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Background: Ehrlichia chaffeensis is responsible for most cases of human ehrlichiosis, an acute febrile tick-borne disease. This clinical entity is more commonly reported in adults from the United States. Therefore, it is of special interest to characterize this disease in children, given that very few cases in children have been reported outside of this country. Case report: We describe the case of a 15-year-old female from northeastern Mexico with a five-day history of myalgias, arthralgias, fever, abdominal pain, rash, and somnolence. The possibility of tick-borne disease was suspected considering that she lived with three tick-infested dogs that had recently died and a neighbor with similar symptoms who deteriorated rapidly and died a week earlier. Ehrlichia spp. was detected in blood samples by polymerase chain reaction. The patient completed a sevenday course of doxycycline and was discharged with complete resolution of symptoms. Conclusions: This case is the first report of ehrlichiosis in a pediatric patient in Mexico, illustrating the importance of considering tick-borne diseases as a differential diagnosis in patients with rash, fever, and altered level of consciousness. This initial clinical presentation may be indistinct from other conditions such as dengue, meningococcemia, and multisystem inflammatory syndrome in children (MIS-C), among others. [ABSTRACT FROM AUTHOR]

Background. In spring and summer, the population of the Baikal region regularly comes into contact with the pathogens transmitted through the bites of ixodid ticks. In the Center for Diagnosis and Prevention of Tick-Borne Infections (Irkutsk, Russian Federation), we annually detect anaplasmas of the Anaplasma phagocytophilum species, as well as Ehrlichia chaffeensis/E. muris in both ixodid ticks and blood samples from people who have been bitten by ticks. At the same time, there are no data in open sources on the incidence of human granulocytic anaplasmosis and human monocytic ehrlichiosis in the Baikal region. Currently, there is very little information on the studies of intensity of the immune response to anaplasmas and ehrlichia in people living in the surveyed area, although this information is critical for assessing the frequency of contacts and the risk of infection of people in a territory endemic for tick-borne infections. The aim. To update information on the presence and prevalence of specific immunoglobulins M and G to A. phagocytophilum and Ehrlichia sp. among the population of the Irkutsk Region affected by tick bites. Materials and methods. In total, 204 samples of blood serum from the residents of the Irkutsk Region who were registered to be bitten by ticks were analyzed for the presence of IgM and IgG to human monocytic ehrlichiosis and human granulocytic anaplasmosis agents. Results. IgG to A. phagocytophilum were found in 9 samples, IgG to E. chaffeensis/E. muris – in 1 sample; no IgM to both pathogens were found in any sample. Conclusions. The results obtained indicate regular infection of the population with anaplasmas and ehrlichia which is a testifies to the existence of active natural foci of human monocytic ehrlichiosis and human granulocytic anaplasmosis in the Baikal region. To clarify the real epidemic role of these infections, a detailed study of the immune status is required both among healthy individuals and among patients with symptoms of an infectious disease.

Introduction. Knowledge about tick-borne disease (TBD) distribution is necessary to improve prevention, whereas detection of human serum IgG antibodies against relevant pathogens is a method for monitoring TBD prevalence in local population. The study objective was to estimate seroprevalence of IgG antibodies against tick-borne encephalitis virus (TBEV), Borrelia burgdorferi sensu lato, Coxiella burnetii, Anaplasma phagocytophilum, and Ehrlichia chaffeensis/E. muris in healthy residents from the five territories of the Northwestern Federal District of the Russian Federation (Arkhangelsk Oblast, Leningrad Oblast, Pskov Oblast, the Republic of Komi and the Republic of Karelia). Materials and methods. In 20172019, a total of 1244 serum samples from healthy residents, not vaccinated against TBDs or other flavivirus-caused infections was studied by ELISA. Results. 21.7% of the sera samples contained IgG antibodies against a single TBD pathogen, whereas 2.1% showed signs of coinfection with two or more pathogens. The most common were IgG antibodies against TBEV (5 territories, 12.2%), followed by Borrelia burgdorferi sensu lato (5 territories, 3.5%), C. burnetii (4 territories, 2.9%), Anaplasma phagoсytophilum (3 territories, 1.6%), E. chaffeensis/E. muris (5 territories, 1.5%). The IgG antibodies were more common in men (55.2%) than in women (44.8%), being found virtually evenly in age-independent manner (from juniors under 18 to seniors over 60). Conclusion. The results of this first comprehensive serosurveillance study in the Northwestern Federal District of the Russian Federation assessing serum IgG antibodies against tick-borne diseases indicate a wide distribution of such pathogens. Moreover, infections caused by C. burnetii, Anaplasma phagocytophilum, and Ehrlichia chaffeensis/E. muris might be highly underdiagnosed.

Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME), is a Gram-negative obligatory intracellular bacterium, which infects and multiplies in human monocytes and macrophages. Host immune cells produce reactive oxygen species (ROS) to eliminate E. chaffeensis upon infection. E. chaffeensis global transcriptional regulator CtrA activates the expression of GshA and GshB to synthesize glutathione (GSH), the most potent natural antioxidant, upon oxidative stress to combat ROS damage. However, the mechanisms exploited by E. chaffeensis to utilize GSH are still unknown. Here, we found that in E. chaffeensis CtrA activated the expression of glutathione S-transferase (GST) upon oxidative stress, and E. chaffeensis GST utilizes GSH to eliminate ROS and confers the oxidative stress resistance to E. chaffeensis. We found that CtrA bound to the promoter regions of 211 genes, including gst, in E. chaffeensis using chromatin immunoprecipitation coupled to deep sequencing (ChIP-seq). Recombinant E. chaffeensis CtrA directly bound to the gst promoter region determined with electrophoretic mobility shift assay (EMSA), and activated the gst expression determined with reporter assay. Recombinant GST showed GSH conjugation activity towards its typical substrate 2,4-dinitrochlorobenzene (CDNB) in vitro and peptide nucleic acid (PNA) transfection of E. chaffeensis, which can knock down the gst transcription level, reduced bacterial survival upon oxidative stress. Our results demonstrate that E. chaffeensis CtrA regulates GSH utilization, which plays a critical role in resistance to oxidative stress, and aid in the development of new therapeutics for HME. [ABSTRACT FROM AUTHOR]

Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME), is a Gram-negative obligatory intracellular bacterium, which infects and multiplies in human monocytes and macrophages. Host immune cells produce reactive oxygen species (ROS) to eliminate E. chaffeensis upon infection. E. chaffeensis global transcriptional regulator CtrA activates the expression of GshA and GshB to synthesize glutathione (GSH), the most potent natural antioxidant, upon oxidative stress to combat ROS damage. However, the mechanisms exploited by E. chaffeensis to utilize GSH are still unknown. Here, we found that in E. chaffeensis CtrA activated the expression of glutathione S-transferase (GST) upon oxidative stress, and E. chaffeensis GST utilizes GSH to eliminate ROS and confers the oxidative stress resistance to E. chaffeensis. We found that CtrA bound to the promoter regions of 211 genes, including gst, in E. chaffeensis using chromatin immunoprecipitation coupled to deep sequencing (ChIP-seq). Recombinant E. chaffeensis CtrA directly bound to the gst promoter region determined with electrophoretic mobility shift assay (EMSA), and activated the gst expression determined with reporter assay. Recombinant GST showed GSH conjugation activity towards its typical substrate 2,4-dinitrochlorobenzene (CDNB) in vitro and peptide nucleic acid (PNA) transfection of E. chaffeensis, which can knock down the gst transcription level, reduced bacterial survival upon oxidative stress. Our results demonstrate that E. chaffeensis CtrA regulates GSH utilization, which plays a critical role in resistance to oxidative stress, and aid in the development of new therapeutics for HME.

A 66-year-old immunocompetent man with preceding travel through the Northeastern United States developed Guillain-Barré syndrome. A broad search for infections revealed intraerythrocytic parasites on blood smear and positive polymerase chain reaction for Babesia microti; elevated IgM/IgG serologies for Ehrlichia chaffeensis; elevated IgM/IgG serologies and qualitative polymerase chain reaction for Epstein-Barr virus; and fecal culture growth of Arcobacter butzleri. In this report, we discuss the known or suspected association of these infectious agents with Guillain-Barré syndrome. This case also highlights the importance, in the setting of endemic exposure, of screening for multiple coinfections that can be transmitted by the same arthropod vector. [ABSTRACT FROM AUTHOR]

Ixodid ticks are carriers of causative agents of infectious diseases dangerous to humans, including tick-borne encephalitis, ixodid tick-borne borrelioses, rickettsioses, human monocytic ehrlichiosis, human granulocytic anaplasmosis, Congo–Crimean hemorrhagic fever, Omsk hemorrhagic fever, Q fever, and tularemia. These natural focal diseases represent a significant medical problem. It is known that an organism's response to the effect of an infectious agent (and, as a consequence, peculiarities of the course and outcome of the disease) depends not only on the characteristics of the causative agent and external factors, but also significantly on the individual characteristics of the host (particularly, human) genome that predetermine the ability of its immune system to suppress the development of infection. There are two main approaches to the search for human genes predetermining the degree of his susceptibility/resistance to infectious agents. First, the analysis of candidate genes that are presumably relevant to the development of this disease based on known or putative functions of their protein products or on previous data on the orthologous genes in model organisms. Second, genome-wide association analysis, in which a large number of genetic markers evenly distributed across the human genome are simultaneously analyzed. The human genetic predisposition to such infectious diseases as AIDS, tuberculosis, malaria, hepatitis C and B, etc., has been intensively studied, and human genetic factors that specifically predetermine the efficiency of protective reactions against these infections have been already identified. As for infections transmitted by ixodid ticks, the genes of human predisposition to them are those far less studied, and the existing data are rather scattered. This review summarizes already-known genetic factors associated with predisposition to different infections transmitted by ixodid ticks in beings human and laboratory animals, as well as approaches to identification of these factors. [ABSTRACT FROM AUTHOR]

