Anaplasma Phagocytophilum Research Articles

Contrasting pathogen prevalence between tick and dog populations at Chornobyl

Abstract Background The 1986 disaster at the Chornobyl Nuclear Power Plant released massive amounts of radioactive material into the local environment. In addition to radiation, remediation efforts and abandonment of military-industrial complexes contributed to contamination with heavy metals, organics, pesticides and other toxic chemicals. Numerous studies have evaluated the effects of this contamination on the local ecology. However, few studies have reported the effect of this contamination on vector-borne pathogens and their hosts. In this manuscript, we characterize tick-borne pathogen presence at two sample locations within the Chornobyl Exclusion Zone, one at the Nuclear Power Plant (NPP) and another 16 km away in Chornobyl City (CC). Methods Ticks and whole-blood samples were collected from free-breeding dogs captured at the NPP and CC. Endpoint PCR and quantitative PCR were used to identify tick species and to assess the presence of specific tick-borne pathogens, including Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Babesia spp., Bartonella spp., Francisella tularensis and general Anaplasmataceae. A droplet digital PCR assay was developed for Babesia canis and A. phagocytophilum to evaluate their presence in dogs from the two populations. Pathogen prevalences between the two sample populations were compared by calculating Z-scores. Results Ticks were identified as Ixodes ricinus (n = 102) and Dermacentor reticulatus (n = 4). Overall, 56.9% of I. ricinus ticks were positive for at least one pathogen. A significantly higher prevalence of A. phagocytophilum and B. burgdorferi was found in ticks at the NPP (44.0% and 42.0%, respectively) compared to CC (23.1% and 19.2%, respectively). Babesia spp. (including B. canis and B. caballi) were detected in 8.8% ticks at similar proportions for both populations. Interestingly, we found a significantly lower level of A. phagocytophilum in dogs at the NPP (1.8%) than in dogs at CC (11.7%). In total, 24.3% of dogs were positive for B. canis, evenly distributed across the two populations. Conclusions The results of this study show contrasting pathogen prevalence in both ticks and dogs at the NPP and CC, which may reflect the differential exposures at the two locations. This work adds an important new component to our understanding of the consequences of prolonged exposure to environmental contamination on the wildlife and ecology within the Chornobyl Exclusion Zone. Graphical Abstract

Diversity and Distribution of Bacterial and Parasitic Tick-Borne Pathogens in Armenia, Transcaucasia

Background: Variations in the distribution and prevalence of pathogens in ticks can have significant consequences for human health. Information on these variables in Transcaucasia is scarce, so the aim of our study was to conduct a large-scale study to detect selected tick-borne infectious agents in Armenia. Methods: Overall, 209 adult ticks were collected from different hosts including 4 samples from human clothes. We tested ticks using high-throughput microfluidic single-cell real-time PCR to detect 42 genospecies of pathogens. We used GIS to determine biotic and abiotic factors governing the prevalence of pathogens and applied statistical analyses to test the association between prevalence of pathogens depending on hosts, locality and environment. Results: From 209 samples, 134 were positive to targeted pathogens. Anaplasma phagocytophilum Foggie, 1949 was the most prevalent case (44%). The highest overall prevalence was observed in ticks from sheep (74%), followed by cows (67%) and calves (60%). The highest multiple infection rates were also detected in sheep (40%) and calves (40%) followed by cows (28%). One statistically significant association was found among co-infections (P<0.05). The prevalence of pathogens varied according to locality. The abundance of Anaplasma spp. is significantly correlated with “slope” and “vegetation” factors. Similar patterns were detected for other pathogens. Conclusion: This was the first large-scale survey of multiple tick-borne pathogens in Armenia and Transcaucasia. The results of this study shed light on spatial variations in pathogen infection rate among adult ticks found on hosts and underline a number of environmental determinants of pathogen distribution among ticks.

Analyses of the gut microbial composition of domestic pig louse Haematopinus suis

Haematopinus suis is an obligatory ectoparasite of the domestic pig, serving as a vector of several swine pathogens and posing great threats to the pig industry. The gut microbiome of lice is thought of an important mediator of their healthy physiology. However, there is a great paucity of lice-associated microbial communities’ structure and function. The current study aimed to profile the gut microbiome and to understand the microbial functions of swine lice by metagenomic sequencing and bioinformatics analyses. In total, 102,358 (77.2 %) nonredundant genes were cataloged, by contrast, only a small proportion of genes were assigned to microbial taxa and functional assemblages. Bacteria of known or potential public health significance such as Anaplasma phagocytophilum, Chlamydia trachomatis, Waddlia chondrophila, Bacillus cereus, and Leptotrichia goodfellowii were observed in all samples. The integrated microbial profile further illustrated the evolutionary relevance of endosymbionts and detailed the functional composition, and findings suggested H. suis may acquire adenosylcobalamin by feeding due to an adenosylcobalamin synthesis defect and a lack of complete synthases of endosymbionts. Sucking lice contained fewer functional genes compared with ticks and fleas probably because of the obligate host specificity of parasitic lice. In addition, the genes from the intestines contained encompassed most of the microbial functional genes in sucking lice. A wide range of unknown taxonomic and functional assemblages were discovered, which improves our understanding related to microbial features and physiological activities of sucking lice. In general, this study increases the characterization of the microbiota of lice and offers clues for preventing and controlling lice infestation in swine production in the future.

