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      IDENTIFICATION OF Pseudomonas spp. AS AMOEBA-RESISTANT MICROORGANISMS IN ISOLATES OF Acanthamoeba Translated title: Identificação de Pseudomonas spp. como microrganismo resistente a ameba em isolados de Acanthamoeba

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          Abstract

          Acanthamoeba is a “Trojan horse” of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas.

          Translated abstract

          Acanthamoeba é um “Cavalo de Tróia” do mundo microbiano. Este estudo teve como objetivo identificar a presença de Pseudomonas como microrganismo resistente a ameba em 12 isolados de Acanthamoeba. Todos os isolados apresentaram o gênero Pseudomonas spp. como um microrganismo resistente a ameba. Assim, podemos ver que os isolados de Acanthamoeba estudados são hospedeiros de Pseudomonas.

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          Most cited references32

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          16S ribosomal DNA amplification for phylogenetic study.

          A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described along with methods for their use and examples. One pair of primers is capable of amplifying nearly full-length 16S ribosomal DNA (rDNA) from many bacterial genera; the additional primers are useful for various exceptional sequences. Methods for purification of amplified material, direct sequencing, cloning, sequencing, and transcription are outlined. An obligate intracellular parasite of bovine erythrocytes, Anaplasma marginale, is used as an example; its 16S rDNA was amplified, cloned, sequenced, and phylogenetically placed. Anaplasmas are related to the genera Rickettsia and Ehrlichia. In addition, 16S rDNAs from several species were readily amplified from material found in lyophilized ampoules from the American Type Culture Collection. By use of this method, the phylogenetic study of extremely fastidious or highly pathogenic bacterial species can be carried out without the need to culture them. In theory, any gene segment for which polymerase chain reaction primer design is possible can be derived from a readily obtainable lyophilized bacterial culture.
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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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              Microorganisms resistant to free-living amoebae.

              Free-living amoebae feed on bacteria, fungi, and algae. However, some microorganisms have evolved to become resistant to these protists. These amoeba-resistant microorganisms include established pathogens, such as Cryptococcus neoformans, Legionella spp., Chlamydophila pneumoniae, Mycobacterium avium, Listeria monocytogenes, Pseudomonas aeruginosa, and Francisella tularensis, and emerging pathogens, such as Bosea spp., Simkania negevensis, Parachlamydia acanthamoebae, and Legionella-like amoebal pathogens. Some of these amoeba-resistant bacteria (ARB) are lytic for their amoebal host, while others are considered endosymbionts, since a stable host-parasite ratio is maintained. Free-living amoebae represent an important reservoir of ARB and may, while encysted, protect the internalized bacteria from chlorine and other biocides. Free-living amoebae may act as a Trojan horse, bringing hidden ARB within the human "Troy," and may produce vesicles filled with ARB, increasing their transmission potential. Free-living amoebae may also play a role in the selection of virulence traits and in adaptation to survival in macrophages. Thus, intra-amoebal growth was found to enhance virulence, and similar mechanisms seem to be implicated in the survival of ARB in response to both amoebae and macrophages. Moreover, free-living amoebae represent a useful tool for the culture of some intracellular bacteria and new bacterial species that might be potential emerging pathogens.
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                Author and article information

                Journal
                Rev Inst Med Trop Sao Paulo
                Rev. Inst. Med. Trop. Sao Paulo
                Revista do Instituto de Medicina Tropical de São Paulo
                Instituto de Medicina Tropical
                0036-4665
                1678-9946
                Jan-Feb 2015
                Jan-Feb 2015
                : 57
                : 1
                : 81-83
                Affiliations
                Departamento de Microbiologia, Imunologia e Parasitologia. Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
                Author notes
                Correspondence to: Marilise Brittes Rott. Fax: +55 51 3308 3445. E-mail: marilise.rott@ 123456ufrgs.br
                Article
                10.1590/S0036-46652015000100012
                4325528
                25651331
                23b92021-9598-46b9-91a4-a0665bc1f703

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 20 September 2013
                : 28 March 2014
                Page count
                Figures: 1, Tables: 1, References: 17, Pages: 3
                Categories
                Brief Communication

                acanthamoeba,pseudomonas,amoeba-resistant microorganism

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