultural Engineering,

    Food Science and Industry

    Tension: Food Microbiology

    Abstract

    Aflatoxin M1 (AFM1) is one of the important mycotoxins found in milk and dairy products and is a metabolic product of aflatoxin B1. Today, it is possible to remove mycotoxins in food or animal feed by using physical, chemical or microbial methods. The aim of this research is to find a safe and harmless way to remove aflatoxin in the milk probiotic product, which does not pose any nutritional risk to humans. In this study, ice cream samples were prepared from reconstituted milk without primary aflatoxin. In all samples (control and probiotic), the changes of aflatoxin during the process were checked at two ppb levels (0.05, 0.1). Growth changes of Lactobacillus rhamnosus GG and Bifidobacterium lactis 12Bb were studied in aerobic and anaerobic MRS medium at 37°C. HPLC method was used to measure aflatoxin in ice cream samples. For this purpose, 20 samples with concentrations of aflatoxin M1 (0.05, 0.1 ppb) and 30% acetonitrile were used as solvents. The results of the statistical analysis showed that the amount of aflatoxin reduction at the end of the heating stage was 22.4% and 29.5% for the concentrations of 0.05 and 0.05 ppb, respectively, and there was a significant decrease (p<0.01). At the end of the fourth week of ice cream storage, the amount of aflatoxin reduction in both ppb levels (0.1, 0.05) which were 30.6% and 32.1%, respectively, had no significant difference (p<0.01). The microbial population growth of Lactobacillus rhamnus GG and Bifidobacterium lactis 12BB increased significantly (p<0.05) in all three samples during the warm house stage. The logarithm of the microbial population of Lactobacillus rhamnus GG at the end of the fermentation stage for all three concentrations of aflatoxin M1 ppb (0.05, 0.1) is 10.132, 10.02, 10.01 CFU, and for Bifidobacterium lactis 12 BB, it is 9.30, 9.30, 9.00 CFU, respectively. Also, the concentration of aflatoxin had a significant effect (p<0.05) on the growth of the microbial population of both species. During storage, the reduction of the logarithm of the microbial population for both types of probiotics in the higher concentration of aflatoxin was not significantly different from the samples containing the lower concentration of aflatoxin (p<0.05). M1 (AFM1) is one of the important mycotoxins found in milk and dairy products. Today, it is possible to remove mycotoxins in food or animal feed by using physical, chemical or microbial methods. The aim of this research is to find a safe and harmless way to remove aflatoxin in the milk probiotic product, which does not pose any nutritional risk to humans. In 2005, milk production in the world was estimated at 644 million tons, of which 541 million tons were cow's milk. By looking at the above statistics, we can understand the health issue of the milk consumed and consequently the dairy products produced from this amount of milk. The increasing consumption of dairy products by different people around the world has become a challenging topic for microbiologists, engineers, and technologists; Because they should find the best way to prevent the entry of microorganisms and destroy some of them and prevent their growth and activities. However, the issue that should be considered is that many of these microorganisms such as lactobacillus and bifidobacteria are beneficial for human health, and others, such as some fungi such as Aspergillus, which produce aflatoxin, can be harmful to human health. Milk contains a large amount of water and nutrients and provides an almost neutral environment, so milk can be considered as a source of toxic compounds such as mycotoxins (Maqsoudi 2019).

