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The continued circulation of infectious bursal disease virus (IBDV) in Egypt, despite the use of various vaccines, is a serious problem that requires continuous detection of IBDV. In the current study, real-time reverse transcriptase polymerase chain reaction testing of 100 diseased chicken flocks during 2017-2021 revealed the presence of very virulent IBDV (vvIBDV) in 67% of the flocks, non-vvIBDV in 11%, and a mixture of both vvIBDV and non-vvIBDV in 4%. Twenty-nine IBDV isolates were submitted for partial sequencing of the viral protein 2 hypervariable region (VP2-HVR), and 27 isolates were confirmed to be genogroup A3 (vvIBDV) with 96.3%-98.5% similarity to the global A3 (vvIBDV) and 88.9%-97% similarity to genogroup A1 vaccine strains. The remaining two isolates were non-vvIBDV and showed 91.1% and 100% identity with classical genogroup A1 strains, respectively. Furthermore, the sequence and phylogenetic analysis of VP1 (amino acids 33-254) of two selected isolates of A3, 5/2017 and 98/2021, clustered them as B2, vvIBDV-like, strains with high similarity (99.5%) to four Egyptian, 99% to Chinese and European, and 97.7% to Chinese and Polish vvIBDV isolates. Experimental infection of commercial broiler chickens with two vvIBDV-A3B2 isolates (5/2017 and 98/2021) showed no mortality despite typical tissue lesions, clear histopathological changes, and strong ELISA antibody response. Isolate 98/2021 was more pathogenic, as confirmed by histopathology, whereas isolate 5/2017 induced a stronger serological response. In conclusion, vvIBDV (A3B2) strains with two amino acid (aa) substitutions in VP1 as V141I and V234I as well as VP2 as Y220F and G254S are still circulating in Egypt.

Despite the vast Egyptian poultry production, scanty information is available concerning the infection of haemprotozoan parasites as pathogens in commercial broilers. In the present study, we provided the first detection of leucocytozoonosis in five broiler chicken flocks in El-Beheira Egyptian governorate. Despite the low mortality rates in the affected flocks (0.3%-1% as a 5-day mortality), severe postmortem (hemorrhagic spots and scars) and histopathologic lesions appeared in different organs including skeletal muscles, liver, kidney, pancreas, abdominal cavity, and bursa of Fabricius. Evaluation of blood smears revealed gametocytes in erythrocytes and leukocytes. Conventional reverse transcriptase-PCR and partial sequence analysis of mitochondrial cytochrome oxidase b gene detected Leucocytozoon caulleryi . GenBank accession numbers of the five Egyptian L. caulleryi isolates were obtained. The five L. caulleryi were 99.9% identical to each other and 99.14% similar to the L. caulleryi mitochondrial DNA gene of Asian strains from India, Japan, Malaysia, South Korea, Taiwan, and Thailand.

