Development of duplex qPCR targeting Carnobacterium maltaromaticum and Vagococcus salmoninarum.

In November 2018, Vagococcus salmoninarum was identified as the causative agent of a chronic coldwater streptococcosis epizootic in broodstock brook trout (Salvelinus fontinalis) at the Iron River National Fish Hatchery in Wisconsin, USA. By February 2019, the epizootic spread to adjacent raceways containing broodstock lake trout (Salvelinus namaycush), whereby fish were found to be coinfected with Carnobacterium maltaromaticum and V. salmoninarum. To differentiate these two pathogens and determine the primary cause of the lake trout morbidity, a quantitative real-time PCR (qPCR) was developed targeting the C. maltaromaticum phenylalanyl-tRNA synthase alpha subunit (pheS) gene. The qPCR was combined with a V. salmoninarum qPCR, creating a duplex qPCR assay that simultaneously quantitates C. maltaromaticum and V. salmoninarum concentrations in individual lake trout tissues, and screens presumptive isolates from hatchery inspections and wild fish from national fish hatchery source waters throughout the Great Lakes basin. Vagococcus salmoninarum and C. maltaromaticum were co-detected in broodstock brook trout from two tribal hatcheries and C. maltaromaticum was present in wild fish in source waters of several national fish hatcheries. This study provides a powerful new tool to differentiate and diagnose two emerging Gram-positive bacterial pathogens.

Isolations of the Spring Viremia of Carp Virus in the Upper Mississippi River (USA), Including a New Host, the Quillback.

In July of 2018 and 2019, wild fish health surveys were conducted along the Wisconsin and Minnesota portions of the upper Mississippi River. Spring viremia of carp virus (SVCV) was isolated from Common Carp Cyprinus carpio as well as a newly identified host species, the Quillback Carpiodes cyprinus. Sanger sequencing of the gene encoding for the G protein revealed a high similarity of the Quillback isolate to various SVCV isolates identified from Common Carp that were collected during earlier wild fish health surveys and mortality events in the USA. Despite annual monitoring, this virus has been infrequently identified. The speculative role of native fish and invertebrates in allowing the virus to persist for long periods without detection is discussed.

Childhood Loss Experiences, Religiousness/Spirituality, and Hope as Predictors of Adult Life Satisfaction.

Purpose The purpose of this study was to explore how adaptive patterns of religiousness/spirituality and hope predict adult life satisfaction in adults, even if they had childhood loss experiences.Design and Methods Using a cross-sectional survey design with132 adult participants (mean age 33.8 ± 15.2, 82% female, 78% Caucasian) we estimated two hierarchical regression models to examine how dimensions of religiousness/spirituality (Brief Multidimensional Measure of Religiousness/Spirituality) and hope (Herth Hope Index) predicted adult life satisfaction (Satisfaction with Life Scale) after controlling for effects of adverse childhood experiences (Adverse Childhood Experiences Study Questionnaire) or chronic sorrow from childhood loss (Kendall Chronic Sorrow Instrument).Results When effects of adverse childhood experiences (ACE) were controlled, higher levels of hope (ß = .416, p < .001) and forgiveness (ß = .273, p = .023) and lower levels of religious/spiritual values/beliefs (ß = -.298, p = .014) predicted higher levels of adult life satisfaction. ACE remained a significant predictor of adult life satisfaction in the final model. When controlling for effects of chronic sorrow, higher life satisfaction was predicted only by forgiveness (ß = .379, p = .003), values/beliefs (ß = -.354, p = .007), and hope (ß = .357, p < .001), with chronic sorrow not a significant predictor of adult life satisfaction in the final model.Conclusion: Greater life satisfaction for adult survivors of childhood loss experiences could be promoted by interventions to enhance hope and foster forgiveness. These adult survivors may also need support as they engage with religious/spiritual struggle to reconcile their values and beliefs with childhood loss.

Vagococcus salmoninarum II-qPCR, tropism and egg-associated transmission.

