Current perspectives and difficulties in the design of acaricides and repellents from plant-derived compounds for tick control.

Ticks and tick-borne diseases have gained increasing attention in recent years due to their impact on public health and significant losses in livestock production. The use of synthetic compounds for tick control is becoming problematic, mainly due to the resistance to commercially available products as well as their toxicity. Therefore, new alternative control methods are required. For this purpose, plant-derived extracts may be considered as effective repellents and/or acaricides. The present literature review focuses on studies evaluating the acaricidal and repellent activity of plant-derived extracts and plant secondary metabolites. We also noted recent advances in protein-ligand-docking simulation to examine the possible toxic effect of natural chemical compounds on ticks. In conclusion, plant-derived repellents/acaricides can be effective against ticks, especially in rural areas and livestock farms.

Repellent effects of Chinese cinnamon oil on nymphal ticks of Haemaphysalis longicornis, Rhipicephalus haemaphysaloides, and Hyalomma asiaticum.

The repellent activity of Chinese cinnamon oil (Cinnamomum cassia) on nymphal ticks (Haemaphysalis longicornis Neumann, Rhipicephalus haemaphysaloides Supino, and Hyalomma asiaticum Schulze and Schlottke) was evaluated in a sample Y-tube bioassay. The results were based on the vertical migration of ticks during the host-seek phase and showed a dose-dependent repellent effect of Chinese cinnamon oil on the tested nymphs after 6 h. For H. longicornis, R. haemaphysaloides, and H. asiaticum at the concentrations (vol/vol) of 3, 3, and 1.5%, the repellent percentages over time were 68-97, 69-94, and 69-93%, respectively, which indicated strong repellent activities against ticks, similar to the positive control DEET (N,N-diethyl-3-methylbenzamide). Chinese cinnamon oil exerted the strongest effect on H. asiaticum nymphs. To our knowledge, this is the first study to investigate the repellent effects of Chinese cinnamon oil on ticks. Chinese cinnamon oil has considerable potential and should be developed as a practical tick repellent.

Efficacy of hydroethanolic extract of Randia aculeata seed against the southern cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) on naturally infested cattle under field conditions.

The cattle tick Rhipicephalus (Boophilus) microplus is a major problem of concern for cattle industry in tropical and subtropical areas. Control of cattle tick is based mainly on the use of chemical acaricides, which has contributed to the emerging problem of selection of resistant tick lineages. Plants have been used as an alternative to conventional acaricidal drugs. On the other hand, the acaricidal activity of hydroethanolic extract of Randia aculeata seed (EHRA) has been demonstrated against R. microplus under laboratory conditions. However, the utility of EHRA seed as a potential acaricidal needs to be determined under field conditions. For this reason, the aim of this study was to evaluate the efficacy of the EHRA against R. microplus sprayed on naturally infested calves, determine the effect of the EHRA seed on acetylcholinesterase activity in R. microplus larval and identify the chemical composition of EHRA. Forty-five male calves were divided in three groups and treated with: G1 water; G2 EHRA 20% w/v and G3 coumaphos 0.2% v/v. Acetylcholinesterase (AChE) activity in R. microplus larvae was determined by a colorimetric assay. The chemical composition of EHRA was accessed through HPLC/MS. Significantly fewer ticks were observed after 24 h on the treated group compared to control group. EHRA significantly inhibited in vitro AChE activity in R. microplus at all tested concentrations. Chlorogenic acid, vanillinic acid, p-coumaric acid, caffeic acid. rutin, quercetin, (-)-epicatechin, 4-hydroxybenzoic acid, quercetin, vanillin, 2,4-dimethoxy-6-methylbenzoic acid, scopoletin and ferulic acid were identified in the extract. The results provided new data for the elucidation of the mechanisms of EHRA acaricide action and to further evaluate the use as a new alternative control agent against R. microplus under in vivo conditions.

Acaricidal activity of a 2-methoxy-clovan-9-ol rich fraction from Eugenia copacabanensis (Myrtaceae) extract on Rhipicephalus microplus (Acari: Ixodidae).

