Although intensive vaccination programs have been implemented, Newcastle disease (ND) outbreaks, accompanied by severe economic losses, are still reported in Egypt. The genetic characterization of ND virus (NDV) strains isolated from ND-vaccinated chicken flocks provides essential information for improving ND control strategies. Therefore, here, 38 NDV strains were isolated and identified from outbreaks among vaccinated flocks of broiler chickens located in the provinces of Qena, Luxor, and Aswan of Upper Egypt during 2011-2013. The investigated broiler chicken flocks (aged 28 to 40 days) had high mortality rates of up to 80%. All NDV isolates were genetically analyzed using next-generation DNA sequencing. From these isolates, 10 representative NDV strains were selected for further genetic analyses. Phylogenetic analysis of full-length coding genes revealed that the Egyptian NDV isolates belonged to a single sub-genotype, VII.1.1. These isolates were phylogenetically distant from the vaccine strains, including La Sota or Clone 30 (genotype II), which have been commonly used to vaccinate chicken flocks. Amino acid substitution K78R was observed in the neutralizing epitopes of the F proteins; whereas several mutations were found in the neutralizing epitopes of the hemagglutinin-neuraminidase proteins, notably, E347K. Overall, our results suggested that the occurrence of neutralizing epitope variants may be one of potential reasons for ND outbreaks. Further studies are needed to determine the protective effect of current vaccines against circulating virulent NDV strains.

Vaccination against Newcastle disease (ND), a devastating viral disease of chickens, is often hampered by thermal inactivation of the live vaccines, in particular in tropical and hot climate conditions. In the past, “thermostable” vaccine strains (I-2) were proposed to overcome this problem but previous comparative studies did not include formulation-specific factors of commercial vaccines. In the current study, we aimed to verify the superior thermal stability of commercially formulated I-2 strains by comparing six commercially available ND vaccines. Subjected to 37 °C as lyophilized preparations, two vaccines containing I-2 strains were more sensitive to inactivation than a third I-2 vaccine or compared to three other vaccines based on different ND strains. However, reconstitution strains proved to have a comparable tenacity. Interestingly, all vaccines still retained a sufficient virus dose for protection (106 EID50) after 1 day at 37 °C. These results suggest that there are specific factors that influence thermal stability beyond the strain-specific characteristics. Exposing ND vaccines to elevated temperatures of 51 and 61 °C demonstrated that inactivation of all dissolved vaccines including I-2 vaccine strains occurred within 2 to 4 h. The results revealed important differences among the vaccines and emphasize the importance of the quality of a certain vaccine preparation rather than the strain it contains. These data highlight that regardless of the ND strain used for vaccine preparation, the appropriate cold chain is mandatory for keeping live ND vaccines efficiency in hot climates.

