Search Results (33)

The ionotropic γ-aminobutyric acid (GABA) receptor (GABAR) is a key target for the development of antiparasitic agents, particularly against ectoparasites, such as fleas and ticks. Binding stability and selectivity of sarolaner enantiomers for Ctenocephalides felis RDL receptors (RDLR) were investigated in the current study. Wild-type (WT) C. felis RDLR and its A285S mutant were constructed using homology-based, fragment-based threading and AI-driven approaches, of which, SWISS-MODEL generated the most reliable structures. Molecular docking showed that the sarolaner S-enantiomer had higher binding affinity for both receptors than the R-enantiomer, primarily due to hydrogen bonding with Ile256, π–π stacking with Phe326, and hydrophobic interactions with Ile267 and Ile268. Molecular dynamics simulations confirmed the binding stability of the S-enantiomer-receptor complex in which key residues maintained interactions throughout the trajectories. Binding free energy analysis supported these results and highlighted the role of nonpolar interactions in binding stability. The A285S mutation had minimal impact on the binding pocket, and the S-enantiomer remained selective for and bound to the mutant receptor. Insights into the insecticidal mechanism of sarolaner enantiomers are given, and the current findings may inform the development of veterinary drugs from novel isoxazoline-based NAMs targeting insect GABARs. Full article

Bartonella henselae is an important zoonotic pathogen with a wide range of clinical manifestations in humans. Despite advances in understanding its pathogenesis, there is no broadly applicable laboratory animal model for bartonellosis. This study aimed to assess the potential utility of an experimental model of chronic B. henselae infection using ferrets, a species previously utilized in various human pathogen studies. Six ferrets (n = 6) were divided into three groups: a control group (n = 2), a low-dose infection group (n = 2), and a high-dose infection group (n = 2). The two infection groups were inoculated intradermally with 10⁵ (low dose) and 10⁹ (high dose) CFU/mL B. henselae, respectively. Clinical signs, serological responses, and bacteriological findings were monitored over seven weeks; ferrets were then euthanized and tissues were examined histologically. Only minimal or transient systemic clinical signs and laboratory abnormalities developed in B. henselae inoculated ferrets. The high-dose group seroconverted to B. henselae antigen within two weeks, maintaining elevated titers throughout the study. Histopathological examination revealed that four B. henselae-infected ferrets had notable microscopic inflammatory lesions in the liver parenchyma (3/4), heart (1/4), and brain (1/4); similar lesions were not observed in the tissues of the two control ferrets. Despite the presence of microscopic lesions and seroconversion in the high-dose group, bacteremia was not documented and B. henselae DNA was not successfully amplified by quantitative PCR from lesional organs. This pilot study demonstrated that ferrets may serve as a promising model for investigating B. henselae pathogenesis, diagnosis, treatment, and prevention. Full article

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. Full article

Flea-borne typhus (FBT), due to Rickettsia typhi and R. felis, is an infection typically causing fever, headache, rash, hepatitis, and thrombocytopenia. About one quarter of patients suffer pulmonary, neurologic, hematologic, renal, hepatic, cardiac, ocular or other complications. In the 21st century, the incidence of FBT has increased in both Texas and California compared to the 1990s. In this paper, county-level epidemiological data for the number of cases of FBT occurring in Texas for two decades, 1990–1999 and 2010–2019, were compared with respect to county of residence, urbanization, and climatic region. Human population growth in Texas has promoted FBT by increased urbanization and the abundance of pet dogs and cats, stray/feral dogs and cats, and opossums. Increasing temperatures in Texas in the new millennium have increased the flea-borne transmission of FBT by promoting host infestation and flea feeding and defecation, accelerating the flea life cycle, and increasing rickettsial replication within the flea. Increased numbers of opossums and stray cats and dogs in the urban/suburban landscape have increased the risk of flea transfer to humans and their pets. Full article