    Also, the nutritional nature of dairy products provides a suitable environment for the growth of microorganisms. Aflatoxin M1, known as milk toxin, is the main metabolite formed from AFB1. AFM1 can be found in the human body 12 hours after consuming AFB1. Due to the widespread consumption of milk and dairy products, the presence of aflatoxin M1 in these products has become a big concern. Therefore, finding out how milk is contaminated with contaminating microorganisms is one of the most important measures in dealing with the health of milk and then dairy products. Currently, milk consumption in Iran is 70 to 80 kg, which is 350 to 500 kg in developed countries. Therefore, the need to produce a quality product as a supplier of part of the calcium and protein consumption can answer some of the nutritional problems of the society. Milk, which is considered as a complete food, has many products that are consumed by the general public, and ice cream is particularly important as one of the important products of Shiraz. In the past years, the consumption of ice cream was mostly reserved for children and teenagers, but now its consumption has spread to all members of the society, and of course, the production and consequently the consumption of all kinds of ice cream has grown considerably. In milk, there are agents that produce aflatoxins; Therefore, it is very important to provide conditions that can reduce the amount of this poison and provide a healthier product. Considering the importance of aflatoxin in carcinogenesis and reducing human immunity and health, the purpose of this study is to investigate the amount of aflatoxin in ice cream obtained from milk, and since the consumption of ice cream, which is considered a probiotic product of milk, is of particular importance in terms of improving human health, it seems necessary to carry out this research study.

    1-4- Specific research objectives

    and without harm to remove the aflatoxin poison in the milk lactic product, which does not pose any nutritional risk to humans.

    Investigation of the effect of lactic bacteria on different amounts of aflatoxin in probiotic ice creams using HPLC method.

    Determination of the effect of milk acidity on different amounts of aflatoxin.

    Evaluation of the effect of different periods of storage on different amounts of aflatoxin.

     

     

    1-5- Hypotheses

    Using Lactobacillus rhamnos GG and Bifidobacterium lactis in the production of probiotic ice cream reduces the amount becomes aflatoxin.

    Lactic bacteria can affect different amounts of aflatoxin.

    The amount of different amounts of aflatoxin can affect lactic bacteria.

    The amount of milk acidity affects the activity of lactic bacteria in different amounts of aflatoxin. It is significant.

    Different storage times have a significant effect on different amounts of aflatoxin.

  • Contents & References of Investigating the possibility of binding aflatoxin M1 to Lactobacillus rhamnosus GG and Bifidobacterium lactis BB12 in ice cream

    List:

    Chapter One: Research overview. 2

    1-1- Introduction. 3

    1-2- statement of the problem. 3

    1-3- Necessity of doing research. 4

    1-4- specific objectives of the research. 4

    1-5- Hypotheses 5

    1-6- The main research question. 5

    1-7- The innovation aspect of research. 5

    Chapter two: Review of research literature. 6

    2-1- Definitions. 7

    2-1-1- History of ice cream. 7

    2-1-2- Definition of ice cream. 7

    2-1-3- per capita consumption of ice cream and milk. 8

    2-1-4- The role of each ingredient used in ice cream. 9

    2-1-4-1- fat. 9

    2-1-4-2- Solids without fat (MSNF) 9

    2-1-4-3- Sugar. 9

    2-1-4-4- Emulsifiers 9

    2-1-4-5- Stabilizers 9

    2-1-4-6- Flavorings 10

    2-1-4-7- Inulin. 10

    2-1-5- Probiotics 10

    2-1-5-1- History of the use of probiotics 10

    2-1-5-2- Probiotic definition. 11

    2-1-5-3-properties of probiotics. 12

    2-1-5-4- The most important productions of probiotics 12

    2-2- Review of research background. 13

    2-2-1- Fermented milk. 13

    2-2-2- Probiotics 13

    2-2-2-1- Therapeutic and preventive effects of probiotics 13

    2-2-2-2- the most important probiotics used. 15

    2-2-2-3- Side effects of taking probiotic lactobacilli. 15

    2-2-2-4- beneficial effects of probiotic bacteria. 16

    2-2-2-4- Probiotic products. 17

    2-2-2-5- dominant bacteria in probiotic ice cream. 17

    2-2-2-6- Lactobacillus ecology 18

    2-2-3- Probiotic ice cream. 19

    2-2-3-1- types of probiotic ice cream. 19

    2-2-3-2- obstacles to the production of probiotic ice cream. 20

    2-2-3-3-Methods to remove obstacles to the production of probiotic ice cream. 20

    2-2-4-mycotoxins 20

    2-2-4-1-effective factors in the growth of fungi and mycotoxin production in food. 21

