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      Infección con Toxoplasma gondii (Eucoccidiorida: Sarcocystidae) en murciélagos de Campeche y Yucatán, México Translated title: Infection with Toxoplasma gondii (Eucoccidiorida: Sarcocystidae) in bats of Campeche and Yucatán, Mexico

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          Abstract

          Resumen Toxoplasma gondii es un protozoario parásito reconocido como el agente causal de la toxoplasmosis, enfermedad zoonótica que afecta a humanos y animales domésticos o silvestres. En México, representa un problema de salud pública y veterinaria, sobre todo en regiones con climas tropicales y subtropicales. Los murciélagos han sido identificados como hospederos accidentales en el ciclo de transmisión; no obstante, en México no existe información previa; por lo tanto, el objetivo del presente estudio es reportar la infección con T. gondii en murciélagos capturados en sitios de los estados de Campeche y Yucatán, México. Se capturaron murciélagos en dos sitios de Yucatán y uno de Campeche, ubicados en la Península de Yucatán. Se recolectaron riñones, bazo e hígado y se emplearon en la extracción de ADN total. La infección con T. gondii se detectó a través de la amplificación de un fragmento del gen B1, utilizando PCR anidada. Los productos positivos fueron purificados y enviados a secuenciación para su posterior análisis de alineamiento; adicionalmente, se construyó un árbol filogenético. Se analizaron un total de 69 murciélagos pertenecientes a ocho especies distintas: 41 (59.4 %, 41/69) Artibeus jamaicensis; seis (8.7 %, 6/69) Pteronotus parnellii; seis (8.7 %, 6/69) Noctilio leporinus; seis (8.7 %, 6/69) Chiroderma villosum; cuatro (5.8 %, 4/69) Glossophaga soricina; dos (2.9 %, 2/69) Carollia sowelli; dos (2.89 %, 2/69) Artibeus lituratus y dos (2.9%, 2/69) Rhogeessa aeneus. La PCR anidada identificó ocho (11.6 %, 8/69) murciélagos positivos a la infección: seis (75 %, 6/8) A. jamaicensis, capturados en X’matkuil y Panabá, un (12.5 %, 1/8) G. soricina y un (12.5 %, 1/8) C. villosum, ambos capturados en Panabá. El análisis de alineamiento arrojó 99-100 % para cobertura y 97-99 % para identidad respecto a secuencias de T. gondii. Nuestros resultados aportan al entendimiento del ciclo de transmisión de T. gondii en la región; sin embargo, son necesarias investigaciones futuras para determinar los genotipos circulantes, ya que estudios anteriores han demostrado que estos animales pueden estar infectados con genotipos identificados en otros animales domésticos o silvestres e incluso en humanos.

          Translated abstract

          Abstract Toxoplasma gondii is a protozoan parasite recognized as the causative agent of toxoplasmosis, a zoonotic disease that affects humans and domestic or wild animals. In Mexico, it represents a public and animal health problem, especially in regions with tropical and subtropical climates. Bats have been reported as accidental hosts in the transmission cycle; however, there is no preceding information in Mexico. Therefore, the aim of the present study is to report the T. gondii infection in bats captured in sites of Campeche and Yucatan states, Mexico. Bats were captured in two sites in Yucatan (X’matkuil and Panaba) and one in Campeche (Hampolol), located in the Yucatan Peninsula. Kidneys, spleen, and liver were collected and used in the total DNA extraction. Toxoplasma gondii infection was detected through the amplification of a B1 gene fragment, using nested PCR. The positive PCR products were purified and sent to sequencing for a posterior sequence identity analysis. Additionally, a phylogenetic tree was made. A total of 69 bats belonging to eight different species were processed: 41 (59.4 %, 41/69) Artibeus jamaicensis; six (8.7 %, 6/69) Pteronotus parnellii; six (8.7 %, 6/69) Noctilio leporinus; six (8.7 %, 6/69) Chiroderma villosum; four (5.8 %, 4/69) Glossophaga soricina; two (2.9 %, 2/69) Carollia sowelli; two (2.89 %, 2/69) Artibeus lituratus; and two (2.9 %, 2/69) Rhogeessa aeneus. The nested PCR identified eight (11.6 %, 8/69) infected bats: six (75 %, 6/8) A. jamaicensis, captured in X'matkuil and Panaba, one (12.5 %, 1/8) G. soricina, and one (12.5 %, 1/8) C. villosum, both captured in Panaba. The alignment analysis yielded 99-100 % for cover and 97-99 % for identity to T. gondii sequences. Our results contribute to the understanding of the T. gondii transmission cycle in the region; however, future research is needed to determine circulating genotypes, as previous studies have demonstrated that these animals might be infected with identified genotypes in other domestic or wild animals and even in humans.

