Exploring the biotechnological potential of Acinetobacter soli ANG344B: A novel bacterium for 2-phenylethanol production.

A bacterium, Acinetobacter soli ANG344B, isolated from river water, exhibited an exceptional capacity to produce 2-phenylethanol (2-PE) using L-phenylalanine (L-Phe) as a precursor-a capability typically observed in yeasts rather than bacteria. Bioreactor experiments were conducted to evaluate the production performance, using glucose as the carbon source for cellular growth and L-Phe as the precursor for 2-PE production. Remarkably, A. soli ANG344B achieved a 2-PE concentration of 2.35 ± 0.26 g/L in just 24.5 h of cultivation, exhibiting a global volumetric productivity of 0.10 ± 0.01 g/L.h and a production yield of 0.51 ± 0.01 g2-PE/gL-Phe, a result hitherto reported only for yeasts. These findings position A. soli ANG344B as a highly promising microorganism for 2-PE production. Whole-genome sequencing of A. soli strain ANG344 revealed a genome size of 3.52 Mb with a GC content of 42.7 %. Utilizing the Rapid Annotation using Subsystem Technology (RAST) server, 3418 coding genes were predicted, including genes coding for enzymes previously associated with the metabolic pathway of 2-PE production in other microorganisms, yet unreported in Acinetobacter species. Through gene mapping, 299 subsystems were identified, exhibiting 30 % subsystem coverage. The whole genome sequence data was submitted to NCBI GeneBank with the BioProject ID PRJNA982713. These draft genome data offer significant potential for exploiting the biotechnological capabilities of A. soli strain ANG344 and for conducting further comparative genomic studies.

Preclinical Efficacy of Cabazitaxel Loaded Poly(2-alkyl cyanoacrylate) Nanoparticle Variants.

Background: Biodegradable poly(alkyl cyanoacrylate) (PACA) nanoparticles (NPs) are receiving increasing attention in anti-cancer nanomedicine development not only for targeted cancer chemotherapy, but also for modulation of the tumor microenvironment. We previously reported promising results with cabazitaxel (CBZ) loaded poly(2-ethylbutyl cyanoacrylate) NPs (PEBCA-CBZ NPs) in a patient derived xenograft (PDX) model of triple-negative breast cancer, and this was associated with a decrease in M2 macrophages. The present study aims at comparing two endotoxin-free PACA NP variants (PEBCA and poly(2-ethylhexyl cyanoacrylate); PEHCA), loaded with CBZ and test whether conjugation with folate would improve their effect. Methods: Cytotoxicity assays and cellular uptake of NPs by flow cytometry were performed in different breast cancer cells. Biodistribution and efficacy studies were performed in PDX models of breast cancer. Tumor associated immune cells were analyzed by multiparametric flow cytometry. Results: In vitro studies showed similar NP-induced cytotoxicity patterns despite difference in early NP internalization. On intravenous injection, the liver cleared the majority of NPs. Efficacy studies in the HBCx39 PDX model demonstrated an enhanced effect of drug-loaded PEBCA variants compared with free drug and PEHCA NPs. Furthermore, the folate conjugated PEBCA variant did not show any enhanced effects compared with the unconjugated counterpart which might be due to unfavorable orientation of folate on the NPs. Finally, analyses of the immune cell populations in tumors revealed that treatment with drug loaded PEBCA variants affected the myeloid cells, especially macrophages, contributing to an inflammatory, immune activated tumor microenvironment. Conclusion: We report for the first time, comparative efficacy of PEBCA and PEHCA NP variants in triple negative breast cancer models and show that CBZ-loaded PEBCA NPs exhibit a combined effect on tumor cells and on the tumor associated myeloid compartment, which may boost the anti-tumor response.

Associations between PBMC whole genome transcriptome, muscle strength, muscle mass, and physical performance in healthy home-dwelling older women.

