Detection of Rickettsia hulinii in ticks of the Haemaphysalis concinna species in Russia

The presence of Rickettsia hulinii was detected in ticks of the Haemaphysalis concinna species in the Asiatic part of Russia. It was the first detection of these rickettsiae outside the territory of the Chinese People's Republic. This tick is spread over a broken area and may occur in different regions of Eurasia. At present at least 3 rickettsial species of the tick-borne spotted fever group were found in Ixodes ticks on the territory of Eastern Siberia: R. sibirica, known to be the etiological agent of tick-borne rickettsiosis, R. hulinii with proved pathogenicity for laboratory animals and R.DnS14 with no data available on its pathogenicity.     

Rickettsia slovaca and Rickettsia raoultii in Dermacentor marginatus and Dermacentor reticulatus ticks from Slovak Republic

Rickettsiae, obligate intracellular Gram-negative bacteria, responsible for mild to severe diseases in humans are associated with arthropod vectors. Dermacentor marginatus and Dermacentor reticulatus are known vectors of Rickettsia slovaca and Rickettsia raoultii distributed across Europe. A total of 794 D. marginatus, D. reticulatus and Ixodes ricinus adult ticks were collected from the vegetation, removed from horses, sheep, goats and dogs in Slovakia. The DNA of Rickettsia sp. was found in 229 ticks by PCR amplifying parts of gltA, ompA and sca4 genes. Next analyses of Rickettsia-positive samples by PCR-RFLP and/or sequencing showed D. reticulatus ticks were more infected with R. raoultii and D. marginatus were more infected with R. slovaca. The prevalence of R. raoultii was 8.1-8.6% and 22.3-27% in D. marginatus and D. reticulatus, respectively. The prevalence of R. slovaca was 20.6-24.3% in D. marginatus and 1.7-3.4% in D. reticulatus. Intracellular growth of R. raoultii isolate from D. marginatus tick was evaluated by rOmpA-based quantitative SybrGreen PCR assay. The highest point of multiplication was recorded on the 7th and 8th day postinfection in Vero and L929 cells, respectively. R. raoultii was transmitted during feeding of R. raoultii-positive ticks to guinea pigs and subsequently rickettsial infection was recorded in all organs, the highest infection was in spleen, liver and heart. Our study describes the detection and isolation of tick-borne pathogens R. raoultii and R. slovaca, show that they are spread in Slovakia and highlight their risk for humans.     

Molecular detection of Babesia canis in Dermacentor reticulatus ticks collected in Slovakia

Dermacentor reticulatus ticks are recognized as the most important vectors of Babesia canis, the aetiological agent of canine babesiosis occurring throughout Europe. Vector competence of D. reticulatus for B. canis is well described and experimentally determined; however, by using molecular analysis it was proven so by one recent study in Russia. Herein, the additional molecular evidence of B. canis infection in D. reticulatus ticks collected in Slovakia is provided. Using PCR followed by sequencing of distinctive amplicons we determined the presence of Babesia canis canis in one of 100 tested adult ticks. Two zoonotic pathogens, Francisella tularensis and Coxiella burnetii, were previously isolated from D. reticulatus ticks in Slovakia. In our samples, we detected only the presence of F. tularensis.     

Genomic comparison of Rickettsia helvetica and other Rickettsia species

We report the complete and annotated genome sequence of Rickettsia helvetica strain C9P9, which was first isolated in 1979 from Ixodes ricinus ticks in Switzerland and is considered a human pathogen.     

Molecular identification of Rickettsia felis in ticks and fleas from an endemic area for Brazilian Spotted Fever

Rickettsioses are arthropod-borne diseases caused by parasites from the Order Rickettsiales. The most prevalent rickettsial disease in Brazil is Brazilian Spotted Fever (BSF). This work intends the molecular detection of those agents in ectoparasites from an endemic area of BSF in the state of Espírito Santo. A total of 502 ectoparasites, among them Amblyomma cajennense, Amblyomma dubitatum (A. cooperi), Riphicephalus sanguineus, Anocentor nitens and Ctenocephalides felis, was collected from domestic animals and the environment and separated in 152 lots according to the origin. Rickettsia sp. was detected in pools of all collected species by amplification of 17 kDa protein-encoding gene fragments. The products of PCR amplification of three samples were sequenced, and Rickettsia felis was identified in R. sanguineus and C. felis. These results confirm the presence of Rickettsia felis in areas previously known as endemic for BSF, disease caused by Rickettsia rickettsii. Moreover, they show the needing of further studies for deeper knowledge of R. felis-spotted fever epidemiology and differentiation of these diseases in Brazil.     

