A case of mistaken identity: a systematic review, meta-analysis, and reinvestigation of hemotropic Mycoplasma spp. infection in Ctenocephalides felis (cat flea).

BACKGROUND: Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence. METHODS: A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed. RESULTS: Flea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining "hemoplasma infected" fleas, PCR amplified Spiroplasma or other bacteria. CONCLUSIONS: Therefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary.

Prophylaxis Against Heartworm Infection and Flea or Tick Infestation in Dogs: Single Agents and Combination-Drug Products.

From ancient times to the present, parasites and the diseases they transmit have jeopardized the health and wellbeing of working and companion canines worldwide. Many common pests that afflict dogs can be classified as ectoparasites (e.g., fleas, ticks, lice), which serve as vectors of pathogens transmitted as the organism feeds or defecates; or endoparasites (e. g, helminths, protozoa), which can cause slowly progressive subclinical canine diseases as well as acute illnesses associated with high morbidity and mortality rates. Safe, effective antiparasitic prophylaxis in dogs remains a topic of major interest to both veterinarians and their clients, especially with respect to the prevention of canine heartworm infection and flea or tick infestation. Many compounders, especially those whose pharmacy includes a retail component, counsel veterinarians and pet owners about preparations and commercially available medications that prevent or treat parasitic infestations and provide assistance in obtaining those therapies. To support such efforts, this article provides information about single agents and combination-drug products prophylactic against common canine parasites, emerging resistance to those medications, and the toxic effects that such treatments can engender in some canine breeds.

Functional similarity affects similarity in partner composition in flea-mammal networks.

Functional signal in an interaction network is a phenomenon in which species resembling each other in their traits interact with similar partners. We tested the functional signal concept in realm-specific and regional flea-host networks from four biogeographic realms and asked whether the species composition of (a) host spectra and (b) flea assemblages is similar between functionally similar flea and host species, respectively. Analogously to testing for phylogenetic signal, we applied Mantel tests to investigate the correlation between flea or host functional distances calculated from functional dendrograms and dissimilarities in sets of interacting partners. In all realm-specific networks, functionally similar fleas tended to exploit similar hosts often belonging to the same genus, whereas functionally similar hosts tended to harbour similar fleas, again often belonging to the same genus. The strength of realm-specific functional signals and the frequency of detecting a significant functional signal in the regional networks differed between realms. The frequency of detecting a significant functional signal in the regional networks correlated positively with the network size for fleas and with the number of hosts in a network for hosts. A functional signal in the regional networks was more frequently found for hosts than for fleas. We discuss the mechanisms behind the functional signal in both fleas and their hosts, relate geographic functional signal patterns to the historic biogeography of fleas and conclude that functional signals in the species composition of host spectra for fleas and of flea assemblages for hosts result from the interplay of evolutionary and ecological processes.

First record of a structural infestation by the bird flea Dasypsyllus gallinulae perpinnatus (Siphonaptera: Ceratophyllidae) in the San Francisco Bay Area, California, USA.

Herein we present the first-known case report of a structural infestation by the bird flea (Dasypsyllus gallinulae perpinnatus) (Dale 1878) in the United States. In March of 2023, the San Mateo County Mosquito and Vector Control District was contacted by a resident in South San Francisco, CA who reported the presence of fleas inside their condominium. The resident had 2 dogs who were on oral flea medication and only 1 inhabitant out of 4 reported receiving flea bites. The front walkway, backyard, and garage were flagged and a small passerine nest was removed from a fire alarm bell in the front walkway. A total of 31 fleas (13 males, 18 females) were collected by flagging from the front entryway underneath the nest. One female flea was collected from the backyard, and 20 (9 males, 11 females) were collected by the resident from inside the house. A total of 387 fleas (163 males, 224 females) were collected from the nest of a small passerine. All life stages (egg, larvae, pupae, and adult) were observed within the nest. Additional parasites in the nest included bird lice and larval western black-legged ticks (Ixodes pacificus, Cooley and Kohls 1943). Research should be conducted into whether bird fleas are capable of transmitting avian pathogens to humans and this information should be communicated to healthcare providers as part of a One Health approach. Additionally, resources to aid in species-level flea identification should be made accessible to pest control operators as this will aid the development of targeted treatments as part of an integrated pest management plan.

