Ecological Factors Characterizing the Prevalence of Bacterial Tick-Borne Pathogens in Ixodes ricinus Ticks in Pastures and Woodlands

Ecological changes are recognized as an important driver behind the emergence of infectious diseases. The prevalence of infection in ticks depends upon ecological factors that are rarely taken into account simultaneously. Our objective was to investigate the influences of forest fragmentation, vegetation, adult tick hosts, and habitat on the infection prevalence of three tick-borne bacteria, Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Rickettsia sp. of the spotted fever group, in questing Ixodes ricinus ticks, taking into account tick characteristics. Samples of questing nymphs and adults were taken from 61 pastures and neighboring woodlands in central France. The ticks were tested by PCR of pools of nymphs and individual adults. The individual infection prevalence was modeled using multivariate regression. The highest infection prevalences were found in adult females collected in woodland sites for B. burgdorferi sensu lato and A. phagocytophilum (16.1% and 10.7%, respectively) and in pasture sites for Rickettsia sp. (8.7%). The infection prevalence in nymphs was lower than 6%. B. burgdorferi sensu lato was more prevalent in woodlands than in pastures. Forest fragmentation favored B. burgdorferi sensu lato and A. phagocytophilum prevalence in woodlands, and in pastures, the B. burgdorferi sensu lato prevalence was favored by shrubby vegetation. Both results are probably because large amounts of edges or shrubs increase the abundance of small vertebrates as reservoir hosts. The Rickettsia sp. prevalence was maximal on pasture with medium forest fragmentation. Female ticks were more infected by B. burgdorferi sensu lato than males and nymphs in woodland sites, which suggests an interaction between the ticks and the bacteria. This study confirms the complexity of the tick-borne pathogen ecology. The findings support the importance of small vertebrates as reservoir hosts and make a case for further studies in Europe on the link between the composition of the reservoir host community and the infection prevalence in ticks.Ecological modifications are recognized as one of the main forces behind the emergence of infectious diseases (37). As vectors and wildlife are very sensitive to environmental conditions, ecological changes are expected to have a particular impact on the epidemiology of vector-borne diseases and those with a wildlife origin (29, 48). Several studies have highlighted the influence of factors such as climate change and habitat fragmentation on the risk of tick-borne diseases (20, 67). The risk of a tick-borne disease being transmitted to humans or to animals is closely linked to the prevalence of pathogens in ticks questing for hosts (38, 58). In turn, infection prevalence directly depends on the probability of ticks feeding on an infected reservoir host. This probability results from a combination of the intrinsic characteristics of the species involved (e.g., the host species feeding preference of the tick and the ability of the pathogen to infect different host species) and the characteristics of the host community (e.g., the likelihood of contact between ticks and infected reservoir hosts) that vary in time and space. Due to the difficulty of directly assessing the host community, it may be characterized indirectly by studying landscape and habitat features (3, 9). The increased fragmentation of deciduous forests, for example, favors infection prevalence in ticks that are the agents of Lyme disease in the eastern United States because this fragmentation pattern favors the abundance of a highly competent host reservoir, the white-footed mouse (Peromyscus leucopus) (1, 12). However, studies of the effect of habitat fragmentation on different tick-borne pathogens are scarce (25, 40, 67). Most only report on the infection prevalence of pathogens in ticks according to sampling locations, the stage of tick development, and their sex (18, 56); few studies take into account the interplay or simultaneous effects of explanatory environmental factors (2).In Europe, the Ixodes ricinus tick is one of the most important vectors for animal and human pathogens, especially bacteria (21). These include pathogenic species of the complex Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis, the most prevalent vector-borne human disease in Europe (57); Anaplasma phagocytophilum, the agent of human and animal granulocytic anaplasmosis, considered to be an emerging disease both in human and in animals (8, 61); and Rickettsia helvetica of the spotted fever group, known to be responsible for nonspecific fevers in humans (28).Although they share the same tick vector, B. burgdorferi sensu lato, A. phagocytophilum, and R. helvetica have different ecological cycles and transmission patterns which influence the infection prevalence at different stages of a tick''s life. For B. burgdorferi sensu lato, the maintenance cycle of the bacteria depends on immature stages of I. ricinus ticks feeding on infected reservoir hosts, mainly small rodents and birds that feed on the ground (36, 62). For A. phagocytophilum, small mammals and ruminants are reservoir hosts (16, 22, 69). In contrast to the other two pathogens, R. helvetica is transovarially and sexually transmitted in ticks (13, 33). Ticks are thus considered to be a reservoir host for the bacteria. Small rodents are also suspected to be reservoir hosts in Europe (45), while the role of ungulates remains unknown (60).It is increasingly recognized that a better understanding of the variation of the prevalence of pathogens in ticks within regions of endemicity is critical to the rational design and monitoring of control programs (47). Our objective was to run an exploratory analysis to test the influence of a range of factors on variations in the prevalence of B. burgdorferi sensu lato, A. phagocytophilum, and Rickettsia sp. of the spotted fever group in questing I. ricinus ticks. The factors considered were two habitats (pasture and woodland), forest fragmentation metrics, the vegetation around and near the pasture, and adult tick hosts (deer and cattle); and the analysis took into account factors linked to tick characteristics (tick sex, tick stage, and the density of questing nymphs). Consequently, we analyzed ticks collected in the field for the presence of DNA from the three bacteria and ran an exploratory statistical model using multivariate regression.     

