SARS-CoV-2 wastewater variant surveillance: pandemic response leveraging FDA's GenomeTrakr network.

Wastewater surveillance has emerged as a crucial public health tool for population-level pathogen surveillance. Supported by funding from the American Rescue Plan Act of 2021, the FDA's genomic epidemiology program, GenomeTrakr, was leveraged to sequence SARS-CoV-2 from wastewater sites across the United States. This initiative required the evaluation, optimization, development, and publication of new methods and analytical tools spanning sample collection through variant analyses. Version-controlled protocols for each step of the process were developed and published on protocols.io. A custom data analysis tool and a publicly accessible dashboard were built to facilitate real-time visualization of the collected data, focusing on the relative abundance of SARS-CoV-2 variants and sub-lineages across different samples and sites throughout the project. From September 2021 through June 2023, a total of 3,389 wastewater samples were collected, with 2,517 undergoing sequencing and submission to NCBI under the umbrella BioProject, PRJNA757291. Sequence data were released with explicit quality control (QC) tags on all sequence records, communicating our confidence in the quality of data. Variant analysis revealed wide circulation of Delta in the fall of 2021 and captured the sweep of Omicron and subsequent diversification of this lineage through the end of the sampling period. This project successfully achieved two important goals for the FDA's GenomeTrakr program: first, contributing timely genomic data for the SARS-CoV-2 pandemic response, and second, establishing both capacity and best practices for culture-independent, population-level environmental surveillance for other pathogens of interest to the FDA. IMPORTANCE: This paper serves two primary objectives. First, it summarizes the genomic and contextual data collected during a Covid-19 pandemic response project, which utilized the FDA's laboratory network, traditionally employed for sequencing foodborne pathogens, for sequencing SARS-CoV-2 from wastewater samples. Second, it outlines best practices for gathering and organizing population-level next generation sequencing (NGS) data collected for culture-free, surveillance of pathogens sourced from environmental samples.

Tracking Vibrio: population dynamics and ecology of Vibrio parahaemolyticus and V. vulnificus in an Alabama estuary.

Vibrio is a genus of halophilic, gram-negative bacteria found in estuaries around the globe. Integral parts of coastal cultures often involve contact with vectors of pathogenic Vibrio spp. (e.g., consuming raw shellfish). High rates of mortality from certain Vibrio spp. infections demonstrate the need for an improved understanding of Vibrio spp. dynamics in estuarine regions. Our study assessed meteorological, hydrographic, and biological correlates of Vibrio parahaemolyticus and V. vulnificus at 10 sites in the Eastern Mississippi Sound System (EMSS) from April to October 2019. During the sampling period, median abundances of V. parahaemolyticus and V. vulnificus were 2.31 log MPN/L and 2.90 log MPN/L, respectively. Vibrio spp. dynamics were largely driven by site-based variation, with sites closest to freshwater inputs having the highest abundances. The E-W wind scalar, which affects Ekman transport, was a novel Vibrio spp. correlate observed. A potential salinity effect on bacterial-particle associations was identified, where V. vulnificus was associated with larger particles in conditions outside of their optimal salinity. Additionally, V. vulnificus abundances were correlated to those of harmful algal species that did not dominate community chlorophyll. Correlates from this study may be used to inform the next iteration of regionally predictive Vibrio models and may lend additional insight to Vibrio spp. ecology in similar systems. IMPORTANCE: Vibrio spp. are bacteria found in estuaries worldwide; some species can cause illness and infections in humans. Relationships between Vibrio spp. abundance, salinity, and temperature are well documented, but correlations to other environmental parameters are less understood. This study identifies unique correlates (e.g., E-W wind scalar and harmful algal species) that could potentially inform the next iteration of predictive Vibrio models for the EMSS region. Additionally, these correlates may allow existing environmental monitoring efforts to be leveraged in providing data inputs for future Vibrio risk models. An observed correlation between salinity and V. vulnificus/particle-size associations suggests that predicted environmental changes may affect the abundance of Vibrio spp. in certain reservoirs, which may alter which vectors present the greatest vibrio risk.