Prevalence and genetic diversity of Anaplasma phagocytophilum in wild small mammals from western Yunnan province, China.

Anaplasma phagocytophilum (A. phagocytophilum) is an emerging zoonotic pathogen causing human granulocytic anaplasmosis, linked to small mammal reservoirs that harbor various zoonotic pathogens, underscoring their importance in public health and ecology. This study seeks to determine the prevalence of A. phagocytophilum in small mammals using PCR, then sequence and genotype positive samples, and assess infection risk factors. Small mammals were seasonally captured and a nested polymerase chain reaction (nested-PCR) was conducted targeting the 16S rRNA gene on spleen samples to detect A. phagocytophilum infection from three counties in western Yunnan province, China. Positive samples were sequenced and genotyped, revealing genetic diversity and regional clustering of the pathogen. A total of 1,605 small mammals belonging to 30 species, 18 genera, 6 families, 3 orders were captured seasonally and screened in this region, yielding a 0.93% infection rate with A. phagocytophilum (15/1605). Significant variations in infection rates were observed across different species, counties, and habitats. The 16Sr RNA genes of A. phagocytophilum were categorized into two distinct clades, indicating notable genetic diversity. The identification of genetic variants in spleen samples underscores the potential public health risk and the critical importance of the One Health approach in disease surveillance. Our findings emphasize the necessity for continuous monitoring and highlight the value of nested-PCR testing on spleen samples for accurate prevalence assessment.

Nationwide Survey of Vector-Borne Diseases in Rodents and Mites in Korea: Anaplasma, Ehrlichia, and Rickettsia.

Rodents are reservoirs for zoonotic pathogens, making it essential to study both rodents and their ectoparasites. In 2022 and 2023, we investigated the spatial distribution of rodents and their mites across Korea, focusing on three vector-borne diseases (VBDs): Anaplasma, Ehrlichia, and Rickettsia. A total of 835 wild rodents were collected from 16 locations, each consisting of five distinct environmental settings (mountains, waterways, reservoirs, fields, and paddy fields), with 20 traps per setting, totaling 100 Sherman live folding traps per site. Each rodent was identified using a taxonomic key, and post-mortem examinations led to the collection of 7971 mites (498 pools), followed by PCR analysis. Among the rodents, Anaplasma phagocytophilum was detected in 10.3%, Ehrlichia muris in 0.5%, Ehrlichia ruminantium in 0.2%, and Rickettsia raoultii in 2.9%. In mites, A. phagocytophilum was found in 8.8%, E. muris in 0.2%, R. raoultii in 0.2%, R. endosymbiont in 1.6%, and R. australis in 1.2%. This study marks the first detection of E. muris and R. raoultii in Korean rodents and the first global discovery of E. ruminantium in rodents. The detection of multiple pathogens in mites worldwide highlights the importance of continuous VBD monitoring to mitigate public health risks.

Surveillance of tick-borne pathogens present in ticks (Acari: Ixodidae) removed from companion animals in Louisiana, USA.

Current knowledge of tick distribution and tick-borne pathogen presence across Louisiana is limited. Collaborating with veterinarians across the state, ticks removed from companion animals were recovered and assessed for the presence of zoonotic pathogens. A large number of ticks (n = 959) were removed from companion animals and subsequently screened using qPCR for Anaplasma phagocytophilum, Babesia microti, Borrelia burgdorferi, Bartonella henselae, Ehrlichia chaffeensis, and spotted fever group Rickettsia. Five different tick species, Ixodes scapularis (54.5%), Amblyomma americanum (18.4%), Amblyomma maculatum (12.5%), Dermacentor variabilis (11.2%), and Rhipicephalus sanguineus (0.3%) from different regions of Louisiana were collected from October 2018 to July 2019. There were 15 PCR-positive ticks for Rickettsia parkeri (1.6% prevalence), and four ticks were positive for Ehrlichia chaffeensis (0.4% prevalence). This survey identifies ticks and tick-borne pathogens associated with companion animals and areas for future active surveillance.

