{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T09:35:58Z","timestamp":1769506558920,"version":"3.49.0"},"reference-count":38,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,21]],"date-time":"2025-09-21T00:00:00Z","timestamp":1758412800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Portuguese Foundation for Science and Technology (FCT)","award":["UI\/00772"],"award-info":[{"award-number":["UI\/00772"]}]},{"name":"Portuguese Foundation for Science and Technology (FCT)","award":["LA\/P\/0059\/2020"],"award-info":[{"award-number":["LA\/P\/0059\/2020"]}]},{"name":"Portuguese Foundation for Science and Technology (FCT)","award":["UIDB\/00511\/2020"],"award-info":[{"award-number":["UIDB\/00511\/2020"]}]},{"name":"Portuguese Foundation for Science and Technology (FCT)","award":["UIDP\/00511\/2020"],"award-info":[{"award-number":["UIDP\/00511\/2020"]}]},{"name":"Portuguese Foundation for Science and Technology (FCT)","award":["LA\/P\/0045\/2020"],"award-info":[{"award-number":["LA\/P\/0045\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Leishmania infantum is the causative agent of canine leishmaniosis (CanL), a zoonotic disease of considerable clinical and epidemiological concern. Quantification of parasite load is valuable for clinical management, particularly in low-parasite-load cases. This pilot study evaluated the correlation between cytological and molecular techniques in bone marrow samples from dogs clinically affected with leishmaniosis. Amastigotes were quantified by two independent observers using light microscopy, and the same samples were analysed by real-time polymerase chain reaction (qPCR) employing Leishmania spp. specific commercial primers. Inter-observer agreement was moderate according to Cohen\u2019s kappa (\u03ba = 0.47) and substantial according to the intraclass correlation coefficient (ICC = 0.63), respectively, confirming measurement reproducibility. A very strong inverse correlation was found between parasite counts and qPCR cycle threshold (Ct) values (Spearman\u2019s \u03c1 = \u22120.89; p &lt; 0.001). Furthermore, a robust logarithmic relationship (amastigotes\/\u03bcL = 10(\u22120.158 \u00d7 Ct + 7.61); R2 = 0.99994) was established allowing direct estimation of in vivo parasite concentration from molecular data. These preliminary findings suggest that qPCR can serve as a reliable, semi-quantitative tool, offering higher sensitivity in subclinical infections. The integration of molecular quantification with cytological methods enhances diagnostic accuracy and supports personalised therapeutic decision-making, advancing clinical management of CanL in a One Health context.<\/jats:p>","DOI":"10.3390\/microorganisms13092211","type":"journal-article","created":{"date-parts":[[2025,9,22]],"date-time":"2025-09-22T08:37:39Z","timestamp":1758530259000},"page":"2211","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["When Precision Matters: Bone Marrow Cytology Meets qPCR in a Pilot Study Quantifying Leishmania infantum Load in Dogs"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-2762-589X","authenticated-orcid":false,"given":"Juliana","family":"Costa","sequence":"first","affiliation":[{"name":"Department of Chemical and Biological Engineering (DEQB), Faculty of Engineering, University of Porto (FEUP), Rua Dr. Roberto Frias, 4200-465 Porto, Portugal"},{"name":"Molecular Diagnostics Laboratory, Unilabs Portugal, Centro Empresarial Lionesa Porto, Rua Lionesa, 4465-671 Le\u00e7a do Balio, Portugal"},{"name":"CEDIVET Veterinary Laboratories, Lionesa Business Hub, R. Lionesa 446 C24, 4465-671 Le\u00e7a do Balio, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-7587-3922","authenticated-orcid":false,"given":"Ana Rita","family":"Silva","sequence":"additional","affiliation":[{"name":"Molecular Diagnostics Laboratory, Unilabs Portugal, Centro Empresarial Lionesa Porto, Rua Lionesa, 4465-671 Le\u00e7a do Balio, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0206-6903","authenticated-orcid":false,"given":"Filipe","family":"Sampaio","sequence":"additional","affiliation":[{"name":"CEDIVET Veterinary