Rhipicephalus microplus (Ixodidae, canestrini, 1888) is an invasive ectoparasite of cattle which causes high economic losses in emerging countries such as Brazil. Phytochemical compounds have been tested as an alternative to synthetic acaricides due to potentially lower mammalian toxicity. This study evaluated the acaricidal activity against R. microplus of the 2-methoxy-clovan-9-ol rich fraction obtained from Eugenia copacabanensis Kiaersk.leaves, a little known Myrtaceae species from the Brazilian Atlantic Forest. This fraction was obtained by maceration with methanol, partition with n-hexane and purification by normal-phase column chromatography. GC-MS, 1H and 13C NMR and IR analysis contributed to the identification of a major compound as the sesquiterpene 2-methoxy-clovan-9-ol, reported for the first time for the Myrtaceae family. The fraction was tested against R. microplus unfed larvae and engorged females and a 93% larval mortality was observed at the concentration of 50 mg mL-1. Lower concentrations of the solution tested demonstrated a significant difference in egg mass weight, hatching and control percentage. Experiments with 50.0 mg mL-1 showed significative results, with lower concentration and maximum efficacy for both assays. The IC50 values for unfed larvae and engorged females were 21.76 and 11.13 mg mL-1, respectively. These results were similar to those obtained in other studies with isolated botanical compounds and essential oils. The lower IC50 for engorged females than for unfed R. microplus larvae had also been described for other plant materials, including plants from the same Myrtaceae family. The present result suggested different mechanisms of action of the compound on the reproductive biology of engorged females, improving its effect against egg viability. These results are important for tick control, suggesting that 2-methoxy-clovan-9-ol could be a potential natural acaricidal product against both R. microplus unfed larvae and engorged females.

Novel Francisella-like endosymbiont and Anaplasma species from Amblyomma nodosum hosted by the anteater Tamandua Mexicana in Mexico.

The microbiome represents a complex network among the various members of the community of microorganisms that are associated with a host. The composition of the bacterial community is essential to supplement multiple metabolic pathways that the host lacks, particularly in organisms with blood-sucking habits such as ticks. On the other hand, some endosymbionts showed some competence with potentially pathogenic microorganisms. Francisella-like endosymbionts (FLEs) encompass a group of gamma-proteobacterias that are closely related to Francisella tularensis, but are usually apathogenic, which brings nutrients like vitamin B and other cofactors to the tick. It has been postulated that the main route of transmission of FLE is vertical; however, evidence has accumulated regarding the possible mechanism of horizontal transmission. Despite growing interest in knowledge of endosymbionts in the Neotropical region, the efforts related to the establishment of their inventory for tick communities are concentrated in South and Central America, with an important gap in knowledge in Mesoamerican countries such as Mexico. For this reason, the aim of this work was to evaluate the presence and diversity of endosymbionts in the highly host-specialized tick Amblyomma nodosum collected from the anteater Tamandua mexicana in Mexico. We analysed 36 A. nodosum for the presence of DNA of endosymbiont (Coxiella and Francisella) and pathogenic (Anaplasma, Borrelia, Ehrlichia and Rickettsia) bacteria. The presence of a member of the genus Francisella and Candidatus Anaplasma brasiliensis was demonstrated. Our findings provide information on the composition of A. nodosum's microbiome, increasing the inventory of bacterial species associated with this hard tick on the American continent.

Natural Clerodendrum-derived tick repellent: learning from Nepali culture.

