EFFECTS OF CONTINOUS ADMINISTRATION OF LOW-DOSE OF Escherichia coli LIPOPOLYSACCHARIDE IN CHICKS AND POULTS FED NON TOXIC DOSES OF AFLATOXIN B1 AND T-2 TOXIN A Thesis presented to the Faculty of the Graduate School University of Missouri - Columbia In Partial Fulfillment of the Requirements for the Degree Master of Science by ELISÂNGELA APARECIDA GUAIUME Dr. D. R. Ledoux, Thesis Supervisor JULY 2005 The undersigned, appointed by the Dean of the Graduate Faculty, have examined the thesis entitled EFFECTS OF CONTINOUS ADMINISTRATION OF LOW-DOSE OF Escherichia coli LIPOPOLYSACCHARIDE ADMINISTRATION IN CHICKS AND POULTS FED NON TOXIC DOSES OF AFLATOXIN B1 AND T-2 TOXIN Presented by ElisAngela Aparecida Guaiume a candidate for the degree of MASTER OF SCIENCE and hereby certify that in their opinion it is worthy of acceptance. David R. Ledoux k& -& Alex I. Bermudez V yglLAA4- Trygve Veurn ACKNOWLEDGEMENTS I would like to thank my advisor, Dr. David Ledoux, for giving me the huge opportunity to work with him. Dr. Ledoux is a great advisor, very knowledgeable, approachable, and a great person to work with, always very patient, understanding and friendly. I could not have made it this far if it was not for Dr. Ledoux’s help since the first ten minutes I arrived in the States without knowing the language or anything else. Since that day, Dr. Ledoux has been a person I know I can count on. He encouraged me and has been of great assistance throughout my educational career. Dr. Ledoux, thank you very much for all your help! I appreciate everything you have done to help me to get this far. I also would also like to thank Dr. Alex Bermudez for his expertise in avian pathology and for his time answering many questions I had at the beginning of my program. Thanks for your patience. I also would like to thank Dr. Trygve Veum for being part of my committee. Thanks to Jonathan Broomhead for everything he taught me while working with him. No matter what we were doing or where we were, Jon always had something new to show me (including how to drive). It is going to be quite different not to have him around as he was always helpful even when I called him on Sundays. Thank you, Jon. Thanks to Carrie Walk for her enormous help not only with my experiments, but also in a not very easy moment of my life. Thank you for being so understanding, I will always appreciate it. I also would like to thank my dear friends Paula Butkeraitis and Leonardo Linares for their friendship and patience. This journey would have been so difficult if I did not have friends around. To Leonardo, I would like to thank him so much for his help and emotional support when I first came to the USA (and for teaching me how to drive). For Paula, I would like to say that she has been very important in my life and very helpful in all aspects. Her presence in my life is fundamental. Also, thanks to my dear friends Fabiana Farias and Lilian Pulz for having to put up with my complaints. Thanks to the friends that came and then left: Itamar and Bianca Martins. Thanks to the friends that ii were very important when I first came here: James Vuchetich, Eduardo, Sueli, Dalva, Christiane, Jim, Julie, Paulo, Emanuelle, Xavier, Vanessa, and Brainho. Thanks to my dear friend José Evandro de Moraes (Égon) who stayed in Brazil but, when possible, would call to encourage me when things did not seem to be working very well. Thanks to my young brother, Márcio José Guaiume, for his never-ending support regarding coming to the States, spending hours in line with me for consecutive days when I was applying for my student Visa and for his happiness when I got it. For me, his love was above everything else I was getting at that moment. I also would like to send special loving thanks to my parents, José Guaiume and Joanita Ferreira Guaiume for their enormous support every single day of my life and for being so proud of me for getting my Master’s Degree here in the States and for speaking my poor English (but they think my English is the best). They are always so proud of me. They started it all. The choice of leaving them was the hardest one ever, but they manage to stay always close even though we are some 4,000 miles apart. I love you!!! I also would like to thank my older brother, Marcos Paulo Guaiume, for his love and support and for my little nephews, Gustavo (I simply love spending time with this lovely child) and Guilherme (who, when I am with him, never knows that I am the voice he hears coming from the phone when I am apart). Besides my parents, leaving Gustavo was very difficult as he was a baby and I was afraid he was going to forget me. Lastly, I would like to express my appreciation to everybody that helped me starting, during, and terminating my dozens of experiments and to those who helped me with the never-ending injections: Jonathan Broomhead, Carrie Walk, Paula Butkeraitis, Fabiana Farias, Yvette Broomhead, Lilian Pulz, Itamar, Crystal Ledoux, Nicole Rasch, and Shannon. Also, thanks to my Lebanese friend Wassim Nasr for being so present, loving, understanding, and patient. To Jim Porter and Julie Sampson for their help with laboratory techniques I was not familiar with. I wish to thank Doris Lyons and Jesse Lyons for always being ready to help out with anything and Mary Smith for helping me tie up the loose ends. THANK YOU ALL!! iii TABLE OF CONTENTS ACKNOWLEDGEMENTS....................................................................................ii LIST OF TABLES ................................................................................................. v LIST OF FIGURES..............................................................................................vii ABSTRACT........................................................................................................viii Chapter I. INTRODUCTION.................................................................................................1 II. LITERATURE REVIEW.....................................................................................2 AFLATOXIN B1..........................................................................................2 T-2 TOXIN...................................................................................................15 LIPOPOLYSACCHARIDE ..........................................................................36 III. EFFECTS OF LIPOPOLYSACCHARIDE ON PERFORMANCE ON MORTALITY RATE, AND ORGAN WEIGHTS OF CHICKS AND POULTS FED AFLATOXIN B1 ..................................................................56 Abstract ........................................................................................................56 Introduction ..................................................................................................58 Material and Methods ...................................................................................59 Results ..........................................................................................................61 Discussion.....................................................................................................62 Conclusion....................................................................................................67 IV. EFFECTS OF CONTINUOUS ADMINISTRATION OF LOW-DOSE OF ENDOTOXIC LIPOPOLYSACCHARIDE IN CHICKS AND POULTS FED NON TOXIC DOSES OF T-2 TOXIN..................................................85 Abstract ........................................................................................................85 Introduction ..................................................................................................86 Material and Methods ...................................................................................90 Results ..........................................................................................................92 Discussion.....................................................................................................93 Conclusion....................................................................................................97 iv V. EFFECTS OF CONTINUOUS ADMINISTRATION OF LOW-DOSE ENDOTOXIC LIPOPOLYSACCHARIDE IN CHICKS AND POULTS FED NON TOXIC DOSES OF AFLATOXIN B1 AND T-2 TOXIN............101 Abstract ........................................................................................................101 Introduction ..................................................................................................102 Material and Methods ...................................................................................106 Results ..........................................................................................................108 Discussion.....................................................................................................111 Conclusion....................................................................................................120 VI. CONCLUSION ..................................................................................................150 REFERENCES.........................................................................................................152 v LIST OF TABLES Table Page 3.1. Effects of endotoxic lipopolysaccharide on 3 week performance of broiler chicks fed Aflatoxin B1 ............................................................................................71 3.2. Effects of endotoxic lipopolysaccharide on 3 week performance of turkey poults fed Aflatoxin B1.............................................................................................73