    2-2-5- Aflatoxins 24

    2-2-5-1- A brief review of the history of aflatoxin. 26

    2-2-5-2- Pathology of aflatoxins 27

    2-2-5-3- Microbiology of aflatoxins 28

    2-2-5-4- Fungi producing aflatoxins 29

    2-2-5-5- International standard rules. 30

    2-2-5-6- Global limits of aflatoxins in nutrition. 31

    2-2-5-7- methods to reduce aflatoxin: 32

    2-2-5-8-aflatoxin in milk. 33

    2-2-5-9-Evaluation and control of pollution. 35

    2-2-5-10- Testing methods. 35

    2-2-6-high performance liquid chromatography. 36

    2-2-6-1- HPLC device. 37

    2-2-6-2- detector 38

    2-2-6-3- measurement methods. 39

    2-2-6-4- HPLC solvents. 39

    2-2-6-5- Preparing and working with the device. 40

    2-3- An overview of past research. 41

    2-4- Biodegradation of mycotoxins, current situations and future approaches. 52

    2-4-1- Removing pollution using fermentation. 52

    Chapter three: materials and methods 58

    3-1- Basic work 59

    3-2- Materials used in making ice cream. 61

    3-3- Equipment and devices used in ice cream preparation and tests. 62

    3-4- Materials and equipment needed to measure aflatoxin M1 63

    3-5- Methods 65

    3-5-1- Research variables. 65

    3-5-2- Tests related to powdered milk. 65

    3-5-3- Tests performed on ice cream syrup during fermentation. 66

    3-5-3-1- pH determination test. 66

    3-5-3-2- Microbial population growth test 66

    4-2- Evaluation of pH changes in the greenhouse stage. 74

    4-3- Evaluation of the effect of aflatoxin M1 concentration on changes in the growth of Lactobacillus rhamnus GG microbial population in the greenhouse stage. 77

    4-4- Evaluation of the effect of aflatoxin M1 concentration on changes in the growth of the microbial population of Bifidobacterium lactis 12BB in the greenhouse stage. 79

    4-5- Evaluation of changes in aflatoxin M1 during heating and greenhouse. 82

    6-4 Evaluation of the effect of aflatoxin M1 concentration on changes in the growth of the microbial population of Lactobacillus rhamnus GG during storage. 87

    4-7- Evaluation of the effect of aflatoxin M1 concentration on changes in the growth of the microbial population of Bifidobacterium lactis 12BB during the storage period. 88

    4-8- Evaluation of changes in aflatoxin M1 during storage. 91

    The fifth chapter:91

    Chapter five: general conclusions and suggestions. 92

    5-1- General conclusion. 93

    2-5- Suggestions. 94

    List of sources. 95

    Appendices 106

    Source:

    Ebrahimi Tajabadi M. 1388. Identification of probiotics in Iran's dairy products. PhD thesis, Department of Food Industry, Faculty of Agricultural Engineering, Islamic Azad University, Science and Research Unit.

    Benonym, National Standard No. 11325, (1390), Publications of Iran Institute of Standards and Industrial Research, Probiotic Yogurt - Characteristics and Test Methods, First Edition.

    Benonym, National Standard No. 14683, (1389), Publications Iran Standard and Industrial Research Institute, Probiotic ice cream, characteristics and test methods, first edition.

    Banonym, National Standard No. 2450, (2004), Publications of Iran Standard and Industrial Research Institute, Ice cream - characteristics and test methods, fifth revision.

    Tabari M. and Amin Afshar M. 2018. Investigating the level of aflatoxin-M1 contamination of milk and yogurt and the effect of the production process on the level of aflatoxin in Gilan province. The final report of the plan approved by the Islamic Azad University of Lahijan branch number 116. Tawakli H. 1389. Investigating contamination with aflatoxin M1 in pasteurized cheese supplied by two factories producing dairy products in Tehran using the Eliza method.