Spotted fever group Rickettsia in ticks from southeastern Spain natural parks

During an 8-years study, we collected from vegetation or domestic and wild mammals 1246 ticks (624 males, 511 females and 111 nymphs) belonging to 13 species in Jaen province (Andalusia) and we analyzed these ticks by PCR and sequencing for the presence of rickettsiae. Specific rickettsiae DNA was detected in 243 (19.5%) of the ticks tested. Sequence analysis of amplicons of gltA, ompA and ompB genes revealed that Ixodes ricinus were infected with R. monacensis, including strain IRS3, and R. helvetica (prevalences of 27.0% and 2.7%, respectively), while in I. ventalloi we found only this last species (12.5%). Moreover, Dermacentor marginatus presents R. slovaca (24.7%) and R. raoultii (59.9%). In Rhipicephalus sanguineus group ticks (Rh. sanguineus, Rh. turanicus and Rh. pusillus) only R. massiliae (15.2%) was found. Haemaphysalis punctata and Ha. sulcata were infected with a Rickettsia sp. near R. hoogstraalii (prevalence of 3.1% and 16.1%, respectively). In addition, Ha. punctata appeared infected with R. monacensis-like Rickettsia (1.0%) and R. raoultii (9.3%). None of I. hexagonus, Hyalomma lusitanicum, Hyalomma sp., Ha. hispanica or Rh. bursa studied ticks contained rickettsiae.     

Use of DNA sequences for Rickettsia identification in Ixodes ricinus ticks: the first detection of Rickettsia monacensis in Poland

The cosmopolitan tick Ixodes ricinus inhabiting Europe, including Poland, is a vector for many pathogens, such as various Rickettsia species, which spread to new territories. They are present mainly in the Mediterranean countries, but have also been found in Central Europe at increasing frequency. In the present study, the gltA gene, encoding citrate synthase, and an internal transcribed spacer (ITS) were employed to detect the DNA and identify the species of tick-borne pathogens of the Rickettsia genus. The presence of bacterial DNA was detected in 9.5% of the examined I. ricinus individuals. Based on the nucleotide sequences of the analysed genomic fragments, most pathogens were identified as Rickettsia helvetica, while Rickettsia monacensis was revealed in one case. We have described for the first time, to our knowledge, the occurrence of this species in Poland. Both markers employed in the experiments were successful in species identification of R. helvetica. The newly described species R. monacensis may be identified by the protein-coding gene, but the ITS nucleotide sequences proved insufficient.     

Isolation and identification of Rickettsia massiliae from Rhipicephalus sanguineus ticks collected in Arizona

Twenty Rhipicephalus sanguineus ticks collected in eastern Arizona were tested by PCR assay to establish their infection rate with spotted fever group rickettsiae. With a nested PCR assay which detects a fragment of the Rickettsia genus-specific 17-kDa antigen gene (htrA), five ticks (25%) were found to contain rickettsial DNA. One rickettsial isolate was obtained from these ticks by inoculating a suspension of a triturated tick into monolayers of Vero E6 monkey kidney cells and XTC-2 clawed toad cells, and its cell culture and genotypic characteristics were determined. Fragments of the 16S rRNA, GltA, rOmpA, rOmpB, and Sca4 genes had 100%, 100%, 99%, 99%, and 99%, respectively, nucleotide similarity to Rickettsia massiliae strain Bar29, previously isolated from R. sanguineus in Catalonia, Spain (L. Beati et al., J. Clin. Microbiol. 34:2688-2694, 1996). The new isolate, AZT80, does not elicit cytotoxic effects in Vero cells and causes a persistent infection in XTC-2 cells. The AZT80 strain is susceptible to doxycycline but resistant to rifampin and erythromycin. Whether R. massiliae AZT80 is pathogenic or infectious for dogs and humans or can cause seroconversion to spotted fever group antigens in the United States is unknown.     

Patterns of species richness in dry grassland patches in an agricultural landscape

Eighty-five patches of semi-natural grassland of varying size scattered in a agricultural landscape were investigated for their flora of vascular plants. Relationships between species richness and patch area, spatial isolation and local habitat conditions including heterogeneity were examined. Differences between single species and among groups of species defined by life-history traits were also investigated.
Area was shown to be an important determinant of species richness irrespective of habitat heterogeneity. Isolation in space and habitat heterogeneity also play significant roles. These results are consistent with results from a multitude of studies on fragments of ancient deciduous woodland in northern Europe, They are, however, contradictory to results from previous studies in grasslands within the same region. Seed mass and dispersal syndrome were poor predictors of the degree to which the species were affected by isolation of grassland patches. Seed mass deviation from community median could explain a small percentage of the variation in regional abundance. Logistic regression on species occurrences showed that few species are associated with large patches, and less than half seem to avoid isolated patches.     

