Potential Use for Serosurveillance of Feral Swine to Map Risk for Anthrax Exposure, Texas, USA.

Anthrax is a disease of concern in many mammals, including humans. Management primarily consists of prevention through vaccination and tracking clinical-level observations because environmental isolation is laborious and bacterial distribution across large geographic areas difficult to confirm. Feral swine (Sus scrofa) are an invasive species with an extensive range in the southern United States that rarely succumbs to anthrax. We present evidence that feral swine might serve as biosentinels based on comparative seroprevalence in swine from historically defined anthrax-endemic and non-anthrax-endemic regions of Texas. Overall seropositivity was 43.7% (n = 478), and logistic regression revealed county endemicity status, age-class, sex, latitude, and longitude were informative for predicting antibody status. However, of these covariates, only latitude was statistically significant (ß = -0.153, p = 0.047). These results suggests anthrax exposure in swine, when paired with continuous location data, could serve as a proxy for bacterial presence in specific areas.

Population genomic transformations induced by isolation of wild bird avian influenza viruses (Orthomyxoviridae) in embryonated chicken eggs.

Isolation and cultivation of wild-type viruses in model organism cells or tissues is standard practice in virology. Oftentimes, the virus host species is distantly related to the species from which the culture system was developed. Thus, virus culture in these tissues and cells basically constitutes a host jump, which can lead to genomic changes through genetic drift and/or adaptation to the culture system. We directly sequenced 70 avian influenza virus (Orthomyxoviridae) genomes from oropharyngeal/cloacal swabs collected from wild bird species and paired virus isolates propagated from the same samples following isolation in specific-pathogen-free embryonated chicken eggs. The data were analyzed using population genetic approaches including evaluation of single nucleotide polymorphism (SNP) frequencies and divergence with pooled-sequencing analyses, consensus sequence placement in neighbor-joining trees, and haplotype reconstruction and networks. We found that propagation of virus in eggs leads to skewed SNP mutation spectra with some SNPs going to fixation. Both synonymous and nonsynonmous SNP frequencies shifted. We found multiple consensus sequences that differed between the swabs and the isolates, with some sequences from the same sample falling into divergent genetic clusters. Twenty of 23 coinfections detected had different dominant subtypes following virus isolation, thus sequences from both the swab and isolate were needed to obtain full subtype data. Haplotype networks revealed haplotype frequency shifts and the appearance or loss of low-frequency haplotypes following isolation. The results from this study revealed that isolation of wild bird avian influenza viruses in chicken eggs leads to skewed populations that are different than the input populations. Consensus sequence changes from virus isolation can lead to flawed phylogenetic inferences, and subtype detection is biased. These results suggest that for genomic studies of wild bird influenza viruses the biological field should move away from chicken egg isolation towards directly sequencing the virus from host samples.

Overexpression of AtAHL20 causes delayed flowering in Arabidopsis via repression of FT expression.

BACKGROUND: The 29-member Arabidopsis AHL gene family is classified into three main classes based on nucleotide and protein sequence evolutionary differences. These differences include the presence or absence of introns, type and/or number of conserved AT-hook and PPC domains. AHL gene family members are divided into two phylogenetic clades, Clade-A and Clade-B. A majority of the 29 members remain functionally uncharacterized. Furthermore, the biological significance of the DNA and peptide sequence diversity, observed in the conserved motifs and domains found in the different AHL types, is a subject area that remains largely unexplored. RESULTS: Transgenic plants overexpressing AtAHL20 flowered later than the wild type under both short and long days. Transcript accumulation analyses showed that 35S:AtAHL20 plants contained reduced FT, TSF, AGL8 and SPL3 mRNA levels. Similarly, overexpression of AtAHL20's orthologue in Camelina sativa, Arabidopsis' closely related Brassicaceae family member species, conferred a late-flowering phenotype via suppression of CsFT expression. However, overexpression of an aberrant AtAHL20 gene harboring a missense mutation in the AT-hook domain's highly conserved R-G-R core motif abolished the late-flowering phenotype. Data from targeted yeast-two-hybrid assays showed that AtAHL20 interacted with itself and several other Clade-A Type-I AHLs which have been previously implicated in flowering-time regulation: AtAHL19, AtAHL22 and AtAHL29. CONCLUSION: We showed via gain-of-function analysis that AtAHL20 is a negative regulator of FT expression, as well as other downstream flowering time regulating genes. A similar outcome in Camelina sativa transgenic plants overexpressing CsAHL20 suggest that this is a conserved function. Our results demonstrate that AtAHL20 acts as a photoperiod-independent negative regulator of transition to flowering.