Laboratories, Lionesa Business Hub, R. Lionesa 446 C24, 4465-671 Le\u00e7a do Balio, Portugal"},{"name":"Cytology and Haematology Diagnostic Services, Laboratory of Histology and Embryology, Department of Microscopy, ICBAS-School of Medicine and Biomedical Sciences, University of Porto (U.Porto), Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7022-5960","authenticated-orcid":false,"given":"Ana Patr\u00edcia","family":"Alves","sequence":"additional","affiliation":[{"name":"Molecular Diagnostics Laboratory, Unilabs Portugal, Centro Empresarial Lionesa Porto, Rua Lionesa, 4465-671 Le\u00e7a do Balio, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8098-7401","authenticated-orcid":false,"given":"Hugo","family":"Silva","sequence":"additional","affiliation":[{"name":"Molecular Diagnostics Laboratory, Unilabs Portugal, Centro Empresarial Lionesa Porto, Rua Lionesa, 4465-671 Le\u00e7a do Balio, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-5711-1905","authenticated-orcid":false,"given":"Hugo Lima","family":"de Carvalho","sequence":"additional","affiliation":[{"name":"CEDIVET Veterinary Laboratories, Lionesa Business Hub, R. Lionesa 446 C24, 4465-671 Le\u00e7a do Balio, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0708-6874","authenticated-orcid":false,"given":"Carlos","family":"Sousa","sequence":"additional","affiliation":[{"name":"Molecular Diagnostics Laboratory, Unilabs Portugal, Centro Empresarial Lionesa Porto, Rua Lionesa, 4465-671 Le\u00e7a do Balio, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3355-4398","authenticated-orcid":false,"given":"Manuel","family":"Sim\u00f5es","sequence":"additional","affiliation":[{"name":"LEPABE\u2014Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, Department of Chemical and Biological Engineering, University of Porto (U.Porto), 4200-465 Porto, Portugal"},{"name":"ALiCE\u2014Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto (U.Porto), 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5031-679X","authenticated-orcid":false,"given":"C\u00e1tia","family":"Fernandes","sequence":"additional","affiliation":[{"name":"Anicura Santa Marinha Veterinary Hospital, R. Dom Henrique de Cernache 183, 4400-625 Vila Nova de Gaia, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8182-5674","authenticated-orcid":false,"given":"Ana Patr\u00edcia","family":"Lopes","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Animal and Veterinary Research Centre (CECAV), Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7940-3141","authenticated-orcid":false,"given":"Andreia","family":"Garc\u00eas","sequence":"additional","affiliation":[{"name":"Animal and Veterinary Research Centre (CECAV), Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Wildlife Rehabilitation Centre (CRAS), Veterinary Teaching Hospital, University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), 5000-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0589-3533","authenticated-orcid":false,"given":"Elsa Leclerc","family":"Duarte","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, School of Science and Technology, University of \u00c9vora, Polo da Mitra, Apartado 94, 7002-554 \u00c9vora, Portugal"},{"name":"Mediterranean Institute for Agriculture, Environment and Development (MED), Global Change and Sustainability Institute (CHANGE), University of \u00c9vora, Polo da Mitra, Apartado 94, 7002-554 \u00c9vora, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7196-4179","authenticated-orcid":false,"given":"Ana Cl\u00e1udia","family":"Coelho","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Animal and Veterinary Research Centre (CECAV), Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6145-7560","authenticated-orcid":false,"given":"Lu\u00eds","family":"Cardoso","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Animal and Veterinary Research Centre (CECAV), Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9339-9932","authenticated-orcid":false,"given":"Ricardo","family":"Lopes","sequence":"additional","affiliation":[{"name":"Molecular Diagnostics Laboratory, Unilabs Portugal, Centro Empresarial Lionesa