This study reinforces the value of a One Health approach to infectious disease outbreak investigations. After the onset of neuropsychiatric symptoms in their son, our investigation focused on a family composed of a mother, father, two daughters, the son, two dogs, and a rabbit, all with exposures to vectors (fleas and ticks), rescued dogs, and other animals. Between 2020 and 2022, all family members experienced illnesses that included neurological symptoms. Prolonged menorrhagia (130d) in the youngest daughter ultimately resolved following antibiotic administration. One dog was diagnosed with a splenic hematoma and months later spinal histiocytic sarcoma. The father, both daughters, and one dog were seroreactive to multiple Bartonella spp. antigens, whereas the mother and son were not seroreactive. Bartonella quintana DNA was amplified from specimens obtained from all family members. Based upon DNA sequencing, infection with B. quintana was confirmed for the mother and both pet dogs. Bartonella henselae DNA was amplified and sequenced from the youngest daughter, the son, and one dog (co-infected with B. quintana), and from Ctenocephalides felis collected from their pet rabbit. All five family members and one dog were infected with Babesia divergens-like MO-1. Both parents were co-infected with Babesia microti. Droplet digital PCR supported potential infection with a Borrelia species in three family members. This study provided additional case-based evidence supporting the role of stealth Babesia, Bartonella, and Borrelia pathogens as a cause or cofactor in neurological and neuropsychiatric symptoms. We conclude that a One Health investigation approach, particularly for stealth vector borne pathogens such as Babesia, Bartonella, and Borrelia spp., will enhance clinical and epidemiological understanding of these organisms for animal and human health. During outbreak investigations it is critical to document travel and vector exposure histories, symptoms, and pathology in pets and human patients, contact with rescued, wild, or feral animals and perform diagnostic testing that includes family members, pets, and vectors. Full article

The Trypanosomatidae family encompasses around 24 genera of unicellular protozoans, many of which are transmitted by various hematophagous arthropods, particularly members of the Orders Diptera and Hemiptera. Fleas and ticks—an understudied group of ectoparasites—have been shown to be hosts of a wide and crescent variety of trypanosomatid species. Further, fleas and ticks of companion animals have been particularly neglected in trypanosomatid surveillance despite the proximity to human populations and the anthropophagous habits of many of these arthropods, which can potentially act as vectors of zoonotic trypanosomatids. We aimed to identify the presence, characterize the species, and establish the prevalence of Kinetoplastids, including members of the Trypanosomatidae family, in ectoparasites collected from dogs and humans from Mexico. A total of 537 ectoparasite specimens belonging to six ectoparasite taxa (Amblyomma mixtum, A. tenellum, Ctenocephalides felis felis, Pulex simulans, Rhipicephalus linnaei, and Rh. sanguineus s.s.) were collected from 15 States of Mexico. An 800 bp fragment of the 18S-rDNA gene from kinetoplastids was amplified and sequenced. The presence of two agents (Trypanosoma caninum and Parabodo sp.) was detected in R. linnaei ticks and one (Blechomonas lauriereadi) in the cat flea Ct. felis felis. This is the first record of genetic material of kinetoplastid species in ectoparasites from dogs and humans in Mexico. Full article

Fleas (Siphonaptera) are ectoparasitic hematophagous insects responsible for causing bites and itchy skin conditions in both humans and animals. Furthermore, they can act as vectors of different pathogens of a wide variety of diseases worldwide, including bartonellosis, rickettsiosis, and bubonic plague. Accurate identification of fleas is necessary for the study of their epidemiology, prevention, and control. In addition to traditional morphological classification approaches and molecular biology techniques, geometric morphometrics is increasingly proving to be a useful complementary tool for discriminating between Siphonaptera taxa. With the objective of determining the capacity of this technique to identify and differentiate synanthropic fleas, a principal component analysis was carried out on populations of Ctenocephalides felis, Pulex irritans, and Archaeopsylla erinacei collected in distinct regions of Andalusia (Spain). The analysis carried out on 81 male and female specimens revealed factorial maps that allowed the differentiation of the populations under study, with only partial overlaps that did not prevent their correct identification. Global size differences were also detected, with a slightly larger size in P. irritans males and a bigger size in A. erinacei females. Therefore, the present study emphasizes the role of geometric morphometrics as a useful complementary technique in taxonomic studies of arthropods, especially in the case of flea specimens lacking representative morphological features. Full article

