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Spirochetes were observed in the midguts of 35% of 147 motile Ixodes dammini from three locations in Lyme and East Haddam, Connecticut. Positive ticks were removed from eastern chipmunks (Tamias striatus), raccoons (Procyon lotor), white-footed mice (Peromyscus leucopus), and a red squirrel (Tamiasciurus hudsonicus). Spirochetes were isolated in fortified Kelly's medium from nine questing or partially engorged I. dammini adults and nymphs and from the bloods of a raccoon and a white-footed mouse. Connecticut isolates from ticks and mammals were serologically indistinguishable from the original Shelter Island, New York strain when cross-tested by immunofluorescence against their mouse antisera. Sera from eight patients diagnosed as having Lyme disease contained antibodies to spirochetes isolated from ticks and mammals. Our finding of serologically and morphologically indistinguishable spirochetes in a raccoon, white-footed mouse and ticks suggests that closely related serotypes are present in wild mammals commonly parasitized by I. dammini, and further supports the claim that a spirochete is involved in the etiology of Lyme disease.

Education can help improve compliance with inhaled corticosteroid therapy or correct faulty metered-dose inhaler (MDI) technique. Options for patients with poor MDI technique include use of a spacer or an alternative device, such as a nebulizer or a dry powder in-haler. If therapy is ineffective, consider alternative conditions that mimic asthma, especially vocal cord dysfunction and upper airway obstruction. Treatment of comorbid conditions, such as gastroesophageal reflux disease or rhinosinusitis, may improve control. In refractory asthma, it is crucial to identify allergic triggers and reduce exposure to allergens. If another medication needs to be added to the in-haled corticosteroid, consider a long-acting beta-agonist, leukotriene modifier, or the recombinant monoclonal anti-IgE anti-body omalizumab. [ABSTRACT FROM AUTHOR]

Soil CO 2 concentration and flux measurements are important in diverse fields, including geoscience, climate science, soil ecology, and agriculture. However, practitioners in these fields face difficulties with existing soil CO 2 gas probes, which have had problems with high costs and frequent failures when deployed. Confronted with a recent research project's need for long-term in-soil CO 2 monitoring at a large number of sites in harsh environmental conditions, we developed our own CO 2 logging system to reduce expense and avoid the expected failures of commercial instruments. Our newly developed soil probes overcome the central challenge of soil gas probes-surviving continuous exposure to soil moisture while remaining open to soil gases-via three approaches: a 3D printed housing (economical for small-scale production) following design principles that correct the usual water permeability flaw of 3D printed materials; passive moisture protection via a hydrophobic, CO 2 -permeable PTFE membrane; and active moisture protection via a low-power micro-dehumidifier. Our CO 2 instrumentation performed well and yielded a high-quality dataset that includes signals related to a prescribed fire as well as seasonal and diel cycles. We expect our technology to support underground CO 2 monitoring in fields where it is already practiced and stimulate its expansion into diverse new fields.

Background: West Nile virus (WNV), Everglades virus (EVEV), and five species of Orthobunyavirus were isolated from mosquitoes collected in the Everglades in 2016-2017. Prior studies of blood meals of mosquitoes in southern Florida have related findings to acquisition and transmission of EVEV, St. Louis encephalitis virus, and WNV, but not the Orthobunyavirus viruses associated with the subgenus Melanoconion of the genus Culex . Materials and Methods: In the present study, blood-fed mosquitoes were collected in the Everglades in 2016, 2017, 2021, and 2022, and from an industrial site in Naples, FL in 2017. Blood meals were identified to host species by PCR assays using mitochondrial cytochrome b gene. Results: Blood meals were identified from Anopheles crucians complex and 11 mosquito species captured in the Florida Everglades and from 3 species collected from an industrial site. The largest numbers of blood-fed specimens were from Culex nigripalpus , Culex erraticus , Culex cedecei , and Aedes taeniorhynchus . Cx. erraticus fed on mammals, birds, and reptiles, particularly American alligator. This mosquito species could transmit WNV to American alligator in the wild. Cx. nigripalpus acquired blood meals primarily from birds and mammals and frequently fed on medium-sized mammals and white-tailed deer. Water and wading birds were the primary avian hosts for Cx. nigripalpus and Cx. erraticus in the Everglades. Wading birds are susceptible to WNV and could serve as reservoir hosts. Cx. cedecei fed on five species of rodents, particularly black and hispid cotton rats. EVEV and three different species of Orthobunyavirus have been isolated from the hispid cotton rat and Cx. cedecei in the Everglades. Cx. cedecei is likely acquiring and transmitting these viruses among hispid cotton rats and other rodents. The marsh rabbit was a frequent host for An. crucians complex. An. crucians complex, and other species could acquire Tensaw virus from rabbits. Conclusions: Our study contributes to a better understanding of the host and viral associations of mosquito species in southwestern Florida.

Brazoran virus was first isolated from Culex mosquitoes in Texas in 2012, yet little is known about this virus. We report the isolation of this virus from Culex erraticus from southern Florida during 2016. The Florida strain had a nucleotide identity of 96.3% (S segment), 99.1% (M segment), and 95.8% (L segment) to the Texas isolate. Culex quinquefasciatus and Aedes aegypti colonies were subsequently fed virus blood meals to determine their vector competence for Brazoran virus. Culex quinquefasciatus was susceptible to midgut infection, but few mosquitoes developed disseminated infections. Aedes aegypti supported disseminated infection, but virus transmission could not be demonstrated. Suckling mice became infected by intradermal inoculation without visible disease signs. The virus was detected in multiple mouse tissues but rarely infected the brain. This study documents the first isolation of Brazoran virus outside of Texas. Although this virus infected Ae. aegypti and Cx. quinquefasciatus in laboratory trials, their vector competence could not be demonstrated, suggesting they are unlikely vectors of Brazoran virus.

