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).

New records of pathogenic bacteria in different species of fleas collected from domestic and peridomestic animals in Spain. A potential zoonotic threat?

Climate change is causing many vectors of infectious diseases to expand their geographic distribution as well as the pathogens they transmit are also conditioned by temperature for their multiplication. Within this context, it is worth highlighting the significant role that fleas can play as vectors of important pathogenic bacteria. For this purpose, our efforts focused on detecting and identifying a total of 9 bacterial genera (Rickettsia sp.; Bartonella sp.; Yersinia sp.; Wolbachia sp., Mycobacterium sp., Leishmania sp., Borrelia sp., Francisella sp. and Coxiella sp.) within fleas isolated from domestic and peridomestic animals in the southwestern region of Spain (Andalusia). Over a 19-months period, we obtained flea samples from dogs, cats and hedgehogs. A total of 812 fleas was collected for this study. Five different species were morphologically identified, including C. felis, C. canis, S. cuniculi, P. irritans, and A. erinacei. Wolbachia sp. was detected in all five species identified in our study which a total prevalence of 86%. Within Rickettsia genus, two different species, R. felis and R. asembonensis were mainly identified in C. felis and A. erinacei, respectively. On the other hand, our results revealed a total of 131 fleas testing positive for the presence of Bartonella sp., representing a prevalence rate of 16% for this genus identifying two species B. henselae and B. clarridgeiae. Lastly, both Y. pestis and L. infantum were detected in DNA of P. irritans and C. felis, respectively isolated from dogs. With these data we update the list of bacterial zoonotic agents found in fleas in Spain, emphasizing the need to continue conducting future experimental studies to assess and confirm the potential vectorial role of certain synanthropic fleas.

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.

Vector biology of the cat flea Ctenocephalides felis.

Ctenocephalides felis, the cat flea, is among the most prevalent and widely dispersed vectors worldwide. Unfortunately, research on C. felis and associated pathogens (Bartonella and Rickettsia spp.) lags behind that of other vectors and vector-borne pathogens. Therefore, we aimed to review fundamental aspects of C. felis as a vector (behavior, epidemiology, phylogenetics, immunology, and microbiome composition) with an emphasis on key techniques and research avenues employed in other vector species. Future laboratory C. felis experimental infections with Bartonella, Rickettsia, and Wolbachia species/strains should examine the vector-pathogen interface utilizing contemporary visualization, transcriptomic, and gene-editing techniques. Further environmental sampling will inform the range and prevalence of C. felis and associated pathogens, improving the accuracy of vector and pathogen modeling to improve infection/infestation risk assessment and diagnostic recommendations.

Complex effects of testosterone level on ectoparasite load in a ground squirrel: an experimental test for the immunocompetence handicap hypothesis.

BACKGROUND: The immunocompetence handicap hypothesis suggests that males with a higher testosterone level should be better at developing male secondary traits, but at a cost of suppressed immune performance. As a result, we should expect that males with an increased testosterone level also possess a higher parasite load. However, previous empirical studies aimed to test this prediction have generated mixed results. Meanwhile, the effect of testosterone level on parasite load in female hosts remains poorly known. METHODS: In this study, we tested this prediction by manipulating testosterone level in Daurian ground squirrels (Spermophilus dauricus), a medium-sized rodent widely distributed in northeast Asia. S. dauricus is an important host of ticks and fleas and often viewed as a considerable reservoir of plague. Live-trapped S. dauricus were injected with either tea oil (control group) or testosterone (treatment group) and then released. A total of 10 days later, the rodents were recaptured and checked for ectoparasites. Fecal samples were also collected to measure testosterone level of each individual. RESULTS: We found that testosterone manipulation and sex of hosts interacted to affect tick load. At the end of the experiment, male squirrels subjected to testosterone implantation had an averagely higher tick load than males from the control group. However, this pattern was not found in females. Moreover, testosterone manipulation did not significantly affect flea load in S. dauricus. CONCLUSIONS: Our results only lent limited support for the immunocompetence handicap hypothesis, suggesting that the role of testosterone on regulating parasite load is relatively complex, and may largely depend on parasite type and gender of hosts.

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.

