Quantitative assessment of spicule length in Heligmosomoides spp. (Nematoda, Heligmosomidae): distinction between H. bakeri, H. polygyrus and H. glareoli.

Nematode spicules play a vital role in the reproductive activity of species that possess them. Our primary objective was to compare the lengths of spicules of the laboratory mouse (Mus musculus) ­ maintained isolate H. bakeri ­ with those of H. polygyrus from naturally infected wood mice (Apodemus sylvaticus). On a more limited scale, we also included H. glareoli from bank voles (Myodes glareolus), a species reputed to possess longer spicules than either of the 2 former species. In total, we measured 1264 spicules (H. bakeri, n = 614; H. polygyrus n = 582; and H. glareoli, n = 68). There was a highly significant difference between the spicule lengths of the Nottingham-maintained H. bakeri (mean = 0.518 mm) and H. polygyrus (0.598 mm) from 11 different localities across the British Isles. A comparison of the spicules of H. bakeri maintained in 4 different laboratories in 3 continents revealed a range in the mean values from 0.518 to 0.540 mm, while those of worms from Australian wild house mice were shorter (0.480 mm). Mean values for H. polygyrus from wood mice from the British Isles ranged from 0.564 to 0.635 mm, although isolates of this species from Norway had longer spicules (0.670 mm). In agreement with the literature, the spicules of H. glareoli were considerably longer (1.098 mm). Since spicules play a vital role in the reproduction of nematode species that possess them, the difference in spicule lengths between H. bakeri and H. polygyrus adds to the growing evidence that these 2 are quite distinct species and likely reproductively isolated.

Urban rodent reservoirs of Borrelia spp. in Warsaw, Poland.

The anticipated worldwide surge in urban environments is generating ever-greater interest in the study of host-pathogen interactions in this specific type of habitat. We investigated the potential of city-inhabiting rodents to serve as the main Lyme borreliosis agents (Borrelia spp.) reservoir. We also tried to verify if anthropogenic disturbances changing the vertebrate species community composition may also alter the scheme of Borrelia spp. circulation. A total of 252 Apodemus mice (A. agrarius, A. flavicollis, A. sylvaticus) were captured in Warsaw (Poland), at sites classified into different zones of anthropogenic disturbance, ranging from suburban forests to municipal parks strictly in the city centre. Borrelia spp. infection, ascertained based on bacterium DNA presence in the rodents' blood, was found only in A. agrarius and A. flavicollis (7.6 and 6%, respectively). Only one species from the Borrelia genus - the mammal-associated species B. afzelii - was found in the mice studied. We found no statistical evidence of a correlation between infection in Apodemus mice and the zone of anthropogenic disturbance where the mice were caught. Non-homogeneous concentrations of Borelia spp. infected specimens within the strict city centre area suggest a lack of contact between members of particular mice subpopulations, and their responsibility for relatively high, but local Borrelia spp. infection.

Molecular detection and phylogenetic analysis of Hepatozoon spp. in questing Ixodes ricinus ticks and rodents from Slovakia and Czech Republic.

By amplification and sequencing of 18S rRNA gene fragments, Hepatozoon spp. DNA was detected in 0.08 % (4/5057) and 0.04 % (1/2473) of questing Ixodes ricinus ticks from Slovakia and Czech Republic, respectively. Hepatozoon spp. DNA was also detected in spleen and/or lungs of 4.45 % (27/606) of rodents from Slovakia. Prevalence of infection was significantly higher in Myodes glareolus (11.45 %) than in Apodemus spp. (0.28 %) (P < 0.001). Sequencing of 18S rRNA Hepatozoon spp. gene amplicons from I. ricinus showed 100 % identity with Hepatozoon canis isolates from red foxes or dogs in Europe. Phylogenetic analysis showed that at least two H. canis 18S rRNA genotypes exist in Slovakia of which one was identified also in the Czech Republic. The finding of H. canis in questing I. ricinus suggests the geographical spread of the parasite and a potential role of other ticks as its vectors in areas where Rhipicephalus sanguineus is not endemic. Sequencing of 18S rRNA gene amplicons from M. glareolus revealed the presence of two closely related genetic variants, Hepatozoon sp. SK1 and Hepatozoon sp. SK2, showing 99-100 % identity with isolates from M. glareolus from other European countries. Phylogenetic analysis demonstrates that 18S rRNA variants SK1 and SK2 correspond to previously described genotypes UR1 and UR2 of H. erhardovae, respectively. The isolate from Apodemus flavicollis (Hepatozoon sp. SK3b) was 99 % identical with isolates from reptiles in Africa and Asia. Further studies are necessary to identify the taxonomic status of Hepatozoon spp. parasitizing rodents in Europe and the host-parasite interactions in natural foci.