Bartonella henselae is associated with numerous clinical syndromes in people. Cats are the definitive hosts for B. henselae, develop high levels of bacteremia, and are associated with human infections, particularly in the presence of Ctenocephalides felis. Several antibiotic protocols used for the treatment of B. henselae infection in cats have failed to clear bacteremia. The purpose of this study was to assess the safety and efficacy of a high-dose pradofloxacin protocol to eliminate B. henselae bacteremia. Bartonella henselae infection was initiated in 8 cats by intravenous inoculation of infected feline blood and then pradofloxacin was administered at 7.5 mg/kg, PO, twice daily for 28 days, starting 12 weeks after inoculation. Complete blood cell counts were performed prior to pradofloxacin administration and then every 2 weeks for 10 weeks. Bartonella PCR assay was performed prior to pradofloxacin administration and approximately every 2 weeks for 10 weeks and then weekly for 4 weeks. Methylprednisolone acetate (5 mg/kg) was administered by intramuscular injection to all cats on week 10. The cats remained normal and none developed a hematocrit, platelet count, lymphocyte count, or neutrophil count outside of the normal reference ranges. In the one month prior to pradofloxacin administration, all cats were PCR-positive for Bartonella DNA on at least two of four sample dates; after pradofloxacin administration, all cats were negative for B. henselae DNA in blood on all nine sample dates. The protocol appears to be safe and failure to amplify B. henselae DNA from the blood after the administration of pradofloxacin and one dose of methylprednisolone acetate suggests either an antibiotic effect or the organism was cleared spontaneously. Full article

Mycoplasma spp. pathogens frequently cause chronic and acute diseases in cats. The aim of the present study was to investigate the presence and genetic diversity of Mycoplasma spp. in cats and their ectoparasites using PCR and sequence analysis of the 16S rRNA gene. Blood samples were collected from 541 domestic and stray cats in Lithuania. Ectoparasites (153 fleas and 321 ticks) were collected from owned domestic cats that live both outdoors and indoors. Mycoplasma spp. were detected in 7.2% of cat blood samples and 4.4% of Ctenocephalides felis fleas. The sequence analysis revealed the presence of Mycoplasma haemofelis in 1.1% of cats and ‘Candidatus Mycoplasma haematominutum’ in 4.8% of cats. Ct. felis fleas harboured M. haemofelis. To the best of the authors’ knowledge, this is the first report on the prevalence and molecular characterisation of Mycoplasma bacteria in cats in Lithuania and cat fleas in the Baltic States. Full article

Zoonotic parasitic diseases in dogs are particularly concerning in regions with low human development indices due to inadequate sanitary services and insufficient environmental and health education. This study aimed to assess the parasitological status of dogs living in households and evaluate their owners’ knowledge about zoonoses. A total of 183 dogs from Rolim de Moura, Rondônia State, were screened for the presence of ectoparasites, and 163 fecal samples were collected for analysis. The results showed that 74.23% (112/163) of the animals had at least one species of endoparasite. The most identified pathogens were Ancylostoma spp. (68.71%, 112/163), Trichuris vulpis (11.66%, 19/163), Toxocara canis (6.75%, 11/163), Cystoisospora canis (4.91%, 8/163), Dipylidium caninum (1.23%, 2/163), and Hammondia/Neospora (0.61%, 1/163). Ectoparasites were observed in 43.17% (79/183) of the evaluated animals, with Rhipicephalus sanguineus found in 31.15% (57/183) and Ctenocephalides felis felis in 20.77% (38/183). Only 11.48% (7/61) of the owners were familiar with the term “Zoonoses.” However, a significant majority (83.61%, 51/61) believed that dogs can transmit diseases to humans. Our findings highlight the prevalence of parasites in the studied area and associated risk factors, underscoring the urgent need for educational interventions to raise awareness about these diseases and their risks to human health. Full article

