Chicken Infectious Anemia Research Articles

Construction of Recombinant Marek's Disease Virus Co-Expressing VP1 and VP2 of Chicken Infectious Anemia Virus.

The chicken infectious anemia virus (CIAV) has been reported in major poultry-producing countries and poses a significant threat to the poultry industry worldwide. In this study, two Marek's disease virus (MDV) recombinants, rMDV-CIAV-1 and rMDV-CIAV-2, were generated by inserting the CIAV VP1 and VP2 genes into the MDV vaccine strain 814 at the US2 site using the fosmid-based rescue system. For rMDV-CIAV-1, an internal ribosome entry site was inserted between VP1 and VP2, so that both proteins were produced from a single open reading frame. In rMDV-CIAV-2, VP1 and VP2 were cloned into different open reading frames and inserted into the MDV genome. The recombinant viruses simultaneously expressed VP1 and VP2 in infected chicken embryo fibroblasts and exhibited growth kinetics similar to those of the parent MDV. The two recombinant viruses induced antibodies against CIAV in chickens. A single dose of the recombinant viruses provided strong protection against CIAV-induced anemia in chickens. These recombinant VP1- and VP2-expressing MDVs are potential vaccines against CIAV in chickens.

A New Dual Fluorescence Method for Rapid Detection of Infectious Bronchitis Virus at Constant Temperature.

Infectious bronchitis virus (IBV) causes infectious bronchitis in chicken, an acute, highly contagious respiratory infection. Because of genetic mutations and recombination, IBV forms many subtypes, which makes it difficult to treat the disease and apply commercial vaccines. Therefore, to detect IBV in time and stop the virus from spreading, a novel and convenient IBV detection technology based on reverse transcription recombinase-aided amplification (RT-RAA) was established in this study. According to the S1 gene of IBV CH I-V and Mass genotypes and S1 gene of IBV CH VI genotype, a set of optimal primers were designed and selected to establish a real-time dual fluorescence RT-RAA method. The lowest detection line was 10 copies/μL of RNA molecules and the method exhibited no cross-reactivity with avian reticuloendotheliosis virus (REV), infectious bursal disease virus (IBDV), avian leukosis virus (ALV), Newcastle disease virus (NDV), chicken infectious anemia virus (CIAV), infectious laryngotracheitis virus (ILTV), Marek's disease virus (MDV), and H9N2 avian influenza virus (H9N2), demonstrating high specificity. When compared to qPCR detection results, our method achieved a sensitivity of 96.67%, a specificity of 90%, and a Kappa value of 0.87 for the IBV CH I-V and Mass genotypes, and achieved a sensitivity of 100%, a specificity of 97.73%, and a Kappa value of 0.91 for the IBV CH VI genotype.

Synergistic pathogenicity of vertically transmitted chicken infectious anemia virus and avian leukosis virus subgroup J coinfection in chickens

Avian leukemia virus subgroup J (ALV-J) and chicken infectious anemia virus (CIAV) can be vertically transmitted; however, the pathogenicity of vertically transmitted coinfection with these 2 pathogens has not been studied. In this study, we created a model of chick morbidity in which chicks carried either ALV-J, CIAV, or both viruses via embryo inoculation. Thereafter, we analyzed the effects of vertically transmitted coinfection with CIAV and ALV-J on the pathogenicity of ALV-J and performed a purification assay based on hatching, mortality viremia positivity, and detection of fecal ALV-p27 antigen rates, and body weight. The hatching rate of the ALV-J+CIAV group was 68.57%, lower than those of the single infection and control groups. The survival curve showed that the mortality rates of the CIAV and ALV-J coinfection groups were higher than those of the single infection and control groups. Body weight statistics showed that coinfection aggravated the 7-d growth inhibition effect. The results of ALV-p27 antigen detection in cell culture supernatants showed that the positivity rates of the ALV-J and ALV-J+CIAV groups were 100% at all ages and 0% in the control group. The results of ALV-p27 antigen detection by anal swabs showed that the positivity rates of the ALV-J group were 92.86, 90.90, 88.89, and 93.33% at all ages, and that the ALV-J p27 positivity detection rate of anal swabs was lower than that of plasma virus isolation. The immune organ index of the ALV-J+CIAV group was significantly or very significantly lower than those of the single infection and control groups. The immune organ viral load showed that coinfection with CIAV and ALV-J promoted the proliferation of ALV-J and CIAV in immune organs. Coinfection with ALV-J and CIAV reduced chicken embryo hatchability and increased chick mortality and growth inhibition relative to their respective single infections. Additionally, coinfection with ALV-J + CIAV was even more detrimental in inducing immune organ atrophy (e.g., the thymus, spleen, and bursa), and promoted individual virus replication during coinfection.

