BACKGROUND: Hippoboscid flies are bloodsucking arthropods that can transmit pathogenic microorganisms and are therefore potential vectors for pathogens such as Bartonella spp. These Gram-negative bacteria can cause mild-to-severe clinical signs in humans and animals; therefore, monitoring Bartonella spp. prevalence in louse fly populations appears to be a useful prerequisite for zoonotic risk assessment. METHODS: Using convenience sampling, we collected 103 adult louse flies from four ked species (Lipoptena cervi, n = 22; Lipoptena fortisetosa, n = 61; Melophagus ovinus, n = 12; Hippobosca equina, n = 8) and the pupae of M. ovinus (n = 10) in the federal state of Saxony, Germany. All the samples were screened by polymerase chain reaction (PCR) for Bartonella spp. DNA, targeting the citrate synthase gene (gltA). Subsequently, PCRs targeting five more genes (16S, ftsZ, nuoG, ribC and rpoB) were performed for representatives of revealed gltA genotypes, and all the PCR products were sequenced to identify the Bartonella (sub)species accurately. RESULTS AND CONCLUSIONS: The overall detection rates for Bartonella spp. were 100.0%, 59.1%, 24.6% and 75.0% in M. ovinus, L. cervi, L. fortisetosa and H. equina, respectively. All the identified bartonellae belong to the Bartonella schoenbuchensis complex. Our data support the proposed reclassification of the (sub)species status of this group, and thus we conclude that several genotypes of B. schoenbuchensis were detected, including Bartonella schoenbuchensis subsp. melophagi and Bartonella schoenbuchensis subsp. schoenbuchensis, both of which have previously validated zoonotic potential. The extensive PCR analysis revealed the necessity of multiple PCR approach for proper identification of the ruminant-associated bartonellae.

Department of Parasitology Faculty of Science Charles University Prague Czech Republic

Faculty of Agriculture Environment Chemistry HTW Dresden University of Applied Sciences Dresden Germany

Laboratory for Zoonoses and Environmental Microbiology National Institute for Public Health and the Environment Bilthoven The Netherlands

ZAFT e 5 Centre for Applied Research and Technology Dresden Germany

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Antequera‐Gómez, M. L. , Lozano‐Almendral, L. , Barandika, J. F. , González‐Martín‐Niño, R. M. , Rodríguez‐Moreno, I. , García‐Pérez, A. L. , & Gil, H. (2015). Bartonella chomelii is the most frequent species infecting cattle grazing in communal mountain pastures in Spain. Applied and Environmental Microbiology, 81(2), 623–629. PubMed PMC

Baltes‐Götz, B. (2012). Logistische Regressionsanalyse mit SPSS . SPSS.

Backhaus, K. , Erichson, B. , Plinke, W. , & Weiber, R. (2016). Multivariate analysemethoden. Springer.

Bartosik, K. , Maślanko, W. , Buczek, A. , Asman, M. , Witecka, J. , & Szwaj, E. (2021). Two new haplotypes of Bartonella sp. Isolated from Lipoptena fortisetosa (Diptera: Hippoboscidae) in SE Poland. Insects, 12(6), 485. PubMed PMC

Bezerra‐Santos, M. A. , & Otranto, D. (2020). Keds, the enigmatic flies and their role as vectors of pathogens. Acta Tropica, 209, 105521. PubMed

Boucheikhchoukh, M. , Mechouk, N. , Benakhla, A. , Raoult, D. , & Parola, P. (2019). Molecular evidence of bacteria in Melophagus ovinus sheep keds and Hippobosca equina forest flies collected from sheep and horses in Northeastern Algeria. Comparative Immunology, Microbiology and Infectious Diseases, 65, 103–109. PubMed

de Bruin, A. , van Leeuwen, A. D. , Jahfari, S. , Takken, W. , Földvári, M. , Dremmel, L. , Sprong, H. , & Földvári, G. (2015). Vertical transmission of Bartonella schoenbuchensis in Lipoptena cervi . Parasites & Vectors, 8(1), 1–6. PubMed PMC

Büttiker, W. (1994). Die Lausfliegen der Schweiz (Diptera, Hippoboscidae): Mit Bestimmungsschlüssel (Documenta faunistica Helvetiae). Centre suisse de cartographie de la faune. https://books.google.de/books?id=BsuePQAACAAJ

Chomel, B. B. , Kasten, R. W. , Sykes, J. E. , Boulouis, H.‐J. , & Breitschwerdt, E. B. (2003). Clinical impact of persistent Bartonella bacteremia in humans and animals. Annals of the New York Academy of Sciences, 990(1), 267–278. PubMed