Murine typhus is an acute febrile illness caused by Rickettsia typhi, an obligately intracellular Gram-negative coccobacillus. Rats (Rattus species) and their fleas (Xenopsylla cheopis) serve as the reservoir and vector of R. typhi, respectively. Humans become infected when R. typhi-infected flea feces are rubbed into flea bite wounds or onto mucous membranes. The disease is endemic throughout much of the world, especially in tropical and subtropical seaboard regions where rats are common. Murine typhus is reemerging as an important cause of febrile illness in Texas and Southern California, where an alternate transmission cycle likely involves opossums (Didelphis virginiana) and cat fleas (Ctenocephalides felis). Although primarily an undifferentiated febrile illness, a range of neurologic manifestations may occur, especially when treatment is delayed. Serology is the mainstay of diagnostic testing, but confirmation usually requires demonstrating seroconversion or a fourfold increase in antibody titer from acute- and convalescent-phase sera (antibodies are seldom detectable in the first week of illness). Thus, early empiric treatment with doxycycline, the drug of choice, is imperative. The purpose of this review is to highlight murine typhus as an important emerging and reemerging infectious disease, review its neurologic manifestations, and discuss areas in need of further study. Full article

This overview of extant Siphonaptera lists 19 families with major hosts and their general distribution, estimated numbers of genera, species, and subspecies, with a brief taxonomic and phylogenetic review. With around 10 new species described annually, extant flea fauna comprises an estimated 249 genera, 2215 species, and 714 subspecies globally, mostly mammal parasites, but 5% of species are on birds. Host specificity varies from euryxenous (i.e., infesting two or more host orders) (e.g., cat fleas, Ctenocephalides felis felis) to monoxenous (e.g., rabbit fleas, Spilopsyllus cuniculi). The largest family is the paraphyletic Hystrichopsyllidae, making up a third of all flea species. The largest monophyletic family, Ceratophyllidae (rodent and bird fleas), comprises another 20% and has dispersed to every continent, including Antarctica. Fleas descend from scorpionflies (Mecoptera), possibly snow scorpionflies (Boreidae) or Nannochoristidae, and even giant fossils found from the Mesozoic could be Siphonaptera. The diversification of fleas shows evidence of taxon cycles. “Relict” families, such as helmet fleas (Stephanocircidae), have a disjunct distribution reflecting the breakup of Gondwanaland 70 million years ago. “Niche specialists” include nest fleas (Anomiopsyllus), bat fleas (Ischnopsyllidae), and burrowing fleas, such as chigoes (Tungidae). By contrast, Ceratophyllidae fleas could be considered “great speciators”. Cat fleas and several other synanthropic flea species are invasive “supertramps”. Although those species are intensively studied, many flea species and their hosts require urgent surveys and conservation. Full article