Spatial dynamics of plant species in an agricultural landscape in the Netherlands

This study examined the changes in distribution patterns of 13 herbaceous plant species from 1998 to 2000 in ditch banks along the edges of arable fields in the Netherlands. The objective was to test if spatial dynamics could be related to spatial isolation and disturbance of habitat and to the dispersal and seed bank characteristics of the species. Knowledge of these relations should be used to increase the effectivity of agri-environmental schemes aiming at an increase of botanical diversity. All species frequently colonized empty patches and populations in occupied patches frequently went extinct. Most colonization events occurred within 50 m of conspecific source patches in the preceding year, but colonization events in patches at distances more than 200 m from conspecific source patches were also observed. The colonization probabilities decreased with isolation distance. For nine species this relation was statistically significant, after correction for year and habitat. The extinction probabilities increased with isolation. For only four species this relation was statistically significant. Both colonization and extinction probabilities were more often statistically significant related to isolation for species with transient seed banks than species with persistent seed banks. Implications for management options aiming at survival of plant species in fragmented landscapes are discussed.     

Seasonal and storm event nutrient removal by a created wetland in an agricultural watershed

This study examines the effectiveness of a 1.2-ha created/restored emergent marsh at reducing nutrients from a 17.0 ha agricultural and forested watershed in the Ohio River Basin in west central Ohio, USA, during base flow and storm flow conditions. The primary source of water to the wetland was surface inflow, estimated in water year 2000 (October 1999–September 2000) to be 646 cm/year. The wetland also received a significant amount of groundwater discharge at multiple locations within the site that was almost the same in quantity as the surface flow. The surface inflow had 2-year averages concentrations of 0.79, 0.033, and 0.16 mg L⁻¹ for nitrate + nitrite (as N), soluble reactive phosphorus (SRP), and total phosphorus (TP), respectively. Concentrations of nitrate–nitrite, SRP, and TP were 40, 56, and 59% lower, respectively, at the outflow than at the inflow to the wetland over the 2 years of the study. Concentrations of SRP and TP exported from the wetland increased significantly (α = 0.05) during precipitation events in 2000 compared to dry weather flows, but concentrations of nitrate–nitrite did not increase significantly. During these precipitation events the wetland retained 41% of the nitrate–nitrite, 74% of the SRP, and 28% of the TP (by mass). The wetland received an average of 50 g N m⁻² per year of nitrate–nitrite and 7.1 g m⁻² per year of TP in 2000. Retention rates for the wetland were 39 g N m⁻² per year of nitrates and 6.2 g P m⁻² per year. These are close to rates suggested in the literature for sustainable non-point source retention by wetlands. The design of this wetland appears to be suitable as it retained a significant portion of the influent nutrient load and did not lose much of its retention capacity during heavy precipitation events. Some suggestions are given for further design improvements.     

Ecology and phenology of ticks in Zambia: Seasonal dynamics on cattle

A study of the seasonality and infestation rates of ticks was carried out in 11 cattle herds in different ecological habitats in Zambia between 1980 and 1982. Wherever possible supplementary data were obtained from opportunistic collections from cattle and other hosts.Analysis of over 1000 tick collections from cattle indicated that infestation rates of the most important species,Amblyomma variegatum andRhipicephalus appendiculatus vary in different ecological habitats: (i) In Western Province, infestations are much lower than elsewhere; (ii) in Central and Southern Provinces, moderate to high infestations occur; and (iii) in Eastern Province,R. appendiculatus numbers are generally low andA. variegatum numbers are moderate.These two species, however, have similar life cycles throughout their range with one generation per year. Larvae occur mainly from March to May, nymphae from May to September, and adults ofA. variegatum from October to December and ofR. appendiculatus from December to April. Boophilus decoloratus appears to have two to four generations per year but is uncommon during the rainy season. In some areas in central ZambiaRhipicephalus compositus adults are seasonally common in September–October whereasRhipicephalus evertsi is more or less ubiquitous. Low to moderate infestations ofHyalomma truncatum andHyalomma rufipes occur in most areas.At least 14 other less common or rare species ofRhipicephalus, Amblyomma, Haemaphysalis andIxodes were taken infrequently from cattle. These and other host-specific species were also collected from dogs, sheep, various wildlife hosts and the environment.Infestation rates, seasonality and host-relationship of tick species are discussed in relation to their ecology. Relevant biosystematic and disease relationships are reviewed briefly. The baseline data derived from this study are adequate for integrated analysis with those from other ecological and economic investigations to formulate tick control strategies.     

Seasonal comparisons of Barn Owl diets in an agricultural and natural area in central South Africa

Longevity and displacement records aid in the understanding of animal senescence and the possible range of a species. Within wild populations this information is infrequent with most longevity reports being based on captive individuals and displacements being calculated from few monitored individuals within the population. Ringing offers a means of calculating these values and for avian species SAFRING and other bird ringing schemes are critically important. As many vulture populations are under threat, it has become ever more important to analyse all aspects of their demography to understand what can be done to aid the conservation of these species. Southern Africa’s eight vulture species are almost all listed as globally threatened and as a result many studies incorporate this ringing information. This has allowed for longevity records to be calculated for most of the species with the Cape Vulture obtaining the greatest reported longevity of 31 years 2 months and 14 days. In addition to the longevity records, ringing also produces maximum displacement records with the greatest vulture displacement being 2 530 km (White-backed Vulture). Although vulture ringing effort has improved in recent years very few new longevity records appear to be produced with most recent recoveries being under 10 years.     