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Objective: The purpose of the current study was to evaluate the molecular characteristics of naturally mutant non-pathogenic O27 strain of Escherichia coli and its efficacy as probiotic in broilers and determine the best age at which it can be administered. Materials and methods: A total of 24 virulence genes using 24 sets of primers were detected using the polymerase chain reaction technique. For probiotics experiments, 60 chicks (day 1 old) were divided into three groups, 20 per group, and reared for 4 weeks. The first group was considered as a negative control. The second group was treated orally with O27 strain at first day of life for three successive days and repeated at day 21. The third group was administered orally with O27 strain at day 10 old, and repeated at day 21 old. Results: The data revealed that type 1 fimbrial adhesion, salmochelin siderophore receptor, and sigma factor-binding protein were detected in O27 strain, but temperature-sensitive hemagglutinin, hemolysin secretion gene, pyelonephritis-associated Pili gene, polysaccharide capsule synthesis gene, Shiga-toxin1 gene, Shiga-toxin2 gene, Brain microvascular endothelial cell invasion, E. coli attaching and effacing gene, heat-stable enterotoxin, heat-labile enterotoxin, east 1 toxin, colicin V, verotoxin type 2, necrotizing cytotoxic factor type 1, colonization factor antigen I, colonization factor antigen III, coli surface 2, coli surface 4, serine protease pic autransporter, vacuolating autotransporter toxin, and serine protease EspP precursor were not detected in O27 strain. Group 2 performance parameters were significantly better ( p < 0.01 ) than groups 3 and 1. Hematological and biochemical parameters did not be influenced ( p > 0.05 ) by the administration of O27 strain. Antibody titers of infectious bursal disease virus and Newcastle disease virus in groups 2 and 3 were improved as compared to group 1. Group 2 had significantly higher titers than group 3. Histopathologically, all groups showed normal histopathological pictures. However, jejunum in groups 2 and 3 showed more tall, intact, and densely packed microvilli and more crypt depth than the control group. Conclusion: The O27 strain of E. coli is non-pathogenic bacteria. Its effects on growth performances and enhancement of immunity in broilers match with the same impact of probiotics, and these candidates will fit to be a good probiotic in the future. The results revealed that the effects of O27 strain at the day 1 old of life for three successive days and repeated at day 21 old are better for improving the performance and immunity of the birds. More research works about the characterized non-pathogenic E. coli strain O27 are required for field and commercial use.

The infectious bursal disease (IBD) is a continuing serious problem facing poultry industry in Egypt. In this study, 500 bursae samples were collected from different broiler flocks in different localities of Luxor Governorate. The flocks were suffering from mortality and bursal lesions during Dec.2014 to Jan. 2016. The samples were tested by AGPT and the result was 12 flocks only were positive for IBD and then the positive samples were isolated in SPF embryonated chicken eggs. All the inoculated embryos died within 2-3 days. The embryos were smaller than normal, congested with haemorrhagic head, and necrotic foci of liver. Then the presence of virus in embryonated eggs was confirmed by convential PCR. Furthermore, molecular characterization was performed by direct sequencing of a 620-bp cDNA corresponding to the VP2 variable domain of the polyprotein gene synthesized by PCR. With deduced amino acid analysis found that all examined isolates are very virulent strains. four local strains used for nucleotide sequence, percent identity and phylogenetic tree analysis revealed that four isolates (F21,F23,F24,F26) were very close to very virulent old Egyptian strains Giza 2008.

A total of 42 Newcastle disease virus (NDV) strains were isolated from clinical samples obtained from chickens during field outbreaks among vaccinated broiler chicken's farms in the Upper Egypt in 2011 -2012. The samples were inoculated into embrayonated chicken eggs and the positive samples for NDV were detected by hemagglutinination (HA) and confirmed by hemagglutination inhibition (HI) tests and reverse transcriptation polymerase chain reaction (RT-PCR). The pathogenicity of the isolates was estimated biologically by intracerebral pathogenicity index (ICPI) test and genetically by sequencing the partial fusion (F) genes. The ICPI indicated that 40 isolates were > 1.5, which characterized velogenic strains, the remaining was mesogenic (ICPI = 1.4). The sequencing analysis of the nucleotides and deduced amino acids, 8 isolates representing the isolates from different localities, revealed that these isolates possess the amino acids motif 112-R-R-Q-K-R-F-117 of the velogenic nature and a diversity of 19 -22 and 19-21 % in nucleotides and amino acids, respectively, from vaccine strains (LaSota, B1, Ulster/67 and I-2) and Texas/GB. Interesting, that these isolates clustered into two subgroups related to the isolates from Jordan, Israel (2011) and China but were distant from vaccine, Egyptian (2005, 2006 and 2010), Sudan, Saudi Arabia and Israel (2003 strains by phylogenetic analysis. In conclusion, a velogenic NDV prove to circulate among chickens in southern of Egypt although intensive vaccination programs were conducted. These isolates were genetically away from the vaccine strains and virulent to chickens. The necessity needs to continuously monitor the virus and evaluate the vaccination programs efficiency.

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