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      Multiple Tick-Borne Pathogens in Ixodes ricinus Ticks Collected from Humans in Romania

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          Abstract

          Ticks are medically important vectors of infectious diseases that are able to transmit pathogens to humans and animals. Tick-borne diseases represent a major health concern, posing an increasing risk to the public health during the last century and affecting millions of people. The aim of the current study was to provide epidemiological data regarding the presence of certain tick-borne pathogens in ticks feeding on humans in Romania. Overall, 522 Ixodes ricinus ticks collected from humans were screened for six pathogens: Borrelia spp., Neoehrlichia mikurensis, Babesia spp., Coxiella spp., Bartonella spp., and Francisella tularensis. Ticks attached to humans were collected between 2013–2015 in Cluj County, Romania. Conventional, nested and quantitative PCR were used to detect specific genetic sequences of each pathogen. For identifying the infectious agents, positive samples were sequenced. The infection prevalence was 21.07% from which 8.18% were mixed infections. The detected agents were Borrelia spp., N. mikurensis and Babesia spp. The present data reveal the endemic occurrence of potentially zoonotic pathogens in Romania. Revealing the current distribution of tick-borne pathogens in ticks collected from humans may provide new insights in understanding the complex ecology of tick-borne diseases and enlightens current knowledge about the infection prevalence at local, regional and national levels.

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          On a Cercopithifilaria sp. transmitted by Rhipicephalus sanguineus: a neglected, but widespread filarioid of dogs

          Background This study was aimed at investigating the distribution of a Cercopithifilaria sp. sensu Otranto et al., 2011 with dermal microfilariae recently identified in a dog from Sicily (Italy). A large epidemiological survey was conducted by examining skin samples (n = 917) and ticks (n = 890) collected from dogs at different time points in Italy, central Spain and eastern Greece. Results The overall prevalence of Cercopithifilaria sp. in the sampled animal populations was 13.9% and 10.5% by microscopy of skin sediments and by PCR on skin samples, respectively. Up to 21.6% and 45.5% of dogs in Spain were positive by microscopical examination and by PCR. Cumulative incidence rates ranging from 7.7% to 13.9% were estimated in dogs from two sites in Italy. A low level of agreement between the two diagnostic tests (microscopical examination and PCR) was recorded in sites where samples were processed in parallel. Infestation rate as determined by tick dissection (from 5.2% to 16.7%) was higher than that detected by PCR (from 0% to 3.9%); tick infestation was significantly associated with Cercopithifilaria sp. infestation in dogs from two out of four sites. Developing larvae found in ticks were morphometrically studied and as many as 1469 larvae were found in a single tick. Conclusions Our data suggest that, in addition to the most common species of filarioids known to infest dogs (i.e., Dirofilaria immitis, Dirofilaria repens and Acanthocheilonema reconditum), Cercopithifilaria sp. with dermal microfilariae should be considered due to its widespread distribution in southern Europe and high frequency in tick-exposed dogs.
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            Heat treatment prior to testing allows detection of antigen of Dirofilaria immitis in feline serum

            Background Diagnosis of Dirofilaria immitis infection in cats is complicated by the difficulty associated with reliable detection of antigen in feline blood and serum samples. Methods To determine if antigen-antibody complex formation may interfere with detection of antigen in feline samples, we evaluated the performance of four different commercially available heartworm tests using serum samples from six cats experimentally infected with D. immitis and confirmed to harbor a low number of adult worms (mean = 2.0). Sera collected 168 (n = 6), 196 (n = 6), and 224 (n = 6) days post infection were tested both directly and following heat treatment. Results Antigen was detected in serum samples from 0 or 1 of 6 infected cats using the assays according to manufacturer’s directions, but after heat treatment of serum samples, as many as 5 of 6 cats had detectable antigen 6–8 months post infection. Antibodies to D. immitis were detected in all six infected cats by commercial in-clinic assay and at a reference laboratory. Conclusions These results indicate that heat treatment of samples prior to testing can improve the sensitivity of antigen assays in feline patients, supporting more accurate diagnosis of this infection in cats. Surveys conducted by antigen testing without prior heat treatment of samples likely underestimate the true prevalence of infection in cats.
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              Infection prevalence and ecotypes of Anaplasma phagocytophilum in moose Alces alces , red deer Cervus elaphus , roe deer Capreolus capreolus and Ixodes ricinus ticks from Norway

