Co-infection is linked to infection prevalence and intensity in oysters amidst high environmental and spatial variation.

Co-infecting parasites modify infection outcomes in the wild. However, it is unclear how multiple environmental factors influence co-infection. The Chesapeake Bay metapopulation of the eastern oyster, Crassostrea virginica, provides an opportunity to test the importance of co-infection across heterogeneous environments because multiple parasites infect oysters across a broad salinity gradient. This study leverages Maryland and Virginia oyster monitoring for a large-scale survey of four co-infecting organisms, including two tissue parasites and two shell bio-eroding parasites. We diagnosed infection in 440 oysters across 16 paired harvested and unharvested reefs and tested the importance of co-infecting organisms for each parasite relative to environmental conditions, host traits, and marine spatial management. Microscopic visual methods were used to diagnose prevalence and intensity of tissue infections with Perkinsus marinus (the causative agent of dermo disease) and Haplosporidium nelsoni (the causative agent of MSX disease). Macroscopic visual methods were used to diagnose prevalence and intensity of shell infections with Cliona boring sponges and blister-inducing Polydora worms. For the three oyster parasites that were detected [H. nelsoni infections were absent in all oysters], salinity was the overall strongest predictor, corresponding to bay-wide patterns of parasite prevalence and/or intensity. Despite high environmental and spatial variation, co-infections corresponded to altered prevalence and/or intensity for all three oyster parasites. The correlational patterns suggest that P. marinus acts as a lynchpin in co-infection, as its intensity increased with Cliona sponge prevalence and P. marinus co-infection predicted higher Polydora blister intensity. Oyster shell height, reef habitat, and harvest status also predicted parasite prevalence and intensity, further reflecting the multivariate drivers of infections in this system. Unharvested reefs had greater vertical habitat structure and higher intensities of Cliona sponge infections, but no differences in the prevalence of any of the three parasites. Spatial patterns unexpectedly show that reef-level predictors of parasite patterns were more important than differences between tributaries. This correlational survey provides novel insights through the statistical relationships between the three oyster parasites, environmental conditions, host traits, and human resource management. New and more detailed scenarios are needed to expand disease ecological theory to encompass co-infection in anthropogenically impacted wildlife populations.

High School Follow-Up of the Dating Matters® RCT: Effects on Teen Dating Violence and Relationship Behaviors.

Teen dating violence (TDV) is a significant public health problem that can have lifelong consequences. Using a longitudinal, cluster randomized controlled trial (RCT), this study examines whether the Dating Matters comprehensive prevention model, implemented in middle school, prevented TDV and negative relationship behaviors and promoted positive relationship behaviors in high school (9th-11th grades), when compared with a standard of care intervention. Dating Matters includes programs for sixth to eighth grade youth and their parents, training for school staff, a youth communications program, and policy and data activities implemented in the community. Self-report survey data were collected from students in 46 middle schools that were randomly assigned to condition within site. Students completed two surveys (fall and spring) in each middle school grade and a single survey in the spring of each high school grade. This study examined self-reported TDV perpetration and victimization, use of negative conflict resolution strategies, and positive relationship skills in the high school follow-up. While varying patterns emerged, latent panel models demonstrated significant program effects for all outcomes. Dating Matters students reported 19% reduced risk for TDV perpetration, 24% reduced risk for TDV victimization, 7% reduced risk for use of negative conflict strategies, and 3% more use of positive relationship skills, on average across time and cohort, than standard of care students. On average, Dating Matters, implemented in middle school, continued to be more effective at reducing TDV perpetration, TDV victimization, and use of negative conflict resolution strategies in high school than an evidence-based comparison program.Trial Registration: clinicaltrials.gov Identifier: NCT01672541.

Patterns of adverse childhood experiences and their associations with mental distress, substance use and sexual risk behaviors in Sub-Saharan Africa.