    Chegani B, Mashkoh A, ice cream knowledge and technology, Ayiz publisher, Tehran, first edition 1385

    Khaksar R, Karim G, Ferdowsi R. 1385. Validation of the test method for measuring aflatoxin M1 in Iranian white cheese by HPLC. Veterinary Journal 4:21-34.

    Khosravi Darani K, Kushki M. 2017. Probiotics in milk and its products. Tehran: Merz Danesh Publications, 268 pages.

    Khoshnavis et al. 2012. Determination of aflatoxin M1 in ice cream in Babol, North of Iran.

    Razzaghi Abianeh M, Ahmadi Y, Ghafarakhi M, Alinejad S. 1390. Aflatoxins and their importance in public health and agriculture. Tehran. First. Publications: Jihad Agricultural Applied Scientific Higher Education Institute, 209 pages.

    Rohi P, Nikopour H, Farahani A, Khosravi Darani K. 1385. The effect of process variables on the production of conjugated linoleic acid in probiotic yogurt by Takoji method, 16th National Congress of Food Industry, Safety, Waste Reduction, Innovation, Gorgan University of Agriculture and Natural Resources, Gorgan, Iran.

    Salari R, Najafi MB. 2018. Comparison of ELISA and HPLC methods in determination of aflatoxin B1 and acratoxin A in Iranian red pepper. Journal of Food Industry Research, Volume 21, Number 4: 5-16.

    Tahari P, Ehsani MR, Khosravi Darani K, Razavi S. 1385 A. Evaluation of parameters affecting acidification and pH reduction of probiotic yogurt containing Lactobacillus La-5 during the greenhouse period, Journal of Food and Nutrition Industries, (1) 4, 37-46.

    Tahari P, Ehsani M, Khosravi Darani K, Razavi S. 2015. Evaluation of different culture media for enumeration of Bifidobacterium genus as probiotic bacteria, 16th National Congress of Food Industry, Security, Waste Reduction, Innovation, Gorgan University of Agriculture and Natural Resources, Gorgan, Iran.

    Tahari P, Ehsani M, Khosravi Darani K, Razavi S. 1385. The effect of primary milk composition, percentage and type of inoculation on the microbial growth of probiotic yogurt containing Lactobacillus La-5 during the greenhouse period, Isfahan University of Technology, Isfahan, Iran. Evaluation of process variables on the production of probiotic drink yogurt, master's thesis, Department of Food Industry, Faculty of Agricultural Engineering, Islamic Azad University, Science and Research Unit. 2018. Production of fermented drink based on whey using different types of kefir microflora and investigating its chemical and organoleptic characteristics. Iranian Journal of Nutritional Sciences and Food Industries 4: 21-32.

    Karim, G., Milk and its products, Sepehr Publishing House, Tehran, second edition, 1380

    Lame, H., Bardgan, Smith, Jim, Guide to the use of food additives, Islamic Azad University Scientific Publishing Center, Tehran, first edition, 1374

    Morhamatizadeh MH, Rezazadeh S, Jafari A. 2018. Production of probiotic ice cream using cultures of Lactobacillus acidophilus and Bifidobacterium bifidum. New Veterinary Research Journal 5: 33-39.

    Moeinian K, Yaghmaian K. 2012. Aflatoxin M1 concentration in raw milk produced in the cities of Semnan province. Kumesh Magazine 15:21-27.

    Maqsodish, Ice Cream Production Technology, Iranian Agricultural Science Publication, Tehran, 1390

    Mektabis S, Haji Kalaei MR, Najafabadi M. 1390.

Investigating the possibility of binding aflatoxin M1 to Lactobacillus rhamnosus GG and Bifidobacterium lactis BB12 in ice cream
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