Ehrlichia chaffeensis , a small Gram-negative obligatory intracellular bacterium, infects human monocytes or macrophages, and causes human monocytic ehrlichiosis, one of the most prevalent, life-threatening emerging zoonoses. Reactive oxygen species are produced by the host immune cells in response to bacterial infections. The mechanisms exploited by E. chaffeensis to resist oxidative stress have not been comprehensively demonstrated. Here, we found that E. chaffeensis encodes two functional enzymes, GshA and GshB, to synthesize glutathione that confers E. chaffeensis the oxidative stress resistance, and that the expression of gshA and gshB is upregulated by CtrA, a global transcriptional regulator, upon oxidative stress. We found that in E. chaffeensis , the expression of gshA and gshB was upregulated upon oxidative stress using quantitative RT-PCR. Ehrlichia chaffeensis GshA or GshB restored the ability of Pseudomonas aeruginosa GshA or GshB mutant to cope with oxidative stress, respectively. Recombinant E. chaffeensis CtrA directly bound to the promoters of gshA and gshB , determined with electrophoretic mobility shift assay, and activated the expression of gshA and gshB determined with reporter assay. Peptide nucleic acid transfection of E. chaffeensis , which reduced the CtrA protein level, inhibited the oxidative stress-induced upregulation of gshA and gshB. Our findings provide insights into the function and regulation of the two enzymes critical for E. chaffeensis resistance to oxidative stress and may deepen our understanding of E. chaffeensis pathogenesis and adaptation in hosts. [ABSTRACT FROM AUTHOR]

Ehrlichia chaffeensis causes human monocytic ehrlichiosis (HME), which is one of the most prevalent, life-threatening emerging infectious zoonoses. The life cycle of E. chaffeensis includes ticks and mammals, in which E. chaffeensis proteins are expressed differentially contributing to bacterial survival and infection. Among the E. chaffeensis P28-OMP outer membrane proteins, OMP-1B and P28 are predominantly expressed in tick cells and mammalian macrophages, respectively. The mechanisms regulating this differential expression have not been comprehensively studied. Here, we demonstrate that the transcriptional regulators EcxR and Tr1 regulate the differential expression of omp-1B and p28 in E. chaffeensis. Recombinant E. chaffeensis Tr1 bound to the promoters of omp-1B and p28, and transactivated omp-1B and p28 promoter-EGFP fusion constructs in Escherichia coli. The consensus sequence of Tr1 binding motifs was AC/TTATA as determined with DNase I footprint assay. Tr1 showed a higher affinity towards the p28 promoter than the omp-1B promoter as determined with surface plasmon resonance. EcxR activated the tr1 expression in response to a temperature decrease. At 37°C low level of Tr1 activated the p28 expression. At 25°C high level of Tr1 activated the omp-1B expression, while repressing the p28 expression by binding to an additional site upstream of the p28 gene. Our data provide insights into a novel mechanism mediated by Tr1 regulating E. chaffeensis differential gene expression, which may aid in the development of new therapeutics for HME.

Little information is available about human infections by the members of the genus Ehrlichia in Mexico. Only 2 species, Ehrlichia canis and E. chaffensis, are known to cause disease in this country. We report a fatal case of human monocytic ehrlichiosis in Mexico City in a man who was homeless.

Ehrlichia chaffeensis, a small Gram-negative obligatory intracellular bacterium, infects human monocytes or macrophages, and causes human monocytic ehrlichiosis, one of the most prevalent, life-threatening emerging zoonoses. Reactive oxygen species are produced by the host immune cells in response to bacterial infections. The mechanisms exploited by E. chaffeensis to resist oxidative stress have not been comprehensively demonstrated. Here, we found that E. chaffeensis encodes two functional enzymes, GshA and GshB, to synthesize glutathione that confers E. chaffeensis the oxidative stress resistance, and that the expression of gshA and gshB is upregulated by CtrA, a global transcriptional regulator, upon oxidative stress. We found that in E. chaffeensis, the expression of gshA and gshB was upregulated upon oxidative stress using quantitative RT-PCR. Ehrlichia chaffeensis GshA or GshB restored the ability of Pseudomonas aeruginosa GshA or GshB mutant to cope with oxidative stress, respectively. Recombinant E. chaffeensis CtrA directly bound to the promoters of gshA and gshB, determined with electrophoretic mobility shift assay, and activated the expression of gshA and gshB determined with reporter assay. Peptide nucleic acid transfection of E. chaffeensis, which reduced the CtrA protein level, inhibited the oxidative stress-induced upregulation of gshA and gshB. Our findings provide insights into the function and regulation of the two enzymes critical for E. chaffeensis resistance to oxidative stress and may deepen our understanding of E. chaffeensis pathogenesis and adaptation in hosts.