Neoehrlichiosis: an emerging tick-borne zoonosis caused by Candidatus Neoehrlichia mikurensis.

Candidatus Neoehrlichia mikurensis is an emerging tick-borne pathogen causing a systemic inflammatory syndrome mostly in persons with underlying hematologic or autoimmune diseases. As it is neither well-known nor well-recognized, it might be misdiagnosed as recurrence of the underlying disease or as an unrelated arteriosclerotic vascular event. The pathogen is transmitted by hard ticks of the genus Ixodes and is closely associated with rodents in which transplacental transmission occurs. Transovarial transmission in ticks has not yet been shown. Infection rates vary greatly in ticks and rodents, but the causes for its spatiotemporal variations are largely unknown. This review summarizes the current state of knowledge on the geographical distribution and clinical importance of Ca. N. mikurensis. By elucidating the life history traits of this pathogen and determining more accurately its incidence in the human population, a better assessment of its public health relevance can be made. Most urgent research needs are the in vitro-cultivation of the pathogen, the development of specific serological tests, the determination of the full genomic sequence, the routine implementation of molecular diagnosis in diseased patients with a particular panel of underlying diseases, and promoting the knowledge about neoehrlichiosis among general practitioners, hospital physicians and the risk groups such as forest workers or immune-compromised people to raise awareness about this disease that can easily be treated when correctly diagnosed.

Response of wild rodents to red fox feces: implication for the echinococcus infection.

Echinococcus multilocularis causes zoonotic disease, alveolar echinococcosis. The life cycle of E. multilocularis is maintained by the predator-prey relationship between red foxes and rodents. Infection to red fox (Vulpes vulpes) of E. multilocularis is considered that rodents take eggs of E. multilocularis, then red fox forage the rodents. However, it has been not known how to take eggs by rodents. On infection process of E. multilocularis from red foxes to rodents, we predicted that rodents would forage or touch with feces of red fox to use undigested materials within the feces. We monitored rodent's response to fox feces and their distance to the feces by using camera trap from May to October 2020. Myodes spp. and Apodemus spp. touched fox feces, and touch rate of Apodemus spp. was significantly higher than that of Myodes spp. We found smelling and passing as contact behaviors to fox feces by Myodes spp., while Apodemus spp. showed behaviors which oral directly contacted feces. There was no significant difference on the shortest distance between Apodemus spp. and Myodes spp. The distance between 0 cm and 5 cm was mostly observed for both rodents. The results that Myodes spp. did not forage feces and their contact to feces was low frequency suggested that the infection from red foxes to Myodes spp., the main intermediate host, was to be other pathways. The approach to feces and the act near feces might increase the probability attached with eggs.

High Frequency of Apodemus Mice Boosts Inverse Activity Pattern of Bank Voles, Clethrionomys glareolus, through Non-Aggressive Intraguild Competition.

Sympatric animals with similar requirements can separate their ecological niches along the microhabitat, food and time axes. There may be alternative reasons for an interspecific different activity pattern, such as intraspecific social constraints, predator avoidance or physical conditions such as temperature, precipitation and illumination. We investigated the importance of intraguild competition in a 2-year study in an inner-alpine mixed forest, using small forest rodents as our model species. Apodemus mice were the physically superior, and bank voles, Clethrionomys glareolus, the inferior competitor. We predicted that bank voles would exhibit increased diurnal activity when frequencies of the almost exclusively nocturnal Apodemus mice were high during a seed mast year. To investigate this, we recorded 19,138 1 min videos. Controlling for confounding variables, bank vole diurnal activity was significantly related to the frequency of Apodemus mice. We assume that at high densities of Apodemus mice, a purely nocturnal separation in the niche dimensions of time, habitat and microhabitat is no longer sufficient, and therefore an inverse activity pattern by the bank voles is reinforced. Our videos showed, however, that this does not require persistent aggressive meetings and we explain this by the long co-evolution of the taxa under study.

First Record of Hepatozoon spp. in Alpine Wild Rodents: Implications and Perspectives for Transmission Dynamics across the Food Web.

