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In the present work, we carried out a morphological, biometrical and molecular study of the species Archaeopsylla erinacei (Bouché, 1835) and their subspecies: Archaeopsylla erinacei erinacei (Bouché, 1835) and Archaeopsylla erinacei maura (Jordan & Rothschild, 1912) isolated from hedgehogs ( Erinaceus europaeus ) from different geographical regions (Seville and Corse). We have found morphological differences in females of A. erinacei from the same geographical origin that did not correspond with molecular differences. We suggest that some morphological characters traditionally used to discriminate females of both subspecies should be revised as well as we set the total length of the spermatheca as a valid criterion in order to discriminate between both subspecies. The Internal Transcribed Spacers 1 and 2 (ITS1, ITS2) and partial 18S rRNA gene, and partial cytochrome c-oxidase 1 ( cox1 ) and cytochrome b ( cytb ) mtDNA gene sequences were determined to clarify the taxonomic status of these taxa and to assess intra-specific and intra-population similarity. In addition, a phylogenetic analysis with other species of fleas using Bayesian and Maximum Likelihood analysis was performed. All molecular markers used, except 18S, showed molecular differences between populations corresponding with geographical origins. Thus, based on the phylogenetic and molecular study of two nuclear markers (ITS1, ITS2) and two mitochondrial markers ( cox1 and cytb ), as well as concatenated sequences of both subspecies, we reported the existence of two geographical genetic lineages in A. erinacei corresponding with two different subspecies: A. e. erinacei (Corse, France) and A. e. maura (Seville, Spain), that could be discriminated by polymerase chain reaction-linked random-fragment-length polymorphism.

Hedgehogs (e.g., Erinaceus europaeus, E. roumanicus) are wild mammals that frequently are observed near residential areas. The aim of this study was to investigate ticks and fleas collected from European hedgehogs in Portugal and to evaluate the prevalence of Rickettsia in those ectoparasites. Ticks and fleas were identified by morphological and molecular methods, and molecular detection by PCR and genotypic characterization of Rickettsia spp. was performed targeting ompB, ompA and gltA gene fragments. In total, 1892 ticks and 213 fleas were collected from 33 rescued European hedgehogs captured in seven districts of the north and centre of Portugal. Two tick species were identified – Rhipicephalus sanguineus accounted for 91 % (n = 1719) of the total ticks collected and 9 % (n = 173) were Ixodes hexagonus. All fleas were identified as Archaeopsylla erinacei. Regarding pathogen detection, Rickettsia massiliae DNA was found in 22 of the 212 tested Rh. sanguineus. None of the 48 I. hexagonus tested showed to be positive for rickettsiae. Rickettsia asembonensis DNA was identified in 55 A. erinacei fleas tested (n = 117). These results show that European hedgehogs are exposed to R. massiliae transmitted by ticks and to R. asembonensis via fleas suggesting that these mammals might be involved in the natural transmission cycle of these Rickettsia species. This study is the first report of R. asembonensis in fleas in Portugal.