Nosopsyllus (Nosopsyllus) spiniformis n. sp., a new ceratophyllid flea (Siphonaptera, Ceratophyllidae, Ceratophyllinae) from Trkiye.

A new ceratophyllid flea species, Nosopsyllus (Nosopsyllus) spiniformis n. sp., is described and illustrated. Specimens were collected from Gnthers vole, Microtus guentheri (Danford & Alston), Schidlovskys vole, Microtus schidlovskii Argyropulo, in Adana province, and stone marten, Martes foina (Erxleben), in Tokat province. With the new species, the number of Nosopsyllus taxa known from Trkiye now tallies 11 (six species and five subspecies).

Socio-ecological risk factors associated with human flea infestations of rural household in plague-endemic areas of Madagascar.

Plague is a flea-borne fatal disease caused by the bacterium Yersinia pestis, which persists in rural Madagascar. Although fleas parasitizing rats are considered the primary vectors of Y. pestis, the human flea, Pulex irritans, is abundant in human habitations in Madagascar, and has been found naturally infected by the plague bacterium during outbreaks. While P. irritans may therefore play a role in plague transmission if present in plague endemic areas, the factors associated with infestation and human exposure within such regions are little explored. To determine the socio-ecological risk factors associated with P. irritans infestation in rural households in plague-endemic areas of Madagascar, we used a mixed-methods approach, integrating results from P. irritans sampling, a household survey instrument, and an observational checklist. Using previously published vectorial capacity data, the minimal P. irritans index required for interhuman bubonic plague transmission was modeled to determine whether household infestations were enough to pose a plague transmission risk. Socio-ecological risk factors associated with a high P. irritans index were then identified for enrolled households using generalized linear models. Household flea abundance was also modeled using the same set of predictors. A high P. irritans index occurred in approximately one third of households and was primarily associated with having a traditional dirt floor covered with a plant fiber mat. Interventions targeting home improvement and livestock housing management may alleviate flea abundance and plague risk in rural villages experiencing high P. irritans infestation. As plague-control resources are limited in developing countries such as Madagascar, identifying the household parameters and human behaviors favoring flea abundance, such as those identified in this study, are key to developing preventive measures that can be implemented at the community level.

Cat Flea Coinfection with Rickettsia felis and Rickettsia typhi.

Purpose: Flea-borne rickettsioses, collectively referred to as a term for etiological agents Rickettsia felis, Rickettsia typhi, and RFLOs (R. felis-like organisms), has become a public health concern around the world, specifically in the United States. Due to a shared arthropod vector (the cat flea) and clinical signs, discriminating between Rickettsia species has proven difficult. While the effects of microbial coinfections in the vector can result in antagonistic or synergistic interrelationships, subsequently altering potential human exposure and disease, the impact of bacterial interactions within flea populations remains poorly defined. Methods: In this study, in vitro and in vivo systems were utilized to assess rickettsial interactions in arthropods. Results: Coinfection of both R. felis and R. typhi within a tick-derived cell line indicated that the two species could infect the same cell, but distinct growth kinetics led to reduced R. felis growth over time, regardless of infection order. Sequential flea coinfections revealed the vector could acquire both Rickettsia spp. and sustain coinfection for up to 2 weeks, but rickettsial loads in coinfected fleas and feces were altered during coinfection. Conclusion: Altered rickettsial loads during coinfection suggest R. felis and R. typhi interactions may enhance the transmission potential of either agent. Thus, this study provides a functional foundation to disentangle transmission events propelled by complex interspecies relationships during vector coinfections.

Identification of tungiasis infection hotspots with a low-cost, high-throughput method for extracting Tunga penetrans (Siphonaptera) off-host stages from soil samples-An observational study.

