Molecular detection of Ehrlichia spp., Anaplasma spp., and Bartonella spp. in dogs treated at a veterinary teaching clinic in Peru.

In recent years, vector-borne diseases have become widespread throughout the world and affect the health of humans and domestic animals. These diseases spread to areas where their primary vectors, fleas and ticks, thrive, particularly in tropical and subtropical climate regions, providing ideal conditions for their proliferation. The growing closeness between people and their pets increases the likelihood of bites from these ectoparasites, which represents a latent zoonotic risk. Therefore, the objective of the study was to determine the presence of Anaplasma spp., Ehrlichia spp., and Bartonella spp. in dogs treated at the Small Animal Clinic of the Faculty of Veterinary Medicine of the Universidad Nacional Mayor de San Marcos, located in Lima, Peru. Blood samples from 214 dogs were molecularly analyzed for hemopathogen detection. The results revealed prevalences of 2.6 % (6/214) for Anaplasma platys, 5.14 % (11/214) for Ehrlichia canis, and 0.46 % (1/214) for Bartonella rochalimae. No statistically significant relationship was found between the animal infection and the age, sex, breed, presence of fleas (Ctenocephalides felis) and ticks (Rhipicephalus sanguineus), and locality. This study reported molecularly for the first time the presence of A. platys, E. canis, and B. rochalimae in dogs from Lima city, and demonstrates the occurrence of zoonotic pathogens in pets treated at the veterinary clinic.

Tick control prevents carcass condemnations in labs caused by Anaplasma ovis.

Ovine anaplasmosis is causing relevant economic losses in Spain due to icteric carcass condemnation in lambs. Anaplasma ovis infection occurs through grazing sheep that transfer ticks to their offspring. This study compared the efficacy of deltamethrin and cypermethrin pour-on treatments for tick control. A total of 250 PCR A. ovis-positive ewes and their offspring were divided into 5 groups. Group A (50 ewes/50 lambs) was kept as an untreated control group. In groups B (50/50) and C (45/93), the lambs were treated with deltamethrin pour-on and cypermethrin pour-on, respectively, one week after birth. In groups D (50/75) and E (51/68), the ewes were treated with cypermethrin pour-on and deltamethrin pour-on one week before the estimated parturition. External parasite assessment and A. ovis PCR were conducted before treatment and at 21 and 42 days post-treatment. Ewes were checked weekly for tick-detection until weaning. Lamb carcasses were examined at the slaughterhouse. Riphicephalus sanguineus sensu lato ticks were found in ewes throughout the study, with only one tick found in a control group lamb. Three lambs tested positive for A. ovis during the trial, with one condemnation at the abattoir due to jaundice. However, no significant differences were observed between treatment groups. Overall, a significant decrease in infected animals and condemned carcasses was observed compared to previous years, suggesting that deltamethrin and cypermethrin prevent A. ovis transmission from dams to lambs. Further studies are needed to confirm synthetic pyrethroids' effectiveness in controlling tick infestation and averting A. ovis transmission to lambs.

The potential risk of human exposure to tick borne infection by Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Babesia microti in selected recreational areas of the Poprad Landscape Park in southern Poland.

