Short Communication The genus Bartonella are fastidious, Gram-negative intracellular vector-borne bacteria with zoonotic potential. Cats have a crucial epidemiological role in the transmission of Bartonella henselae, the primary causal agent of cat-scratch disease in humans. Here we describe the first report of B. henselae infection in a cat in North Macedonia after a suspected Bartonella infection in a 7-year-old boy. A combination of cultural and molecular methods for detecting B. henselae in the asymptomatic cat was used. A blood sample was cultured on a blood agar plate, where colonies became visible after nine days of incubation. The DNA of a single colony was extracted, and the isolate was confirmed as B. henselae by sequencing the bacterial 16S ribosomal RNA gene and performing a Bartonella - specific PCR. The phylogenetic analysis demonstrated 100% similarity with B. henselae sequences, clustering on a single branch and establishing a distinct group. This finding indicates that B. henselae infection in cats circulates in North Macedonia and may affect feline and human health. https://macvetrev.mk/LoadArticlePdf/361 2023-9-27 207 213 10.2478/macvetrev-2023-0028 Bartonella henselae zoonosis public health detection Ivana Shikoska false 1 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia AUTHOR Igor Djadjovski false 2 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia AUTHOR Kiril Krstevski false 3 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia AUTHOR Zagorka Popova Hristovska false 4 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia AUTHOR Aleksandar Janevski false 5 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia AUTHOR Ljubica Rashikj false 6 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia AUTHOR Aleksandar Cvetkovikj acvetkovic@fvm.ukim.edu.mk false 7 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia LEAD_AUTHOR Iskra Cvetkovikj false 8 Veterinary Institute, Faculty of Veterinary Medicine-Skopje, Ss. Cyril and Methodius University in Skopje, Lazar Pop Trajkov 5-7, 1000 Skopje, North Macedonia AUTHOR Florin, T.A., Zaoutis, T.E., Zaoutis, L.B. (2008). Beyond cat scratch disease: Widening spectrum of Bartonella henselae infection. Pediatrics 121(5): e1413-e1425. PMid:18443019 1 https://doi.org/10.1542/peds.2007-1897 Chomel, B.B., Boulouis, H.J., Maruyama, S., Breitschwerdt, E.B. (2006). Bartonella spp. in pets and effect on human health. Emerg Infect Dis. 12(3): 389-394. PMid:16704774 PMCid:PMC3291446 2 https://doi.org/10.3201/eid1203.050931 Regier, Y., Ballhorn, W., Kempf, V.A.J. (2017). Molecular detection of Bartonella henselae in 11 Ixodes ricinus ticks extracted from a single cat. Parasit Vectors. 10(1): 105. PMid:28285589 PMCid:PMC5346845 3 https://doi.org/10.1186/s13071-017-2042-7 Wechtaisong, W., Bonnet, S.I., Lien, Y.Y., Chuang, S.T., Tsai, Y.L. (2020). Transmission of Bartonella henselae within Rhipicephalus sanguineus: Data on the potential vector role of the tick. PLoS Negl Trop Dis. 14(10): e0008664. PMid:33001978 PMCid:PMC7553266 4 https://doi.org/10.1371/journal.pntd.0008664 Álvarez-Fernández, A., Baxarias, M., Prandi, D., Breitschwerdt, E.B., Solano-Gallego, L. (2021). Bartonella henselae antibodies in serum and oral fluid specimens from cats. Pathogens 10(3): 329. PMid:33799577 PMCid:PMC8001045 5 https://doi.org/10.3390/pathogens10030329 Stepanić, M., Duvnjak, S., Reil, I., Špičić, S., Kompes, G., Beck, R. (2019). First isolation and genotyping of Bartonella henselae from a cat living with a patient with cat scratch disease in Southeast Europe. BMC Infect Dis. 19(1): 299. PMid:30940084 PMCid:PMC6446304 6 https://doi.org/10.1186/s12879-019-3929-z Poposki, K., Sopova, Z., Dimzova, M., Denkovska, E., Stoleska, T., Bosilkovski, M. (2020). Cat scratch disease: the first case report in Republic of North Macedonia. Pril (Makedon Akad Nauk Umet Odd Med Nauki). 