Protection of chickens after live and inactivated virus vaccination against challenge with nephropathogenic infectious bronchitis virus PA/Wolgemuth/98

Protection provided by live and inactivated virus vaccination against challenge with the virulent nephropathogenic infectious bronchitis virus (NIBV) strain PA/Wolgemuth/98 was assessed. Vaccinations with combinations of live attenuated strains Massachusetts (Mass) + Connecticut (Conn) or Mass + Arkansas (Ark) were given by eyedrop to 2-wk-old specific-pathogen-free leghorn chickens. After live infectious bronchitis virus (IBV) vaccination, some chickens at 6 wk of age received an injection of either an oil emulsion vaccine containing inactivated IBV strains Mass + Ark or an autogenous vaccine prepared from NIBV PA/Wolgemuth/98. Challenge with PA/Wolgemuth/98 was given via eyedrop at 10 wk of age. Serum IBV enzyme-linked immunosorbent assay antibody geometric mean titers (GMTs) after vaccination with the combinations of live attenuated strains were low, ranging from 184 to 1,354, prior to NIBV challenge at 10 wk of age. Both inactivated vaccines induced an anamnestic response of similar magnitudes with serum GMTs of 6,232-12,241. Assessment of protection following NIBV challenge was based on several criteria virus reisolation from trachea and kidney and renal microscopic pathology and IBV-specific antigen immunohistochemistry (IHC). Live attenuated virus vaccination alone with combinations of strains Mass + Conn or Mass + Ark did not protect the respiratory tract and kidney of chickens after PA/Wolgemuth/98 challenge. Chickens given a live combination vaccination of Mass + Conn and boosted with an inactivated Mass + Ark vaccine were also susceptible to NIBV challenge on the basis of virus isolation from trachea and kidney butshowed protection on the basis of renal microscopic pathology and IHC. Live IBV-primed chickens vaccinated with an autogenous inactivated PA/Wolgemuth/98 vaccine had the highest protection against homologous virulent NIBV challenge on the basis of virus isolation.     

鸡传染性支气管炎病毒CK/CH/LHLJ/04V的分离鉴定及致弱的初步研究

从黑龙江省某鸡场发病鸡群的肾脏中分离到一株病毒,通过病毒致SPF鸡胚病变特征、对鸡外周血红细胞凝集特性、病毒粒子形态学特征以及RT-PCR鉴定等方面的研究,表明该病毒为鸡传染性支气管炎病毒,并命名为CK/CH/LHLJ/04V。通过对该病毒基因型分析以及对SPF鸡致病性试验发现该病毒为我国近年来流行的一类重要IBV的代表。将该毒株在SPF鸡胚连续传代110代(P110),取不同代次毒进行动物实验。结果显示,该毒株对SPF雏鸡的致病力随鸡胚传代次数的增加而逐渐降低。P110代毒以105.0 EID50/0.1 mL的剂量通过点眼滴鼻接种15日龄SPF雏鸡,鸡群发病率和死亡率均为0。不同代次毒接种SPF鸡对同源毒株P3代强毒的攻击均具有100%保护性。实验表明,IBV毒株CK/CH/LHLJ/04V P110对SPF雏鸡已无致病性,但仍具有良好的免疫原性,可作为研制IB弱毒疫苗的候选毒株。     

Evaluation of inactivated vaccine of the variant 2 (IS-1494 /GI-23) genotype of avian infectious bronchitis

The infectious bronchitis virus (IBV) is the cause of avian infectious bronchitis (IB). IB is one of the most highly contagious diseases, which results in many economic losses in the poultry industry worldwide. The nature of this virus is such that it generates new genotypes continuously. Proper vaccination is the most suitable way of combatting IB. One of the novel genotypes of IBV, which has been circulating in the Middle Eastern countries, is the variant 2 (IS-1494/GI-23) genotype. This study aims to design and produce an autogenous variant 2 vaccines. After isolation and characterization of the Iranian variant 2, the inactivated vaccine was formulated according to the OIE guidelines, and its different aspects (Purity, titration, inactivation, immunization) were evaluated. The designed vaccine passed all of OIE quality control standards. In the assessment process, the protection rate in the groups receiving the variant 2 and commercial vaccines was 67 % and 60 %, respectively. Although the differences were not significant, they indicated better protection, and the viral load in the feces and the kidney of the group receiving the variant 2 vaccine was lower than that in the commercial vaccine. It is suggested that the variant2 strain should be added as one of the local strains to the commercial inactivated vaccines in areas affected by this genotype. The use of this vaccine in layer and breeder flocks can help to protect them against variant 2 during the production phase. Also, the transfer of maternal antibodies to offspring can provide strain-specific immunity for one-day-old chickens.     

