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To investigate local transmission of Orientia tsutsugamushi by chiggers in North Carolina, USA, we tested remnant serum specimens from patients with eschar undergoing testing for suspected tickborne disease. We identified 11 persons with O. tsutsugamushi antibodies, including 4 who were positive by both assays; none had severe clinical manifestations consistent with scrub typhus.

Investigating new avenues of mosquito control is an important area of entomological research. Examining the effects of various compounds on mosquito biology contributes to the foundation of knowledge from which novel control methods can be built. Caffeine, in particular, is a commonly consumed compound that has not been thoroughly studied for its potential in disrupting the natural life cycle of mosquitoes. In this exploratory study, we analyzed caffeine's effect on the blood-feeding behavior, survival, and fecundity of Aedes albopictus Skuse (Diptera: Culicidae) mosquitoes.Two outcomes, blood-feeding behavior and fecundity, were analyzed in the first experiment in which mosquitoes were exposed to caffeine doses ranging from 0.2 to 2.4 mg/ml. We found a negative linear relationship between dose and fecundity, but no significant impact on blood-feeding behavior. Adjustments were made to the experimental design in which mosquitoes were exposed to doses ranging from 2.5 to 20 mg/ml. From this experiment, we found that caffeine negatively affected blood-feeding behavior, survival, and fecundity especially at higher concentrations.These results suggest that caffeine could be a potential target for future mosquito control research.

North Carolina (NC) experiences some of the highest incidence rates of spotted fever rickettsiosis (SFR) and ehrlichiosis in the United States (US). Due to the non-specific nature of clinical symptoms, minimal utilization of molecular methods when appropriate, and limitations of sero-diagnostic methods, accurate case identification and subsequent public health reporting is challenging. Herein we detail the protocol and early enrollment results for an enhanced surveillance project aiming to generate more accurate estimates of tick-borne disease incidence in NC. Secondary outcomes of interest include: (i) increasing the obtainment rate of convalescent samples (ii) defining demographic and socioeconomic, behavioral/knowledge, entomologic, and environmental risk factors for disease, and (iii) describing the spectrum and clinical course of disease among cases of SFR and ehrlichiosis up to 90 days after symptom onset. In addition, we will collect remnant serum to establish a biorepository of well characterized samples that we intend to make available to researchers. Of the 150 participants enrolled, highlighted results include 49.5% of participants reported being exposed in their own home compared to 43.2% being exposed due to work or travel showing the importance of tick control and education. We also reported more confirmed cases of SFR and ehrlichiosis (15 and 20 respectively) where the North Carolina State Health Department only reported 14 and 11 confirmed cases in the entire state in 2022. Findings from the project will be reported in subsequent publications.

Background Ehrlichiosis is a potentially fatal tick-borne disease that can progress to involve the central nervous system (CNS) (i.e., neuro-ehrlichiosis), particularly in cases where diagnosis and treatment are delayed. Despite a six-fold national increase in the incidence of ehrlichiosis over the past 20 years, recent data on the prevalence and manifestations of neuro-ehrlichiosis are lacking. Methods We conducted a retrospective chart review of all patients tested for ehrlichiosis at University of North Carolina Health facilities between 2018 and 2021 and identified patients who met epidemiological criteria for ehrlichiosis as established by the Council of State and Territorial Epidemiologists and employed by the Centers for Disease Control and Prevention. We estimated the prevalence of neurological symptoms and described the spectrum of neurological manifestations in acute ehrlichiosis, documenting select patient cases in more detail in a case series. Results Out of 55 patients with confirmed or probable ehrlichiosis, five patients (9.1%) had neurologic symptoms, which is notably lower than previous estimates. Neurological presentations were highly variable and included confusion, amnesia, seizures, focal neurological deficits mimicking ischemic vascular events, and an isolated cranial nerve palsy, though all patients had unremarkable neuroimaging at time of presentation. All but one patient had risk factors for severe ehrlichiosis (i.e., older age, immunosuppression). Conclusions Neuro-ehrlichiosis may lack unifying patterns in clinical presentation that would otherwise aid in diagnosis. Clinicians should maintain a high index of suspicion for neuro-ehrlichiosis in patients with acute febrile illness, diverse neurological symptoms, and negative neuroimaging in lone star tick endemic regions. Key points Ehrlichiosis can progress to involve the central nervous system but may present without unifying patterns of neurological symptoms. Increased awareness of the broad spectrum of clinical presentations in neuro-ehrlichiosis is imperative to facilitate prompt diagnosis and treatment.