Molecular study of some vector–borne diseases in cattle raised in western Türkiye

Unfortunately, global warming, especially the global climate crisis, increases the rate of vector–borne infections. Among the causes of this infection are microorganisms in the Rickettsiales Order, which are Gram–negative and small coccobacillus microorganisms that can multiply within host cells and are dependent on their metabolism, in addition to bacterial infections, protozoa such as Babesia spp. and Theileria spp. are transmitted through vectors and cause serious diseases in animals. This study aimed to investigate the presence of some vector–borne bacterial and protozoan microorganisms in blood samples taken from cattle raised in Mugla province, located in the West of Türkiye, and to reveal relevant disease data for the region. In this study, blood samples taken from 100 cattle were examined using molecular methods. While Anaplasma phagocytophilum was detected in 15 blood samples (15%), Anaplasma ovis agent was detected in eight samples (8%). Anaplasma bovis agent (1%) was identified in only one blood sample. In the samples examined within the scope of the study, Ehrlichia and Rickettsia species from bacteria and Theileria spp. and Babesia spp. from parasitic agents could not be detected. Mugla province, located west of Türkiye, has a subtropical dry summer climate, so the probability of infections transmitted through arthropods is high. Since the agents are transmitted through ticks, conducting more studies on vector–borne diseases is essential. This includes mapping the region’s vector ticks and determining and evaluating the tick carrier and disease maps in cattle. The data obtained is thought to help create regional and national vector–borne disease maps.

B-206 Tick-borne disease molecular testing positivity and coinfection incidence in the northeastern united states: 2021-2023

Abstract Background The CDC estimates that Lyme disease affects hundreds of thousands of people each year in the United States. Tick-borne diseases caused by pathogens such as Anaplasma phagocytophilum, Babesia microti, Ehrlichia chaffeensis, and species of Borrelia follow a seasonal pattern due to the life cycle of disease-carrying ticks. In endemic areas, like the northeastern United States, the risk of coinfection by multiple tick-borne disease pathogens is higher and can complicate patient treatment. For example, while Anaplasma and Borrelia can be treated with doxycycline a Babesia coinfection would not be resolved with this medication. In this study, we performed a retrospective analysis of tick-borne disease molecular testing results to monitor positivity rates and incidence of coinfection. Methods This retrospective analysis included whole blood specimens submitted to a regional clinical laboratory in the northeastern United States for tick-borne disease molecular testing. Test results from 2021-2023 were analyzed for positivity rate trends and coinfection incidence. DNA was isolated from patient specimens and tested using laboratory developed real-time Taq polymerase PCR assays. Results Positivity rates for all tick-borne disease pathogens studied followed typical seasonal patterns (Table). Beginning in the early spring, as nymphs ended their dormancy and began to feed, infection rates of tick-borne disease climbed. Infections peaked in mid-summer and remained elevated through late autumn. For Anaplasma, a biphasic pattern was observed, with a smaller peak in late autumn when adult ticks are most active. Coinfections with Anaplasma and Babesia were observed in 136 specimens, while coinfections with Borrelia and Babesia were observed in 145 specimens. Conclusions Positivity rates for tick-borne disease pathogens followed expected seasonal patterns in the northeastern US. The incidence of coinfections involving Babesia microti highlights the importance of properly identifying tick-borne infections given the different treatment regimens indicated.

A-283 Serologic and molecular testing for the diagnosis of Rickettsia, Ehrlicha, and Anaplasma infections in patients in Maryland

Abstract Background Rickettsia, Anaplasma, and Ehrlichia are underrecognized tick-borne pathogens of increasing human importance. As tick habitats are expanding and populations are increasing, human exposure to these pathogens is also increased in recent years. Unfortunately, while treatment is usually simple, diagnosis can be difficult as it requires specialized testing for clinical diagnosis - generally performed only at reference laboratories. Methods To better understand the prevalence, diagnosis, and testing patterns for Rickettisal infections, Anaplasmosis, and Ehrlichiosis in Maryland, a retrospective analysis of all serologic and molecular testing for Rickettsia, Anaplasma, and Ehrlichia species performed for the University of Maryland Medical System from 2018-2022 was conducted. Eight hospitals within the University of Maryland Medical System (Central Maryland and Maryland’s Eastern Shore) diagnose Rickettsia, Anaplasma, and Ehrlichia infections using serologic and molecular testing. Results Between 2018-2022, an average of 413 individuals were tested for Rickettsia, Anaplasma, and Ehrlichia each year. On average 315 serologic tests for Rocky Mountain Spotted Fever (RMSF) group Rickettsia, 195 serologic tests for Ehrlichia chaffeensis (Anaplasma serology is not available), and 165 molecular tests for the detection of Anaplasma and Ehrlichia species were ordered each year. Clinical indications for testing included tick bite, fever, chills, severe headaches, muscle aches, and rash. Overall, 17% of individuals tested were positive for IgG antibodies to RMSF group Rickettsia, however only 1% were positive for IgM antibodies. Further, 20% of individuals tested were positive for IgG antibodies to Ehrlichia chaffeensis, while <1% were positive for IgM. Acute and convalescent samples were not tested for any individuals. Molecular evidence of infection was found in 8% of individuals tested. Nineteen precent were positive for Anaplasma phagocytophilum and the remaining 81% were positive for Ehrlichia chaffeensis. Interestingly, only two individuals had positive results from both molecular and serologic testing. Conclusions Results from serologic and molecular testing show that Rickettsia, Anaplasma, and Ehrlichia are important human pathogens with the potential to cause significant morbidity. However, lack of follow-up serology, and laboratory tests that confirm acute infection make definitive diagnosis difficult. Increased awareness of Rickettsial diseases and appropriate diagnostics is needed.

