Identification of New Mycobacterium bovis antigens and development of a multiplexed serological bead-immunoassay for the diagnosis of bovine tuberculosis in cattle.

Serological assays for bovine tuberculosis diagnosis require the use of multiple Mycobacterium bovis specific antigens to ensure the detection of infected animals. In the present study, identification and selection process of antigens, based on data from published proteomic studies and involving the use of bioinformatics tools and an immuno-screening step, was firstly performed for identifying novel antigens that elicit an antibody response in M. bovis infection.

Performance of two commercial serological assays for bovine tuberculosis using plasma samples.

In the bovine tuberculosis diagnosis, the use of plasma samples (already available for IFNɣ assays) in serological tests might facilitate the work in the field. Here, the performance of two commercial serological tests (ELISA IDEXX M. bovis Ab test and Enferplex Bovine TB antibody test) were evaluated using plasma samples from cattle in Belgium.

Field evaluation of two commercial serological assays for detecting bovine tuberculosis.

Diagnosis of bovine tuberculosis in cattle is challenging due to complex immune host response to infection that limit the performance of available diagnostic tests. In this study, performance of two commercial serological assays developed to detect bovine tuberculosis were evaluated: Enferplex Bovine TB antibody kit including 11 antigens (EnferGroup, Ireland) and IDEXX M. bovis Ab kit (IDEXX, USA).

Domestic canaries (Serinus canaria forma domestica) are susceptible to low pathogenic avian influenza virus infections.

Avian influenza viruses have been isolated from many bird species; however, little is known about the susceptibility of pet birds to low pathogenic avian influenza (LPAI) viruses. To address this research gap, domestic canaries (Serinus canaria forma domestica) were experimentally infected with H5 and H7 LPAI viruses to determine susceptibility and to evaluate samples for diagnostic purposes. Clinical evidence of infection (e.

Specific antibody-mediated immunity in the reproductive tract of laying chickens immunized against Newcastle disease with conventional attenuated and inactivated vaccines.

Despite the widespread and successful use of Newcastle disease (ND) vaccines, Newcastle disease virus (NDV) can seriously injure the reproductive tract of egg-laying hens, leading to rapid egg-drop and poor shell quality. Few published studies investigated local NDV-specific immune response in the reproductive tract after ND vaccination of hens. The present study investigated, for the first time, local NDV-specific antibody-mediated immunity in segments of the oviduct during the laying period.

Impact of Age, Season, and Flowing vs. Stagnant Water Habitat on Avian Influenza Prevalence in Mute Swan (Cygnus olor) in Belgium.

Due to their probable role in the spread of Asian highly pathogenic avian influenza (HPAI) H5N1 virus, and in order to explore its implication in the low pathogenic avian influenza (LPAI) virus epidemiology, mute swans represent one particular wild bird species specifically targeted in the avian influenza (AI) surveillance elaborated in Belgium. A total of 640 individual mute swans have been sampled during a 4-yr AI surveillance program (2007-2010) to determine the AI seroprevalence and viroprevalence in this species; all were analyzed through age, temporal, and habitat (flowing and stagnant water) factors. Using a nucleoprotein (NP)-based ELISA, a global antibody prevalence of 35% has been found and was characterized by two peaks in the winter and the summer that might be indicative of a greater LPAI virus circulation in the autumn than in the spring.

Protection Afforded by a Recombinant Turkey Herpesvirus-H5 Vaccine Against the 2014 European Highly Pathogenic H5N8 Avian Influenza Strain.

A highly pathogenic avian influenza (HPAI) H5N8 (clade 2.3.4.

Complete Coding Sequences of One H9 and Three H7 Low-Pathogenic Influenza Viruses Circulating in Wild Birds in Belgium, 2009 to 2012.

The complete coding sequences of four avian influenza A viruses (two H7N7, one H7N1, and one H9N2) circulating in wild waterfowl in Belgium from 2009 to 2012 were determined using Illumina sequencing. All viral genome segments represent viruses circulating in the Eurasian wild bird population.

First TBEV serological screening in Flemish wild boar.