Thirty-seven species and subspecies of mosquitoes were identified from 3,580,610 specimens collected in eastern (Cass, Nelson, and Richland counties) and western (Williams County) North Dakota in 2003-2006. Four species were new state records (Aedes schizopinax, Psorophora ciliata, Ps. ferox, and Ps. horrida). Aedes vexans was dominant (82.9%). Other relatively abundant species were Ae. trivittatus (7.7%), Ae. melanimon (2.7%), Culex tarsalis (2.6%), Ae. dorsalis (1.6%), Ae. sticticus (1.0), and Culiseta inornata (0.9%). The seasonality of the species is presented., (Copyright © 2023 by The American Mosquito Control Association, Inc.)

Children's ability to recognize object shape is foundational for successful early word learning. However, the prototypical shape of objects may not be easily accessible-take margarita glasses, for instance. The current study examined 304 U.S. children 17- to 42-month-old (152 females) from 2017 to 2020, asking how shape, age, and vocabulary abilities predict recognition of everyday objects. Children's ability to recognize objects increases with age and vocabulary, replicating prior work. Moreover, performance was partially moderated by object's typicality and shape features, and children's own attention to shape (shape bias) may mediate the effect, especially with prototypically shaped objects. The current study highlights how both child-specific variables and context features interact to shape language abilities, underscoring the emergent and multi-causal nature of word learning., (© 2022 The Authors. Child Development © 2022 Society for Research in Child Development.)

Infrasound may be used to detect the approach of hazardous volcanic mudflows, known as lahars, tens of minutes before their flow fronts arrive. We have analyzed signals from more than 20 secondary lahars caused by precipitation events at Fuego Volcano during Guatemala's rainy season in May through October of 2022. We are able to quantify the capabilities of infrasound monitoring through comparison with seismic data, time lapse camera imagery, and high-resolution video of a well-recorded event on August 17. We determine that infrasound sensors, deployed adjacent to the lahar path and in small-aperture (10 s of meters) arrays, are particularly sensitive to remote detection of lahars, including small-sized events, at distances of at least 5 km. At Fuego Volcano these detections could be used to provide timely alerts of up to 30 min before lahars arrive at a downstream monitoring site, such as in the frequently impacted Ceniza drainage. We propose that continuous infrasound monitoring, from locations adjacent to a drainage, may complement seismic monitoring and serve as a valuable tool to help identify approaching hazards. On the other hand, infrasound arrays located a kilometer or more from the lahar path can be effectively used to track a lahar's progression., (© 2023. The Author(s).)

Powassan virus is an emerging tick-borne virus of concern for public health, but very little is known about its transmission patterns and ecology. Here, we expanded the genomic dataset by sequencing 279 Powassan viruses isolated from Ixodes scapularis ticks from the northeastern United States. Our phylogeographic reconstructions revealed that Powassan virus lineage II was likely introduced or emerged from a relict population in the Northeast between 1940 and 1975. Sequences strongly clustered by sampling location, suggesting a highly focal geographical distribution. Our analyses further indicated that Powassan virus lineage II emerged in the northeastern United States mostly following a south-to-north pattern, with a weighted lineage dispersal velocity of ~3 km/y. Since the emergence in the Northeast, we found an overall increase in the effective population size of Powassan virus lineage II, but with growth stagnating during recent years. The cascading effect of population expansion of white-tailed deer and I. scapularis populations likely facilitated the emergence of Powassan virus in the northeastern United States.

Ixodes scapula ris is a medically important tick that transmits several microbes to humans, including rickettsial pathogen Anaplasma phagocytophilum . In nature, these ticks encounter several abiotic factors including changes in temperature, humidity, and light. Many organisms use endogenously generated circadian pathways to encounter abiotic factors. In this study, we provide evidence for the first time to show that A. phagocytophilum modulates the arthropod circadian gene for its transmission to the vertebrate host. We noted a circadian oscillation in the expression of arthropod clock , bmal1 , period and timeless genes when ticks or tick cells were exposed to alternate 12 h light: 12 h dark conditions. Moreover, A. phagocytophilum significantly modulates the oscillation pattern of expression of these genes. In addition, increased levels of clock and bmal1 and decreased expression of Toll and JAK/STAT pathway immune genes such as pelle and jak, respectively, were noted during A. phagocytophilum transmission from ticks to the vertebrate host. RNAi-mediated knockdown of clock gene expression in ticks resulted in the reduced expression of jak and pelle that increased bacterial transmission from ticks to the murine host. Furthermore, clock -deficient ticks fed late and had less engorgement weights. These results indicate an important role for circadian modulation of tick gene expression that is critical for arthropod blood feeding and transmission of pathogens from vector to the vertebrate host.

Mosquitoes were collected for 12 consecutive months beginning June 2016, from 11 locations in the Florida Everglades, Collier County, and tested for viruses by isolation in Vero cells and subsequent identification. One species complex and 31 species of mosquitoes were identified from 668,809 specimens. Ochlerotatus taeniorhynchus comprised 72.2% of the collection. Other notable species were Anopheles crucians complex, Culex nigripalpus, Cx. erraticus, and Cx. cedecei. Seven species of virus were identified from 110 isolations: Everglades, Gumbo Limbo, Mahogany Hammock, Pahayokee, Shark River, Tensaw, and West Nile viruses. Everglades, West Nile, Tensaw, and Mahogany Hammock viruses were most frequently isolated. Largest numbers of viruses were identified from Cx. cedecei, Cx. nigripalpus, and An. crucians complex. Five species of virus were isolated from Cx. cedecei. Viruses were isolated from mangrove, cypress swamp, hardwood hammock, and sawgrass habitats. West Nile virus was isolated August through October when Cx. nigripalpus was most abundant. Everglades virus was the most frequently isolated virus from nine species of mosquitoes collected from June through August. Tensaw virus was isolated primarily from Anopheles species. Isolations were made in July, August, January, February, and April, suggesting that this virus may be present in host-seeking mosquitoes throughout the year. Mahogany Hammock, Shark River, Gumbo Limbo, and Pahayokee viruses were isolated primarily from Cx. cedecei from June through December. Shotgun metagenomic sequencing was used to document that seven pools of Cx. cedecei were infected with two arboviruses. As communities expand into the Everglades, more humans will become exposed to arboviruses.