Increasing age is accompanied by many changes, including declining functional skeletal muscle health and immune dysfunction. Peripheral blood mononuclear cells (PBMCs) are circulating cells that assemble an immune response, but their whole genome transcriptome has not been studied in the context of age-related muscle health. Consequently, this article explored associations between three muscle variables indicative of functional muscle health - maximum handgrip strength (muscle strength), appendicular skeletal muscle mass index (ASMI, muscle mass), and gait speed (physical performance) - and two groups of bioinformatics-generated PBMC gene expression features (gene expression-estimated leukocyte subset proportions and gene clusters). We analyzed cross-sectional data from 95 home-dwelling healthy women ≥ 70 years, using "cell-type identification by estimating relative subsets of RNA transcripts" (CIBERSORT) to estimate leukocyte subset proportions and "weighted correlation network analysis" (WGCNA) to generate gene clusters. Associations were studied using linear regression models and relevant gene clusters were subjected to gene set enrichment analysis using gene ontology. Gait speed and ASMI associated with CIBERSORT-estimated monocyte proportions (ß = - 0.090, 95% CI = (- 0.146, - 0.034), p-value = 0.002 for gait speed, and ß = - 0.206, 95% CI = (- 0.385, - 0.028), p-value = 0.024 for ASMI), and gait speed associated with CIBERSORT-estimated M2 macrophage proportions (ß = - 0.026, 95% CI = (- 0.043, - 0.008), p-value = 0.004). Furthermore, maximum handgrip strength associated with nine WGCNA gene clusters, enriched in processes related to immune function and skeletal muscle cells (ß in the range - 0.007 to 0.008, p-values < 0.05). These results illustrate interactions between skeletal muscle and the immune system, supporting the notion that age-related functional muscle health and the immune system are closely linked.

A multi-centre study of learning styles of new nursing students.

AIMS: To describe beginning nursing student learning styles and to determine whether there are relationships between learning style and sociodemographic or educational background. We also aim to compare these data with the normative values of the 3.1 version of the Kolb learning style inventory and verify its reliability. BACKGROUND/LITERATURE: Learning style is related to the way in which an individual interacts with learning conditions, involving cognitive, affective, physical and environmental aspects. In nursing education, few studies have focused on knowing the learning styles of beginning nursing students, despite the need for students to develop critical thinking skills required of nurses. DESIGN: Descriptive, cross-sectional, correlational, multi-site study. METHOD: The study took place at five universities in Brazil and one university in the United States. Data were collected from a sample of beginning nursing students (n = 176) in 2019, using the Portuguese and English 3.1 versions of Kolb's learning style inventory. RESULTS: Most students' learning styles were classified as divergers (34.10%), while 28.41% were assimilators, 23.86% were accommodators and 13.63% were convergers. Kolb describes learning as a four-stage cyclical process grounded in experience. The mean score of one learning stage, the concrete experience, showed those who attended public high schools compared with private high schools was higher (mean = 26.22; p = .0019), and there were different results between Brazilian state universities, Brazilian federal universities and the American university (p = .0149). CONCLUSION: The diverger style was the most common among beginning nursing students in Brazil and the United States. There was a significant relationship between learning style, the educational background of students, the type of institution they attend and their previous experience with some active learning methodologies. RELEVANCE TO CLINICAL PRACTICE: Aligned with Kolb's experiential learning theory, nurse educators must be aware of student learning styles so they may use teaching strategies to meet their students' needs.

Loss of muscleblind splicing factor shortens Caenorhabditis elegans lifespan by reducing the activity of p38 MAPK/PMK-1 and transcription factors ATF-7 and Nrf/SKN-1.

Muscleblind-like splicing regulators (MBNLs) are RNA-binding factors that have an important role in developmental processes. Dysfunction of these factors is a key contributor of different neuromuscular degenerative disorders, including Myotonic Dystrophy type 1 (DM1). Since DM1 is a multisystemic disease characterized by symptoms resembling accelerated aging, we asked which cellular processes do MBNLs regulate that make them necessary for normal lifespan. By utilizing the model organism Caenorhabditis elegans, we found that loss of MBL-1 (the sole ortholog of mammalian MBNLs), which is known to be required for normal lifespan, shortens lifespan by decreasing the activity of p38 MAPK/PMK-1 as well as the function of transcription factors ATF-7 and SKN-1. Furthermore, we show that mitochondrial stress caused by the knockdown of mitochondrial electron transport chain components promotes the longevity of mbl-1 mutants in a partially PMK-1-dependent manner. Together, the data establish a mechanism of how DM1-associated loss of muscleblind affects lifespan. Furthermore, this study suggests that mitochondrial stress could alleviate symptoms caused by the dysfunction of muscleblind splicing factor, creating a potential approach to investigate for therapy.