Synergy of Subgroup J Avian Leukosis Virus and Chicken Infectious Anemia Virus Enhances the Pathogenicity in Chickens.

Subgroup J avian leukemia virus (ALV-J) and chicken infectious anemia virus (CIAV) are widely acknowledged as significant immunosuppressive pathogens that commonly co-infect chickens, causing substantial economic losses in the poultry industry. However, whether co-infection of ALV-J and CIAV have synergistic pathogenicity remains uncertain. To explore their synergistic pathogenesis, we established a co-infection model of ALV-J and CIAV in HD11 cells and specific-pathogen-free (SPF) chickens. We discovered that ALV-J and CIAV can synergistically promote the secretion of IL-6, IL-10, IFN-α, and IFN-γ and apoptosis in HD11 cells. In vivo, compared to the ALV-J and CIAV mono-infected group, the mortality increased significantly by 27% (20 to 47%) and 14% (33 to 47%) in the co-infected group, respectively. We also discovered that ALV-J and CIAV synergistically inhibited weight gain and exhibited more severe organ damage in co-infected chickens. Furthermore, we found that CIAV can promote the replication of ALV-J in HD11 cells and significantly enhance ALV-J viral load in blood and tissues of co-infected chickens, but ALV-J cannot promote the replication of CIAV. Moreover, by measuring the immune organ indexes and proportions of blood CD3+CD4+ and CD3+CD8+ lymphocytes, more serious instances of immunosuppression were observed in ALV-J and CIAV co-infected chickens than in mono-infected chickens. Taken together, our findings demonstrate that ALV-J and CIAV synergistically enhance pathogenicity and immunosuppression.

Isolation and genomic characterization of chicken infectious anemia virus in Jiangsu province of China during 2020-2022.

As an immunosuppressive disease virus, chicken infectious anemia virus (CIAV) mainly infects chickens, causing aplastic anemia and systemic lymphoid tissue atrophy. In recent years, the prevalence of CIAV in the poultry industry globally has caused huge economic losses. In this study, a total of 223 clinical samples, including anal swabs, tissues, blood, and vaccines, were collected from 19 broiler farms or breeding companies in Jiangsu province, with symptoms of significant anemia and immunosuppression during 2020-2022. Among them, 75 samples (75/223, 33.6%) were positive for CIAV in polymerase chain reaction (PCR) test, and 20 CIAV strains were successfully isolated. The phylogenetic trees of the 20 isolates and 42 CIAV strains deposited in GenBank formed four distinct groups (A-D). And the isolates mainly belonged to Group A but with high genetic diversity. Analysis for VP1 indicated that these isolates possess key characteristics of highly pathogenic strains. Meanwhile, VP2 and VP3 were much conserved with much fewer mutations compare to VP1. The above epidemiological study of CIAV provides novel insights into molecular characterization of CIAV and lays the foundation for developing efficient strategies for control of CIAV in China.

First report of co-infections of Marek's disease virus and chicken infectious anaemia virus in poultry flocks in Nigeria