Ticks attaching to ear canals of humans and animals are the cause of otoacariasis, common in rural areas of Nepal. The plant Clerodendrum viscosum is used in multiple indigenous systems of medicine by ethnic communities in the Indo-Nepali-Malaysian region. Visiting the Chitwan National Park, we learned that in indigenous medicine, flower extract of C. viscosum is utilized to treat digestive disorders and extracts from leaves as tick repellent to prevent ticks from invading or to remove them from the ear canal. The objective of our study was to provide support to indigenous medicine by characterizing the in vivo effect of leave extracts on ticks under laboratory conditions and its phytochemical composition. We collected plant parts of C. viscosum (leaves and flowers) and mango (Mangifera indica) leaves at the Chitwan National Park, previously associated with repellent activity to characterize their effect on Ixodes ricinus ticks by in vivo bioassays. A Q-ToF high-resolution analysis (HPLC-ESI-QToF) was conducted to elucidate phenolic compounds with potential repellent activity. Clerodendrum viscosum and M. indica leaf extracts had the highest tick repellent efficacy (%E = 80-100%) with significant differences when compared to C. viscosum flowers extracts (%E = 20-60%) and phosphate-buffered saline. Phytochemicals with tick repellent function as caffeic acid, fumaric acid and p-coumaric acid glucoside were identified in C. viscosum leaf extracts by HPLC-ESI-QToF, but not in non-repellent flower extracts. These results support the Nepali indigenous medicine application of C. viscosum leaf extracts to repel ticks. Additional research is needed for the development of natural and green repellent formulations to reduce the risks associated with ticks resistant to acaricides.

Essential oil Ageratum conyzoides chemotypes and anti-tick activities.

Rhipicephalus (Boophilus) microplus represents a significant obstacle to animal productivity in tropical and subtropical areas, leading to considerable economic losses for the dairy and meat production industries. Essential oils (EO) extracted from Ageratum conyzoides are known to cause death and induce morphogenetic abnormalities in several insect species. This plant, however, presents morphological flower variations, which range from white to purple, associated to different chemotypes. In this context, this study aimed to conduct a novel assessment on the effects of EO extracted from two A. conyzoides chemotypes in the control of the bovine tick R. microplus. The primary constituents of the oil obtained from white flower samples (WFs) were precocene I (80.4 %) and (E)-caryophyllene (14.8 %), while purple flower oil samples (PFs) contained predominantly ß-acoradiene (12.9 %), γ-amorphene (12.3 %), α-pinene (9.9 %), bicyclogermacrene (8.9 %), α-santalene (8.7 %), and andro encecalinol (5.6 %). Interestingly, only the EO chemotype from A. conyzoides PFs displayed acaricidal activity towards R. microplus larvae, with an LC50 of 1.49 mg/mL.

Complicated Japanese Spotted Fever With Meningitis in an Older Patient: A Case Report.

Japanese spotted fever (JSF) poses a significant public health challenge, mainly due to its atypical presentation in specific demographics. This report details a unique case of JSF in an 89-year-old female who was admitted to a rural hospital exhibiting generalized pain and rapid cognitive decline but no rash. Initially misdiagnosed as polymyalgia rheumatica, her condition was complicated by thrombocytopenia and altered mental state, prompting consideration of tick-borne illnesses. Subsequent serological analysis confirmed JSF despite the absence of its hallmark rash. The patient's condition escalated to include bacteremia and aseptic meningitis. Treatment involved a regimen of minocycline and meropenem, along with endoscopic cauterization of a bleeding rectal ulcer. After treatment, the patient showed improvement and was transferred for rehabilitation. This case highlights the criticality of considering JSF in elderly patients within endemic areas, even when classic symptoms like erythema and petechiae are absent. It underscores the necessity for broad diagnostic perspectives, especially in atypical presentations, and the integration of comprehensive care approaches. The involvement of caregivers and relatives in early detection and seeking medical care promptly is crucial. The report illustrates the complexities in diagnosing and managing advanced JSF cases and stresses the importance of early serological testing and adaptive treatment strategies in managing such challenging cases.

Phenotypic and Target-Directed Screening Yields New Acaricidal Alternatives for the Control of Ticks.