              Background The geographical expansion of the tick Ixodes ricinus in northern Europe is a serious concern for animal and human health. The pathogen Anaplasma phagocytophilum is transmitted by ticks and causes emergences of tick-borne fever (anaplasmosis) in livestock. The transmission dynamics of the different ecotypes of A. phagocytophilum in the ecosystems is only partly determined. Red deer and roe deer contribute to circulation of different ecotypes of A. phagocytophilum in continental Europe, while the role of moose for circulation of different ecotypes is not fully established but an important issue in northern Europe. Methods We determined infection prevalence and ecotypes of A. phagocytophilum in moose (n = 111), red deer (n = 141), roe deer (n = 28) and questing ticks (n = 9241) in Norway. Results As previously described, red deer was exclusively linked to circulation of ecotype I, while roe deer was exclusively linked to circulation of ecotype II. Surprisingly, we found 58% ecotype I (n = 19) and 42% of ecotype II (n = 14) in moose. Both ecotypes were found in questing ticks in areas with multiple cervid species present, while only ecotype I was found in ticks in a region with only red deer present. Hence, the geographical distribution of ecotypes in ticks followed the distribution of cervid species present in a given region and their link to ecotype I and II. Conclusions Moose probably function as reservoirs for both ecotype I and II, indicating that the ecotypes of A. phagocytophilum are not entirely host-specific and have overlapping niches. The disease hazard depends also on both host abundance and the number of immature ticks fed by each host. Our study provides novel insights in the northern distribution and expansion of tick-borne fever.
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                Author and article information

                Journal
                Pathogens
                Pathogens
                pathogens
                Pathogens
                MDPI
                2076-0817
                19 May 2020
                May 2020
                : 9
                : 5
                : 390
                Affiliations
                [1 ]Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, 400372 Cluj-Napoca, Romania; mirabela.dumitrache@ 123456usamvcluj.ro (M.O.D.); gianluca.damico@ 123456usamvcluj.ro (G.D.); matei.ioana@ 123456usamvcluj.ro (I.A.M.); ionica.angela@ 123456usamvcluj.ro (A.M.I.); calin.gherman@ 123456usamvcluj.ro (C.M.G.); amihalca@ 123456usamvcluj.ro (A.D.M.)
                [2 ]Department of Microbiology, Immunology and Epidemiology, Faculty of Veterinary Medicine, Calea Mănăştur, no. 3–5, 400372 Cluj-Napoca, Romania
                [3 ]“Regele Mihai I al României” Life Sciences Institute, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, 3–5 Calea Mănăștur, 400372 Cluj-Napoca, Romania
                [4 ]Department of Infectious Diseases, University of Medicine and Pharmacy “Iuliu Haţieganu”, Iuliu Moldovan 23, 400000 Cluj-Napoca, Romania; mihaela.lupse@ 123456yahoo.com
                Author notes
                [* ]Correspondence: zsuzsa.kalmar@ 123456usamvcluj.ro ; Tel.: +4072-2834-993
                Author information
                https://orcid.org/0000-0001-7495-1392
                https://orcid.org/0000-0002-5936-2538
                Article
                pathogens-09-00390
                10.3390/pathogens9050390
                7281082
                32438768
                631d527d-c2b4-4992-96b7-195ee8376e46
                © 2020 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
                : 20 April 2020
                : 18 May 2020
                Categories
                Article

                ixodes ricinus,ticks,pathogens,tick-borne disease,humans
                ixodes ricinus, ticks, pathogens, tick-borne disease, humans

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