Immunogenicity and efficacy of a bivalent vaccine against infectious bronchitis virus

Infectious bronchitis (IB) is a highly contagious viral disease and is responsible for considerable economic losses in the poultry industry, worldwide. To mitigate the IB-associated losses, multiple vaccines are being applied in the sector with variable successes and thus necessitating the development of a potent vaccine to protect against the IB in the poultry. In the present study, we investigated a bivalent live attenuated vaccine consisting of IB virus (IBV) strain H120 (GI-1 lineage) and D274 (GI-12 lineage) to evaluate its protection against heterologous variant of IBV (GI-23 lineage) in chicken. Protection efficacy was evaluated based on the serology, clinical signs, survival rates, tracheal and kidney histopathology and the viral shedding. Results demonstrated that administering live H120 and D274 (named here Classivar®) vaccine in one day-old and 14 days-old provided 100 % protection. We observed a significant increase in the mean antibody titers, reduced virus shedding, and ameliorated histopathology lesions compared to routinely used vaccination regimes. These results revealed that usage of different IBV vaccines combination can successfully ameliorate the clinical outcome and pathology in vaccinated chicks especially after booster vaccination regime using Classivar®. In conclusions, our data indicate that Classivar® vaccine is safe in chicks and may serve as an effective vaccine against the threat posed by commonly circulating IBV strains in the poultry industry.     

Protection induced by infectious laryngotracheitis virus vaccines alone and combined with Newcastle disease virus and/or infectious bronchitis virus vaccines

Two types of live attenuated vaccines have been used worldwide for the control of infectious laryngotracheitis virus (ILTV): 1) chicken embryo origin (CEO) vaccines; and 2) tissue culture origin vaccines (TCO). However, the disease persists in spite of extensive use of vaccination, particularly in areas of intense broiler production. Among the factors that may influence the efficiency of ILTV live attenuated vaccines is a possible interference of Newcastle Disease virus (NDV) and infectious bronchitis virus (IBV) vaccines with the protection induced by ILTV vaccines. The protection induced by CEO and TCO vaccines was evaluated when administered at 14 days of age alone or in combination with the B1 type strain of NDV (B1) and/or the Arkansas (ARK) and Massachusetts (MASS) serotypes of IBV vaccines. Two weeks after vaccination (28 days of age), the chickens were challenged with a virulent ILTV field strain (63140 isolate, group V genotype). Protection was evaluated at 5 and 7 days postchallenge by scoring clinical signs and quantifying the challenge virus load in the trachea using real-time PCR (qPCR). In addition, the viral load of the vaccine viruses (ILTV, NDV, and IBV) was quantified 3 and 5 days postvaccination also using qPCR. The results of this study indicate that the NDV (B1) and IBV (ARK) vaccines and a multivalent vaccine constituted by NDV (B1) and IBV (ARK and MASS) did not interfere with the protection induced by the CEO ILTV vaccine. However, the NDV (BI) and the multivalent (B1/MASS/ARK) vaccines interfered with the protection induced by the TCO vaccine (P < 0.05). Either in combination or by themselves, the NDV and IBV vaccines decreased the tracheal replication of the TCO vaccine and the protection induced by this vaccine, since the ILTV-vaccinated and -challenged chickens displayed significantly more severe clinical signs and ILTV load (P < 0.05) than chickens vaccinated with the TCO vaccine alone. Although NDV and IBV challenges were not performed, the antibody responses elicited by NDV and/or the IBV vaccinations were significantly reduced (P < 0.05) when applied in combination with the CEO vaccine.     