Human monocytic ehrlichiosis is a tickborne disease with a spectrum of presentations ranging from asymptomatic, mild to fatal. Ehrlichiosis can transiently cause white blood cells abnormalities that mimic leukemia/lymphoma and cases have been, on rare occasions, initially mistaken for hematological malignancies. We report a case of Ehrlichia chaffeensis infection suspected to be acute promyelocytic leukemia at presentation, prompting therapy with all-trans-retinoic acid. Physicians should keep tickborne transmitted illnesses on the differential in patients presenting with pancytopenia, especially in endemic areas.

In 2018, an immunosuppressed woman in southern Taiwan had onset of fever, chills, myalgia, malaise, thrombocytopenia, lymphocytopenia, and elevated hepatic transaminases. Investigation revealed infection with Ehrlichia chaffeensis. This autochthonous case of human monocytotropic ehrlichiosis was confirmed by PCR, DNA sequencing, and seroconversion.

Little information is available about human infections by the members of the genus Ehrlichia in Mexico. Only 2 species, Ehrlichia canis and E. chaffensis, are known to cause disease in this country. We report a fatal case of human monocytic ehrlichiosis in Mexico City in a man who was homeless. [ABSTRACT FROM AUTHOR]

ABSTRACT Ehrlichia chaffeensis is an obligatory intracellular bacterium that causes human monocytic ehrlichiosis, an emerging disease transmitted by the Lone Star tick, Amblyomma americanum. E. chaffeensis outer membrane protein entry triggering protein of Ehrlichia (EtpE) is necessary for bacterial entry into human cells. We investigated the role of EtpE in transmission of the bacteria from tick to human cells and whether or not vaccination with EtpE can prevent transmission of ehrlichiae from ticks to mammals. An antiserum against the recombinant C terminus of EtpE (rEtpE-C), which binds a mammalian cell-surface receptor and triggers bacterial entry, significantly inhibited E. chaffeensis transmission from infected tick cells to human monocytes in culture. Each of five specific-pathogen-free dogs were vaccinated with rEtpE-C along with an immunostimulating complex or were sham vaccinated with the complex alone. Dogs vaccinated with rEtpE-C developed high antibody titers against rEtpE-C and produced interferon-γ-secreting cells, as assessed with the ELISpot assay. All 10 dogs were challenged with A. americanum adult ticks infected as nymphs by syringe inoculation with E. chaffeensis. Upon challenge, both the vaccinated and control dogs became infected by day 1 post-tick attachment, but the majority of rEtpE-C-vaccinated dogs rapidly cleared the infection from the bloodstream as soon as day 7, whereas most of sham-vaccinated dogs remained infected at day 35. Peripheral blood leukocytes from vaccinated dogs had significantly elevated interferon-γ mRNA levels and secreted significantly elevated interferon-γ soon after tick attachment. Thus, the EtpE-C vaccine represents the first ehrlichial protein vaccine demonstrated to reduce bacterial infection in mammals upon challenge with infected ticks. IMPORTANCE The incidence of tick-borne diseases has risen dramatically in the past two decades and continues to rise. Discovered in 1986 and designated a nationally notifiable disease in 1998 by the Centers for Disease Control and Prevention, human monocytic ehrlichiosis, which is caused by the bacterium Ehrlichia chaffeensis, is one of the most prevalent, life-threatening, emerging tick-borne zoonoses in the United States. We investigated the role of the E. chaffeensis protein EtpE in transmission of the bacterium from tick to human cells and in vaccinated dogs with EtpE to assess the efficacy of vaccination against E. chaffeensis-infected tick challenge. Our results help fill gaps in our understanding of E. chaffeensis-derived protective antigens that could be used in a candidate vaccine for immunization of humans to counter tick-transmitted ehrlichiosis.

Hemophagocytic Lymphohistiocytosis (HLH) is a fatal, immunologic syndrome characterized by dysregulated tissue inflammation. HLH can be either primary or secondary; with the latter typically resulting from an infection. Diagnosis requires five or more of the following: fever, splenomegaly, cytopenia, hypertriglyceridemia, hemophagocytosis via biopsy, low natural killer (NK) cell activity, elevated ferritin and soluble CD25 level (sCD25). We present a case of HLH related to ehrlichiosis.In order to mount an effective immune response against microbes such as Ehrlichia chaffeensis, the host must have preserved NK cell function. Being that HLH Is characterized as a state of depleted NK cell function, It is crucial to investigate the role NK cell function has in the setting of HLH on the infectivity of Ehrlichia species.