Newcastle disease virus (NDV), a major pathogen of poultry worldwide, causes significant economic losses in the poultry industry. To characterize the ability of recently isolated virulent strains of NDV genotypes VI and VII to cause disease in quails, and to evaluate the efficacy of two NDV vaccines against such strains, Japanese quails were experimentally inoculated with either NDV genotype VI (Pigeon F-VI strain) or VII 1.1 (GHB-328 strain) with or without vaccination with inactivated NDV vaccine of genotype II (La Sota strain) or VII (KBNP strain). Mild to severe neurological signs developed in quails inoculated with the Pigeon F-VI strain from 3 to 14 days post infection (PI) and from 4 to 10 days PI in birds infected with the GHB-328 strain. The mortality rates were 46% and 33% for birds inoculated with NDV VI and NDV VII 1.1, respectively. The severity of histopathological changes depended on the viral isolates used. Vaccination with the La Sota or KBNP vaccine strain successfully protected quails against NDV-induced mortality and decreased the severity of clinical signs, pathological changes and cloacal viral shedding. This study showed that these virulent NDV isolates had mild to moderate pathogenicity in quails and that both vaccines protected against challenge with both virus strains. NDV vaccine genotype VII improved the level of protection against challenge with the VII 1.1 genotype compared with the classic vaccine, but failed to protect quails against challenge with the VI genotype., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The prevalence of Gallibacterium anatis in poultry production has increased over the last two decades. However, only a few studies have explored the pathogenicity of this bacterium in commercial layer chickens. This trial studied the aspects of the pathogenicity of a Gallibacterium anatis biovar haemolytica local Egyptian isolate (previously registered as strain B14 with GenBank accession no. KJ026147). We used 500 base pairs of a 16S ribosomal RNA gene and the 16S-23S ribosomal RNA intergenic spacer, partial sequence in an experimental infection trial in commercial White Shaver layer chickens aged 19 wk. The hens were divided into three groups of 40 birds each. The hens in Groups 1 and 2 were experimentally infected through the intranasal (IN) and intravenous (IV) routes, respectively, with a dose of 0.2 ml/bird containing 1.2 × 109 colony-forming units/ml. In contrast, Group 3 was kept as a noninfected control group. Both IN and IV infections resulted in a delayed egg laying for 1 wk and a significant (P ≤ 0.05) drop in egg production by 7.81% and 10.28% compared with the control group over 7 wk. Severe lesions in the form of hemorrhagic pneumonia, catarrhal tracheitis, ovarian follicle and oviductal regression, and septicemia were evident on necropsy, demonstrating the pathogenicity of G. anatis as a primary pathogen.

Background: Newcastle disease (ND) causes severe economic losses in poultry industry worldwide. Egyptian poultry industry suffered from severe economic losses since the isolation of Velogenic Newcastle disease virus (vNDV) genotype VIId in 2011 and up till now despite the use of different vaccination programs. So, this study aimed to isolate and characterize the vNDV from a total of 120 poultry flocks from ten provinces in the Egyptian Delta region with a history of respiratory manifestation, high mortalities or a decrease in egg production between 2015 and 2019. Seventy-three samples' allantoic fluid (73/120, 60.8%) were positive for hemagglutination with chicken RBCs. These samples were submitted to molecular examination using qRT-PCR specific primers for AOAV-1, highly pathogenic avian influenza (HPAI-H5), low pathogenic avian influenza (LPAI-H9) and infectious bronchitis virus (IBV)., Results: Fifty samples (50/120: 41.6%) were confirmed positive for AOAV-1, based on genetic analysis of matrix and fusion protein. The co-infection rate of other respiratory viral diseases examined was 1.6, 14.1, and 4.1%, for HPAI-H5, LPAI-H9, and IBV, respectively. Biologically, the intracerebral pathogenicity index of ten selected AOAV-1 isolates ranged from 1.70 to 1.98, which indicated the velogenic nature of these isolates. All the sixteen sequenced isolates were AOAV-1 genotype VII.1.1. The full F gene sequence of six examined AOAV-1 VII.1.1 isolates contained the seven neutralizing epitopes, and the glycosylation motif of six-potential sites for N linked glycosylation at residues 85, 191, 366, 447, 471, and 541., Conclusion: It could be concluded that the high prevalence of AOAV-1 genotype VII.1.1 in the Egyptian chicken flocks despite the intensive vaccination with live and killed ND vaccines, as all the 16 isolates tested were belonged to this genotype. Homologous vaccination is badly needed to control and reduce the spread of AOAV-1 genotype VII.1.1infection in Egyptian poultry flocks.