Rhipicephalus microplus, the "common cattle tick", is the most important ectoparasite in livestock worldwide due to the economic and health losses it produces. This tick is a vector for pathogens of several tick-borne diseases. In Latin American countries, damages reach approximately USD 500 million annually due to tick infections, as well as tick-borne diseases. Currently, resistant populations for every chemical group of acaricides have been reported, posing a serious problem for tick control. This study aims to find new alternatives for controlling resistant ticks with compounds derived from small synthetic organic molecules and natural origins. Using BME26 embryonic cells, we performed phenotypic screening of 44 natural extracts from 10 Mexican plants used in traditional medicine, and 33 compounds selected from our chemical collection. We found 10 extracts and 13 compounds that inhibited cell growth by 50% at 50 µg/mL and 100 µM, respectively; the dose-response profile of two of them was characterized, and these compounds were assayed in vitro against different life stages of Rhipicephalus microplus. We also performed a target-directed screening of the activity of triosephosphate isomerase, using 86 compounds selected from our chemical collection. In this collection, we found the most potent and selective inhibitor of tick triosephosphate isomerase reported until now. Two other compounds had a potent acaricidal effect in vitro using adults and larvae when compared with other acaricides such as ivermectin and Amitraz. Those compounds were also selective to the ticks compared with the cytotoxicity in mammalian cells like macrophages or bovine spermatozoids. They also had a good toxicological profile, resulting in promising acaricidal compounds for tick control in cattle raising.

The Meat of the Matter: Understanding and Managing Alpha-Gal Syndrome.

Alpha-gal syndrome is an unconventional food allergy, characterized by IgE-mediated hypersensitivity responses to the glycan galactose-alpha-1,3-galactose (alpha-gal) and not to a food-protein. In this review, we discuss how alpha-gal syndrome reframes our current conception of the mechanisms of pathogenesis of food allergy. The development of alpha-gal IgE is associated with tick bites though the possibility of other parasites promoting sensitization to alpha-gal remains. We review the immune cell populations involved in the sensitization and effector phases of alpha-gal syndrome and describe the current understanding of why allergic responses to ingested alpha-gal can be delayed by several hours. We review the foundation of management in alpha-gal syndrome, namely avoidance, but also discuss the use of antihistamines, mast cell stabilizers, and the emerging role of complementary and alternative therapies, biological products, and oral immunotherapy in the management of this condition. Alpha-gal syndrome influences the safety and tolerability of medications and medical devices containing or derived from mammalian products and impacts quality of life well beyond food choices.

Acaricidal Properties of Four Neem Seed Extracts (Azadirachta indica) on the Camel Tick Hyalomma dromedarii (Acari: Ixodidae).

Tick infestation remains one of the major health problems that affect the productivity and comfort of camels. The control of ticks mainly relies on using chemical acaracides. Limited information is available on the potential benefits and activity of various neem extracts on Hyalomma ticks. The present study investigated the acaricidal activity of neem seed extracts at different concentrations against developmental stages of the camel tick Hyalomma dromedarii in comparison to Butox and diazinon. The acaricidal activity of three extracts, namely, hexane extract (HE), methyl chloride extract (MCE), and methanol extract (ME), of neem seeds (Azadirachta indica) were tested at varying concentrations of 5, 10, 15, and 20% on engorged H. dromedarii female ticks at days 1, 3, 5, 7, 12, 16, 20, 28, 37, and 43 after treatment (DPT). Interestingly, results of applying different neem seed extracts to engorged H. dromedarii female ticks showed that the most effective extract was hexane at concentration 20%, causing 100% mortality at 1st day post-application, while methanol extract at 20% and dichloromethane extract at 20% caused the death of all ticks at 28th day posttreatment as compared to Butox® 5.0 and Diazinon-60, which resulted in mortality of all ticks at 3 and 5 DPT, respectively. In addition, no mortality was reported with the application of aqueous extract (AE), which served as the control group. Furthermore, the neem hexane extract exhibited high efficacy against reproductive performance of female ticks, whereas no fertility or oviposition was reported at all of their concentrations. Additionally, no hatchability occurred using all neem extracts, except the aqueous extract, which showing no effect. In the present study, larvae responded more rapidly to the plant extracts, whereas mortality of all larvae was recorded at 24 h after treatment with 5% hexane. Taken together, this study pointed out that the acaricidal effect of hexane extract of neem seeds was more effective and could be economically used for controlling H. dromedarii ticks.