Vagococcus salmoninarum was identified as the causative agent of a chronic epizootic in broodstock "coaster" brook trout (Salvelinus fontinalis) at the Iron River National Fish Hatchery. The epizootic spanned more than a year, was unresponsive to multiple florfenicol treatments, and resulted in >50% mortality of the affected fish. The decision was made to cull the remaining fish during spawning, which presented an opportunity to more thoroughly examine V. salmoninarum sampling methods, organ tropism and vertical transmission. A newly developed qPCR targeting the pheS gene was used in concert with bacterial culture to show that V. salmoninarum indeed disproportionately affects females and has a tropism for female reproductive tissues. The study demonstrates that some female reproductive tissues (e.g. ovarian fluid, unfertilized eggs) are also an effective option for non-lethal detection. Despite the widespread presence of V. salmoninarum in ovarian fluid and on egg surfaces, we found no evidence of intra-ova transmission.

Vagococcus salmoninarum I-A chronic coldwater streptococcosis in broodstock brook trout (Salvelinus fontinalis) in Wisconsin, USA.

In 2018, Vagococcus salmoninarum was isolated from two lots of broodstock "coaster" brook trout (Salvelinus fontinalis) containing ~1,500 fish at the Iron River National Fish Hatchery, at which time it was identified as the causative agent of a chronic coldwater streptococcosis epizootic. Clinical signs included exophthalmia, lethargy, erratic swimming and loss of equilibrium. Female fish experienced disproportionately higher morbidity and mortality than male co-inhabitants, and routinely retained eggs following spawning. The most consistent gross clinical sign was heart pallor and turbid pericardial effusion. An attempted treatment using florfenicol was ineffective at halting the epizootic, which spanned more than a year and resulted in >50% mortality before remaining fish were culled. As there is no previous documentation of V. salmoninarum at this hatchery or in this species, it is still unclear what circumstances led to this epizootic. The inability to treat this chronic disease led to the loss of valuable broodstock, hampering ongoing fishery conservation efforts in the Great Lakes Basin.

Optimizing, validating, and field testing a multiplex qPCR for the detection of amphibian pathogens.

Amphibian populations worldwide are facing numerous threats, including the emergence and spread of infectious diseases. In the past 2 decades, Batrachochytrium dendrobatidis (Bd), a parasitic fungus, and a group of viruses comprising the genus Ranavirus have become widespread and resulted in mass mortality events and extirpations worldwide. In 2013, another novel fungus, B. salamandrivorans (Bsal), was attributed to dramatic declines in populations of fire salamander Salamandra salamandra in the Netherlands. Experimental infections demonstrated that Bsal is highly pathogenic to numerous salamander genera. In an effort to prevent the introduction of Bsal to North America, the US Fish and Wildlife Service (USFWS) listed 201 salamander species as injurious wildlife under the Lacey Act. To determine infection status and accurately assess amphibian health, the development of a sensitive and specific diagnostic assay was needed. We describe the optimization and validation of a multiplex quantitative polymerase chain reaction (qPCR) protocol for the simultaneous detection of Bd, Bsal, and frog virus 3-like ranaviruses. A synthetic genome template (gBlock®) containing the target genes from all 3 pathogens served as the positive control and allowed accurate quantification of pathogen genes. The assay was validated in the field using an established non-lethal swabbing technique to survey local amphibian populations throughout a range of habitats. This multiplex qPCR demonstrates high reproducibility, sensitivity, and was capable of detecting both Bd and ranavirus in numerous locations, species, and life stages. Bsal was not detected at any point during these sampling efforts.

Psychotherapy Experiences of Perpetrators of Child Sexual Abuse.