Background In recent years, the scope and importance of emergent vector-borne diseases has increased dramatically. In Algeria, only limited information is currently available concerning the presence and prevalence of these zoonotic diseases. For this reason, we conducted a survey of hematophagous ectoparasites of domestic mammals and/or spleens of wild animals in El Tarf and Souk Ahras, Algeria. Methods Using real-time PCR, standard PCR and sequencing, the presence of Bartonella spp., Rickettsia spp., Borrelia spp. and Coxiella burnetii was evaluated in 268/1626 ticks, 136 fleas, 11 Nycteribiidae flies and 16 spleens of domestic and/or wild animals from the El Tarf and Souk Ahras areas. Results For the first time in Algeria, Bartonella tamiae was detected in 12/19 (63.2 %) Ixodes vespertilionis ticks, 8/11 (72.7 %) Nycteribiidae spp. flies and in 6/10 (60 %) bat spleens ( Chiroptera spp.). DNA from Coxiella burnetii , the agent of Q fever, was also identified in 3/19 (15.8 %) I. vespertilionis from bats. Rickettsia slovaca , the agent of tick-borne lymphadenopathy, was detected in 1/1 (100 %) Haemaphysalis punctata and 2/3 (66.7 %) Dermacentor marginatus ticks collected from two boars ( Sus scrofa algira ) respectively. Ri. massiliae , an agent of spotted fever, was detected in 38/94 (40.4 %) Rhipicephalus sanguineus sensu lato collected from cattle, sheep, dogs, boars and jackals. DNA of Ri. aeschlimannii was detected in 6/20 (30 %) Hyalomma anatolicum excavatum and 6/20 (30 %) Hy. scupense from cattle. Finally, Ri. felis , an emerging rickettsial pathogen, was detected in 80/110 (72.7 %) Archaeopsylla erinacei and 2/2 (100 %) Ctenocephalides felis of hedgehogs ( Atelerix algirus ). Conclusion In this study, we expanded knowledge about the repertoire of ticks and flea-borne bacteria present in ectoparasites and/or tissues of domestic and wild animals in Algeria.

Background The European hedgehog ( Erinaceus europaeus ) is known for high levels of ectoparasitism that not only represents a health risk for the animals themselves, but also for pet animals and humans as hedgehogs are frequently taken into human care. In the present study, patterns of ectoparasite infestation were assessed in hedgehogs taken into care at northern German animal rehabilitation centres. Methods Ectoparasites (ticks, fleas and mites) of 498 hedgehogs were collected over a period of 3 years from July 2018 to May 2021. Species were identified based on morphological characteristics and also via amplification and sequencing of the partial cytochrome c oxidase subunit 2 (COX-2) gene for fleas of the family Ceratophyllidae. Seasonal changes in infestation patterns as well as correlations with animal age, body weight and health status were assessed using generalised linear models. Results Infestation with ticks, fleas and mites occurred throughout the year. Overall, 86.5% (431/498) of the examined hedgehogs were infested with ticks, 91.4% (455/498) with fleas and 17.7% (88/498) with mites . Ixodes ricinus and Ixodes hexagonus / Ixodes canisuga were the most common tick species detected, with the additional occurrence of one Ixodes frontalis. Significant seasonal changes were observed for I. ricinus , but not for I. hexagonus / I. canisuga . Additionally, I. ricinus nymph prevalence declined significantly as of 2020, probably as a consequence of the climate change-related drought as of 2018. In hedgehogs with flea infestations, Archaeopsylla erinacei , Ceratophyllus sciurorum , Nosopsyllus fasciatus and Ctenocephalides felis were identified. In all cases of mite infestation, Caparinia tripilis was detected, in addition to specimens of the family Macronyssidae and free-living mites of the family Acaridae. Statistical analyses showed correlations regarding the factors month, year, body weight and age, but no correlation was evident regarding the health status of the animals. Conclusions With a detected infestation rate of 98.6%, almost all of the examined hedgehogs were infested with at least one ectoparasite species. The seasonal activity patterns of the different ectoparasite species together with the complex annual cycle of hedgehogs lead to different seasonal patterns in ectoparasite prevalence and infestation intensities. Due to the risk of transmission of zoonotic pathogens as well as the possible negative impact on the host itself, hedgehogs should be treated against ectoparasites when taken into care facilities. Graphical Abstract

Background Fleas are major ectoparasites of dogs and cats, with significant veterinary and public health implications. 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. Methods 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 c oxidase subunit II gene ( cox2 ) sequences. Epidemiological data were analysed through multivariate logistic regression models to identify possible risk factors associated with flea infestation. Results A total of 1052 dogs and 1039 cats were examined, and flea infestation prevalence was determined to be 33.6% and 36.5%, respectively. Ctenocephalides felis 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 C. felis felis . Other species identified included Ctenocephalides canis (9.6% in dogs; 1.8% in cats), Pulex irritans (4.2% in dogs) and Archaeopsylla erinacei maura (0.8% in dogs). The absence of insecticide use was the strongest predictor of flea infestation in both hosts (dogs: adjusted odds ratio [aOR] 4.87; cats: aOR 4.02). In dogs, the risk of infestation was higher in spring, summer and autumn compared to winter (aOR 2.08–3.72), and lower in the Lisbon Metropolitan Area, Alentejo and Algarve compared to the North region (aOR 0.14–0.45). In cats, risk was reduced in non-northern regions (Lisbon, Alentejo, Centro; aOR 0.10–0.45) and in those cats with non-domestic outdoor lifestyles (aOR 0.19). Conclusions Flea infestations are highly prevalent in dogs and cats across mainland Portugal, with C. felis felis as the dominant species. These findings provide valuable insights for the development of integrated, evidence-based strategies for flea control.