Porto, Rua Lionesa, 4465-671 Le\u00e7a do Balio, Portugal"},{"name":"CEDIVET Veterinary Laboratories, Lionesa Business Hub, R. Lionesa 446 C24, 4465-671 Le\u00e7a do Balio, Portugal"},{"name":"Department of Veterinary Sciences, University of Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Department of Veterinary and Animal Sciences, University Institute of Health Sciences (IUCS), CESPU, 4585-116 Gandra, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Priolo, V., Ippolito, D., Rivas-Estanga, K., De Waure, C., and Mart\u00ednez-Orellana, P. (2024). Canine leishmaniosis global prevalence over the last three decades: A meta-analysis and systematic review. Comp. Immunol. Microbiol. Infect. Dis., 112.","DOI":"10.1016\/j.cimid.2024.102211"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Lopes, R., Garc\u00eas, A., Silva, A., Brilhante-Sim\u00f5es, P., Martins, \u00c2., Duarte, E.L., Coelho, A.C., and Cardoso, L. (2024). Distribution of and relationships between epidemiological and clinicopathological parameters in canine leishmaniosis: A retrospective study of 15 years (2009\u20132023). Pathogens, 13.","DOI":"10.3390\/pathogens13080635"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1777","DOI":"10.1007\/s11259-023-10171-5","article-title":"Is leishmaniasis the new emerging zoonosis in the world?","volume":"47","author":"Llobat","year":"2023","journal-title":"Vet. Res. Commun."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1007\/s11250-024-03978-0","article-title":"Alternative hosts of Leishmania infantum: A neglected parasite in Europe","volume":"56","author":"Tsakmakidis","year":"2024","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Azami-Conesa, I., G\u00f3mez-Mu\u00f1oz, M.T., and Mart\u00ednez-D\u00edaz, R.A. (2021). A systematic review (1990-2021) of wild animals infected with zoonotic Leishmania. Microorganisms, 9.","DOI":"10.3390\/microorganisms9051101"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1645","DOI":"10.3390\/microbiolres15030109","article-title":"A systematic assessment of Leishmania donovani infection in domestic and wild animal reservoir hosts of zoonotic visceral leishmaniasis in India","volume":"15","author":"Ghouse","year":"2024","journal-title":"Microbiol. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1186\/s13071-024-06419-x","article-title":"Feline leishmaniosis in the mediterranean basin: A multicenter study","volume":"17","author":"Carbonara","year":"2024","journal-title":"Parasites Vectors"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Foglia Manzillo, V., Gizzarelli, M., Vitale, F., Montagnaro, S., Torina, A., Sotera, S., and Oliva, G. (2018). Serological and entomological survey of canine leishmaniasis in Lampedusa island, Italy. BMC Vet. Res., 14.","DOI":"10.1186\/s12917-018-1606-x"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1016\/S0020-7519(03)00142-5","article-title":"Two separate growth phases during the development of Leishmania in sand flies: Implications for understanding the life cycle","volume":"33","author":"Gossage","year":"2003","journal-title":"Int. J. Parasitol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Cec\u00edlio, P., Cordeiro-da-Silva, A., and Oliveira, F. (2022). Sand flies: Basic information on the vectors of leishmaniasis and their interactions with Leishmania parasites. Commun. Biol., 5.","DOI":"10.1038\/s42003-022-03240-z"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Cec\u00edlio, P., P\u00e9rez-Cabezas, B., Santar\u00e9m, N., Maciel, J., Rodrigues, V., and Cordeiro da Silva, A. (2014). Deception and manipulation: The arms of Leishmania, a successful parasite. Front. Immunol., 5.","DOI":"10.3389\/fimmu.2014.00480"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1186\/1756-3305-5-276","article-title":"Leishmania development in sand flies: Parasite-vector interactions overview","volume":"5","author":"Volf","year":"2012","journal-title":"Parasites Vectors"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Morales-Yuste, M., Mart\u00edn-S\u00e1nchez, J., and Corpas-Lopez, V. (2022). Canine leishmaniasis: Update on epidemiology, diagnosis, treatment, and prevention. Vet. Sci., 9.","DOI":"10.3390\/vetsci9080387"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1016\/j.tvjl.2014.09.002","article-title":"An update on the diagnosis and treatment of canine leishmaniosis caused by Leishmania infantum (Syn. L.