BACKGROUND: Multiple adverse childhood experiences (ACEs) are associated with poor sexual and mental health outcomes in low- and middle-income countries (LMICs). Less well understood is how discrete and gendered clustering of ACEs may influence health. OBJECTIVE: To assess how multiple ACEs co-occur and how dominant patterns of co-occurrence are associated with mental distress, substance use, and sexual risk behaviors among young women and men in Sub-Saharan Africa. PARTICIPANTS AND SETTING: We used pooled data of young men and women aged 19-24 from comparable, nationally representative Violence Against Children and Youth Surveys (VACS) conducted in Cote d'Ivoire, Kenya, Lesotho, Mozambique, and Namibia (nf = 7183; nm = 2207). METHODS: We estimated sex-disaggregated latent classes of six ACEs among young women and men. We ran Bolck-Croon-Hagenaars (BCH) distal outcome analysis to test the sex-stratified relationships between ACEs latent classes and health outcomes. RESULTS: A six class solution best fit the female data. Classes included witnessing violence and experiencing physical violence (PV); experiencing PV; high ACEs; witnessing community violence; orphanhood; and low ACEs exposure. Among males, the best-fitting three-class solution included experiencing PV and witnessing community violence; high ACEs; and low ACEs exposure. Membership in the high ACEs class was associated with mental distress among females and males, and substance use among males. No differences in sexual risk behavior were identified by class membership among either females or males. CONCLUSIONS: Discrete clusters of co-occurring ACEs are associated with elevated odds of mental distress among females, and mental distress and substance use among males. Preventing ACEs may improve mental health among young women and men in LMICs in Sub-Saharan Africa.

Prevalence of Violence Victimization and Perpetration During Middle and High School in Underresourced, Urban Communities.

This study describes rates of violence victimization, perpetration, and witnessing in 6th-11th grade for a multisite sample (N = 3,466) of predominantly Black and Hispanic middle- and high-school students from urban areas with high rates of crime and economic disadvantage. Students completed surveys in middle and high school assessing teen dating violence, stalking, sexual violence and harassment, bullying, cyberbullying, and physical violence perpetration and victimization, as well as witnessing violence. The highest prevalence rates are observed most often in 8th or 9th grade. Youth reported high rates of witnessing serious assault and severe community violence throughout adolescence. These findings suggest that efforts to prevent violence among youth living in under-resourced communities need to start early and address community-level socioeconomic disparities.

Middle School Effects of the Dating Matters® Comprehensive Teen Dating Violence Prevention Model on Physical Violence, Bullying, and Cyberbullying: a Cluster-Randomized Controlled Trial.

Few comprehensive primary prevention approaches for youth have been evaluated for effects on multiple types of violence. Dating Matters®: Strategies to Promote Healthy Teen Relationships (Dating Matters) is a comprehensive teen dating violence (TDV) prevention model designed by the Centers for Disease Control and Prevention and evaluated using a longitudinal stratified cluster-randomized controlled trial to determine effectiveness for preventing TDV and promoting healthy relationship behaviors among middle school students. In this study, we examine the prevention effects on secondary outcomes, including victimization and perpetration of physical violence, bullying, and cyberbullying. This study examined the effectiveness of Dating Matters compared to a standard-of-care TDV prevention program in 46 middle schools in four high-risk urban communities across the USA. The analytic sample (N = 3301; 53% female; 50% Black, non-Hispanic; and 31% Hispanic) consisted of 6th-8th grade students who had an opportunity for exposure to Dating Matters in all three grades or the standard-of-care in 8th grade only. Results demonstrated that both male and female students attending schools implementing Dating Matters reported 11% less bullying perpetration and 11% less physical violence perpetration than students in comparison schools. Female Dating Matters students reported 9% less cyberbullying victimization and 10% less cyberbullying perpetration relative to the standard-of-care. When compared to an existing evidence-based intervention for TDV, Dating Matters demonstrated protective effects on physical violence, bullying, and cyberbullying for most groups of students. The Dating Matters comprehensive prevention model holds promise for reducing multiple forms of violence among middle school-aged youth. ClinicalTrials.gov Identifier: NCT01672541.

Effects of the Dating Matters® Comprehensive Prevention Model on Health- and Delinquency-Related Risk Behaviors in Middle School Youth: a Cluster-Randomized Controlled Trial.