Background. Early detection of tick-borne pathogens in blood samples of infected people prevents the disease before clinical manifestations using antibiotic and immunoglobulin treatment. As far as tick-borne diseases are associated with significant activation of innate immune response, the changes in the cytokine profile soon after a tick bite could be used as an indicator of early tick-borne infection. Aims. The goal of this study was to characterize the production of cytokines during first 2-4 days after a tick bite in asymptomatic people infected with tick-borne pathogens. Materials and methods. The infection of tick-borne encephalitis virus, B. burgdorferi sensu lato, A. phagocytophilum and E. chaffeensis / E. muris was detected in blood of bitten humans using appropriate commercial real time RT-PCR and ELISA assays. Concentration of 14 cytokines in plasma of 89 infected and 45 non-infected people was established using quantitative ELISA kits. Results. Between 2nd and 4th days after tick bite, the prevalence and concentration of certain cytokines in blood of infected people significantly differed from those of the control group. Bacterial pathogens exhibited very similar patterns of cytokine profiles with induction of IL-1a, IL-8 и IFN-a and suppression of SOD, IL-1Ra и IL-17A. For virus infection the increased concentrations of IL-1 a, IL-8, IL-10, IL-17A, IFN-y and SOD were detected. The only suppressed cytokine was IL-1Ra receptor antagonist. Conclusions. The results suggest that evaluation of innate immune response between 2nd and 4th days after the tick bite could be a useful tool for evaluation of the risk of tick-borne disease for the humans bitten by ticks.

Objective of the study is to investigate natural focality of Ixodidae tick borreliosis, granulocytic anaplasmosis, monocytic ehrli-chiosis in humans in the Republic of Tatarstan. Materials and methods. Utilized were the data from tick studies conducted between 2010–2015. Applying immune-enzymatic analysis, investigated were the blood sera from residents (donors) of Kazan and municipal districts of the Republic of Tatarstan for the presence of specific antibodies to borreliosis, ehrlichiosis, and anaplasmosis pathogens. Results and conclusions. For the first time ever, the data on spontaneous carriage of Ehrlichiosis pathogens in Ixodidae ticks have been obtained. The information received is an indicative of the active circulation of Borrelia, Ehrlichia and Anaplasma in the territory of the region and of necessity to expand the research on the “novel” for the Republic nosological forms of natural-focal infections.

The urgent detection of tick-borne pathogens in attached ticks or in human blood followed by the prophylaxis of infection with specific immunoglobulin and/or antibiotics is one of the most reliable measures to prevent human disease. The aim of this work was to evaluate the usability of real-time PCR (RT-PCR) assay in early detection of tick-borne infections. Using commercial RT-PCR assay (Amplisens, Russia) we studied 361 Ixodes persulcatus ticks, 33 Dermacentor sp. ticks and 448 samples of human blood delivered by people attacked by ticks in Irkutsk region during 2013-2014for presence of DNA/RNA of tick-borne encephalitis virus (TBEV), Borrelia burgdorferi sensu lato (B.b.s.l.), Anaplsma phagocytophilum and Ehrlichia chaffeensis/E. muris. The ELISA assay and direct microscopy were used to validate the detection of TBEV and B.b.s.l. respectively. The highest prevalence of tick-borne infections was observed for I. persulcatus ticks - 6,9; 23,8; 6,9; and 11,9 % for TBEV, B.b.s.l., A. phagocytophillum and E. chaffeensis/E. muris respectively. The Dermacentor sp. ticks were less frequently infected with abovementioned pathogens - 3,0; 6,1; 3,0 and 9,1 % respectively. The prevalence of infection in blood samples was lowest and comprised 2,5; 4,2; 2,2 and 4,5 % respectively. Mixed infections were documented in 6,5 % of ticks and 0,7 % of human blood samples. The testing of blood samples in ELISA and testing of ticks in ELISA and direct microscopy corresponded to the results, obtained by RT-PCR. Thus, RT-PCR is valuable and reliable approach for urgent detection and prophylaxis of tick-borne infections.

Human monocytic ehrlichiosis is a febrile illness caused by Ehrlichia chaffeensis, an intracellular bacterium transmitted by ticks. In Mexico, a case of E. chaffeensis infection in an immunocompetent 31-year-old woman without recognized tick bite was fatal. This diagnosis should be considered for patients with fever, leukopenia, thrombocytopenia, and elevated liver enzyme levels.

Thioredoxins are small ubiquitous proteins that participate in a diverse variety of redox reactions via the reversible oxidation of two cysteine thiol groups in a structurally conserved active site. Here, the NMR solution structures of a reduced and oxidized thioredoxin from Ehrlichia chaffeensis (Ec‐Trx, ECH_0218), the etiological agent responsible for human monocytic ehrlichiosis, are described. The overall topology of the calculated structures is similar in both redox states and is similar to those of other thioredoxins: a five‐stranded, mixed β‐sheet (β1–β3–β2–β4–β5) surrounded by four α‐helices. Unlike other thioredoxins studied by NMR in both redox states, the 1H–15N HSQC spectrum of reduced Ec‐Trx was missing eight additional amide cross peaks relative to the spectrum of oxidized Ec‐Trx. These missing amides correspond to residues Cys35–Glu39 in the active‐site‐containing helix (α2) and Ser72–Ile75 in a loop near the active site, and suggest a change in backbone dynamics on the millisecond‐to‐microsecond timescale associated with the breakage of an intramolecular Cys32–Cys35 disulfide bond in a thioredoxin. A consequence of the missing amide resonances is the absence of observable or unambiguous NOEs to provide the distance restraints necessary to define the N‐terminal end of the α‐helix containing the CPGC active site in the reduced state. This region adopts a well defined α‐helical structure in other reported reduced thioredoxin structures, is mostly helical in oxidized Ec‐Trx and CD studies of Ec‐Trx in both redox states suggests there is no significant difference in the secondary structure of the protein. The NMR solution structure of reduced Ec‐Trx illustrates that the absence of canonical structure in a region of a protein may be owing to unfavorable dynamics prohibiting NOE observations or unambiguous NOE assignments. [ABSTRACT FROM AUTHOR]