The Scenario of Ticks and Tick-Borne Pathogens of Sheep on a Mediterranean Island.

Ticks and transmitted pathogens constitute a major concern for livestock health/welfare and productivity for the Mediterranean region, often posing an important zoonotic threat. The aim of this study was to investigate the presence, infection intensity, and seasonality of ticks and tick-borne pathogens on the island of Lesvos in Greece, which was selected as a potential hotspot for their circulation. To this end, 101 sheep farms were visited over a tick activity season, and ticks, blood samples, and questionnaire data were collected. Ticks were identified by species, and DNA from both ticks and blood samples was further investigated using the polymerase chain reaction-reverse line blot (PCR-RLB) technique. In 72.3% of the farms, sheep were found to be infected by 9 ixodid species, with Rhipicephalus turanicus being the most common during the spring/early summer period. As regards tick-borne pathogens (TBPs), 84.9% of the animals were found to be infected with at least one pathogen, the most common being genera of Anaplasma and Theileria, alone or in co-infections. To further characterize the Anaplasma species found, selected samples were sequenced, revealing isolates of A. ovis, A. capra, A. marginale, and A. phagocytophilum. Of the 169 female R. turanicus ticks analyzed by PCR-RLB, 89.9% were harboring at least one TBP belonging to the genera Anaplasma, Ehrlichia, Babesia, Theileria, or Rickettsia. Overall, the data presented in this study revealed a high burden of ticks and TBPs in sheep, including zoonotic species, stressing the need for applying effective monitoring and control programs using a more holistic One Health approach.

[Thalamic lesion in tick-borne encephalitis]. / Dvustoronnee porazhenie talamusa pri kleshchevom entsefalite.

Signal enhancement on T2-weighted images of the thalamus on one or both sides by magnetic resonance imaging (MRI) is among the most frequently observed neuroradiological findings in patients with acute tick-borne encephalitis. The article presents a clinical case of a patient who had an encephalo-polyoencephalomyelitic form of tick-borne encephalitis with MRI picture of a typical bilateral thalamic lesion. Such changes according to neuroimaging studies may have a clinical differential-diagnostic value, and the knowledge of these features by neuroradiologists and neurologists will contribute to the correct diagnosis of acute tick-borne encephalitis, especially most severe forms.

Acaricidal effect of Citrullus colocynthis fruit extract on the camel tick Hyalomma dromedarii (Koch, 1844).

The hard tick Hyalomma dromedarii is one of the most injurious ectoparasites affecting camels in Egypt and several other areas of the world. Hyalomma dromedarii engorged females were dipped in alcoholic extract of Citrullus colocynthis fruits at concentrations of 25, 50, 100, 200 and 400 mg/mL and then observed over four weeks following treatment. There was a marked increase in dead females following single treatment with the above concentrations, particularly for the two highest which caused ≥ 90% mortality. In addition, the extract caused rapid blood digestion in ticks as well as obvious decrease in their oviposition (40% oviposition for the highest concentration compared to 100% for the control group), hatchability (40-45% egg hatching across concentrations compared to 78% for the control group) and fertility index (0.32 compared to 0.56 for the control group). As the estimated LC50 and LC95 were recorded 63.84 and 1046.11 mg/mL, respectively, the study also revealed that dipping in 200 mg/mL of C. colocynthis alcoholic extract is sufficient to control H. dromedarii as it recorded >85% acaricide efficacy, so it might be selected to be the effective dose. Moreover, gas chromatography-mass spectrometry (GC-MS) analysis was carried out on alcoholic extract of C. colocynthis fruits and detected its major constituents of decanoic, oleic, linoleic, carbamic and vaccenic acids which have acaricidal activities impacting tick feeding, digestion and survival. The combined effects of alcoholic extract of C. colocynthis fruits indicated by high mortality rate as well as decreased fecundity and reproductive efficacy supports its effectiveness as a promising management agent for the control of H. dromedarii.