INTRODUCTION AND OBJECTIVE: Ticks (Acari:Ixodida) are dangerous ectoparasites and, at the same time, vectors and/or resevoirs of many pathogens, among others Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti. These ethiological agents of Lyme borreliosis, anaplasmosis and babesiosis are transferred to humans mainly by ticks during feeding. The aim of this study was to estimate the potential risk of human exposure to tick borne infection of B. burgdorferi s.l., A. phagocytophilum and B. microti in selected areas of Poprad Landscape Park in southern Poland [PLP]. MATERIAL AND METHODS: Ixodes ricinus ticks were collected from vegetation by the flagging method. Under a stereoscopic microscope, specimens were determined to the species and developmental stage. In total, DNA was isolated from 363 ticks. To detect B. burgdorferi s.l,.two pairs of primers specific to the flagelline gene were used. In turn, to detect A. phagocytophilum and B. microti, two pairs of primers specific to the 16S rRNA gene fragment and 18S rRNA gene fragment were used, respectively. The amplification products were separated electrophoretically in 2% ethidium bromide stained agarose gels, and visualized under ultra violet light. RESULTS: Generally, pathogens were observed in 19.6% of ticks. Borrelia burgdorferi sensu lato was detected in 11.8% of studied ticks. In turn, A. phagocytophlium and B. microti were presented, respectively, in 0.3% and 7.4% of examined I. ricinus. CONCLUSIONS: The study indicated a potentially high risk of human exposure to infection with tick-borne pathogens, mainly B. burgdorferi s.l. and B. microti, in the areas of PLP. In turn, the presence of A. phagocytophilum in lower percentage was shown in the studied ticks.

Detection and prevalence of Anaplasma phagocytophilum in Ixodes ricinus ticks in eastern Poland.

INTRODUCTION: Anaplasma phagocytophilum are tick-borne bacteria affecting both human and animal health. The aim of the study was to examine the prevalence of A. phagocytophilum in questing I. ricinus ticks collected in Lublin Province, eastern Poland. MATERIAL AND METHODS: Ticks were collected by the flagging method. Total DNA from ticks was extracted by boiling in ammonium hydroxide. Detection of A. phagocytophilum was performed by amplifying a fragment of the 16S rDNA gene. RESULTS: Overall, 626 I. ricinus ticks were tested for the presence of A. phagocytophilum DNA. The prevalence of the pathogenic bacteria was 1.28%. The occurrence of A. phagocytophilum among adults was 1.8%, whereas none of the collected I. ricinus nymphs were infected. CONCLUSIONS: The study revealed the presence of A. phagocytophilum in I. ricinus in eastern Poland, which constitutes a potential health risk for residents, tourists, forestry, and agricultural workers.

Molecular analysis of Anaplasma ovis, Theileria ovis and Brucella abortus in adult Ornithodoros lahorensis soft ticks (Acari: Ixodida: Argasidae) isolated from the Xinjiang Uygur Autonomous Region, China.

Introduction: Ticks are obligate blood-feeding arthropods that cause significant economic losses in domestic animal husbandry and threaten public health. However, information about soft ticks (Acari: Argasidae) and tick-borne pathogens in the Xinjiang Uygur Autonomous Region (XUAR) of China is scarce. Material and Methods: In this study, PCR assays and gene sequencing were used to detect and analyse the epidemiological features of Anaplasma ovis, Theileria ovis and Brucella abortus parasitic infections in 366 Ornithodoros lahorensis soft ticks collected from five sampling sites in the XUAR from October 2019 to March 2022. The ticks were identified by morphological and molecular methods as O. lahorensis. The PCR was conducted using primers complementary to the major surface protein 4 (Msp4) gene of A. ovis, the 18S ribosomal RNA (18S rRNA) of T. ovis and the outer membrane protein 22 (Omp22) gene of B. abortus. Results: The overall infection rate was 91/366 (24.9%) for A. ovis, 127/366 (34.7%) for T. ovis and 94/366 (25.6%) for B. abortus. Sequencing analysis indicated that A. ovis Msp4, T. ovis 18S rRNA and B. abortus Omp22 genes from XUAR isolates showed 99.58-100% identity with documented isolates from other countries. Conclusion: This study provides fundamental evidence for the occurrence of A. ovis, T. ovis and B. abortus in O. lahorensis. Therefore, the potential threat of soft ticks to livestock and humans should not be ignored. This study expands the understanding of the existence of tick-borne pathogens in O. lahorensis and is expected to improve the strategies for prevention and control of ticks and tick-borne diseases in China.

Anaplasma phagocytophilum invasin AipA interacts with CD13 to elicit Src kinase signaling that promotes infection.