Microbiological monitoring of the six districts of the Krasnoyarsk Territory was conducted in order to prove the existence of natural foci of human granulocytic anaplasmosis (HGA) and monocytic ehrlichiosis (HME) on its territory. Two-round PCR method was used for the detection of Anaplasma and Ehrlichia DNA. 374 samples from mites were analyzed with two-round PCR for the presence of Anaplasma and Ehrlichia DNA: 341 specimens of Ixodes persulcatus and 33 specimens of Haemaphysalis concinna. Ticks were collected from vegetation on the flag in the 6 districts of the Krasnoyarsk Territory: Yeniseisky, Achynsky, Sharypovsky, Ilansky, Minusinsky and Ermakovsky, belonging to different climate zones and have landscaped, geo-botanical and faunal features. The study showed that 5.6 % of taiga ticks infected with E. muris, 2,1 % with A. phagocytophilum. In 0.3 % of carriers we identified mixtinfected E. muris and A. phagocytophilum. These data indicate presence of HGA and HME foci in the central, western and southern parts of the Krasnoyarsk Territory.

Ehrlichia chaffeensis , the causative agent of human monocytic ehrlichiosis (HME), is a Gram-negative obligatory intracellular bacterium, which infects and multiplies in human monocytes and macrophages. Host immune cells produce reactive oxygen species (ROS) to eliminate E. chaffeensis upon infection. E. chaffeensis global transcriptional regulator CtrA activates the expression of GshA and GshB to synthesize glutathione (GSH), the most potent natural antioxidant, upon oxidative stress to combat ROS damage. However, the mechanisms exploited by E. chaffeensis to utilize GSH are still unknown. Here, we found that in E. chaffeensis CtrA activated the expression of glutathione S-transferase (GST) upon oxidative stress, and E. chaffeensis GST utilizes GSH to eliminate ROS and confers the oxidative stress resistance to E. chaffeensis . We found that CtrA bound to the promoter regions of 211 genes, including gst , in E. chaffeensis using chromatin immunoprecipitation coupled to deep sequencing (ChIP-seq). Recombinant E. chaffeensis CtrA directly bound to the gst promoter region determined with electrophoretic mobility shift assay (EMSA), and activated the gst expression determined with reporter assay. Recombinant GST showed GSH conjugation activity towards its typical substrate 2,4-dinitrochlorobenzene (CDNB) in vitro and peptide nucleic acid (PNA) transfection of E. chaffeensis , which can knock down the gst transcription level, reduced bacterial survival upon oxidative stress. Our results demonstrate that E. chaffeensis CtrA regulates GSH utilization, which plays a critical role in resistance to oxidative stress, and aid in the development of new therapeutics for HME., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Liang, Yan, Zhang, Yang, Li, Jin, Bai, Wu and Cheng.)

Variations in spatio-temporal patterns of Human Monocytic Ehrlichiosis (HME) infection in the state of Kansas, USA were examined and the relationship between HME relative risk and various environmental, climatic and socio-economic variables were evaluated. HME data used in the study was reported to the Kansas Department of Health and Environment between years 2005–2012, and geospatial variables representing the physical environment [National Land cover/Land use, NASA Moderate Resolution Imaging Spectroradiometer (MODIS)], climate [NASA MODIS, Prediction of Worldwide Renewable Energy (POWER)], and socio-economic conditions (US Census Bureau) were derived from publicly available sources. Following univariate screening of candidate variables using logistic regressions, two Bayesian hierarchical models were fit; a partial spatio-temporal model with random effects and a spatio-temporal interaction term, and a second model that included additional covariate terms. The best fitting model revealed that spatio-temporal autocorrelation in Kansas increased steadily from 2005–2012, and identified poverty status, relative humidity, and an interactive factor, ‘diurnal temperature range x mixed forest area’ as significant county-level risk factors for HME. The identification of significant spatio-temporal pattern and new risk factors are important in the context of HME prevention, for future research in the areas of ecology and evolution of HME, and as well as climate change impacts on tick-borne diseases. [ABSTRACT FROM AUTHOR]