Marek's disease (MD) and chicken infectious anaemia (CIA) are viral immunosuppressive diseases of poultry caused by the MD virus (MDV) and CIA virus (CIAV) respectively. Despite vaccination against MD, the incidence of the disease in vaccinated poultry flocks in Nigeria persists. However, underlying factors like co-infection with CIAV have not been investigated in the country. This study was designed to investigate possible co-infections of MDV and CIAV in poultry flocks in Nigeria. In 2016, tumorous tissue samples were collected from suspected cases of MD at necropsy in Jos, Plateau State, Nigeria. The samples collected were fixed in formalin for histopathological examination, genomic DNA was extracted from a second part and analysed by polymerase chain reaction (PCR), targeting the meq and VP1 genes of the MDV and CIAV, respectively. The histology results revealed that the cutaneous and proventricular lymphomas were characterized by large numbers of mononuclear cellular infiltrates admixed with heterophils. The PCR results revealed that MDV was detected in 66.7% (16/24), CIAV in 45.8% (11/24), and co-infections of MDV and CIAV were detected in 45.8% (11/24) of the samples analysed. In addition, co-infections of MD and CIA were recorded in 100% (6/6) and 27.7% (5/18) of broilers and layer/pullet’ samples respectively. Phylogenetic analysis of the meq gene sequences revealed that the Nigerian MDV clusters with very virulent MDV from Egypt and Italy. While, CIAV sequences were genotype II and genotype III and clustered with CIAVs from Cameroon and China. This is the first report of co-infections of MD and CIA in Nigeria.

Genetic characterization of chicken infectious anaemia viruses isolated in Korea and their pathogenicity in chicks.

Chicken infectious anaemia virus (CIAV) causes severe anemia and immunosuppression through horizontal or vertical transmission in young chickens. Especially, vertical transmission of virus through the egg can lead to significantly economic losses due to the increased mortality in the broiler industry. Here, 28 CIAV complete sequences circulating in Korea were first characterized using the newly designed primers. Phylogenetic analysis based on the complete sequences revealed that CIAV isolates were divided into four groups, IIa (2/28, 7.1%), IIb (9/28, 32.1%), IIIa (8/28, 28.6%) and IIIb (9/28, 32.1%), and exhibited a close relationship to each other. The major groups were IIb, IIIa and IIIb, and no strains were clustered with a vaccine strain available in Korea. Also, for viral titration, we newly developed a quantitative PCR assay that is highly sensitive, reliable and simple. To investigate the pathogenicity of three major genotypes, 18R001(IIb), 08AQ017A(IIIa), and 17AD008(IIIb) isolates were challenged into one-day-old specific-pathogen-free (SPF) chicks. Each CIAV strain caused anaemia, severe growth retardation and immunosuppression in chickens regardless of CIAV genotypes. Notably, a 17AD008 strain showed stable cellular adaptability and higher virus titer in vitro as well as higher pathogenicity in vivo. Taken together, our study provides valuable information to understand molecular characterization, genetic diversity and pathogenicity of CIAV to improve management and control of CIA in poultry farm.

Exploring Seroprevalence Variations of Chicken Infectious Anaemia Virus in Local and Exotic Chickens within Jos, Plateau State, Nigeria

Exploring Seroprevalence Variations of Chicken Infectious Anaemia Virus in Local and Exotic Chickens within Jos, Plateau State, Nigeria

Subclinical circulation of Chicken infectious anaemia virus- A Sero-molecular study.

Chicken infectious anaemia-an important immunosuppressive viral disease of chicken-gained much attention in the recent past. Based on huge mortality and production loss observed in the fast-growing poultry sector, the present study aimed to find out the current status of the chicken infectious anaemia virus (CIAV), among chicken flocks in the Punjab state of India by sero-molecular study. The sera from the blood samples were tested for anti-CIAV antibodies by indirect ELISA and also compared with haematological parameters. DNA from sero-positive samples underwent PCR amplification, sequencing and phylogenetic analysis of the most conserved genomic region (VP3 gene) to detect viraemia in asymptomatic birds. The serological study using indirect ELISA showed a high sero-positivity of 77.27% in chicken flocks. Additionally, the present study also revealed the high molecular evidence (72.54%) of CIAV in apparently healthy birds. Genetic analysis showed that all CIAVs have conserved VP3 genes without any nucleotide substitutions, indicating presence of CIAV and its subclinical circulation among apparently healthy flocks. The wide distribution of CIAV among birds may be the reason for huge mortality and production loss. Further, it is suggested that studies be conducted to find out the co-involvement of CIAV with other immunosuppressive microbial agents and the immunosuppressive effect of CIAV in apparently healthy birds. Also, its role in vaccine failure and outbreaks of various other avian diseases needs to be explored.

Molecular Characteristics of Chicken Infectious Anemia Virus in Central and Eastern China from 2020 to 2022.