Reactive oxygen species (ROS) that are induced upon pathogen infection plays an important role in host defence. The rickettsial pathogen Anaplasma phagocytophilum, which is primarily transmitted by Ixodes scapularis ticks in the United States, has evolved many strategies to escape ROS and survive in mammalian cells. However, little is known on the role of ROS in A. phagocytophilum infection in ticks. Our results show that A. phagocytophilum and hemin induce activation of l-tryptophan pathway in tick cells. Xanthurenic acid (XA), a tryptophan metabolite, supports A. phagocytophilum growth in tick cells through inhibition of tryptophan dioxygenase (TDO) activity leading to reduced l-kynurenine levels that subsequently affects build-up of ROS. However, hemin supports A. phagocytophilum growth in tick cells by inducing TDO activity leading to increased l-kynurenine levels and ROS production. Our data reveal that XA and kynurenic acid (KA) chelate hemin. Furthermore, treatment of tick cells with 3-hydroxyl l-kynurenine limits A. phagocytophilum growth in tick cells. RNAi-mediated knockdown of kynurenine aminotransferase expression results in increased ROS production and reduced A. phagocytophilum burden in tick cells. Collectively, these results suggest that l-tryptophan pathway metabolites influence A. phagocytophilum survival by affecting build up of ROS levels in tick cells., (© 2020 John Wiley & Sons Ltd.)

West Nile virus (family Flaviviridae, genus Flavivirus) first caused human and veterinary disease, and was isolated from Culex pipiens pipiens L. and Aedes vexans (Meigen) (Diptera: Culicidae) in the United States in 1999. We report that a Connecticut strain of Ae. vexans was competent to transmit West Nile virus both horizontally to suckling mice and vertically to its progeny in the laboratory. Horizontal transmission was first observed on day 6 post-exposure (pe). Daily horizontal transmission rates generally increased with the day post-virus exposure with highest rates of 67-100% recorded on days 28-30 pe. One female vertically transmitted West Nile virus on day 21 pe, but only after it had taken its third bloodmeal. Horizontal and vertical transmission may contribute to West Nile virus infection rates in Ae. vexans in summer, and vertical transmission provides a means of survival of West Nile virus during winter., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Purpose: Computed tomographic pulmonary angiography (CTPA) is the test of choice for patients with acute chest pain and suspected pulmonary embolism (PE). This examination is excellent for the diagnosis of PE and can also often identify alternative diagnoses. The early phase of contrast, however, may not allow for optimal evaluation of lymph nodes, serosal surfaces, and solid organs, leading to the nonvisualization of important findings and the potential for missed diagnoses. The purpose of this study was to determine the frequency of relevant findings only identified on standard portal venous phase CT compared with CTPA., Materials and Methods: The reports for all patients in the previous 10 years who underwent both standard CT and CTPA within 7 days, for a total of 675 pairs of scans, were tabulated according to the presence of PE, serosal abnormalities, solid organ abnormalities, and lymphadenopathy. All findings were categorized as present on both scans, standard CT only, or CTPA only. The scans were manually evaluated to exclude findings that were new or resolved on the second study or outside the field of view on one of the studies., Results: Significantly more PEs were identified only on CTPA examinations. However, significantly more pleural, peritoneal, and solid organ abnormalities, and abnormal mediastinal and abdominal lymph nodes were identified on standard CT only. There was no significant difference in the identification of pericardial abnormalities or abnormal hilar lymph nodes between the two scans., Conclusions: Many serosal abnormalities, solid organ abnormalities, and lymphadenopathy were only reported on standard portal venous phase CT compared with CTPA.

The microRNAs (miRNAs) are important regulators of gene expression. In this study, we provide evidence for the first time to show that rickettsial pathogen Anaplasma phagocytophilum infection results in the down-regulation of tick microRNA-133 (miR-133), to induce Ixodes scapularis organic anion transporting polypeptide (isoatp4056) gene expression critical for this bacterial survival in the vector and for its transmission to the vertebrate host. Transfection studies with recombinant constructs containing transcriptional fusions confirmed binding of miR-133 to isoatp4056 mRNA. Treatment with miR-133 inhibitor resulted in increased bacterial burden and isoatp4056 expression in ticks and tick cells. In contrast, treatment with miR-133 mimic or pre-mir-133 resulted in dramatic reduction in isoatp4056 expression and bacterial burden in ticks and tick cells. Moreover, treatment of ticks with pre-mir-133 affected vector-mediated A. phagocytophilum infection of murine host. These results provide novel insights to understand impact of modulation of tick miRNAs on pathogen colonization in the vector and their transmission to infect the vertebrate host., Competing Interests: The authors have declared that no competing interests exist.

When bad things happen people often seek out close others for support to help regulate their negative emotions. The degree to which support providers are responsive to the specific needs of support seekers is associated with many outcomes, including how effective that support is in regulating emotion. The ability of support providers to accurately assess the emotions experienced by support seekers seems crucial, yet few studies have examined the role this type of accuracy plays in support provision. We predicted that individuals who accurately assessed the emotions being experienced by a support seeker would provide more responsive support. Further, we predicted that individual differences in emotion differentiation (perceiving differences between similar emotions), range (experiencing a range of emotions), and clarity (understanding the cause and effect of one's emotions) would facilitate emotional accuracy and, in turn, responsiveness. Participants read scenarios depicting their romantic partners seeking support to help regulate different negative emotions; they then wrote supportive messages and indicated which emotions they thought their partners would be experiencing. Individual differences in emotional range and clarity (but not differentiation) predicted how accurate participants were in gaging the emotions depicted in the scenarios. In turn, accuracy predicted how responsive their messages were, as rated by independent coders. These results suggest that accuracy in perceiving a partner's emotions is crucial for providing responsive support and individual differences in one's own emotional experiences are associated both accuracy and responsiveness. Our findings have implications for research on interpersonal emotion regulation, close relationships, and social support. (PsycInfo Database Record (c) 2020 APA, all rights reserved).