Background Donor-derived infections from organ transplantation are rare occurrences with preoperative screening practices. Ehrlichia chaffeensis, a tick-borne illness, transmitted through solid organ transplantation has not been reported previously to our knowledge. We present cases of 2 renal allograft recipients who developed severe E. chaffeensis infection after receipt of organs from a common deceased donor. Methods The 2 renal transplant patients who developed E. chaffeensis infection are reported in case study format with review of the literature. Results Approximately 3 weeks after renal transplantation, both patients developed an acute febrile illness and rapid clinical decline. Recipient A underwent an extensive infectious workup that revealed positive E. chaffeensis DNA from polymerase chain reaction on peripheral blood. Recipient B's clinical team obtained acute and convalescent antibody titers for E. chaffeensis, which demonstrated acute infection. Recipients A and B were treated with doxycycline and tigecycline, respectively, with clinical cure. Conclusions These cases demonstrate that tick-borne pathogens, such as E. chaffeensis, can be transmitted through renal transplantation. E. chaffeensis can be associated with excessive morbidity and mortality, commonly owing to delay in diagnosis and poor response to non-tetracycline antibiotics. In populations with endemic tick-borne illness, donors should be questioned about tick exposure, and appropriate antibiotics can be administered if indicated. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum and Ehrlichia chaffeensis are obligatory intracellular alpha-proteobacteria that infect human leukocytes and cause potentially fatal emerging zoonoses. In the present study, we determined global protein expression profiles of these bacteria cultured in the human promyelocytic leukemia cell line, HL-60. Mass spectrometric (MS) analyses identified a total of 1,212 A. phagocytophilum and 1,021 E. chaffeensis proteins, representing 89.3% and 92.3% of the predicted bacterial proteomes, respectively. Nearly all bacterial proteins (≥ 99%) with known functions were expressed, whereas only approximately 80% of hypothetical proteins were detected in infected human cells. Quantitative MS/MS analyses indicated that highly-expressed proteins in both bacteria included chaperones, enzymes involved in biosynthesis and metabolism, and outer membrane proteins, such as A. phagocytophilum P44 and E. chaffeensis P28/OMP-1. Among 113 A. phagocytophilum p44 paralogous genes, 110 of them were expressed and 88 of them were encoded by pseudogenes. In addition, bacterial infection of HL-60 cells up-regulated the expression of human proteins involved mostly in cytoskeleton components, vesicular trafficking, cell signaling, and energy metabolism, but down-regulated some pattern recognition receptors involved in innate immunity. Our proteomics data represent a comprehensive analysis of A. phagocytophilum and E. chaffeensis proteomes, and provide a quantitative view of human host protein expression profiles regulated by bacterial infection. The availability of these proteomic data will provide new insights into biology and pathogenesis of these obligatory intracellular pathogens.

An immunocompetent animal disease model based on infection with Ehrlichia chaffeensis would facilitate research toward understanding mechanisms responsible for the broad range of clinical signs associated with human monocytic ehrlichiosis (HME). The adaptability of this model for the experimental feeding of tick species and stages and for testing therapies comparable to those for human diseases are additional advantages of large animal models. Herein, we summarize pathology reports for calves that developed fatal disease after experimental inoculation with E. chaffeensis. Elevated liver enzyme levels and lung pathology among these calves corroborated earlier reports of severe HME. Thus, an experimental disease model based on infection of outbred immunocompetent hosts with E. chaffeensis could be within our grasp for the first time. [ABSTRACT FROM AUTHOR]

Human monocytic ehrlichiosis (HME) is a tick-borne disease transmitted during the summer months in the mid-Atlantic, southeastern and south-central United States. A large proportion of patients presenting with ehrlichiosis must be hospitalized because of the severity of their presenting signs, symptoms and lab abnormalities. We report a case of HME presenting with negative serologies and positive DNA PCR for Ehrlichia chaffeensis during the acute illness. The patient was empirically treated with doxycycline before the availability of diagnostic test results and had a rapid recovery. This report summarizes the common findings of ehrlichiosis on presentation, diagnostic strategies, and treatment options. This case emphasizes the importance of considering tick-borne diseases in the differential diagnosis for patients presenting with nonspecific febrile syndromes in endemic areas and using the clinical scenario to determine whether empiric treatment for a tick-borne disease is necessary. Delaying treatment while awaiting confirmatory tests is unnecessary, and may result in a less favorable patient outcome. [ABSTRACT FROM AUTHOR]

We report the case of a kidney transplant recipient who developed Lyme disease, followed by human granulocytic anaplasmosis (HGA) 3 years later. A review of all previously published cases of Lyme disease (3 cases), HGA (5 cases), and human monocytic ehrlichiosis (HME) (5 cases) in transplant recipients is presented. Manifestations of the cases reviewed were similar to those of non-transplant patients. There appeared to be no obvious correlation between immunosuppression and the occurrence of the illness in the transplant recipients. Serologic testing failed to make a diagnosis in 1 patient with HME in the literature and in our patient with HGA, but molecular tests established the diagnosis in both cases. Tandem infection was observed in 1 patient with two episodes of HME 2 years apart. A high index of suspicion for tick-borne illnesses and appropriate prevention measures are needed for transplant patients with epidemiologic risk factors. [ABSTRACT FROM AUTHOR]

