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      Molecular Characterization of Lipoptena fortisetosa from Environmental Samples Collected in North-Eastern Poland

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          Abstract

          Simple Summary

          Lipoptena fortisetosa is an invasive, hematophagous insect, which lives on cervids and continues to spread across Europe. The species originated from the Far East and eastern Siberia. Besides wild animals, these ectoparasites can attack humans, companion animals, and livestock. These insects may also play a role in transmitting infectious diseases. The objective of this study was to confirm the presence of L. fortisetosa in north-eastern Poland and to characterize the examined population with the use of molecular methods. Deer keds were collected from six natural forests in the region of Warmia and Mazury. DNA of L. fortisetosa was extracted and subjected to molecular studies. Two species of deer keds ( Lipoptena cervi and L. fortisetosa) were obtained in each location during field research. There were no differences in the sex distribution of these two ectoparasite species. During the research, more L. cervi than L. fortisetosa specimens were obtained. The studied insects were very closely related to specimens from Lithuania, the Czech Republic, and Japan. Our study indicates various ectoparasite lineages, and such research contributes to the improvement of basic knowledge on the distribution, genetic structure, and variability of the invasive ked fly L. fortisetosa.

          Abstract

          Recent years have witnessed an increase in the population of Lipoptena fortisetosa in Central Europe. The genetic profile of this ectoparasite has not been studied in Poland to date. The aim of the present study was to confirm the presence of L. fortisetosa in north-eastern Poland and to characterize the examined population with the use of molecular methods. Deer keds were collected between June and July 2019 in six natural, mixed forests. A fragment of the rRNA 16S gene was used as a marker to identify L. fortisetosa by polymerase chain reaction (PCR). DNA samples were sequenced in the last step. Six new locations of L. fortisetosa were confirmed. No significant differences were observed in the sex ratios of L. cervi and L. fortisetosa ( L. cervi p-value = 0.74; L. fortisetosa p-value = 0.65). Significant differences were noted between the total size of L. cervi and L. fortisetosa populations ( p-value < 0.001). The similarity to GenBank sequences ranged from 95.56% to 100%. The obtained nucleotide sequences were very closely related to L. fortisetosa sequences from Lithuania, the Czech Republic, and Japan. Molecular analyses revealed considerable genetic diversity, which could indicate that various ectoparasite lineages have spread throughout Europe.

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          BioEdit - a user friendly biological sequence alignment editor and analysis program for Windows 95/98/NT

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            Comparison of flagging, walking, trapping, and collecting from hosts as sampling methods for northern deer ticks, Ixodes dammini, and lone-star ticks, Amblyomma americanum (Acari:Ixodidae).

            Ticks were sampled by flagging, collecting from the investigator's clothing (walking samples), trapping with dry-ice bait, and collecting from mammal hosts on Fire Island, NY, U.S.A. The habitat distribution of adult deer ticks, Ixodes dammini, was the same in simultaneous collections from the investigator's clothing and from muslin flags. Walking and flagging samples can both be biased by differences between investigators, so the same person should do comparative samples whenever possible. Walking samples probably give a more accurate estimate than flagging samples of the human risk of encountering ticks. However, ticks (such as immature I. dammini) that seek hosts in leaf litter and ground-level vegetation are poorly sampled by walking collections. These ticks can be sampled by flagging at ground level. Dry-ice-baited tick-traps caught far more lone-star ticks, Amblyomma americanum, than deer ticks, even in areas where deer ticks predominated in flagging samples. In comparisons of tick mobility in the lab, nymphal A. americanum were more mobile than nymphal I. dammini in 84% of the trials. Therefore, the trapping bias may result from increased trap encounter due to more rapid movement by A. americanum, although greater attraction to carbon dioxide may also play a role. Tick traps are useful for intraspecific between-habitat comparisons. Early in their seasonal activity period, larval I. dammini were better represented in collections from mouse hosts than in flagging samples. Apparently, sampling from favored hosts can detect ticks at low population levels, but often cannot be used to get accurate estimates of pathogen prevalence in questing ticks.
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              The phylogeny and evolution of host choice in the Hippoboscoidea (Diptera) as reconstructed using four molecular markers.

              Hippoboscoidea is a superfamily of Diptera that contains the Glossinidae or tsetse flies, the Hippoboscidae or louse flies, and two families of bat flies, the Streblidae and the Nycteribiidae. We reconstruct the phylogenetic relationships within Hippoboscoidea using maximum parsimony and Bayesian methods based on nucleotide sequences from fragments of four genes: nuclear 28S ribosomal DNA and the CPSase domain of CAD, and mitochondrial 16S rDNA and cytochrome oxidase I. We recover monophyly for most of the presently recognized groups within Hippoboscoidea including the superfamily as a whole, the Hippoboscidae, the Nycteribiidae, the bat flies, and the Pupipara (=Hippoboscidae+Nycteribiidae+Streblidae), as well as several subfamilies within the constituent families. Streblidae appear to be paraphyletic. Our phylogenetic hypothesis is well supported and decisive in that most competing topological hypotheses for the Hippoboscoidea require significantly longer trees. We confirm a single shift from a free-living fly to a blood-feeding ectoparasite of vertebrates and demonstrate that at least two host shifts from mammals to birds have occurred. Wings have been repeatedly lost, but never regained. The hippoboscoid ancestor also evolved adenotrophic viviparity and our cladogram is consistent with a gradual reduction in the motility of the deposited final instar larvae from active burrowing in the soil to true pupiparity where adult females glue the puparium within the confines of bat roosts.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Animals (Basel)
                Animals (Basel)
                animals
                Animals : an Open Access Journal from MDPI
                MDPI
                2076-2615
                12 April 2021
                April 2021
                : 11
                : 4
                : 1093
                Affiliations
                [1 ]Department of Veterinary Prevention and Feed Hygiene, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, Poland; bakta@ 123456uwm.edu.pl
                [2 ]National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan; gen@ 123456obihiro.ac.jp
                [3 ]Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, Poland; jerzyj@ 123456uwm.edu.pl
                Author notes
                Author information
                https://orcid.org/0000-0002-1005-3945
                https://orcid.org/0000-0003-2780-110X
                https://orcid.org/0000-0001-6838-2840
                Article
                animals-11-01093
                10.3390/ani11041093
                8069113
                0503f167-c460-44a7-a491-2054fcee891c
                © 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
                : 24 March 2021
                : 08 April 2021
                Categories
                Article

                deer keds,ectoparasite,hippoboscidae,pcr,phylogenetics,vector
                deer keds, ectoparasite, hippoboscidae, pcr, phylogenetics, vector

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