Molecular survey of flea-borne pathogens in fleas associated with carnivores from Algeria and an Artificial Neural Network-based risk analysis of flea-borne diseases.

As ectoparasites and efficient vectors of pathogens fleas constitute a source of nuisance for animals as well as a major issue for public health in Algeria. In this study, a molecular survey has been conducted to investigate the presence of pathogens in fleas infesting domestic and wild carnivores in the central north and eastern north and south of Algeria. The molecular screening that targeted Acanthocheilonema reconditum, Bartonella spp.,and Dipylidium caninum, was supplemented by a comprehensive analysis of risk factors related to flea-borne pathogens, drawing data from all documentation across multiple languages and sources from Morocco, Algeria, and Tunisia. In the current study, several Bartonella spp. 56/430 (13.02%) and Dipylidium caninum 3/430 (0.7%) were identified. The sequencing results revealed 5/23 (21.74%) B. clarridgeiae, 3/23 (13.04%) B. henselae, and 3/23 (13.04%) B. vinsonii. The two haplotypes, H1 and H2, of D. caninum were identified for the first time in North Africa. The results of the Artificial Neural Network risk analyses unveiled that the prevalence of pathogens and the presence of host generalist fleas as well as the vectorial competence are the most determinant risk factors of flea-borne diseases in Maghreb.

Fleas of wild carnivores in Romania: diversity, distribution, and host-associations.

BACKGROUND: Fleas are important hematophagous insects, infesting mammals and birds with a worldwide distribution. Fleas of medical importance have been reported from various carnivores worldwide, such as felids, canids, or mustelids. Romania hosts a wide carnivore diversity, but very little is known about flea species that parasitize these animals in Romania. This study aimed to provide a better understanding of the fleas' diversity and their distribution in a relatively large and diverse number of wild carnivore hosts from Romania. METHODS: From 2013 to 2021, 282 carcasses of wild carnivores from different locations in Romania were collected and examined for the presence of ectoparasites. All collected fleas were morphologically identified using specific keys and descriptions. An analysis of the co-occurrence networks was performed. RESULTS: A total of 11 flea species were identified: Pulex irritans (41.09%), Paraceras melis (20.11%), Ctenocephalides felis (7.33%), Ctenocephalides canis (7.83%), Monopsyllus sciurorum (11.11%), Chaetopsylla trichosa (21.96%), Chaetopsylla homoea (5.5%), Chaetopsylla tuberculaticeps (100%), Chaetopsylla rothschildi (13.33%), Chaetopsylla sp. (14.34%), Chaetopsylla globiceps (5.12%), Echidnophaga gallinacea (10%). The statistical analyses showed a significant difference between the infestation of Martes foina with females being more frequently infected than males (66% versus 33%). Paraceras melis infesting Meles meles had a significantly higher prevalence in female badgers than in males (× 2 = 7.7977, P < 0.01) and higher intensities of infestations in males than in females (t = 1.871, P < 0.05). CONCLUSIONS: This is the first large-scale study investigating the distribution and diversity of flea species infesting wild carnivores in Romania. Three flea species were identified for the first time in Romania (E. gallinacea, C. homoea, and C. tuberculaticeps).

Survey of ectoparasites affecting dog and cat populations living in sympatry in Gamo Zone, Southern Ethiopia.