Future Doctors From Underrepresented in Medicine Communities at the University of Utah.

Background and Objectives: Future Doctors (FD), a high school pathway program, was developed to address the lack of compositional diversity in the health professions at our health sciences campus. Methods: We obtained, analyzed, and compared data on FD student demographic and educational achievement at undergraduate and graduate programs at the University of Utah and graduate programs at other institutions to non-FD students. We followed students from high school to graduate school. Results: We analyzed data from 1,897 FD participants (2014-2019). FD participants were 71% women, with 50.3% identifying as students of color. Ninety-eight students matriculated in graduate school, with 75 (76%) remaining at University of Utah and 45 (46%) attending health sciences graduate programs. Conclusions: FD student cohorts are more diverse than those in the University of Utah undergraduate and graduate programs, contributing to the diversity of those programs. More research is needed to ensure that graduate school gains are evidenced in all underrepresented groups.

Predator-Prey Interactions between Halobacteriovorax and Pathogenic Vibrio parahaemolyticus Strains: Geographical Considerations and Influence of Vibrio Hemolysins.

Halobacteriovorax is a genus of naturally occurring marine predatory bacteria that attack, replicate within, and lyse vibrios and other bacteria. This study evaluated the specificity of four Halobacteriovorax strains against important sequence types (STs) of clinically relevant Vibrio parahaemolyticus, including pandemic strains ST3 and ST36. The Halobacteriovorax bacteria were previously isolated from seawater from the Mid-Atlantic, Gulf of Mexico, and Hawaiian coasts of the United States. Specificity screening was performed using a double agar plaque assay technique on 23 well-characterized and genomically sequenced V. parahaemolyticus strains isolated from infected individuals from widely varying geographic locations within the United States. With few exceptions, results showed that Halobacteriovorax bacteria were excellent predators of the V. parahaemolyticus strains regardless of the origins of the predator or prey. Sequence types and serotypes of V. parahaemolyticus did not influence host specificity, nor did the presence or absence of genes for the thermostable direct hemolysin (TDH) or the TDH-related hemolysin, although faint (cloudy) plaques were present when one or both hemolysins were absent in three of the Vibrio strains. Plaque sizes varied depending on both the Halobacteriovorax and Vibrio strains evaluated, suggesting differences in Halobacteriovorax replication and/or growth rates. The very broad infectivity of Halobacteriovorax toward pathogenic strains of V. parahaemolyticus makes Halobacteriovorax a strong candidate for use in commercial processing applications to enhance the safety of seafoods. IMPORTANCE Vibrio parahaemolyticus is a formidable obstacle to seafood safety. Strains pathogenic to humans are numerous and difficult to control, especially within molluscan shellfish. The pandemic spread of ST3 and ST36 has caused considerable concern, but many other STs are also problematic. The present study demonstrates broad predatory activity of Halobacteriovorax strains obtained along U.S. coastal waters from the Mid-Atlantic, Gulf Coast, and Hawaii toward strains of pathogenic V. parahaemolyticus. This broad activity against clinically relevant V. parahaemolyticus strains suggests a role for Halobacteriovorax in mediating pathogenic V. parahaemolyticus levels in seafoods and their environment as well as the potential application of these predators in the development of new disinfection technologies to reduce pathogenic vibrios in molluscan shellfish and other seafoods.

Correction: Development of a Tailored Mobile Phone-Based Intervention to Facilitate Parent-Child Communication and Build Human Papillomavirus Vaccine Confidence: Formative Qualitative Study.

[This corrects the article DOI: 10.2196/43041.].

Development of a Tailored Mobile Phone-Based Intervention to Facilitate Parent-Child Communication and Build Human Papillomavirus Vaccine Confidence: Formative Qualitative Study.

