Wastewater based epidemiology as a surveillance tool during the current COVID-19 pandemic on a college campus (East Carolina University) and its accuracy in predicting SARS-CoV-2 outbreaks in dormitories.

The COVID-19 outbreak led governmental officials to close many businesses and schools, including colleges and universities. Thus, the ability to resume normal campus operation required adoption of safety measures to monitor and respond to COVID-19. The objective of this study was to determine the efficacy of wastewater-based epidemiology as a surveillance method in monitoring COVID-19 on a college campus. The use of wastewater monitoring as part of a surveillance program to control COVID-19 outbreaks at East Carolina University was evaluated. During the Spring and Fall 2021 semesters, wastewater samples (N = 830) were collected every Monday, Wednesday, and Friday from the sewer pipes exiting the dormitories on campus. Samples were analyzed for SARS-CoV-2 and viral quantification was determined using qRT-PCR. During the Spring 2021 semester, there was a significant difference in SARS-CoV-2 virus copies in wastewater when comparing dorms with the highest number student cases of COVID-19 and those with the lowest number of student cases, (p = 0.002). Additionally, during the Fall 2021 semester it was observed that when weekly virus concentrations exceeded 20 copies per ml, there were new confirmed COVID-19 cases 85% of the time during the following week. Increases in wastewater viral concentration spurred COVID-19 swab testing for students residing in dormitories, aiding university officials in effectively applying COVID testing policies. This study showed wastewater-based epidemiology can be a cost-effective surveillance tool to guide other surveilling methods (e.g., contact tracing, nasal/salvia testing, etc.) to identify and isolate afflicted individuals to reduce the spread of pathogens and potential outbreaks within a community.

Development of novel compact wind tunnel for testing efficacy of insecticide formulated products in mosquitoes.

BACKGROUND: Ultra-low volume (ULV) space sprays aerosolize insecticide formulated products (FP) to contact flying mosquitoes, while barrier sprays expose mosquitoes to FP residue on vegetation and other surfaces. Centers for Disease Control and Prevention bottle bioassays used to assess insecticide resistance are based on residual active ingredient (AI) exposure and do not directly relate to FP efficacy. The current pilot study developed a novel compact wind tunnel for mosquito exposure to FP. Caged Aedes albopictus and Culex pipiens/quinquefasciatus were exposed to undiluted Biomist®3 + 15 FP (permethrin AI) or air (control) within the wind tunnel, transferred to new cages, and held in a 28 °C incubator. Separate mosquitoes were exposed to residual permethrin AI (8 µg mL-1 ) in bottle bioassays. Mortality was monitored 15, 30, 60, and 120 min post-exposure. RESULTS: Chi-square tests (P < 0.05) showed significantly higher mortality in Aedes compared to Culex populations for most time points in both bioassay and wind tunnel exposure groups. As expected, mosquitoes exposed to Biomist®3 + 15 showed higher mortality rates than bottle bioassay exposure to permethrin. Two Culex colonies resistant to permethrin in bottle bioassays were susceptible to Biomist®3 + 15 in the wind tunnel. CONCLUSION: The novel compact wind tunnel developed here may be an alternative to field trials for testing FP efficacy, avoiding factors such as weather, logistical planning, and extended personnel hours. The wind tunnel could allow programs to conveniently test efficacy of multiple FP. Comparisons of different insecticide exposure methods provide practical information to inform operational decisions. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Assessment of barrier treatments impacting Aedes albopictus (diptera: culicidae) using lambda-cyhalothrin and pyriproxyfen in a suburban neighborhood in Eastern North Carolina, 2018.

Pyrethroids are commonly used in barrier treatments but less is known about how mosquitoes are affected by the simultaneous application of an insect growth regulator (i.e., pyriproxyfen) used in barrier treatments. This field study, conducted from May 14 - October 16, 2018, evaluated the effect of lambda-cyhalothrin (pyrethroid adulticide) and pyriproxyfen on the reproduction (measured by fecundity and adult emergence) and abundance of Aedes albopictus. Nine properties were treated with Demand®CS 0.06% + Archer® 0.010% (every 60 days) and three control properties received no treatment (N=12 total properties). No significant (P>0.05) differences were observed between abundance of Ae. albopictus in treatment compared to control groups. However, significant differences were observed in abundance of Ae. albopictus adults between weeks at both control (P = 0.003) and treatment (P < 0.0001) properties. Results from our research show that continued studies are needed to determine the efficacy of barrier treatments for this species. .