Host-microbe interactions that facilitate entry into mammalian cells are essential for obligate intracellular bacterial survival and pathogenesis. Anaplasma phagocytophilum is an obligate intracellular bacterium that invades neutrophils to cause granulocytic anaplasmosis. The invasin-receptor pairs and signaling events that induce Anaplasma uptake are inadequately defined. A. phagocytophilum invasion protein A orchestrates entry via residues 9-21 (AipA9-21) engaging an unknown receptor. Yeast two-hybrid screening suggested that AipA binds within C-terminal amino acids 851-967 of CD13 (aminopeptidase N), a multifunctional protein that, when crosslinked, initiates Src kinase and Syk signaling that culminates in endocytosis. Co-immunoprecipitation validated the interaction and confirmed that it requires the AipA N-terminus. CD13 ectopic expression on non-phagocytic cells increased susceptibility to A. phagocytophilum infection. Antibody blocking and enzymatic inhibition experiments found that the microbe exploits CD13 but not its ectopeptidase activity to infect myeloid cells. A. phagocytophilum induces Src and Syk phosphorylation during invasion. Inhibitor treatment established that Src is key for A. phagocytophilum infection, while Syk is dispensable and oriented the pathogen-invoked signaling pathway by showing that Src is activated before Syk. Disrupting the AipA-CD13 interaction with AipA9-21 or CD13781-967 antibody inhibited Src and Syk phosphorylation and also infection. CD13 crosslinking antibody that induces Src and Syk signaling restored infectivity of anti-AipA9-21-treated A. phagocytophilum. The bacterium poorly infected CD13 knockout mice, providing the first demonstration that CD13 is important for microbial infection in vivo. Overall, A. phagocytophilum AipA9-21 binds CD13 to induce Src signaling that mediates uptake into host cells, and CD13 is critical for infection in vivo. IMPORTANCE: Diverse microbes engage CD13 to infect host cells. Yet invasin-CD13 interactions, the signaling they invoke for pathogen entry, and the relevance of CD13 to infection in vivo are underexplored. Dissecting these concepts would advance fundamental understanding of a convergently evolved infection strategy and could have translational benefits. Anaplasma phagocytophilum infects neutrophils to cause granulocytic anaplasmosis, an emerging disease for which there is no vaccine and few therapeutic options. We found that A. phagocytophilum uses its surface protein and recently identified protective immunogen, AipA, to bind CD13 to elicit Src kinase signaling, which is critical for infection. We elucidated the AipA CD13 binding domain, which CD13 region AipA engages, and established that CD13 is key for A. phagocytophilum infection in vivo. Disrupting the AipA-CD13 interaction could be utilized to prevent granulocytic anaplasmosis and offers a model that could be applied to protect against multiple infectious diseases.

Culture and other direct detection methods to diagnose human granulocytic anaplasmosis.

OBJECTIVES: We sought to assess the performance of 3 laboratory tests on blood specimens for direct detection of Anaplasma phagocytophilum, the cause of human granulocytic anaplasmosis (HGA), in patients tested at a single medical institution in New York State. METHODS: Direct tests included microscopic blood smear examination for intragranulocytic inclusions, polymerase chain reaction (PCR), and culture using the HL-60 cell line. The HGA cases testing positive by only 1 direct test were not included, unless HGA was confirmed by acute or convalescent serology using an indirect immunofluorescent assay. RESULTS: From 1997 to 2009, 71 patients with HGA were diagnosed by at least 1 of the 3 direct test methods. For the subgroup of 55 patients who were tested using all 3 methods, culture was positive for 90.9% (50/55) vs 81.8% (45/55) for PCR vs 63.6% (35/55) for blood smear (P =.002). Most cultures (79.3%) were detected as positive within 1 week of incubation. CONCLUSIONS: Although using culture to detect A phagocytophilum is likely not amenable for implementation in most hospital laboratories, in our experience, culture had the highest yield among the direct tests evaluated.

