Additional file 1: of Shedding of Salmonella Typhimurium in vaccinated and unvaccinated hens during early lay in field conditions: a randomised controlled trial

Table S1. List of Salmonella serovars tested for the specificity of wild type Salmonella Typhimurium PCR. (DOCX 15 kb

<i>Salmonella</i><i>typhimurium</i> in the Australian egg industry: Multidisciplinary approach to addressing the public health challenge and future directions

<p>In Australia, numerous egg-related human <i>Salmonella</i> <i>typhimurium</i> outbreaks have prompted significant interest among public health authorities and the egg industry to jointly address this human health concern. Nationwide workshops on <i>Salmonella</i> and eggs were conducted in Australia for egg producers and regulatory authorities. State and national regulators represented Primary Production, Communicable Disease Control, Public Health and Food Safety, and Food Standards Australia and New Zealand. All attendees participated in discussions aimed at evaluating current evidence-based information, issues related to quality of egg production, and how to ensure safe eggs in the supply chain, identifying research gaps and practical recommendations. The perceptions from egg producers and regulatory authorities from various states were recorded during the workshops. We presented the issues discussed during the workshops, including <i>Salmonella</i> in the farm environment, <i>Salmonella</i> penetration across eggshell, virulence in humans, food/egg handling in the supply chain, and intervention strategies. We also discussed the perceptions from egg producers and regulators. Recommendations placed emphasis on the future research needs, communication between industry and regulatory authorities, and education of food handlers. Communication between regulators and industry is pivotal to control egg-borne <i>S. typhimurium</i> outbreaks, and collaborative efforts are required to design effective and appropriate control strategies.</p

Reaction of tM2e-MBP and M2e-MBP proteins with positive sera in ELISA.

<p>The positive sera are described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108420#pone-0108420-t001" target="_blank">Table 1</a>. Pared T test highlighted the statistically higher reactivity (p = 0.05) of tM2e-MBP compared to M2e-MBP.</p

Two-graph receiver operating curve (TG-ROC) analysis of tM2e-MBP ELISA OD for field (vaccinated or non-vaccinated) and challenged (positive) sera.

<p>The cut-off value was chosen to be at 95% sensitivity and 95% specificity.</p

Boxplot of ODs obtained with tM2e-MBP and M2e-MBP with positive, vaccinated and SPF sera showed in <b>table 2</b>.

<p>The asterisks represent small number of the cases which are far from common ranges of plus and minus standard deviation (±SD).</p

SDS-PAGE analysis of purified tM2e-MBP recombinant protein.

<p>Lane M: Molecular weight markers. Lanes 1 to 7: Fractions of purified recombinant tM2e-MBP protein with molecular weight of 52.9 kD (arrow).</p

Positive serum samples used in the study^*.

<p>* Birds were immunized once (D10), twice (B47, B38, B1, B2, PL80) or three times (A17) with a commercial H5N1 vaccine, at four weekly intervals. Two weeks after the last immunization, birds were challenged with H5N1 strains ^a A/chicken/West Java/Sbg-29/2007 or ^b A/Chicken/West Java/PWT-WIJ/2006. Two weeks after the challenge, sera were collected for assay in haemagglutination inhibition test and M2e and tM2e ELISA.</p><p>Positive serum samples used in the study^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108420#nt101" target="_blank">*</a>}.</p

ANOVA and mean comparison by Tukey's method of ELISA OD values of positive, SPF and sera from vaccinated chicks.

<p>* From each positive, vaccinated and SPF groups, three samples were tested in triplicate in both tM2e-MBP and M2e-MBP ELISAs.</p><p>ANOVA and mean comparison by Tukey's method of ELISA OD values of positive, SPF and sera from vaccinated chicks.</p

Evaluation of tM2e-MBP ELISA using sera from infected (challenged), non-vaccinated, and vaccinated chickens.

<p>ANOVA and mean comparison using Tukey's test highlighted significant results in samples from infected birds compared to negative and vaccinated birds at p = 0.01. ANOVA and Tukey's tests showed statistically higher (p = 0.01) reactivity of infected (positive) compared to vaccinated and (negative) sera.</p

Reaction of tM2e-MBP and M2e-MBP recombinant proteins with positive, vaccinated and SPF sera in ELISA.

<p>Each serum sample run in triplicate at three different times and the mean corrected OD is presented as error bar.</p