Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in small rodents and attached ticks in the Northern Apennines, Italy.

Ticks and ear biopsies were collected from wild small rodents in 2011 and 2012 in the northern Apennines (Italy), up to 1650 m above sea level. Apodemus spp. (n = 83) and Myodes glareolus (n = 22) were infested by Ixodes ricinus (192 larvae and two nymphs), Dermacentor marginatus (179 larvae and 29 nymphs), and Ixodes trianguliceps (three larvae and two nymphs). We detected several Borrelia burgdorferi sensu lato (s.l.) genospecies (B. afzelii, B. burgdorferi sensu stricto, B. garinii, B. lusitaniae, B. valaisiana) in I. ricinus and skin biopsies. The most common genospecies found in I. ricinus was B. valaisiana, while it was B. lusitaniae in tissues. Spotted fever group (SFG) rickettsiae (Rickettsia monacensis, R. slovaca and R. raoultii) infected I. ricinus, D. marginatus and rodent tissues. Rickettsia slovaca was the Rickettsia species most frequently found in our samples. Coinfections by B. burgdorferi s.l. and SFG rickettsiae indicate an overlap of transmission cycles and potential risk for humans to be infected by multiple pathogens, resulting in more severe symptoms. The findings of B. lusitaniae and R. slovaca in bank voles, and of B. valaisiana in small rodents, open new questions about host-pathogen interactions. In addition, our results highlight the importance of small rodents as data sources for studying tick-borne pathogens.

Analysis of the environmental and host-related factors affecting the distribution of the tick Dermacentor marginatus.

Understanding and responding to the ecological, social and economic conditions that facilitate disease emergence and transmission represents a substantial challenge for epidemiologists and health professionals. In this article we integrate knowledge about the human and the vector population, to provide a context from which to examine the underlying causal factors of D. marginatus-borne diseases emergence in the study area. Within this framework we analyse the biotic and abiotic factors that drive D. marginatus population dynamics and the role of its typical host for dispersal. These investigations suggest that D. marginatus is a tick species prone to spatially overlap its presence with human population presence. Then we consider the public health implications for the residents, when simply carrying out trivial outdoor activities may increase the risk to contact an infected tick.

Presence of host-seeking Ixodes ricinus and their infection with Borrelia burgdorferi sensu lato in the Northern Apennines, Italy.

Host-seeking ticks were collected in the Northern Apennines, Italy, by dragging at 35 sites, at altitudes ranging from 680 and 1670 m above sea level (asl), from April to November, in 2010 and 2011. Ixodes ricinus (4431 larvae, 597 nymphs and 12 adults) and Haemaphysalis punctata (11,209 larvae, 313 nymphs, and 25 adults) were the most abundant species, followed by Haemaphysalis sulcata (20 larvae, five nymphs, and 13 adults), Dermacentor marginatus (42 larvae and two adults) and Ixodes hexagonus (one nymph). Greatest numbers of ticks were collected at locations characterised by southern exposure and limestone substratum, at altitudes <1400 m asl; I. ricinus was most abundant in Turkey oak (Quercus cerris) wood, whereas H. punctata was mostly collected in hop hornbeam (Ostrya carpinifolia) wood and on exposed rocks. Ixodes ricinus was also found up to 1670 m asl, in high stand beech (Fagus sylvatica) wood. The overall prevalence of Borrelia burgdorferi sensu lato (sl) in 294 host-seeking I. ricinus nymphs was 8.5 %. Borrelia garinii was the most frequently identified genospecies (64.0 % of positive nymphs), followed by B. valaisiana, B. burgdorferi sensu stricto, B. afzelii, and B. lusitaniae. Based upon the comparison with the results of previous studies at the same location, these research findings suggest the recent invasion of the study area by the tick vector and the agents of Lyme borreliosis.

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.

Transmission dynamics of Borrelia lusitaniae and Borrelia afzelii among Ixodes ricinus, lizards, and mice in Tuscany, central Italy.

To estimate the basic reproduction number (R(0)) of Borrelia lusitaniae and Borrelia afzelii, we formulated a mathematical model considering the interactions among the tick vector, vertebrate hosts, and pathogens in a 500-ha enclosed natural reserve on Le Cerbaie hills, Tuscany, central Italy. In the study area, Ixodes ricinus were abundant and were found infected by B. lusitaniae and B. afzelii. Lizards (Podarcis spp.) and mice (Apodemus spp.), respectively, are the reservoir hosts of these two Borrelia burgdorferi sensu lato (s.l.) genospecies and compete for immature ticks. B. lusitaniae R(0) estimation is in agreement with field observations, indicating the maintenance and diffusion of this genospecies in the study area, where lizards are abundant and highly infested by I. ricinus immature stages. In fact, B. lusitaniae shows a focal distribution in areas where the tick vector and the vertebrate reservoir coexist. Mouse population dynamics and their relatively low suitability as hosts for nymphs seem to determine, on the other hand, a less efficient transmission of B. afzelii, whose R(0) differs between scenarios in the study area. Considering host population dynamics, the proposed model suggests that, given a certain combination of the two host population sizes, both spirochete genospecies can coexist in our study area. Additional incompetent hosts for B. burgdorferi s.l. have a negative effect on B. afzelii maintenance, whose R(0) results > 1 only with high mouse population densities and/or low lizards abundance, but they do not seem to influence B. lusitaniae transmission cycle on Le Cerbaie. Secondly, our model confirms the importance of nymphs' infestation, of host population density and diversity, and spirochetes host association for the maintenance of the transmission cycle of B. burgdorferi s.l.