Recently Evolved Francisella-Like Endosymbiont Outcompetes an Ancient and Evolutionarily Associated Coxiella-Like Endosymbiont in the Lone Star Tick (Amblyomma americanum) Linked to the Alpha-Gal Syndrome.

Background: Ticks are hematophagous arthropods that transmit various bacterial, viral, and protozoan pathogens of public health significance. The lone star tick (Amblyomma americanum) is an aggressive human-biting tick that transmits bacterial and viral pathogens, and its bites are suspected of eliciting the alpha-gal syndrome, a newly emerged delayed hypersensitivity following consumption of red meat in the United States. While ongoing studies have attempted to investigate the contribution of different tick-inherent factors to the induction of alpha-gal syndrome, an otherwise understudied aspect is the contribution of the tick microbiome and specifically obligate endosymbionts to the establishment of the alpha-gal syndrome in humans. Materials and Methods: Here we utilized a high-throughput metagenomic sequencing approach to cataloging the entire microbial communities residing within different developmental stages and tissues of unfed and blood-fed ticks from laboratory-maintained ticks and three new geographical locations in the United States. The Quantitative Insights Into Microbial Ecology (QIIME2) pipeline was used to perform data analysis and taxonomic classification. Moreover, using a SparCC (Sparse Correlations for Compositional data) network construction model, we investigated potential interactions between members of the microbial communities from laboratory-maintained and field-collected ticks. Results: Overall, Francisellaceae was the most dominant bacteria identified in the microbiome of both laboratory-raised and field-collected Am. americanum across all tissues and developmental stages. Likewise, microbial diversity was seen to be significantly higher in field-collected ticks compared with laboratory-maintained ticks as seen with a higher number of both Operational Taxonomic Units and measures of species richness. Several potential positive and negative correlations were identified from our network analysis. We observed a strong positive correlation between Francisellaceae, Rickettsiaceae, and Midichloriaceae in both developmental stages and tissues from laboratory-maintained ticks, whereas ovarian tissues had a strong positive correlation of bacteria in the family Xanthobacteraceae and Rhizobiaceae. A negative interaction was observed between Coxiellaceae and Francisellaceae in Illinois, and all the bacteria detected from ticks from Delaware were negatively correlated. Conclusion: This study is the first to catalog the microbiome of Am. americanum throughout its developmental stages and different tissue niches and report the potential replacement of Coxiellaceae by Francisellaceae across developmental stages and tissues tested except in ovarian tissues. These unique and significant findings advance our knowledge and open a new avenue of research to further understand the role of tick microbiome in tick-borne diseases and develop a holistic strategy to control alpha-gal syndrome.

First molecular and functional characterisation of ferritin 2 proteins from Ornithodoros argasid ticks.

Ferritins are iron-binding proteins that play critical functions in iron metabolism. Tick ferritins are essential in blood feeding, reproduction, iron transport, and protection of ticks from the iron-mediated oxidative stress during blood feeding and digestion. In ixodids, ferritin 2 (Fer2) is responsible for iron transport into peripheral tissues, it is critically involved in tick reproduction and has been identified as a good candidate antigen to be included in anti-tick vaccines. In argasids, information on the molecular and functional characteristics of ferritins is almost nonexistent. Given the potential of ixodid Fer2 as a vaccine target, the aim of the current study was to characterise the Fer2 orthologues in Ornithodoros erraticus (OEFer2) and O. moubata (OMFer2), including functional analyses by RNAi gene knockdown and the assessment of the protective efficacy of recombinant Fer2 protein in an animal vaccination trials. Characterisation and analysis of the OMFer2 and OEFer2 amino acid sequences showed high similarity to each other, and high similarity to the Fer2 sequences of ixodid species as well, confirming that Fer2 is highly conserved between both tick families and suggesting a similar function in the physiology of both argasid and ixodid ticks. Fer2 gene knockdown in O. moubata reduced egg hatchability rate and the subsequent number of emerging nymphs-1 up to 71%. Conversely, Fer2 gene knockdown in O. erraticus did not affect the treated ticks even though the Fer2 mRNA expression level was reduced by 90%. The recombinant form of O. moubata Fer2 (tOMFer2) was highly immunogenic and induced strong humoral responses when administered to rabbits formulated with Montanide adjuvant. The protective effect of the anti-tOMFer2 response was limited. While in O. erraticus, we did not observe any protective effect, in O. moubata it induced a significant reduction in oviposition without affecting the other parameters analysed. Accordingly, Fer2 seems to be involved in O. moubata embryogenesis. This study provides the first data on the molecular and functional characterisation of Fer2 in soft tick species and paves the way for further studies aimed at unveiling the functional aspects of Fer2 in soft ticks and confirming its potential as a vaccine candidate antigen.