41(3): 71-75. PMid:33500373 7 https://doi.org/10.2478/prilozi-2020-0048 Relman, D.A. (1993). Universal bacterial 16S rRNA amplification and sequencing. In D.H. Persing, T.F. Smith, F.C. Tenover, T.J. White (Eds.), Diagnostic molecular biology: principles and applications (pp. 489-495). Washington DC: ASM Press 8 Maggi, R.G., Diniz, P.P., Cadenas, M.B., Breitschwerdt, E.B. (2006). The use of molecular diagnostic techniques to detect Anaplasma, Bartonella and Ehrlichia species in arthropods or patients. The International Canine Vector-Borne Disease Symposium. August, 18-20, (pp. 9-14), Billesley, Alcester, UK 9 Schorn, S., Pfister, K., Reulen, H., Mahling, M., Silaghi, C. (2011). Occurrence of Babesia spp., Rickettsia spp. and Bartonella spp. in Ixodes ricinus in Bavarian public parks, Germany. Parasit Vectors. 4, 135. PMid:21762494 PMCid:PMC3154157 10 https://doi.org/10.1186/1756-3305-4-135 Staden Package [Internet]. A fully developed set of DNA sequence assembly (Gap4 and Gap5), editing and analysis tools (Spin) [cited 2023 June 30]. 11 https://staden.sourceforge.net/ National Library of Medicine [Internet]. Basic Local Alignment Search Tool (BLAST) [cited 2023 June 30]. 12 https://blast.ncbi.nlm.nih.gov/Blast.cgi Álvarez-Fernández, A., Breitschwerdt, E.B., Solano-Gallego, L. (2018). Bartonella infections in cats and dogs including zoonotic aspects. Parasit Vectors. 11, 624. PMid:30514361 PMCid:PMC6280416 13 https://doi.org/10.1186/s13071-018-3152-6 Kokkinaki, K.C.G., Saridomichelakis, M.N., Skampardonis, V., Mataragka, A., Ikonomopoulos, J., Leontides, L., Mylonakis, M.E., et al. (2022). Prevalence and risk factors for Bartonella spp. and haemoplasma infections in cats from Greece. Vet Sci. 9(7): 337. PMid:35878354 PMCid:PMC9315813 14 https://doi.org/10.3390/vetsci9070337 Guptill, L. (2010). Feline bartonellosis. Vet Clin North Am Small Anim Pract. 40(6): 1073-1090. PMid:20933137 15 https://doi.org/10.1016/j.cvsm.2010.07.009 Morelli, S., Crisi, P.E., Di Cesare, A., De Santis, F., Barlaam, A., Santoprete, G., Parrinello, C., et al. (2019). Exposure of client-owned cats to zoonotic vector-borne pathogens: clinic-pathological alterations and infection risk analysis. Comp Immunol Microbiol Infect Dis. 66, 101344. PMid:31437677 16 https://doi.org/10.1016/j.cimid.2019.101344 Breitschwerdt, E.B. (2008). Feline bartonellosis and cat scratch disease. Vet Immunol Immunopathol. 123(1-2): 167-171. PMid:18295347 17 https://doi.org/10.1016/j.vetimm.2008.01.025 Diddi, K., Chaudhry, R., Sharma, N., Dhawan, B. (2013). Strategy for identification & characterization of Bartonella henselae with conventional & molecular methods. Indian J Med Res. 137(2): 380-387. 18 Gutiérrez, R., Vayssier-Taussat, M., Buffet, J.P., Harrus, S. (2017). Guidelines for the isolation, molecular detection, and characterization of Bartonella species. Vector Borne Zoonotic Dis. 17(1): 42-50. PMid:28055575 19 https://doi.org/10.1089/vbz.2016.1956 Drummond, M.R., Lania, B.G., Diniz, P.P.V.P., Gilioli, R., Demolin, D.M.R., Scorpio, D.G., Breitschwerdt, E.B., Velho, P.E.N.F. (2018). Improvement of Bartonella henselae DNA detection in cat blood samples by combining molecular and culture methods. J Clin Microbiol. 56(5): e01732-17. PMid:29540455 PMCid:PMC5925714 20 https://doi.org/10.1128/JCM.01732-17 Duncan, A.W., Maggi, R.G., Breitschwerdt, E.B. (2007). A combined approach for the enhanced detection and isolation of Bartonella species in dog blood samples: Pre-enrichment liquid culture followed by PCR and subculture onto agar plates. J Microbiol Methods. 69(2): 273-281. PMid:17346836 21 https://doi.org/10.1016/j.mimet.2007.01.010 Saitou, N., Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 4(4): 406-425. 22 Tamura, K., Nei, M., Kumar, S. (2004). Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci U S A. 101(30): 11030-11035. PMid:15258291 PMCid:PMC491989 23 https://doi.org/10.1073/pnas.0404206101 Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39(4): 783-791. PMid:28561359 24 https://doi.org/10.1111/j.1558-5646.1985.tb00420.x Tamura, K., Stecher, G., Kumar, S. (2021). MEGA 11: molecular evolutionary genetics analysis version 11. Mol Biol Evol. 38(7): 3022-3027. PMid:33892491 PMCid:PMC8233496 25 https://doi.org/10.1093/molbev/msab120