Efficacy of a combination of nucleotides and lactoferrin in maintaining stable or improving the clinical picture and laboratory findings of leishmaniotic dogs: A randomized controlled study

This prospective, randomized, controlled, therapeutic study aimed to evaluate the efficacy of a product containing nucleotides and lactoferrin in maintaining or improving the clinical picture and laboratory findings of canine leishmaniosis (CanL). The safety and tolerance of this combination were also assessed. Forty Leishmania infantum-seropositive dogs, not requiring leishmanicidal and/or leishmaniostatic treatment, were enrolled in the study and randomized into treatment (TG) and placebo (CG) groups. Products A (containing nucleotides and lactoferrin) and B (placebo) were blindly administered to TG and CG, respectively, as palatable tablets at a rate of 1 tablet per 10 kg of weight once every 24 h for 6 months. Following inclusion (T0), dogs were followed up after 3 (T90) and 6 (T180) months. At each time point, for all animals enrolled physical examination and laboratory tests (complete blood count, biochemical panel including C-reactive protein [CRP] and ferritin, and serum protein electrophoresis) were performed. The immunofluorescence antibody test to detect antibodies for L. infantum (T0, T180), Ehrlichia canis (T0, T90, and T180), and Anaplasma phagocytophilum (T0, T90, and T180) was executed. A CanL-dedicated clinical score, using a validated scale from 0 (i.e., absence of clinical signs) to 19, was assigned. Four dogs (n=2 in TG, n=2 in CG) did not complete the study. No statistically significant differences in CanL clinical score were observed between CG and TG at T0, T90 and T180. Both TG and CG showed significant variations in anti-L. infantum antibody titres (p=0.0001 and p=0.004, respectively). In TG, antibody titres decreased in 77.8 %, increased in 5.5 %, and remained stable in 16.7 % of dogs, while in CG, decreased in 27.8 %, increased in 50 %, and remained stable in 22.2 % of dogs. During the study, CRP and ferritin remained stable in TG and significantly increased in CG. At T180, 9 out of 18 dogs (50 %) enrolled in the CG, and 1 out of 18 (5.6 %) enrolled in the TG, developed an active form of leishmaniosis. No side effects were reported in any patient included. In conclusion, a 6-month oral administration of a supplement containing nucleotides and lactoferrin was effective in maintaining a stable clinical score, improving antibody titres and potentially reducing the progression from non-active to active forms in L. infatum seropositive dogs. Furthermore, the product was well-tolerated, easy to administer, and free of side effects.

Ticks and Tick-Borne Pathogens in Popular Recreational Areas in Tallinn, Estonia: The Underestimated Risk of Tick-Borne Diseases.

This study reveals a significant presence of ticks and tick-borne pathogens in urban recreational areas of Tallinn, Estonia. During the period of May-June 2018, 815 Ixodes ticks were collected from an area of 11,200 m2 using the flagging method. Tick density reached up to 18.8 ticks per 100 m2, indicating a high concentration of ticks in these urban green spaces. Pathogen analysis demonstrated that 34% of the collected ticks were infected with at least one pathogen. Specifically, Borrelia burgdorferi s.l., the causative agent of Lyme borreliosis, was detected in 17.4% of the ticks; Rickettsia spp. was detected in 13.5%; Neoehrlichia mikurensis was detected in 5.5%; Borrelia miyamotoi was detected in 2.6%; and Anaplasma phagocytophilum and tick-borne encephalitis virus were detected in 0.5% each. These findings indicate that the prevalence and abundance of ticks and tick-borne pathogens in these urban environments are comparable to or even exceed those observed in natural endemic areas. Given the increasing incidence of Lyme borreliosis in Central and Northern Europe, the risk of tick bites and subsequent infection in urban recreational sites should not be underestimated. Public health measures, including enhanced awareness and precautionary information, are essential to mitigate the risk of tick-borne diseases in these urban settings.