Attenuation, safety, and efficacy of an infectious bronchitis virus GA98 serotype vaccine

In 1998, novel strains of infectious bronchitis virus (IBV) were identified in chickens from the southeastern United States and classified as a new serotype designated Georgia 98 (GA98). Because of the widespread nature of the GA98 virus in the southeastern United States and the lack of adequate protection with the DE072 vaccine, we developed a specific vaccine for the GA98 serotype. The GA98/0470/98 isolate of IBV was passaged in embryonating chicken eggs 70 times, and attenuation of the virus was determined in specific-pathogen-free chicks. Pass 70 of the GA98/0470/98 strain of IBV when given at 1 day of age by coarse spray and at 14 days of age in the drinking water at 1 x 10(4.5) 50% embryo infectious dose/bird protected against the homologous GA98 challenge as well as provided good protection against the DE072-type virus. In addition, the vaccine was shown to be adequately attenuated and safe at a 10x dosage.     

IBV肾型地方分离株弱毒疫苗与同类冻干疫苗的免疫效力试验比较

本试验对人工致弱IBV肾型地方分离株(SF株)与相应进口弱毒疫苗和国产弱毒疫苗进行了免疫效力比较试验。结果表明SF株免疫后产生抗体的时间和抗体效价高低与其他疫苗相当甚至略高,对攻毒鸡的保护率均为100%。剖检实验鸡,采集肾脏样本做肾脏组织切片,观察发现接种致弱的SF株疫苗后,引起的肾脏组织病理变化比选用的2种国产疫苗病理变化轻微,而与进口疫苗引起的病理变化程度相当,属可逆性病变。     

Evaluation of the effectiveness of two infectious bronchitis virus vaccine programs for preventing disease caused by a California IBV field isolate

Infectious bronchitis virus CA99 serotype was isolated from several broiler flocks in Northern California. The virus caused late-onset respiratory disease and increased airsacculitis condemnation in affected flocks despite the use of an established infectious bronchitis virus vaccination program. An experimental study compared Holland/Arkansas and Massachusetts/Arkansas vaccination protocols to determine the efficacy of commercial infectious bronchitis virus vaccines in reducing respiratory disease and airsacculitis lesions found at processing that were associated with a CA99 field isolate. All vaccination groups were given Massachusetts/Connecticut strains of infectious bronchitis virus vaccines at age 1 day followed by vaccination with either Holland/ Arkansas or Massachusetts/Arkansas vaccine strains at 18 days of age. Birds were challenged at age 31 days with a CA99 field isolate. Gross pathology, histopathology, and virus isolation were evaluated. Chickens vaccinated with Holland/Arkansas had marginally better protection against CA99 challenge than chickens vaccinated with Massachusetts/Arkansas, although differences were not statistically significant.     

Vaccine efficacy against Ontario isolates of infectious bronchitis virus

Infectious bronchitis (IB) is an economically important viral disease with worldwide distribution. Every country with an intensive poultry industry has infectious bronchitis virus (IBV). The virus rapidly spreads from bird to bird through horizontal transmission by aerosol or ingestion. Sentinel bird studies were carried out in southern Ontario and IBV has been isolated from layer flocks. Genetic analysis of the S1 region of the strains showed that they were not vaccine related. The pathogenicity of selected Ontario variants of IBV isolates was studied and the subsequent work was to determine the degree of protection against field isolates provided by a commonly used vaccine MILDVAC-Ma5 in Ontario. The protection was evaluated by challenging immunized chickens with the respiratory (IBV-ON1) and nephropathogenic (IBV-ON4) viruses. The mean vaccine efficacy for IBV-ON1 was 66.7% indicating that a Massachusetts serotype vaccine would provide some protection against IBV field isolates.     