Cynicism about treatment of sex offenders pervades both professional and lay literature. A Cochrane review of randomized controlled trials concluded there is no evidence to support any psychological intervention for sex offenders, but RCT design has limitations for evaluating sex offender treatment. Rarely has a qualitative approach been used to explore perceptions of offenders themselves about their psychotherapy experiences. The purpose of this study was to discover the meaning of therapy experiences to 11 community-dwelling perpetrators of child sexual abuse. They had received therapy during incarceration or after release, or both. Secondary analysis was conducted of phenomenological interviews about participants' early life, during which they spontaneously revealed insights gleaned during therapy in adulthood. Rigor of the analysis was enhanced by reading transcripts aloud and thematizing them in an interdisciplinary interpretive group. Five interrelated themes constituted a gestalt comprising the essence of the therapy experience: "This treatment, it's just totally changed my whole world." Themes included: "It just stripped away all the pretense, all the lies, all the manipulation;" "I didn't understand myself; I found out all about myself through this;" "Nobody knew any of my secrets; that (therapy) was the first time that I got to tell my story;" "The group has become a family for me;" and "I'm very ashamed of what I've done; this treatment has really helped me, gave me a second chance." These findings stand in contrast to cynicism about sex offender therapy and lend support to the increased optimism expressed by several contemporary scholars.

Childhood sibling and peer relationships of perpetrators of child sexual abuse.

Maltreatment by parents inhibits a child's ability to form relationships with other children. Given that many sex offenders experienced parental abuse, investigation of their relationships with siblings and peers could provide new directions for early childhood interventions. Interview transcripts of 23 community-dwelling perpetrators of sexual abuse against children were examined in this secondary data analysis. Ages of participants ranged from 20s to 70s, and education ranged from GED to PhD. Phenomenological analysis revealed that neither sibling nor peer relationships adequately promoted participants' capacity for developing and maintaining healthy relationships in adulthood. Until societal preventive actions and early clinical interventions are more successful, the crime of child sexual abuse will continue to be far too common.

Hearing the silent voices: narratives of health care and homelessness.

Most homeless individuals lack adequate health care. With existing literature as a backdrop, this study sought to understand the experience of homeless persons in the health care system. Using a phenomenological approach, 11 homeless participants were interviewed and the transcripts from these interviews were analyzed for meaning. The health care experiences of the participants could be understood only when viewed within the context of homelessness. The four polar themes that emerged from the analysis--same/different, fair/unfair, freedom/barriers, and choice/no choice--highlighted the great divide between the health care experiences of those with a home and those without. Such understanding can help mental health nurses provide more appropriate care to this population.

Dissolvable alginate hydrogel-based biofilm microreactors for antibiotic susceptibility assays.

Biofilms are found in many infections in the forms of surface-adhering aggregates on medical devices, small clumps in tissues, or even in synovial fluid. Although antibiotic resistance genes are studied and monitored in the clinic, the structural and phenotypic changes that take place in biofilms can also lead to significant changes in how bacteria respond to antibiotics. Therefore, it is important to better understand the relationship between biofilm phenotypes and resistance and develop approaches that are compatible with clinical testing. Current methods for studying antimicrobial susceptibility are mostly planktonic or planar biofilm reactors. In this work, we develop a new type of biofilm reactor-three-dimensional (3D) microreactors-to recreate biofilms in a microenvironment that better mimics those in vivo where bacteria tend to form surface-independent biofilms in living tissues. The microreactors are formed on microplates, treated with antibiotics of 1000 times of the corresponding minimal inhibitory concentrations (1000 × MIC), and monitored spectroscopically with a microplate reader in a high-throughput manner. The hydrogels are dissolvable on demand without the need for manual scraping, thus enabling measurements of phenotypic changes. Bacteria inside the biofilm microreactors are found to survive exposure to 1000 × MIC of antibiotics, and subsequent comparison with plating results reveals no antibiotic resistance-associated phenotypes. The presented microreactor offers an attractive platform to study the tolerance and antibiotic resistance of surface-independent biofilms such as those found in tissues.

Emergence of Viral hemorrhagic septicemia virus in the North American Great Lakes region is associated with low viral genetic diversity.