Seropositivity to tick-borne pathogens in nature management workers in the Netherlands

The incidence of tick-borne infections other than Lyme borreliosis and tick-borne encephalitis is rising in Europe, including the Netherlands. Nature management workers, being highly exposed to ticks, serve as valuable sentinels for seroprevalence studies on tick-borne pathogens (TBPs). This study assessed nature management workers’ seropositivity to TBPs including Anaplasma phagocytophilum, Babesia divergens, B. microti, Borrelia burgdorferi s.l., Rickettsia conorii and R. typhi in the Netherlands. In addition, the study examined coexposure to multiple TBPs and identified risk factors for B. burgdorferi s.l.- and A. phagocytophilum-seropositivity.The study included 525 nature management workers who donated serum and completed a questionnaire. Sera were analysed for exposure to A. phagocytophilum, B. divergens, B. microti, R. conorii and R. typhi using immunofluorescence assays. For B. burgdorferi s.l. antibody detection, the recommended two-tier testing strategy was used. Risk factor analysis was performed using logistic regression modelling.Seropositivity was 30.9 % for B. burgdorferi s.l.; 16.4 % for A. phagocytophilum; 6.5 % for R. conorii; 2.3 % for R. typhi; 4.2 % for B. divergens; and 0.4 % for B. microti. Almost half (49.3 %) of the participants demonstrated seropositivity for one or more pathogens. Risk factors for B. burgdorferi s.l.-seropositivity included being male, increasing age and tick bite frequency. For A. phagocytophilum-seropositivity, increasing age and working in North Holland province were significant risk factors.This study illustrates the exposure to TBPs in the Netherlands, emphasizing the need for ongoing vigilance and international collaborations to better understand and address the growing threat of TBPs in regions with demonstrated environmental TBP circulation.

Rodent Ecology and Etiological Investigation in China: Results from Vector Biology Surveillance - Shandong Province, China, 2012-2022.

Rodents are hosts of a wide range of zoonotic disease pathogens which threaten human health. However, comprehensive investigations of rodent ecology and etiology in Shandong are lacking. Thus, we aimed to analyze rodent ecology and infection with relevant pathogens in Shandong Province, China. Rodent survey data collected from 2012 to 2022 in Shandong Province were used in this study. Rodents captured from 2020 to 2022 were identified to species and tested for pathogens. From 2012 to 2022, 4,145 rodents were captured, with an average capture rate of 0.70%. High capture rates were observed in rural residential areas and other habitats, such as farmland and forestland. Rattus norvegicus (R. norvegicus) was the dominant species, followed by Mus musculus (M. musculus). The regions with the highest capture rates of R. norvegicus were Dongying (0.82%) and Heze (0.63%), while M. musculus was more prevalent in Dongying (0.81%) and Weihai (0.56%). Rodent capture rates were highest between March and September. The positive detection rates of Hantavirus (HV), Leptospira interrogans (L. interrogans), Rickettsia typhi (R. typhi), Anaplasma phagocytophilum (A. phagocytophilum), and Francisella tularensis (F. tularensis) in rodents were 2.58%, 1.10%, 0.94%, 0.16%, and 0.19%, respectively. The rodent capture rate in human habitation environments has trended downward in Shandong Province, with R. norvegicus and M. musculus being the dominant species. Rodent infection risk from HV, L. interrogans, and R. typhi showed seasonal variation. Strengthening rodent surveillance and maintaining a low capture rate of host animals could be pivotal for preventing and controlling relevant rodent-borne diseases in high-risk areas.

The prevalence of pathogens in ticks collected from humans in Belgium, 2021, versus 2017