This work was designed to study the dynamics of transmission of Newcastle disease virus (NDV), genotype VIId, from Muscovy ducks (Cariana moscata) infected either by intramuscular (IM) or intranasal (IN) inoculation, to in-contact broiler chickens (Gallus gallus). IM-infected Muscovy ducks (G1d) exhibited only 5% mortality, and the concentration of virus shed from the cloaca was greater and for longer period than virus shed from the trachea. In contrast, IN-infected ducks (G2d) exhibited no mortality, and virus shedding from the trachea was higher than that from the cloaca starting from 4 days post infection (dpi) and continued up to 16 dpi, while in IM-infected ducks (G1d), tracheal shedding stopped at 11 dpi. Chickens in contact with IM-infected and IN-infected ducks, G1c and G2c, respectively, not only developed severe clinical symptoms and death (80% and 20% mortality, respectively), but also shed the virus at higher concentrations than infected ducks. G1c chickens had higher viral shedding titers in both the trachea and cloaca than G2c chickens until 11 dpi. All broiler chickens infected by IM route (G3c) died, while the IN route of infection resulted in lower mortality (70%) in G4c. Generally, all IM-infected birds produced an earlier and higher level of NDV hemagglutination inhibition (HI) antibody titer, along with higher rates and shorter periods of viral shedding than those infected by the intranasal route. Our conclusion is that Muscovy ducks are efficient carriers of NDV-genotype VIId and transmit the virus to contact chickens., (© 2019 Poultry Science Association Inc.)

This study was conducted to perform the comparative molecular characterization of avian influenza virus (AIV) H9N2, pathogenicity and seroprevalence in commercial and backyard poultry flocks. Fifty commercial poultry flocks were investigated between 2012 and 2015. Eighteen flocks (36%) out of 50 were positive HA. Seven (38.9%) out of 18 were positive by chromatographic strip test for AI common antigen. By Real-time RT-PCR, only two flocks were positive H9. The molecular characterization of two different AI-H9N2 viruses, one isolated from a broiler flock (A/chicken/Egypt/Mansoura-18/2013) and the other from a layer flock (A/chicken/Egypt/Mansoura-36/2015) was conducted on HA gene. Moreover, a higher seroprevalence, using the broiler strain as a known antigen, was shown in backyard chicken flocks 15/26 (57.7%) than duck flocks 9/74 (12.2%). Interestingly, the pathogenicity index (PI) of the H9N2 broiler strain in inoculated experimental chickens ranged from 1.2 (oculonasal route) to 1.9 (Intravenous route). The PI indicated a highly pathogenic effect, with high mortality (up to 100%) in the inoculated chickens correlated with the high mortality (80%) in the flock where the virus was isolated. The firstly recorded clinical signs, including cyanosis in the combs and wattles and subcutaneous haemorrhages in the leg shanks and lesions, as well as histopathology and immunohistochemistry, revealed a systemic infection of the high pathogenicity with the H9N2 virus. Conversely, the H9N2 layer strain showed a low pathogenicity. In conclusion, as a first report, the molecular analysis and pathogenicity of the tested strains confirmed the presence of a high pathogenicity AIV-H9N2 with systemic infections., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: The Avian Influenza A (H5N1) virus is endemic in poultry in Egypt. The winter of 2014/2015 was particularly worrying as new clusters of HPAI A (H5N1) virus emerged, leading to an important number of AI A (H5N1) outbreaks in poultry farms and sporadic human cases. To date, few studies have investigated the distribution of HPAI A (H5N1) outbreaks in Egypt in relation to protective / risk factors at the farm level, a gap we intend to fill. The aim of the study was to analyse passive surveillance data that were based on observation of sudden and high mortality of poultry or drop in duck or chicken egg production, as a basis to better understand and discuss the risk of HPAI A (H5N1) presence at the farm level in large parts of the Nile Delta., Results: The probability of HPAI A (H5N1) presence was associated with several characteristics of the farms. Vaccination status, absence of windows/openings in the farm and the number of birds per cycle of production were found to be protective factors, whereas the presence of a duck farm with significant mortality or drop in egg production in the village was found to be a risk factor., Conclusions: Results demonstrate the key role of several prevention and biosecurity measures to reduce HPAI A (H5N1) virus circulation, which could promote better poultry farm biosecurity in Egypt.

