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      Molecular detection of pathogens in ticks and fleas collected from companion dogs and cats in East and Southeast Asia

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          Abstract

          Background

          Ticks and fleas are considered amongst the most important arthropod vectors of medical and veterinary concern due to their ability to transmit pathogens to a range of animal species including dogs, cats and humans. By sharing a common environment with humans, companion animal-associated parasitic arthropods may potentially transmit zoonotic vector-borne pathogens (VBPs). This study aimed to molecularly detect pathogens from ticks and fleas from companion dogs and cats in East and Southeast Asia.

          Methods

          A total of 392 ticks and 248 fleas were collected from 401 infested animals (i.e. 271 dogs and 130 cats) from China, Taiwan, Indonesia, Malaysia, Singapore, Thailand, the Philippines and Vietnam, and molecularly screened for the presence of pathogens. Ticks were tested for Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. while fleas were screened for the presence of Rickettsia spp. and Bartonella spp.

          Result

          Of the 392 ticks tested, 37 (9.4%) scored positive for at least one pathogen with Hepatozoon canis being the most prevalent (5.4%), followed by Ehrlichia canis (1.8%), Babesia vogeli (1%), Anaplasma platys (0.8%) and Rickettsia spp. (1%) [including Rickettsia sp. (0.5%), Rickettsia asembonensis (0.3%) and Rickettsia felis (0.3%)]. Out of 248 fleas tested, 106 (42.7%) were harboring at least one pathogen with R. felis being the most common (19.4%), followed by Bartonella spp. (16.5%), Rickettsia asembonensis (10.9%) and “ Candidatus Rickettsia senegalensis” (0.4%). Furthermore, 35 Rhipicephalus sanguineus ticks were subjected to phylogenetic analysis, of which 34 ticks belonged to the tropical and only one belonged to the temperate lineage ( Rh. sanguineus ( sensu stricto)).

          Conclusion

          Our data reveals the circulation of different VBPs in ticks and fleas of dogs and cats from Asia, including zoonotic agents, which may represent a potential risk to animal and human health.

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          Most cited references77

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          Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes.

          DNA sequences from specific genes, amplified by the polymerase chain reaction technique, were used as substrata for nonisotopic restriction endonuclease fragment length polymorphism differentiation of rickettsial species and genotypes. The products amplified using a single pair of oligonucleotide primers (derived from a rickettsial citrate synthase gene sequence) and cleaved with restriction endonucleases were used to differentiate almost all recognized species of rickettsiae. A second set of primers was used for differentiation of all recognized species of closely related spotted fever group rickettsiae. The procedure circumvents many technical obstacles previously associated with identification of rickettsial species. Multiple amplified DNA digest patterns were used to estimate the intraspecies nucleotide sequence divergence for the genes coding for rickettsial citrate synthase and a large antigen-coding gene of the spotted fever group rickettsiae. The estimated relationships deduced from these genotypic data correlate reasonably well with established rickettsial taxonomic schemes.
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            Experimental transmission of Bartonella henselae by the cat flea.

            Bartonella henselae is an emerging bacterial pathogen, causing cat scratch disease and bacillary angiomatosis. Cats bacteremic with B. henselae constitute a large reservoir from which humans become infected. Prevention of human infection depends on elucidation of the natural history and means of feline infection. We studied 47 cattery cats in a private home for 12 months to determine the longitudinal prevalence of B. henselae bacteremia, the prevalence of B. henselae in the fleas infesting these cats, and whether B. henselae is transmitted experimentally to cats via fleas. Vector-mediated transmission of B.henselae isolates was evaluated by removing fleas from the naturally bacteremic, flea-infested cattery cats and transferring these fleas to specific-pathogen-free (SPF) kittens housed in a controlled, arthropod-free University Animal Facility. B. henselae bacteremia was detected in 89% of the 47 naturally infected cattery cats. A total of 132 fleas were removed from cats whose blood was simultaneously cultured during different seasons and were tested individually for the presence of B. henselae DNA by PCR. B. henselae DNA was detected in 34% of 132 fleas, with seasonal variation, but without an association between the presence or the level of bacteremia in the corresponding cat. Cat fleas removed from bacteremic cattery cats transmitted B. henselae to five SPF kittens in two separate experiments; however, control SPF kittens housed with highly bacteremic kittens in the absence of fleas did not become infected. These data demonstrate that the cat flea readily transmits B. henselae to cats. Control of feline infestation with this arthropod vector may provide an important strategy for the prevention of infection of both humans and cats.
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              Canine and feline vector-borne diseases in Italy: current situation and perspectives