Viral hemorrhagic septicemia virus (VHSV) is a fish rhabdovirus that causes disease in a broad range of marine and freshwater hosts. The known geographic range includes the Northern Atlantic and Pacific Oceans, and recently it has invaded the Great Lakes region of North America. The goal of this work was to characterize genetic diversity of Great Lakes VHSV isolates at the early stage of this viral emergence by comparing a partial glycoprotein (G) gene sequence (669 nt) of 108 isolates collected from 2003 to 2009 from 31 species and at 37 sites. Phylogenetic analysis showed that all isolates fell into sub-lineage IVb within the major VHSV genetic group IV. Among these 108 isolates, genetic diversity was low, with a maximum of 1.05% within the 669 nt region. There were 11 unique sequences, designated vcG001 to vcG011. Two dominant sequence types, vcG001 and vcG002, accounted for 90% (97 of 108) of the isolates. The vcG001 isolates were most widespread. We saw no apparent association of sequence type with host or year of isolation, but we did note a spatial pattern, in which vcG002 isolates were more prevalent in the easternmost sub-regions, including inland New York state and the St. Lawrence Seaway. Different sequence types were found among isolates from single disease outbreaks, and mixtures of types were evident within 2 isolates from individual fish. Overall, the genetic diversity of VHSV in the Great Lakes region was found to be extremely low, consistent with an introduction of a new virus into a geographic region with previously naive host populations.

Development of an instrument to measure internalized stigma in those with HIV/AIDS.

Stigma has grave consequences for persons living with HIV/AIDS. Stigma hampers prevention of HIV transmission to sexual partners and to unborn babies, diagnosis, and early treatment, and negatively affects mental and physical health, quality of life, and life satisfaction. Internalized stigma of HIV/AIDS may have even more severe consequences than perceived or enacted stigma. The purpose of this study was to develop an instrument to measure internalized stigma in those with HIV/AIDS. Data were drawn from the Rural Women's Health Project. Research assistants administered structured interviews at baseline, 3 months, and 6 months. Instruments used in these analyses included a demographic data form, the Centers for Epidemiological Studies Depression Scale (CES-D), the Perceived Stigma Scale (PSS), and the Internalized Stigma of AIDS Tool (ISAT). Exploratory factor analysis confirmed that the ten items of the ISAT measure a single factor that explains 88% of the variance in the construct. Internal consistency was demonstrated by a Cronbach's alpha of .91 (Time 1), .92 (Time 2), and .92 (Time 3). Convergent validity was supported with significant positive correlations with the CES-D (rho = 0.33, p < 0.0001) and the PSS (rho = 0.56, < 0.0001). The Internalized Stigma of AIDS Tool appears to be a reliable and valid instrument to measure internalization of the stigma of HIV/AIDS. It may be of value in research and clinical assessment.

Social context of rural Women with HIV/AIDS.

The South has more AIDS cases than any other region of the US, with most new diagnoses among African American women (56%). In a previous study, a peer counseling intervention for rural women with HIV/AIDS was developed and tested. The purpose of this analysis was to describe, from the peer counselors' perspective, the predominant concerns of the women, contextualized by living in isolated, impoverished circumstances in the rural Deep South. Following home visits, peer counselors recorded a description of the encounter. A multidisciplinary qualitative research group extracted, coded, and thematized the chief concerns and context of the women's lives. Findings provide a vivid portrait of HIV-infected women experiencing deeply troubling psychological and physiological symptoms of HIV/AIDS against the contextual ground of poverty and isolation. Themes include: (1) struggle/effort; (2) stigma/hiding; (3) loss/depression; and (4) independence/ dependence. These women lived in extremely difficult life circumstances that reflected not only a devastating chronic illness, but a life of poverty and abuse. Appropriate care for HIV-infected women living in the rural Deep South will need to address the whole context of their lives.

Dynamic Adaptive Response of Pseudomonas aeruginosa to Clindamycin/Rifampicin-Impregnated Catheters.