BACKGROUND: The sand flea, Tunga penetrans, is the cause of a severely neglected parasitic skin disease (tungiasis) in the tropics and has received little attention from entomologists to understand its transmission ecology. Like all fleas, T. penetrans has environmental off-host stages presenting a constant source of reinfection. We adapted the Berlese-Tullgren funnel method using heat from light bulbs to extract off-host stages from soil samples to identify the major development sites within rural households in Kenya and Uganda. METHODS AND FINDINGS: Simple, low-cost units of multiple funnels were designed to allow the extraction of >60 soil samples in parallel. We calibrated the method by investigating the impact of different bulb wattage and extraction time on resulting abundance and quality of off-host stages. A cross-sectional field survey was conducted in 49 tungiasis affected households. A total of 238 soil samples from indoor and outdoor living spaces were collected and extracted. Associations between environmental factors, household member infection status and the presence and abundance of off-host stages in the soil samples were explored using generalized models. The impact of heat (bulb wattage) and time (hours) on the efficiency of extraction was demonstrated and, through a stepwise approach, standard operating conditions defined that consistently resulted in the recovery of 75% (95% CI 63-85%) of all present off-host stages from any given soil sample. To extract off-host stages alive, potentially for consecutive laboratory bioassays, a low wattage (15-25 W) and short extraction time (4 h) will be required. The odds of finding off-host stages in indoor samples were 3.7-fold higher than in outdoor samples (95% CI 1.8-7.7). For every one larva outdoors, four (95% CI 1.3-12.7) larvae were found indoors. We collected 67% of all off-host specimen from indoor sleeping locations and the presence of off-host stages in these locations was strongly associated with an infected person sleeping in the room (OR 10.5 95% CI 3.6-28.4). CONCLUSION: The indoor sleeping areas are the transmission hotspots for tungiasis in rural homes in Kenya and Uganda and can be targeted for disease control and prevention measures. The soil extraction methods can be used as a simple tool for monitoring direct impact of such interventions.

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.

Evidence of castrations in fleas of small mammals (Chiroptera and Rodentia) from Argentina.

Morphological abnormalities in fleas seem to be common in nature and are under reported in Argentina. In this note, we describe anomalies in two males and one female of Alectopsylla unisetosa Mahnert (Ischnopsyllidae) and one of Polygenis sp. (Rhopalopsyllidae) male collected from small mammals in the provinces of Neuquen and Salta, Argentina. In all specimens, the anomalies were observed at the level of the genitalia recognized as partial castration. The structures mainly affected were the modified abdominal segments, the aedeagus (in male), and the spermatheca (in female). The present communication is the first one devoted exclusively to teratogenous fleas in Argentina.

Morpho-molecular investigation of ectoparasitic infestation of companion animals in Sylhet city, Bangladesh.

Ticks (Ixodida) and Fleas (Siphonaptera) are considered among the most important arthropod of public health concern due to their ability to transmit vector-borne pathogens to humans. By sharing a common environment, vector-borne diseases constituted major setbacks to the development of a pet population in Bangladesh. This study aimed to determine companion animal-associated ticks and fleas based on morpho-molecular approaches. Between December 2021 and May 2022, 74 animals (62 cats and 12 dogs) were examined, of which 17 (27.4%) cats and 9 (75.0%) dogs had ectoparasitic infestations, with 35.1% overall prevalence. Morphometrical examination showed the ectoparasites in these animals were Ctenocephalides spp. (flea) and Riphicephalus spp. (tick). Genetic analysis using the mitochondrial markers i.e. Cytochrome c oxidase subunit 1 (cox1) revealed the presence of two flea species i.e., Ctenocephalides canis, Ctenocephalides felis, and one tick species Rhipicephalus sanguineus. Interviews of animal owners indicate that 35.14% of them had no concern about ectoparasitic infestation or ectoparasites-borne diseases. Our results indicated that fleas and ticks were the most common ectoparasites in companion animals of this area. The zoonotic nature of some ectoparasites can be regarded as a public health alert. The findings will assist epidemiologists and policymakers in offering customized guidance for upcoming monitoring and preventive tactics in this area.

Molecular detection and identification of hemotropic Mycoplasma species in dogs and their ectoparasites in Iran.