Modeling the spread of vector-borne diseases on bipartite networks.

BACKGROUND: Vector-borne diseases for which transmission occurs exclusively between vectors and hosts can be modeled as spreading on a bipartite network. METHODOLOGY/PRINCIPAL FINDINGS: In such models the spreading of the disease strongly depends on the degree distribution of the two classes of nodes. It is sufficient for one of the classes to have a scale-free degree distribution with a slow enough decay for the network to have asymptotically vanishing epidemic threshold. Data on the distribution of Ixodes ricinus ticks on mice and lizards from two independent studies are well described by a scale-free distribution compatible with an asymptotically vanishing epidemic threshold. The commonly used negative binomial, instead, cannot describe the right tail of the empirical distribution. CONCLUSIONS/SIGNIFICANCE: The extreme aggregation of vectors on hosts, described by the power-law decay of the degree distribution, makes the epidemic threshold decrease with the size of the network and vanish asymptotically.

Temporal variations in the usefulness of normalized difference vegetation index as a predictor for Ixodes ricinus (Acari: Ixodidae) in a Borrelia lusitaniae focus in Tuscany, central Italy.

Host-seeking ticks were collected during monthly dragging sessions from November 2004 through October 2006 in Tuscany, central Italy. Normalized difference vegetation index (NDVI), which was calculated from Landsat ETM+ 7 remote sensing data recorded in August 2001, was significantly correlated with numbers of host-seeking immature Ixodes ricinus L. (Acari: Ixodidae) during periods of relatively low rainfall such as summer 2005 (Spearman's p = 0.78, P < 0.001 for nymphs in July) and to a lower extent in spring-summer 2006. In spring 2005, when rainfall was relatively high, the correlation was weak and not statistically significant. Generalized estimating equations (GEEs), taking into account repeated sampling of the same dragging sites, were used to model the effects of NDVI and season on counts of host-seeking I. ricinus nymphs. Seasonal variations of the effect of NDVI yielded a significant NDVI-by-season interaction in the first year of the study (November 2004-October 2005), but not in the second year (November 2005-October 2006) when there was a 2.5-fold increase of the number of nymphs per 100-m dragging for every 0.1 unit increase in NDVI (95% confidence interval = 1.6, 3.0). Risk maps that were obtained based on GEE results confirmed that the predicted number of I. ricinus nymphs per 100 m was relatively homogeneous through the study area during the 2005 spring peak of activity. Conversely, in 2006, the predicted abundance of nymphs was greater in moist bottomland habitat (characterized by high NDVI) than in dry, typically Mediterranean, upland habitat.

Borrelia lusitaniae in immature Ixodes ricinus (Acari: Ixodidae) feeding on common wall lizards in Tuscany, central Italy.

Lizards and small rodents were live captured in Tuscany, central Italy, from May through August 2005. Prevalence of infestation by larval Ixodes ricinus L. (Acari: Ixodidae) and mean numbers of larvae per host were not significantly different for common wall lizards, Podarcis muralis Laurenti, and Apodemus spp. mice, whereas infestation levels by nymphs were significantly greater on lizards. Borrelia lusitaniae, which was previously shown to be dominant in host-seeking I. ricinus in the same study area, was detected by polymerase chain reaction (PCR) in 19.8% (95% confidence interval: 14.4, 26.0) of larval ticks and in 52.9% (27.8, 77.0) of nymphs that were collected from lizards. Moreover, 18.8% (7.2, 36.4) and 25.0 (3.2, 65.1) of lizards' tail biopsies and blood samples, respectively, were positive for B. lusitaniae. Conversely, attached ticks and ear biopsies from Apodemus spp. mice were PCR negative. Passerine birds belonging to 10 species were live captured in March 2005, and Borrelia valaisiana was detected in 57.1% (18.4, 90.1) of I. ricinus nymphs feeding on Eurasian blackbirds, Turdus merula L. Results of this study suggest that lizards play an important role as reservoirs for B. lusitanae and may affect the dominance of this genospecies in the Mediterranean area.