两株鸡传染性支气管炎病毒的分离鉴定及S1基因序列分析

为调查广东地区鸡传染性支气管炎病毒(IBV)的流行及其遗传变异情况,本研究通过病料SPF鸡胚接种和鸡胚尿囊液的RT-PCR鉴定,于2013年从广东湛江地区不同发病鸡场分离到两株IBV,分别命名为CK/CH/GD/ZJ10/2013和CK/CH/GD/ZJ11/2013,并对这两株IBV的S1基因进行序列分析。结果显示,CK/CH/GD/ZJ10/2013株S1基因全长1 626 bp,编码542个氨基酸,其裂解位点为NRFRR,属于基因型Ⅲ,并推测其为基因型Ⅲ毒株(CK/CH/GX/NN11-3)与基因型Ⅰ毒株(GX-NN-6)在S1基因处发生重组而产生的新毒株;CK/CH/GD/ZJ11/2013株S1基因全长1620 bp,编码540个氨基酸,其裂解位点为HRRRR,属于基因型Ⅰ(类QX型);两株IBV的S1基因间核苷酸序列及其推导的氨基酸序列同源性较低,与位于同一基因型的参考毒株间同源性较高,而与中国使用的Mass型常规疫苗H120和H52之间的同源性最低,仅为75.7%~76.3%和77.1%~77.9%。本研究可为广东省IBV的流行病学调查和分子生物学研究提供参考。     

Isolation,Identification and Sequence Analysis of S1 Gene of Two Strains of Avian Infectious Bronchitis Viruses

In order to investigate the epidemiology and genetic variation of avian infectious bronchitis virus (IBV) in Guangdong province, two strains of IBV were isolated by inoculation of embryo and RT-PCR detection from diseased chickens at different farms in Zhanjiang, Guangdong province in 2013.We denoted these two strains of IBV as CK/CH/GD/ZJ10/2013 and CK/CH/GD/ZJ11/2013, respectively.Analysis of the S1 gene sequences from these two isolated strains showed that the S1 gene of CK/CH/GD/ZJ10/2013 was 1 626 bp, which encoded 542 amino acids with a cleavage site sequence of NRFRR, while the S1 gene of CK/CH/GD/ZJ11/2013 was 1620 bp, encoded 540 amino acids with a cleavage site sequence of HRRRR.Phylogenetic analysis revealed that these two isolated strains were clustered into genetic groups Ⅲ and Ⅰ(QX-like type), respectively.Homology analysis demonstrated that nucleotide and deduced amino acid sequence homologies between these two isolated strains were lower, while the homologies were higher among the same genotypes, however, the homologies were lower comparing with current vaccine strains such as H120 and H52, with which the nucleotide homologies ranged from 75.7% to 76.3% and the amino acid homologies ranged from 77.1% to 77.9%.Further analysis showed that the CK/CH/GD/ZJ10/2013 strain was formed from a recombination event between CK/CH/GX/NN11-3 and GX-NN-6.Taken together, this study provided valuable insight into prevention and control of IBV infection in Guangdong province.     

传染性支气管炎病毒的分子生物学特性及其弱毒疫苗的研究进展

传染性支气管炎是一种急性、高度接触性传染病,被世界动物卫生组织(OIE)列为B类疫病,对世界养禽业的发展造成了严重威胁.目前,传染性支气管炎的主要预防措施仍然是接种疫苗,但由于传染性支气管炎病毒具有血清型众多、变异速度快且交叉保护性弱等特点,使得传染性支气管炎的防制一直是养禽业面临的重大难题.文章主要对传染性支气管炎病毒的分子生物学特征及其弱毒疫苗的发展历程和免疫策略等方面进行了阐述,以期为新型传染性支气管炎弱毒疫苗的开发及应用提供科学参考.     

Research Progress on Molecular Biological Characteristics and Attenuated Vaccine of Infectious Bronchitis Virus

Infectious bronchitis (IB) is an acute and susceptible infectious disease,which has been classified as B loemia and causes a grave threat to the poultry industry.Now,the primary prevention measures of IB are vaccine inoculation.With the diversity of infectious bronchitis virus (IBV) serotype and easy variation,and the weakness of cross protection,the prevention and control of IB were a major problem in poultry industry.In this ariticle,it summarized the molecular biological characteristics, and the development and immunization strategy of the attenuated vaccine of IB to provide scientific references for the research and application of IB attenuated vaccine.     