BackgroundTicks carry a variety of microorganisms, some of which are pathogenic to humans. The human risk of tick-borne diseases depends on, among others, the prevalence of pathogens in ticks biting humans. To follow-up on this prevalence over time, a Belgian study from 2017 was repeated in 2021.MethodsDuring the tick season 2021, citizens were invited to have ticks removed from their skin, send them and fill in a short questionnaire on an existing citizen science platform for the notification of tick bites (TekenNet). Ticks were morphologically identified to species and life stage level and screened using multiplex qPCR targeting, among others, Borrelia burgdorferi (sensu lato), Anaplasma phagocytophilum, Borrelia miyamotoi, Neoehrlichia mikurensis, Babesia spp., Rickettsia helvetica and tick-borne encephalitis virus (TBEV). The same methodology as in 2017 was used.ResultsIn 2021, the same tick species as in 2017 were identified in similar proportions; of 1094 ticks, 98.7% were Ixodes ricinus, 0.8% Ixodes hexagonus and 0.5% Dermacentor reticulatus. A total of 928 nymphs and adults could be screened for the presence of pathogens. Borrelia burgdorferi (s.l.) was detected in 9.9% (95% CI 8.2–12.0%), which is significantly lower than the prevalence of 13.9% (95% CI 12.2–15.7%) in 2017 (P = 0.004). The prevalences of A. phagocytophilum (4.7%; 95% CI 3.5–6.3%) and R. helvetica (13.3%; 95% CI 11.2–15.6%) in 2021 were significantly higher compared to 2017 (1.8%; 95% CI 1.3–2.7% and 6.8%; 95% CI 5.6–8.2% respectively) (P < 0.001 for both). For the other pathogens tested, no statistical differences compared to 2017 were found, with prevalences ranging between 1.5 and 2.9% in 2021. Rickettsia raoultii was again found in D. reticulatus ticks (n = 3/5 in 2021). Similar to 2017, no TBEV was detected in the ticks. Co-infections were found in 5.1% of ticks. When combining co-infection occurrence in 2017 and 2021, a positive correlation was observed between B. burgdorferi (s.l.) and N. mikurensis and B. burgdorferi (s.l.) and B. miyamotoi (P < 0.001 for both).ConclusionsAlthough the 2021 prevalences fell within expectations, differences were found compared to 2017. Further research to understand the explanations behind these differences is needed.Graphical

Збудники інфекційних хвороб людини і тварин та їх розповсюдження в Європі іксодовим кліщем Dermacentor reticulatus

Due to climate change, the possibility of human infective diseases caused by pathogens from the genera Borelia, Anaplasma, Rickettsia, Babesia, Bartonella, Coxiella, etc. is increasing in Europe. Generally, these pathogens are directly related to arthropod vectors, in particular, to Ixodes ticks. The aim of the study was to determine the current trends in the spread of tick-borne infections transmitted by Dermacentor reticulatus ticks in some European countries, including Ukraine, based on the analysis of literature. Over the past 10 years, in the vast majority of countries, including Europe, the situation with the defeat of farm and wild animals, as well as humans, by pathogens of severe infectious diseases from the genera Borrelia, Anaplasma, Rickettsia, Babesia, Theileria, Coxiella, Bartonella, Francisella, etc. has been aggravated. Representatives of these genera lead to severe diseases, such as tick-borne borreliosis, which are accompanied by damage to the central nervous system, joints, skin, and cardiovascular system. Members of the Rickettsiales genus cause a variety of feverish conditions in animals and humans. Species of the genus Babesia cause babesiosis, a disease caused by damage to red blood cells and the appearance of anti-erythrocyte antibodies, and representatives of the genus Toxoplasma cause toxoplasmosis, accompanied by damage to the nervous, lymphatic, and cardiovascular systems. Q fever is caused by species of the genus Coxiella. Severe human diseases are caused by tick-borne encephalitis viruses. Various PCR test systems have been developed and used to detect these pathogens. For example, in Poland, PCR for Borrelia burgdorferi s.l. was performed using the B. burgdorferi s.l. PCR kit (GeneProof, Czech Republic) for in vitro diagnosis. Anaplasma PCR kits (Blirt-DNA Gdańsk, Poland) were used to detect Anaplasma phagocytophilum DNA. For the detection of Babesia spp., a fragment of the 18S rDNA gene encoding a small ribosomal subunit localized in the conserved region V4 was used. To detect Bartonella spp., the tests Hum PCR Bartonella, Hum PCR Coxiella burnetii, and Vet PCR Rickettsia kit (BioIngenTech, Chile), Coxiella burnetii and Rickettsia spp. were used. In the Laboratory for the Study of Ticks and Tick-Borne Infections of Ternopil National Medical University (Ukraine), pathogens were identified using several test systems, namely DNA A. phagocytophilum, / DNA E. muris, / DNA E. chaffeensis, DNA B. burgdorferi s.l., DNA B. myamotoi; DNA Babesia sp., and RNA Ticks borne of encephalitis viruses (MolGen, Italy), which allow real-time determination on anamplifier "ROTOR Gene 6000" by Corbett Research. Studies in many European countries have shown infection of the Dermacentor reticulatus tick with the pathogens B. burgdorferi s.l. in the range from 1.6 to 49.8 %; A. phagocytophilum – 0.1–2.2 %; Babesia sрp. – 9.8–43.8 %; Coxiella spр. – 0.1–0.2%; Francisella spp. – 0.7–95%; Toxoplasma spр. – 3.2–16.7 %; and ticks borne of encephalitis viruses 8.5–16.6 %. Some Ukrainian scientists presented data on D. reticulatus infection, in particular B. burgdorferi s. l. – 10.1–22.1 %; A. phagocytophilum – 3–25.36 %; Babesia spр. – 1–41.2 %; Rickettsia sp. – 28 %; Bartonella spp. – 5 %; Coxiella spр. – 5.1 %, and tick-borne f encephalitis viruses – 11.9–71 %. The spread of many infectious agents is largely correlated with the spread of their vectors, including ticks of the D. reticulatus species. According to many scientists, this species is widespread in Europe, in particular, two metapopulations are currently distinguished as the western one, which includes the territories of France, Belgium, Slovakia, the Czech Republic, the Netherlands, and Germany, and the eastern one, which covers Lithuania, Latvia, Belarus, eastern and central Poland, and west of the Vistula River up to the Ural Mountains. In the Czech Republic, this species is found in all regions, more often in the southeastern and northwestern regions. In the UK, ticks have been found in West Wales, Devon, and Essex. In Romania, D. reticulatus was found in 17 counties, and in Moldova – in the Lower Dniester region. In Ukraine, this species is widespread from the Carpathians and Prykarpattia, Polissya, and Forest-Steppe, to the southeastern regions and Crimea.