Hemotropic Mycoplasma species are vector-borne bacteria that attach and grow on the surface of erythrocytes in various mammals, yet reports of canine hemoplasmosis in Iran are scarce. The aim of this study was molecular detection and identification of hemoplasmas in the blood of dogs (n = 370) from five provinces of Iran and ectoparasites infesting them including Ctenocephalides canis and Pulex irritans fleas, Rhipicephalus sanguineus sensu lato ticks, Heterodoxus spiniger lice and Hippobosca longipennis keds. Hemotropic Mycoplasma spp. pathogens were detected using genus-specific conventional PCRs, and subsequently identified using species-specific PCRs for Mycoplasma haemocanis (Mhc), and Candidatus Mycoplasma haematoparvum (CMhp). Sanger sequencing was then performed to confirm the species. Correlation of infection and risk factors (geographical area, keeping condition, body condition, sex, age, ectoparasite infestation) were analyzed. In total, 210 dogs (56.7%) were tested PCR-positive for hemotropic Mycoplasma spp. Species-specific PCR and sequencing revealed infection with Mhc in 17.8%, with CMhp in 7.02% and co-infection in 31.9% of dogs. Flea infestation, poor body condition, and being older than 3-years-old correlated with hemoplasmosis. In ectoparasites, DNA of hemoplasmas were detected only in fleas i.e. Mhc in P. irritans, CMhp in P. irritans and C. canis, and co-infection in C. canis. To our knowledge, this is the first large-scale molecular epidemiology study of canine hemoplasmosis in Iran. Considering the high prevalence of canine hemoplasmosis all over the country including potentially zoonotic CMhp, effective ectoparasite control strategies, regular examination of dogs, successful chemoprophylaxis and public awareness strategies are advocated.

A nowcast model to predict outdoor flea activity in real time for the contiguous United States.

BACKGROUND: The cat flea (Ctenocephalides felis), a parasite commonly found on both dogs and cats, is a competent vector for several zoonotic pathogens, including Dipylidium caninum (tapeworms), Bartonella henselae (responsible for cat scratch disease) and Rickettsia felis (responsible for flea-borne spotted fever). Veterinarians recommend that both cats and dogs be routinely treated with medications to prevent flea infestation. Nevertheless, surveys suggest that nearly one third of pet owners do not routinely administer appropriate preventatives. METHODS: A mathematical model based on weighted averaging over time is developed to predict outdoor flea activity from weather conditions for the contiguous United States. This 'nowcast' model can be updated in real time as weather conditions change and serves as an important tool for educating pet owners about the risks of flea-borne disease. We validate our model using Google Trends data for searches for the term 'fleas.' This Google Trends data serve as a proxy for true flea activity, as validating the model by collecting fleas over the entire USA is prohibitively costly and time-consuming. RESULTS: The average correlation (r) between the nowcast outdoor flea activity predictions and the Google Trends data was moderate: 0.65, 0.70, 0.66, 0.71 and 0.63 for 2016, 2017, 2018, 2019 and 2020, respectively. However, there was substantial regional variation in performance, with the average correlation in the East South Atlantic states being 0.81 while the average correlation in the Mountain states was only 0.45. The nowcast predictions displayed strong seasonal and geographic patterns, with predicted activity generally being highest in the summer months. CONCLUSIONS: The nowcast model is a valuable tool by which to educate pet owners regarding the risk of fleas and flea-borne disease and the need to routinely administer flea preventatives. While it is ideal for domestic cats and dogs to on flea preventatives year-round, many pets remain vulnerable to flea infestation. Alerting pet owners to the local increased risk of flea activity during certain times of the year may motivate them to administer appropriate routine preventives.

Ectoparasites of European hedgehogs (Erinaceus europaeus) in Germany and their health impact.

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.

Molecular detection and characterization of Bartonella spp. in small mammals in the Amazonia and Cerrado biomes, midwestern Brazil.