Loci Associated With Antibody Response in Feral Swine (Sus scrofa) Infected With Brucella suis.

Feral swine (Sus scrofa) are a destructive invasive species widespread throughout the United States that disrupt ecosystems, damage crops, and carry pathogens of concern for the health of domestic stock and humans including Brucella suis-the causative organism for swine brucellosis. In domestic swine, brucellosis results in reproductive failure due to abortions and infertility. Contact with infected feral swine poses spillover risks to domestic pigs as well as humans, companion animals, wildlife, and other livestock. Genetic factors influence the outcome of infectious diseases; therefore, genome wide association studies (GWAS) of differential immune responses among feral swine can provide an understanding of disease dynamics and inform management to prevent the spillover of brucellosis from feral swine to domestic pigs. We sought to identify loci associated with differential antibody responses among feral swine naturally infected with B. suis using a case-control GWAS. Tissue, serum, and genotype data (68,516 bi-allelic single nucleotide polymorphisms) collected from 47 feral swine were analyzed in this study. The 47 feral swine were culture positive for Brucella spp. Of these 47, 16 were antibody positive (cases) whereas 31 were antibody negative (controls). Single-locus GWAS were performed using efficient mixed-model association eXpedited (EMMAX) methodology with three genetic models: additive, dominant, and recessive. Eight loci associated with seroconversion were identified on chromosome 4, 8, 9, 10, 12, and 18. Subsequent bioinformatic analyses revealed nine putative candidate genes related to immune function, most notably phagocytosis and induction of an inflammatory response. Identified loci and putative candidate genes may play an important role in host immune responses to B. suis infection, characterized by a detectable bacterial presence yet a differential antibody response. Given that antibody tests are used to evaluate brucellosis infection in domestic pigs and for disease surveillance in invasive feral swine, additional studies are needed to fully understand the genetic component of the response to B. suis infection and to more effectively translate estimates of Brucella spp. antibody prevalence among feral swine to disease control management action.

Synopsis of the SOFL Plant-Specific Gene Family.

SUPPRESSOR OF PHYB-4#5DOMINANT (sob5-D) was previously identified as a suppressor of the phyB-4 long-hypocotyl phenotype in Arabidopsis thaliana Overexpression of SOB5 conferred dwarf phenotypes similar to those observed in plants containing elevated levels of cytokinin (CK) nucleotides and nucleosides. Two SOB-FIVE- LIKE (SOFL) proteins, AtSOFL1 and AtSOFL2, which are more similar at the protein level to each other than they are to SOB5, conferred similar phenotypes to the sob5-D mutant when overexpressed. We used protein sequences of founding SOFL gene family members to perform database searches and identified a total of 289 SOFL homologs in genomes of 89 angiosperm species. Phylogenetic analysis results implied that the SOFL gene family emerged during the expansion of angiosperms and later evolved into four distinct clades. Among the newly identified gene family members are four previously unreported Arabidopsis SOFLs Multiple sequence alignment of the 289 SOFL protein sequences revealed two highly conserved domains; SOFL-A and SOFL-B. We used overexpression and site-directed mutagenesis studies to demonstrate that SOFL domains are necessary for SOB5 and AtSOFL1's overexpression phenotypes. Examination of the subcellular localization patterns of founding Arabidopsis thaliana SOFLs suggested they may be localized in the cytoplasm and/or the nucleus. Overall, we report that SOFLs are a plant-specific gene family characterized by two conserved domains that are important for function.