Exposure of American Black Bears (Ursus americanus) to Ticks, Tick-Borne Diseases, and Intestinal Parasites in Wisconsin

We surveyed 159 American black bears (Ursus americanus) over a period of three years for the occurrence of ticks, tick-borne diseases, and intestinal parasites in Wisconsin. We collected blood from the bears to test for the presence of antibodies to Borrelia burgdorferi (Lyme disease), Rickettsia rickettsii (Rocky Mountain spotted fever (RMSF)), Babesia, Ehrlichia, Ehrlichia canis, Brucella canis, and Anaplasma phagocytophilum. We also examined scat samples for intestinal parasites. We commonly found the tick Dermacentor variabilis, but also present the first report of Rhipicephalus sanguineus on black bears. We detected antibodies to Lyme disease and RMSF. We detected antibodies to E. canis for the first time in a bear and both antibodies to R. rickettsii and A. phagocytophilum for the first time in a black bear in Wisconsin. No antibodies for Babesia or Br. canis were detected. We found eggs of the intestinal parasite Baylasascaris transfuga as well as a low number of Toxascara leonina and unknown Capillaria species occurrences in the examined feces.

DETECTION OF BABESIA CF. ODOCOILEI, BABESIA CAPREOLI, AND ANAPLASMA PHAGOCYTOPHILUM IN CERVIDS OF THE SCOTTISH HIGHLANDS, UNITED KINGDOM.

Outbreaks of suspected tick-borne disease (redwater fever) have been reported in captive deer of the Scottish Highlands. In this pilot study, polymerase chain reaction and amplicon sequencing were used to detect tick-borne pathogens in opportunistically collected blood and spleen samples from 63 (healthy, n = 44; diseased, n = 19) cervids, and 45 questing and feeding ticks (Ixodes ricinus) from the outbreak sites in 2021-2022. Potentially pathogenic Babesia species were detected in deer but not identified in ticks, Anaplasma phagocytophilum was detected in both deer and ticks, and Borrelia afzelii was detected in ticks but not in deer. Sequencing confirmed Babesia capreoli and Babesia cf. odocoilei parasitemia in clinically healthy red deer (Cervus elaphus), B. capreoli parasitemia in clinically healthy domestic reindeer (Rangifer tarandus tarandus), and two cases of B. cf. odocoilei-associated hemolytic anemia in white-lipped deer (Cervus albirostris), of which one was fatal despite imidocarb treatment. White-lipped deer appear to be highly susceptible to babesiosis caused by B. cf. odocoilei. This investigation highlights the importance of disease surveillance, including molecular diagnostics, for the detection of emerging tick-borne pathogens in managed populations of cervids.

Metagenome reveals the midgut microbial community of Haemaphysalis qinghaiensis ticks collected from yaks and Tibetan sheep