Gallibacterium anatis biovar haemolytica constitutes a part of the normal microflora in the upper respiratory and genital tracts of healthy chickens, but it is also associated with different pathological conditions. In the current study, 102 commercial chicken flocks suffering from respiratory disease and/or drop in egg production were investigated for the presence of G. anatis during 2013 and 2015. These flocks comprised 8 breeder, 32 layer, and 62 broiler flocks. By culture method, 20 flocks were found positive: one isolate derived from broiler breeders, 6 isolates from layers, and 13 isolates from broilers. G. anatis biovar haemolytica was identified by phenotyping and PCR. Additionally, partial genome sequencing of 11 isolates (5 layer isolates of 2013 and 6 broiler isolates of 2015) based on 16S rRNA and 23S rRNA gene sequences was performed and revealed 96.5% to 100% genetic relatedness. Antibiotic sensitivity of these isolates revealed that the 2013 isolates were highly susceptible to florfenicol while the isolates of 2015 were highly susceptible to cefotaxime. Gallibacterium anatis biovar haemolytica is a newly introduced bacteria in Egypt causing salpingitis, peritonitis, drop in egg production, and/or respiratory signs.

Marek's disease is a lymphoproliferative disease causing a serious threat in poultry production. Field strains of Marek's disease virus (MDVs) are continuously re-emerging, causing great economical losses to the poultry industry worldwide in spite of the intensive vaccination and restrictive management policy used. Histopathological and molecular characterizations of MDVs are essential for monitoring the changes of viruses and evaluating the effectiveness of existing vaccines. During 2016, 190 visceral tumour tissues representing 30 vaccinated chicken flocks from the Gifu prefecture, Japan, were analysed. A pathological examination revealed the presence of lymphoproliferative lesions in the visceral organs. Polymerase chain reaction screening of tissue specimens using specific primers for avian leucosis virus, reticuloendotheliosis virus, and MDV was positive only for MDV. The polymerase chain reaction products of meq, pp38, virus-induced IL-8 homology, and glycoprotein MDV genes were sequenced and used for homology, phylogenetic, and similarity level analysis with the published reference of MDVs in the database. The results revealed high similarity between the field isolates, vv and vv+ strains of MDV from the USA and China. Several point mutations in the nucleotide sequence of the field isolates and their deduced amino acid sequences were detected in those genes. The present molecular analyses indicated that nucleotide and amino acid changes could be valuable criteria for differentiation and determination of the pathogenicity and oncogenicity of MDVs according to the Avian Disease and Oncology Laboratory pathotyping in vivo studies. Furthermore, the results suggest that development of a new vaccine must be considered to overcome this devastating avian oncogenic viral disease.

The present study was conducted to evaluate the adverse effects of an interaction between low levels of dietary aflatoxins (AF) and Eimeria tenella infection on broiler chicks. A set of 1-day-old chicks were raised for 35 days in the following groups: a control group, a group fed AF, a group fed AF and inoculated with E. tenella (AF + E.ten), and a group inoculated with E. tenella alone. AF in the contaminated diet were given at 200 ppb starting from the seventh day after hatching while E. tenella was inoculated at a dose of 5 × 10(4) sporulated oocysts per chick at the 14th day after hatching. Worsened performance traits and high mortality were all observed in the treated birds, particularly the AF + E.ten group. Lesion scores and oocyst outputs were not different within groups. Chickens fed with AF had significantly increased serum ALT and ALP activities as well as decreased albumin content. They also showed hepatomegaly, hepatocytic vacuolation and necrosis, an atrophied bursa of Fabricius, and a thymus with tissue depletion. E. tenella-infected broilers displayed a significant reduction in packed cell volume, hemoglobin content and lymphocyte percentage, and showed hemorrhagic typhlitis. The deficits in hepatic function and hematologic parameters as well as the gross pathological, and histopathological changes, were more common and more severe in the group that was exposed to both aflatoxicosis and coccidiosis than in the groups exposed to either treatment alone. Thus, the combination of aflatoxicosis and E. tenella infection may influence the course of coccidial infection due to additive effects.