BACKGROUND: Ectoparasites of dogs and cats are implicated to be responsible for life-threatening anaemia, allergic dermatitis and pruritic and non-pruritic skin disorders. In Ethiopia, few studies have been conducted on the arthropods of dogs and cats. OBJECTIVES: In order to shed light on some of these aspects, a survey was conducted to investigate the presence of ectoparasites in dogs and cats living in sympatry in the urban and rural areas of Gamo Zone, Ethiopia. METHODS: A total of 297 dogs and 110 cats were examined for ectoparasites, and questionnaires were employed to obtain information concerning owner's knowledge about arthropods and vector-borne diseases (VBDs). RESULTS: The overall prevalence of ticks, fleas and lice in dogs was 36.7%, 69.7% and 4.7%, respectively. Similarly, on cats, an overall prevalence of 2.7% ticks and 21.8% fleas was recorded. On dogs, fleas (Ctenocephalides felis 69.4%, Echidnophaga gallinacea 1.3%, Ctenocephalides canis 1.0% and Pulex irritans 0.3%), ticks (Amblyomma variegatum 22.9%, Rhipicephalus sanguineus 14.1%, Haemaphysalis leachi 8.8%, Rhipicephalus praetextatus 4.0% and Rhipicephalus pulchellus 3.4%) and lice (Heterodoxus spiniger 4.0% and Trichodectes canis 0.7%) were identified. Likewise, on cats, fleas (C. felis [15.5%] and E. gallinacea [7.3%]) and ticks (H. leachi [2.7%]) were identified. The abundance of C. felis was significantly higher (p < 0.001) on dogs, whereas in cats, the abundance of E. gallinacea was significantly higher (p = 0.002) than the other ectoparasites. On dogs, a significantly higher prevalence of Rh. sanguineus was recorded in urban areas (<0.001) and on dogs which live in indoor environments (p = 0.003) than on dogs which live in other environments. On the other hand, the prevalence of A. variegatum in rural areas and midland agroecology was significantly higher (p < 0.001). The prevalence of H. leachi was significantly higher in midland (p < 0.001) and on adult dogs (p = 0.001). Overall, fleas were more prevalent in rural (p = 0.029) than in urban areas, and female dogs were with higher infestation than the male (p = 0.047) dogs; C. felis was prevalent in female (p = 0.038) dogs than males. Overall, 88.3% owners in the study area had no knowledge about ectoparasites and VBDs of dogs and cats. Majority of the owners (64.8%) attest that they had never visited veterinary clinics. CONCLUSIONS: In conclusion, the data presented in the present study provide additional knowledge on the importance of ectoparasites of dogs and cats and are believed to contribute in awareness creation and strengthening of veterinary services of dogs and cats of the study area.

Relationships between functional alpha and beta diversities of flea parasites and their small mammalian hosts.

We studied the relationships between functional alpha and beta diversities of fleas and their small mammalian hosts in 4 biogeographic realms (the Afrotropics, the Nearctic, the Neotropics and the Palearctic), considering 3 components of alpha diversity (functional richness, divergence and regularity). We asked whether (a) flea alpha and beta diversities are driven by host alpha and beta diversities; (b) the variation in the off-host environment affects variation in flea alpha and beta diversities; and (c) the pattern of the relationship between flea and host alpha or beta diversities differs between geographic realms. We analysed alpha diversity using modified phylogenetic generalized least squares and beta diversity using modified phylogenetic generalized dissimilarity modelling. In all realms, flea functional richness and regularity increased with an increase in host functional richness and regularity, respectively, whereas flea functional divergence correlated positively with host functional divergence in the Nearctic only. Environmental effects on the components of flea alpha diversity were found only in the Holarctic realms. Host functional beta diversity was invariantly the best predictor of flea functional beta diversity in all realms, whereas the effects of environmental variables on flea functional beta diversity were much weaker and differed between realms. We conclude that flea functional diversity is mostly driven by host functional diversity, whereas the environmental effects on flea functional diversity vary (a) geographically and (b) between components of functional alpha diversity.

Essential oil pharmaceuticals for killing ectoparasites on dogs.

BACKGROUND: External parasites, particularly ticks and fleas, are among the most common problems affecting dogs. Chemical medicines are commonly used to prevent and eliminate such external parasites, but their improper use can cause adverse reactions, and the toxins they contain may remain in the environment. OBJECTIVES: The objective of this study was to investigate the in vitro efficacy of Zanthoxylum limonella, citronella, clove, peppermint, and ginger essential oils against dog ticks and fleas and to test the sensitivity of dogs' skin to these essential oils. METHODS: The five essential oils were tested for in vitro efficacy against ticks and fleas, and the two most effective essential oils were then tested on the dogs' skin. RESULTS: The results revealed that these five essential oils at 16% concentrations effectively inhibited the spawning of female engorged ticks. In addition, all five essential oils had a strong ability to kill tick larvae at concentrations of 2% upward. Furthermore, 4% concentrations of the five essential oils quickly eliminated fleas, especially clove oil, which killed 100% of fleas within 1 h. A 50%, 90%, and 99% lethal concentration (LC50, LC90, and LC99) for the essential oils on tick larvae in 24 h were found to be low values. LC50, LC90, and LC99 for the essential oils on flea in 1 h was lowest values. Clove oil at 16% concentration was the most satisfactory essential oil for application on dogs' skin, with a low percentage of adverse effects. CONCLUSIONS: This study confirmed the effectiveness of essential oils for practical use as tick and flea repellents and eliminators. Essential-oil-based pharmaceutical can replace chemical pesticides and provide benefits for both consumers and the environment.