Teen dating violence (TDV) is associated with a variety of delinquent behaviors, such as theft, and health- and delinquency-related risk behaviors, including alcohol use, substance abuse, and weapon carrying. These behaviors may co-occur due to shared risk factors. Thus, comprehensive TDV-focused prevention programs may also impact these other risk behaviors. This study examined the effectiveness of CDC's Dating Matters®: Strategies to Promote Healthy Teen Relationships (Dating Matters) comprehensive TDV prevention model compared to a standard-of-care condition on health- and delinquency-related risk behaviors among middle school students. Students (N = 3301; 53% female; 50% black, non-Hispanic; and 31% Hispanic) in 46 middle schools in four sites across the USA were surveyed twice yearly in 6th, 7th, and 8th grades. A structural equation modeling framework with multiple imputation to account for missing data was utilized. On average over time, students receiving Dating Matters scored 9% lower on a measure of weapon carrying, 9% lower on a measure of alcohol and substance abuse, and 8% lower on a measure of delinquency by the end of middle school than students receiving an evidence-based standard-of-care TDV prevention program. Dating Matters demonstrated protective effects for most groups of students through the end of middle school. These results suggest that this comprehensive model is successful at preventing risk behaviors associated with TDV. clinicaltrials.gov Identifier: NCT01672541.

Effects of Dating Matters® on Sexual Violence and Sexual Harassment Outcomes among Middle School Youth: a Cluster-Randomized Controlled Trial.

Sexual violence (SV), including sexual harassment (SH), is a significant public health problem affecting adolescent health and well-being. This study extends prior research by evaluating the effectiveness of a comprehensive teen dating violence prevention model, Dating Matters, on SV and SH perpetration and victimization, inclusive of any victim-perpetrator relationship, among middle school students. Dating Matters includes classroom-delivered programs for youth in 6th, 7th, and 8th grades; community-based programs for parents; a youth communications program; training for educators; and community-level activities. Middle schools in four urban areas in the USA were randomly assigned to receive Dating Matters (DM, N = 22) or a standard-of-care intervention (SC, N = 24) over four consecutive school years (2012-2016). The analytic sample included two cohorts who entered the study in 6th grade and completed 8th grade by the end of the study allowing for full exposure to Dating Matters (DM: N = 1662; SC: N = 1639; 53% female; 50% black, non-Hispanic; 6 waves of data collection for each cohort). Structural equation modeling was employed with multiple imputation to account for missing data. Dating Matters was associated with significant reductions in SV and SH perpetration and victimization scores in most-but not all-sex/cohort groups by the end of 8th grade relative to an evidence-based TDV prevention program. On average, students receiving Dating Matters scored 6% lower on SV perpetration, 3% lower on SV victimization, 4% lower on SH perpetration, and 8% lower on SH victimization by the end of middle school than students receiving an evidence-based violence prevention program. Overall, Dating Matters shows promise for reducing SV and SH, occurring both within and outside dating relationships, through middle school. Clinicaltrials.gov Identifier: NCT01672541.

Ecological Factors Mediate Immunity and Parasitic Co-Infection in Sea Fan Octocorals.

The interplay among environment, demography, and host-parasite interactions is a challenging frontier. In the ocean, fundamental changes are occurring due to anthropogenic pressures, including increased disease outbreaks on coral reefs. These outbreaks include multiple parasites, calling into question how host immunity functions in this complex milieu. Our work investigates the interplay of factors influencing co-infection in the Caribbean sea fan octocoral, Gorgonia ventalina, using metrics of the innate immune response: cellular immunity and expression of candidate immune genes. We used existing copepod infections and live pathogen inoculation with the Aspergillus sydowii fungus, detecting increased expression of the immune recognition gene Tachylectin 5A (T5A) in response to both parasites. Cellular immunity increased by 8.16% in copepod infections compared to controls and single Aspergillus infections. We also detected activation of cellular immunity in reef populations, with a 13.6% increase during copepod infections. Cellular immunity was similar in the field and in the lab, increasing with copepod infections and not the fungus. Amoebocyte density and the expression of T5A and a matrix metalloproteinase (MMP) gene were also positively correlated across all treatments and colonies, irrespective of parasitic infection. We then assessed the scaling of immune metrics to population-level disease patterns and found random co-occurrence of copepods and fungus across 15 reefs in Puerto Rico. The results suggest immune activation by parasites may not alter parasite co-occurrence if factors other than immunity prevail in structuring parasite infection. We assessed non-immune factors in the field and found that sea fan colony size predicted infection by the copepod parasite. Moreover, the effect of infection on immunity was small relative to that of site differences and live coral cover, and similar to the effect of reproductive status. While additional immune data would shed light on the extent of this pattern, ecological factors may play a larger role than immunity in controlling parasite patterns in the wild. Parsing the effects of immunity and ecological factors in octocoral co-infection shows how disease depends on more than one host and one parasite and explores the application of co-infection research to a colonial marine organism.