To evaluate the recent evolution of CIAV in China, 43 flocks of chickens from the provinces of Henan, Jiangsu, Hubei, and Anhui were screened via polymerase chain reaction during 2020-2022. Of these, 27 flocks tested positive for CIAV nucleic acids, including 12 which were positive for other immunosuppression viruses. Additionally, 27 CIAV strains were isolated, and their whole genomes were sequenced. The AH2001 and JS2002 strains shared the highest identity at 99.56%, and the HB2102 and HB2101 strains shared the lowest identity at 95.34%. Based on the genome sequences of these strains and reference strains, a phylogenetic tree was constructed and divided into eight main branches. Most of the strains were grouped with the East Asian strains, whereas the HB2101 strain belonged to the Brazil and Argentina cluster. A recombination event was detected in multiple strains, in which AH2002 recombined from KJ728827/China/2014 (from Taiwan Province) and HN2203, and AH2202 recombined from KX811526/China/2017 (from Shandong Province) and HN2203. All the obtained strains had a highly pathogenic Gln amino acid site at position 394 of the VP1. Overall, our findings demonstrate the importance of CIAV monitoring and provide data that aid in understanding the evolution of CIAV.

Detection and Molecular Characterization of VP1 Gene of Chicken Infectious Anaemia Virus in Suspected Layer Birds

Background: Chicken Infectious Anaemia Virus (CIAV) is one of the emerging pathogen of the poultry. The Genus of this virus is Gyrovirus and the family Anelloviridae. Viral DNA contains three open reading frames (ORFs) encoding for three proteins VP1, VP2 and VP3. VP1 protein is major structural protein and is a capsid protein having major role in growth and spread of the virus. Present study was aimed for molecular detection of VP1 gene of the CIA virus in the samples collected from the outbreaks occurred at four different district of Maharashtra. Methods: Total 50 samples comprising of Spleen, bone marrow, thymus and liver, were collected. DNA extracted from those samples was used for PCR amplification targeting, VP1 gene. Selected positive purified PCR products of VP1 were subjected for Nucleotide sequencing and Phylogenetic analysis. Results: Total out of 50 pooled samples, 10 samples were positive for VP1 gene i.e. 20 %. Sequencing of coding region of four CAV positive samples was conducted. Genetic analysis showed three isolates had high similarity with Del-Ros vaccine strainfrom USA ; while one isolate shared close similarity to attenuated cloned isolate 10 of Cux-1 strain. The data also indicated Q139 and Q144 and Q394 amino acids substitutions among the VP1. Phylogenetic analysis of the sequenced viruses based on either the coding nucleotide sequence or VP1 coding sequence, suggested the circulation of III genotypes in Maharashtra.

Isolation, Identification, and Whole Genome Analysis of Chicken Infectious Anemia Virus in an Outbreak of Disease in Adult Layer Hens.

Chicken infectious anemia (CIA) poses a significant threat to the chicken industry in China. Due to its non-specific symptoms, the disease is often overlooked. This study aimed to conduct a comprehensive analysis of the etiology and pathology of CIA in Guangxi Province, China. Three strains of the chicken infectious anemia virus (CIAV) were isolated from liver samples of diseased 20-week-old chickens. The complete genomes of these strains were sequenced, and experiments on specific pathogen-free (SPF) chicks revealed that the GX21121 strain exhibited high virulence. Histopathological examination of the deceased chickens showed liver cell necrosis, fibrous serous exudation, inflammatory cell infiltration, hemorrhage in liver tissues, and congestion in lung and renal tissues. Phylogenetic analysis of the genome revealed that the three strains had a close genetic relationship to the Heilongjiang wild-type strain (GenBank KY486144). The genetic evolution of their VP1 genes indicated that all three CIAV isolates belonged to genotype IIIc. In summary, this study demonstrated the genomic diversity of three CIAV strains in adult layer hens. The isolation and characterization of the GX21121 strain as a highly virulent isolate provide valuable information for further investigations into the etiology, molecular epidemiology, and viral evolution of CIAV.