Detection of Rickettsia raoultii in Vermipsylla alakurt-Like Fleas of Sheep in Northwestern China.

INTRODUCTION: To date, a total of 2574 validated flea species have been discovered. Vermipsyllidae is a family of fleas that comprises at least eight species. Vermipsylla is a genus of the family Vermipsyllidae within the order Siphonaptera of fleas. Here a novel Vermipsylla species was described, and rickettsial agent was also detected in it. METHODS: A total of 128 fleas were collected directly from 260 pastured sheep in China. Of these, eight representative fleas (four males and four females) were identified by key morphological features. Meanwhile, 120 flea DNAs, including six flea samples for molecular taxonomy, were subjected to Rickettsia spp. DNA detection. The molecular identity of fleas was determined by amplification and sequenmce analysis of four genetic markers (the 28S rDNA genes, the 18S rDNA genes, the mitochondrial cytochrome c oxidase subunit I and subunit II). In addition, five Rickettsia-specific gene fragments were used to identify the species of the rickettsial agents. The amplified products were sequenced and phylogenetically analyzed. RESULTS: The morphological characteristics of the flea species identified in this study were similar to Vermipsylla alakurt, but presented difference in hair number of the metepimeron, the third tergum, the genitals and the tibiae of hind leg. The 18S rDNA, 28S rDNA and COII genetic markers from fleas showed the highest identity to those of V. alakurt, shared 98.45% (954/969), 95.81% (892/931) and 85.86% (571/665) similarities, respectively. However, the COI sequence showed the highest identity to that of Dorcadia ioffi with 88.48% (576/651) similarity. Rickettsia raoutii tested positive in 14.17% (17/120) flea DNA samples. CONCLUSION: Our study reports the detection of R. raoultii in V. alakurt-like fleas infesting sheep in China.

Survey of flea infestation in cats and dogs in Western Andalusia, Spain: Seasonality and other risk factors for flea infestation.

This epidemiological survey aims to provide an update on the main flea species that parasitize domestic animals in the Western Andalusia assessing several ecological features that could be considered as possible risk factors for flea infestation. Over a 19-month period (June 2021 to January 2023), we obtained a total of 802 flea samples from 182 dogs (Canis lupus familiaris, Carnivora: Canidae, Linnaeus, 1758) and 78 cats (Felis silvestris catus, Carnivora: Felidae, Schreber, 1775). For each parasitized host, an epidemiological survey was completed, including the following information: geographical origin, age, sex, rural or urban habitat, type of animal's lifestyle (domestic or non-domestic), health status, cohabiting or not with other animals and the total number of collected fleas. The most common species was Ctenocephalides felis (Siphonaptera: Pulicidae) (Bouché, 1835) with a total of 713 specimens, which accounted for 89% of the total fleas. The second most abundant species was Pulex irritans (Siphonaptera: Pulicidae) (Linnaeus, 1758) with a total of 46 collected fleas (6% of the total). The remaining species identified were Archaeopsylla erinacei (Siphonaptera: Pulicidae) (Bouché, 1835) (25 specimens), Spilopsyllus cuniculi (Siphonaptera: Pulicidae) (Dale, 1878) (12 specimens) and Ctenocephalides canis (Siphonaptera: Pulicidae) (Curtis, 1826) (6 specimens), which accounted for 3%, 1% and 1%, respectively, of the total fleas collected. The months with the highest number of collected fleas were, in ascending order, May 2022, September 2021 and July 2021. Dogs had a greater diversity of flea species, and flea sex ratios were female biased in all identified species and among all studied hosts. Finally, we identified some potential host risk factors that promoted higher flea intensities, such as living in rural areas, or presenting other pathologies.

Host body mass, not sex, affects ectoparasite loads in yellow-necked mouse Apodemus flavicollis.