Rickettsia monacensis sp. nov., a spotted fever group Rickettsia,from ticks (Ixodes ricinus) collected in a European city park

We describe the isolation and characterization of Rickettsia monacensis sp. nov. (type strain, IrR/Munich(T)) from an Ixodes ricinus tick collected in a city park, the English Garden in Munich, Germany. Rickettsiae were propagated in vitro with Ixodes scapularis cell line ISE6. BLAST analysis of the 16S rRNA, the citrate synthase, and the partial 190-kDa rickettsial outer membrane protein A (rOmpA) gene sequences demonstrated that the isolate was a spotted fever group (SFG) rickettsia closely related to several yet-to-be-cultivated rickettsiae associated with I. ricinus. Phylogenetic analysis of partial rompA sequences demonstrated that the isolate was genotypically different from other validated species of SFG rickettsiae. R. monacensis also replicated in cell lines derived from the ticks I. ricinus (IRE11) and Dermacentor andersoni (DAE100) and in the mammalian cell lines L-929 and Vero, causing cell lysis. Transmission electron microscopy of infected ISE6 and Vero cells showed rickettsiae within the cytoplasm, pseudopodia, nuclei, and vacuoles. Hamsters inoculated with R. monacensis had immunoglobulin G antibody titers as high as 1:16,384, as determined by indirect immunofluorescence assay. Western blot analyses demonstrated that the hamster sera cross-reacted with peptides from other phylogenetically distinct rickettsiae, including rOmpA. R. monacensis induced actin tails in both tick and mammalian cells similar to those reported for R. rickettsii. R. monacensis joins a growing list of SFG rickettsiae that colonize ticks but whose infectivity and pathogenicity for vertebrates are unknown.     

Interspecific comparison of movement patterns among bitterling species in an agricultural ditch system

Patterns of movement are related to life history and/or species traits. Understanding movement patterns of multiple species sharing the same niche may clarify coexistence mechanisms. Therefore, the movement patterns of three bitterling species, Acheilognathus tabira erythropterus, Acheilognathus melanogaster, and Rhodeus ocellatus ocellatus, were examined between June and November 2009 in an agricultural ditch system associated with Lake Kasumigaura in central Japan. The mark–recapture method was used with a fixed releasing point in an approximately 5.5 km-long ditch. Movement distances of A. t. erythropterus and A. melanogaster were longer than that of R. o. ocellatus, but the direction of movement was similar among the three species. However, R. o. ocellatus moved to locations with a low dissolved oxygen concentration and deeper water. These results suggest that R. o. ocellatus may adapt better to environmental fluctuations, because its tolerance to hypoxia is the highest of the three species. Compared with R. o. ocellatus, A. t. erythropterus and A. melanogaster may move relatively longer distances to search for suitable habitats following an environmental fluctuation.     

Molecular phylogeny and rearrangement of rRNA genes in Rickettsia species.

It has previously been observed that Rickettsia prowazekii has an unusual arrangement of the rRNA genes. In this species, the three rRNA genes, 16S (rrs), 23S (rrl), and 5S (rrf), are not linked in the typical arrangements for bacteria. Rather, the 16S rRNA gene has been separated from the 23S and 5S rRNA gene cluster, and the 23S rRNA gene is preceded by a gene which codes for methionyl-tRNAf(Met) formyltransferase (fmt). In this study, we screened the genus Rickettsia for the fmt-rrl motif in order to examine the phylogenetic depth of this unusual rRNA gene organization. A rearranged operon structure was observed in Rickettsia conorii, Rickettsia parkeri, Rickettsia sibirica, Rickettsia rickettsii, Rickettsia amblyomii, Rickettsia montana, Rickettsia rhipicephali, Rickettsia australis, Rickettsia akari, Rickettsia felis, Rickettsia canada, and Rickettsia typhi. There is also evidence for a divided operon in Rickettsia belli, but in this species, the fmt gene could not be identified upstream of the 23S rRNA gene. In order to place the rearrangement event in the evolutionary history of the Rickettsia, phylogenetic analyses were performed based on the fmt-rrl spacer regions and the 23S rRNA genes. Based on these phylogenies, we suggest that the genomic rearrangement of the rRNA genes preceded the divergence of the typhus group and the spotted fever group Rickettsia. The unique organization of the 23S rRNA genes provides a simple diagnostic tool for identification of Rickettsia species.