Background Human Monocytic Ehrlichiosis is caused by infection with the bacteria Ehrlichia chaffeensis through the bite of an infected lone star tick ( Amblyomma americanum ). Patients infected with Human Monocytic Ehrlichiosis often present with symptoms including fever, headache, myalgia, and occasionally a macular rash. The presence of other endemic tick-borne diseases with similar symptoms, such as Rocky Mountain Spotted Fever, complicate the diagnosis of Human Monocytic Ehrlichiosis. Case presentation A patient developed a fever, diffuse myalgia, headache, and a non-productive cough 5 days after a fishing trip in late May in central North Carolina. Over the course of the illness the patient’s symptoms worsened, with arthralgia, bilateral lower extremity erythema and edema, and a developing bilateral rash on the palms. With testing that revealed elevated liver enzymes, a potential for recent tick exposure (e.g., fishing trip), presentation during tick season, and the development of a rash, Rocky Mountain Spotted Fever and Human Monocytic Ehrlichiosis were considered. The patient was prescribed a seven-day course of oral doxycycline and cefalexin, which would provide coverage from Rickettsia , Ehrlichia and gram-positive bacteria typically responsible for cellulitis. Many of the patient’s symptoms resolved or improved, although the right shoulder remained painful to active movement. The patient was prescribed another seven-day course of doxycycline due to his perceived incomplete response to the first course. Approximately 5 weeks after symptom onset (D0 + 36), the patient followed up with a provider for convalescent testing and counseling. Convalescent Ehrlichia and Rickettsia serological tests were ordered. The acute Ehrlichia serology and acute Rickettsia serology were originally non-reactive with both titers measured at < 1:64. Convalescent serology, ordered 28 days after the acute sample collection, showed a greater than four-fold increase in the Ehrlichia IgG titer (1:256), satisfying clinical and laboratory case definitions for ehrlichiosis. In follow-up, 3 weeks later (D0 + 57), the patient reported that most of his pain had subsided, though he still occasionally got shooting nerve pain when exercising. Conclusion This case of Human Monocytic Ehrlichiosis in North Carolina exemplifies the need for a knowledge of spatial epidemiological patterns and clinical manifestations in the diagnosis of tick-borne diseases.

Background The Coronavirus disease 2019 (COVID-19) pandemic was marked by an increase in diagnosis and treatment delays for a range of medical conditions. Yet the impact of the pandemic on the management of tick-borne diseases, which frequently manifest as an acute febrile illness similar to COVID-19, has not been well described. Methods In this retrospective cohort study of patients with suspected tick-borne disease attending the University of North Carolina Health facilities, we compared the timeliness of diagnosis and treatment in a “pre-COVID” period (March 2019 to February 2020) and a “post-COVID” period (March 2020 to February 2021). Participants included patients with an ICD-10 diagnosis code of spotted fever group rickettsiosis or ehrlichiosis and a positive Rickettsia rickettsii or Ehrlichia indirect immunofluorescence assay immunoglobulin G antibody test result. Of the 897 patients who had an eligible diagnosis, 240 (26.8%) met the inclusion criteria. The main outcome was time from initial presentation to definitive diagnosis and treatment. Results During the 2-year study period, 126 (52.5%) patients were grouped in the pre-COVID period and 114 (47.5%) were grouped in the post-COVID period; 120 (50.0%) were female; and 139 (57.9%) were aged > 50 years. Comparing the post-COVID to the pre-COVID period, the adjusted odds ratio (aOR) for delay in treatment > 0 days was 1.81 (95% confidence interval [CI] 1.07–3.07, P  = 0.03), and for a treatment delay > 7 days, 1.65 (95% CI 0.94–2.90, P  = 0.08). The odds of a delay in diagnosis were similar for patients in the post- and pre-COVID periods, with an aOR of 1.61 (95% CI 0.96–2.72, P  = 0.07) for delays > 0 days, and aOR of 1.72 (95% CI 0.99–3.00, P  = 0.05) for delays > 7 days. Conclusions The odds of a delay in treatment > 0 days were significantly higher in the post-COVID period than in the pre-COVID period. However, the odds of a delay in treatment > 7 days, or a delay in diagnosis, were similar between these two periods. Shifts in care-seeking, alternative care delivery models and prioritization of COVID-19 may contribute to diminished timeliness of treatment for patients with tick-borne diseases. Graphical Abstract