We investigated the presence and potential causes of sex bias in ectoparasite infestations in the yellow-necked mouse Apodemus flavicollis. We compared the natural tick and flea burdens of male and female mice in a temperate beech forest and assessed whether the observed differences were driven by host sex or body mass. We found that males were more heavily infested by ticks compared to female mice. However, this difference was driven by host body mass, and not sex itself. Host body mass positively correlated with flea loads, but there was no evidence of sex bias in flea abundance. In addition, the abundance of both ticks and fleas infesting yellow-necked mice changed over time, both seasonally (month to month) and annually (year to year). Our results underscore the importance of the sexual size dimorphism and the parasite taxon as the primary factors that influence the occurrence of sex-biased parasitism in small mammals.

Tick and flea infestations in captive Tapirus terrestris and Tapirus kabomani (Perissodactyla: Tapiridae) in the Brazilian Amazon.

The present study aimed to identify tick and flea species infesting captive tapirs (Tapirus terrestris and Tapirus kabomani) in the Brazilian Amazon. Ectoparasites were collected from tapirs chemically restrained in Zoo-botanical Parks, breeding facilities, conservationists and, environmental compensation areas in the states of Amapá, Amazonas, and Pará. After collection, the tick and flea specimens were placed in plastic pots containing isopropanol and identified according to dichotomous keys. Ectoparasite infestations were observed in 55% (18/33) tapirs, of which 61% (11/18) were single infestations with ticks or fleas, and 39% (7/18) were mixed infestations with different species of ticks and/or fleas. In total, 227 ticks (15 larvae, 107 nymphs, 38 females and 67 males) and 14 fleas (eight females and six males) were collected, identifying four tick species (Amblyomma cajennense sensu stricto , Amblyomma naponense, Amblyomma oblongoguttatum, and Amblyomma pacae) and three flea species and/or subspecies (Ctenocephalides felis felis, Rhopalopsyllus australis australis, and Tunga penetrans). In conclusion, infestations with Ixodidae ticks and Rhopalopsyllidae, Pulicidae and Tungidae fleas were found in captive tapirs in the Brazilian Amazon, and A. cajennense s.s. was the most frequent species. Additionally, the present study reports new associations between A. pacae and R. a. australis with T. terrestris and between A. cajennense s.s. with T. kabomani.

Compositional and phylogenetic nestedness of host assemblages exploited by generalist ectoparasites across their geographic ranges: drivers and associations with ectoparasite traits.

We investigated compositional and phylogenetic nestedness in the host assemblages of 26 host-generalist fleas across regions within the Palearctic. We asked the following questions: (i) are host assemblages exploited by a flea species compositionally or phylogenetically nested (=C-nested and P-nested, respectively) across regions?; (ii) if yes, what are the processes that generate nestedness, and does phylogenetic nestedness follow the same processes as compositional nestedness?; and (iii) are the biological traits of a flea species associated with its host assemblages' degree of nestedness? Nestedness was calculated for matrices with rows ordered either by decreasing region area (=a-matrices) or increasing distance from the centre of a flea's geographic range (d-matrices). Significant C-nestedness was found in either a- (three fleas) or d-matrices (three fleas) or both (10 fleas). Significant P-nestedness was detected in either a- (three fleas) or d-matrices (four fleas) or both (two fleas). In some but not other species, P-nestedness followed C-nestedness. The probability of C-nestedness to be significant, as well as its degree for d-matrices, was associated with a flea's morphoecological traits, whereas this was not the case for either a-matrices or the P-nestedness for either type of ordered matrices. We conclude that compositional, but not phylogenetic, nestedness is (i) generated by similar mechanisms in many flea species and (ii) may be simultaneously driven by different mechanisms in the same flea. In contrast, mechanisms promoting phylogenetic nestedness differ between flea species and seem to act separately.

Complete bacterial profile and potential pathogens of cat fleas Ctenocephalides felis.