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          MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets.

          We present the latest version of the Molecular Evolutionary Genetics Analysis (Mega) software, which contains many sophisticated methods and tools for phylogenomics and phylomedicine. In this major upgrade, Mega has been optimized for use on 64-bit computing systems for analyzing larger datasets. Researchers can now explore and analyze tens of thousands of sequences in Mega The new version also provides an advanced wizard for building timetrees and includes a new functionality to automatically predict gene duplication events in gene family trees. The 64-bit Mega is made available in two interfaces: graphical and command line. The graphical user interface (GUI) is a native Microsoft Windows application that can also be used on Mac OS X. The command line Mega is available as native applications for Windows, Linux, and Mac OS X. They are intended for use in high-throughput and scripted analysis. Both versions are available from www.megasoftware.net free of charge.
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            A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences.

            Some simple formulae were obtained which enable us to estimate evolutionary distances in terms of the number of nucleotide substitutions (and, also, the evolutionary rates when the divergence times are known). In comparing a pair of nucleotide sequences, we distinguish two types of differences; if homologous sites are occupied by different nucleotide bases but both are purines or both pyrimidines, the difference is called type I (or "transition" type), while, if one of the two is a purine and the other is a pyrimidine, the difference is called type II (or "transversion" type). Letting P and Q be respectively the fractions of nucleotide sites showing type I and type II differences between two sequences compared, then the evolutionary distance per site is K = -(1/2) ln [(1-2P-Q) square root of 1-2Q]. The evolutionary rate per year is then given by k = K/(2T), where T is the time since the divergence of the two sequences. If only the third codon positions are compared, the synonymous component of the evolutionary base substitutions per site is estimated by K'S = -(1/2) ln (1-2P-Q). Also, formulae for standard errors were obtained. Some examples were worked out using reported globin sequences to show that synonymous substitutions occur at much higher rates than amino acid-altering substitutions in evolution.
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              Confidence Limits on Phylogenies: An Approach Using the Bootstrap

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                Author and article information

                Journal
                rbt
                Revista de Biología Tropical
                Rev. biol. trop
                Universidad de Costa Rica (San José, San José, Costa Rica )
                0034-7744
                0034-7744
                June 2019
                : 67
                : 3
                : 633-642
                Affiliations
                [1] Yucatán orgnameUniversidad Autónoma de Yucatán Mexico antonio.torres@ 123456correo.uady.mx
                [5] Yucatán orgnameLaboratorio DIMyGEN México javiersosae@ 123456hotmail.com
                [4] Antioquía orgnameUniversidad CES Colombia rpelaezp@ 123456ces.edu.co
                [2] Yucatán orgnameUniversidad Autónoma de Yucatán Mexico hbetanc@ 123456correo.uady.mx
                [3] Yucatán orgnameUniversidad Autónoma de Yucatán Mexico bgonza@ 123456correo.uady.mx
                Article
                S0034-77442019000300633 S0034-7744(19)06700300633
                10.15517/rbt.v67i3.35147
                f5c1200f-5335-4deb-928e-82ce1d9fb9c2

                This work is licensed under a Creative Commons Attribution 3.0 International License.

                History
                : 07 May 2019
                : 12 November 2018
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 40, Pages: 10
                Product

                SciELO Costa Rica

                Categories
                Artículo

                Toxoplasma gondii,PCR anidada,B1,infección,quirópteros,ciclo de transmisión,nested PCR,infection,Chiroptera,transmission cycle

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