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i EVALUATION OF POTENTIAL VECTOR CONTROL METHODS THAT TARGET OUTDOOR FEEDING ANOPHELES MOSQUITOES MARY MATHEW MATHANIA A THESIS SUBMITTED IN FULFILMENT OF THE REQUIREMENTS FOR AWARD OF THE DEGREE OF DOCTOR OF PHILOSOPHY FOR SOKOINE UNIVERSITY OF AGRICULTURE. MOROGORO, TANZANIA 2017 ii ABSTRACT Insecticide-Treated Nets (ITNs/Long Lasting Insecticide-Treated Nets (LLINs) and Indoor Residual Spraying (IRS) have contributed to halving global malaria incidence and mortality rates. Achieving zero malaria transmission using these methods which target indoor-feeding mosquitoes is hampered by residual malaria transmission attributable to outdoor-feeding mosquitoes. This study evaluated potential malaria vector-control methods that target outdoor-feeding Anopheles mosquitoes to contribute to efforts to eliminate residual malaria transmission. Specifically, it set out: (i) To assess community knowledge and awareness on malaria and its vectors, (ii) To determine spatial distribution of Anopheles adults and larvae, (iii) To determine effects of combined use of chemo-attractants and non excito-repellant insecticide (Chlorfenapyr) applied to contaminating devices in attracting and killing outdoor Anopheles mosquitoes and (iv) To evaluate method of attracting gravid Anopheles gambiae s.s to oviposit in artificially-created ovicidal breeding sites. To meet specific objective (i), a cross-sectional questionnaire study which explored knowledge and awareness on malaria was carried out in Dodoma and Morogoro on 400 respondents. The questionnaire study revealed that a vast majority (78.8%) were not aware that early outdoor biting of mosquito was a risk factor for malaria transmission. The main Anopheles breeding sites were found to be rice paddies (25.2%), ditches (23.3%) and septic tanks/pits (18.8%). Adult Anopheles mosquitoes were collected mainly from ceiling (42.3%) and stored/piled junks (16.3%) in the hot-wet season and from under beds (32.1%), undisturbed curtains (25.3%) and store rooms (23.7%) in cold-dry seasons. On testing a novel Umbrella-topped Mosquito Contaminating Device (UtMCD) developed in the present studies as a tool for attracting and killing outdoor Anopheles mosquitoes (specific objective iii) it was revealed that, adjusting for season and study location, the number of Anopheles mosquitoes caught were significantly associated with type of UtMCD set-up used. From among the UtMCD iii set-ups A-D and device E tested (A: Device alone, B: Device with attractants alone, C: Device with insecticide alone, D: Device with insecticide and attractants and E: Device (Okomu), UtMCD set-up D significantly caught more Anopheles mosquitoes than UtMCD set-up A (AMR=2.96,p<0.0001). Mortality analysis showed that UtMCD set-ups D, C and device E had higher percentage mortality: D (87.7%); C (89.4%) and E (84.9%) than UtMCD set-up A (19.2%) and UtMCD set-up B (17.4%). In the studies on artificially-created breeding sites it was found that the odds of not hatching for eggs deposited in hay infusion +vinegar sites was significantly greater than that of eggs deposited in water alone (AOR=80.6, p<0.001). In conclusion, the studies revealed there is a need for increased awareness on malaria transmission by outdoor early-feeding Anopheles and furthermore, the novel UtMCD (with chemo-attractants and insecticide) as well as artificial breeding sites with attractants and an ovicidal agent (vinegar) have potential for use in integrated vector and larval source management to eliminate residual malaria transmission. iv DECLARATION I, Mary Mathew Mathania, do hereby declare to the Senate of Sokoine University of Agriculture that this thesis is my own original work done within the period of registration and that it has neither been submitted nor being concurrently submitted to any other institution. ___________________________ __________________________ Mary Mathew Mathania Date (PhD Candidate) The above declaration confirmed _______________________________ ___________________________ Prof. Richard S. Silayo Date (Supervisor) v COPYRIGHT No part of this dissertation may be reproduced, stored in any retrieval system, or transmitted in any form or by any means without prior permission of the author or Sokoine University of Agriculture in that behalf. vi ACKNOWLEDGEMENT Firstly I thank God Almighty for His Love and continual blessing. He is a well-spring of guidance and strength to me My sincerest gratitude goes to my supervisor, Professor Richard S. Silayo for tireless efforts in guidance, contributions and encouragement towards completion of this thesis. He was always inspirational and a wonderful role model being not only an awesome supervisor but also like a father and a friend to me. I really enjoyed his supervision. My gratitude also goes to Sokoine University of Agriculture. I appreciate all I have learnt from Sokoine University of Agriculture and the support I got from all staff. I am also deeply grateful to Prof. Sharadhuli I. Kimera for his detailed constructive criticism and invaluable contribution to this work. Additionally, I am deeply indebted to Prof. Elikira Kimbita the former and Prof Gerald Misinzo the current Head of Department of Veterinary Microbiology and Parasitology (currently Department of Microbiology, Parasitology and Biotechnology of College of Veterinary Medicine and and BioMedical Sciences) as they sacrificed much to see this work become successful. My sincere appreciation goes to my employer St John’s University of Tanzania for granting me study leave and financial support to pursue my PhD program. I also extend my sincere gratitude to Prof Dominic M. Kambarage and Prof Esron Karimuribo for the excellent financial support I received from the INTRA_ACP mobility project allowing me