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Traditional approaches to assess the immune response of chickens to infection are through animal trials, which are expensive, require enhanced biosecurity, compromise welfare, and are frequently influenced by confounding variables. Since the chicken embryo becomes immunocompetent prior to hatch, we here characterized the transcriptional response of selected innate immune genes to Newcastle disease virus (NDV) infection in chicken embryos at days 10, 14, and 18 of embryonic development. The results suggest that the innate immune response 72 h after challenge of 18-day chicken embryo is both consistent and robust. The expression of CCL5, Mx1, and TLR3 in lung tissues of NDV challenged chicken embryos from the outbred Kuroiler and Tanzanian local ecotype lines showed that their expression was several orders of magnitude higher in the Kuroiler than in the local ecotypes. Next, the expression patterns of three additional innate-immunity related genes, IL-8, IRF-1, and STAT1, were examined in the highly congenic Fayoumi (M5.1 and M15.2) and Leghorn (Ghs6 and Ghs13) sublines that differ only at the microchromosome bearing the major histocompatibility locus. The results show that the Ghs13 Leghorn subline had a consistently higher expression of all genes except IL-8 and expression seemed to be subline-dependent rather than breed-dependent, suggesting that the innate immune response of chicken embryos to NDV infection may be genetically controlled by the MHC-locus. Taken together, the results suggest that the chicken embryo may represent a promising model to studying the patterns and sources of variation of the avian innate immune response to infection with NDV and related pathogens.

Newcastle disease is a devastating viral disease of chicken in low- and middle-income countries where the backyard production system is predominant. Marker-assisted selection of chickens that are resistant to Newcastle disease virus (NDV) is the promising strategy that needs to be explored. The aim of the present study was to investigate polymorphisms of the promoter region of the chicken Mx gene and association with Kuroiler, Sasso, and local Tanzanian chicken embryos' survival variability to virulent NDV infection. Chicken embryos were initially challenged with a minimum lethal dose of virulent NDV suspension and then were followed over time to gather information on their survival variability. Using the survival data, high and less susceptible cohorts were established, and a total of 88 DNA samples from high and less susceptible groups were genotypes by sequencing. Five single-nucleotide polymorphisms (SNPs), which were previously reported, were detected. Interestingly, for the first time, the findings demonstrated the association of the promoter region of chicken myxovirus-resistance (Mx) gene polymorphisms with chicken embryos' susceptibility to the virulent NDV challenge. At the genotypic level, the SNP4 G > A mutation that was located within the IFN-stimulating response element was associated (LR: 6.97, P=0.03) with chicken embryos' susceptibility to the virulent NDV challenge. An allele G frequency was higher in the less susceptible cohort, whereas an allele A frequency was higher in the high susceptible cohort. At the haplotype level, the haplotype group ACGC was associated (OR: 9.8, 95% CI: 1.06–79.43, P=0.042) with the same trait and had a resistant effect. In conclusion, the results have demonstrated the association of chicken Mx gene promoter polymorphisms and chicken embryos' survival variability to the virulent NDV challenge, and the information is useful for breeding programs designed to develop chicken genotypes that are resistant to Newcastle disease virus.

Medicinal plants have been the good source of treatment for different ailments of humans as well as animals for centuries. However, in Tanzania, few plants were investigated for their efficacy against various diseases of chickens. In the present study, four medicinal plants were investigated against Salmonella gallinarum isolated from chickens. The minimum inhibitory concentration (MIC) using the broth microdilution methods and minimum bactericidal concentration (MBCs) were used to evaluate the activities of plants against chicken salmonellosis. For the safety of chickens against the toxicity of plants, the cytotoxicity assay was determined using a brine shrimp lethality test. Aloe secundiflora leaf ethyl acetate (ALEA), Aloe rabaiensis leaf methanolic (ArM), Aloe rabaiensis leaf ethyl acetate (ArLEA), and Punica granatum leaf ethyl acetate (PGLEA) extracts exhibited the highest MIC (0.3906 mg/mL) and MBC (3.125 mg/mL), respectively. The Dolichos kilimandscharicus tuber ethyl acetate (DTEA) and Dolichos kilimandscharicus tuber pet ether (DTPE) extracts displayed MIC of 1.563 mg/mL and 12.50 mg/mL and MBC of 12.50 mg/mL and 25.50 mg/mL, respectively. The highest LC50 values exhibited in Dolichos kilimandscharicus ranged from 7.937 × 10−4 mg/mL to 7.242 × 10−2 mg/mL for pet ether and methanolic extracts, respectively, while ALEA extract exhibited LC50 of 7.645 × 10−3 mg/mL. Generally, the extracts with MIC 0.3906 mg/mL and MBC 3.125 mg/mL demonstrated the highest antibacterial activity with low toxicity efficient to manage chicken salmonellosis. Dolichos kilimandscharicus, which exhibited higher toxicity, warrants further investigation on insecticidal and anticancer agents.

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