Prevalence of Anaplasma Spp. and Theileria Spp. Antigens and Antibodies in Housed and Grazing Korean Indigenous Cattle.

Background: Infection with tick-borne pathogens (TBPs) causes anemia, jaundice, and growth retardation in cattle. Many studies have conducted antigen (Ag) tests for major TBPs, such as Anaplasma spp. and Theileria spp., in Korean indigenous cattle (KIC); however, few studies have analyzed antibodies (Ab) against these pathogens. Materials and Methods: This study simultaneously tested 15 housed cattle raised indoor for over a year and 67 grazing cattle for Anaplasma spp. and Theileria spp. Ag using polymerase chain reaction analysis and Ab using enzyme-linked immunosorbent assay. Results: The A. phagocytophilum was detected in 3 housed cattle (20.0%) and 30 grazing cattle (44.8%), whereas the T. orientalis was detected in 3 housed cattle (20.0%) and 54 grazing cattle (80.6%). The positivity rates for Anaplasma spp. Ab did not differ significantly between housed and grazing cattle (4 out of 15 [93.3%] and 55 out of 67 [80.0%], respectively). The positivity rates for Theileria spp. Ab were higher in grazing cattle than in housed cattle (21 of 67 [31.3%] and 0 out of 15 [0.0%], respectively) (P < 0.01). No difference was observed between the frequency of grazing and the abundance of Ab against either pathogen. Meanwhile, as a result of comparing the detection of pathogen Ag and Ab, Anaplasma spp. showed the highest proportions of 73.3% and 47.8% in Ag (-)/Ab (+) for housed cattle and grazing cattle, respectively. Theileria spp. showed the highest rates of 80.0% in Ag (-)/Ab (-) for housed cattle and 52.2% in Ag (+)/Ab (-) for grazing cattle. Conclusions: This is the first study to determine the impact of antibody abundance against Anaplasma spp. and Theileria spp. on antigen prevalence in KIC.

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.

Molecular detection of Anaplasma bovis, Candidatus Anaplasma boleense and Rickettsia spp. in ticks infesting small ruminants.

Anaplasma spp. and Rickettsia spp. are intracellular vector-borne pathogens and harbored by a wide range of ticks and vertebrate hosts. Aim of this study was to molecularly characterize Anaplasma spp. and Rickettsia spp. in different ticks collected from livestock hosts in nine districts of Khyber Pakhtunkhwa (KP), Pakistan. In total, 862 ticks were collected from cattle, goats and sheep. Highest tick's infestation was observed on cattle 56.14% (32/57), followed by goats 45.45% (40/88), and sheep 42.05% (45/107). Rhipicephalus microplus (305/862, 35.38%) was predominant species, followed by Haemaphysalis sulcata (243/862, 28.19%), Hyalomma anatolicum (133/862, 15.42%), Haemaphysalis bispinosa (120/862, 13.92%), and Hyalomma kumari (61/862, 7.07%). A subset of 135 ticks were screened for Anaplasma spp. and Rickettsia spp. based on the amplification of partial 16 S rDNA and outer-membrane protein A (ompA) fragments, respectively. In total, 16 ticks (11.85%) were positive for Anaplasma spp. and Rickettsia spp. Obtained 16 S rDNA sequences for Anaplasma spp. detected in Ha. bispinosa and Ha. sulcata showed 99.98% identity with Anaplasma bovis, while other detected in Rh. microplus showed 99.84% identity with Candidatus Anaplasma boleense. Similarly, detected ompA sequence in Ha. sulcata showed 100% identity with Rickettsia sp. and 97.93% with Rickettsia slovaca, and another sequence detected in Rh. microplus showed 100% identity with Candidatus Rickettsia shennongii. In phylogenetic trees, these sequences clustered with corresponding species from Pakistan, China, Turkey, South Korea, South Africa, and Herzegovina. This is the first study reporting detection of A. bovis in Ha. bispinosa and Ha. sulcata, Ca. A. boleense in Rh. microplus collected from goats, and R. slovaca-like in Ha. sulcata. Our results enforce the need for regular surveillance of Rickettsiales in hard ticks infesting livestock in the region.