Pseudomonas aeruginosa is the most common Gram-negative pathogen causing nosocomial multidrug resistant infections. It is a good biofilm producer and has the potential for contaminating medical devices. Despite the widespread use of antibacterial-impregnated catheters, little is known about the impacts of antibacterial coating on the pathogenesis of P. aeruginosa. In this study, we investigated the adaptive resistance potential of P. aeruginosa strain PAO1 in response to continuous antibiotic exposure from clindamycin/rifampicin-impregnated catheters (CR-IC). During exposure for 144 h to clindamycin and rifampicin released from CR-IC, strain PAO1 formed biofilms featuring elongated and swollen cells. There were 545 and 372 differentially expressed proteins (DEPs) identified in the planktonic and biofilm cells, respectively, by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Both Cluster of Orthologous Groups (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that the planktonic cells responded to the released antibiotics more actively than the biofilm cells, with metabolism and ribosomal biosynthesis-associated proteins being significantly over-expressed. Exposure to CR-IC increased the invasion capability of P. aeruginosa for Hela cells and upregulated the expression of certain groups of virulence proteins in both planktonic and biofilm cells, including the outer membrane associated (flagella, type IV pili and type III secretion system) and extracellular (pyoverdine) virulence proteins. Continuous exposure of P. aeruginosa to CR-IC also induced the overexpression of antibiotic resistance proteins, including porins, efflux pumps, translation and transcription proteins. However, these upregulations did not change phenotypic minimum inhibitory concentration (MIC) during the experimental timeframe. The concerning association between CR-IC and overexpression of virulence factors in P. aeruginosa suggests the need for additional investigation to determine if it results in adverse clinical outcomes.

Men seeking sex on an intergenerational gay Internet website: an exploratory study.

OBJECTIVE: This study explored the characteristics and sexual behaviors reported by men who have sex with men (MSM) seeking sex on an intergenerational website. Of special interest was to determine whether and how seeking sex on the Internet contributed to risky sex behaviors. METHODS: This descriptive exploratory study extracted data from a stratified random sample of 1,020 profiles posted by men seeking sex on a gay intergenerational website. Frequencies and percentages were calculated for all categorical variables. Analysis of variance was used to test for differences in subgroup categories. RESULTS: The men in the sample were primarily Caucasian (92.3%) with a mean age of 50 years (range: 18-88 years). More than one-fourth of the men (28.5%) reported being married. Of the married men, 76.0% indicated they would engage in receptive anal intercourse. Only a small number of men in the total sample expressed interest in safe sex (17.5%), while 91 men (8.9%) directly stated that they wanted unsafe sex. Most men in the sample were seeking intergenerational sexual encounters, with 71.0% of younger men expressing a preference for older men. CONCLUSIONS: The Internet can play a significant role in seeking same-sex encounters across generational groups. Safe sex was not a stated priority for most men in this study, and the Internet provides an effective method of seeking unsafe sex. A number of MSM seeking sex on the Internet were married and Caucasian. This finding suggests the need for greater attention to married and/or publicly identified Caucasian, heterosexual men in human immunodeficiency virus prevention efforts. Effective Internet-based prevention programs need to be implemented and researched.

Understanding and responding to HIV/AIDS stigma and disclosure: an international challenge for mental health nurses.

Stigma and discrimination are challenges in the care and treatment of persons with HIV infection worldwide. Fear of negative social consequences often causes persons with HIV/AIDS to keep their infection secret, resulting in negative psychological and physical outcomes and continued spread of the disease. Mental health nurses have a unique opportunity to influence the trajectory of HIV/AIDS though counseling and interventions that address HIV/AIDS stigma with clients, communities, and society. This article provides an in-depth examination of HIV/AIDS stigma and its relationship to nondisclosure as well as strategies to deal with these issues at individual and group levels.

Cranial electrotherapy stimulation for the treatment of depression.