Effects of garlic, onion, and apple cider vinegar as a herbal mixture on performance and blood traits of broilers inoculated with chicken infectious anemia virus

This study assessed the effects of a herbal mixture (HM) to protect poultry against chicken infectious anemia (CIA) and to modulate the adverse effects of this virus on performance, mortality, blood profile, white blood cells (WBCs) count, liver enzymes, liver histopathology, and intestinal morphology. Therefore, 240 one-day-old male broiler chicks (Ross 308) were divided into four experimental groups, with six replicates and ten chicks per group. The experimental groups consisted of a control group and groups with 2.5%, 5%, and 7.5% HM, all based on corn-soybean meal. All chicks were inoculated with the CIA virus (CIAV) on day 7. The results showed that supplementation of 2.5% of HM to broiler diet increased feed intake (FI) (P < 0.05) and also increased body weight (BW) and weight gain (WG) slightly (P > 0.05). Adding 7.5% HM caused a reversible decrease in FI, BW, and WG and increased FCR. Compared with the control group, mortality rates declined with an additional dose of HM in CIAV-infected chickens. HM supplementation in the diet of CIAV-infected chickens increased hematocrit (HCT), hemoglobin (Hb), and mean corpuscular volume (MCV) and decreased mean corpuscular hemoglobin concentration (MCHC) compared to the control (P < 0.05). Lymphocyte percentage and lymphocyte/heterophile ratio increased in HM-supplemented groups, especially at 2.5% (P < 0.05), and heterophile and granulocyte percentages were reduced (P < 0.05). Liver enzyme alkaline phosphatase (ALP) and liver steatosis declined in the 2.5% HM-treated group compared to the control (P < 0.05). It was concluded that adding 2.5% of the HM to the CIAV-infected broiler’s diet did not negatively affect chicken performance. In addition to its hypolipidemic effects, it could prevent HCT and Hb from decreasing in chicks infected with CIAV and positively affect leukocyte types and liver enzymes. Interestingly, an additional dose of HM in the diet of the CIAV-infected broilers reduced mortality. Therefore, adding 2.5% of HM could prevent the adverse effects of CIA on hematological traits in broiler chicken flocks without adverse effects on performance.

Virome Profiling of Chickens with Hepatomegaly Rupture Syndrome Reveals Coinfection of Multiple Viruses.

Liver diseases seriously challenge the health of chickens raised on scaled farms and cause tremendous economic losses to farm owners. The causative agents for liver diseases are still elusive, even though various pathogens, such as the hepatitis E virus, have been reported. In the winter of 2021, a liver disease was observed on a chicken farm in Dalian, China, which increased chicken mortality by up to 18%. We conducted panvirome profiling of the livers, spleens, kidneys, and recta of 20 diseased chickens. The viromic results revealed coinfection of multiple viruses, including pathogenic ones, in these organs. The viruses were highly identical to those detected in other provinces, and the vaccine and field strains of avian encephalomyelitis virus (AEV) and chicken infectious anemia virus (CIAV) cocirculated on the farm. In particular, the liver showed higher abundance of AEV and multiple fowl adenoviruses than other organs. Furthermore, the liver also contracted avian leukemia virus and CIAV. Experimental animals with infected liver samples developed minor to medium lesions of the liver and showed a virus abundance profile for AEV across internal organs similar to that in the original samples. These results suggest that coinfection with multiple pathogenic viruses influences the occurrence and development of infectious liver disease. The results also highlight that strong farm management standards with strict biosafety measures are needed to minimize the risk of pathogenic virus introduction to the farm.

Molecular characterization and pathogenicity study of a highly pathogenic strain of chicken anemia virus that emerged in China.

Chicken infectious anemia (CIA) is caused by chicken anemia virus (CAV). Recently, severe anemia has emerged in layer chickens (8 to 10-week-old) on poultry farms in China. However, the etiological characteristics and pathogenic potential of CAV in chickens at 6 weeks or older are not well understood. In this study, we isolated a CAV strain, termed SD15, from two-month-old chicken with severe anemia and analyzed the genetic evolution relationship. We found that strain SD15 had the highest homology (98.9%) with CAV18 strain. Comparison with 33 reference strains revealed 16 amino acid mutations in strain SD15, two of which were previously unknown (F210S in VP1 and L25S in Vp3). Compared with low pathogenic strains (Cux-1 and C14), highly pathogenic strains (SDLY08 and SD15) had three base mutations in their noncoding region. To further understand its pathogenicity, 10-week-old specific-pathogen-free (SPF) chickens were challenged with the novel strain and SDLY08. No obvious clinical symptoms were observed in the SDLY08 group. However, SD15-infected chickens showed significant growth retardation and immunosuppression. The main manifestations of immunosuppression were the significantly reduced thymus and bursa indices and AIV-H9 vaccine-induced antibody levels (P < 0.05). The lowest number of red blood cells in the SD15 group was just 60% of that in the control group. Taken together, the novel strain SD15 not only showed higher pathogenicity but also exhibited the potential ability to break the age resistance of older chickens to CAV. Our study enhanced the understanding of the epidemiological characteristics of chickens infected with severe anemia and can facilitate the development of improved control strategies of CIA in China.