Detection of tick-borne pathogens in the pangolin tick, Amblyomma javanense, from Vietnam and Laos, including a novel species of Trypanosoma.

Two species of Southeast Asian pangolins (the Chinese pangolin, Manis pentadactyla and the Malayan or Sunda pangolin, Manis javanica) are critically endangered species. Therefore, knowledge on their parasitic infections is very important, especially considering ticks that can transmit which pathogens. In this study, 32 pangolin ticks (Amblyomma javanense), that were collected in Vietnam and Laos, were analyzed with molecular methods for the presence of tick-borne pathogens. Two members of the family Anaplasmataceae were shown to be present in 14 pangolin ticks, i.e., Candidatus Anaplasma pangolinii and an Ehrlichia sp. In three ticks, a single Rickettsia genotype was also detected, and in seven ticks four 18S rRNA sequence variants of a Babesia sp. Most importantly, a novel protozoan agent, tentatively called here Trypanosoma sp. "PAT14" was detected in one A. javanense nymph. These results imply the first molecular finding of any species of Anaplasma, Ehrlichia and Babesia in pangolin ticks from Vietnam and Laos. On the other hand, detection of a new tick-associated Trypanosoma sp. in A. javanense from Southeast Asia is not only important from a taxonomic point of view, but it is also the first finding of any trypanosomes in the genus Amblyomma in Eurasia and adds pangolins to the potential placental mammalian hosts of any trypanosomes.

High serological and molecular prevalence of Ehrlichia canis and other vector-borne pathogens in dogs from Boa Vista Island, Cape Verde.

Despite the high global impacts of canine vector-borne diseases (CVBD) due to their wide distribution and zoonotic potential, the current epidemiological situation of CVBD in many tropical and subtropical regions remains unknown. This study examines the seroprevalence and molecular prevalence of Ehrlichia canis and other pathogens causing CVBDs (Leishmania infantum, Dirofilaria immitis, Babesia spp., Anaplasma spp. and Hepatozoon canis) in dogs living on the island of Boa Vista (Cape Verde Republic). Blood samples and infesting ticks were taken from 150 dogs across the island (stray, shelter, and pet dogs). Serum samples were tested using a rapid immunochromatographic test (Uranotest® Quattro) that detects antibodies against E. canis, L. infantum, Anaplasma spp. and D. immitis antigen. Levels of serum antibodies against E. canis were measured using the immunofluorescence antibody test (IFAT). In addition, tick-borne pathogens in blood samples (Anaplasma spp., Babesia spp., Hepatozoon spp., and Ehrlichia canis) were detected by microscopy observation and/or PCR plus sequencing. The seroprevalence of E. canis was extremely high at 82% (123/150), as revealed by both immunochromatography and IFAT. Most dogs returning a seropositive test result (82.92%; 102/123) had antibody titres > 1:1280 but showed no clinical signs or notable laboratory abnormalities. Of the 123 animals testing seropositive for E. canis, 67 (54.47%) also presented antibodies against Anaplasma spp., and 13 (10.56%) showed the presence of Hepatozoon spp. gamonts in the blood smear. Ehrlichia canis infection was detected in 17.1% (25/146) of dogs tested by direct sequencing of polymerase chain reaction (PCR) products. Co-infections were detected in seven of these dogs: four dogs tested PCR-positive for both E. canis and A. platys, two dogs tested positive for E. canis and Hepatozoon spp., and one dog tested positive for E. canis, A. platys and Hepatozoon spp. Rhipicephalus sanguineus sensu lato was the only tick species found infesting the canine study population. The high prevalence of tick-borne pathogens detected in dogs from Boa Vista Island highlights a need for improved control measures designed to prevent the transmission of these pathogens.