              In Italy, dogs and cats are at risk of becoming infected by different vector-borne pathogens, including protozoa, bacteria, and helminths. Ticks, fleas, phlebotomine sand flies, and mosquitoes are recognized vectors of pathogens affecting cats and dogs, some of which (e.g., Anaplasma phagocytophilum, Borrelia burgdorferi, Dipylidium caninum, Leishmania infantum, Dirofilaria immitis, and Dirofilaria repens) are of zoonotic concern. Recent studies have highlighted the potential of fleas as vectors of pathogens of zoonotic relevance (e.g., Rickettsia felis) in this country. While some arthropod vectors (e.g., ticks and fleas) are present in certain Italian regions throughout the year, others (e.g., phlebotomine sand flies) are most active during the summer season. Accordingly, control strategies, such as those relying on the systematic use of acaricides and insecticides, should be planned on the basis of the ecology of both vectors and pathogens in different geographical areas in order to improve their effectiveness in reducing the risk of infection by vector-borne pathogens. This article reviews the current situation and perspectives of canine and feline vector-borne diseases in Italy.
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                Author and article information

                Contributors
                linh.nguyen@uniba.it
                vito.colella@unimelb.edu.au
                grazia.greco@uniba.it
                fang8730@163.com
                wisnu-nc@ugm.ac.id
                upikke@gmail.com
                vmventurinadvm@clsu.edu.ph
                drkennethtong@gmail.com
                yltsai@mail.npust.edu.tw
                tpiyanan@hotmail.com
                saruda.t@cmu.ac.th
                sahatchai.t@cmu.ac.th
                lqthong@hcmuaf.edu.vn
                bklinh5@gmail.com
                thanhthomdo@gmail.com
                malaikawatanabe@gmail.com
                azaziah@upm.edu.my
                fdtvet@gmail.com
                lenaig.halos@boehringer-ingelheim.com
                frederic.beugnet@boehringer-ingelheim.com
                domenico.otranto@uniba.it
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                15 August 2020
                15 August 2020
                2020
                : 13
                : 420
                Affiliations
                [1 ]GRID grid.7644.1, ISNI 0000 0001 0120 3326, Dipartimento di Medicina Veterinaria, , Università degli Studi di Bari, ; Bari, Italy
                [2 ]GRID grid.1008.9, ISNI 0000 0001 2179 088X, Faculty of Veterinary and Agricultural Sciences, , University of Melbourne, ; Melbourne, Australia
                [3 ]GRID grid.256609.e, ISNI 0000 0001 2254 5798, School of Animal Science and Technology, , Guangxi University, ; Nanning, China
                [4 ]GRID grid.8570.a, Faculty of Veterinary Medicine, , Gadjah Mada University, ; Yogyakata, Indonesia
                [5 ]GRID grid.440754.6, ISNI 0000 0001 0698 0773, Faculty of Veterinary Medicine, , IPB University, ; Bogor, Indonesia
                [6 ]GRID grid.443260.7, ISNI 0000 0001 0664 3873, College of Veterinary Science & Medicine, , Central Luzon State University, ; Nueva Ecija, Philippines
                [7 ]Animal & Avian Veterinary Clinic, Singapore, Singapore
                [8 ]GRID grid.412083.c, ISNI 0000 0000 9767 1257, Department of Veterinary Medicine, , National Pingtung University of Science and Technology, ; Pingtung, Taiwan
                [9 ]Faculty of Veterinary Science, Chualalongkorn University, Bangkok, Thailand
                [10 ]GRID grid.7132.7, ISNI 0000 0000 9039 7662, Faculty of Veterinary Medicine, , Chiang Mai University, ; Chiang Mai, Thailand
                [11 ]GRID grid.444835.a, ISNI 0000 0004 0427 4789, Faculty of Animal Science & Veterinary Medicine, , Nong Lam University, ; Ho Chi Minh city, Vietnam
                [12 ]GRID grid.444964.f, ISNI 0000 0000 9825 317X, Faculty of Veterinary Medicine, , Vietnam National University of Agriculture, ; Hanoi, Vietnam
                [13 ]Biodiversity Conservation and Tropical Disease Research Institute, Hanoi, Vietnam
                [14 ]GRID grid.11142.37, ISNI 0000 0001 2231 800X, Faculty of Veterinary Medicine, , Universiti Putra Malaysia, ; Serdang, Malaysia
                [15 ]Department of Immunology, Aggeu Magalhães Institute, Recife, Brazil
                [16 ]GRID grid.484445.d, ISNI 0000 0004 0544 6220, Boehringer Ingelheim Animal Health, ; Lyon, France
                [17 ]GRID grid.411807.b, ISNI 0000 0000 9828 9578, Faculty of Veterinary Sciences, , Bu-Ali Sina University, ; Hamedan, Iran
                Author information
                http://orcid.org/0000-0002-7518-476X
                Article
                4288
                10.1186/s13071-020-04288-8
                7429691
                32799914
                885b0e49-f091-4543-8eff-4fa4e36c6d6d
                © The Author(s) 2020

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

                History
                : 12 June 2020
                : 3 August 2020
                Categories
                Research
                Custom metadata
                © The Author(s) 2020

                Parasitology
                ticks,fleas,dogs,cats,companion animals,asia,vector-borne pathogens,zoonotic
                Parasitology
                ticks, fleas, dogs, cats, companion animals, asia, vector-borne pathogens, zoonotic

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