El-Moeid Amena Abd, El-Deeb Ayman Hany, Elsaied Marwa Fathy, Soliman Reem Ahamed, El-Safty Mounir Mohamed, Hussein Hussein Aly
Department of Virology, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt.
Central Laboratory for Evaluation of Veterinary Biologics, Abassia, Cairo, Egypt.
Vet World. 2021 Aug;14(8):2131-2141. doi: 10.14202/vetworld.2021.2131-2141. Epub 2021 Aug 20.
Highly pathogenic avian influenza H5N8 virus of clade 2.3.4.4 was newly emerged to Egypt and firstly detected in carcasses of wild birds in November 2016. This study assessed the protection efficacy and virus shedding reduction of three different inactivated avian influenza (AI) H5 (H5N1, H5N2, and H5N3) commercial vaccines against challenge with two newly emerging highly pathogenic AI virus H5N8 Egyptian isolates in specific-pathogen-free (SPF) chicks.
10-day-old SPF chicks (n=260) were divided into 20 groups (n=13). Groups 1-5 were vaccinated through the subcutaneous route (S/C) with 0.5 mL of H5N1 vaccine, Groups 6-10 were vaccinated (S/C) with 0.5 mL of H5N2 vaccine, and Groups 11-15 were vaccinated (S/C) with 0.5 mL of H5N3 vaccine. Positive control groups (16-19) were challenged at 25 and 31 days old (2 and 3 weeks post-vaccination [PV]) using H5N8 clade 2.3.4.4 A/duck/Egypt/F13666A/2017(H5N8) and H5N8 clade 2.3.4.4 A/chicken/Egypt/18FL6/2018(H5N8). Group 20 was left non-vaccinated as a control. All vaccinated groups were divided and challenged with both viruses at 25 and 31 days of age. The viral challenge dose was 0.1 mL of 10 EID/0.1 mL titer/chick, and it was administered oronasally. All chicks were kept in isolators for 14 days after each challenge. Sera samples were collected weekly and at 2 weeks post-challenge (PC) to detect a humoral immune response. PC mortalities were recorded daily for 10 days to calculate the protection percentages. Tracheal swabs were collected from the challenged chicks in different groups at 3, 5, 7, and 10 days PC. Kidneys and spleens were collected at 3, 5, 7, and 10 days PC and kept in formalin for histopathological examination to assess lesions and severity scores. Tracheal swabs were inoculated in 10-day-old SPF embryonated chicken eggs for virus titration and to calculate shedding levels.
All studied vaccines displayed 70-100% protection within 10 days PC. Hemagglutination inhibition results from sera samples revealed antibody titers ranging from 0.6 to 5.4 log starting at 1-week PV with the highest titers at 4 weeks PV. Challenged SPF chickens exhibited a notable reduction in virus shedding, with an average of 1.5-2 log, compared to control birds. Various histopathological lesions with different scores were detected.
Our findings suggest that the inadequate virus shedding reduction and protection efficacy of studied vaccines were variable and that the type of vaccine to be used under field conditions should be reconsidered. Study of the variability between the Egyptian old emerged AI (AIV) 2017 H5N8 strains and the new emerging AIV 2018 H5N8 is required to achieve optimal protection and limit the current economic losses.
2.3.4.4分支的高致病性禽流感H5N8病毒新近出现在埃及,并于2016年11月首次在野生鸟类尸体中被检测到。本研究评估了三种不同的灭活禽流感(AI)H5(H5N1、H5N2和H5N3)商业疫苗对无特定病原体(SPF)雏鸡感染两种新出现的高致病性AI病毒H5N8埃及分离株的保护效果及病毒排出量的减少情况。
将10日龄的SPF雏鸡(n = 260)分为20组(n = 13)。第1 - 5组通过皮下途径(S/C)接种0.5 mL H5N1疫苗,第6 - 10组通过皮下途径(S/C)接种0.5 mL H5N2疫苗,第11 - 15组通过皮下途径(S/C)接种0.5 mL H5N3疫苗。阳性对照组(16 - 19组)在25日龄和31日龄(接种疫苗后[PV]2周和3周)使用2.3.4.4分支的H5N8 A/duck/Egypt/F13666A/2017(H5N8)和2.3.4.4分支的H5N8 A/chicken/Egypt/18FL6/2018(H5N8)进行攻毒。第20组不接种作为对照。所有接种疫苗的组在25日龄和31日龄时均分为两组,分别用两种病毒进行攻毒。病毒攻毒剂量为0.1 mL含10 EID/0.1 mL效价/雏鸡,经口鼻腔接种。每次攻毒后,所有雏鸡在隔离器中饲养14天。每周以及攻毒后(PC)2周采集血清样本以检测体液免疫反应。记录攻毒后10天内每天的PC死亡率以计算保护率。在攻毒后3、5、7和10天从不同组的攻毒雏鸡采集气管拭子。在攻毒后3、5、7和10天采集肾脏和脾脏并保存在福尔马林中用于组织病理学检查以评估病变和严重程度评分。将气管拭子接种到10日龄的SPF鸡胚中进行病毒滴定并计算排出水平。
所有研究的疫苗在攻毒后10天内显示出70 - 100%的保护率。血清样本的血凝抑制结果显示,从接种疫苗后1周开始抗体效价范围为0.6至5.4 log,在接种疫苗后4周时效价最高。与对照鸡相比,攻毒的SPF鸡的病毒排出量显著减少,平均减少1.5 - 2 log。检测到具有不同评分的各种组织病理学病变。
我们的研究结果表明,所研究疫苗在减少病毒排出和保护效果方面存在不足且各不相同,应重新考虑在野外条件下使用的疫苗类型。需要研究埃及2017年出现的旧型AI(AIV)H5N8毒株与2018年新出现的AIV H5N8之间的差异,以实现最佳保护并限制当前的经济损失。