Warming and pollutants interact to modulate octocoral immunity and shape disease outcomes.

Warming environments can alter the outcome of host-parasite relationships with important consequences for biodiversity. Warming often increases disease risk, and interactions with other environmental factors can intensify impacts by modifying the underlying mechanisms, such as host immunity. In coastal ecosystems, metal pollution is a pervasive stressor that influences disease and immunity in many organisms. Despite the crisis facing coral reefs, which stems in part from warming-associated disease outbreaks, the impacts of metal pollutants on scleractinian and octocoral disease are largely unknown. We investigated how warming oceans and copper pollution affect host immunity and disease risk for two diseases of the abundant Caribbean octocoral, the sea fan Gorgonia ventalina. Field surveys across a sediment copper concentration gradient in Puerto Rico, USA revealed that cellular immunity of sea fans increased by 12.6% at higher sediment copper concentrations, while recovery from multifocal purple spots disease (MFPS) tended to decrease. MFPS severity in the field increased at warmer sites. In a controlled laboratory experiment, sea fans were inoculated with live cultures of a labyrinthulid parasite to test the interactive effects of temperature and copper on immune activation. As in the field, higher copper induced greater immunity, but the factorial design of the experiment revealed that copper and temperature interacted to modulate the immune response to the parasite: immune cell densities increased with elevated temperature at lower copper concentrations, but not with high copper concentrations. Tissue damage was also greater in treatments with higher copper and warmer temperatures. Field and lab evidence confirm that elevated copper hinders sea fan immune defenses against damaging parasites. Temperature and copper influenced host-pathogen interactions in octocorals by modulating immunity, disease severity, and disease recovery. This is the first evidence that metal pollution affects processes influencing disease in octocorals and highlights the importance of immune mechanisms in environmentally mediated disease outbreaks. Although coral conservation efforts must include a focus on global factors, such as rapid warming, reducing copper and other pollutants that compromise coral health on a local scale may help corals fight disease in a warming ocean.

Increases and decreases in marine disease reports in an era of global change.

Outbreaks of marine infectious diseases have caused widespread mass mortalities, but the lack of baseline data has precluded evaluating whether disease is increasing or decreasing in the ocean. We use an established literature proxy method from Ward and Lafferty (Ward and Lafferty 2004 PLoS Biology2, e120 (doi:10.1371/journal.pbio.0020120)) to analyse a 44-year global record of normalized disease reports from 1970 to 2013. Major marine hosts are combined into nine taxonomic groups, from seagrasses to marine mammals, to assess disease swings, defined as positive or negative multi-decadal shifts in disease reports across related hosts. Normalized disease reports increased significantly between 1970 and 2013 in corals and urchins, indicating positive disease swings in these environmentally sensitive ectotherms. Coral disease reports in the Caribbean correlated with increasing temperature anomalies, supporting the hypothesis that warming oceans drive infectious coral diseases. Meanwhile, disease risk may also decrease in a changing ocean. Disease reports decreased significantly in fishes and elasmobranchs, which have experienced steep human-induced population declines and diminishing population density that, while concerning, may reduce disease. The increases and decreases in disease reports across the 44-year record transcend short-term fluctuations and regional variation. Our results show that long-term changes in disease reports coincide with recent decades of widespread environmental change in the ocean.

An RCT of Dating Matters: Effects on Teen Dating Violence and Relationship Behaviors.