BackgroundHaemaphysalis qinghaiensis is a tick species distributed only in China. Due to its ability to transmit a variety of pathogens, including species of the genera Anaplasma, Rickettsia, Babesia, and Theileria, it seriously endangers livestock husbandry. However, the microbial community of the midgut of H. qinghaiensis females collected from yaks and Tibetan sheep has not yet been characterized using metagenomic sequencing technology.MethodsHaemaphysalis qinghaiensis were collected from the skins of yaks and Tibetan sheep in Gansu Province, China. Genomic DNA was extracted from the midguts and midgut contents of fully engorged H. qinghaiensis females collected from the two hosts. Metagenomic sequencing technology was used to analyze the microbial community of the two groups.ResultsFifty-seven phyla, 483 genera, and 755 species were identified in the two groups of samples. The ticks from the two hosts harbored common and unique microorganisms. At the phylum level, the dominant common phyla were Proteobacteria, Firmicutes, and Mucoromycota. At the genus level, the dominant common genera were Anaplasma, Ehrlichia, and Pseudomonas. At the species level, bacteria including Anaplasma phagocytophilum, Ehrlichia minasensis, and Pseudomonas aeruginosa along with eukaryotes such as Synchytrium endobioticum and Rhizophagus irregularis, and viruses such as the orf virus, Alphadintovirus mayetiola, and Parasteatoda house spider adintovirus were detected in both groups. In addition, the midgut of H. qinghaiensis collected from yaks had unique microbial taxa including two phyla, eight genera, and 23 species. Unique microorganisms in the midgut of H. qinghaiensis collected from Tibetan sheep included two phyla, 14 genera, and 32 species. Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the functional genes of the microbiome of H. qinghaiensis were annotated to six pathways, and the metabolic pathways included 11 metabolic processes, in which the genes involved in carbohydrate metabolism were the most abundant, followed by the genes involved in lipid metabolism.ConclusionsThese findings indicate that most of the microbial species in the collected H. qinghaiensis ticks were the same in both hosts, but there were also slight differences. The analytical data from this study have enhanced our understanding of the midgut microbial composition of H. qinghaiensis collected from different hosts. The database of H. qinghaiensis microbe constructed from this study will lay the foundation for predicting tick-borne diseases. Furthermore, a comprehensive understanding of tick microbiomes will be useful for understanding vector competency and interactions with ticks and midgut microorganisms.Graphical abstract

Effects of Peromyscus spp. (Rodentia: Cricetidae) presence, land use, and ecotone on Ixodes scapularis (Acari: Ixodidae) ecology in an emergent area for tick-borne disease.

As the range of Ixodes scapularis Say expands, host abundance and land use can play important roles in regions where ticks and their associated pathogens are emerging. Small mammal hosts serve as reservoirs of tick-borne pathogens, with Peromyscus leucopus Rafinesque often considered a primary reservoir. A sympatric species Peromyscus maniculatus Wagner is also a competent reservoir and is notoriously difficult to differentiate from P. leucopus. Anthropogenic land use can alter host and habitat availability, potentially changing tick exposure risk. We tested the hypotheses that tick infestation and pathogen prevalence differ between the two Peromyscus spp. and that host-seeking I. scapularis density and pathogen prevalence differ across land use and ecotone gradients. We live trapped small mammals and collected ticks across 3 land-use classifications and ecotones in Maine, an emergent area for tick-borne disease. We tested each small mammal and tick sample for Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. While both Peromyscus spp. serve as hosts for immature ticks, P. leucopus exhibited a higher tick infestation frequency and intensity. We did not detect any significant difference in pathogen infection prevalence between the two species. The density of I. scapularis nymphs and the density of infected nymphs did not differ significantly between land-use types, though did differ across ecotones. We also noted a significant north/south gradient, with higher tick densities and pathogen prevalence at the southern end of the study area. Our study highlights the potential variability in tick density and pathogen prevalence across fine spatial scales within an emerging region for tick-borne disease.

Ticks and Tick-Borne Pathogens: Occurrence and Host Associations over Four Years of Wildlife Surveillance in the Liguria Region (Northwest Italy).

Tick-borne diseases (TBDs) are a considerable public health problem worldwide. The occurrence of Anaplasma spp., Borrelia burgdorferi s.l., Coxiella burnetii, Rickettsia spp., and tick-borne encephalitis virus (TBEv) was investigated via PCR and sequencing in 683 ticks collected from 105 roe deer, 61 wild boars, 49 fallow deer, and 2 chamois, in the Liguria region, northwest Italy, between 2019 and 2022. The ticks were morphologically identified. Four different tick species were found: Ixodes ricinus (66.8% of the collected ticks), Dermacentor marginatus (15.8%), Rhipicephalus sanguineus s.s. (15.7%), and Haemaphysalis punctata (0.9%). Six ticks (0.9%) were only identified as Rhipicephalus spp. Of the 222 pools analyzed, 27.9% were positive. Most pools (n = 58, 26.1% of pools analyzed) were positive for Rickettsia spp., and several species were found: Rickettsia slovaca was the dominant species (15.3%), followed by R. monacensis (8.1%), while R. helvetica (1.8%), R. massiliae (0.5%), and R. raoultii (0.5%) were found only sporadically. Anaplasma phagocytophilum was identified in three pools and B. burgdorferi s.l. in one pool. All samples were negative for C. burnetii and TBEv. Significant associations were found between I. ricinus and roe deer, D. marginatus and wild boar, and between R. monacensis and I. ricinus. The prevalence of Rickettsia spp. differed significantly between tick and host species. This updated picture of tick species and TBPs in wild ungulates in Liguria, where the population of these animals is increasing, shows a widespread presence of potentially zoonotic Rickettsia spp. Continuous monitoring and public information on preventive measures are needed.