BACKGROUND: Human papillomavirus (HPV) vaccine hesitancy is on the rise, and provider communication is a first-line strategy to address parental concerns. The use of the presumptive approach and motivational interviewing by providers may not be enough to influence parental decision-making owing to the providers' limited time, self-efficacy, and skills to implement these strategies. Interventions to enhance provider communication and build parental HPV vaccine confidence have been undertested. Delivering tailored patient education to parents via mobile phones before they visit the health care provider may address time constraints during clinic visits and positively affect vaccine uptake. OBJECTIVE: This study aimed to describe the development and evaluate the acceptability of a mobile phone-based, family-focused intervention guided by theory to address concerns of HPV vaccine-hesitant parents before the clinic visit, as well as explore intervention use to facilitate parent-child communication. METHODS: The health belief model and theory of reasoned action guided intervention content development. A multilevel stakeholder engagement process was used to iteratively develop the HPVVaxFacts intervention, including a community advisory board review, a review by an advisory panel comprising HPV vaccine-hesitant parents, a health communications expert review, semistructured qualitative interviews with HPV vaccine-hesitant parents (n=31) and providers (n=15), and a content expert review. Inductive thematic analysis was used to identify themes in the interview data. RESULTS: The qualitative interviews yielded 4 themes: overall views toward mobile device use for health information, acceptability of HPVVaxFacts, facilitators of HPVVaxFacts use, and barriers to HPVVaxFacts use. In parent interviews after reviewing HPVVaxFacts prototypes, almost all parents (29/31, 94%) stated they intended to have their child vaccinated. Most of the parents stated that they liked the added adolescents' corner to engage in optional parent-child communication (ie, choice to share and discuss information with their child; 27/31, 87%) and shared decision-making in some cases (8/31, 26%). After incorporating all input, the final intervention consisted of a 10-item survey to identify the top 3 concerns of parents, followed by tailored education that was mapped to each of the following concerns: evidential messages, images or graphics to enhance comprehension and address low literacy, links to credible websites, a provider video, suggested questions to ask their child's physician, and an optional adolescents' corner to educate the patient and support parent-child communication. CONCLUSIONS: The multilevel stakeholder-engaged process used to iteratively develop this novel intervention for HPV vaccine-hesitant families can be used as a model to develop future mobile health interventions. This intervention is currently being pilot-tested in preparation for a randomized controlled trial aiming to increase HPV vaccination among adolescent children of vaccine-hesitant parents in a clinic setting. Future research can adapt HPVVaxFacts for other vaccines and use in other settings (eg, health departments and pharmacies).

An Outbreak Investigation of Vibrio parahaemolyticus Infections in the United States Linked to Crabmeat Imported from Venezuela: 2018.

Vibrio parahaemolyticus is the leading cause of seafood-related foodborne illness globally. In 2018, the U.S. federal, state, and local public health and regulatory partners investigated a multistate outbreak of V. parahaemolyticus infections linked to crabmeat that resulted in 26 ill people and nine hospitalizations. State and U.S. Food and Drug Administration (FDA) laboratories recovered V. parahaemolyticus, Salmonella spp., and Listeria monocytogenes isolates from crabmeat samples collected from various points of distribution and conducted phylogenetic analyses of whole-genome sequencing data. Federal, state, and local partners conducted traceback investigations to determine the source of crabmeat. Multiple Venezuelan processors that supplied various brands of crabmeat were identified, but a sole firm was not confirmed as the source of the outbreak. Travel restrictions between the United States and Venezuela prevented FDA officials from conducting on-site inspections of cooked crabmeat processors. Based on investigation findings, partners developed public communications advising consumers not to eat crabmeat imported from Venezuela and placed potentially implicated firms on import alerts. While some challenges limited the scope of the investigation, epidemiologic, traceback, and laboratory evidence identified the contaminated food and country of origin, and contributed to public health and regulatory actions, preventing additional illnesses. This multistate outbreak illustrates the importance of adhering to appropriate food safety practices and regulations for imported seafood.

Levels of Vibrio parahaemolyticus in Pacific Oysters (Crassostrea gigas) From Washington State Following Ambient Exposure and Chilling.