Hedgehogs are small omnivorous mammals prevalent across Europe, Asia, and Africa. The expanding intrusion of humans into hedgehog habitats and rising popularity of keeping exotic animals like hedgehogs as pets have disrupted the delicate balance of the human–animal–environment interface. Despite their ecological importance, hedgehog-borne flea species have received limited research attention, with most studies focusing on their vectorial role, and except one report, nothing on the species composition and genetic diversity of hedgehog flea specimens. An inventory and phylogenetic investigation, conducted for the first time in Algeria as well as in North Africa, allowed for collecting 45 hedgehogs (15 males and 30 females) and 303 fleas, and analyzing them through morphological and molecular approaches. The hedgehog specimens were subjected to macro- and microscopic examinations based on diagnostic morphological criteria and morphometric measurements, resulting in their identification as Atelerix algirus. This study represents the first report of this species’ prevalence in Djelfa. The ecological and geographical diversity of the studied areas, from arid deserts to humid regions, highlights the adaptability and resilience of A. algirus to diverse habitats. Additionally, 271 and 32 flea specimens were morphologically identified as Archaeopsylla erinacei and Ctenocephalides felis, respectively. Molecular examination targeting ITS1-rDNA further confirmed the morphological identification of fleas species. Phylogenetic analysis of fleas’ ITS1-rDNA sequences revealed heterogeneity, with the specimens clustering into two distinct clades. The first clade comprised two populations of A. erinacei, while the second clade included C. felis specimens of this study, grouped with homologous sequences from various regions worldwide. No correlation between flea species and geographical locations was observed demonstrating the sympatric distribution of flea specimens in the studied regions. In addition, no hybrid or genetic combination notion was observed among the flea specimens processed in this study compared with those coming from other countries. These findings contribute to our understanding of the species composition and distribution of hedgehogs and their flea ectoparasites in Algeria and provide a baseline for future epidemiological and entomological research in the country.

Fleas are ectoparasites of various animals, including Homo sapiens Linnaeus, 1758 (Primates: Hominidae). Among the species relevant to the human health field, either due to their dermatopathological potential or because of their role as vectors of microorganisms responsible for infectious diseases, such as plague or murine typhus, are the human flea, oriental rat flea, closely related cat and dog fleas, and chigoe flea. However, other species can accidentally infest humans. We have herein reported two unusual cases of humans infested and bitten by Archaeopsylla erinacei, the hedgehog flea. This species has been identified using stereomicroscopy, on the base of key characteristics. Furthermore, a brief literature review has revealed that hedgehog fleas could carry human-infectious agents, such as Rickettsia felis Bouyer et al. 2001 (Rickettsiales: Rickettsiaceae) or Bartonella henselae Regnery et al.1992 (Rhizobia les: Bartonellaceae). Using molecular biology, we thus tested nine A. erinacei specimens taken from these patients, for several bacteria species commonly associated with hematophagous arthropods, implicated in human pathology. However, all our samples were proven negative. The role of A. erinacei in human epidemiology has never been evaluated to date. This report sought to remind us that these fleas can be accidental parasites in humans. In addition, recent findings pertaining to bacteria of medical interest that are present in these insects should be brought to the fore, given that the question of their role as vectors in human infections remains unanswered and deserves further investigation.