Among the Apicomplexa parasites, Hepatozoon spp. have been mainly studied in domestic animals and peri-urban areas. The epidemiology of Hepatozoon spp. is poorly investigated in natural systems and wild hosts because of their scarce veterinary and economic relevance. For most habitats, the occurrence of these parasites is unknown, despite their high ecosystemic role. To fill this gap for alpine small mammals, we applied molecular PCR-based methods and sequencing to determine the Hepatozoon spp. in 830 ear samples from 11 small mammal species (i.e., Apodemus, Myodes, Chionomys, Microtus, Crocidura and Sorex genera) live-trapped during a cross-sectional study along an altitudinal gradient in the North-Eastern Italian Alps. We detected Hepatozoon spp. with an overall prevalence of 35.9%. Two species ranging from 500 m a.s.l. to 2500 m a.s.l. were the most infected: My. glareolus, followed by Apodemus spp. Additionally, we detected the parasite for the first time in another alpine species: C. nivalis at 2000-2500 m a.s.l. Our findings suggest that several rodent species maintain Hepatozoon spp. along the alpine altitudinal gradient of habitats. The transmission pathway of this group of parasites and their role within the alpine mammal community need further investigation, especially in consideration of the rapidly occurring environmental and climatic changes.

Borrelia miyamotoi infection in Apodemus spp. mice populating an urban habitat (Warsaw, Poland).

The two of three species of city-inhabiting (Warsaw, Poland) Apodemus spp. mice were showed to be infected with Borrelia miyamotoi, spirochete bacterium considered to be a tick-borne relapsing fever (TBRF) disease causative agent. The pathogen infection was ascertained based on bacterium DNA presence in a blood (obtained from the lateral tail vein) of the mice, using the nested polymerase chain reaction (PCR) technique. The B. miyamotoi carriers populated not only green spaces situated in the suburban areas but also recreational parks and lawns in the city centre proper. Moreover, we found no significant differences in the infection rate depending on mouse species. The total prevalence of infection at the level of 5.6% suggests the potential role of urban-settled rodents in the spreading of the tick-borne zoonosis, which pose high risk to public health.

Differential spatial responses of rodents to masting on forest sites with differing disturbance history.

Mast seeding, the synchronized interannual variation in seed production of trees, is a well-known bottom-up driver for population densities of granivorous forest rodents. Such demographic effects also affect habitat preferences of the animals: After large seed production events, reduced habitat selectivity can lead to spillover from forest patches into adjacent alpine meadows or clear-cuts, as has been reported for human-impacted forests. In unmanaged, primeval forests, however, gaps created by natural disturbances are typical elements, yet it is unclear whether the same spillover dynamics occur under natural conditions. To determine whether annual variation in seed production drives spillover effects in naturally formed gaps, we used 14 years of small mammal trapping data combined with seed trap data to estimate population densities of Apodemus spp. mice and bank voles (Myodes glareolus) on 5 forest sites with differing disturbance history. The study sites, located in a forest dominated by European beech (Fagus sylvatica), Norway spruce (Picea abies), and silver fir (Abies alba), consisted of two primeval forest sites with small canopy gaps, two sites with larger gaps (after an avalanche event and a windthrow event), and a managed forest stand with closed canopy as a control. Hierarchical Bayesian N-mixture models revealed a strong influence of seed rain on small rodent abundance, which were site-specific for M. glareolus but not for Apodemus spp. Following years of moderate or low seed crop, M. glareolus avoided open habitat patches but colonized those habitats in large numbers after full mast events, suggesting that spillover events also occur in unmanaged forests, but not in all small rodents. The species- and site-specific characteristics of local density responding to food availability have potentially long-lasting effects on forest gap regeneration dynamics and should be addressed in future studies.

Host-parasite interactions of rodent hosts and ectoparasite communities from different habitats in Germany.