Although Bartonella spp. have been worldwide described in rodents and bats, few studies have reported these agents in marsupials. The present work aimed to investigate the occurrence and genetic diversity of Bartonella in small mammals (rodents, marsupials, and bats) and associated ectoparasites in two ecoregions (Amazonia and Cerrado biomes) in midwestern Brazil. For this purpose, DNA samples from 378 specimens of small mammals (128 rodents, 111 marsupials, and 139 bats) and 41 fleas (Siphonaptera) were screened for the Bartonella genus employing a quantitative real-time PCR assay (qPCR) based on the nuoG (nicotinamide adenine dinucleotide dehydrogenase gamma subunit) gene. Then, positive samples in qPCR were submitted to conventional PCR (cPCR) assays targeting the gltA, ftsZ, and rpoB genes. One (0.78 %) rodent, 23 (16.54 %) bats, and 3 (7.31 %) fleas showed positive results in the qPCR for Bartonella sp. After cPCR amplification and sequencing, 13 partial Bartonella DNA sequences of the following genes were obtained only from bats´ blood samples: 9 gltA (citrate synthase), 3 ftsZ (cell division protein), and 1 rpoB (RNA polymerase beta subunit). The maximum likelihood inference based on the gltA gene positioned the obtained sequences in three different clades, closely related to Bartonella genotypes previously detected in other bat species and bat flies sampled in Brazil and other countries from Latin America. Similarly, the ftsZ sequences clustered in two different clades with sequences described in bats from Brazil, other countries from Latin America, and Georgia (eastern Europe). Finally, the Bartonella rpoB from a specimen of Lophostoma silvicolum clustered with a Bartonella sp. sequence obtained from a Noctilio albiventris (KP715475) from French Guiana. The present study provided valuable insights into the diversity of Bartonella genotypes infecting bats from two ecoregions (Amazonia and Cerrado) in midwestern Brazil and emphasized that further studies should be conducted regarding the description and evaluation of different lineages of Bartonella in wild small mammals and their ectoparasites in different Brazilian biomes.

Surveillance of Fleas and Their Small Mammal Hosts for Plague Risks in Some Main Seaports of the Islands of the Southwestern Indian Ocean.

Since ancient times, seaports have been the hot spots for plague introduction into free countries. Infected ship rats reached new areas, and epizootics occurred prior to human infection via flea bites. Beginning in the 1920s/1930s, rodent and flea surveillance was carried out as part of plague hazard management in seaports of the world. Nowadays, such activity is not done regularly. In the southwestern Indian Ocean (SWIO) region, plague surveillance is of great importance given plague endemicity in Madagascar and thus the incurred risk for neighboring islands. This study reports animal-based surveillance aimed at identifying fleas and their small mammal hosts in SWIO seaports as well as Yersinia pestis detection. Small mammal trappings were performed in five main seaports of Madagascar (Toamasina and Mahajanga), Mauritius (Port Louis), and the Union of Comoros (Moroni and Mutsamudu). Mammals were euthanized and their fleas collected and morphologically identified before Y. pestis detection. In total, 145 mammals were trapped: the brown rat Rattus norvegicus (76.5%), the black rat Rattus rattus (8.3%), and the Asian house shrew Suncus murinus (15.2%). Fur brushing allowed collection of 1,596 fleas exclusively identified as Xenopsylla cheopis. All tested fleas were negative for Y. pestis DNA. This study shows that both well-known plague mammal hosts and flea vectors occur in SWIO seaports. It also highlights the necessity of carrying out regular animal-based surveillance for plague hazard management in this region.

Population dynamics of plague vector fleas in an endemic focus: implications for plague surveillance.

Plague is a zoonotic vector-borne disease caused by the bacterium Yersinia pestis. In Madagascar, it persists in identified foci, where it is a threat to public health generally from September to April. A more complete understanding of how the disease persists could guide control strategies. Fleas are the main vector for transmission between small mammal hosts and humans, and fleas likely play a role in the maintenance of plague. This study characterized the dynamics of flea populations in plague foci alongside the occurrence of human cases. From 2018 to 2020, small mammals were trapped at sites in the central Highlands of Madagascar. A total of 2,762 small mammals were captured and 5,295 fleas were collected. The analysis examines 2 plague vector species in Madagascar (Synopsyllus fonquerniei and Xenopsylla cheopis). Generalized linear models were used to relate flea abundance to abiotic factors, with adjustments for trap location and flea species. We observed significant effects of abiotic factors on the abundance, intensity, and infestation rate by the outdoor-associated flea species, S. fonquerniei, but weak seasonality for the indoor-associated flea species, X. cheopis. A difference in the timing of peak abundance was observed between the 2 flea species during and outside the plague season. While the present study did not identify a clear link between flea population dynamics and plague maintenance, as only one collected X. cheopis was infected, the results presented herein can be used by local health authorities to improve monitoring and control strategies of plague vector fleas in Madagascar.