Transtorno do espectro autista: capacitação de professores para atividades escolares em grupo / Autism spectrum disorder: teacher training for group school activities / Trastorno del espectro autista: capacitación de profesores para actividades escolares en grupo

Dentre as dificuldades encontradas para a inclusão de crianças com Transtorno do Espectro Autista (TEA) nas escolas está a falta de conhecimentos técnicos e instrumentos específicos que ofereçam suporte à prática dos profissionais da educação e às escolas. Considerando essa necessidade, foi aplicado um programa de capacitação, com base no referencial teórico da Análise do Comportamento, para 18 professores do Ensino Fundamental de uma escola de cidade do interior do Estado de São Paulo, com o objetivo de capacitá-los a aprenderem estratégias para promover a participação de alunos com TEA em atividades de grupo utilizando jogos cooperativos. Foram ministradas cinco aulas com enfoque prático, abordando os temas: jogos cooperativos, prompt e reforço. A avaliação do curso foi feita de três modos: aplicação de Pré-Teste, Pós-Teste e Follow-up. Para avaliar a aprendizagem dos professores participantes, foi solicitada aos professores uma atividade de simulação de situações e intervenções vivenciadas junto a alunos com TEA, a partir dos conteúdos aprendidos, e a aplicação da Escala de Avaliação do Ensino em Língua Oral em Contexto Escolar (EVALOE) para avaliar a atuação dos ministrantes. Os resultados indicaram que os participantes apresentaram mais acertos no pós-teste do que no pré-teste, a avaliação deles sobre o curso foi positiva e a atuação dos ministrantes satisfatória. Ressalta-se a importância de elaborar capacitações para os docentes e a necessidade de fornecer a eles mais materiais de apoio e atividades de autoaprendizagem.

Among the difficulties found for the inclusion of children with Autism Spectrum Disorder (ASD) in schools is the lack of technical knowledge and specific instruments that support the practice of education professionals and schools. Considering this need, a training program has been implemented based on the theoretical framework of behavior analysis for 18 elementary school teachers from a school in a city in the countryside of the State of São Paulo in order to enable them to learn strategies to promote the participation of students with ASD in group activities using cooperative games. Five classes have been given with a practical approach, addressing the themes: cooperative games, prompt and reinforcement. The evaluation of the course has been done in three steps: Pre-Test, Post-Test and Follow-up application. In order to evaluate the learning of the participating teachers, they performed a simulation activity of situations and interventions experienced with students with ASD based on the contents learned. In addition, the Assessment Scale of Oral Language Teaching at the School Context (EVALOE) was applied to evaluate the performance of the instructors. The results have indicated that the participants have had more correct answers in the Post-test than in the Pre-test, their evaluation of the course has been positive, and the performance of the students satisfactory. The importance of developing training for teachers and the need to provide them with more support materials and self-learning activities is emphasized.

En medio de las dificultades encontradas en la inclusión de los niños y niñas con Trastorno del Espectro Autista (TEA) en las escuelas, está la falta de conocimientos técnicos y instrumentos específicos que conceden soporte a la práctica de los profesionales de la educación y a las escuelas. Considerando esta necesidad, se aplicó un programa de capacitación a 18 maestros de la Escuela Primaria y Secundaria de una ciudad del estado de São Paulo, con el fin de capacitarles para promover la participación de alumnos con TEA en actividades en grupo utilizando los juegos cooperativos. Cinco clases fueron ministradas con un enfoque práctico, abordando los temas: juegos cooperativos, Prompt y refuerzo. Los resultados indicaron que los profesores tuvieron más éxito en el post-test que en el pre-test, aplicados para evaluar el curso, y su evaluación, en general, fue positiva. Se hace hincapié en la importancia de elaborar la capacitación de los docentes y la necesidad de proporcionarles más materiales de apoyo y actividades de autoaprendizaje.

Alpha-Terpineol as Antitumor Candidate in Pre-Clinical Studies.