Retrospective chart review of substance abuse in patients with psychiatric emergencies in an emerging urban county.

Objectives: Substance abuse is common in patients with psychiatric emergencies. To further understand the connection between substance abuse and psychiatric disorders, a retrospective chart review was done that included positive drug screens among patients with psychiatric emergencies and to determine whether there was an association between substances used and the psychiatric diagnosis. Methods: A retrospective chart review of patients seen in an emergency department with psychiatric emergencies was conducted. The review comprised 1000 charts with diagnoses of anxiety, depression, schizophrenia, attention-deficit/hyperactivity disorder (ADHD), bipolar disorder, alcohol abuse, or schizoaffective disorder. Data collected included patient demographics, tobacco abuse, chief complaint, arrival mode, voluntary versus involuntary status, suicide attempt on presentation, psychiatric diagnoses, urine drug screen, and ethanol results. Chi-square statistical analysis was conducted to examine the relationship between substances of abuse and psychiatric diagnoses. Results: Approximately 58% of patients with a history of psychiatric illness had a positive urine drug screen. Of 245 patients with schizoaffective disorder, 69 (28%) were positive for tetrahydrocannabinol (THC) and 48 (20%) were positive for cocaine. Of 225 patients with depression, 59 (29%) were positive for THC and 33 (15%) were positive for cocaine. Cannabis was the most commonly reported substance used among patients with depression, schizophrenia, anxiety, schizoaffective disorder, and bipolar disorder, and ethanol was most common in patients with ADHD. No significant correlations were found between psychiatric diagnosis and positive drug screens. A statistically significant secondary end point was found that White people using cannabinoids were more likely to attempt suicide than were African American people (P = 0.02). Conclusions: Positive drug screens were common among patients presenting to an ED with psychiatric emergencies. Cannabis was the most commonly reported substance used among patients independent of diagnosis. Ethanol was the most common in patients with ADHD. Urine drug screens are unlikely to provide insights into relationships between specific substance use and psychiatric emergencies.

Evaluation of surface versus total permethrin content in permethrin-treated clothing: Implications for protection against mosquitoes.

Permethrin is a synthetic pyrethroid insecticide used to treat uniforms (e.g. military, foresters) and other clothing to protect people against pests (e.g., mosquitoes, ticks). Pests contacting the surface of permethrin-treated clothing (PTC) are repelled and/or killed, depending on the dose and duration of exposure. Hence, it is important to assess the amount of permethrin on the surface of clothing. Fabric swatches prepared using two commercially available permethrin treatments (Insect Shield® & Sawyer Repellant) and one laboratory created treatment (4g permethrin/L) were tested. A Martindale Abrasion and Pilling Tester (MAPT) and gas chromatography were used to assess surface permethrin content (SPC) and total permethrin content (TPC). Sawyer PTC had the highest SPC (mean ± standard error) (32.68±14.55µg/g), followed by Insect Shield® (23.35±2.71µg/g) and lab-created 4g/L permethrin (8.7±0.78µg/g). SPC (after 1000 rubs on MAPT) for Insect Shield®, Sawyer, and 4g/L permethrin groups was significantly lower than TPC (P=0.011, P<0.001, and P=0.001, respectively). The SPC/TPC relationship varied widely between permethrin treatment methods and practical implications for this are discussed. Mosquito repellency tests indicate that Insect Shield®, Sawyer, and lab-created (4g/L) permethrin-treated fabrics showed a significantly higher repellency rate than control (untreated) fabrics (P=0.001, P<0.0001, P<0.0001, respectively). While Insect Shield® had the highest repellency rate against susceptible (53%) and exposed (48%) mosquitoes, differences between groups were not significant. Repellency rates indicate SPC plus other factors (e.g. treatment method) may contribute to mosquito repellency and should be considered in risk assessments for protection against pests.

