Citation: Elfeil, W.K.; Youssef, H.; Sedeek, A.; El-Shemy, A.; Abd-Allah, E.M.; Elkady, M.F.; El_Sayed, E.K.; Bazid, A.-H.I.; Abdallah, M.S. Protective Efficacy of Inactivated H9N2 Vaccine in Turkey Poults under Both Experimental and Field Conditions. Vaccines 2022, 10, 2178. https://doi.org/10.3390/ vaccines10122178 Academic Editor: François Meurens Received: 7 November 2022 Accepted: 13 December 2022 Published: 19 December 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). Article Protective Efficacy of Inactivated H9N2 Vaccine in Turkey Poults under Both Experimental and Field Conditions Wael K. Elfeil 1, * , Hefny Youssef 2 , Ahmed Sedeek 2 , Ahmed El-Shemy 3 , Ehab M. Abd-Allah 4 , Magdy F. Elkady 5 , Eman K. El_Sayed 6 , Abdel-Hamid I. Bazid 7 and Mona S. Abdallah 1 1 Avian and Rabbit Medicine Department, Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt 2 Animal Health Research Institute, Agriculture Research Center, Dokki, Giza 12618, Egypt 3 Parasitology and Animal Disease Department, Veterinary Research Institute, National Research Centre, Dokki, Giza 12622, Egypt 4 Veterinary Hospital, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44511, Egypt 5 Poultry Diseases Department, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt 6 Virology Department, Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt 7 Virology Department, Faculty of Veterinary Medicine, University of Sadat City, Sadat City 32958, Egypt * Correspondence: elfeil@vet.suez.edu.eg Abstract: Low pathogenic avian influenza (LPAI) H9N2 virus is one of the major poultry pathogens associated with severe economic losses in the poultry industry (broiler, layers, breeders, and grand- parents’ flocks), especially in endemic regions including the Middle East, North Africa, and Asian countries. This work is an attempt to evaluate the efficacy of whole inactivated H9N2 vaccine (MEFLUVAC TM H9) in turkey poults kept under laboratory and commercial farm conditions. Here, 10,000 white turkey poults (1-day old) free from maternally derived immunity against H9N2 virus were divided into four groups; G1 involved 10 vaccinated birds kept under biosafety level-3 (BLS-3) as a laboratory vaccinated and challenged group, while G2 had 9970 vaccinated turkeys raised on a commercial farm. Ten of those birds were moved to BLS-3 for daily cloacal and tracheal swabbing to check for the absence of any life-threating disease, before conducting analyses. G3 (10 birds) served as a non-vaccinated challenged control under BSL-3 conditions, while G4 (10 birds) was used as a non-vaccinated and non-challenged control under BSL-3 conditions. Sera were collected on days 7-, 14-, 21-, and 28-post-vaccinations to monitor the humoral immune response using a hemagglutination-inhibition (HI) test. At these same intervals, cloacal and tracheal swabs were also checked for any viral infection. The challenge was conducted 28 days post-vaccination (PV) using AI-H9N2 in BSL-3 by intranasal inoculation of 6-log10 embryo infective dose 50 (EID 50 ). At 3-, 6-, and 10-days post-challenge, oropharyngeal swabs were taken from challenged birds to quantify viral shedding by quantitative polymerase chain reaction (qRT-PCR). The results of this study showed that vaccinated groups (G1/2) developed HI titers of 1.38, 4.38, 5.88, and 7.25 log 2 in G1 vs. 1.2, 3.8, 4.9 and 6.2 log 2 in G2 when measured at 7-, 14-, 21- and 28-days PV, respectively, while undetectable levels were recorded in non-vaccinated groups (G3/4). Birds in G3 showed 90% clinical sickness vs. 10% and 20% in G1/2, respectively, over a 10-day monitoring period following challenge. Vacci- nated birds showed a significant reduction in virus shedding in terms of the number of shedders, amount of shed virus and shedding interval over the non-vaccinated challenged birds. Regarding mortality, all groups did not show any mortality, which confirms that the circulating H9N2 virus still has low pathogenicity and cannot cause mortality. However, the virus may cause up to 90% clinical sickness in non-vaccinated birds vs. 10% and 20% in laboratory- and farm-vaccinated birds, respectively, highlighting the role of the vaccine in limiting clinical sickness cases. In conclusion, under the current trial circumstances, MEFLUVAC TM -H9 provided protective seroconversion titers, significant clinical sickness protection and significant reduction in virus shedding either in laboratory- or farm-vaccinated groups after a single vaccine dose. Keywords: turkey; LPAI H9N2; vaccine efficacy; vaccination; farm vaccination; pathogenicity Vaccines 2022, 10, 2178. https://doi.org/10.3390/vaccines10122178 https://www.mdpi.com/journal/vaccines