BACKGROUND: Alpha-terpineol is monoterpene alcohol with anti-tumor activity against different tumor cell lines (lung, breast, leukemias and colorectal) through blockage of NF-kB expression, which play an important role in tumor cells growth. OBJECTIVE: Evaluate the antitumor activity of alpha-terpineol in murine Sarcoma 180 cell line. METHODS: For the tests, different cytotoxic and genotoxic assays were used, including Trypan blue, cytokinesis- blocked micronucleus assay, comet assay, agarose gel DNA fragmentation, flow cytometry and cell viability using fluorescence. Ascitic fluid cells from sarcoma 180 were obtained from Mus musculus peritoneal cavity and Alpha-terpineol was tested at 100, 250 and 500 µg/mL. Doxorubicin and Cisplatin were used as positive controls. RESULTS: Cytotoxic effects of alpha-terpineol were found in all concentrations tested, reducing cell viability in 50.9; 38.53; 30.82% at 100, 250 and 500 µg/mL, respectively. Alpha-terpineol induced genotoxic effects due to DNA fragmentation (increased frequency and index of damage), and was clastogenic by increased micronuclei formation, nucleoplasmic bridges and nuclear buds. DNA fragmentation and increased cell death indicated that alpha-terpineol can cause early, late, and necrotic apoptosis. CONCLUSION: Our data indicate that alpha-terpineol has antitumor activity revealed by cytogenetic mechanisms and / or loss of cell membrane integrity.

Feeding strategies to optimize vanillin production by Amycolatopsis sp. ATCC 39116.

The growing consumer demand for natural products led to an increasing interest in vanillin production by biotechnological routes. In this work, the biotechnological vanillin production by Amycolatopsis sp. ATCC 39116 is studied using ferulic acid as precursor, aiming to achieve maximized vanillin productivities. During biotech-vanillin production, the effects of glucose, vanillin and ferulic acid concentrations in the broth proved to be relevant for vanillin productivity. Concerning glucose, its presence in the broth during the production phase avoids vanillin conversion to vanillic acid and, consequently, increases vanillin production. To avoid the accumulation of vanillin up to a toxic concentration level, a multiple-pulse-feeding strategy is implemented, with intercalated vanillin removal from the broth and biomass recovery. This strategy turned out fruitful, leading to 0.46 g L-1 h-1 volumetric productivity of vanillin of and a production yield of 0.69 gvanillin gferulic acid-1, which are among the highest values reported in the literature for non-modified bacteria.

Augmented-limited regression models with an application to the study of the risk perceived using continuous scales.

Studies of risk perceived using continuous scales of [0,100] were recently introduced in psychometrics, which can be transformed to the unit interval, but the presence of zeros or ones are commonly observed. Motivated by this, we introduce a full inferential set of tools that allows for augmented and limited data modeling. We considered parameter estimation, residual analysis, influence diagnostic and model selection for zero-and/or-one augmented beta rectangular (ZOABR) regression models and their particular nested models, which is based on a new parameterization of the beta rectangular distribution. Different from other alternatives, we performed maximum-likelihood estimation using a combination of the EM algorithm (for the continuous part) and Fisher scoring algorithm (for the discrete part). Also, we perform an additional step, by considering other link functions, besides the usual logistic link, for modeling the response mean. By considering randomized quantile residuals, (local) influence diagnostics and model selection tools, we identified that the ZOABR regression model is the best one. We also conducted extensive simulations studies, which indicate that all developed tools work properly. Finally, we discuss the use of this type of models to treat psychometric data. It is worthwhile to mention that applications of the developed methods go beyond to Psychometric data. Indeed, they can be useful when the response variable in bounded, including or not the respective limits.

Red Propolis and Its Dyslipidemic Regulator Formononetin: Evaluation of Antioxidant Activity and Gastroprotective Effects in Rat Model of Gastric Ulcer.