Molecular Screening and Genetic Identification of Anaplasma platys in Brown Dog Tick (Rhipicephalus sanguineus s. l.) Infested on Stray Dogs in Taiwan.

Anaplasma platys is a tick-borne zoonotic pathogen of canines. In this study, the presence of A. platys was screened for in brown dog ticks (Rhipicephalus sanguineus s. l.) infesting stray dogs in Taiwan to determine overall prevalence. This study represents the first instance of genetic identification of A. platys in brown dog ticks in Taiwan. In total, we examined 324 brown dog ticks for A. platys infection by nested polymerase chain reaction assay targeting the 16S ribosomal RNA gene. The general prevalence of A. platys infection was 3.1%, with 3.6%, 4.0%, and 2.1% in nymph, female, and male ticks, respectively. Monthly prevalence of infection was observed from May to September. Genetic relatedness was determined by comparing the sequences of the 16S rRNA gene obtained from six Taiwan strains and seventeen other strains, representing six genospecies of Anaplasma spp. and three outgroups (Ehrlichia canis, Rickettsia rickettsia, and Escherichia coli). All Taiwan specimens were shown to genetically belong to the A. platys group, and could be clearly discriminated from other Anaplasma spp. Genetic similarities revealed a 100% identity match with various A. platys documented in GenBank. This study highlights the epidemiological importance of geographical transmission of A. platys among dogs and the possible risk for human infections in Taiwan.

Diversity of Anaplasmataceae Transmitted by Ticks (Ixodidae) and the First Molecular Evidence of Anaplasma phagocytophilum and Candidatus Anaplasma boleense in Paraguay.

Anaplasmataceae bacteria are emerging infectious agents transmitted by ticks. The aim of this study was to identify the molecular diversity of this bacterial family in ticks and hosts, both domestic and wild, as well as blood meal sources of free-living ticks in northeastern Paraguay. The bacteria were identified using PCR-HRM, a method optimized for this purpose, while the identification of ticks and their blood meal was performed using conventional PCR. All amplified products were subsequently sequenced. The bacteria detected in the blood hosts included Ehrlichia canis, Anaplasma platys, and Anaplasma phagocytophilum, Candidatus Anaplasma boleense, and Wolbachia spp., which had not been previously reported in the country. Free-living and parasitic ticks on dogs (Canis lupus familiaris) and wild armadillos (Dasypus novemcinctus) were collected and identified as Rhipicephalus sanguineus and Amblyomma spp. The species E. canis, A. platys, A. phagocytophilum, and Ca. A. boleense were detected in domestic dog ticks, and E. canis and A. platys were found for the first time in armadillos and free-living ticks. Blood feeding sources detected in free-living ticks were rodents, humans, armadillos and dogs. Results show a high diversity of tick-borne pathogens circulating among domestic and wild animals in the northeastern region of Paraguay.

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.

Molecular prevalence of Anaplasma marginale, Babesia bigemina, and Theileria orientalis and their associations with Mafriwal cattle's age groups.

Objective: This study was designed to determine the molecular prevalence of hemoparasites and their associations with Mafriwal cattle's age groups. Materials and Methods: Blood samples were taken from the coccygeal veins of calves (n = 92), yearlings (n = 95), lactating (n = 90), and dry (n = 94) cows, which were subjected to microscopic and molecular identification of hemoparasites. The prevalence rate was determined based on the proportion of infected samples in the observed samples. Associations between hemoparasitism and different age groups of Mafriwal cattle were determined by the odds ratio and Fisher's exact test. Results: Babesia bigemina was the most prevalent hemoparasite in monospecies infection (20.8%), while the co-infection of Anaplasma marginale and B. bigemina (36.4%) had the highest molecular prevalence. Highly significant associations of hemoparasitism were observed between calves and yearlings (p < 0.001, Odds ratio = 21.340, 95% CI = 3.200-907.871), lactating (p < 0.01, Odds ratio = 6.600, 95% CI = 1.808-36.516), and dry (p < 0.001, Odds ratio = 10.457, 95% CI = 2.363-96.242) cows. Nevertheless, calves and yearlings were 2-4 times more likely to be co-infected with multiple hemoparasite species in comparison to older age groups. Conclusion: Mafriwal cattle were more susceptible to hemoparasitism with advancing age, but the younger calves were more prone to be co-infected with multiple hemoparasite species.

