Multiplex evaluation of influenza neutralizing antibodies with potential applicability to in-field serological studies

The increased number of outbreaks of H5 and H7 LPAI and HPAI viruses in poultry has major public and animal health implications. The continuous rapid evolution of these subtypes and the emergence of new variants influence the ability to undertake effective surveillance. Retroviral pseudotypes bearing influenza haemagglutinin (HA) and neuraminidase (NA) envelope glycoproteins represent a flexible platform for sensitive, readily standardized influenza serological assays. We describe a multiplex assay for the study of neutralizing antibodies that are directed against both influenza H5 and H7 HA. This assay permits the measurement of neutralizing antibody responses against two antigenically distinct HAs in the same serum/plasma sample thus increasing the amount and quality of serological data that can be acquired from valuable sera. Sera obtained from chickens vaccinated with a monovalent H5N2 vaccine, chickens vaccinated with a bivalent H7N1/H5N9 vaccine, or turkeys naturally infected with an H7N3 virus were evaluated in this assay and the results correlated strongly with data obtained by HI assay. We show that pseudotypes are highly stable under basic cold-chain storage conditions and following multiple rounds of freeze-thaw. We propose that this robust assay may have practical utility for in-field sero-surveillance and vaccine studies in resource-limited regions worldwide

Vaccine Interaction and Protection against Virulent Avian Metapneumovirus (aMPV) Challenge after Combined Administration of Newcastle Disease and aMPV Live Vaccines to Day-Old Turkeys

Newcastle disease virus (NDV) and avian metapneumovirus (aMPV) are among the most impactful pathogens affecting the turkey industry. Since turkeys are routinely immunized against both diseases, the hatchery administration of the combined respective live vaccines would offer remarkable practical advantages. However, the compatibility of NDV and aMPV vaccines has not yet been experimentally demonstrated in this species. To address this issue, an aMPV subtype B live vaccine was administered to day-old poults either alone or in combination with one of two different ND vaccines. The birds were then challenged with a virulent aMPV subtype B strain, clinical signs were recorded and aMPV and NDV vaccine replication and humoral immune response were assessed. All results supported the absence of any interference hampering protection against aMPV, with no significant differences in terms of clinical scoring. In addition, the mean aMPV vaccine viral titers and antibody titers measured in the dual vaccinated groups were comparable or even higher than in the group vaccinated solely against aMPV. Lastly, based on the NDV viral and antibody titers, the combined aMPV and NDV vaccination does not seem to interfere with protection against NDV, although further studies involving an actual ND challenge will be necessary to fully demonstrate this hypothesis

Development of an empathy and clarity rating scale to measure the effect of medical improv on end-of-first-year OCSE performance: a pilot study

Patients want empathetic physicians who listen and understand. How do you teach and measure empathy? Medical educators, including those inspired by Alan Alda, have turned to theater to teach skills in empathetic communication. Improvisation-informedcurriculum (medical improv) draws upon foundational actors training: deep listening, emotional understanding, connections, authenticity. Arating scale to measure the impact of medical improv on empathetic and clear communication does not exist. Objective: To develop aframework and instrument, the Empathy and Clarity Rating Scale (ECRS), for measuring communication elements used by actors and physicians, and pilot ECRS to test effectiveness of medical improv on first-yearstudents' communication skills. Design: Four medical schools collaborated. USMLE Step 2 Communication and Interpersonal Skills (CIS) domains were used as framework for discussion among three focus groups, each with clinicians, actors, communication experts, and community members with patient experience. Audiotaped discussions were transcribed; open coding procedures located emerging themes. The initial coding scheme was compared with the Consultation and Relational Empathy (CARE) measure. ECRS content was aligned with CARE, CIS and focus group themes. Modified nominal processes were conducted to finalize the scale. We implemented procedures to establish content validity and interrater reliability. Final ECRS was used to study student performance across three levels of experience with medical improv. Results: The final ECRS was comprised of seven five-pointscale items. Narrative comments precede behaviorally anchored ratings: 5=desired, 1=ineffective, 2-4=developing based upon adjustment needed. Rater agreement across all items was 84%. There was asmall correlation between the ECRS and another measure interviewing (r=0.262, p=0.003). Students with advanced medical improv training outperformed those without (F=3.51, p=.042). Conclusion: Acommunication scale enlightened by experiences of actors, clinicians, scholars and patients has been developed. The ECRS has potential to detect the impact of medical improv on development of empathetic and clear communication

Nuove varianti di Bronchite Infettiva negli allevamenti avicoli siciliani

Avian Infectious Bronchitis (IB) is a disease caused by a Coronavirus, included in Coronaviridae family. The disease, endemic in Italy, affects both broilers and laying hens. It represents one of the main health issue in sicilian poultry farms. The presence of new antigenic variants makes problematic the implementation of an adequate prophylaxis through the use of appropriate vaccines. The present work aims to study the spread of IB strains in Sicily by serological and biomolecular tests in order to investigate the presence of "historical" strains as well as new strains and to carry out the genotyping of viruses isolated. The serological results show that the used vaccination protocols are able to develop an adequate antibody titre along all production steps both laying hens and broilers. The virological results underline the presence of QX strain in a broilers farm. This is a strain widespread in Italy but never reported in the regional territory