Red foxes (Vulpes vulpes (L.)) are widespread across Europe, tolerant of synanthropic ecosystems, and susceptible to diseases potentially shared with humans and other animals. We describe flea fauna on red foxes in Romania, a large, ecologically diverse country, in part because fleas may serve as an indicator of the risk of spillover of vector-borne disease. We found 912 individual fleas of seven species on the 305 foxes assessed, for an infestation prevalence of 49.5%. Mean flea load per fox was 5.8 (range 0–44 fleas), and flea detections were most abundant in fall and early spring. Fleas included generalists (Ctenocephalides canis (Curtis), 32.6% of all fleas), Ct. felis (Bouché , 0.1%), and Pulex irritans L. (29.9%), the fox specialist Chaetopsylla globiceps (Taschenberg, 32.5%), mesocarnivore fleas Paraceras melis Walker (3.2%) and Ch. trichosa Kohaut (1.5%), and the small mammal flea Ctenophthalmus assimilis (Taschenberg, 0.1%), which is rarely or never reported from carnivores. There were significantly more female than male Ch. globiceps, Ct. canis, and Pu. irritans, and these three species were the most broadly distributed geographically. Diversity indices suggested reduced diversity in mountainous areas above 700 m. When compared to other flea studies on foxes in Europe, Romania had flea diversity near the median of reports, which was unexpected given Romania's high ecological diversity. Notably absent prey specialists, compared to other studies, include Archaeopsylla erinacei (Bouché) and Spilopsyllus cuniculi (Dale). Further studies of possible disease agents in fox fleas could help elucidate possible risks of vector-borne disease in foxes, domestic animals, and humans as well.

Small wild mammals are an important element in the emergence and transmission of vector-borne pathogens (VBPs). Among these species, hedgehogs have been found to be a reservoir of VBPs and host of arthropod vectors. Surveillance of VBPs in wildlife and their arthropods are crucial in a one health context. We conducted an exploratory study to screen Atelerix algirus hedgehogs and their infesting ticks and fleas for VBPs using a high throughput microfluidic real-time PCR system. Tested biopsies from hedgehogs were found to be naturally infected by Theileria youngi, Hepatozoon sp., Ehrlichia ewingii, Coxiella burnetii, and Candidatus Ehrlichia shimanensis. Similarly, Haemaphysalis erinacei and Rhipicephalus sanguineus tick species were infected by Ehrlichia ewingii, Rickettsia spp., Rickettsia massiliae, Borrelia sp., Coxiella burnetii, Rickettsia lusitaniae and Anaplasma sp. Archaeopsylla erinacei fleas were infected by Rickettsia asembonensis, Coxiella burnetii, and Rickettsia massiliae. Co-infections by two and three pathogens were detected in hedgehogs and infesting ticks and fleas. The microfluidic real-time PCR system enabled us not only to detect new and unexpected pathogens, but also to identify co-infections in hedgehogs, ticks, and fleas. We suggest that hedgehogs may play a reservoir role for VBPs in Tunisia and contribute to maintaining enzootic pathogen cycles via arthropod vectors.

Fleas are ectoparasitic insects of many terrestrial mammals and birds. To date, a limited number of parasitological studies in Turkey have focused on fleas; and therefore, distributions and host associations of many flea species have been still poorly known. In the present study, we collected a small number (n= 678, 409 females, 269 males) of fleas from some wild animals in Sivas province of Turkey. Fleas were identified as Archaeopsylla erinacei erinacei (Boucheé 1835), Ctenocephalides canis (Curtis 1826), Leptopsylla (Leptopsylla) taschenbergi taschenbergi (Wagner 1898), Nosopsyllus (Nosopsyllus) fasciatus (Bosc 1800), Chaetopsylla (Arctopsylla) hyaenae (Kolenati 1846), Chaetopsylla (Chaetopsylla) trichosa Kohaut 1903 and Ischnopsyllus (Ischnopsyllus) simplex Rothschild 1906. Of them, C. (A.) hyaenae and I. (I.) simplex were reported for the first time in Turkey. In the present study, we also reported some new flea-host associations for Turkey.