Supplemental Material, DS1_VET_10.1177_0300985818767996 - Newcastle Disease Virus Infection in Quail

<p>Supplemental Material, DS1_VET_10.1177_0300985818767996 for Newcastle Disease Virus Infection in Quail by Leonardo Susta, Diego Segovia, Timothy L. Olivier, Kiril M. Dimitrov, Ismaila Shittu, Valerie Marcano, and Patti J. Miller in Veterinary Pathology</p

DNA-vaccine groups and plasmid combinations.

<p>DNA-vaccine groups and plasmid combinations.</p

Characterization and evaluation of the effect of co-delivering chIFNγ with a DNA vaccination system.

<p>Plasmids expressing NDV F and chIFNγ genes were developed and characterized for their use as DNA vaccines and adjuvant, respectively. DF-1 cells were transfected with pTriEX, pTriEX-ZJ1-F and pTriEX- IFNγ. Cell culture supernatants were tested by western blotting for the presence of F protein <b>(A)</b> and chIFNγ <b>(B)</b>, respectively. Eighteen-day-old SPF ECEs were inoculated with TE buffer, pTriEX, pTriEX-ZJ1-F, or pTriEX-ZJ1-F plus pTriEX- IFNγ and boosted 2 weeks after hatched. Two weeks after booster vaccination, birds were challenged with vZJ1. Oropharyngeal <b>(C)</b> and cloacal <b>(D)</b> swab samples were collected 3 days after challenge to measure the amount of challenge virus shed into the environment. Viral titers were determined by quantitative real time reverse transcription polymerase chain reaction (qRRT-PCR). A standard was prepared with a vZJ1 virus stock of know concentration, this was included in every plate and was used to obtain viral titers expressed as EID₅₀/mL. Morbidity <b>(E)</b> and mortality <b>(F)</b> were also evaluated. Viral shedding results were analyzed with One-way ANOVA followed by a multiple comparisons Tukey's test. Differences in morbidity among groups were evaluated using a two-tailed Z test for comparison of sample proportions. Survival curves were analyzed using the Long-Rank test. Statistical difference was considered with a <i>P<0</i>.<i>05</i>. Significant differences are denoted by different letters.</p

Effect of chIFNγ on hatchability and survival after <i>in ovo</i> vaccination with rZJ1*L/IFNγ and challenge with vZJ1.

<p>Effect of chIFNγ on hatchability and survival after <i>in ovo</i> vaccination with rZJ1*L/IFNγ and challenge with vZJ1.</p