Ethanolic Extracts of Datura innoxia Have Promising Acaricidal Activity against Rhipicephalus microplus as It Blocks the Glutathione S-Transferase Activity of the Target Tick.

Rhipicephalus microplus is a major bovine ectoparasite that negatively impacts the cattle industry. The acaricidal activity of Datura innoxia ethanolic plant extract against R. microplus, compared with trichlorfon, was examined using the adult immersion test (AIT), and larval packet test (LPT). In vitro acaricidal activity of the selected plant extract against R. microplus engorged females was evaluated at different concentrations (2.5, 5, 10, 20, and 40 mg/mL), and was the same for AIT and LPT. It was further supported by in silico molecular docking of D. innoxia's 21 phytochemicals against the R. microplus Glutathione S-transferases (RmGST) protein's three-dimensional (3D) structure predicted by the trRosetta server. The modeled 3D structure was then evaluated and confirmed with PROCHECK, ERRAT, and Verify3D online servers. To predict the binding mechanisms of these compounds, molecular docking was performed using Auto dock Vina software, and molecular dynamic (MD) simulations were used to investigate the protein atom's dynamic motion. D. innoxia has a relatively higher inhibitory effect on oviposition (from 9.81% to 45.37%) and total larval mortality (42.33% at 24 h and 93.67% at 48 h) at 40 mg/mL. Moreover, the docking results showed that the chemicals norapoatropine and 7-Hydroxyhyoscyamine have strong interactions with active site residues of the target protein, with a docking score of -7.3 and -7.0 Kcal/mol, respectively. The current work also provided a computational basis for the inhibitors of Glutathione S-transferases that were studied in this research work, and this new knowledge should aid in creating new and effective acaricidal chemicals. Furthermore, this plant extract's acaricide activity and its effect on oviposition and larval mortality were established in this work for the first time, indicating the possible use of this extract in the management of ticks.

Risk factors for Lyme disease stage and manifestation using electronic health records.

BACKGROUND: Little is known about risk factors for early (e.g., erythema migrans) and disseminated Lyme disease manifestations, such as arthritis, neurological complications, and carditis. No study has used both diagnoses and free text to classify Lyme disease by disease stage and manifestation. METHODS: We identified Lyme disease cases in 2012-2016 in the electronic health record (EHR) of a large, integrated health system in Pennsylvania. We developed a rule-based text-matching algorithm using regular expressions to extract clinical data from free text. Lyme disease cases were then classified by stage and manifestation using data from both diagnoses and free text. Among cases classified by stage, we evaluated individual, community, and health care variables as predictors of disseminated stage (vs. early) disease using Poisson regression models with robust errors. Final models adjusted for sociodemographic factors, receipt of Medical Assistance (i.e., Medicaid, a proxy for low socioeconomic status), primary care contact, setting of diagnosis, season of diagnosis, and urban/rural status. RESULTS: Among 7310 cases of Lyme disease, we classified 62% by stage. Overall, 23% were classified using both diagnoses and text, 26% were classified using diagnoses only, and 13% were classified using text only. Among the staged diagnoses (n = 4530), 30% were disseminated stage (762 arthritis, 426 neurological manifestations, 76 carditis, 95 secondary erythema migrans, and 76 other manifestations). In adjusted models, we found that persons on Medical Assistance at least 50% of time under observation, compared to never users, had a higher risk (risk ratio [95% confidence interval]) of disseminated Lyme disease (1.20 [1.05, 1.37]). Primary care contact (0.59 [0.54, 0.64]) and diagnosis in the urgent care (0.22 [0.17, 0.29]), compared to the outpatient setting, were associated with lower risk of disseminated Lyme disease. CONCLUSIONS: The associations between insurance payor, primary care status, and diagnostic setting with disseminated Lyme disease suggest that lower socioeconomic status and less health care access could be linked with disseminated stage Lyme disease. Intervening on these factors could reduce the individual and health care burden of disseminated Lyme disease. Our findings demonstrate the value of both diagnostic and narrative text data to identify Lyme disease manifestations in the EHR.