Propolis has various pharmacological properties of clinical interest, and is also considered a functional food. In particular, hydroalcoholic extracts of red propolis (HERP), together with its isoflavonoid formononetin, have recognized antioxidant and anti-inflammatory properties, with known added value against dyslipidemia. In this study, we report the gastroprotective effects of HERP (50-500 mg/kg, p.o.) and formononetin (10 mg/kg, p.o.) in ethanol and non-steroidal anti-inflammatory drug-induced models of rat ulcer. The volume, pH, and total acidity were the evaluated gastric secretion parameters using the pylorus ligature model, together with the assessment of gastric mucus contents. The anti-Helicobacter pylori activities of HERP were evaluated using the agar-well diffusion method. In our experiments, HERP (250 and 500 mg/kg) and formononetin (10 mg/kg) reduced (p < 0.001) total lesion areas in the ethanol-induced rat ulcer model, and reduced (p < 0.05) ulcer indices in the indomethacin-induced rat ulcer model. Administration of HERP and formononetin to pylorus ligature models significantly decreased (p < 0.01) gastric secretion volumes and increased (p < 0.05) mucus production. We have also shown the antioxidant and anti-Helicobacter pylori activities of HERP. The obtained results indicate that HERP and formononetin are gastroprotective in acute ulcer models, suggesting a prominent role of formononetin in the effects of HERP.

Associations Between Season, Processing Plant, and Hide Cleanliness Scores with Prevalence and Concentration of Major Shiga Toxin-Producing Escherichia coli on Beef Cattle Hides.

The objectives of this study were (1) to estimate the prevalence and concentration of the seven major Shiga toxin-producing Escherichia coli (STEC) serogroups (O26, O45, O103, O111, O121, O145, and O157), collectively called STEC-7, on cattle hides collected in different seasons and beef processing plants; and (2) to determine associations of season, plant, and hide cleanliness scores with the prevalence and concentration of STEC-7. A total of 720 hide surface samples (240/season) were collected over three seasons (summer and fall 2015 and spring 2016) from beef cattle carcasses in four commercial processing plants in the United States. Samples were subjected to selective culture and spiral plating methods. Overall model-adjusted mean prevalence (95% confidence interval) was 0.3% (0.03-2.3%) for STEC O26; 0.05% (<0.01-8.5%) for STEC O45; 0.2% (0.02-1.9%) for STEC O103; 0.05% (<0.01-8.5%) for STEC O145; and 3.1% (0.6-15.2%) for STEC O157. Four percent of hide samples were enumerable for STEC O157; mean concentration (standard deviation) = 2.1 (0.7) log10 colony-forming units (CFUs)/100 cm2. No samples were enumerable for non-O157 STEC. Hide-on prevalence of STEC O157 and STEC non-O157 (specifically of STEC O103) was higher in summer and spring, respectively. Across seasons and plants, the most common STEC non-O157 serogroups in this study (O26 and O103) were associated with a higher prevalence of STEC O157. Season and plant played a role in prevalence and concentration of STEC in beef cattle hides, varying by serogroup. Tailoring mitigation strategies at the plant can be challenging and processors would benefit from supplementary preharvest interventions to reduce overall contamination pressure at the plant, especially in fall and spring months when hide-on prevalence of STEC non-O157 is higher.

Perspectives Regarding the Risk of Introduction of the Japanese Encephalitis Virus (JEV) in the United States.

Japanese encephalitis (JE) is a zoonotic, emerging disease transmitted by mosquito vectors infected with the Japanese encephalitis virus (JEV). Its potential for emergence into susceptible regions is high, including in the United States (US), and is a reason of economic concern among the agricultural community, and to public health due to high morbidity and mortality rates in humans. While exploring the complexities of interactions involved with viral transmission, we proposed a new outlook on the role of vectors, hosts and the environment under changing conditions. For instance, the role of feral pigs may have been underappreciated in our previous work, given research keeps pointing to the importance of susceptible populations of wild swine in naïve regions as key elements for the introduction of emergent vector-borne diseases. High risk of JEV introduction has been associated with the transportation of infected mosquitoes via aircraft. Nonetheless, no JEV outbreaks have been reported in the US to date and results from a qualitative risk assessment considered the risk of establishment to be negligible under the current conditions (environmental, vector, pathogen, and host). In this work, we discuss virus-vector-host interactions and ecological factors important for virus transmission and spread, review research on the risk of JEV introduction to the US considering the implications of risk dismissal as it relates to past experiences with similar arboviruses, and reflect on future directions, challenges, and implications of a JEV incursion.