BACKGROUND: Small mammals are important maintenance hosts of ectoparasites as well as reservoir hosts for many arthropod-borne pathogens. In Germany, only a few studies have investigated ectoparasite communities on small mammals in their natural habitats. The aim of this study was to assess the species diversity and parameters influencing the mean intensity and prevalence of macroscopically visible ectoparasites, such as fleas, predatory mites and ticks. METHODS: A total of 779 small mammals and 3383 ticks were available from earlier investigations for the data analysis of the current study from three differently structured study sites. In addition, fleas and predatory mites were collected from the captured rodents and taxonomically identified. Regression analyses were conducted on the group (ticks/mites/fleas) and species levels using hurdle models for the abundance of ectoparasite groups and a negative binomial model for the abundance of species. RESULTS: Nearly 90% of the small mammals analyzed were infested with ectoparasites, with an average of 7.3 specimens per host. Hosts were infested with up to six species of ectoparasites simultaneously. In total, 12 flea, 11 mite and three tick species were detected. Ticks were more prevalent than fleas or mites, with > 80% of the hosts in urban and forest areas hosting ticks and around 60% of hosts presenting fleas, and only 20-40% of hosts presenting mites. Polyparasitism had a statistically significant influence on the prevalence of the investigated tick, mite and flea species, with odds ratios of > 1.0. Trapping location, season and host characteristics had significant influences on some-but not all-of the investigated species. CONCLUSIONS: The diversity of flea species was unexpectedly high and higher than that reported in comparable studies, which can be explained by the differently structured habitats and regions examined in this study. Polyparasitism was a key influencing factor and had a positive effect on the prevalence and/or abundance of the predominant tick, flea and mite species occurring on small mammals. Season, trapping location, host species and sex of the host species also had an influence on the prevalence and mean intensity of certain, but not all, ectoparasite species.

Range expansion of Ixodes ricinus to higher altitude, and co-infestation of small rodents with Dermacentor marginatus in the Northern Apennines, Italy.

Immature ticks (Ixodes ricinus and Dermacentor marginatus) were collected from small rodents (Apodemus spp. and Myodes glareolus), in the Northern Apennines, Italy, at an altitude up to 1650 m above sea level (a.s.l.), from 2009 through 2012. While D. marginatus had been found at the same location in studies carried out in 1994, I. ricinus was very rare or absent. Prevalence (95% confidence interval) of infestation by I. ricinus larvae on Apodemus spp. was 54.4% (47.5, 61.2), and it was greater than prevalence of D. marginatus larvae on the same hosts (23.3%, 17.8, 29.5). The mean (standard deviation) numbers of I. ricinus and D. marginatus larvae per individual Apodemus spp. were similar: 2.3 (4.1) and 2.1 (9.8), respectively. The monthly infestation pattern of the two tick species on Apodemus spp. were different. I. ricinus larvae were more frequent in June and September, than in July-August. I. ricinus nymphs were generally rare, and were most frequently found in July. The prevalence of D. marginatus larvae peaked in July-August, whereas nymphs were mostly active in August-September. Increasing population densities of roe deer (Capreolus capreolus), and increasing temperatures, in the last decades, in the Apennine area might have contributed to the observed range expansion of I. ricinus.

Rickettsia slovaca in immature Dermacentor marginatus and tissues from Apodemus spp. in the northern Apennines, Italy.

Immature Dermacentor marginatus ticks and tissues from small rodents were tested for infection with Rickettsia slovaca in the northern Apennines, Lucca Province, where tick-borne lymphadenopathy (TIBOLA) was previously reported in people. Prevalence of infestation with D. marginatus was 30.5% (n=131, 95% CI: 22.8-39.2%) in Apodemus spp. and 26.5% (n=34, 95% CI: 12.9-44.4%) in Myodes glareolus, which were captured during 1980 trap nights in 2009 and 2010. Rickettsia slovaca was identified by polymerase chain reaction, targeting the gltA and OmpA genes, in ear biopsies from 8 out of 37 tested Apodemus (22%, 95% CI: 9.8-38.2%), but not from 9 M. glareolus. The prevalence of R. slovaca in D. marginatus feeding on Apodemus spp. was 53% in larvae (n=51, 95% CI: 38.5-67.1%) and 47.5% in nymphs (n=59, 95% CI: 34.3-60.9%). No larvae (0.0%, 95% CI: 0-36.9%), but one nymph removed from M. glareolus was positive (10%, 95% CI: 0.3-44.5%). Prevalence of R. slovaca in host-seeking D. marginatus larvae, collected in the same area, was 42% (n=38; 95% CI: 26.3-59.2%). Prevalence of R. slovaca was greater in larvae feeding on PCR-positive Apodemus than in those feeding on negative mice (78.6% vs. 37.1%). Furthermore, levels of infestation with D. marginatus larvae were greater for R. slovaca-positive mice. The infection of Apodemus spp. was probably the result of repeated bites by transovarially infected larvae. On the other hand, the finding of R. slovaca in mice tissues would be compatible with transmission from these hosts to feeding D. marginatus. Based on such a hypothesis, the most heavily infested Apodemus might play a role as amplifiers of the infection.