Knock-down resistance (kdr) mutations in the voltage-gated sodium channel gene of Aedes species mosquitoes are biomarkers for resistance to pyrethroid insecticides. In the United States, few studies have reported kdr mutations among Aedes albopictus (Skuse) (Diptera: Culicidae) populations. In this study, we sought to compare the presence of kdr alleles among Ae. albopictus mosquitoes collected from Fort Bragg and Wake County, North Carolina. We collected 538 Ae. albopictus mosquitoes, including 156 from 4 sites at Fort Bragg, North Carolina and 382 from 15 sites in Wake County, North Carolina to compare the prevalence of kdr mutations. Of those successfully sequenced, we identified 12 (3.0%) mosquitoes with kdr mutations, all of which were attributed to variants at position 1534 within domain 3. All mutations were found in mosquitoes collected at Wake County sites; no mutations were identified in collections from Fort Bragg. There was a focus of mutations observed at the Wake County sites with approximately 92% (11 of 12) of the mosquitoes with the mutation coming from one site, where kdr mutations represented 24.4% (11 of 45) of all mosquitoes collected. We observed highly focal resistance in a suburban area of Raleigh, which may be attributable to peri-domestic mosquito control activities that involve area dispersal of pyrethroid insecticides. More robust surveillance is needed to monitor the emergence and spread of resistance.

Public health surveillance systems likely underestimate the true prevalence and incidence of SARS-CoV-2 infection due to limited access to testing and the high proportion of subclinical infections in community-based settings. This ongoing prospective, observational study aimed to generate accurate estimates of the prevalence and incidence of, and risk factors for, SARS-CoV-2 infection among residents of a central North Carolina county. From this cohort, we collected survey data and nasal swabs every two weeks and venous blood specimens every month. Nasal swabs were tested for the presence of SARS-CoV-2 virus (evidence of active infection), and serum specimens for SARS-CoV-2-specific antibodies (evidence of prior infection). As of June 23, 2021, we have enrolled a total of 153 participants from a county with an estimated 76,285 total residents. The anticipated study duration is at least 24 months, pending the evolution of the pandemic. Study data are being shared on a monthly basis with North Carolina state health authorities and future analyses aim to compare study data to state-wide metrics over time. Overall, the use of a probability-based sampling design and a well-characterized cohort will enable collection of critical data that can be used in planning and policy decisions for North Carolina and may be informative for other states with similar demographic characteristics.

Public health surveillance systems likely underestimate the true prevalence and incidence of SARS-CoV-2 infection due to limited access to testing and the high proportion of subclinical infections in community-based settings. This ongoing prospective, observational study aimed to generate accurate estimates of the prevalence and incidence of, and risk factors for, SARS-CoV-2 infection among residents of a central North Carolina county. From this cohort, we collected survey data and nasal swabs every two weeks and venous blood specimens every month. Nasal swabs were tested for the presence of SARS-CoV-2 virus (evidence of active infection), and serum specimens for SARS-CoV-2-specific antibodies (evidence of prior infection). As of June 23, 2021, we have enrolled a total of 153 participants from a county with an estimated 76,285 total residents. The anticipated study duration is at least 24 months, pending the evolution of the pandemic. Study data are being shared on a monthly basis with North Carolina state health authorities and future analyses aim to compare study data to state-wide metrics over time. Overall, the use of a probability-based sampling design and a well-characterized cohort will enable collection of critical data that can be used in planning and policy decisions for North Carolina and may be informative for other states with similar demographic characteristics.

Investigating new avenues of mosquito control is an important area of entomological research. Examining the effects of various compounds on mosquito biology contributes to the foundation of knowledge from which novel control methods can be built. Caffeine, in particular, is a commonly consumed compound that has not been thoroughly studied for its potential in disrupting the natural life cycle of mosquitoes. In this exploratory study, we analyzed caffeine's effect on the blood-feeding behavior, survival, and fecundity of Aedes albopictus Skuse (Diptera: Culicidae) mosquitoes. Two outcomes, blood-feeding behavior and fecundity, were analyzed in the first experiment in which mosquitoes were exposed to caffeine doses ranging from 0.2 to 2.4 mg/ml. We found a negative linear relationship between dose and fecundity, but no significant impact on blood-feeding behavior. Adjustments were made to the experimental design in which mosquitoes were exposed to doses ranging from 2.5 to 20 mg/ml. From this experiment, we found that caffeine negatively affected blood-feeding behavior, survival, and fecundity especially at higher concentrations. These results suggest that caffeine could be a potential target for future mosquito control research.