Lyme disease is the most prevalent vector-borne disease in the United States. Ixodes scapularis, commonly referred to as the blacklegged tick, is the primary vector of Lyme disease spirochetes, Borrelia burgdorferi sensu lato (s.l.), in the eastern United States. Connecticut has pervasive populations of I. scapularis and remains a hotspot for Lyme disease. A primary aim of this study was to determine if passively collected data on human-biting I. scapularis ticks in Connecticut could serve as a useful proxy for Lyme disease incidence based on the cases reported by the Connecticut Department of Public Health (CDPH). Data for human-biting I. scapularis ticks submitted to the Tick Testing Laboratory at the Connecticut Agricultural Experiment Station (CAES-TTL), and tested for infection with B. burgdorferi s.l., were used to estimate the rate of submitted nymphs, nymphal infection prevalence, and the rate of submitted infected nymphs. We assessed spatiotemporal patterns in tick-based measures and Lyme disease incidence with generalized linear and spatial models. In conjunction with land cover and household income data, we used generalized linear mixed effects models to examine the association between tick-based risk estimates and Lyme disease incidence. Between 2007 and 2017, the CAES-TTL received 26,116 I. scapularis tick submissions and the CDPH reported 23,423 Lyme disease cases. The rate of submitted nymphs, nymphal infection prevalence, the rate of submitted infected nymphs, and Lyme disease incidence all decreased over time during this eleven-year period. The rate of submitted nymphs, the rate of submitted infected nymphs, and Lyme disease incidence were spatially correlated, but nymphal infection prevalence was not. Using a mixed modeling approach to predict Lyme disease incidence and account for spatiotemporal structuring of the data, we found the best fitting tested model included a strong, positive association with the rate of submitted infected nymphs and a negative association with the percent of developed land for each county. We show that within counties, submissions of B. burgdorferi s.l. infected nymphs were strongly and positively associated with inter-annual variation in reported Lyme disease cases. Tick-based passive surveillance programs may be useful in providing independent measures of entomological risk, particularly in settings where Lyme disease case reporting practices change substantially over time., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

Drought conditions affect ozone air quality, potentially altering multiple terms in the O 3 mass balance equation. Here, we present a multiyear observational analysis using data collected before, during, and after the record-breaking California drought (2011-2015) at the O 3 -polluted locations of Fresno and Bakersfield near the Sierra Nevada foothills. We separately assess drought influences on O 3 chemical production ( PO 3 ) from O 3 concentration. We show that isoprene concentrations, which are a source of O 3 -forming organic reactivity, were relatively insensitive to early drought conditions but decreased by more than 50% during the most severe drought years (2014-2015), with recovery a function of location. We find drought-isoprene effects are temperature-dependent, even after accounting for changes in leaf area, consistent with laboratory studies but not previously observed at landscape scales with atmospheric observations. Drought-driven decreases in organic reactivity are contemporaneous with a change in dominant oxidation mechanism, with PO 3 becoming more NO x -suppressed, leading to a decrease in PO 3 of ∼20%. We infer reductions in atmospheric O 3 loss of ∼15% during the most severe drought period, consistent with past observations of decreases in O 3 uptake by plants. We consider drought-related trends in O 3 variability on synoptic time scales by analyzing statistics of multiday high-O 3 events. We discuss implications for regulating O 3 air pollution in California and other locations under more prevalent drought conditions.

Background: Ixodes scapularis organic anion transporting polypeptides (OATPs) play important roles in tick-rickettsial pathogen interactions. In this report, we characterized the role of these conserved molecules in ticks infected with either Lyme disease agent Borrelia burgdorferi or tick-borne Langat virus (LGTV), a pathogen closely related to tick-borne encephalitis virus (TBEV)., Results: Quantitative real-time polymerase chain reaction analysis revealed no significant changes in oatps gene expression upon infection with B. burgdorferi in unfed ticks. Synchronous infection of unfed nymphal ticks with LGTV in vitro revealed no significant changes in oatps gene expression. However, expression of specific oatps was significantly downregulated upon LGTV infection of tick cells in vitro. Treatment of tick cells with OATP inhibitor significantly reduced LGTV loads, kynurenine amino transferase (kat), a gene involved in the production of tryptophan metabolite xanthurenic acid (XA), levels and expression of several oatps in tick cells. Furthermore, bioinformatics characterization of OATPs from some of the medically important vectors including ticks, mosquitoes and lice revealed the presence of several glycosylation, phosphorylation and myristoylation sites., Conclusions: This study provides additional evidence on the role of arthropod OATPs in vector-intracellular pathogen interactions.

Lyme disease, caused by the spirochetal bacterium, Borrelia burgdorferi sensu lato (Bbsl), is typically transmitted by hard-bodied ticks (Acari: Ixodidae). Whenever this tick-borne zoonosis is mentioned in medical clinics and emergency rooms, it sparks a firestorm of controversy. Denial often sets in, and healthcare practitioners dismiss the fact that this pathogenic spirochetosis is present in their area. For distribution of Bbsl across Canada, we conducted a 4-year, tick⁻host study (2013⁻2016), and collected ticks from avian and mammalian hosts from Atlantic Canada to the West Coast. Overall, 1265 ticks representing 27 tick species belonging to four genera were collected. Of the 18 tick species tested, 15 species (83%) were positive for Bbsl and, of these infected ticks, 6 species bite humans. Overall, 13 of 18 tick species tested are human-biting ticks. Our data suggest that a 6-tick, enzootic maintenance cycle of Bbsl is present in southwestern B.C., and five of these tick species bite humans. Biogeographically, the groundhog tick, Ixodes cookei , has extended its home range from central and eastern Canada to southwestern British Columbia (B.C.). We posit that the Fox Sparrow, Passerella iliaca , is a reservoir-competent host for Bbsl. The Bay-breasted Warbler, Setophaga castanea , and the Tennessee Warbler, Vermivora peregrina , are new host records for the blacklegged tick, Ixodes scapularis . We provide the first report of a Bbsl-positive Amblyomma longirostre larva parasitizing a bird; this bird parasitism suggests that a Willow Flycatcher is a competent reservoir of Bbsl. Our findings show that Bbsl is present in all provinces, and that multiple tick species are implicated in the enzootic maintenance cycle of this pathogen. Ultimately, Bbsl poses a serious public health contagion Canada-wide.