Vibrio parahaemolyticus illnesses, often associated with the consumption of raw or undercooked oysters, are most common in summer months when higher temperatures increase V. parahaemolyticus levels in the environment. In Washington, post-harvest controls focus on the time from harvest to temperature-controlled storage to minimize V. parahaemolyticus illness risk. This study examined the effect of post-harvest ambient storage on V. parahaemolyticus levels in Pacific oysters. Additionally, the effects of cooling method, icing and/or refrigeration, on V. parahaemolyticus levels in oysters were evaluated. Five independent trials were conducted during July and August of 2015. For each trial, oysters were harvested from Totten Inlet and exposed to ambient conditions for 0 h (immediately cooled), 1 h, 5 h, or 9 h, and then either iced or refrigerated. Total and pathogenic (tdh+/trh+) V. parahaemolyticus levels were determined via MPN real-time PCR. Data from each trial were analyzed independently due to differences in initial V. parahaemolyticus levels. Total V. parahaemolyticus levels in oysters increased relative to control (0 h I) levels after the 1 h ambient exposure in only one trial, but pathogenic V. parahaemolyticus levels did not significantly increase after the 1 h exposure. Total and pathogenic V. parahaemolyticus levels increased by 0.8-1.9 log MPN/g in 5 h exposed oysters and by 1.0-2.9 log MPN/g in 9 h exposed oysters compared to levels in 0 h I samples. Mean maximum temperature of 5 h and 9 h exposed samples increased to ≈29°C compared to ≈21°C in 0 h and 1 h exposures, which likely contributed to observed increases in V. parahaemolyticus levels. Total and pathogenic V. parahaemolyticus levels increased more often in oysters cooled by refrigeration than by ice; this was most notable for the longer ambient exposure samples. Overall, these data support shorter post-harvest ambient exposure as well as rapid cooling of oysters to minimize risk of V. parahaemolyticus illness.

Project ECHO for Cancer Care: a Scoping Review of Provider Outcome Evaluations.

The Project ECHO model of telementoring has been used for the past 10 years to expand access to specialized cancer care. This scoping review identifies evidence for the model's ability to improve provider outcomes, synthesizing findings from existing studies within Moore et al.'s (2009) framework for continuing medical education outcomes. We search two large research databases and a collection maintained by Project ECHO staff for articles that focus on cancer ECHO programs, involve primary data collection, and were published between December 1, 2016, and November 30, 2021. We identified 25 articles for inclusion in our scoping review. Most articles reported results for outcomes related to program participation: attendance, satisfaction, and learning. Yet, just under half reported changes in provider practices. Results demonstrate widespread participation and improved learning resulting from ECHO programs focused on cancer care. There is also evidence of improved practices related to HCV vaccination and palliative care. We highlight examples of best practices as well as opportunities to improve provider outcome evaluations for cancer ECHO programs.

Environmental Drivers of Vibrio cholerae Abundances in Mobile Bay, Alabama.

Vibrio cholerae is the etiological agent of the illness cholera. However, there are non-O1/non-O139 V. cholerae (NOVC) strains that generally lack the toxin gene (ctx) and colonization factors that cause cholera. These NOVC strains are autochthonous members of estuarine environments and a significant cause of seafood-borne gastroenteritis in the United States. The objective of this study was to identify environmental parameters that correlate with NOVC prevalence in oysters, water, and sediment at three ecologically diverse locations in Mobile Bay, AL, including Dog River (DR), Fowl River (FR), and Cedar Point (CP). Oyster, water, and sediment samples were collected twice a month when conditions were favorable for NOVC growth and once a month when they were not. A most probable number (MPN)/real-time PCR assay was used to determine NOVC abundances. Environmental parameters were measured during sampling to determine their relationship, if any, with NOVC at each site. NOVC abundances in oysters at DR, FR, and CP were 0.87, 0.87, and -0.13 log MPN/g, respectively. In water, the median NOVC levels at DR, FR, and CP were 1.18, -0.13, and -0.82 log MPN/mL, and in sediment, the levels were 1.48, 1.87, and -0.03 log MPN/g, respectively. Correlations of NOVC abundances in oyster, water, and sediment samples with environmental parameters were largely site specific. For example, the levels of NOVC in oysters at DR had a positive correlation with temperature but a negative correlation with dissolved oxygen (DO) and nutrient concentrations, NO2-, NO3-, dissolved inorganic nitrogen (DIN), total dissolved nitrogen (TDN), and dissolved inorganic phosphorus (DIP). At FR, however, the levels of NOVC in oysters displayed only a negative correlation with NO2-. When grouping NOVC abundances by temperature, the main driving factor for prevalence, additional correlations with salinity, total cell counts, dissolved organic nitrogen (DON), and dissolved organic carbon (DOC) became evident regardless of the site. IMPORTANCE NOVC can cause gastrointestinal illness in humans, which typically occurs after the consumption of raw or undercooked seafood. Incidence rates of NOVC gastroenteritis have increased during the past decade. In this study, NOVC was enumerated from oysters, sediment, and water collected at three sites in Mobile Bay, with environmental parameters measured concurrently over the course of a year, to identify potential environmental drivers of NOVC abundances. The data from this study, from an area lacking in V. cholerae research, provide a useful baseline for risk analysis of V. cholerae infections. Defining correlations between NOVC and environmental attributes at different sites and temperatures within a dynamic system such as Mobile Bay provides valuable data to better understand the occurrence and proliferation of V. cholerae in the environment.