to carry out a part of the studies in Zambia. My heartfelt gratitude goes to Dr. Jennifer Stevenson for her tireless mentorship in Entomology and molecular assay during the entire period I visited Malaria Research Trust (MRT) at Macha-Choma Zambia. She was very instrumental in growth of my knowledge in Malaria Entomology. I extend my thankfulness to all MRT staff for their superb support in the laboratory. Special thanks go to Limonty Simubali, Ben Katowa, Jailos Lubinda, Natasha Laban and field and insectary team led by Bruice Malembeka, Alpha Simudoombe and Joe vii Mwaanga for their wonderful cooperation. My grateful thanks also go to Mr. Faraji Abilahi, Mr. Abbas K. Saidi, Mr. Francis A. Tumbo and other staff at Ifakara Health Institute (IHI) for their support in mosquito molecular and serological work done at their institute. Special gratitude goes to my colleagues Dr. Barnabas Mngumi, Dr. Joseph Mdachi, Mr Abdallah Husein and to the field team led by Mr. Hamisi Nyoni for giving me support in data collection and analysis. I am also thankful to all staff at SUA Pest Management Centre (SPMC) for excellent supervision when I was keeping mosquitoes in their insectary. All this could not be successful without Prof.Bukheti Kilonzo, Dr. Ladslaus L. Mnyone and Ms. Resta R. Maganga. Without commitments from the Administrative authorities of Morogoro and Dodoma municipalities and willingness of all study respondents, all activities related to the execution of this PhD programme would have been futile. I am deeply indebted to them for their excellent cooperation and willingness to participate and they were the main stakeholders for turning my PhD into reality. To my lovely friends Dr. Sabina P.Sheppa, Dr. Franscis Mdoe, Dr. David.Z Munisi, Dr.Tumbwene Mwansisya, Dr. Richard Mongi, Elisa Mwega, and Leonard Kamanga, I thank them all for keeping the boat afloat despite the massive turbulences in the rough seas. Last but not the least, I wish to express my sincere appreciation and heartfelt gratitude to my lovely husband Gibson Frank Mawalla and my sons Joshua, Jovin, Salema and my nieces Emiliana, Given, my sweet maids Fortunata and Diana for their moral support. I know they missed me when I was away but they always believed that “my success was also theirs”. Special thanks should also go to my lovely parents (Mr and Mrs Mathew Mathania), my beloved brothers and sisters as well as in-laws for their prayers and always giving me excellent moral support. “Thank you very much. May Almighty God bless you all.” viii DEDICATION This thesis is dedicated to my lovely husband, Gibson F. Mawalla, my adorable sons Joshua, Jovin and Salema, my niece Given and to my dear parents Mr. and Mrs. Mathew Mathania and beloved brothers and sisters. They have always offered encouragement and support in my endeavours. ix TABLE OF CONTENTS EXTENDED ABSTRACT .................................................................................................. ii DECLARATION ................................................................................................................ iv COPYRIGHT ....................................................................................................................... v ACKNOWLEDGEMENT .................................................................................................. vi DEDICATION .................................................................................................................. viii TABLE OF CONTENTS .................................................................................................... ix LIST OF TABLES ............................................................................................................ xiv LIST OF FIGURES ........................................................................................................
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    UNIVERSIDAD NACIONAL DE SAN AGUSTÍN DE AREQUIPA FACULTAD DE CIENCIAS BIOLÓGICAS ESCUELA PROFESIONAL DE BIOLOGÍA ASPECTOS ECOLÓGICOS Y BIOLÓGICOS DE LA MARIPOSA ENDÉMICA Mathania nov. sp. (LEPIDOPTERA: PIERIDAE) EN LOS ALREDEDORES DE AREQUIPA, 2014-2017. Tesis presentada por la Bachiller: CINTHIA MILAGROS BRAVO VELASQUEZ Para optar el Título Profesional de Biólogo. Asesor: Dr. EVARISTO LUCIANO LÓPEZ TEJEDA. AREQUIPA-PERÚ 2018 Dr. Evaristo Luciano López Tejeda Asesor Dr. Alberto Morales Hurtado Presidente Mg. Ana Lazo Rivera Secretaria Dr. Evaristo López Tejeda Miembro Dedico este trabajo a José, mi hermano, por enseñarme que, aun en las dificultades hay oportunidades para crecer, y que los límites son mentales. “Si tu intención es describir la verdad, hazlo con sencillez y la elegancia déjasela al sastre.” Albert Einstein. “No hay grandeza donde falte la sencillez, la bondad y la verdad” Lievi Tolstói. AGRADECIMIENTOS Los agradecimientos son infinitos; en primer Lugar a aquel ser divino que nunca me ha abandona, camina conmigo y me ha permitido llegar hasta este punto en esta vida, a Dios. A mi asesor el Dr. Evaristo López Tejeda, Director del Museo de Historia Natural de la Universidad Nacional de San Agustín (MUSA) y a mi co-asesor José Cerdeña, encargado del área de Entomología del MUSA, por todo el apoyo por los consejos y por su tiempo. A la Universidad Nacional de San Agustín por la subvención otorgada según el contrato N°019-2016-UNSA-INVESTIGA, permitieron la realización de esta Tesis. Mi agradecimiento especial para el Profesor Héctor Vargas, por su paciencia y buena disposición siempre para apoyarme, al compartir su vasta experiencia en el mundo de los “inmaduros”; y al profesor Hugo Benítez, por sus consejos al compartir conmigo sus experiencias, ambos de la Universidad de Tarapacá quienes fueron gran parte de la inspiración de seguir en el maravilloso mundo de la investigación.