A simple, effective light scattering prototype sensitive to both polarized and depolarized scattering was constructed, and its performance was tested on a variety of pure liquids and optically isotropic and anisotropic polymer solutions and colloidal suspensions. The results, performance, and means to further improvement are reported here. Because of its simplicity and low cost, many identical units can be produced to construct a simultaneous multiple sample light scattering platform that can be used to monitor polymer and colloid solution stability, phase changes, aggregation, degradation, etc. Measurable depolarization was found for a variety of organic liquids and suspensions of both polyfluoroethylene and latex spheres. No detectable depolarization was found for various polymers.

Competing Interests: Competing interests: None declared.

Ixodes scapularis ticks transmit several pathogens to humans including rickettsial bacterium, Anaplasma phagocytophilum. Here, we report that A. phagocytophilum uses tick transcriptional activator protein-1 (AP-1) as a molecular switch in the regulation of arthropod antifreeze gene, iafgp. RNAi-mediated silencing of ap-1 expression significantly affected iafgp gene expression and A. phagocytophilum burden in ticks upon acquisition from the murine host. Gel shift assays provide evidence that both the bacterium and AP-1 influences iafgp promoter and expression. The luciferase assays revealed that a region of approximately 700 bp upstream of the antifreeze gene is sufficient for AP-1 binding to promote iafgp gene expression. Furthermore, survival assays revealed that AP-1-deficient ticks were more susceptible to cold in comparison to the mock controls. In addition, this study also indicates arthropod AP-1 as a global regulator for some of the tick genes critical for A. phagocytophilum survival in the vector. In summary, our study defines a novel mode of arthropod signaling for the survival of both rickettsial pathogen and its medically important vector in the cold.

We report the importation into Connecticut, U.S.A., of an exotic tick, Hyalomma truncatum (Koch) (Acari: Ixodidae), on a human with recent travel history to Africa. The tick was identified using key morphological characters and through DNA sequencing. This case report highlights continuing risk associated with the importation of exotic tick vectors of medical and veterinary significance on international travelers returning to the United States from abroad.

Bed bugs, Cimex lectularius L. (Hemiptera: Cimicidae), have become a major health nuisance in the past 20 ysin cities and elsewhere throughout many areas of the world. Few studies have reported on repellent compounds that could reduce their transport in luggage. We evaluated the repellency of six naturally occurring or related compounds used in flavor/fragrance applications or structurally related compounds, para-menthane-3,8-diol, and N,N-diethyl-meta-toluamide (DEET) to bed bugs in a 183 × 183-cm arena . Repellency was assessed using soft-sided polyester lunch bags serving as surrogates of luggage and barrier cloth towels upon which rested untreated lunch bags. We report for the first time repellency of delta dodecalactone, 2-(3, 7-dimethyl-2, 6-nonadien-1-yl)-cyclopentanone (a.k.a. 'methyl apritone'), gamma dodecalactone, and para-menthane-3,8-diol to bed bugs. Propyl dihydrojasmonate, 3-methyl-5-hexyl-2-cyclohexenone, gamma methyl tridecalactone, and DEET are also documented to be repellent to bed bugs. These compounds provided relatively long-term protection. Propyl dihydrojasmonate prevented bed bugs from seeking refuge in treated lunch bags 27 d after treatment, and when applied to cloth towels repelled harborage-seeking bed bugs for 146 d. Methyl apritone blended with 3-methyl-5-hexyl-2-cyclohexenone and delta dodecalactone as an individual compound applied to cloth towels repelled bed bugs for 190 and 276 d, respectively. The above-mentioned compounds, either individually or as blends, may reduce risk of bed bugs seeking harborage in treated suitcases or towels upon which untreated luggage is placed.

Seasonal abundance of mosquitoes, their viruses, and blood-feeding habits were determined at an open-faced quarry in North Branford, CT, in 2010 and 2011. This unique habitat had not previously been sampled for mosquitoes and mosquito-borne viruses. Thirty species of mosquitoes were identified from 41,719 specimens collected. Coquillettidia perturbans , Aedes trivittatus , and Ae. vexans were the most abundant species and represented 34.5%, 17.7%, and 14.8% of the totals, respectively. Jamestown Canyon virus was isolated from 6 species of mosquitoes collected from mid-June through July: Cq. perturbans (3 pools), Ae. cantator (3), Ae. trivittatus (2), Ae. aurifer (1), Ae. excrucians (1), and Culex pipiens (1). West Nile virus was cultured from 8 pools of Cx. pipiens and from 1 pool of Culiseta melanura collected from mid-August through late September. Cache Valley virus was isolated from 4 species of mosquitoes in 3 genera from about mid-August through late September 2011: Cq. perturbans (5 pools), Ae. trivittatus (2), Anopheles punctipennis (1), and An. quadrimaculatus (1). Nine different mammalian hosts were identified as sources of blood for 13 species of mosquitoes. White-tailed deer, Odocoileus virginianus , were the most common mammalian hosts (90.8%), followed by raccoon, Procyon lotor (3.1%), coyote, Canis latrans (2.4%), and human, Homo sapiens (1.2%). Exclusive mammalian blood-feeding mosquitoes included: Ae. canadensis , Ae. cantator , Ae. excrucians , Ae. japonicus , Ae. vexans , An. punctipennis , and Cx. salinarius . Fourteen species of birds, mostly Passeriformes, were identified as sources of blood from 6 mosquito species. Five species that fed on mammals ( Ae. thibaulti , Ae. trivittatus , Ae. cinereus , Cq. perturbans , and Cx. pipiens ) also fed on birds.