Teachers as Youth Suicide Prevention Gatekeepers: An Examination of Suicide Prevention Training and Exposure to Students at Risk of Suicide.

Background: Teachers are important gatekeepers in suicide prevention for children and youth, yet little is known about factors that contribute to suicide prevention training effectiveness and the influence of student suicidality on teachers' role as gatekeepers. Objective: This study examined teachers' attitudes and self-efficacy in suicide prevention including an examination of suicide prevention training and exposure to student suicidality. Researchers examined incremental prediction of the relationship between teachers' self-efficacy, outcome expectations, and outcome values following prevention training. Methods: Participants included teachers in PreK-12th grade schools in the United States (N = 505). Researchers used non-parametric statistics to examine group level differences and a structural equation model (SEM) to test the proposed theoretical model. Results: Teachers who experienced a student death by suicide reported significantly higher levels of gatekeeper reluctance than teachers who had not experienced a student death by suicide (p < 0.01). Similarly, teachers who encountered students with suicidal thoughts reported greater levels of gatekeeper reluctance (p < 0.01) and higher self-efficacy to engage in suicide prevention (p < 0.05) compared to teachers who had not had this exposure. Results of the SEM indicated an adequate goodness of fit and fit statistics [χ2 (87) = 194.420, p = 0.000; CFI = 0.95; RMSEA = 0.05]. The model remained in-tact when exposure to student suicide was added. Conclusions: Findings support the importance of supporting teachers continued engagement in youth suicide prevention and prevention training that targets specific outcomes in teachers' attitudes and efficacy. Supplementary Information: The online version contains supplementary material available at 10.1007/s10566-022-09699-5.

School and electronic bullying among adolescents: Direct and indirect relationships with sadness, sleep, and suicide ideation.

INTRODUCTION: Bullying is an increasing concern for education, health, and policy. Adolescence is a particularly vulnerable period for the development of depressive symptoms and suicidality following exposure to bullying. However, limited research investigating the potential impact of depressive symptoms on the bullying-suicide relationship exists. METHODS: Using national data (N = 13,677) from the most recent 2019 Youth Risk Behavior Survey, the aim of the present study was to examine the prevalence among adolescents' (school/electronic) bullying victimization, depressive symptoms (sadness; sleep), and suicide ideation as well as their associations including direct and indirect relationships including exploring differences by gender and race/ethnicity. RESULTS: Descriptive results indicated an increase in the prevalence of adolescents being bullied (both on school property and electronically), experiencing feelings of sadness, and hopelessness as well as a decrease in getting more than 8 h of sleep between 2017 and 2019. In 2019, over one-third of respondents felt sad or hopeless almost every day for 2 weeks or more in a row, which stopped them from doing some usual activities. Structural equation modeling indicated that (school/electronic) bullying was directly associated with feelings of sadness/hopelessness and suicide ideation, with sadness mediating the link between bullying and suicide ideation. CONCLUSION: Now more than ever, it is critical to promote the collaboration between educators, mental health specialists, policymakers, and researchers to develop and implement evidence-based strategies and approaches to preventing and reducing both bullying victimization and the associated psychological distress and mental health outcomes.

