Alveolar macrophages (AMs) respond to these infections as the first line of defense and trigger the lung's immune response. Knowing the expression dynamics of key immune-related genes in alveolar macrophages against M. haemolytica challenge will help deepen the understanding of disease immunopathogenesis. For this purpose, first time an in vitro obtained from bronchoalveolar lavage (BAL) fluid and treated with three doses (T1: 1800, T2: 2700, T3: 5400 CFU) of M. haemolytica inoculum, while keeping one untreated as a control. Then, total RNA was extracted, and cDNA was synthesized. The real-time quantification data indicated that the expression of IL-1β, IL-6, IL-8, IL-10, and NF-κB genes were significantly changed as compared to control. Our study revealed that the exposure to M. haemolytica stimulates the immune response in the sheep alveolar macrophages in a dose and time-dependent manner.

Background/Objectives: Hatchery fumigation is recognized as a crucial step to control microbial bloom in the environment, and formaldehyde is one of the most widely used disinfectants to ensure successful hatchability and healthy production. While many of the benefits are thought to be derived from disinfectant properties, it is possible that additional host gene and genetic pathway modulation could contribute to these outcomes. The current study aimed to capture the in ovo transcriptional response of liver tissue to formaldehyde treatment. Methods: Chick embryos were subjected to formaldehyde fumigation treatment for 25 min at 24–25 °C and 75% relative humidity, keeping a control group as untreated. On the 18th day of incubation at 37.8 °C and 58–63% humidity, eggs were broken, and liver tissue was obtained for RNA isolation, cDNA library preparation, and RNA sequencing. Results: Bioinformatics analysis revealed 908 significant differentially expressed genes (DEGs), among which 814 were known genes and 94 were novel genes. A total of 672 DEGs were upregulated, whereas 236 genes were downregulated in response to FA treatment. Of the 94 novel genes, 80 were upregulated. Key DEGs, associated QTLs, and transcription factors were involved in immuno-inflammatory responses, oxidative stress, epigenetic modification, and cellular adaptation-related activities. Further research should focus on biological validation of key DEGs to clarify their roles, pathways, and relationships to FA treatment. Conclusions: Overall, these findings (1) provide critical molecular detail as a first step towards genetic selection to improve formaldehyde treatment response and effectiveness, and (2) provide DEG signatures for FA treatment as a reference against which to compare other interventions to achieve hatchability and production benefits.

Formaldehyde (FA) is commonly used for hatchery disinfection, where it reduces microbial growth, ensures successful egg hatch and enhances healthy production, but its specific effects on embryonic development remain unclear. MicroRNAs (miRNAs) regulate gene expression post-transcriptionally and may mediate FA-induced transcriptional responses. Here, we investigated the impact of FA treatment on miRNA profiles in chicken embryo liver. Small RNA-seq libraries were constructed and sequenced using the Illumina NextSeq platform. Reads were trimmed and quantified using miRDeep2 version 2.0.0.3. Differential expression analysis was performed with DESeq2 (p-adjusted < 0.05 and |log₂FC| > 1). Target genes of differentially expressed miRNAs (DEMs) were predicted with miRDB, and GO/KEGG/Reactome enrichment was conducted. Out of 662 total mature miRNAs detected, differential expression analysis identified 30 DEMs (11 up-regulated, 19 down-regulated). The highest fold increase was determined for gga-miR-3533 (log₂FC = 4.45), and the most significant decrease was determined for gga-miR-133b (log₂FC = −3.38). Pathway analysis revealed miRNAs affecting signaling pathways along with modules related to post-translational protein modification, immune system, and oxidative stress pathways. Our study demonstrates that FA treatment can affect critical biological processes by altering miRNA-mediated regulation in the developing embryonic liver and point to the need for functional validation of miRNA-target interactions to help determine mechanisms for FA benefits. Long term, these data may help serve as reference to identify new treatments with optimized response profiles.

The global human population is projected to reach 9.7 billion by 2050, placing an unprecedented burden on food systems and amplifying the need for sustainable poultry production. Free-range and organic poultry farming are gaining popularity due to consumer demand for animal welfare and environmental sustainability. However, these systems present unique challenges with regard to therapeutic strategies, bioactive environmental exposure and their degradation. Long-acting therapeutic strategies are needed to minimize chicken handling and stress, while also ensuring therapeutic efficiency in open environments. Controlled-release delivery systems (CRDS) offer significant potential to address these challenges by prolonging therapeutic activity, protecting bioactive from environmental degradation, and reducing labor and animal stress. CRDS can enable efficient delivery of drugs, vaccines, botanicals and probiotics. Despite these advantages, implementing controlled-release technologies in free-range and organic settings is a complex process and presents many challenges. We investigated the current state of CRDS for small molecules, vaccines, plant botanicals, and probiotics, highlighting key challenges and future prospects for their application in the evolving poultry industry.

The alveolar macrophages (AMs) are frontier of defense against foreign materials that initiate immune response in lungs. Knowledge of the expression dynamics of major immune-related genes in the alveolar macrophages in response to lipopolysaccharide (LPS) and lipoteichoic acid (LTA) challenge can help to understand disease mechanism involved in several respiratory diseases. The aim of this study was to investigate the mRNA expression of selected immune-related genes in response to lipopolysaccharide (LPS) and lipoteichoic acid (LTA) challenge in sheep alveolar macrophages in vivo. Results revealed that Romanov lambs exhibited higher mRNA expression of TLR2, TLR4, NF-ĸβ, TNFα, IL-1β, IL-6, IL-8, and IL-10 genes as compared to Akkaraman lambs along with the control of all treatments. Moreover, the expression of TLR2, TLR4, NF-ĸβ, TNFα, IL-1β, IL-6, IL-8, and IL-10 genes was higher in combine treatment of LPS and LTA as compared to separate treatments of LPS and LTA in both breeds. The results showed that the mRNA expression of immune-related genes was significantly increased in the sheep AMs in response to LPS and LTA treatment whereas a synergistic effect was observed in LPS+LTS treatment. Also, breed comparison showed that the native Akkaraman was more resistant to disease compared to exotic Romanov.