Tick CAPA propeptide cDNAs and receptor activity of endogenous tick pyrokinins and analogs: Towards discovering pyrokinin function in ticks.

Pyrokinins (PKs) are pleiotropic neuropeptides with significant roles in invertebrate physiology. Although functions of PKs are known in insects, there is a lack of knowledge of PK-encoding genes and PKs functions in ticks. Herein the first tick cDNAs of the capability (capa) gene were cloned from the southern cattle tick, Rhipicephalus microplus (Acari: Ixodidae), and the blacklegged tick, Ixodes scapularis. Each cDNA encoded one periviscerokinin and five different pyrokinins. Two PKs were identical in sequence in the two species. The three PKs unique to R. microplus (Rhimi-CAPA-PK1, -PK2, and -PK5) were tested on the recombinant R. microplus pyrokinin receptor using a calcium bioluminescence assay. The Rhimi-CAPA-PKs acted as agonists with EC50s ranging from 101-188 nM. Twenty PK analogs designed for enhanced bioavailability and biostability were tested on the receptor. Five of these were designed based on the sequences of the three unique Rhimi-CAPA-PKs. Eight PK analogs were also agonists; four of them were full agonists that exhibited comparable efficacy to the native Rhimi-CAPA-PKs, with EC50 ranging from 401 nM-1.9 µM. The structure-activity relationships (SAR) of all analogs were analyzed. Our results suggested that a positively charged, basic lysine at the variable position X of the PK active core (FXPRLamide) conferred enhanced affinity to the analogs in their interaction with the tick receptor. These analogs are promising tools to elucidate the pyrokinin function in ticks in vivo as these analogs are expected to have prolonged hemolymph residence time in comparison to the native peptides.

Acaricidal and repellent activity of the essential oils of Backhousia citriodora, Callistemon viminalis and Cinnamodendron dinisii against Rhipicephalus spp.

The ticks Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato (s.l) are of great importance to agriculture, veterinary medicine and public health. Due to a number of problems related to the use of synthetic acaricides, natural products emerge as promising substances for alternative tick control. In the present study, essential oils of Backhousia citriodora, Callistemon viminalis and Cinnamodendron dinisii were extracted by hydrodistillation, characterized by GC-MS and GC-FID and biologically evaluated for acaricidal activity against R. microplus and repellent activity against R. sanguineus s.l. Inhibition of acetylcholinesterase (AChe) by the essential oils was also evaluated. The major constituent of B. citriodora is citral (98.9 %), whereas the essential oil from C. viminalis is rich in 1.8-cineole (78.1 %), α-pinene (12.5 %) and limonene (3.36 %), and that from C. dinisii contains α-pinene (30.8 %), ß-pinene (12.5 %) and sabinene (11.3 %) as the principal constituents. The median lethal concentrations (LC 50) estimated for the essential oils on engorged R. microplus females were 3.276 µL.mL-1 for B. citriodora, 8.195 µL.mL-1 for C. dinisii and 8.936 µL.mL-1 for C. viminalis. The essential oil of B. citriodora showed the best repellent effect against unfed R. sanguineus s.l. adults, demonstrating repellent action up to 3 h after application. The essential oil of C. viminalis was able to reduce AChe activity, with an average inhibitory concentration (IC 50) of 0.33 µg mL-1. Thus, these oils can be considered as sources of bioactive compounds for tick control.