Increasing URiM Family Medicine Residents at University of Utah Health.

INTRODUCTION: The Department of Family and Preventive Medicine is home for the University of Utah's Family Medicine Residency program. Although Utah's diversity is steadily increasing, the race/ethnic diversity of the program's family medicine residency does not reflect the state's general population. METHODS: From 2017 to 2021, the residency instituted several adjustments to recruitment processes, including modification of an existing screening system to better highlight resiliency in overcoming challenging life experiences; promotion of commitment to diversity during interview days; incorporation of increased participation from diverse faculty and residents on interview days; and addition of outreach from the Office of Health, Equity, Diversity, and Inclusion. Underrepresented in medicine (URiM) applicants were the first to be offered interviews in an identical screening score cohort, and were ranked highest in rank lists in cohorts with identical final rank scores. RESULTS: Over the past five match cycles, Latinx residents have increased from zero to six, and underrepresented Asian residents from zero to two. In the 2021 match cycle, five of 10 incoming residents (50%) are URiM. Overall, URiM residents are now 30%, and residents of color 36%, of a total of 30 residents across all 3 training years. We found that eight URiM interviews were needed for every one URiM match. CONCLUSION: Intentional resident recruitment initiatives can transform racial/ethnic diversity in a family medicine residency program in a short amount of time.

Effects of Farm Location on Vibrio parahaemolyticus and Vibrio vulnificus Levels in Oysters after Desiccation and Resubmersion in the Northern Gulf of Mexico.

ABSTRACT: Desiccation is a routine farming practice used in off-bottom oyster aquaculture to reduce biofouling organisms and improve shell quality. This practice can increase Vibrio parahaemolyticus and Vibrio vulnificus levels, leading to increased risk of illness for raw oyster consumers. Previous resubmersion studies were performed in geographic proximity to one another, so to better understand the broader applicability of resubmersion, the next step was to perform concurrent studies in multiple geographic locations within a region. This study evaluated the effect of variations in geographic location on the recovery time needed for elevated vibrio levels to return to ambient levels in desiccated oysters after resubmersion at Gulf Coast farms. Two trials were performed between May and August 2019 at sites spanning ∼100 km: three in Alabama and one in Florida. Oysters were deployed in OysterGro cages at each location, 2 weeks before each trial, and then either were desiccated for 24 h or remained submersed as controls. Triplicate samples were taken before and immediately following the desiccation period, as well as 7 and 14 days after resubmersion. Total and pathogenic V. parahaemolyticus and V. vulnificus levels were determined using most-probable-number (MPN) real-time PCR. Vibrio levels increased by 0.23 to 3.50 log MPN/g after desiccation. Recovery times varied among geographic locations by trial and Vibrio spp., with all vibrio counts recovering to levels not significantly higher than those in control oysters within 7 to 14 days of resubmersion (P ≥ 0.06). These results suggest a 14-day resubmersion period of cultured oysters allowed vibrio levels, elevated because of routine handling, to return to ambient levels at all farm sites studied.

Biochemical and Virulence Characterization of Vibrio vulnificus Isolates From Clinical and Environmental Sources.