Assessment of the use of Unna boot in the treatment of chronic venous leg ulcers in adults: systematic review protocol.

INTRODUCTION: Chronic venous insufficiency (CVI) is an anomaly of the normal functioning of the venous system caused by valvular incompetence with or without the obstruction of venous flow. This condition can affect either or both of the superficial and the deep venous systems. Venous dysfunction can even result in congenital or acquired disorders, and its complications include venous leg ulcers (VLUs). The objective of this systematic review is to determine the effectiveness of Unna boot in the treatment of wound healing of VLU by assessing the quality of the available evidence. METHODS AND ANALYSIS: A literature search in PubMed, CINAHL, Scopus, Web of Science, Cochrane Library, BVS/BIREME, Embase, ProQuest, BDTD, Thesis and Dissertation Catalog, Sao Paulo Research Foundation/Thesis and dissertation, OPEN THESIS, A service of the US National Institute of Health, Center for Reviews and Dissemination-University of New York and SciElo published in the last 10 years, the period from January 1999 to March 2019. The review will include primary studies (original), and Controlled Trials or Observational studies (cross-sectional, case-control or longitudinal studies) with VLU. The exclusion will include leg ulceration due to different causes, such as pressure, arterial, diabetic or mixed-aetiology leg ulcers. Data synthesis will be performed using a narrative summary and quantitative analysis. ETHICS AND DISSEMINATION: This systematic review does not require approval by the ethics committee, as individual patient data will not be collected. Dissemination of findings will be through publications in peer-reviewed journals and/or via conference presentations. PROSPERO REGISTRATION NUMBER: CRD42019127947.

Quantification of Bacteria Indicative of Fecal and Environmental Contamination from Hides to Carcasses.

Fecal bacteria, which reside in the gastrointestinal tract of cattle, can contaminate beef carcasses during processing. In beef cattle slaughter plants, the presence and concentrations of generic Escherichia coli, coliforms, Enterobacteriaceae (EB), and total aerobic bacteria are monitored as indicator organisms of fecal and environmental contamination. The objectives of this study were as follows: (1) to determine the concentrations of generic E. coli, coliforms, EB, and aerobic bacteria on beef carcasses at different processing points in Midwestern commercial beef slaughter plants during the summer, spring, and fall seasons; and (2) to estimate bacterial transfer on carcasses during the hide removal and evisceration processes. Hide and carcass surface sample swabs were collected from slaughtered cattle at four large commercial processing plants. At each plant visit (3 visits to each of the 4 plants) and during 3 seasons, 20 samples were collected at 5 points: hide-on (hide of animal near exsanguination pit), hide-off carcass, pre-evisceration carcass, postevisceration carcass, and postintervention carcass, for a total of 3600 samples. Bacterial concentrations were determined using 3M™ Petrifilm™ plates. Associations between season and processing plant with concentrations of E. coli, coliforms, EB, and total aerobic bacteria, overall, between hide-on and hide-off, and between pre- and post-evisceration, were evaluated using multilevel mixed-effects linear regression models. Bacterial concentrations on beef carcasses significantly decreased throughout processing. Moreover, hide removal was an important source of carcass contamination, given bacterial concentrations detected on hide-off carcass samples were the highest, and bearing in mind that carcass muscle surfaces should be sterile. Results from this study indicate that the interventions applied by the processing plants were effective, as they probably contributed to the significant reduction of bacterial concentrations of carcasses.

Introduction of the Japanese encephalitis virus (JEV) in the United States - A qualitative risk assessment.