INTRODUCTION: Teen dating violence is a serious public health problem with few effective prevention strategies. This study examines whether the Dating Matters comprehensive prevention model, compared with a standard of care intervention, prevented negative relationship behaviors and promoted positive relationship behaviors. STUDY DESIGN: This longitudinal, cluster-RCT compared the effectiveness of Dating Matters with standard of care across middle school. Standard of care was an evidence-based teen dating violence prevention curriculum (Safe Dates) implemented in eighth grade. SETTING/PARTICIPANTS: Forty-six middle schools in high-risk urban neighborhoods in four U.S. cities were randomized. Schools lost to follow-up were replaced with new schools, which were independently randomized (71% school retention). Students were surveyed in fall and spring of sixth, seventh, and eighth grades (2012-2016). The analysis sample includes students from schools implementing Dating Matters or standard of care for >2 years who started sixth grade in the fall of 2012 or 2013 and had dated (N=2,349 students, mean age 12 years, 49% female, and 55% black, non-Hispanic, 28% Hispanic, 17% other). INTERVENTION: Dating Matters is a comprehensive, multicomponent prevention model including classroom-delivered programs for sixth to eighth graders, training for parents of sixth to eighth graders, educator training, a youth communications program, and local health department activities to assess capacity and track teen dating violence-related policy and data. MAIN OUTCOME MEASURES: Self-reported teen dating violence perpetration and victimization, use of negative conflict resolution strategies, and positive relationship skills were examined as outcomes. Imputation and analyses were conducted in 2017. RESULTS: Latent panel models demonstrated significant program effects for three of four outcomes; Dating Matters students reported 8.43% lower teen dating violence perpetration, 9.78% lower teen dating violence victimization, and 5.52% lower use of negative conflict resolution strategies, on average across time points and cohorts, than standard of care students. There were no significant effects on positive relationship behaviors. CONCLUSIONS: Dating Matters demonstrates comparative effectiveness, through middle school, for reducing unhealthy relationship behaviors, such as teen dating violence and use of negative conflict resolution strategies, relative to the standard of care intervention. TRIAL REGISTRATION: This study is registered at www.clinicaltrials.gov NCT01672541.

Octocoral co-infection as a balance between host immunity and host environment.

Co-infection is the reality in natural populations, but few studies incorporate the players that matter in the wild. We integrate the environment, host demography, two parasites, and host immunity in a study of co-infection to determine the drivers of parasite interactions. Here, we use an ecologically important Caribbean sea fan octocoral, Gorgonia ventalina, that is co-infected by a copepod and a labyrinthulid protist. We first expanded upon laboratory studies by showing that immune suppression is associated with the labyrinthulid in a natural setting. Histological analyses revealed that immune cells (amoebocytes) were significantly suppressed in both labyrinthulid infections and co-infections relative to healthy sea fans, but remained unchanged in copepod infections. However, surveys of natural coral populations demonstrated a critical role for the environment and host demography in this co-infection: the prevalence of copepod infections increased with sea fan size while labyrinthulid prevalence increased with water depth. Although we predicted that immune suppression by the labyrinthulid would facilitate copepod infection, the two parasites did not co-occur in the sea fans more often than expected by chance. These results suggest that the distinct ecological drivers for each parasite overwhelm the role of host immune suppression in determining the distribution of parasites among hosts. This interplay of the environment and parasite-mediated immune suppression in sea fan co-infection provides insights into the factors underlying co-occurrence patterns in wild co-infections. Moving forward, simultaneous consideration of co-occurring parasites, host traits, and the environmental context will improve the understanding of host - parasite interactions and their consequences.

Linking sewage pollution and water quality to spatial patterns of Porites lobata growth anomalies in Puako, Hawaii.

Sewage pollution threatens the health of coastal populations and ecosystems, including coral reefs. We investigated spatial patterns of sewage pollution in Puako, Hawaii using enterococci concentrations and δ(15)N Ulva fasciata macroalgal bioassays to assess relationships with the coral disease Porites lobata growth anomalies (PGAs). PGA severity and enterococci concentrations were high, spatially variable, and positively related. Bioassay algal δ(15)N showed low sewage pollution at the reef edge while high values of resident algae indicated sewage pollution nearshore. Neither δ(15)N metric predicted PGA measures, though bioassay δ(15)N was negatively related to coral cover. Furthermore, PGA prevalence was much higher than previously recorded in Hawaii and the greater Indo-Pacific, highlighting Puako as an area of concern. Although further work is needed to resolve the relationship between sewage pollution and coral cover and disease, these results implicate sewage pollution as a contributor to diminished reef health.