In the frame of a Flemish wildlife surveillance in 2013, a serological screening was performed on sera from wild boar (Sus scrofa; n=238) in order to detect tick-borne encephalitis virus (TBEV)-specific antibodies. Neutralising antibodies were titrated with a seroneutralisation test (SNT), using two cut-off titres (1/10-1/15). Seven wild boars were found TBEV-seropositive and showed moderate (>1/15) to high (>1/125) SNT-titres; three individuals had borderline results (1/10-1/15).

Evaluation of the kinetics of anti-NP and anti-HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus.

Serological monitoring is a feature of surveillance programmes for the detection of the circulation of notifiable low pathogenic avian influenza (LPAI) viruses in commercial poultry holdings. Commercial multispecies nucleoprotein (NP) enzyme-linked immunosorbent assays (ELISAs) have been replacing the haemagglutination inhibition (HI) test as pre-screening tools. Few comparative studies have been conducted to test sera from domestic ducks for diagnostic purposes.

Multiyear Serological Surveillance of Notifiable Influenza A Viruses in Belgian Poultry: A Retrospective Analysis.

Surveillance of notifiable avian influenza (NAI) virus is mandatory in European member states, and each year a serological survey is performed to detect H5 and H7 circulation in poultry holdings. In Belgium, this serological monitoring is a combination of a stratified and a risk-based approach and is applied to commercial holdings with more than 200 birds. Moreover, a competitive nucleoprotein (NP) ELISA has been used as first screening method since 2010.

Extended transmission of two H5/H7 low pathogenic avian influenza viruses in chickens.

Transmission experiments are useful for investigating the mechanisms of low pathogenic notifiable avian influenza virus (LPNAI) transmission. In this study, the hypothesis that inoculation-infected chickens are more infectious than contact-infected chickens was tested. To this end, extended transmission experiments with one H5N2 and one H7N1 LPAIV which had previously been characterized in a series of standard transmission experiments were conducted in specific pathogen-free (SPF) chickens.

An experimental model to analyse the risk of introduction of a duck-originated H5 low-pathogenic avian influenza virus in poultry through close contact and contaminative transmission.

Aquatic wild birds are often carriers of low-pathogenic avian influenza viruses (LPAIVs). If H5 and H7 LPAIVs are transmitted to poultry and have the opportunity to circulate, a highly pathogenic AIV may arise. Contact with aquatic wild birds is one of the most important ways in which these LPAIVs can be introduced into poultry flocks.

Different replication profiles in specific-pathogen-free chickens of two H7 low pathogenic avian influenza viruses isolated from wild birds.

During an active wild bird survey conducted in Belgium from 2007 to 2011, two low pathogenic avian influenza (LPAI) H7 viruses were isolated from wild birds: an H7N1 virus from a common shelduck (Tadorna tadorna) and an H7N7 virus from a Canada goose (Branta canadensis). The H7 sequence analyses and intravenous pathogenicity indices indicated that they were both low pathogenic isolates and genetically related to other recent European H7 LPAIs isolated from wild birds. Interestingly, the two isolates showed different replication profiles in specific-pathogen-free (SPF) chickens, but poultry can be at risk from both.

Evaluation of four enzyme-linked immunosorbent assays for the serologic survey of avian influenza in wild bird species.

Wild birds that reside in aquatic environments are the major reservoir of avian influenza viruses (AIVs). Since this reservoir of AIVs forms a constant threat for poultry, many countries have engaged in AIV surveillance. More and more commercial enzyme-linked immunosorbent assays (ELISA) are available for serologic surveillance, but these tests are often developed and validated for use in domestic poultry.

The impact of viral tropism and housing conditions on the transmission of three H5/H7 low pathogenic avian influenza viruses in chickens.

In this study, shedding and transmission of three H5/H7 low pathogenic avian influenza viruses (LPAIVs) in poultry was characterized and the impact of floor system on transmission was assessed. Transmission experiments were simultaneously conducted with two groups of animals housed on either a grid or a floor covered with litter. Transmission was observed for H5N2 A/Ch/Belgium/150VB/99 LPAIV.

Chasing notifiable avian influenza in domestic poultry: a case report of low-pathogenic avian influenza h5 viruses in two Belgian holdings.

In December 2008, bird species in two geographically distant holdings were found positive for H5 viruses following the annual Avian influenza serological screening in Belgium. The virological tests performed identified in one holding a low-pathogenic avian influenza (LPAI) virus subtype H5N2, and a H5 LPAI virus was identified by real-time PCR and direct sequencing at the second holding. The first farm was an outdoor mixed holding housing ornamental birds and poultry (n = 6000) and the second a free-range geese breeding farm (n = 1500).