More prevalent in women than men, clinical depression affects approximately 15 million American adults in a given year. Psychopharmaceutical therapy accompanied by psychotherapy and wellness interventions (e.g., nutrition, exercise, counseling) is effective in 80% of diagnosed cases. A lesser known adjunctive therapy is that of cranial electrotherapy stimulation (CES). The major hypothesis for the use of CES in depression is that it may reset the brain to pre-stress homeostasis levels. It is conjectured that the pulsed electrical currents emitted by cranial electrical stimulators affect changes in the limbic system, the reticular activating system, and/or the hypothalamus that result in neurotransmitter secretion and downstream hormone production. While evidence is good for applied research, basic research about the mechanisms of action for CES remains in its infancy. A review of the literature provides an overview of current research findings and implications for clinical mental health practice.

Rat toxicogenomic study reveals analytical consistency across microarray platforms.

To validate and extend the findings of the MicroArray Quality Control (MAQC) project, a biologically relevant toxicogenomics data set was generated using 36 RNA samples from rats treated with three chemicals (aristolochic acid, riddelliine and comfrey) and each sample was hybridized to four microarray platforms. The MAQC project assessed concordance in intersite and cross-platform comparisons and the impact of gene selection methods on the reproducibility of profiling data in terms of differentially expressed genes using distinct reference RNA samples. The real-world toxicogenomic data set reported here showed high concordance in intersite and cross-platform comparisons. Further, gene lists generated by fold-change ranking were more reproducible than those obtained by t-test P value or Significance Analysis of Microarrays. Finally, gene lists generated by fold-change ranking with a nonstringent P-value cutoff showed increased consistency in Gene Ontology terms and pathways, and hence the biological impact of chemical exposure could be reliably deduced from all platforms analyzed.

Embodied work: insider perspectives on the work of HIV/AIDS peer counselors.

Our aim in this study was to explore HIV/AIDS peer counseling from the perspective of women actively engaged in this work within the context of a community-based program in rural areas of the southeastern United States. Based on this research we suggest that the embodied work of HIV/AIDS peer counselors is constructed around their personal identities and experiences. This work involves gaining entry to other HIV-positive women's lives, building relationships, drawing on personal experiences, facing issues of fear and stigma, tailoring peer counseling for diversity, balancing risks and benefits, and terminating relationships. Peer counselors recognize the personal and collective value of their work, which, like much of women's work within the context of family and community, lacks public visibility and acknowledgment. We discuss implications for the training and support of peer-based interventions for HIV and other women's health issues across diverse contexts and settings.

Moderate intensity exercise training reverses functional aerobic impairment in HIV-infected individuals.

HIV infection and HIV drug therapies result in physical and psychological challenges to those living with HIV. These conditions contribute to decreased functional aerobic capacity (FAC). The aim of this study was to determine the effects of a combined moderate-intensity aerobic and resistance exercise intervention on the FAC of HIV-infected individuals. Forty HIV-infected individuals were randomized to an exercise group (EX) who completed six weeks of moderate-intensity exercise training, or to a control group (CON) that did not receive the exercise intervention. Twice weekly, the EX group completed 30 minutes of moderate-intensity aerobic training followed by moderate-intensity resistance training. Prior to, and following, the intervention the FAC for each subject was determined by graded exercise treadmill stress test (GXT). At baseline testing, the mean FAC as determined by treadmill time-based estimation of maximal oxygen consumption was 25% below age-predicted values, a level of reduction indicating the presence of functional aerobic impairment (FAI). Following the intervention, the EX had a significant increase in time to fatigue and estimated VO(2) max (p<.001). Further, FAI was eliminated (1% above age predicted values) during the exercise training. The EX group also experienced decreased heart rates during Stages 1 (p=.02), 2 (p=.01), 4 (p=.05) and 6 (p=.02) of the GXT. The CON had no significant changes during the intervention period. These data indicate that six weeks of combined moderate-intensity aerobic and resistance training can improve FAC and eliminate FAI in those with HIV. Results suggest that the functional limitations common in HIV-infected individuals are due in part to detraining that is reversible through moderate exercise adherence.