\u2009chagasi)","volume":"202","author":"Noli","year":"2014","journal-title":"Vet. J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.actatropica.2014.08.016","article-title":"Comparison of PCR with stained slides of bone marrow and lymph nodes aspirates with suspect diagnosis for leishmaniasis","volume":"140","author":"Santos","year":"2014","journal-title":"Acta Trop."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Boggild, A.K., Valencia, B.M., Veland, N., Pilar Ramos, A., Calderon, F., Arevalo, J., Low, D.E., and Llanos-Cuentas, A. (2011). Non-invasive cytology brush PCR diagnostic testing in mucosal leishmaniasis: Superior performance to conventional biopsy with histopathology. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0026395"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3147","DOI":"10.1128\/JCM.00372-14","article-title":"Molecular tools for diagnosis of visceral leishmaniasis: Systematic review and meta-analysis of diagnostic test accuracy","volume":"52","author":"Leeflang","year":"2014","journal-title":"J. Clin. Microbiol."},{"key":"ref_18","first-page":"86","article-title":"Surgical approach of the anatomical sites for bone marrow aspiration in dogs","volume":"70","author":"Hesham","year":"2021","journal-title":"Alex. J. Vet. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Banko, A., Petrovic, G., Miljanovic, D., Loncar, A., Vukcevic, M., Despot, D., and Cirkovic, A. (2021). Comparison and sensitivity evaluation of three different commercial real-time quantitative PCR Kits for SARS-CoV-2 Detection. Viruses, 13.","DOI":"10.3390\/v13071321"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Nyaruaba, R., Mwaliko, C., Dobnik, D., Neu\u017eil, P., Amoth, P., Mwau, M., Yu, J., Yang, H., and Wei, H. (2022). Digital PCR applications in the SARS-CoV-2\/COVID-19 era: A roadmap for future outbreaks. Clin. Microbiol. Rev., 35.","DOI":"10.1128\/cmr.00168-21"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e81","DOI":"10.1093\/nar\/gkae683","article-title":"Advancing quantitative PCR with color cycle multiplex amplification","volume":"52","author":"Chen","year":"2024","journal-title":"Nucleic. Acids Res."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Jin, F., He, X., Zhang, J., Chen, L., Zhang, Y., Yang, Q., Liu, Z., Li, M., Ran, Y., and Guan, B.-O. (2025). A simple and integrated fiber-optic real-time qPCR platform for remote and distributed detection of epidemic virus infection. Biosens. Bioelectron., 273.","DOI":"10.1016\/j.bios.2025.117145"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Caballero-Solares, A., Hall, J.R., Xue, X., and Rise, M.L. (2022). Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) for gene expression analyses. Cancer Cell Biology: Methods and Protocols, Springer.","DOI":"10.1007\/978-1-0716-2376-3_21"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e0335422","DOI":"10.1128\/spectrum.03354-22","article-title":"Molecular diagnosis of leishmaniasis in Spain: Development and validation of ready-to-use gel-form nested and real-time PCRs to detect Leishmania spp.","volume":"11","author":"Chicharro","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Lopes, R., Sampaio, F., Carvalho, H.L.d., Garc\u00eas, A., Fernandes, C., Neves, C.V., Brito, A.S.d., Marques, T., Sousa, C., and Silva, A.R. (2024). Feline infectious peritonitis effusion index: A novel diagnostic method and validation of flow cytometry-based delta total nucleated cells analysis on the sysmex XN-1000V\u00ae. Vet. Sci., 11.","DOI":"10.3390\/vetsci11110563"},{"key":"ref_26","unstructured":"Petrie, A., and Watson, P. (2013). Statistics for Veterinary and Animal Science, Wiley-Blackwell. [3rd ed.]."