Vibrio vulnificus is a deadly human pathogen for which infections occur via seafood consumption (foodborne) or direct contact with wounds. Virulence is not fully characterized for this organism; however, there is evidence of biochemical and genotypic correlations with virulence potential. In this study, biochemical profiles and virulence genotype, based on 16S rRNA gene (rrn) and virulence correlated gene (vcg) types, were determined for 30 clinical and 39 oyster isolates. Oyster isolates were more biochemically diverse than the clinical isolates, with four of the 20 tests producing variable (defined as 20-80% of isolates) results. Whereas, for clinical isolates only mannitol fermentation, which has previously been associated with virulence potential, varied among the isolates. Nearly half (43%) of clinical isolates were the more virulent genotype (rrnB/vcgC); this trend was consistent when only looking at clinical isolates from blood. The majority (64%) of oyster isolates were the less virulent genotype (rrnA or AB/vcgE). These data were used to select a sub-set of 27 isolates for virulence testing with a subcutaneously inoculated, iron-dextran treated mouse model. Based on the mouse model data, 11 isolates were non-lethal, whereas 16 isolates were lethal, indicating a potential for human infection. Within the non-lethal group there were eight oyster and three clinical isolates. Six of the non-lethal isolates were the less virulent genotype (rrnA/vcgE or rrnAB/vcgE) and two were rrnB/vcgC with the remaining two of mixed genotype (rrnAB/vcgC and rrnB/vcgE). Of the lethal isolates, five were oysters and 11 were clinical. Eight of the lethal isolates were the less virulent genotype and seven the more virulent genotype, with the remaining isolate a mixed genotype (rrnA/vcgC). A discordance between virulence genotype and individual mouse virulence parameters (liver infection, skin infection, skin lesion score, and body temperature) was observed; the variable most strongly associated with mouse virulence parameters was season (warm or cold conditions at time of strain isolation), with more virulent strains isolated from cold conditions. These results indicate that biochemical profiles and genotype are not significantly associated with virulence potential, as determined by a mouse model. However, a relationship with virulence potential and seasonality was observed.

Phylogenetic and Biogeographic Patterns of Vibrio parahaemolyticus Strains from North America Inferred from Whole-Genome Sequence Data.

Vibrio parahaemolyticus is the most common cause of seafood-borne illness reported in the United States. The draft genomes of 132 North American clinical and oyster V. parahaemolyticus isolates were sequenced to investigate their phylogenetic and biogeographic relationships. The majority of oyster isolate sequence types (STs) were from a single harvest location; however, four were identified from multiple locations. There was population structure along the Gulf and Atlantic Coasts of North America, with what seemed to be a hub of genetic variability along the Gulf Coast, with some of the same STs occurring along the Atlantic Coast and one shared between the coastal waters of the Gulf and those of Washington State. Phylogenetic analyses found nine well-supported clades. Two clades were composed of isolates from both clinical and oyster sources. Four were composed of isolates entirely from clinical sources, and three were entirely from oyster sources. Each single-source clade consisted of one ST. Some human isolates lack tdh, trh, and some type III secretion system (T3SS) genes, which are established virulence genes of V. parahaemolyticus Thus, these genes are not essential for pathogenicity. However, isolates in the monophyletic groups from clinical sources were enriched in several categories of genes compared to those from monophyletic groups of oyster isolates. These functional categories include cell signaling, transport, and metabolism. The identification of genes in these functional categories provides a basis for future in-depth pathogenicity investigations of V. parahaemolyticusIMPORTANCEVibrio parahaemolyticus is the most common cause of seafood-borne illness reported in the United States and is frequently associated with shellfish consumption. This study contributes to our knowledge of the biogeography and functional genomics of this species around North America. STs shared between the Gulf Coast and the Atlantic seaboard as well as Pacific waters suggest possible transport via oceanic currents or large shipping vessels. STs frequently isolated from humans but rarely, if ever, isolated from the environment are likely more competitive in the human gut than other STs. This could be due to additional functional capabilities in areas such as cell signaling, transport, and metabolism, which may give these isolates an advantage in novel nutrient-replete environments such as the human gut.

The Teacher Expectations and Values for Suicide Prevention Scale.

Background: Teachers play a critical role in youth suicide prevention, yet few psychometrically sound instruments exist to measure teachers' expectations and values regarding suicide prevention. Aims: This study examined the factor structure and psychometric properties of the Teacher Expectations and Values for Suicide Prevention (TEVSP) Scale. Method: The TEVSP was administered to 500 teachers in the United States. Confirmatory factor analysis was used to determine the factor structure and bivariate correlations were used to investigate convergent and discriminant validity. Mann-Whitney U tests investigated group differences in TEVSP scores between participants who had received suicide training and those who had not. Results: Results support a three-factor hierarchical model with strong internal consistency and evidence of validity. Significant differences were found in TEVSP scores between groups. Limitations: There is a need to further explore the psychometric properties of the scale across samples and face-to-face methods. Conclusion: The TEVSP is a sound instrument that can be used to measure teachers' expectations and values for suicide prevention.

