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The immune response in infected dogs determines the clinical outcome, with a strong cell-mediated immune response linked to parasite control and mild clinical signs, while a humoral-dominant response is associated with severe disease. Low antibody levels in clinically asymptomatic dogs with negative molecular and\/or parasitological test results may reflect prior exposure or the early stages of <jats:italic>Leishmania<\/jats:italic> infection. In contrast, elevated antibody levels are typically correlated with a high parasitic burden and active disease. The detection of dogs with clinical leishmaniosis and null-specific immune response against <jats:italic>L. infantum<\/jats:italic> is uncommon. However, this presentation has also been described in human leishmaniasis with the absence of humoral response detected by conventional serological methods.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Case presentation<\/jats:title>\n            <jats:p>Case 1, a 9-year-old Border Collie, showed splenomegaly and <jats:italic>Leishmania<\/jats:italic> amastigotes within splenic macrophages. Case 2, a 10-month-old French Bulldog, had chronic anorexia and malabsorption syndrome with granulomatous splenitis and amastigotes confirmed by immunohistochemistry. Finally, case 3, a 7-year-old cross-breed, presented with cutaneous nodules and nasal ulcerative dermatitis, with <jats:italic>Leishmania<\/jats:italic> amastigotes detected histologically and confirmed by immunohistochemistry. All dogs were seronegative by two quantitative serological tests including indirect immunofluorescent test and enzyme-linked immunosorbent assay. The identification of the parasite in the affected organ established a clear cause-and-effect relationship. Consequently, anti-<jats:italic>Leishmania<\/jats:italic> treatment was initiated, consisting of allopurinol (10\u00a0mg\/kg orally twice daily) and meglumine antimoniate (50\u00a0mg\/kg subcutaneously twice daily for four weeks) in cases 1 and 3. In case 1, a favourable clinical response was noted, with a normal abdominal ultrasound and a negative result by quantitative molecular test from material obtained via ultrasound-guided splenic puncture. In case 3, the administration of meglumine antimoniate resulted in the resolution of dermatological signs. Clinical follow-up and anti-<jats:italic>Leishmania<\/jats:italic> treatment could not be performed for case 2.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>These findings highlight the diagnostic challenges in detecting clinical leishmaniosis in seronegative dogs. The absence of a specific humoral response should be considered, emphasizing the importance of using multiple diagnostic methods, including cytology, and histopathology with immunohistochemistry. This case series underscores the need for a comprehensive approach in diagnosing and managing canine leishmaniosis.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/s13028-025-00814-9","type":"journal-article","created":{"date-parts":[[2025,6,8]],"date-time":"2025-06-08T19:02:00Z","timestamp":1749409320000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Abscence of specific humoral response in three dogs with clinical leishmaniosis"],"prefix":"10.1186","volume":"67","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6209-4282","authenticated-orcid":false,"given":"Sergio","family":"Villanueva-Saz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diana","family":"Marteles","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1mparo","family":"Ortu\u00f1ez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda C.","family":"Ace\u00f1a","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janine E.","family":"Davies","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Riera","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda","family":"Borobia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maite","family":"Verde","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1lex","family":"G\u00f3mez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,6,8]]},"reference":[{"key":"814_CR1","doi-asserted-by":"publisher","first-page":"324","DOI":"10.1016\/j.pt.2008.04.001","volume":"24","author":"G Baneth","year":"2008","unstructured":"Baneth G, Koutinas AF, Solano-Gallego L, Bourdeau P, Ferrer L. 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