H5N1 high pathogenicity avian influenza virus survival in different types of water.

Persistence of H5N1 high pathogenicity avian influenza virus (HPAIV), isolated during the epidemic in wild birds in Poland in 2006, was evaluated in three water samples derived from the sources known to host wild water birds (city pond, Vistula river mouth, and Baltic Sea). The virus was tested at two concentrations (10(4) and 10(6) median tissue culture infective dose per milliliter) and at three temperatures (4 C, 10 C, and 20 C), representing average seasonal temperatures in Poland. All tested water samples were filtered before virus inoculation, and one unfiltered sample (Baltic seawater) was also tested.

Evaluation of different serologic markers for the early detection of avian influenza infection in chickens.

Viruses of all subtypes may be introduced into domestic poultry, but only H5 and H7 low pathogenicity avian influenza viruses can mutate during circulation in poultry and emerge as high pathogenicity avian influenza variants. It is therefore essential to monitor the field situation continuously to detect low pathogenicity notifiable avian influenza (LPNAI) as soon as possible. With the emergence of the highly pathogenic H5N1, markers of infection of avian influenza are becoming more and more significant.

Evaluation of rapid antigen detection kits for the diagnosis of highly pathogenic avian influenza H5N1 infection.

Early detection of highly pathogenic (HP) strains of avian influenza, especially the HP H5N1, is important in terms of controlling and minimizing the spread of the virus. Several rapid antigen detection kits that are able to detect influenza A viruses in less than 1 hr are commercially available, but only a few of them have been evaluated. In this study, four commercially available rapid tests for veterinary usage and two tests for human usage were evaluated and compared.

Rapid detection of Eurasian and American H7 subtype influenza A viruses using a single TaqManMGB real-time RT-PCR.

A real-time reverse transcription PCR (RRT-PCR) targeting a highly conserved HA2 H7 region was developed for the detection of all H7 subtype avian influenza viruses (PanH7). The wide phylogenetic scope and analytical sensitivity and specificity were validated with the use of a panel of 56 diverse influenza A viruses. The detection limit was determined with the use of serial dilutions of Eurasian isolates A/Ck/BE/06775/2003 and A/Ck/It/1067/v99 and North American isolates A/CK/PA/ 143586/2001 and A/Quail/PA/20304/1998, to be 1 log10 higher than the detection limit of the generic influenza A matrix RRT-PCR (about 2.

Evaluation of different strategies for the use of ELISA tests as first screening tools for serologic surveillance of low pathogenic avian influenza in the Belgian poultry sector.

Since the emergence of the highly pathogenic avian influenza H5N1, avian influenza surveillance has been expanded in Europe. The serologic monitoring of domestic poultry is usually accomplished using the reference hemagglutination inhibition (HI) test for the detection of H5 and H7 subtypes. However, as the number of tested sera has been increasing, there is a need for another serologic method that could be used as a preliminary screening test.

Redesigning the serological surveillance program for notifiable avian influenza in Belgian professional poultry holdings.

This study was aimed at redesigning the Belgian active surveillance program for domestic birds in professional poultry holdings based on a risk analysis approach. A stochastic quantitative analysis, combining all data sources, was run to obtain sensitivity estimates for the detection of an infected bird in the different risk groups identified. An optimal number of holdings for each risk group was then estimated on the basis of the different sensitivities obtained.

Longitudinal analysis of the costs associated with inpatient initiation and subsequent outpatient continuation of proton pump inhibitor therapy for stress ulcer prophylaxis in a large managed care organization.

Background: Proton pump inhibitor (PPI) therapy is commonly initiatedin hospitals for a variety of reasons including stress ulcer prophylaxis. Outpatient use of inpatient-initiated PPI use may be medically unwarranted.

Objective: To (a) describe in a longitudinal analysis the incidence and reasons for hospital initiation of PPI therapy, (b) identify the proportion of members continued on PPI therapy at hospital discharge that is not medically warranted, and (c) estimate the total costs incurred by the managed care organization (MCO) and its members due to inappropriate continuation of hospital-initiated PPI therapy after discharge.