Assessing Durability and Safety of Permethrin Impregnated Uniforms Used by Outdoor Workers to Prevent Tick Bites after One Year of Use.

Long lasting permethrin-impregnated (LLPI) clothing can retain permethrin and repel ticks for up to three months and without exceeding EPA-approved safe levels; however, little is known about longer term effects of wearing LLPI clothing. Here, permethrin content was measured in new forester pants soon after initial impregnation (Insect Shield) and again one year later after being repeatedly worn by foresters in the field. Urine samples were collected from foresters for biomonitoring of permethrin metabolites at multiple time intervals (pre-use, one-month, three-to-four-months, and one-year post-use). Lethality against nymphal Ixodes scapularis Say was measured in clothing after one year of wear by foresters. Furthermore, to test potential variability in permethrin impregnation of different batches of clothing, separate sets of clothing were anonymously sent to Insect Shield for permethrin treatment over a period of three months and permethrin was quantified. Results demonstrated 33% of participants' pants had no measurable permethrin after one year of wear and permethrin content and tick mortality varied significantly between clothing. Only two of the participants' clothing resulted in ≥ 30% tick mortality after one year of wear. Significant differences were observed in 3-PBA and trans-DCCA, but not cis-DCCA metabolites in participants over the four measured time points and were higher than general United States population levels. This study provides practical information on the safety (measured by urinary metabolites) over time of LLPI clothing. It also provides snapshots (pre-washing and after one year of wear) of effectiveness of LLPI clothing as personal protective equipment against ticks for outdoor workers.

Risk assessment of inhaled diacetyl from electronic cigarette use among teens and adults.

Diacetyl (C4H6O2) is a toxicant commonly found in electronic cigarettes (e-Cigs) as a flavoring component and an enhancer of e-juices. Lung injury in current and former workers in popcorn manufacturing suggests a possible association with diacetyl inhalation exposure. Although the number of e-Cig users continues to rise steadily among the teens and adults, the potential risk of pulmonary disease has not been characterized. A systematic review of the open literature identified bronchiolitis obliterans-a pathological inflammation resulting in fibrosis of the bronchioles leading to an irreversible limitation to airflow in lungs-as the primary outcome of diacetyl exposures. Following the deterministic United States National Research Council/Environmental Protection Agency's risk assessment framework, that consists of four key steps: hazard identification, dose-response assessment, exposure assessment and risk characterization, we estimated noncarcinogenic (systemic) risks using a Hazard Quotient (HQ) approach upon exposure to diacetyl among teens and adults who use e-Cigs. Based on the NIOSH Benchmark Dose (BMD; 0.0175 mg/kg-day) and modelled Average Daily Doses (ADDs; range 0.11-5.2 mg/kg-day), we estimated 12 different HQ values-a measure of non-carcinogenic risk for diacetyl inhalation exposures-all of which were greater than 1 (range 6.2875-297.1429), suggesting a significantly higher non-carcinogenic risk from diacetyl exposures among the teens and adults who use e-Cigs. These results underscore the need to regulate e-Cigs to protect teens and adults from diacetyl exposures and risk of developing lung injuries, including bronchiolitis obliterans.

Filtration efficiency of surgical sterilization fabric for respiratory protection during COVID-19 pandemic.