The purpose of this risk assessment (RA) was to qualitatively estimate the risk of emergence of the Japanese encephalitis virus (JEV) in the United States (US). We followed the framework for RA of emerging vector-borne livestock diseases (de Vos et al. 2011), which consists of a structured questionnaire, whose answers to questions can be delivered in risk categories, descriptive statements, or yes or no type of answers, being supported by the literature. The most likely pathways of introduction of JEV identified were: (a) entry through infected vectors (by aircraft, cargo ships, tires, or wind); (b) import of infected viremic animals; (c) entry of viremic migratory birds; (d) import of infected biological materials; (e) import of infected animal products; (f) entry of infected humans; and (g) import/production of contaminated biological material (e.g., vaccines). From these pathways, the probability of introduction of JEV through infected adult mosquitoes via aircraft was considered very high and via ships/containers was deemed low to moderate. The probability of introduction via other pathways or modes of entry (vector eggs or larvae, hosts, and vaccines) was considered negligible. The probability of transmission of JEV was variable, ranging from low to high (in the presence of both competent vectors and hosts), depending on the area of introduction within the US. Lastly, the probability of establishment of JEV in the continental US was considered negligible. For that reason, we stopped the risk assessment at this point of the framework. This RA provides important information regarding the elements that contribute to the risk associated with the introduction of JEV in the US. This RA also indicates that infected mosquitoes transported in aircraft (and cargo ships) are the most likely pathway of JEV entry and therefore, mitigation strategies should be directed towards this pathway.

A quantitative risk assessment (QRA) of the risk of introduction of the Japanese encephalitis virus (JEV) in the United States via infected mosquitoes transported in aircraft and cargo ships.

Following a qualitative risk assessment, in which we identified and assessed all viable pathways for the introduction of the Japanese encephalitis virus (JEV) into the United States (US), we identified entry through infected vectors via aircraft and cargo ships as the most likely pathway, and thus considered it further in a quantitative risk assessment (QRA) model. The objective of this study was to evaluate the risk of introduction of JEV in the US via infected mosquitoes transported in aircraft and cargo ships arriving from Asia, using a QRA model. We created a stochastic model to quantify the probability of introduction of at least one infected mosquito in the continental US via aircraft and cargo ships, per at-risk period (March to October) or year, respectively. We modeled the following parameters: number of flights (per at-risk period, i.e., March to October) and cargo ships (per year) and per region, number of mosquitoes per flight and ship, number of mosquitoes that were not found and sensitivity of the mosquito collection method in aircraft, mosquito infection rates, and number of mosquitoes coming in aircraft per at-risk period (March to October) and cargo ships per year. Flight and cargo ship data pertained to years 2010-2016. For model building purposes, we only considered port-to-port vessels arriving from Asia to the US, we assumed that mosquitoes survive the trans-Pacific Ocean ship crossing and that the number of mosquitoes in cargo and passenger flights is similar. Our model predicted a very high risk (0.95 median probability; 95% CI = 0.80-0.99) of at least one infected mosquito being introduced in the US during the at-risk period, i.e., March to October, via aircraft transportation from JEV-affected countries in Asia. We also estimated that a median of three infected mosquitoes can enter the US during the at-risk period, i.e., March to October (95% CI = 1-7). The highest probability of introduction via aircraft was attributed to the Mediterranean California ecoregion (0.74; 95% CI = 0.50-0.90). We predicted, however, a negligible risk (0; 95% CI = 0.00-0.01) of at least one infected mosquito being introduced via cargo ships. Although the risk of introduction of JEV-infected mosquitoes by cargo ships was negligible, the risk via aircraft was estimated to be high. Our findings indicate the need to prioritize JEV prevention and control methods for aircraft-based pathways, such as aircraft disinfection. The quantitative estimates provided in this study are of interest to public health entities and other stakeholders, as they may support future interventions for preventing JEV introduction, as well as other vector-borne diseases, in the US and other countries.

Assessment of data on vector and host competence for Japanese encephalitis virus: A systematic review of the literature.