Valutazione di alcune performance diagnostiche di kit ELISA per la diagnosi sierologica di Anemia Infettiva Equina (AIE)

Data on evaluation of some diagnostic parameters of all ELISA kits available in Italy for the serodiagnosis of AIE are presented and discussed. Ten laboratories were involved using a panel of 30 sera ran with 4 commercial kits and 2 in-house kits. Kits were also evaluated using a panel of sera from vaccinated animals at different days post vaccination (p.v.). All sera were also tested in agar gel immunodiffusion (AGID). The parameters evaluated were: diagnostic sensitivity (DSe) and specificity (DSp), Cohen K, weighted Cohen K, coefficient of variation (CV), accordance, concordance. Analysis of these parameters indicates that all kits have a higher sensitivity than AGID, even if a complete evaluation, as indicated by OIE (1) should be carried out

Creative group performances to assess core competencies in a first-year patient-centered medicine course

Introduction: A novel assessment of systems-based practice and practice-based learning and improvement learning objectives, implemented in a first-year patient-centered medicine course, is qualitatively described. Methods: Student learning communities were asked to creatively demonstrate a problem and solution for health care delivery. Skits, filmed performances, plays, and documentaries were chosen by the students. Video recordings were reviewed for themes and the presence of course competencies. Results: All performances demonstrated not only the index competencies of team work and facilitation of the learning of others, but many other core objectives of the course. The assignment was rated positively both by the faculty and the students, and has been added to the assessment modalities of the course

Limited Susceptibility of Chickens, Turkeys, and Mice to Pandemic (H1N1) 2009 Virus

To determine susceptibility of chickens, turkeys, and mice to pandemic (H1N1) 2009 virus, we conducted contact exposure and inoculation experiments. We demonstrated that chickens were refractory to infection. However, oculo-oronasally inoculated turkeys and intranasally inoculated mice seroconverted without clinical signs of infection

Pathogenicity of Highly Pathogenic Avian Influenza Virus (H5N1) in Adult Mute Swans

Adult, healthy mute swans were experimentally infected with highly pathogenic avian influenza virus A/Cygnus cygnus/Germany/R65/2006 subtype H5N1. Immunologically naive birds died, whereas animals with preexisting, naturally acquired avian influenza virus–specific antibodies became infected asymptomatically and shed virus. Adult mute swans are highly susceptible, excrete virus, and can be clinically protected by preexposure immunity

Risk stratification for arrhythmic death in an emergency department cohort: a new method of nonlinear PD2i analysis of the ECG

Heart rate variability (HRV) reflects both cardiac autonomic function and risk of sudden arrhythmic death (AD). Indices of HRV based on linear stochastic models are independent risk factors for AD in postmyocardial infarction (MI) cohorts. Indices based on nonlinear deterministic models have a higher sensitivity and specificity for predicting AD in retrospective data. A new nonlinear deterministic model, the automated Point Correlation Dimension (PD2i), was prospectively evaluated for prediction of AD. Patients were enrolled (N = 918) in 6 emergency departments (EDs) upon presentation with chest pain and being determined to be at risk of acute MI (AMI) >7%. Brief digital ECGs (>1000 heartbeats, ∼15 min) were recorded and automated PD2i results obtained. Out-of-hospital AD was determined by modified Hinkle-Thaler criteria. All-cause mortality at 1 year was 6.2%, with 3.5% being ADs. Of the AD fatalities, 34% were without previous history of MI or diagnosis of AMI. The PD2i prediction of AD had sensitivity = 96%, specificity = 85%, negative predictive value = 99%, and relative risk >24.2 (p ≤ 0.001). HRV analysis by the time-dependent nonlinear PD2i algorithm can accurately predict risk of AD in an ED cohort and may have both life-saving and resource-saving implications for individual risk assessment

Chances and Limitations of Wild Bird Monitoring for the Avian Influenza Virus H5N1 — Detection of Pathogens Highly Mobile in Time and Space

Highly pathogenic influenza virus (HPAIV) H5N1 proved to be remarkably mobile in migratory bird populations where it has led to extensive outbreaks for which the true number of affected birds usually cannot be determined. For the evaluation of avian influenza monitoring and HPAIV early warning systems, we propose a time-series analysis that includes the estimation of confidence intervals for (i) the prevalence in outbreak situations or (ii) in the apparent absence of disease in time intervals for specified regional units. For the German outbreak regions in 2006 and 2007, the upper 95% confidence limit allowed the detection of prevalences below 1% only for certain time intervals. Although more than 25,000 birds were sampled in Germany per year, the upper 95% confidence limit did not fall below 5% in the outbreak regions for most of the time. The proposed analysis can be used to monitor water bodies and high risk areas, also as part of an early-warning system. Chances for an improved targeting of the monitoring system as part of a risk-based approach are discussed with the perspective of reducing sample sizes