BACKGROUND: Due to COVID-19 and high demand for respirators, some healthcare professionals have been using the Halyard H600 fabric as an alternative to N95 respirators without testing the filtration efficiency of the fabric with established scientific methods. The purpose of this study was to assess the efficiency of the Halyard H600 as a respirator filtering material as compared to the NIOSH-certified N95 and P100 filters, and determine if H600 is a good alternative for respiratory protection for healthcare professionals during the COVID-19 pandemic. METHODS: Three filter types (Halyard H600, N95, and P100) were challenged with salt particles inside an exposure chamber at a flow rate of 43 LPM and relative humidity of 40 ± 2%. N95 and P100 respirator filters were tested initially to establish the validity of the chamber, followed by the Halyard H600 fabric. Particle penetration was measured using an aerosol spectrometer. The filtration efficiency was calculated for different particle sizes by measuring the particle number concentration upstream and downstream of the filter. The pressure drop across the filter materials was measured using a manometer. RESULTS: The efficiency of the P100 for particles ≥250 nm was 100%. The N95 efficiency was 97 ± 1% at 275 nm, 99 ± 0% at 324 nm, and 100% for larger particles. The Halyard H600 fabric had a variable efficiency with an average of 62 ± 28% at 275 nm, 89 ± 8% at 324 nm, and 100% efficiency for particles >450 nm. The pressure drop values for P100 and N95 were 32 and 8 mmH2O, respectively. The Halyard H600 fabric resistance increased dramatically from 30 mmH2O at the start of the exposure to 65 mmH2O after 16-minutes of exposure. CONCLUSION: The high variability in filter efficiency for particles ≤324 nm and the increased fabric breathing resistance demonstrate that the Halyard H600 has an inferior performance and is not a good substitute for N95 and P100. Thus, the use of the Halyard H600 fabric for respiratory protection is not recommended.

Mosquito-infecting virus Espirito Santo virus inhibits replication and spread of dengue virus.

The primary vector of dengue virus (DENV) is Aedes aegypti. The mosquito-infecting virus, Espirito Santo virus (ESV), does not infect Vero (mammalian) cells and grows in mosquito (C6/36) cells without cytopathic effects. Effects of ESV infection on replication of DENV were explored in vitro and in vivo, analyzing protein, RNA genome expression, and plaque formation. ESV and DENV simultaneous coinfection did not block protein synthesis from either virus but did result in inhibition of DENV replication in mosquito cells. Furthermore, ESV superinfected with DENV resulted in inhibition of DENV replication and spread in A. aegypti, thus reducing vector competence. Tissue culture experiments on viral kinetics of ESV and DENV coinfection showed that neither virus significantly affects the replication of the other in Vero, HeLa, or HEK cells. Hence, ESV blocks DENV replication in insect cells, but not the mammalian cells evaluated here. Our study provides new insights into ESV-induced suppression of DENV, a globally important pathogen impacting public health.

Assessing Insecticide Resistance in Adult Mosquitoes: Perspectives on Current Methods.

Mosquito insecticide resistance (IR) is a growing global issue that must be addressed to protect public health. Vector control programs (VCPs) should regularly monitor local mosquito populations for IR and plan control measures accordingly. In some cases, state/federal resources financially support this testing with expertise and/or training programs. Standardization of methods (eg, Centers for Disease Control and Prevention bottle bioassay, World Health Organization tube testing, dose-mortality bioassay) for monitoring IR must be prioritized. One solution is regional hubs of IR monitoring at the state or other level. Training programs on methodology and interpretation of results should be developed and routinely offered to local VCPs conducting IR testing in mosquitoes. Here, current methods for assessing mosquito IR are discussed and insights into a variety of questions from VCPs are considered. It is critical that methods for IR monitoring and data interpretation are standardized through routine training, with the goal of evidence-driven decision making to improve control of mosquitoes and mosquito-borne disease.

Bioabsorption and effectiveness of long-lasting permethrin-treated uniforms over three months among North Carolina outdoor workers.

BACKGROUND: Vector-borne diseases are an important cause of morbidity and mortality in the USA. Effective, convenient prevention methods are needed. Long-lasting permethrin-impregnated (LLPI) clothing can prevent tick bites, however, additional information is needed on the real-world effectiveness and safety of this preventative measure. METHODS: In this pilot study, we recruited state and county park employees from North Carolina to wear LLPI uniforms for three months during the summer of 2016. We collected spot urine samples for biomonitoring of permethrin metabolites at one week, one month and three months after first use of the LLPI uniform. Following three months of wear, we collected pants and socks and analyzed them for permethrin content and mortality to ticks and mosquitoes. RESULTS: Thirteen park employees were included in the analysis. Bioactive amounts of permethrin remained in all clothing swatches tested, although there was great variability. Tick mortality was high, with 78% of pant and 88% of sock swatches having mean knockdown percentages ≥ 85%. In contrast, mosquito mortality was low. Over the study period, the absorbed dosage of permethrin averaged < 4 µg/kg/d of body weight based on measurements of three metabolites. CONCLUSIONS: LLPI clothing retained permethrin and bioactivity against ticks after three months of use in real-world conditions. The estimated absorbed dosage of permethrin was well below the U.S. EPA level of concern, suggesting that LLPI clothing can be used safely by outdoor workers for tick bite prevention.