Opioid overdose prevention with intranasal naloxone among people who take methadone.

Overdose education and naloxone distribution (OEND) is an intervention that addresses overdose, but has not been studied among people who take methadone, a drug involved in increasing numbers of overdoses. This study describes the implementation of OEND among people taking methadone in the previous 30 days in various settings in Massachusetts. From 2008 to 2010, 1553 participants received OEND who had taken methadone in the past 30 days. Settings included inpatient detoxification (47%), HIV prevention programs (25%), methadone maintenance treatment programs (MMTP) (17%), and other settings (11%). Previous overdose, recent inpatient detoxification and incarceration, and polysubstance use were overdose risks factors common among all groups. Participants reported 92 overdose rescues. OEND programs are public health interventions that address overdose risk among people who take methadone and their social networks. OEND programs can be implemented in MMTPs, detoxification programs, and HIV prevention programs.

Methadone dose, take home status, and hospital admission among methadone maintenance patients.

OBJECTIVES: Among patients receiving methadone maintenance treatment (MMT) for opioid dependence, receipt of unobserved dosing privileges (take homes) and adequate doses (ie, ≥ 80 mg) are each associated with improved addiction treatment outcomes, but the association with acute care hospitalization is unknown. We studied whether take-home dosing and adequate doses (ie, ≥80 mg) were associated with decreased hospital admission among patients in an MMT. METHODS: We reviewed daily electronic medical records of patients enrolled in one MMT program to determine receipt of take-home doses, methadone dose 80 mg or more, and hospital admission date. Nonlinear mixed-effects logistic regression models were used to evaluate whether take-home doses or dose 80 mg or more on a given day were associated with hospital admission on the subsequent day. Covariates in adjusted models included age, sex, race/ethnicity, human immunodeficiency virus status, medical illness, mental illness, and polysubstance use at program admission. RESULTS: Subjects (n = 138) had the following characteristics: mean age 43 years; 52% female; 17% human immunodeficiency virus-infected; 32% medical illness; 40% mental illness; and 52% polysubstance use. During a mean follow-up of 20 months, 42 patients (30%) accounted for 80 hospitalizations. Receipt of take homes was associated with significantly lower odds of a hospital admission (adjusted odds ratio [AOR] = 0.26; 95% confidence interval [CI], 0.11-0.62), whereas methadone dose 80 mg or more was not (AOR = 1.01; 95% CI, 0.56-1.83). CONCLUSIONS: Among MMT patients, receipt of take homes, but not dose of methadone, was associated with decreased hospital admission. Take-home status may reflect not only patients' improved addiction outcomes but also reduced health care utilization.

Evaluation of a digitally integrated accelerometer-based activity monitor for the measurement of activity in cats.

OBJECTIVE: To assess the correlation between data generated by an accelerometer-based activity monitor and the distance moved in cats. STUDY DESIGN: Prospective experimental study. ANIMALS: Three, four-year-old, male, purpose-bred research cats, weighing between 5.1 and 5.9 kg. METHODS: Part I: Collar and harness mounted accelerometers were evaluated in three cats, comparing simultaneously collected accelerometer data with movement data from computer-analyzed video. Part II: Cats wore collar and harness mounted accelerometers, and data were recorded for 4 weeks to evaluate day-to-day and week-to-week variation in activity. RESULTS: Part I: 432 hours of simultaneous video and accelerometer data were collected. The correlation between accelerometer counts and distance moved was 0.82 overall. Agreement between collar and harness mounted accelerometers was excellent with only 6% of the differences in measurements lying outside the mean difference +/- 2 standard deviations. The adjusted R(2) for harness accelerometer output and 6% mobility was 0.75; for movement 0.84; and for mean velocity 0.83. Evaluation of video indicated eating, grooming and scratching created high accelerometer counts with little effect on movement. Part II: There was a significant effect of day on harness (p < 0.001) and collar (p < 0.002) counts, with counts being lowest at the weekend. There was a significant effect of week on harness-mounted accelerometer counts (p < 0.034), but not on collar-mounted accelerometer counts. Harness accelerometer counts were lowest in week 1. CONCLUSION: Output from an acceleration-based digitally integrated accelerometer correlated well with distance moved and mobility in freely moving cats provided the mobility threshold in the analysis software was > or = 6%. CLINICAL RELEVANCE: Acceleration-based activity monitors may allow for objective measurement of improved mobility following analgesic treatment for conditions such as osteoarthritis.