Prevalence of tick-borne bacterial pathogens in Germany-has the situation changed after a decade?

Introduction: Tick-borne pathogens, such as Borreliella spp., Rickettsia spp., and Anaplasma spp., are frequently detected in Germany. They circulate between animals and tick vectors and can cause mild to severe diseases in humans. Knowledge about distribution and prevalence of these pathogens over time is important for risk assessment of human and animal health. Methods: Ixodes ricinus nymphs were collected at different locations in 2009/2010 and 2019 in Germany and analyzed for tick-borne pathogens by real-time PCR and sequencing. Results: Borreliella spp. were detected with a prevalence of 11.96% in 2009/2010 and 13.10% in 2019 with B. afzelii and B. garinii as dominant species. Borrelia miyamotoi was detected in seven ticks and in coinfection with B. afzelii or B. garinii. Rickettsia spp. showed a prevalence of 8.82% in 2009/2010 and 1.68% in 2019 with the exclusive detection of R. helvetica. The prevalence of Anaplasma spp. was 1.00% in 2009/2010 and 7.01% in 2019. A. phagocytophilum was detected in seven tick samples. None of the nymphs were positive for C. burnetii. Discussion: Here, observed changes in prevalence were not significant after a decade but require longitudinal observations including parameters like host species and density, climatic factors to improve our understanding of tick-borne diseases.

Investigation of Tick-Borne Pathogens in Ixodes Ticks from Bosnia and Herzegovina.

Limited information is available regarding the presence of tick-borne pathogens and their distribution within Ixodes species in Bosnia and Herzegovina. This study aimed to identify Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi sensu lato (s.l.) in Ixodes ticks collected from domestic and wild animals and vegetation in different regions across Bosnia and Herzegovina. A total of 7438 adult ticks, including 4526 Ixodes ricinus, Ixodes canisuga, and Ixodes hexagonus, were collected. Real-time PCR screening of 450 pooled I. ricinus samples revealed a 22.1% infection rate with at least one pathogen. Rickettsia spp. (6.3%) were found in ticks from dogs, cats, and goats, Babesia spp. (3.1%) in ticks from dogs and cattle, A. phagocytophilum (8.8%) in ticks from dogs, goats, and cattle, and B. burgdorferi s.l. (3.4%) in ticks from dogs and cats. Mixed infections with B. burgdorferi s.l. and A. phagocytophilum, as well as B. burgdorferi s.l. and Rickettsia spp., were found in two pools of I. ricinus from dogs and cats, respectively. Additionally, co-infection with Rickettsia spp. and A. phagocytophilum was confirmed in three tick pools from dogs and goats. Each tick from these pooled samples was individually retested to confirm the presence of pathogens. In the examined pooled samples of I. canisuga (1) and I. hexagonus (6), none of the tested pathogens were detected. Our findings represent the first detection of Rickettsia spp., Babesia spp., A. phagocytophilum, and B. burgdorferi s.l. in I. ricinus collected from domestic animals and vegetation in Bosnia and Herzegovina. Considering the established infection rates, the detection of tick-borne pathogens in adult ticks collected from domestic animals and vegetation enriches the current knowledge of the presence of tick-borne pathogens at the local, regional, national, and broader levels.

Zoonotic Tick-Borne Pathogens in Ticks from Vegetation and Alpine Ibex (Capra ibex) in the Maritime Alps, Italy.