Chung, C. Y. , Kasten, R. W. , Paff, S. M. , van Horn, B. A. , Vayssier‐Taussat, M. , Boulouis, H.‐J. , & Chomel, B. B. (2004). Bartonella spp. DNA associated with biting flies from California. Emerging Infectious Diseases, 10(7), 1311. PubMed PMC

Dehio, C. , Sauder, U. , & Hiestand, R. (2004). Isolation of Bartonella schoenbuchensis from Lipoptena cervi, a blood‐sucking arthropod causing deer ked dermatitis. Journal of Clinical Microbiology, 42(11), 5320–5323. PubMed PMC

do Amaral, R. B. , Cardozo, M. V. , Varani, A. M. , Furquim, M. E. C. , Dias, C. M. , Assis, W. O. , da Silva, A. R. , Herrera, H. M. , Machado, R. Z. , & André, M. R. (2022). First report of Bartonella spp. in Marsupials from Brazil, with a description of Bartonella harrusi sp. nov. and a new proposal for the taxonomic reclassification of species of the genus Bartonella . Microorganisms, 10(8), 1609. PubMed PMC

du Prel, J.‐B. , Hommel, G. , Röhrig, B. , & Blettner, M. (2009). Confidence interval or p‐value? Part 4 of a series on evaluation of scientific publications. Deutsches Ärzteblatt International, 106(19), 335–339. 10.3238/arztebl.2009.0335 PubMed DOI PMC

du Prel, J.‐B. , Röhrig, B. , Hommel, G. , & Blettner, M. (2010). Choosing statistical tests: Part 12 of a series on evaluation of scientific publications. Deutsches Ärzteblatt International, 107(19), 343. PubMed PMC

Duodu, S. , Madslien, K. , Hjelm, E. , Molin, Y. , Paziewska‐Harris, A. , Harris, P. D. , Colquhoun, D. J. , & Ytrehus, B. (2013). Bartonella infections in deer keds (Lipoptena cervi) and moose (Alces alces) in Norway. Applied and Environmental Microbiology, 79(1), 322–327. PubMed PMC

Eckert, J. (2008). Lehrbuch der parasitologie für die tiermedizin: 112 tabellen. Georg Thieme Verlag.

Halos, L. , Jamal, T. , Maillard, R. , Girard, B. , Guillot, J. , Chomel, B. , Vayssier‐Taussat, M. , & Boulouis, H.‐J. (2004). Role of Hippoboscidae flies as potential vectors of Bartonella spp. infecting wild and domestic ruminants. Applied and Environmental Microbiology, 70(10), 6302–6305. PubMed PMC

Hosmer, D. W. , Lemeshow, S. , & Sturdivant, R. X. (2000). Applied logistic regression. Wiley New York.

Iannino, F. , Salucci, S. , Di Provvido, A. , Paolini, A. , & Ruggieri, E. (2018). Bartonella infections in humans dogs and cats. Veterinaria Italiana, 54(1), 63–72. PubMed

Knauthe, C. , & Jentzsch, M. (2019). Die Lausfliegenfauna des Freistaats Sachsen (Díptera, Hippoboscidae). Entomologische Nachrichten und Berichte, 63, 189–196.

Korhonen, E. M. , Vera, C. P , Pulliainen, A. T. , Sironen, T. , Aaltonen, K. , Kortet, R. , Kortet, R. , Härkönen, L. , Härkönen, S. , Paakkonen, T. , Nieminen, P. , Mustonen, A.‐M. , Ylönen, H. , & Vapalahti, O. (2015). Molecular detection of Bartonella spp. in deer ked pupae, adult keds and moose blood in Finland. Epidemiology & Infection, 143(3), 578–585. PubMed PMC

Kosoy, M. , Bai, Y. , Enscore, R. , Rizzo, M. R. , Bender, S. , Popov, V. , Albayrak, L. , Fofanov, Y. , & Chomel, B. (2016). Bartonella melophagi in blood of domestic sheep (Ovis aries) and sheep keds (Melophagus ovinus) from the southwestern US: Cultures, genetic characterization, and ecological connections. Veterinary Microbiology, 190, 43–49. PubMed

Kumsa, B. , Parola, P. , Raoult, D. , & Socolovschi, C. (2014). Bartonella melophagi in Melophagus ovinus (sheep ked) collected from sheep in northern Oromia, Ethiopia. Comparative Immunology, Microbiology and Infectious Diseases, 37(1), 69–76. PubMed