Evaluation of the hemagglutination-inhibition test for measuring the response of chickens to avian infectious bronchitis virus vaccination

An infectious bronchitis virus (IBV) hemagglutination-inhibition (HI) test was used to assay serum-antibody titers after IBV vaccination of IBV-susceptible specific-pathogen-free broilers and commercial layers. Three-week-old broilers were vaccinated via eye-drop with IBV strains that represent the antigenic spectrum of commercial vaccines--Holland, Massachusetts 41 (41 Ms), Connecticut 46, Florida 18288, or JMK strain--and revaccinated 3 weeks later with either the same or a heterologous strain. Weekly serum samples were tested by IBV HI with homologous and heterologous antigens. Vaccinates, except for those vaccinated with the Holland strain, were HI-positive with homologous but not heterologous antigens by 1 to 2 weeks postvaccination. Sixteen-week-old IBV-vaccinated commercial layers were revaccinated with IBV Holland 52 (H 52) strain and subsequently infected with Arkansas 99 (Ark 99) and SE 17 strains. In contrast to the limited HI cross-reactivity of serum from IBV-vaccinated broilers, there were extensive cross-reactions in HI tests with 41 Ms, H 52, Ark 99, and SE 17 antigens of revaccinated layers. These results demonstrate that the IBV HI test is more strain-specific than previous reports indicate, especially when the test samples are from early postvaccination.     

A comparative evaluation of the protective efficacy of rMd5deltaMeq and CVI988/ Rispens against a vv+ strain of Marek's disease virus infection in a series of recombinant congenic strains of White Leghorn chickens

Marek's disease (MD) is a lymphoproliferative disease of domestic chickens caused by a highly infectious, oncogenic alpha-herpesvirus known as Marek's disease virus (MDV). MD is presently controlled by vaccination. Current MD vaccines include attenuated serotype 1 strains (e.g., CVI988/Rispens), avirulent serotype 2 (SB-1), and serotype 3 (HVT) MDV strains. In addition, recombinant MDV strains have been developed as potential new and more efficient vaccines to sustain the success of MD control in poultry. One of the candidate recombinant MDV strains, named rMd5deltaMeq, was derived from Md5, a very virulent strain of MDV lacking the MDV oncogene Meq. Our earlier reports suggest that rMd5deltaMeq provided protection equally well or better than commonly used MD vaccines in experimental and commercial lines of chickens challenged with very virulent plus (vv+) strains of MDV. In this study, maternal antibody-positive (trial 1) and negative (trial 2) chickens from a series of relatively MD resistant lines were either vaccinated with the rMd5deltaMeq or CVI988/Rispens followed by infection of a vv+ strain of MDV, 648A, passage 10. This report presents experimental evidence that the rMd5deltaMeq protected significantly better than the CVI988/Rispens (P < 0.01) in the relatively resistant experimental lines of chickens challenged with the vv+ strain of MDV. Together with early reports, the rMd5deltaMeq appeared to provide better protection, comparing with the most efficacious commercially available vaccine, CVI988/Rispens, for control of MD in lines of chickens regardless of their genetic background.     

Genetic diversity of avian infectious bronchitis coronavirus in recent years in China

Fifty-six isolates of avian infectious bronchitis virus (IBV) were obtained from different field outbreaks in China in 2010, and they were genotyped by comparison with 19 reference strains in the present study. The results showed that LX4-type isolates are still the predominant IBVs circulating in chicken flocks in China, and these isolates could be grouped further into two clusters. Viruses in each cluster had favored amino acid residues at different positions in the S1 subunit of the spike protein. In addition, a recombination event was observed to have occurred between LX4- and tl/CH/LDT3/03I-type strains, which contributed to the emergence of a new strain. The most important finding of the study is the isolation and identification of Taiwan II-type (TW II-type) strains of IBV in mainland China in recent years. The genome of TW II-type IBV strains isolated in mainland China has experienced mutations and deletions, as demonstrated by comparison of the entire genome sequence with those of IBV strains isolated in Taiwan. Pathogenicity testing and sequence analysis of the 3' terminal untranslated region revealed that TW II-type IBV strains isolated in mainland China have a close relationship with the embryo-passaged, attenuated TW2296/95.