In the Maritime Alps (northwestern Italy), we collected ticks from vegetation and Alpine ibex (Capra ibex). Ixodes ricinus was the most abundant species in the study area, questing up to 1824 m a.s.l. and infesting 28 out of 72 ibexes. Haemaphysalis punctata, H. sulcata and Dermacentor marginatus were also collected. The abundance of questing ticks significantly decreased with altitude, with beechwoods being the preferred habitat. By PCR, we identified Borrelia burgdorferi s.l. in questing I. ricinus (28.3%; 95%CI: 19.4-38.6) but not in specimens collected from animals. Rickettsia spp. infected both questing (20.6%; 95%CI: 12.9-30.3) and on-host (30.2%; 95%CI: 21.2-40.4) I. ricinus. Anaplasma phagocytophilum was detected in 4.3% (95%CI: 1.2-10.8) of questing I. ricinus and in 45.3% (95%CI: 34.6-56.4) of I. ricinus collected from ibex. Female I. ricinus collected on animals were significantly more infected with A. phagocytophilum than females collected from vegetation (OR = 11.7; 95%CI: 3.8-48.1). By amplifying and sequencing a fragment of the groEL gene, we identified 13 groEL haplotypes, clustering with ecotypes I and II; ecotype I, prevalent in our sample, is considered zoonotic. Our study demonstrates the presence of different tick-borne zoonotic agents in the study area, encompassing a wide altitudinal range, as confirmed by the ticks found on ibex, a typical mountain-dwelling mammal. The results also confirm the altitudinal range expansion of ticks and associated pathogens in the Alps and suggest that Alpine ibex may act as a reservoir for A. phagocytophilum, as do other wild ungulate species.

Host-pathogen associations inferred from bloodmeal analyses of Ixodes scapularis ticks in a low biodiversity setting.

Tick-borne pathogen emergence is dependent on the abundance and distribution of competent hosts in the environment. Ixodes scapularis ticks are generalist feeders, and their pathogen infection prevalence depends on their relative feeding on local competent and non-competent hosts. The ability to determine what host a larval life stage tick fed on can help predict infection prevalence, emergence, and spread of certain tick-borne pathogens and the risks posed to public health. Here, we use a newly developed genomic target-based technique to detect the source of larval bloodmeals by sampling questing nymphs from Block Island, RI, a small island with a depauperate mammalian community. We used previously designed specific assays to target all known hosts on this island and analyzed ticks for four human pathogenic tick-borne pathogens. We determined the highest proportion of larvae fed on avian species (42.34%), white-footed mice (36.94%), and white-tailed deer (20.72%) and occasionally fed on feral cats, rats, and voles, which are in low abundance on Block Island. Additionally, larvae that had fed on white-footed mice were significantly more likely to be infected with Borrelia burgdorferi and Babesia microti, while larvae that had fed on white-footed mice or white-tailed deer were significantly more likely to be infected with, respectively, mouse- and deer-associated genotypes of Anaplasma phagocytophilum. The ability to detect a nymph's larval host allows for a better understanding of tick feeding behavior, host distribution, pathogen prevalence, and zoonotic risks to humans, which can contribute to better tick management strategies. IMPORTANCE: Tick-borne diseases, such as Lyme disease, babesiosis, and anaplasmosis, pose significant public health burdens. Tick bloodmeal analysis provides a noninvasive sampling method to evaluate tick-host associations and combined with a zoonotic pathogen assay, can generate crucial insights into the epidemiology and transmission of tick-borne diseases by identifying potential key maintenance hosts. We investigated the bloodmeals of questing Ixodes scapularis nymphs. We found that avian hosts, white-footed mice, and white-tailed deer fed the majority of larval ticks and differentially contributed to the prevalence of multiple tick-borne pathogens and pathogen genotypes in a low biodiversity island setting. Unraveling the intricate network of host-vector-pathogen interactions will contribute to improving wildlife management and conservation efforts, to developing targeted surveillance, and vector and host control efforts, ultimately reducing the incidence of tick-borne diseases and improving public health.