Evaluation of client-specific outcome measures and activity monitoring to measure pain relief in cats with osteoarthritis.

BACKGROUND: There are no validated systems for measuring pain from osteoarthritis in cats. HYPOTHESIS: Owner subjective assessments and an activity monitor (AM) can be used to detect pain in cats with osteoarthritis and to assess efficacy of treatments. ANIMALS: Thirteen cats older than 10 years old, with owner-assessed decreases in activity, painful arthritic joints, and clinically normal blood work were included and evaluated for 3 weeks. METHODS: A collar-mounted AM measured activity and a client-specific outcome measure (CSOM) questionnaire characterized the severity of impairment. Overall global quality of life was also evaluated for each treatment. In weeks 2 and 3, meloxicam (0.1 mg/kg, day 1; 0.05 mg/kg, days 2-5) or a placebo was administered in a blinded, randomized, cross-over manner to test the assessment systems. RESULTS: The cats had a median of 4 arthritic appendicular joints. Activity counts for the week when cats (complete data on activity; n=9) were administered meloxicam were significantly higher than at baseline (P = .02) but not after placebo (P = .06). Baseline activity counts were not significantly different from placebo (P = .6). The CSOM data (n=13) showed that owners considered their cats to be more active on meloxicam compared with baseline (P = .001) and placebo (P < .004), and more active on placebo than at baseline (P < .01). Global quality of life improved significantly with meloxicam (P < .042). CONCLUSIONS AND CLINICAL IMPORTANCE: Both an AM and a CSOM system can detect behavior associated with pain relief in cats that are arthritic. Objective activity data might allow subjective assessment systems to be validated for use in clinical studies.

Examining subtypes of behavior problems among 3-year-old children, Part II: investigating differences in parent psychopathology, couple conflict, and other family stressors.

This study examined family stressors among 3-year-old children who were classified as hyperactive (HYP), hyperactive and oppositional defiant (HYP/OD), and non-problem based on mothers' reports of behavior. Children with HYP/OD were found to experience higher levels of family stressors than non-problem children on almost every family stressor variable. Compared to children with HYP, families of children with HYP/OD also tended to report more Axis II maternal psychopathology, Axis I paternal psychopathology, and high intensity couple conflict tactics. However, the HYP and HYP/OD group did not significantly differ on maternal Axis I psychopathology, paternal Axis II psychopathology, parental marital status, negative life events, frequency of couple conflict, or use of lower intensity couple conflict tactics. Parents of children with HYP and HYP/OD reported more negative life events, more maternal adult ADHD symptoms, and more maternal avoidance and verbal aggression during marital conflict than parents of non-problem children. Implications for treatment and etiology are discussed.

Shift Work, Role Overload, and the Transition to Parenthood.

This article examines how the work hours, work schedules, and role overload of working-class couples are related to depressive symptoms and relationship conflict across the transition to parenthood. Data are from 132 dual-earner couples interviewed 5 times across the transition. Multilevel modeling analyses revealed that working evening or night shifts, as opposed to day shifts, was related to higher levels of depressive symptoms. For mothers only, working rotating shifts predicted relationship conflict. Increases in role overload were positively related to both depression and conflict; working a nonday shift explained variance in depression and conflict above and beyond role overload. Results suggest that for new parents, working nonday shifts may be a risk factor for depressive symptoms and relationship conflict.