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      The First Record of Echinococcus ortleppi (G5) Tapeworms in Grey Wolf ( Canis lupus)

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          Abstract

          The aim of this study is to confirm the presence and molecular identification of Echinococcus tapeworms in wolves from south-eastern Poland. An investigation was carried out on the intestines of 13 wolves from south-eastern Poland. The small intestines were divided into three equal segments. Each segment was separately examined using the sedimentation and counting technique (SCT). The detected Echinococcus tapeworms were isolated and identified by PCRs and sequencing ( nad1 and cox1 genes). Additionally, DNA isolated from the feces of wolves positive for Echinococcus tapeworms was examined with two diagnostic PCRs. The intestines of one wolf were positive for E. granulosus s.l. when assessed by SCT; the intestine was from a six-year-old male wolf killed in a communication accident. We detected 61 adult tapeworms: 42 in the anterior, 14 in the middle, and 5 in the posterior parts of the small intestine. The PCRs conducted for cox1 and nad1 produced specific products. A sequence comparison with the GenBank database showed similarity to the deposited E. ortleppi (G5) sequences. An analysis of the available phylogenetic sequences showed very little variation within the species of E. ortleppi (G5), and identity ranged from 99.10% to 100.00% in the case of cox1 and from 99.04% to 100.00% in the case of nad1. One of the two diagnostic PCRs used and performed on the feces of Echinococcus-positive animals showed product specific for E. granulosus. This study showed the presence of adult E. ortleppi tapeworms in wolves for the first time.

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          Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data

          Summary: The two main functions of bioinformatics are the organization and analysis of biological data using computational resources. Geneious Basic has been designed to be an easy-to-use and flexible desktop software application framework for the organization and analysis of biological data, with a focus on molecular sequences and related data types. It integrates numerous industry-standard discovery analysis tools, with interactive visualizations to generate publication-ready images. One key contribution to researchers in the life sciences is the Geneious public application programming interface (API) that affords the ability to leverage the existing framework of the Geneious Basic software platform for virtually unlimited extension and customization. The result is an increase in the speed and quality of development of computation tools for the life sciences, due to the functionality and graphical user interface available to the developer through the public API. Geneious Basic represents an ideal platform for the bioinformatics community to leverage existing components and to integrate their own specific requirements for the discovery, analysis and visualization of biological data. Availability and implementation: Binaries and public API freely available for download at http://www.geneious.com/basic, implemented in Java and supported on Linux, Apple OSX and MS Windows. The software is also available from the Bio-Linux package repository at http://nebc.nerc.ac.uk/news/geneiousonbl. Contact: peter@biomatters.com
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            Global Distribution of Alveolar and Cystic Echinococcosis.

            Alveolar echinococcosis (AE) and cystic echinococcosis (CE) are severe helminthic zoonoses. Echinococcus multilocularis (causative agent of AE) is widely distributed in the northern hemisphere where it is typically maintained in a wild animal cycle including canids as definitive hosts and rodents as intermediate hosts. The species Echinococcus granulosus, Echinococcus ortleppi, Echinococcus canadensis and Echinococcus intermedius are the causative agents of CE with a worldwide distribution and a highly variable human disease burden in the different endemic areas depending upon human behavioural risk factors, the diversity and ecology of animal host assemblages and the genetic diversity within Echinococcus species which differ in their zoonotic potential and pathogenicity. Both AE and CE are regarded as neglected zoonoses, with a higher overall burden of disease for CE due to its global distribution and high regional prevalence, but a higher pathogenicity and case fatality rate for AE, especially in Asia. Over the past two decades, numerous studies have addressed the epidemiology and distribution of these Echinococcus species worldwide, resulting in better-defined boundaries of the endemic areas. This chapter presents the global distribution of Echinococcus species and human AE and CE in maps and summarizes the global data on host assemblages, transmission, prevalence in animal definitive hosts, incidence in people and molecular epidemiology.
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              Identification of taeniid eggs in the faeces from carnivores based on multiplex PCR using targets in mitochondrial DNA.

              A multiplex polymerase chain reaction (PCR) was evaluated for the identification of morphologically indistinguishable eggs of the taeniid tapeworms from carnivores using primers targeting mitochondrial genes. The primers for Echinococcus multilocularis (amplicon size 395 bp) were species-specific as assessed by in silico analysis and in the PCR using well-defined control samples. The design of primers that specifically amplify DNA from E. granulosus or Taenia spp. was not possible. The primers designed for E. granulosus also amplified DNA (117 bp) from E. vogeli, and those designed for Taenia spp. amplified products (267 bp) from species of Mesocestoides, Dipylidium and Diphyllobothrium. Nevertheless, as our diagnostic approach includes the concentration of taeniid eggs by sequential sieving and flotation, followed by their morphological detection, this non-specificity has limited practical importance. Sequence analysis of the corresponding amplicon can identify most of the described E. granulosus genotypes. Taenia spp. can be identified by direct sequencing of the 267 bp amplicon, or, for most species, by restriction fragment length polymorphism (RFLP) analysis. The multiplex PCR was readily able to detect 1 egg (estimated to contain 7000 targets, as determined by quantitative PCR). Having been validated using a panel of well-defined samples from carnivores with known infection status, this approach proved to be useful for the identification of taeniid eggs from both individual animals and for epidemiological studies.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Pathogens
                Pathogens
                pathogens
                Pathogens
                MDPI
                2076-0817
                06 July 2021
                July 2021
                : 10
                : 7
                : 853
                Affiliations
                Author notes
                [* ]Correspondence: j.karamon@ 123456piwet.pulawy.pl ; Tel.: +48-81-889-3040
                Author information
                https://orcid.org/0000-0002-5830-167X
                https://orcid.org/0000-0003-2135-701X
                https://orcid.org/0000-0002-9982-3028
                https://orcid.org/0000-0002-7297-2082
                https://orcid.org/0000-0003-3649-7867
                Article
                pathogens-10-00853
                10.3390/pathogens10070853
                8308913
                34358003
                7be8bf38-9351-4826-8697-e537e36e150b
                © 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 ( https://creativecommons.org/licenses/by/4.0/).

                History
                : 02 June 2021
                : 28 June 2021
                Categories
                Article

                wolf,canis lupus,echinococcus granulosus,echinococcus ortleppi,poland

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