Up in Arms: Immune and Nervous System Response to Sea Star Wasting Disease.

Echinoderms, positioned taxonomically at the base of deuterostomes, provide an important system for the study of the evolution of the immune system. However, there is little known about the cellular components and genes associated with echinoderm immunity. The 2013-2014 sea star wasting disease outbreak is an emergent, rapidly spreading disease, which has led to large population declines of asteroids in the North American Pacific. While evidence suggests that the signs of this disease, twisting arms and lesions, may be attributed to a viral infection, the host response to infection is still poorly understood. In order to examine transcriptional responses of the sea star Pycnopodia helianthoides to sea star wasting disease, we injected a viral sized fraction (0.2 µm) homogenate prepared from symptomatic P. helianthoides into apparently healthy stars. Nine days following injection, when all stars were displaying signs of the disease, specimens were sacrificed and coelomocytes were extracted for RNA-seq analyses. A number of immune genes, including those involved in Toll signaling pathways, complement cascade, melanization response, and arachidonic acid metabolism, were differentially expressed. Furthermore, genes involved in nervous system processes and tissue remodeling were also differentially expressed, pointing to transcriptional changes underlying the signs of sea star wasting disease. The genomic resources presented here not only increase understanding of host response to sea star wasting disease, but also provide greater insight into the mechanisms underlying immune function in echinoderms.

Persistent shifts in Caribbean coral microbiota are linked to the 2010 warm thermal anomaly.

The response of corals to warm temperature anomalies includes changes in coral bacterial assemblages. There are clear differences between the microbiota of bleached and healthy corals. However, few studies have tracked the microbiota of individual colonies throughout a warming event. We used 454 pyrosequencing and repeated measures to characterize bacterial assemblages in 15 Gorgonia ventalina colonies before, during, 4 months after, and 1 year after the 2010 Caribbean warm thermal anomaly. In the latter three sampling times, the G. ventalina microbiota differed significantly from the microbiota of Orbicella faveolata colonies, which were sampled only at these three times. O. faveolata microbiota did not exhibit coordinated shifts through time. Notably, the microbiota of the repeatedly sampled G. ventalina colonies shifted persistently from before to during, after, and long after the warming event. The same pattern emerges from the norm of reaction for the individual G. ventalina colonies. This is the first study to show persistent shifts in coral microbiota in association with a warm thermal anomaly. Whether shifting microbiota is adaptive or maladaptive, the lasting change in bacterial assemblages following this warming event identifies a new way that coral microbiota shape the response of coral colonies under thermal stress.

Comparing Sexuality Communication Among Offspring of Teen Parents and Adult Parents: a Different Role for Extended Family.

This brief report examined teenagers' sexuality communication with their parents and extended families. It compared who teens of early parents (those who had children when they were adolescents) and teens of later parents (those who were adults when they had children) talk to about sex. Eighth grade students (N=1281) in 24 schools completed survey items about their communication about sex. Structural equation modeling was used to predict communication profiles, while adjusting for the nesting of students within schools. After controlling for teens' age, gender, race/ethnicity, grades, parent/guardian closeness, and social desirability of survey responses, as well as family status and median family income, results showed that teens of early (teen) parents were more likely than teens of later (adult) parents to talk with both parents and extended family about sex and less likely than later parents to talk only with parents. These findings indicate that realities of teen sexuality communication for teens of early parents may extend beyond a parent-teen model to include extended family. Extended family involvement in educational outreach is a potential untapped resource to support sexual health for teens of early parents.

The role of extended family in diverse teens' sexual health.

Despite increasing extended family involvement in childrearing, particularly in minority families, few studies investigate their role in talking with teens about sex or how this relates to teens' sexual behavior. This mixed methods study assesses extended family sexuality communication through a survey of 1492 diverse middle school students and interviews with 32 students. Logistic regression shows that participants who report having had sex are more likely to report talking with extended family than those who report not having had sex. Interview themes explored reasons for and content of teen sexuality conversations with extended family. More sexually active teens' reporting communication with extended family is interpreted as extended family members gaining importance in sexuality communication as teens become sexually active.