Trace elements such as copper and cobalt have been associated with virus-host interactions. However, studies to show the effect of conjugation of copper(II) or cobalt(III) metal centers to thiosemicarbazone ligand(s) derived from either food additives or mosquito repellent such as 2-acetylethiazole or citral, respectively, on Zika virus (ZIKV) or dengue virus (serotype 2; DENV2) infections have not been explored. In this study, we show that four compounds comprising of thiosemicarbazone ligand derived from 2-acetylethiazole viz., (E)-N-ethyl-2-[1-(thiazol-2-yl)ethylidene]hydrazinecarbothioamide (acetylethTSC) (compound 1), a copper(II) complex with acetylethTSC as a ligand (compound 2), a thiosemicarbazone ligand-derived from citral (compound 3) and a cobalt(III) complex with a citral-thiosemicarbazone ligand (compound 4) increased DENV2 and ZIKV replication in both mosquito C6/36 cells and human keratinocytes (HaCaT cells). Treatment of both cell lines with compounds 2 or 4 showed increased dengue viral titers at all three tested doses. Enhanced dengue viral plaque formation was also noted at the tested dose of 100μM, suggesting higher production of infectious viral particles. Treatment with the compounds 2 or 4 enhanced ZIKV and DENV2 RNA levels in HeLa cell line and primary cultures of mouse bone marrow derived dendritic cells. Also, pre- or post treatments with conjugated compounds 2 or 4 showed higher loads of ZIKV or DENV2 envelope (E) protein in HaCaT cells. No changes in loads of E-protein were found in ZIKV-infected C6/36 cells, when compounds were treated after infection. In addition, we tested bis(1,10-phenanthroline)copper(II) chloride ([Cu(phen) 2 ]Cl 2 , (compound 5) and tris(1,10-phenanthroline)cobalt(III) chloride ([Co(phen) 3 ]Cl 3 , (compound 6) that also showed enhanced DENV2 loads. Also, we found that copper(II) chloride dehydrate (CuCl 2 ·2H 2 O) or cobalt(II) chloride hexahydrate (CoCl 2 ·6H 2 O) alone had no effects as "free" cations. Taken together, these findings suggest that use of Cu(II) or Co(III) conjugation to organic compounds, in insect repellents and/or food additives could enhance DENV2/ZIKV loads in human cells and perhaps induce pathogenesis in infected individuals or individuals pre-exposed to such conjugated complexes., Importance: Mosquito-borne diseases are of great concern to the mankind. Use of chemicals/repellents against mosquito bites and transmission of microbes has been the topic of interest for many years. Here, we show that thiosemicarbazone ligand(s) derived from 2-acetylethiazole or citral or 1,10-phenanthroline upon conjugation with copper(II) or cobalt(III) metal centers enhances dengue virus (serotype 2; DENV2) and/or Zika virus (ZIKV) infections in mosquito, mouse and human cells. Enhanced ZIKV/DENV2 capsid mRNA or envelope protein loads were evident in mosquito cells and human keratinocytes, when treated with compounds before/after infections. Also, treatment with copper(II) or cobalt(III) conjugated compounds increased viral titers and number of plaque formations. These studies suggest that conjugation of compounds in repellents/essential oils/natural products/food additives with copper(II) or cobalt(III) metal centers may not be safe, especially in tropical and subtropical places, where several dengue infection cases and deaths are reported annually or in places with increased ZIKV caused microcephaly., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The black-legged tick Ixodes scapularis transmits the human anaplasmosis agent, Anaplasma phagocytophilum. In this study, we show that A. phagocytophilum specifically up-regulates I. scapularis organic anion transporting polypeptide, isoatp4056 and kynurenine amino transferase (kat), a gene involved in the production of tryptophan metabolite xanthurenic acid (XA), for its survival in ticks. RNAi analysis revealed that knockdown of isoatp4056 expression had no effect on A. phagocytophilum acquisition from the murine host but affected the bacterial survival in tick cells. Knockdown of the expression of kat mRNA alone or in combination with isoatp4056 mRNA significantly affected A. phagocytophilum survival and isoatp4056 expression in tick cells. Exogenous addition of XA induces isoatp4056 expression and A. phagocytophilum burden in both tick salivary glands and tick cells. Electrophoretic mobility shift assays provide further evidence that A. phagocytophilum and XA influences isoatp4056 expression. Collectively, this study provides important novel information in understanding the interplay between molecular pathways manipulated by a rickettsial pathogen to survive in its arthropod vector.

Research on automatic processes in fear has emphasized the provocation of fear responses rather than their attenuation. We have previously shown that the repeated presentation of feared images without conscious awareness via backward masking reduces avoidance of a live tarantula in spider-phobic participants. Herein we investigated the neural basis for these adaptive effects of masked exposure. 21 spider-phobic and 21 control participants, identified by a psychiatric interview, fear questionnaire, and approaching a live tarantula, viewed stimuli in each of three conditions: (1) very brief exposure (VBE) to masked images of spiders, severely limited awareness; (2) clearly visible exposure (CVE) to spiders, full awareness; and (3) masked images of flowers (control), severely limited awareness. Only VBE to masked spiders generated neural activity more strongly in phobic than in control participants, within subcortical fear, attention, higher-order language, and vision systems. Moreover, VBE activated regions that support fear processing in phobic participants without causing them to experience fear consciously. Counter-intuitively, CVE to the same spiders generated stronger neural activity in control rather than phobic participants within these and other systems. CVE deactivated regions supporting fear regulation and caused phobic participants to experience fear. CVE-induced activations also correlated with measures of explicit fear ratings, whereas VBE-induced activations correlated with measures of implicit fear (color-naming interference of spider words). These multiple dissociations between the effects of VBE and CVE to spiders suggest that limiting awareness of exposure to phobic stimuli through visual masking paradoxically facilitates their processing, while simultaneously minimizing the experience of fear. Hum Brain Mapp 38:2466-2481, 2017. © 2017 Wiley Periodicals, Inc., (© 2017 Wiley Periodicals, Inc.)