Comparative genomics of first available bovine Anaplasma phagocytophilum genome obtained with targeted sequence capture

Background

Anaplasma phagocytophilum is a zoonotic and obligate intracellular bacterium transmitted by ticks. In domestic ruminants, it is the causative agent of tick-borne fever, which causes significant economic losses in Europe. As A. phagocytophilum is difficult to isolate and cultivate, only nine genome sequences have been published to date, none of which originate from a bovine strain.Our goals were to; 1/ develop a sequencing methodology which efficiently circumvents the difficulties associated with A. phagocytophilum isolation and culture; 2/ describe the first genome of a bovine strain; and 3/ compare it with available genomes, in order to both explore key genomic features at the species level, and to identify candidate genes that could be specific to bovine strains.

Results

DNA was extracted from a bovine blood sample infected by A. phagocytophilum. Following a whole genome capture approach, A. phagocytophilum DNA was enriched 197-fold in the sample and then sequenced using Illumina technology. In total, 58.9% of obtained reads corresponded to the A. phagocytophilum genome, covering 85.3% of the HZ genome. Then by performing comparisons with nine previously-sequenced A. phagocytophilum genomes, we determined the core genome of these ten strains. Following analysis, 1281 coding DNA sequences, including 1001 complete sequences, were detected in the A. phagocytophilum bovine genome, of which four appeared to be unique to the bovine isolate. These four coding DNA sequences coded for "hypothetical proteins of unknown function” and require further analysis. We also identified nine proteins common to both European domestic ruminants tested.

Conclusion

Using a whole genome capture approach, we have sequenced the first A. phagocytophilum genome isolated from a cow. To the best of our knowledge, this is the first time that this method has been used to selectively enrich pathogenic bacterial DNA from samples also containing host DNA. The four proteins unique to the A. phagocytophilum bovine genome could be involved in host tropism, therefore their functions need to be explored.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-973) contains supplementary material, which is available to authorized users.     

Age-dependent penetrance and mapping of the locus for juvenile and early-onset open-angle glaucoma on chromosome 1q (GLC1A) in a French family

The GLC1A locus for autosomal dominant primary open-angle glaucoma (POAG) with juvenile onset (before 20 years) has been mapped to chromosome 1q21– q31. Recently, a French-Canadian family was described in which both juvenile-onset and middle-age or early-onset POAG were observed and linked to GLC1A. We now describe a second POAG family with variable age of onset (range 11–51, median 36 years of age). Linkage to GLC1A was established with a maximum lod score of 6.21 at the D1S452 locus. A recombination event in a severely glaucomatous patient restricted the distal boundary of the GLC1A interval proximal to the AFM154xc9 marker. This study strengthens the idea that early-onset POAG may also be determined by the GLC1A genetic region. Received: 6 May 1996 / Revised: 21 June 1996     

Production of human antitetanus antibodies by hybridomas

Peripheral blood lymphocytes from healthy people recently immunized against tetanus toxoid (TT) were fused with human malignant B-cell lines or mouse myeloma cells (X63 Ag8.653) in an attempt to establish stable B cell hybridomas secreting anti TT antibodies. Human-human fusion experiments were not successful. In contrast, the five heterospecific fusion experiments yielded between 60 and 100% of wells that contained growing hybrids. Five of these hybrids repeatedly secreted anti-TT antibodies. One of the hybrids was cloned and secreted 10-20 micrograms/ml of human IgM, lambda anti-TT antibody. Heterospecific hybridization thus appears as an interesting method to obtain human monoclonal antibodies, allowing the study of their properties.     

Identification and localization on chromosome 1 of a gene controlling the occurrence of periinsulitis in NOD diabetic mice

Diabetic NOD (Non Obese Diabetic) mice show early pancreatic infiltration of mononuclear cells around Langerhans islets (periinsulitis). Study of (NOD x C57BL/6) F1 and F2 mice reveals that periinsulitis is constantly associated with a similar infiltration of salivary glands (sialitis) and is controlled by a dominant susceptibility locus. Segregation analysis of periinsulitis and microsatellite DNA markers indicates that the gene controlling periinsulitis maps to chromosome 1, close to the Bcl-2 locus.