Fleas are important ectoparasites and vectors associated with a wide range of pathogenic diseases, posing threats to public health concerns, especially cat fleas that spread worldwide. Understanding the microbial components is essential due to cat fleas are capable of transmitting pathogens to humans, causing diseases like plague and murine typhus. In the present study, metagenomic next-generation sequencing was applied to obtain the complete microbiota and related functions in the gut of Ctenocephalides felis. A total of 1,870 species was taxonomically recognized including 1,407 bacteria, 365 eukaryotes, 69 viruses, and 29 archaea. Proteobacteria was the dominant phylum among the six samples. Pathogens Rickettsia felis, Acinetobacter baumannii, Coxiella burnetii, and Anaplasma phagocytophilum were taxonomically identified and had high abundances in all samples. The resistance gene MexD was predominant in microbial communities of all cat fleas. We also performed epidemiological surveys of pathogens R. felis, A. baumannii, C. burnetii, and A. phagocytophilum among 165 cat fleas collected from seven provinces in China, while only the DNAs of R. felis (38/165, 23.03%) and C. burnetii (2/165, 1.21%) were obtained. The data provide new insight and understanding of flea intestinal microbiota and support novel information for preventing and controlling fleas and their transmitted diseases.

Host specificity of flea parasites of mammals from the Andean Biogeographic Region.

Host specificity of fleas affects their biodiversity that plays a major role in determining the potential transmission routes by pathogens through vertebrate hosts, including humans. In the Biogeographic Andean region, numerous systematic and ecological studies have been conducted, revealing a high diversity of flea taxa of mammals and the presence of pathogenic organisms transmitted by fleas; however, the degree of preference with which each flea species associates with a mammal host remains poorly understood in this region. Herein, host specificity in mammal fleas from the Andean region was analysed. We employed the number of host species for each flea species and the index of host specificity STD *. Following the literature, 144 species and 13 subspecies of fleas (31 genera and 10 families) have been described in the Andean biogeographic region; 76 taxa are endemic to this region. To carry out the analyses of host specificity, we considered 1759 records of fleas collected from 124 species and 59 genera of wild and domestic mammals, mostly rodent species (85.9%). Our results indicate that typical Andean fleas are genus or family host specific (mostly STD * less than 3.0). More diverse mammal hosts are parasitized by more diverse flea genera and families and these hosts are phylogenetically related. Otherwise, these hosts are associated with different flea lineages, suggesting the interaction of ecological and evolutionary mechanisms (host-switching, ecological adaptations and co-evolutionary alternation). The fields of disease ecology and One Health are considering the host specificity of arthropod vectors as an important point to understand the mechanisms of emergence and re-emergence of diseases. Our results allow us to estimate the risk of diseases involving fleas in the Andean region.

Predictive factors for flea occurrence in red foxes (Vulpes vulpes) from semi-arid Mediterranean environments.

The role of red fox as host for a wide range of parasites, particularly fleas and other arthropods causing vector-borne diseases, in combination with its capability to adapt to anthropized environments, makes this wild canid an epidemiologically remarkable species at the wildlife-domestic-human interface, especially in the present time of rise of emerging and re-emerging diseases. This study evaluated the prevalence and parasite intensity of fleas in 88 foxes from Murcia Region (Southeastern Spain) and determined the geographic distribution of areas with the highest potential risk of flea presence. Pulex irritans, Ctenocephalides felis, Spilopsyllus cuniculi and Nosopsyllus fasciatus were identified. The overall prevalence was 76.13%. This is the first time that N. fasciatus has been reported in foxes from Murcia Region. The predictive model established a certain pattern to determine the areas with the highest risk of acquiring fleas. Positive correlation of daily potential evapotranspiration (ET0 ) in winter and the opposite effect occurring for ET0 in summer were obtained, as well as positive correlations for mean daily temperature (Tmean ) in summer and mean precipitation (Pmean ) in winter and summer. The model was also found positively correlated in the forest habitat ecotone areas and the anthropized areas.

Fleas from the Silk Road in Central Asia: identification of Ctenocephalides canis and Ctenocephalides orientis on owned dogs in Uzbekistan using molecular identification and geometric morphometrics.