Japanese encephalitis virus (JEV) is a virus of the Flavivirus genus that may result in encephalitis in human hosts. This vector-borne zoonosis occurs in Eastern and Southeastern Asia and an intentional or inadvertent introduction into the United States (US) would have major public health and economic consequences. The objective of this study was to gather, appraise, and synthesize primary research literature to identify and quantify vector and host competence for JEV, using a systematic review (SR) of the literature. After defining the research question, we performed a search in selected electronic databases and journals. The title and abstract of the identified articles were screened for relevance using a set of exclusion and inclusion criteria, and relevant articles were subjected to a risk of bias assessment, followed by data extraction. Data were extracted from 171 peer-reviewed articles. Most studies were observational studies (59.1%) and reported vector competence (60.2%). The outcome measures reported pertained to transmission efficiency, host preference, and vector susceptibility to infection within vector competence; and susceptibility to infection within host competence. Regarding vector competence, the proportion of JEV infection reported across all 149 mosquito species in all observational studies ranged from 0 to 100%. In experimental studies, infection, dissemination, and transmission rates varied between 0 and 100%. Minimum infection rates (MIR) varied between 0 and 333.3 per 1000 mosquitoes. Maximum likelihood estimation (MLE) values ranged from 0 to 53.8 per 1000 mosquitoes. The host species in which mosquitoes mostly fed consisted of pigs and cattle (total of 84 blood meals taken by mosquitoes from each of these host species). As for host competence, the proportion of JEV infection varied between 0 (in rabbits, reptiles, and amphibians) and 88.9% (cattle). This SR presents comprehensive data on JEV vector and host competence, which can be used to quantify risks associated with the introduction of JEV into the US.

Meta-Analyses of Japanese Encephalitis Virus Infection, Dissemination, and Transmission Rates in Vectors.

The objective of this work was to summarize and quantify Japanese encephalitis virus (JEV) infection, dissemination, and transmission rates in mosquitoes, using a meta-analysis approach. Data were obtained from experimental studies, gathered by means of a systematic review of the literature. Random-effects subgroup meta-analysis models by mosquito species were fitted to estimate pooled estimates and to calculate the variance between studies for three outcomes of interest: JEV infection, dissemination, and transmission rates in mosquitoes. To identify sources of heterogeneity among studies and to assess the association between different predictors (mosquito species, virus administration route, incubation period, and diagnostic method) with the outcome JEV infection rate in vectors, we fitted univariable meta-regression models. Mosquito species and administration route represented the main sources of heterogeneity associated with JEV infection rate in vectors. This study provided summary effect size estimates to be used as reference for other investigators when assessing transmission efficiency of vectors and explored sources of variability for JEV infection rates in vectors. Because transmission efficiency, as part of vector competence assessment, is an important parameter when studying the relative contribution of vectors to JEV transmission, our findings contribute to further our knowledge, potentially moving us toward more informed and targeted actions to prevent and control JEV in both affected and susceptible regions worldwide.

Meta-analyses of the proportion of Japanese encephalitis virus infection in vectors and vertebrate hosts.

BACKGROUND: Japanese encephalitis (JE) is a zoonosis in Southeast Asia vectored by mosquitoes infected with the Japanese encephalitis virus (JEV). Japanese encephalitis is considered an emerging exotic infectious disease with potential for introduction in currently JEV-free countries. Pigs and ardeid birds are reservoir hosts and play a major role on the transmission dynamics of the disease. The objective of the study was to quantitatively summarize the proportion of JEV infection in vectors and vertebrate hosts from data pertaining to observational studies obtained in a systematic review of the literature on vector and host competence for JEV, using meta-analyses. METHODS: Data gathered in this study pertained to three outcomes: proportion of JEV infection in vectors, proportion of JEV infection in vertebrate hosts, and minimum infection rate (MIR) in vectors. Random-effects subgroup meta-analysis models were fitted by species (mosquito or vertebrate host species) to estimate pooled summary measures, as well as to compute the variance between studies. Meta-regression models were fitted to assess the association between different predictors and the outcomes of interest and to identify sources of heterogeneity among studies. Predictors included in all models were mosquito/vertebrate host species, diagnostic methods, mosquito capture methods, season, country/region, age category, and number of mosquitos per pool. RESULTS: Mosquito species, diagnostic method, country, and capture method represented important sources of heterogeneity associated with the proportion of JEV infection; host species and region were considered sources of heterogeneity associated with the proportion of JEV infection in hosts; and diagnostic and mosquito capture methods were deemed important contributors of heterogeneity for the MIR outcome. CONCLUSIONS: Our findings provide reference pooled summary estimates of vector competence for JEV for some mosquito species, as well as of sources of variability for these outcomes. Moreover, this work provides useful guidelines when interpreting vector and host infection proportions or prevalence from observational studies, and contributes to further our understanding of vector and vertebrate host competence for JEV, elucidating information on the relative importance of vectors and hosts on JEV introduction and transmission.