Isolation, Identification, Sequence Analysis, and Pathogenicity of a CIAV Strain from Aegypius monachus

A cluster of vultures, artificially bred in a zoo in Shandong Province, China, displayed signs of emaciation and some even died, which raised the suspicion of an immunosuppressive pathogen. Upon conducting nucleic acid testing on the clinical samples, it was found that the CIAV was present in the tissue of dead vulture. In this study, samples of dead vulture tissue were used to isolate a chicken infectious anemia virus (CIAV) via the MDCC-MSB1 cell line, which was designated the SDTY2021-TJ strain. The full-length genomic sequence of SDTY2021-TJ was determined and analyzed in detail. The full genomic DNA of SDTY2021-TJ was found to be 2298 bp, with no omissions or additions in the coding region. The homology between the full-length genomic sequence of SDTY2021-TJ and the reference strain ranged from 95.5% (Del-Ros) to 98.7% (YN04). In comparison with the reference strain, the VP1 protein of SDTY2021-TJ contained a number of mutations. To assess the virulence of SDTY2021-TJ, one-day-old SPF chickens were inoculated with both high and low doses of the pathogen. The results demonstrated that SDTY2021-TJ had a considerable pathogenicity to SPF chickens, as the high-dose group caused a 50% mortality rate, and even the low-dose group caused a 30% mortality rate. The chickens infected with the disease exhibited paleness in the cockscomb and stunted growth, as well as a compromised response to NDV and AIV-H9 inactivated vaccine. Furthermore, histological observation revealed an atrophy and degeneration of the thymus. To the best of our knowledge, this is the first instance of CIAV being isolated from artificially bred Aegypius monachus, implying that wild birds may be involved in the transmission and spread of CIAV.

Retrospective Investigation and Genetic Variation Analysis of Chicken Infectious Anemia in Shandong Province, 2020–2022

Chicken infectious anemia (CIA) is a vertical transmission infectious chicken disease caused by the chicken infectious anemia virus (CAV). The disease can induce stunting and immunosuppression in chicks by infecting bone marrow-derived stem cells, causing huge economic losses for the poultry industry. To determine the prevalence of CIA in Shandong Province, China, 854 suspected CIA samples were collected and analyzed in 13 cities in Shandong from 2020 to 2022. The PCR results showed that a total of 115 CAV were isolated. The CAV-positive rates were 17.21% (26/151) in 2020, 12.23% (35/286) in 2021, and 12.94% (54/417) in 2022, with severe mixed infections. Among them, CAV and fowl adenovirus (FAdV) were the most common, accounting for 40.86%. VP1 gene homology analysis showed that isolated strains shared 96.1-100% homology with the previously reported CAV strains. Genetic variation analysis showed that most of the isolated CAV strains were located in genotype A. These results indicate that CIA infection in Shandong chickens in recent years has been prevalent and mixed infections are common, but there were no significant genetic variations. Our results extend the understanding of the prevalence and genetic evolution of CIA in Shandong Province. They will offer new references for further study of the epidemiology and virus variation and the prevention and control of this disease.

Genetic diversity, distribution, and evolution of chicken anemia virus: A comparative genomic and phylogenetic analysis.