Effect of Gear Type on Vibrio spp. Levels in Farm-Raised Oysters (Crassostrea virginica) after Routine Handling and Resubmersion.

ABSTRACT: During routine handling, cultured oysters are removed from the water and exposed to elevated temperatures, causing growth of Vibrio vulnificus and Vibrio parahaemolyticus within them. Farmers can resubmerse oysters in the water, allowing elevated Vibrio spp. levels to return to ambient levels within the oysters. Previous resubmersion research is limited to one aquaculture gear type during studies performed from June to September. This study aims to expand knowledge about the recovery times needed for elevated Vibrio levels in handled oysters from two common gear types (the adjustable longline system and the OysterGro system) during early and midsummer periods. Oysters held in both gear types were subjected to being tumbled and refrigerated or desiccated and then resubmersed into water in May and July 2018 and 2019. Vibrio spp. levels were measured before and after the treatments, and 3, 7, and 14 days after resubmersion, and were compared with levels in submersed oysters. All samples were tested for V. vulnificus, total V. parahaemolyticus, and pathogenic V. parahaemolyticus (tdh+/trh+). Water temperatures in May were significantly lower (∼5°C; P ≤ 0.009) than in July, corresponding to lower V. vulnificus levels (-0.67 log MPN/g) and higher tdh+/trh+ levels (+0.56 to 0.63 log MPN/g) in control oysters. The average Vibrio spp. levels in control oysters from each trial did not differ between the gear types (P ≥ 0.08). Elevated V. vulnificus levels recovered to ambient levels after 7 days in May and 3 days in July, regardless of gear or handling. For V. parahaemolyticus, the desiccated oysters required 14 days to recover in May and 7 days in July, whereas the tumbled and refrigerated oysters required 14 days or more in both months. This study had limited replication in each month, but the data suggest that the resubmersion times differ between the gear types, treatment types, and months. Future studies with more replications are needed to determine whether these trends continue.

Effects of tumbling, refrigeration and subsequent resubmersion on the abundance of Vibrio vulnificus and Vibrio parahaemolyticus in cultured oysters (Crassostrea virginica).

Routine handling of oysters is a common industry practice for off-bottom oyster aquaculture, which aims to produce a high-quality oyster. These practices expose oysters to elevated temperatures and interrupt filter feeding, which can increase Vibrio vulnificus and V. parahaemolyticus levels within the oyster. The resubmersion of oysters after exposure to conditions where the time-temperature controls are exceeded is as an effective mitigation strategy to allow elevated levels of Vibrio spp. to "recover", or return to ambient levels, prior to harvest. Previous work examined the effect of desiccation on recovery times; the objective of this study was to evaluate the effect of additional handling treatments [tumbled and refrigerated (TR), tumbled and not refrigerated (TNR), not tumbled and refrigerated (NTR), and not tumbled and not refrigerated (NTNR)] on the time needed for V. vulnificus, total V. parahaemolyticus, and pathogenic V. parahaemolyticus (tdh+/trh+) to recover in oysters. A set of non-treated (control) oysters remained submerged throughout the study to determine the ambient Vibrio spp. (inclusive of genotypes) levels within oysters. Vibrio spp. levels were measured immediately before (pre) and after (post) the treatments, and 1, 2, 4, 7, 10, and 14 days after resubmersion using a three-tube MPN real-time PCR method. The non-refrigerated oysters (TNR, NTNR) had Vibrio spp. levels 1.54 to 2.10 log MPN/g higher than the pre-treatment levels, while the Vibrio spp. levels in refrigerated oysters were not significantly higher than pre-treatment levels. After resubmersion, Vibrio spp. levels increased by 0.84 to 1.78 log MPN/g in the refrigerated oysters (TR, NTR). Vibrio spp. levels in oysters returned to ambient after 1-7 days of resubmersion, depending on the handling treatment and the Vibrio spp. These results provide data on handling treatments not previously reported and further support the seven-day resubmersion requirement for farmers in Alabama using the adjustable longline system.