Evaluation of Insecticide Resistance in Aedes albopictus (Diptera: Culicidae) in North Carolina, 2017.

Mosquitoes may develop resistance to insecticide active ingredients (AIs) found in formulated products (FPs) due to environmental exposure from insecticides in mosquito control and/or unrelated to mosquito control, e.g., agricultural, household pest control. Mosquito control programs should implement resistance management strategies by assessing resistance in targeted populations, rotating different classes of insecticides based on resistance testing, and/or increasing insecticide concentration (i.e., saturation, using maximum labeled rate) to overcome emerging resistance. Resistance testing is often done solely on AIs, but should, in some cases, include both AIs and FPs at the concentrations mosquitoes may encounter in the field. The resistance/susceptibility status was determined for adulticides used in mosquito control. Centers for Disease Control and Prevention (CDC) bottle bioassays were used to assess resistance/susceptibility status for eight AIs (i.e., bifenthrin, permethrin, sumethrin/prallethrin, deltamethrin, tau-fluvalinate, chlorpyrifos, malathion, and naled) and eight FPs (TalStar, Biomist 3 + 15, Duet, Suspend Polyzone, Mavrik, MosquitoMist, Fyfanon, and Dibrom) that respectively contain the AIs. Current CDC guidelines were utilized: susceptible (97-100% mortality at diagnostic time [DT]), developing resistance (90-96% mortality at DT), or resistant (<90% mortality at DT). Significant differences were observed in mosquito susceptibility/resistance among and between AIs and FPs.

Impact of Mosquito Age and Insecticide Exposure on Susceptibility of Aedes albopictus (Diptera: Culicidae) to Infection with Zika Virus.

Zika virus (ZIKV) is primarily transmitted to humans by Aedes aegypti and Ae. albopictus. Vector⁻virus interactions influencing vector competence vary and depend on biological and environmental factors. A mosquito's chronological age may impact its immune response against virus infection. Insecticides, source reduction, and/or public education are currently the best defense against mosquitoes that transmit ZIKV. This study assessed the effects of a mosquito's chronological age at time of infection on its response to ZIKV infection. We exposed young (6⁻7 d post-emergence) and old (11⁻12 d post-emergence) Ae. albopictus to a sublethal dose of bifenthrin prior to oral exposure to blood meals containing ZIKV (7-day incubation period). Old mosquitoes experienced a significantly (p < 0.01) higher rate of mortality than young mosquitoes. Significantly higher ZIKV body titers (p < 0.01) were observed in the old control group compared to the young control group. Significantly higher (p < 0.01) ZIKV dissemination rates and leg titers (p < 0.01) were observed in old bifenthrin-exposed mosquitoes compared to old control mosquitoes or young bifenthrin-exposed or control mosquitoes. Hence, bifenthrin exposure may increase the potential for virus transmission; however, the degree of these impacts varies with mosquito age. Impacts of insecticides should be considered in risk assessments of potential vector populations.

Permethrin treated clothing to protect outdoor workers: evaluation of different methods for mosquito exposure against populations with differing resistance status.

Minimizing arthropod exposure (e.g. mosquito and tick bites) is vital to protect health of outdoor workers. Personal protective measures can help protect against exposure. Here, the quantity of permethrin was evaluated for different fabric types after washing. Cone and petri dish exposure assays were used to investigate the knockdown/mortality of permethrin-susceptible and permethrin-resistant populations of mosquitoes. Permethrin-treated clothing was effective against the tested mosquito population that was susceptible to permethrin but not a permethrin-resistant population. Permethrin quantity was significantly highest in the 100% cotton fabric and for the 0 wash group. Permethrin quantity in fabrics decreased with washing. No significant differences (p > 0.05) were observed in knockdown/mortality rates for either exposure method. The protective effect of permethrin-treated clothing against mosquitoes is impacted by many factors, e.g. wash frequency, fabric type, and the susceptibility/resistance status of local mosquito populations.

