1st progress report Pauisti These PhD.pptx

Project these for PhD

1. 1 Detection, Identification and Quantification of aflatoxin producing fungi using loop-mediated isothermal amplification (LAMP) assays, and evaluation of biocontrol agents on growth of Aspergillus strains from rice grains in Kenya DOUKSOUNA Youmma MB: 400-0001/17 PhD candidate in Molecular Biology and Biotechnology Supervisors Dr. STEVEN Runo / Kenyatta University Prof. AMBANG Zachée  / University of Yaoundé 1 Dr. ANDREW K. Nyerere / JKUAT

2. 2 I. BACKGROUND OF THE STUDY II. PROBLEM STATEMENT III. OBJECTIVES IV. Collection and processing of samples. V. Moisture content and selection of suspected grains outline

3. I. BACKGROUND OF THE STUDY 3 Nearly every food or feed commodity can be contaminated by fungal pathogens Mycotoxins can develop inside some food when environmental conditions are favorable and lead to biosynthesis of Aflatoxins Rice ( Oryza sativa L .) is one of the most important food consumed in sub Saharan Africa It is considered an appropriate substrate for fungal growth (Lai et al., 2015) Rice is often contaminated with mycotoxins such as aflatoxins; Aflatoxins B 1 , B2, G1, and G2 are most significant contaminants to rice ( Su´arez -Bonnet E., et al., 2013)

4. II. PROBLEM STATEMENT 4 Murphy et al., 2006 Williams, J.H.; et al., 2004 Figure 2. Distribution of Hepatocellular Carcinoma Attributable to Aflatoxin (Liu and Wu, 2010) Examples Cereals ( maize, rice, wheat ); Oil seeds (groundnut, soybean, sunflower, cotton); species (black pepper, coriander, turmeric, zinger); tree nuts (almonds, pistachio, walnuts, coconut); cows / others (Milk, meat; eggs)

5. II. PROBLEM STATEMENT con’t The following effects may occur singly or in combination 5 World health authorities warn that low doses with long-term dietary exposure to aflatoxins is a major risk to lead hepatocellular carcinoma ( Marta T., et al 2016 )

6. 6 General objective To adapt loop-mediated isothermal amplification (LAMP) assays for detection and identification of aflatoxin producing fungi, and evaluate biocontrol agents (BCAs) on the growth of Aspergillus strains on rice grain in vitro . Specific Objectives 1. To isolate, identify and characterize Aspergillus strains using microbial culture techniques 2. To adapt LAMP for detection of Aflatoxin producing by Aspergillus species isolated from rice grains and quantify aflatoxins levels by using validated methods, 3. To characterize expression of genes responsible for producing aflatoxins by Aspergillus strains 4. To evaluate efficacy of biocontrol agents by competitive exclusion on growth of Aspergillus strains in vitro III. OBJECTIVES

7. 7 IV. Collection and processing of samples. Sample size was estimated to Samples. Samples of rice grains ( white,and brown) was collected randomly from different sales points Mwea , Thika , Ahero , Kisumu and Nairobi . Sampling method was done according to the sampling protocol for Official Control of Mycotoxins in Food (EU, 2003) to give a representative sample which was then placed in sealed bags.  

8. 8 I conducted a field work collection of 98 samples of different varieties of rice grains namely Pishori , Sindano , Brown, Parboiled, Pakistan, Thailand, Biryani and India at Mwea and Thika. A. Rice stores and Rice Millers in Mwea A B

9. 9 C C and D imported rice (Pakistan, Thailand, India, Biryani, Parboiled) in Thika   D

10. 10 F. Sample suspected contaminated with discoloration

11. 11 V. Moisture content and selection of suspected grains

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16. THANK YOU FOR YOUR KIND attention