Kurina, O. , Kirik, H. , Õunap, H. , & Õunap, E. (2019). The northernmost record of a blood‐sucking ectoparasite, Lipoptena fortisetosa Maa (Diptera: Hippoboscidae), in Estonia. Biodiversity Data Journal, 7, e47857. PubMed PMC

Lee, S.‐H. , Kim, K.‐T. , Kwon, O.‐D. , Ock, Y. , Kim, T. , Choi, D. , & Kwak, D. (2016). Novel detection of Coxiella spp., Theileria luwenshuni, and T. ovis endosymbionts in deer keds (Lipoptena fortisetosa). PloS One, 11(5), e0156727. PubMed PMC

Liu, Y. , He, B. , Li, F. , Li, K. , Zhang, L. , Li, X. , & Zhao, L. (2018). Molecular identification of Bartonella melophagi and Wolbachia supergroup F from sheep keds in Xinjiang, China. The Korean Journal of Parasitology, 56(4), 365. PubMed PMC

Liu, Y.‐H. , He, B. , Li, K.‐R. , Li, F. , Zhang, Lu‐Y. , Li, X.‐Q. , & Zhao, L. (2019). First report of border disease virus in Melophagus ovinus (sheep ked) collected in Xinjiang, China. PloS One, 14(8), e0221435. PubMed PMC

List of Prokaryotic Names with Standing in Nomenclature (LPSN) . (2023). Genus Bartonella. Accessed December 4, 2023. Available online: https://lpsn.dsmz.de/genus/bartonella

Müller, J. (2017). Aktualisierte und kommentierte Checkliste der Lausfliegen (Diptera: Hippoboscidae) Deutschlands. Studia Dipterologica, 24, 1–7.

National Center for Biotechnology Information (NCBI) . (2023). Genbank Database. Accessed December 4, 2023. Available online: https://www.ncbi.nlm.nih.gov/

Norman, A. F. , Regnery, R. , Jameson, P. , Greene, C. , & Krause, D. C. (1995). Differentiation of Bartonella‐like isolates at the species level by PCR‐restriction fragment length polymorphism in the citrate synthase gene. Journal of Clinical Microbiology, 33(7), 1797–1803. PubMed PMC

Okaro, U. , Addisu, A. , Casanas, B. , & Anderson, B. (2017). Bartonella species, an emerging cause of blood‐culture‐negative endocarditis. Clinical Microbiology Reviews, 30(3), 709–746. PubMed PMC

Reeves, W. K. , & Lloyd, J. E. (2019). Louse flies, keds, and bat flies (Hippoboscoidea). In Medical and veterinary entomology (pp. 421–438). Elsevier.

Regier, Y. , Komma, K. , Weigel, M. , Pulliainen, A. T. , Göttig, S. , Hain, T. , & Kempf, V. A. J. (2018). Microbiome analysis reveals the presence of Bartonella spp. and Acinetobacter spp. in deer keds (Lipoptena cervi). Frontiers in Microbiology, 9, 3100. PubMed PMC

Rudolf, I. , Betášová, L. , Bischof, V. , Kristýna, V. , Blažejová, H. , Mendel, J. , Hubálek, Z. , & Kosoy, M. (2016). Molecular survey of arthropod‐borne pathogens in sheep keds (Melophagus ovinus), Central Europe. Parasitology Research, 115(10), 3679–3682. PubMed

Sato, S. , Kabeya, H. , Ishiguro, S. , Shibasaki, Y. , & Maruyama, S. (2021). Lipoptena fortisetosa as a vector of Bartonella bacteria in Japanese sika deer (Cervus nippon). Parasites & Vectors, 14(1), 1–10. PubMed PMC

Small, R. W. (2005). A review of Melophagus ovinus (L.), the sheep ked. Veterinary Parasitology, 130(1–2), 141–155. PubMed

Szewczyk, T. , Werszko, J. , Steiner‐Bogdaszewska, Ż. , Jeżewski, W. , Laskowski, Z. , & Karbowiak, G. (2017). Molecular detection of Bartonella spp. in deer ked (Lipoptena cervi) in Poland. Parasites & Vectors, 10(1), 1–7. PubMed PMC

Victor, A. , Elsäßer, A. , Hommel, G. , & Blettner, M. (2011). Wie bewertet man die p‐Wert‐Flut? Hinweise zum Umgang mit dem multiplen testen—Teil 10 der Serie zur Bewertung wissenschaftlicher Publikationen. Deutsche Zahnarztliche Zeitschrift, 66(5), 366.

Diversity and Multiple Infections of Bartonella in Red Deer and Deer Keds