University of Texas at El Paso DigitalCommons@UTEP Open Access Theses & Dissertations 2015-01-01 Identification Of Leishmania Spp. And T. Cruzi Parasites In Stray Felines In El Paso, Texas Through Polymerase Chain Reaction Patricia Isabela Gonzalez University of Texas at El Paso, [email protected] Follow this and additional works at: https://digitalcommons.utep.edu/open_etd Part of the Public Health Education and Promotion Commons Recommended Citation Gonzalez, Patricia Isabela, "Identification Of Leishmania Spp. And T. Cruzi Parasites In Stray Felines In El Paso, Texas Through Polymerase Chain Reaction" (2015). Open Access Theses & Dissertations. 851. https://digitalcommons.utep.edu/open_etd/851 This is brought to you for free and open access by DigitalCommons@UTEP. It has been accepted for inclusion in Open Access Theses & Dissertations by an authorized administrator of DigitalCommons@UTEP. For more information, please contact [email protected]. IDENTIFICATION OF LEISHMANIA SPP. AND T. CRUZI PARASITES IN STRAY FELINES IN EL PASO, TEXAS THROUGH POLYMERASE CHAIN REACTION PATRICIA ISABELA GONZALEZ Master’s Program in Public Health APPROVED: Gabriel Ibarra-Mejia, M.D., MSErg, Ph.D., Chair Delfina Dominguez, Ph.D. Ken Waldrup, DVM, Ph.D. Rodrigo X. Armijos M.D., ScD, MSc Charles Ambler, Ph.D. Dean of the Graduate School Copyright © by Patricia Isabela Gonzalez 2015 IDENTIFICATION OF LEISHMANIA SPP. AND T. CRUZI PARASITES IN STRAY FELINES IN EL PASO, TEXAS THROUGH POLYMERASE CHAIN REACTION by PATRICIA ISABELA GONZALEZ, B.S. THESIS PROPOSAL Presented to the Faculty of the Graduate School of The University of Texas at El Paso in Partial Fulfillment of the Requirements for the Degree of MASTER OF PUBLIC HEALTH Department of Public Health Sciences THE UNIVERSITY OF TEXAS AT EL PASO December 2015 ACKNOWLEDGEMENTS This material is based on work supported by Department of Public Health Sciences from the University of Texas at El Paso. I thank specifically Dr. Gabriel Ibarra-Mejia, Dr. Delfina Dominguez, and Dr. Rodrigo Armijos that critically facilitated this work, Dr. Ken Waldrup and the El Paso Animal Services for granting permission and training for the collection of sample, and the Molecular Core Facility at the Border Biomedical Research Center (BBRC) for providing DNA sequencing service. I thank Dr. Maria Duarte, Chair of Department of Public Health Sciences at the University of Texas at El Paso, and Dr. Rodrigo Armijos for supporting this project by the Peter de Witter Endowed Chair funds. I would also like to thank Evan Kipp for assisting with the weekly collection of tissue samples from the City of El Paso Animal Control Facility and Emma Mejia and Cristina Martinez for assisting with the DNA extractions. iv ABSTRACT Chagas disease and leishmaniasis are considered neglected parasitic diseases, and although dogs are considered the main domestic reservoirs, infected cats have recently been found in endemic areas in several countries and became a public health concern. These diseases can be transmissible to other animals and are known to infect humans as well. Natural infection of cats by Leishmania spp. and Trypanosoma cruzi has been demonstrated in several European, Latin American, and Asian countries. A recent field study found dogs and wild animals tested positive for both Leishmania and Trypanosoma cruzi in the El Paso, Texas Region. Cats, however, have not been tested in this region. In this study we aimed to detect Leishmania parasites and T. cruzi in tissue samples from 155 stray cats from the El Paso, Texas Region. Samples from the spleen, skin, and heart of each cat were collected and subjected to molecular analysis (PCR). Positive samples where then sequenced as an alternate method to detect the presence of DNA from Leishmania parasites and T. cruzi. Percentage of frequency of positive samples was calculated and their distribution was mapped throughout El Paso region and surrounding areas according to place of capture. PCR results show positive identification of Leishmania spp. in nearly twenty percent of skin tissues, and no cases of T. cruzi in any of the tissues tested. DNA sequencing proved infection of Leishmania spp. parasites in 19 of the 155 cats indicative of the presence of the parasites in the tissues of the studied stray cat population. No geographical pattern was observed among the positive samples. v TABLE OF CONTENTS ACKNOWLEDGEMENTS ........................................................................................................... iv TABLE OF CONTENTS ............................................................................................................... vi LIST OF FIGURES ....................................................................................................................... ix LIST OF TABLES .......................................................................................................................... x INTRODUCTION .......................................................................................................................... 1 BACKGROUND AND SIGNIFICANCE ...................................................................................... 2 Leishmaniasis .............................................................................................................................. 2 Leishmaniasis Worldwide ....................................................................................................... 2 Leishmaniasis in Texas ........................................................................................................... 2 Leishmaniasis in El Paso ........................................................................................................ 3 Leishmaniasis in Cats ............................................................................................................. 3 Vector for Transmission of Leishmania ................................................................................. 4 Leishmania Life Cycle ............................................................................................................ 5 Leishmaniases Clinical Manifestations in Relation to the Species ......................................... 6 Chagas Disease ........................................................................................................................... 7 Chagas Disease and Trypanosoma Cruzi Worldwide ............................................................. 7 T. cruzi and Chagas Disease in Texas ..................................................................................... 8 T. cruzi in El Paso ................................................................................................................... 8 T. cruzi in Cats ........................................................................................................................ 9 Vector for Transmission of T. cruzi ........................................................................................ 9 T. cruzi Life Cycle ................................................................................................................ 11 vi Clinical Manifestations of Chagas Disease ........................................................................... 12 Environmental Factors .............................................................................................................. 13 Impact of Climate Change in the Spread of the Disease ...................................................... 13 Cats are Reservoirs in the Environment and Serve as Pets for Humans ............................... 13 Social and Behavioral Factors .................................................................................................. 14 Border Economic Changes in Relation to the Spread of the Disease ................................... 14 Methodology of Preference ........................................................................................................... 15 Polymerase Chain Reaction .................................................................................................. 15 Sequencing ............................................................................................................................ 15 Organs of Preference ................................................................................................................. 15 STUDY RATIONALE ................................................................................................................. 16 GOAL AND OBJECTIVES ......................................................................................................... 17 Problem Statement .................................................................................................................... 17 Research Question .................................................................................................................... 17 Overall Goal of the Study ......................................................................................................... 17 Objectives ................................................................................................................................. 17 METHODOLOGY ....................................................................................................................... 18 Experimental Design ................................................................................................................. 18 Sample Population ...................................................................................................................