BACKGROUND: The Silk Road connected the East and West for over 1500 years. Countries in Central Asia are valuable in addressing the hypothesis that parasites on domestic animals were introduced along the Silk Road. Adult fleas are obligate parasites, having worldwide distribution. In dogs, Ctenocephalides canis, C. felis and C. orientis are the most common species identified. The distribution of the Oriental cat flea, C. orientis, is restricted to southeast Asia. The purpose of this study was to determine the diversity of dog fleas from Uzbekistan, a country in Central Asia, with particular reference to C. orientis. METHODS: Fleas were collected from 77 dogs from 5 locations in Uzbekistan. The cox1 gene sequences from Ctenocephalides spp. were compared to global collection of Ctenocephalides cox1 haplotypes. Landmark-based geometric morphometrics have been applied to the head and curvature to compare C. canis and C. canis using canonical variate analysis and discriminant function analysis. RESULTS: Overall, 199 fleas were collected and identified as C. canis (n = 115, 58%), C. orientis (n = 53, 27%) and Pulex irritans (n = 22, 11%). None of the fleas were C. felis. All Ctenocephalides spp. fleas were subject to cox1 amplification and 95% (166/175) yielded DNA sequence. There were 25 cox1 haplotypes; 14 (22/25, 88%) were C. canis cox1 haplotypes and 3 (3/25, 12%) were C. orientis cox1 haplotypes. Molecular analysis confirmed the absence of C. felis. Four (4/22) and one (1/3) cox1 haplotypes were identical to cox1 haplotypes belonging to C. canis and C. orientis cox1 haplotypes identified elsewhere, respectively. Overall morphometric analysis confirmed significant differences between the head shape of C. canis and C. orientis and improved four-fivefold the species identification compared to traditional morphological key. CONCLUSION: We report for the first time the presence of C. orientis in Uzbekistan. Differentiation of C. orientis from C. canis and C. felis remains difficult in regions where these species coexist. Studies in Central and Southeast Asia should confirm species identity using cox1 locus to enable retracing of the distribution of the Ctenocephalides in Asia. The presence of C. orientis suggests that this species may have been introduced from the east along the ancient Silk Road.

Prevalence of selected pathogens in ectoparasites from free-roaming domestic cats in the southern Great Plains of the United States.

Rickettsia sp. and Bartonella sp. were detected in ectoparasites of free-roaming domestic cats (Felis catus) from a trap-neuter-release program in central Oklahoma during January and February 2020. We used polymerase chain reaction and sequencing to identify fleas containing DNA of five different pathogens: Rickettsia felis (84%), 'Candidatus R. andeanae' (8%), Bartonella henselae (32%), Bartonella clarridgeiae (36%), and Bartonella sp. (8%). Co-infections with R. felis with three Bartonella species were identified. One tick was positive for R. felis, one flea was positive for 'Candidatus R. andeanae' and one ear mite was positive for a Bartonella species. These results highlight the need for more focus on free-roaming domestic cats and their ectoparasites in the Great Plains region.

Mitochondrial phylogenomics provides insights into the taxonomy and phylogeny of fleas.

BACKGROUND: Fleas (Insecta: Siphonaptera) are obligatory hematophagous ectoparasites of humans and animals and serve as vectors of many disease-causing agents. Despite past and current research efforts on fleas due to their medical and veterinary importance, correct identification and robust phylogenetic analysis of these ectoparasites have often proved challenging. METHODS: We decoded the complete mitochondrial (mt) genome of the human flea Pulex irritans and nearly complete mt genome of the dog flea Ctenocephalides canis, and subsequently used this information to reconstruct the phylogeny of fleas among Endopterygota insects. RESULTS: The complete mt genome of P. irritans was 20,337 bp, whereas the clearly sequenced coding region of the C. canis mt genome was 15,609 bp. Both mt genomes were found to contain 37 genes, including 13 protein-coding genes, 22 transfer RNA genes and two ribosomal RNA genes. The coding region of the C. canis mt genome was only 93.5% identical to that of the cat flea C. felis, unequivocally confirming that they are distinct species. Our phylogenomic analyses of the mt genomes showed a sister relationship between the order Siphonaptera and orders Diptera + Mecoptera + Megaloptera + Neuroptera and positively support the hypothesis that the fleas in the order Siphonaptera are monophyletic. CONCLUSIONS: Our results demonstrate that the mt genomes of P. irritans and C. canis are different. The phylogenetic tree shows that fleas are monophyletic and strongly support an order-level objective. These mt genomes provide novel molecular markers for studying the taxonomy and phylogeny of fleas in the future.