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1007\/s10096-025-05037-4","article-title":"High detection rate of parasitic load by qPCR targeting 18s rDNA in blood of patients with active leishmaniasis lesions","volume":"44","author":"Castanheira","year":"2025","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1590\/S1984-29612013000300005","article-title":"Quantification of Leishmania infantum DNA in the bone marrow, lymph node and spleen of dogs","volume":"22","author":"Ramos","year":"2013","journal-title":"Rev. Bras. Parasitol. Vet."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1016\/j.actatropica.2009.05.008","article-title":"Molecular diagnosis of canine visceral leishmaniasis: Identification of Leishmania species by PCR-RFLP and quantification of parasite DNA by real-time PCR","volume":"111","author":"Quaresma","year":"2009","journal-title":"Acta Trop."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Oliveira, V.d.C., Junior, A.A.V.M., Ferreira, L.C., Calvet, T.M.Q., Dos Santos, S.A., Figueiredo, F.B., Campos, M.P., Rodrigues, F.d.C.d.C., de Oliveira, R.d.V.C., and de Lemos, E.R.S. (2021). Frequency of co-seropositivities for certain pathogens and their relationship with clinical and histopathological changes and parasite load in dogs infected with Leishmania infantum. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0247560"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Pantaleo, V., Furlanello, T., Campigli, M., Ventura, L., and Solano-Gallego, L. (2024). Short term treatment monitoring of renal and inflammatory biomarkers with naturally occurring leishmaniosis: A cohort study of 30 dogs. Vet. Sci., 11.","DOI":"10.3390\/vetsci11110517"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"621","DOI":"10.4269\/ajtmh.13-0376","article-title":"Bone marrow parasite burden among patients with New World Kala-Azar is associated with disease severity","volume":"90","author":"Silva","year":"2014","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1186\/s13071-017-2051-6","article-title":"Histological and parasitological distinctive findings in clinically-lesioned and normal-looking skin of dogs with different clinical stages of leishmaniosis","volume":"10","author":"Ordeix","year":"2017","journal-title":"Parasit Vectors"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2445","DOI":"10.1007\/s11010-023-04672-9","article-title":"Development of quantitative PCR and digital PCR for the quantification of Leishmania infantum in Dogs","volume":"478","author":"Pereira","year":"2023","journal-title":"Mol. Cell. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1186\/s13071-015-0770-0","article-title":"Usefulness of real time PCR to quantify parasite load in serum samples from chronic Chagas disease patients","volume":"8","author":"Melo","year":"2015","journal-title":"Parasit Vectors"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1186\/s13071-015-0909-z","article-title":"LeishVet update and recommendations on feline leishmaniosis","volume":"8","author":"Pennisi","year":"2015","journal-title":"Parasit Vectors"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1186\/1756-3305-4-86","article-title":"LeishVet guidelines for the practical management of canine leishmaniosis","volume":"4","author":"Koutinas","year":"2011","journal-title":"Parasit Vectors"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Verrest, L., Monnerat, S., Musa, A.M., Mbui, J., Khalil, E.A.G., Olobo, J., Wasunna, M., Chu, W.-Y., Huitema, A.D.R., and Schallig, H.D.F.H. (2024). Leishmania blood parasite dynamics during and after treatment of visceral leishmaniasis in eastern Africa: A pharmacokinetic-pharmacodynamic model. PLoS Negl. Trop. Dis., 18.","DOI":"10.1371\/journal.pntd.0012078"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/9\/2211\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:46:41Z","timestamp":1760035601000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/9\/2211"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,21]]},"references-count":38,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["microorganisms13092211"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms13092211","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,21]]}}}