Chicken infectious anemia (CIA) is an immunosuppressive poultry disease that causes aplastic anemia, immunosuppression, growth retardation and lymphoid tissue atrophy in young chickens and is responsible for huge economic losses to the poultry industry worldwide. The disease is caused by the chicken anemia virus (CAV), which belongs to the genus Gyrovirus, family Anelloviridae. Herein, we analyzed the full-length genomes of 243 available CAV strains isolated during 1991-2020 and classified them into two major clades, GI and GII, divided into three and four sub-clades, GI a-c, and GII a-d, respectively. Moreover, the phylogeographic analysis revealed that the CAVs spread from Japan to China, China to Egypt and subsequently to other countries, following multiple mutational steps. In addition, we identified eleven recombination events within the coding and non-coding regions of CAV genomes, where the strains isolated in China were the most active and involved in ten of these events. Furthermore, the amino acids variability analysis indicated that the variability coefficient exceeded the estimation limit of 1.00 in VP1, VP2, and VP3 proteins coding regions, demonstrating substantial amino acid drift with the rise of new strains. The current study offers robust insights into the phylogenetic, phylogeographic and genetic diversity characteristics of CAV genomes that may provide valuable data to map the evolutionary history and facilitate preventive measures of CAVs.

Chicken infectious anemia virus (CIAV) VP1 antagonizes type I interferon (IFN-I) production by inhibiting TBK1 phosphorylation

Chicken infectious anemia virus (CIAV) infection induces immunosuppression or subclinical immunosuppression in chickens. CIAV infection has been reported to repress type I interferon (IFN-I) expression, but the underlying mechanisms are not yet understood. Here we reported that VP1, the capsid protein of CIAV, the major immunogenic protein that triggers the production of neutralizing antibodies in chickens, inhibited type I interferon (IFN-I) expression induced by cGAS-STING signaling. We showed that VP1 inhibited TBK1 phosphorylation and down stream signal transduction, leading to the inhibition of IFN-I expression. Subsequently, we demonstrated that VP1 interacted with TBK1. Finally, we clarified that aa 120–150 in VP1 was essential for VP1 to interact with TBK1 and inhibit cGAS-STING signaling. These findings will help us further understand the pathogenesis of CIAV in chickens.

THE EFFECT OF NEEM LEAVES EXTRACT AS IMMUNSTIMULANT BEFORE CHICKEN INFECTIOUS ANEMIA VACCINATION IN BROILERS

Chicken infectious anemia (CIA) has lately arisen as a major problem in poultry production due to poor growth, high mortalities. We successfully used PCR on seven flocks of chickens (three breeder and four broiler) in the Egyptian governorates of Dakahlia and Damietta to detect CIAV in tissues of probable CIAV-infected birds. Numerous substances, including triterpenoids and glycosides, are found in Neem and are assumed to be the source of its antiviral effects. All birds were randomly allocated into eight groups (n=20 each) for different treatments, including vaccination, feeding Neem 8%, feeding infected birds 8%, vaccination and infected birds, feeding infected birds 8%, and finally feeding infected birds 8% and vaccination. One-hundred and sixty birds were randomly divided into eight groups (n=20 each). The groups were differentially-treated as the followings; healthy birds, birds infected with CIAV, birds fed Neem 8%, infected birds fed Neem 8%, vaccinated birds, vaccinated and infected birds, birds fed Neem 8% and vaccinated, and finally the infected ones fed Neem 8% and vaccinated. The study is aimed to detect the immunostimulant effect of Neem leaves extract 8% on immune response post vaccination against CIAV in broilers. The group taken immunstimulant (Neem leaves extract 8%) along with vaccination increased immune response of birds since titers of the enzyme-linked immunosorbent assay were 15200 (P≤ 0.05) than the group valued 14732 of vaccination only. However, in groups (Neem vaccinated infected, infected and vaccinated infected) were 14663, 12600 and 12091 (P&lt;0.05, P&lt;0.05 and P&lt;0.01, respectively). Infected group exhibited indications of CIAV as despondency, while group vaccinated and treated with Neem leaves extract 8% appeared normal after challenging with CIAV. The hematocrit values of infected group and vaccinated treated infected group were 21 and 30, respectively. Histopathological changes in Neem vaccinated group after challenging with CIAV showed increasing in the thickness of both cortex and medulla of thymic lobules beside over population. In the latest group, bone marrow showed activation and proliferated hemopoitic elements with regenerative centers. We conclude that the combination of Neem leaves extracts 8% and CIAV vaccination is a potent antiviral and has immunstimulant properties during the production cycle of broilers.