Detection of Ehrlichia chaffeensis is necessary to study interactions between the parasite and its vertebrate and invertebrate hosts. The purpose of this study was to develop a sensitive, specific PCR assay for E. chaffeensis based on the outer membrane protein gene, p28. Candidate primer sets were identified and ranked based on annealing scores, similarities to three major p28 sequence clusters, dissimilarity to E. canis p30, an ortholog of p28, and the proximities of flanking primer sequences for nested PCR. The relative sensitivities of five optimized single-step and two nested PCR assays were determined, and the most sensitive assay was found to be a single-step PCR that was as much as 1000-fold more sensitive than a standard 16S rDNA-based nested PCR assay. This p28-based PCR assay amplified the target amplicon from isolates representative of all three major clusters of known p28 sequences, and this assay did not amplify template prepared from either of the two species most closely related to E. chaffeensis, E. canis and E. muris. These results indicate that this sensitive, specific and isolate-universal single-step PCR assay will be a useful tool in characterizing the transmission of this important zoonotic pathogen. [Copyright &y& Elsevier]

Ehrlichia chaffeensis causes human monocytic ehrlichiosis (HME), which is one of the most prevalent, life-threatening emerging infectious zoonoses. The life cycle of E. chaffeensis includes ticks and mammals, in which E. chaffeensis proteins are expressed differentially contributing to bacterial survival and infection. Among the E. chaffeensis P28-OMP outer membrane proteins, OMP-1B and P28 are predominantly expressed in tick cells and mammalian macrophages, respectively. The mechanisms regulating this differential expression have not been comprehensively studied. Here, we demonstrate that the transcriptional regulators EcxR and Tr1 regulate the differential expression of omp-1B and p28 in E. chaffeensis. Recombinant E. chaffeensis Tr1 bound to the promoters of omp-1B and p28, and transactivated omp-1B and p28 promoter-EGFP fusion constructs in Escherichia coli . The consensus sequence of Tr1 binding motifs was A C / T TATA as determined with DNase I footprint assay. Tr1 showed a higher affinity towards the p28 promoter than the omp-1B promoter as determined with surface plasmon resonance. EcxR activated the tr1 expression in response to a temperature decrease. At 37°C low level of Tr1 activated the p28 expression. At 25°C high level of Tr1 activated the omp-1B expression, while repressing the p28 expression by binding to an additional site upstream of the p28 gene. Our data provide insights into a novel mechanism mediated by Tr1 regulating E. chaffeensis differential gene expression, which may aid in the development of new therapeutics for HME.

Ehrlichia chaffeensis is an obligatory intracellular bacterium that causes human monocytic ehrlichiosis, an emerging disease transmitted by the Lone Star tick, Amblyomma americanum . E. chaffeensis outer membrane protein entry triggering protein of Ehrlichia (EtpE) is necessary for bacterial entry into human cells. We investigated the role of EtpE in transmission of the bacteria from tick to human cells and whether or not vaccination with EtpE can prevent transmission of ehrlichiae from ticks to mammals. An antiserum against the recombinant C terminus of EtpE (rEtpE-C), which binds a mammalian cell-surface receptor and triggers bacterial entry, significantly inhibited E. chaffeensis transmission from infected tick cells to human monocytes in culture. Each of five specific-pathogen-free dogs were vaccinated with rEtpE-C along with an immunostimulating complex or were sham vaccinated with the complex alone. Dogs vaccinated with rEtpE-C developed high antibody titers against rEtpE-C and produced interferon-γ-secreting cells, as assessed with the ELISpot assay. All 10 dogs were challenged with A. americanum adult ticks infected as nymphs by syringe inoculation with E. chaffeensis The incidence of tick-borne diseases has risen dramatically in the past two decades and continues to rise. Discovered in 1986 and designated a nationally notifiable disease in 1998 by the Centers for Disease Control and Prevention, human monocytic ehrlichiosis, which is caused by the bacterium IMPORTANCE The incidence of tick-borne diseases has risen dramatically in the past two decades and continues to rise. Discovered in 1986 and designated a nationally notifiable disease in 1998 by the Centers for Disease Control and Prevention, human monocytic ehrlichiosis, which is caused by the bacterium Ehrlichia chaffeensis , is one of the most prevalent, life-threatening, emerging tick-borne zoonoses in the United States. We investigated the role of the E. chaffeensis protein EtpE in transmission of the bacterium from tick to human cells and in vaccinated dogs with EtpE to assess the efficacy of vaccination against E. chaffeensis -infected tick challenge. Our results help fill gaps in our understanding of E. chaffeensis -derived protective antigens that could be used in a candidate vaccine for immunization of humans to counter tick-transmitted ehrlichiosis., (Copyright © 2020 Budachetri et al.)