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This study aimed to estimate the prevalence of flea infestation in dogs and cats in mainland Portugal, identify associated risk factors and perform morphological and molecular characterisation of flea specimens.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods<\/jats:title>\n            <jats:p>A cross-sectional study was conducted from March 2022 to February 2023 in mainland Portugal. Dogs and cats were screened for flea infestations at veterinary clinics and shelters following World Association for the Advancement of Veterinary Parasitology guidelines. Fleas were morphologically identified to species level, and a subset was characterised molecularly via phylogenetic analysis of the cytochrome<jats:italic> c<\/jats:italic> oxidase subunit II gene (<jats:italic>cox2<\/jats:italic>) sequences. Epidemiological data were analysed through multivariate logistic regression models to identify possible risk factors associated with flea infestation.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>A total of 1052 dogs and 1039 cats were examined, and flea infestation prevalence was determined to be 33.6% and 36.5%, respectively. <jats:italic>Ctenocephalides felis<\/jats:italic> was the predominant flea species in both hosts, accounting for 85.7% of fleas in dogs and 98.8% in cats, with molecular data confirming the subspecies <jats:italic>C. felis felis<\/jats:italic>. Other species identified included <jats:italic>Ctenocephalides canis<\/jats:italic> (9.6% in dogs; 1.8% in cats), <jats:italic>Pulex irritans<\/jats:italic> (4.2% in dogs) and <jats:italic>Archaeopsylla erinacei maura<\/jats:italic> (0.8% in dogs). The absence of insecticide use was the strongest predictor of flea infestation in both hosts (dogs: adjusted odds ratio [aOR]\u20094.87; cats: aOR\u20094.02). In dogs, the risk of infestation was higher in spring, summer and autumn compared to winter (aOR\u20092.08\u20133.72), and lower in the Lisbon Metropolitan Area, Alentejo and Algarve compared to the North region (aOR\u20090.14\u20130.45). In cats, risk was reduced in non-northern regions (Lisbon, Alentejo, Centro; aOR\u20090.10\u20130.45) and in those cats with non-domestic outdoor lifestyles (aOR\u20090.19).<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>Flea infestations are highly prevalent in dogs and cats across mainland Portugal, with <jats:italic>C. felis felis<\/jats:italic> as the dominant species. These findings provide valuable insights for the development of integrated, evidence-based strategies for flea control.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/s13071-025-06904-x","type":"journal-article","created":{"date-parts":[[2025,7,6]],"date-time":"2025-07-06T04:34:25Z","timestamp":1751776465000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Epidemiological and molecular characterisation of flea infestations in dogs and cats in mainland Portugal"],"prefix":"10.1186","volume":"18","author":[{"given":"Andr\u00e9","family":"Pereira","sequence":"first","affiliation":[]},{"given":"Adrian","family":"Cruz","sequence":"additional","affiliation":[]},{"given":"Teresa","family":"Novo","sequence":"additional","affiliation":[]},{"name":"FleaPT group","sequence":"additional","affiliation":[]},{"given":"Ana","family":"Cardoso","sequence":"additional","affiliation":[]},{"given":"Ana","family":"Oliva","sequence":"additional","affiliation":[]},{"given":"Ana","family":"R\u00f3is","sequence":"additional","affiliation":[]},{"given":"Ana","family":"Sarago\u00e7a","sequence":"additional","affiliation":[]},{"given":"Andr\u00e9","family":"Silva","sequence":"additional","affiliation":[]},{"given":"Andreia","family":"Pinto","sequence":"additional","affiliation":[]},{"given":"Aniana","family":"Correia","sequence":"additional","affiliation":[]},{"given":"Bruno","family":"Almeida","sequence":"additional","affiliation":[]},{"given":"Gon\u00e7alo","family":"Lixa","sequence":"additional","affiliation":[]},{"given":"Guida","family":"Brito","sequence":"additional","affiliation":[]},{"given":"Hugo","family":"Vilhena","sequence":"additional","affiliation":[]},{"given":"Jos\u00e9","family":"Chaves","sequence":"additional","affiliation":[]},{"given":"Karine","family":"Torres","sequence":"additional","affiliation":[]},{"given":"Lu\u00eds","family":"Martins","sequence":"additional","affiliation":[]},{"given":"Madalena","family":"Lemos","sequence":"additional","affiliation":[]},{"given":"Margarida","family":"C\u00e2mara","sequence":"additional","affiliation":[]},{"given":"Miguel","family":"Almas","sequence":"additional","affiliation":[]},{"given":"Patr\u00edcia","family":"Cachola","sequence":"additional","affiliation":[]},{"given":"Paulo","family":"Afonso","sequence":"additional","affiliation":[]},{"given":"Ricardo","family":"Dias","sequence":"additional","affiliation":[]},{"given":"Rita","family":"Calouro","sequence":"additional","affiliation":[]},{"given":"Ruth","family":"Gomes","sequence":"additional","affiliation":[]},{"given":"Sabrina","family":"Rodrigues","sequence":"additional","affiliation":[]},{"given":"Sara","family":"Lang","sequence":"additional","affiliation":[]},{"given":"Sofia","family":"Piteira","sequence":"additional","affiliation":[]},{"given":"Telma","family":"Gomes","sequence":"additional","affiliation":[]},{"given":"Carla","family":"Maia","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2025,7,6]]},"reference":[{"key":"6904_CR1","doi-asserted-by":"publisher","first-page":"e667","DOI":"10.1016\/j.ijid.2009.11.011","volume":"14","author":"I Bitam","year":"2010","unstructured":"Bitam I, Dittmar K, Parola P, Whiting MF, Raoult D. 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Animal participation was voluntary, requiring written informed consent from the legal detainer after reading an informational document about the study. Flea collection and animal handling were carried out by veterinarians, nurses, or technical assistants, ensuring minimal risk and no harm or distress to the animals.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"263"}}