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      Exploring Prokaryotic and Eukaryotic Microbiomes Helps in Detecting Tick-Borne Infectious Agents in the Blood of Camels

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          Abstract

          Dromedary camels ( Camelus dromedarius) are widely distributed in Africa, the Middle East and northern India. In this study, we aimed to detect tick-borne pathogens through investigating prokaryotic and eukaryotic microorganisms in camel blood based on a metagenomic approach and then to characterize potentially pathogenic organisms using traditional molecular techniques. We showed that the bacteria circulating in the blood of camels is dominated by Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria. At the genus level, Sediminibacterium, Hydrotalea, Bradyrhizobium and Anaplasma were the most abundant taxa. Eukaryotic profile was dominated by Fungi, Charophyta and Apicomplexa. At the genus level, Theileria was detected in 10 out of 18 samples, while Sarcocystis, Hoplorhynchus and Stylocephalus were detected in one sample each. Our metagenomic approach was successful in the detection of several pathogens or potential pathogens including Anaplasma sp., Theileria ovis, Th. separata, Th. annulate, Th. mutans-like and uncharacterized Theileria sp. For further characterization, we provided the partial sequences of citrate synthase ( gltA) and heat-shock protein ( groEL) genes of Candidatus Anaplasma camelii. We also detected Trypanosoma evansi type A using polymerase chain reaction (PCR) targeting the internal transcribed spacer 1 (ITS1) region. This combined metagenomic and traditional approach will contribute to a better understanding of the epidemiology of pathogens including tick-borne bacteria and protozoa in animals.

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          MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability

          We report a major update of the MAFFT multiple sequence alignment program. This version has several new features, including options for adding unaligned sequences into an existing alignment, adjustment of direction in nucleotide alignment, constrained alignment and parallel processing, which were implemented after the previous major update. This report shows actual examples to explain how these features work, alone and in combination. Some examples incorrectly aligned by MAFFT are also shown to clarify its limitations. We discuss how to avoid misalignments, and our ongoing efforts to overcome such limitations.
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            MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms.

            The Molecular Evolutionary Genetics Analysis (Mega) software implements many analytical methods and tools for phylogenomics and phylomedicine. Here, we report a transformation of Mega to enable cross-platform use on Microsoft Windows and Linux operating systems. Mega X does not require virtualization or emulation software and provides a uniform user experience across platforms. Mega X has additionally been upgraded to use multiple computing cores for many molecular evolutionary analyses. Mega X is available in two interfaces (graphical and command line) and can be downloaded from www.megasoftware.net free of charge.
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              DADA2: High resolution sample inference from Illumina amplicon data

              We present DADA2, a software package that models and corrects Illumina-sequenced amplicon errors. DADA2 infers sample sequences exactly, without coarse-graining into OTUs, and resolves differences of as little as one nucleotide. In several mock communities DADA2 identified more real variants and output fewer spurious sequences than other methods. We applied DADA2 to vaginal samples from a cohort of pregnant women, revealing a diversity of previously undetected Lactobacillus crispatus variants.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Pathogens
                Pathogens
                pathogens
                Pathogens
                MDPI
                2076-0817
                16 March 2021
                March 2021
                : 10
                : 3
                : 351
                Affiliations
                [1 ]Laboratory of Parasitology, Faculty of Veterinary Medicine, Graduate School of Infectious Diseases, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan; wessam@ 123456czc.hokudai.ac.jp (W.M.A.M.); chatanga@ 123456vetmed.hokudai.ac.jp (E.C.); abassdoaa@ 123456yahoo.com (D.N.); nnonaka@ 123456vetmed.hokudai.ac.jp (N.N.)
                [2 ]Division of Infectious Diseases, Animal Medicine Department, Faculty of Veterinary Medicine, South Valley University, Qena 83523, Egypt; alsagher.ali@ 123456vet.svu.edu.eg (A.O.A.); mhassan@ 123456vet.svu.edu.eg (H.Y.A.H.M.)
                [3 ]Department of Parasitology, Faculty of Veterinary Medicine, South Valley University, Qena 83523, Egypt; dr_mosab2081@ 123456yahoo.com
                [4 ]Department of Veterinary Medicine, College of Agriculture and Veterinary Medicine, Qassim University, Buraidah 51452, Saudi Arabia
                [5 ]Department of Veterinary Medicine, Lilongwe University of Agriculture and Natural Resources, Lilongwe P.O. BOX 219, Malawi
                [6 ]Department of Parasitology, Faculty of Veterinary Medicine, University of Khartoum, Khartoum North 11111, Sudan; bashirsalim@ 123456gmail.com
                [7 ]Department of Hygiene and Zoonoses, Mansoura University, Mansoura 35516, Egypt
                [8 ]Graduate School of Environmental Science, Hokkaido University, Sapporo, Hokkaido 060-0810, Japan; janderslee@ 123456ees.hokudai.ac.jp
                [9 ]Division of Wildlife Medicine, Animal Medicine Department, Faculty of Veterinary Medicine, South Valley University, Qena 83523, Egypt
                Author notes
                [* ]Correspondence: m.abdallah@ 123456vetmed.hokudai.ac.jp (M.A.M.M.); ryo.nakao@ 123456vetmed.hokudai.ac.jp (R.N.); Tel./Fax: +81-11-706-5196 (M.A.M.M.); Tel./Fax: +81-11-706-5196 (R.N.)
                Author information
                https://orcid.org/0000-0002-9088-9207
                https://orcid.org/0000-0002-2301-0258
                https://orcid.org/0000-0003-1553-1763
                https://orcid.org/0000-0001-5448-2605
                https://orcid.org/0000-0002-3665-2217
                https://orcid.org/0000-0002-9413-694X
                https://orcid.org/0000-0002-3105-7603
                Article
                pathogens-10-00351
                10.3390/pathogens10030351
                8002256
                33809738
                1eedd1be-19af-4d5f-8b82-3dee6c0508c5
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 22 January 2021
                : 09 March 2021
                Categories
                Article

                candidatus anaplasma camelii,eukaryotes,microbiome,theileria,trypanosoma evansi

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