Ticks secrete several anti-hemostatic factors in their saliva to suppress the host innate and acquired immune defenses against infestations. Using Ixodes scapularis ticks and age-matched mice purchased from two independent commercial vendors with two different immune backgrounds as a model, we show that ticks fed on immunodeficient animals demonstrate decreased fibrinogenolytic activity in comparison to ticks fed on immunocompetent animals. Reduced levels of D-dimer (fibrin degradation product) were evident in ticks fed on immunodeficient animals in comparison to ticks fed on immunocompetent animals. Increased engorgement weights were noted for ticks fed on immunodeficient animals in comparison to ticks fed on immunocompetent animals. Furthermore, the LC-MS/MS and quantitative real-time-PCR analysis followed by inhibitor and antibody-blocking assays revealed that the arthropod HSP70-like molecule contributes to differential fibrinogenolysis during tick feeding. Collectively, these results not only indicate that ticks elicit variable fibrinogenolysis upon feeding on hosts with different immune backgrounds but also provide insights for the novel role of arthropod HSP70-like molecule in fibrinogenolysis during blood feeding.

Bed bugs, Cimex lectularius L., feed on humans, have increased exponentially in the past two decades in many major cities throughout the world, have caused intense infestations, and have become a significant health concern. Improved traps are needed to detect early infestations, to assess control programs, and for control of bed bugs. Carbon dioxide released alone or simultaneously with other attractants into three types of traps at the relatively low rate of 1 ml/min caught significantly more bed bugs than untreated controls in a 183- by 183-cm arena. This finding may enable CO2 to be used more economically in traps. Three percent ammonium bicarbonate released at a rate of ≤0.03 ml/h also caught significantly more bed bugs than untreated controls. A blend of (E)-2-hexenal and (E)-2-octenal at concentrations of 0.025% or 0.1% each and released at 0.02 ml/h attracted significantly more bed bugs than untreated controls. These findings ought to improve detection of bed bugs., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

We document the presence of blacklegged ticks, Ixodes scapularis , in the Grand River valley, Centre Wellington, Ontario. Overall, 15 (36%) of 42 I. scapularis adults collected from 41 mammalian hosts (dogs, cats, humans) were positive for the Lyme disease bacterium, Borrelia burgdorferi sensu lato (s.l.). Using real-time PCR testing and DNA sequencing of the flagellin ( fla ) gene, we determined that Borrelia amplicons extracted from I. scapularis adults belonged to B. burgdorferi sensu stricto (s.s.), which is pathogenic to humans and certain domestic animals. Based on the distribution of I. scapularis adults within the river basin, it appears likely that migratory birds provide an annual influx of I. scapularis immatures during northward spring migration. Health-care providers need to be aware that local residents can present with Lyme disease symptoms anytime during the year., Competing Interests: Competing Interests: The authors confirm that they have no conflicts of interest that could bias any aspect of the present paper. They have no financial interests in, relationships with, or have received no funding from any clinical laboratories or test kit companies or reagent manufactures mentioned in the paper.

Lyme disease is a serious health problem, with many patients requiring in-depth clinical assessment and extended treatment. In the present study, we provide the first records of the western blacklegged tick, Ixodes pacificus , and Ixodes spinipalpis parasitizing eastern cottontails, Sylvilagus floridanus . We also documented a triple co-infestation of 3 tick species (Ixodes angustus, I. pacificus , I. spinipalpis) feeding on an eastern cottontail. Notably, we discovered a unique member of the Lyme disease bacterium, Borrelia burgdorferi sensu lato (s.l.) in Canada. Ixodes spinipalpis ticks, which were collected from an eastern cottontail on Vancouver Island, British Columbia (BC), were positive for B. burgdorferi s.l. With the use of polymerase chain reaction amplification on the tick extracts and DNA sequencing on the borrelial amplicons, we detected Borrelia genomospecies 2, a novel subgroup of the B. burgdorferi s.l. complex. Based on 416 nucleotides of the flagellin B (flaB) gene, our amplicons are identical to the Borrelia genomospecies 2 type strain CA28. Borrelia genomospecies 2 is closely related genetically to other B. burgdorferi s.l. genospecies, namely Borrelia americana, Borrelia andersonii, and B. burgdorferi sensu stricto (s.s.) that cause Lyme disease. Like some other borrelial strains, Borrelia genomospecies 2 can be missed by current Lyme disease serology. Health-care providers must be aware that Borrelia genomospecies 2 is present in I. pacificus and I. spinipalpis ticks in far-western North America, and patients with clinical symptoms of Lyme disease need to be assessed for potential infection with this pathogen.

We document an established population of blacklegged ticks, Ixodes scapularis , on Corkscrew Island, Kenora District, Ontario, Canada. Primers of the outer surface protein A ( OspA ) gene, the flagellin ( fla ) gene, and the flagellin B ( flaB ) gene were used in the PCR assays to detect Borrelia burgdorferi sensu lato (s.l.), the Lyme disease bacterium. In all, 60 (73%) of 82 adult I. scapularis , were infected with B. burgdorferi s.l. As well, 6 (43%) of 14 unfed I. scapularis nymphs were positive for B. burgdorferi s.l. An I. scapularis larva was also collected from a deer mouse, and several unfed larvae were gathered by flagging leaf litter. Based on DNA sequencing of randomly selected Borrelia amplicons from six nymphal and adult I. scapularis ticks, primers for the flagellin ( fla ) and flagellin B ( flaB ) genes reveal the presence of B. burgdorferi sensu stricto (s.s.), a genospecies pathogenic to humans and certain domestic animals. We collected all 3 host-feeding life stages of I. scapularis in a single year, and report the northernmost established population of I. scapularis in Ontario. Corkscrew Island is hyperendemic for Lyme disease and has the highest prevalence of B. burgdorferi s.l. for any established population in Canada. Because of this very high infection prevalence, this population of I. scapularis has likely been established for decades. Of epidemiological significance, cottage owners, island visitors, outdoors enthusiasts, and medical professionals must be vigilant that B. burgdorferi s.l.-infected I. scapularis on Corkscrew Island pose a serious public health risk., Competing Interests: The authors have declared that no competing interest exists.