Insecticide Susceptibility Screening Against Culex and Aedes (Diptera: Culicidae) Mosquitoes From the United States.

Mosquitoes exposed to sublethal doses of insecticides may be selected for resistance to insecticide active ingredients (AIs). Mosquitoes are exposed to AIs through agricultural, public/private mosquito control programs, homeowners, and other sources. Hence, mosquito control programs should routinely measure the resistance/susceptibility status of mosquito populations of public health concern. The objectives here were to determine resistance status for six AIs used in adult mosquito control in the United States to assess how resistance/susceptibility differs between AI, mosquito species (states where > 1 species collected), and between years (some populations sampled for 2 yr). Field-collected eggs from 21 mosquito populations of six different species or hybrid species (Aedes albopictus Skuse [Diptera: Culicidae], Aedes aegypti L. [Diptera: Culicidae], Culex nigripalpus Theobald, Culex pipiens L. [Diptera: Culicidae], Culex quinquefasciatus Say [Diptera: Culicidae], Cx. pipiens/quinquefasciatus) were obtained. Centers for Disease Control and Prevention bottle bioassays were used to assess the resistance/susceptibility status for six AIs (bifenthrin, deltamethrin, etofenprox, malathion, permethrin, and phenothrin). World Health Organization guidelines were used to classify mosquitoes as susceptible (98-100% mortality at diagnostic time [DT]), possibly resistant (80-97% mortality at DT), or resistant (<80% mortality at DT). Significant differences were observed in mosquito susceptibility/resistance between species and AIs. In states where both Aedes and Culex were collected, the odds of exhibiting resistance in Culex were 68-69 times higher than Aedes (Texas odds ratio: 69.30; 95% confidence interval: 5.86, 819.44; P = 0.001; North Carolina odds ratio: 67.99; 95% confidence interval: 15.21, 303.94; P < 0.0001). Some level of resistance was detected against all tested AIs in several mosquito populations and some varied between 2015 and 2016.

Potential for sublethal insecticide exposure to impact vector competence of Aedes albopictus (Diptera: Culicidae) for dengue and Zika viruses.

Chikungunya, dengue, and Zika viruses (CHIKV, family Togaviridae, genus Alphavirus; DENV and ZIKV, family Flaviviridae, genus Flavivirus) are arboviruses that cause human epidemics. Due to the lack of vaccines for many mosquito-borne diseases, there is a need for mosquito control. In the US and other regions, residual barrier insecticide sprays are applied to foliage where female mosquitoes rest and/or sugar feed between blood meals. Residual sprays are an important control method for anthropogenic day-active sylvan mosquitoes such as Aedes albopictus (vector of CHIKV, DENV, and ZIKV) that are difficult to control using ultralow-volume sprays applied only at dusk or dawn when these mosquitoes are not active. In this exploratory study, we analyzed the extent to which ingestion of a sublethal dose of the active ingredient bifenthrin affected vector competence (i.e., infection, dissemination, and transmission) of Ae. albopictus for DENV and ZIKV. Two incubation periods (IPs; 7 and 14 d) were tested at 28°C for insecticide-fed and sugar-fed mosquitoes. We show that mosquitoes that were fed bifenthrin (0.128 µg/mL) mixed with sucrose solution exhibited significantly lower DENV infection rates and body titers after a 14-d IP. During the 7-d IP, one mosquito (sugar group) transmitted ZIKV. During the 14-d IP, 100% of mosquitoes showed body and leg ZIKV infections, and one mosquito (sugar+bifenthrin group) transmitted ZIKV. This is a preliminary communication, and more studies will be required to further investigate these findings. We expect the findings of this small-scale study to spur larger-scale investigations of the impacts of insecticides on mechanisms regulating vector competence, and exposure to other active ingredients, and aid in development of new insecticides.