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      Spotted Fever: Epidemiology and Vector- Rickettsia-Host Relationship in Rio de Janeiro State

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          Abstract

          The eco-epidemiological scenario of spotted fever (SF), a tick-borne disease that affects humans and other animals in several countries around the world, was analyzed in Rio de Janeiro (RJ) State, Brazil. During the last 34 years, 990 SF cases were reported in RJ (the Brazilian state with the highest population density), including 116 cases confirmed by serology (RIFI) or PCR, among 42.39% of the municipalities with reported cases of SF. The epidemiologic dynamics of SF in RJ State are very heterogeneous in time and space, with outbreaks, high mortality rates and periods of epidemiological silence (no SF cases reported). Furthermore, it exhibited a changing epidemiological profile from being rural to becoming an urban disease. This study identified arthropods infected with Rickettsia felis, R. bellii and R. rickettsii, and found that the abundance of ectoparasites was associated with specific hosts. The R. rickettsii-vector-host relationship was most evident in species-specific parasitism. This suggests that the association between dogs, cattle, horses, capybaras and their main ectoparasites, Rhipicephalus sanguineus and Ctenocephalides felis, Rhipicephalus microplus, Dermacentor nitens, and Amblyomma dubitatum, respectively, has a key role in the dynamics of R. rickettsii transmission in enzootic cycles and the maintenance of carrier ectoparasites, thus facilitating the existence of endemic areas with the ability to produce epidemic outbreaks of SF in RJ. This study found confirmed human infections for only the R. rickettsii carrier Amblyomma sculptum, which reinforces the importance of this species as a vector of the pathogen in Brazil. This study can be adapted to different eco-epidemiological scenarios of spotted fever throughout the Americas.

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          Molecular Cloning : A Laboratory Manual

          <p>The first two editions of this manual have been mainstays of molecular biology for nearly twenty years, with an unrivalled reputation for reliability, accuracy, and clarity.<br>In this new edition, authors Joseph Sambrook and David Russell have completely updated the book, revising every protocol and adding a mass of new material, to broaden its scope and maintain its unbeatable value for studies in genetics, molecular cell biology, developmental biology, microbiology, neuroscience, and immunology.<br>Handsomely redesigned and presented in new bindings of proven durability, this three–volume work is essential for everyone using today’s biomolecular techniques.<br>The opening chapters describe essential techniques, some well–established, some new, that are used every day in the best laboratories for isolating, analyzing and cloning DNA molecules, both large and small.<br>These are followed by chapters on cDNA cloning and exon trapping, amplification of DNA, generation and use of nucleic acid probes, mutagenesis, and DNA sequencing.<br>The concluding chapters deal with methods to screen expression libraries, express cloned genes in both prokaryotes and eukaryotic cells, analyze transcripts and proteins, and detect protein–protein interactions.<br>The Appendix is a compendium of reagents, vectors, media, technical suppliers, kits, electronic resources and other essential information.<br>As in earlier editions, this is the only manual that explains how to achieve success in cloning and provides a wealth of information about why techniques work, how they were first developed, and how they have evolved. </p>
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            Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques.

            A very simple, fast, universally applicable and reproducible method to extract high quality megabase genomic DNA from different organisms is described. We applied the same method to extract high quality complex genomic DNA from different tissues (wheat, barley, potato, beans, pear and almond leaves as well as fungi, insects and shrimps' fresh tissue) without any modification. The method does not require expensive and environmentally hazardous reagents and equipment. It can be performed even in low technology laboratories. The amount of tissue required by this method is approximately 50-100 mg. The quantity and the quality of the DNA extracted by this method is high enough to perform hundreds of PCR-based reactions and also to be used in other DNA manipulation techniques such as restriction digestion, Southern blot and cloning.
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              Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes.

              DNA sequences from specific genes, amplified by the polymerase chain reaction technique, were used as substrata for nonisotopic restriction endonuclease fragment length polymorphism differentiation of rickettsial species and genotypes. The products amplified using a single pair of oligonucleotide primers (derived from a rickettsial citrate synthase gene sequence) and cleaved with restriction endonucleases were used to differentiate almost all recognized species of rickettsiae. A second set of primers was used for differentiation of all recognized species of closely related spotted fever group rickettsiae. The procedure circumvents many technical obstacles previously associated with identification of rickettsial species. Multiple amplified DNA digest patterns were used to estimate the intraspecies nucleotide sequence divergence for the genes coding for rickettsial citrate synthase and a large antigen-coding gene of the spotted fever group rickettsiae. The estimated relationships deduced from these genotypic data correlate reasonably well with established rickettsial taxonomic schemes.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                30 March 2017
                2017
                : 8
                : 505
                Affiliations
                [1] 1Laboratório de Doenças Parasitária, Instituto Oswaldo Cruz/Fundação Oswaldo Cruz Rio de Janeiro, Brazil
                [2] 2Laboratório de Referência Nacional em Vetores das Riquetsioses – Secretaria de Vigilância em Saúde/Ministério da Saúde, Instituto Oswaldo Cruz/Fundação Oswaldo Cruz Rio de Janeiro, Brazil
                [3] 3Secretaria de Vigilância em Saúde – Ministério da Saúde Brasilia, Brazil
                [4] 4Secretaria de Estado de Saúde do Rio de Janeiro Rio de Janeiro, Brazil
                Author notes

                Edited by: Leonard Peruski, US Centers for Disease Control and Prevention, USA

                Reviewed by: Max Maurin, Université Grenoble Alpes, France; Karen A. Krogfelt, Statens Serum Institut, Denmark; Gerardo Acosta-Jamett, Austral University of Chile, Chile

                *Correspondence: Gilberto S. Gazêta, gsgazeta@ 123456ioc.fiocruz.br

                This article was submitted to Infectious Diseases, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2017.00505
                5371726
                4ec8a73f-9bf8-421b-bde9-8c63256cb0b5
                Copyright © 2017 Montenegro, Bitencourth, de Oliveira, Borsoi, Cardoso, Sousa, Giordano-Dias, Amorim, Serra-Freire, Gazêta and Brazil.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 03 May 2016
                : 10 March 2017
                Page count
                Figures: 5, Tables: 1, Equations: 0, References: 57, Pages: 10, Words: 0
                Funding
                Funded by: Coordenação de perfeiçoamento de Pessoal de Nível Superior 10.13039/501100002322
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                public health,eco-epidemiology,rickettsioses,tick-borne diseases,zoonosis
                Microbiology & Virology
                public health, eco-epidemiology, rickettsioses, tick-borne diseases, zoonosis

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