Arthropod-borne RNA viruses exist within hosts as heterogeneous populations of viral variants and, as a result, possess great genetic plasticity. Understanding the micro-evolutionary forces shaping these viruses can provide insights into how they emerge, adapt, and persist in new and changing ecological niches. While considerable attention has been directed toward studying the population dynamics of mosquito-borne viruses, little is known about tick-borne virus populations. Therefore, using a mouse and Ixodes scapularis tick transmission model, we examined Powassan virus (POWV; Flaviviridae, Flavivirus ) populations in and between both the vertebrate host and arthropod vector. We found that genetic bottlenecks, RNAi-mediated diversification, and selective constraints collectively influence POWV evolution. Together, our data provide a mechanistic explanation for the slow, long-term evolutionary trends of POWV, and suggest that all arthropod-borne viruses encounter similar selective pressures at the molecular level (i.e. RNAi), yet evolve much differently due to their unique rates and modes of transmission.

Lyme disease has emerged as a major health concern in Canada, where the etiological agent, Borrelia burgdorferi sensu lato (s.l.), a spirochetal bacterium, is typically spread by the bite of certain ticks. This study explores the presence of B. burgdorferi s.l. in blacklegged ticks, Ixodes scapularis, collected at Dundas, Ontario (a locality within the region of Hamilton-Wentworth). Using passive surveillance, veterinarians and pet groomers were asked to collect blacklegged ticks from dogs and cats with no history of travel. Additionally, I. scapularis specimens were submitted from local residents and collected by flagging. Overall, 12 (41%) of 29 blacklegged ticks were infected with B. burgdorferi s.l. Using polymerase chain reaction (PCR) and DNA sequencing, two borrelial amplicons were characterized as B. burgdorferi sensu stricto (s.s.), a genospecies pathogenic to humans and certain domestic animals. Notably, three different vertebrate hosts each had two engorged I. scapularis females removed on the same day and, likewise, one cat had three repeat occurrences of this tick species. These multiple infestations suggest that a population of I. scapularis may be established in this area. The local public health unit has been underreporting the presence of B. burgdorferi s.l.-infected I. scapularis in the area encompassing Dundas. Our findings raise concerns about the need to erect tick warning signs in parkland areas. Veterinarians, medical professionals, public health officials, and the general public must be vigilant that Lyme disease-carrying blacklegged ticks pose a public health risk in the Dundas area and the surrounding Hamilton-Wentworth region.

The clinical differentiation of progressive supranuclear palsy from Parkinson's disease can be challenging, due to overlapping clinical features and a lack of diagnostic markers. Abnormalities in cognitive function form part of the clinical spectrums of these diseases and distinctive cognitive profiles may be helpful in differentiating these diseases in the diagnostic period. A comprehensive neuropsychological test battery was administered to 12 patients with clinically diagnosed progressive supranuclear palsy and 12 patients with Parkinson's disease matched for age and disease duration. Effect size (Cohen's d) was calculated for cognitive tests that were significantly different between groups. Patients with progressive supranuclear palsy performed significantly worse than those with Parkinson's disease on measures of processing speed, verbal fluency, planning, verbal abstract reasoning, verbal memory, and made more perseverative responses on a set shifting task. Measures of executive function, manual dexterity and processing speed were most diagnostically useful (Cohen's d > 2.0) in differentiating between progressive supranuclear palsy and Parkinson's disease. These findings suggest that more severe and prominent 'frontal' cognitive deficits in patients with progressive parkinsonism would be helpful in predicting progressive supranuclear palsy rather than Parkinson's disease and these findings may contribute to the development of diagnostic criteria., (© 2014 The British Psychological Society.)

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that can cause fever and chronic arthritis in humans. CHIKV that is generated in mosquito or mammalian cells differs in glycosylation patterns of viral proteins, which may affect its replication and virulence. Herein, we compare replication, pathogenicity, and receptor binding of CHIKV generated in Vero cells (mammal) or C6/36 cells (mosquito) through a single passage. We demonstrate that mosquito cell-derived CHIKV (CHIKV mos) has slower replication than mammalian cell-derived CHIKV (CHIKV vero), when tested in both human and murine cell lines. Consistent with this, CHIKV mos infection in both cell lines produce less cytopathic effects and reduced antiviral responses. In addition, infection in mice show that CHIKV mos produces a lower level of viremia and less severe footpad swelling when compared with CHIKV vero. Interestingly, CHIKV mos has impaired ability to bind to glycosaminoglycan (GAG) receptors on mammalian cells. However, sequencing analysis shows that this impairment is not due to a mutation in the CHIKV E2 gene, which encodes for the viral receptor binding protein. Moreover, CHIKV mos progenies can regain GAG receptor binding capability and can replicate similarly to CHIKV vero after a single passage in mammalian cells. Furthermore, CHIKV vero and CHIKV mos no longer differ in replication when N-glycosylation of viral proteins was inhibited by growing these viruses in the presence of tunicamycin. Collectively, these results suggest that N-glycosylation of viral proteins within mosquito cells can result in loss of GAG receptor binding capability of CHIKV and reduction of its infectivity in mammalian cells.

Cache Valley virus (CVV; Family Bunyavidae, Genus Orthobunyavirus) is a mosquito-borne zoonosis that frequently infects humans and livestock in North and Central America. In the northeastern United States, CVV transmission is unpredictable from year-to-year and may derive from the periodic extinction and reintroduction of new virus strains into this region. To evaluate this possibility, we sequenced and analyzed numerous CVV isolates sampled in Connecticut during an 18-year period to determine how the virus population may change over time. Phylogenetic analyses showed the establishment of a new viral lineage during 2010 that became dominant by 2014 and appears to have originated from southern Mexico. CVV strains from Connecticut also grouped into numerous sub-clades within each lineage that included viruses from other U.S. states and Canada. We did not observe the development and stable persistence of local viral clades in Connecticut, which may reflect the episodic pattern of CVV transmission. Together, our data support the emergence of a new lineage of CVV in the northeastern United States and suggest extensive dispersal of viral strains in North America., (© The American Society of Tropical Medicine and Hygiene.)