MALDI-TOF is now considered a relevant tool for the identification of arthropods, including lice and fleas. However, the duration and conditions of storage, such as in ethanol, which is frequently used to preserve these ectoparasites, could impede their classification. The purpose of the present study was to assess the stability of MS profiles from Pediculus humanus corporis lice and Ctenocephalides felis fleas preserved in alcohol from one to four years and kinetically submitted to MALDI-TOF MS. A total of 469 cephalothoraxes from lice (n = 170) and fleas (n = 299) were tested. The reproducibility of the MS profiles was estimated based on the log score values (LSVs) obtained for query profiles compared to the reference profiles included in the MS database. Only MS spectra from P. humanus corporis and C. felis stored in alcohol for less than one year were included in the reference MS database. Approximately 75% of MS spectra from lice (75.2%, 94/125) and fleas (74.4%, 122/164) specimens stored in alcohol for 12 to 48 months, queried against the reference MS database, obtained relevant identification. An accurate analysis revealed a significant decrease in the proportion of identification for both species stored for more than 22 months in alcohol. It was hypothesized that incomplete drying was responsible for MS spectra variations. Then, 45 lice and 60 fleas were subjected to longer drying periods from 12 to 24 h. The increase in the drying period improved the proportion of relevant identification for lice (95%) and fleas (80%). This study highlighted that a correct rate of identification by MS could be obtained for lice and fleas preserved in alcohol for up to four years on the condition that the drying period was sufficiently long for accurate identification. Full article

Fleas are ectoparasites affecting many animal species but reports in captive nonhuman primates are rare and mainly concern pet monkeys. Moreover, to the authors’ knowledge, a detailed report on marmosets is not known at present. This case describes the clinical signs, diagnosis, treatment and follow-up of a flea infestation by Ctenocephalides felis in a captive colony of common marmosets. Fleas, flea feces and skin lesions were identified on two animals during annual health screening. Subsequently, the entire colony was examined, and nearly half of the colony showed signs of infestation. Consequently, treatment was initiated for the entire colony and the environment. Animals received two topical administrations of imidacloprid (5 mg for animals <200 g and 10 mg for animals weighing >200 g) three weeks apart, and their enclosures were decontaminated using vaporizers containing permethrin, piperonyl butoxide, and pyriproxyfen. Subsequently, skin lesions were resolved and no evidence of fleas were noticed. No side effects of the treatment were observed. Stray cats were identified as the source of the infestation. Their access to the animal-related rooms was stopped. No reinfestation has been reported for 3 years. The topical application of imidacloprid appeared effective with no adverse events occurring, so may be appropriate for use in other non-human primates. Full article

Fleas are obligatory blood-sucking ectoparasites of medical and veterinary importance. The identification of fleas and associated flea-borne microorganisms, therefore, plays an important role in controlling and managing these vectors. Recently, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been reported as an innovative and effective approach to the identification of arthropods, including fleas. This study aims to use this technology to identify ethanol-preserved fleas collected in Vietnam and to use molecular biology to search for microorganisms associated with these fleas. A total of 502 fleas were collected from wild and domestic animals in four provinces in Vietnam. Morphological identification led to the recognition of five flea species, namely Xenopsylla cheopis, Xenopsylla astia, Pulex irritans, Ctenocephalides canis, and Ctenocephalides felis. The cephalothoraxes of 300 individual, randomly selected fleas were tested using MALDI-TOF MS and molecular analysis for the identification and detection of microorganisms. A total of 257/300 (85.7%) of the obtained spectra from the cephalothoraxes of each species were of good enough quality to be used for our analyses. Our laboratory MALDI-TOF MS reference database was upgraded with spectra achieved from five randomly selected fleas for every species of Ctenocephalides canis and Ctenocephalides felis. The remaining spectra were then queried against the upgraded MALDI-TOF MS database, which showed 100% correspondence between morphology and MALDI-TOF MS identification for two flea species (Ctenocephalides canis and Ctenocephalides felis). The MS spectra of the remaining species (three P. irritans, five X. astia, and two X. cheopis) were visually generated low-intensity MS profiles with high background noise that could not be used to update our database. Bartonella and Wolbachia spp. were detected in 300 fleas from Vietnam using PCR and sequencing with primers derived from the gltA gene for Bartonella and the 16S rRNA gene for Wolbachia, including 3 Bartonella clarridgeiae (1%), 3 Bartonella rochalimae (1%), 1 Bartonella coopersplainsensis (0.3%), and 174 Wolbachia spp. endosymbionts (58%). Full article