Primary growth plate chondrocyte isolation, culture, and characterization from the modern broiler.

Lameness is a leading cause of animal welfare and production concerns for the poultry industry as fast-growing, high-yielding broilers seem more susceptible to bone disease and infections. A major limitation to the study of these disorders is the lack of a chicken immortalized chondrocyte cell. Primary cell isolation is a valid and complex method for establishing a relevant in vitro model for diseases.

Thymosin β4 dynamics during chicken enteroid development.

The sheared avian intestinal villus-crypts exhibit high tendency to self-repair and develop enteroids in culture. Presuming that this transition process involves differential biomolecular changes, we employed matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) to find whether there were differences in the spectral profiles of sheared villi versus the enteroids, assessed in the mass range of 2-18 kDa. The results showed substantial differences in the intensities of the spectral peaks, one particularly corresponding to the mass of 4963 Da, which was significantly low in the sheared villus-crypts compared with the enteroids.

Sodium Butyrate Reduces Enteritidis Infection of Chicken Enterocytes and Expression of Inflammatory Host Genes .

Enteritidis (SE) is a facultative intracellular pathogen that colonizes the chicken gut leading to contamination of carcasses during processing. A reduction in intestinal colonization by SE could result in reduced carcass contamination thereby reducing the risk of illnesses in humans. Short chain fatty acids such as butyrate are microbial metabolites produced in the gut that exert various beneficial effects.

Production and characterization of avian crypt-villus enteroids and the effect of chemicals.

Background: Three-dimensional models of cell culture such as organoids and mini organs accord better advantage over regular cell culture because of their ability to simulate organ functions hence, used for disease modeling, metabolic research, and the development of therapeutics strategies. However, most advances in this area are limited to mammalian species with little progress in others such as poultry where it can be deployed to study problems of agricultural importance. In the course of enterocyte culture in chicken, we observed that intestinal mucosal villus-crypts self-repair and form spheroid-like structures which appear to be useful as ex vivo models to study enteric physiology and diseases.

Phorbol 12-Myristate 13-Acetate-Induced Changes in Chicken Enterocytes.

Increased intestinal epithelial permeability has been linked to many enteric diseases because it allows easy access of microbial pathogens and toxins into the system. In poultry production, the restrictions in the use of antibiotic growth promoters have increased the chances of birds being susceptible to different enteric diseases. Thus, understanding the mechanisms which compromise intestinal function is pertinent.

A method to culture chicken enterocytes and their characterization.

Enterocytes function as both absorptive and protective components of intestine that come in close contact with a variety of enteric factors, such as dietary, microbial, and parasites, that have potential to affect the organismal health. Understanding how enterocytes interact with this complex array of factors may help improve gut health particularly in the context of poultry production where it is also linked to food safety issues. The enterocyte in vitro culture can help screen different factors and their interactions with microbiome, and potentially be utilized in the development of interventions strategies for pathogens such as antibiotic alternatives.

Proteomic Changes in the Plasma of Broiler Chickens with Femoral Head Necrosis.

Femoral head necrosis (FHN) is a skeletal problem in broiler chickens, where the proximal femoral head cartilage shows susceptibility to separation from its growth plate. The selected birds with FHN showed higher body weights and reduced plasma cholesterol. The proteomic differences in the plasma of healthy and FHN-affected chickens were explored using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) and liquid chromatography/electrospray ionization-tandem mass spectrometry (LC-MS/MS) to prospect for protein biomarkers.

Proteomic Changes in Chicken Plasma Induced by Salmonella typhimurium Lipopolysaccharides.

Lipopolysaccharides (LPS) are cell wall components of Gram-negative bacteria that produce inflammation and sickness in higher animals. The objective was to identify plasma proteomic changes in an avian model of inflammation. Chickens were treated with either saline or LPS, and blood was collected at 24 hours postinjection.

Chicken Egg Shell Membrane Associated Proteins and Peptides.

Egg shells are poultry industry byproducts with potential for use in various biological and agricultural applications. We have been interested in the membranes underlying the calcareous shell as a feed supplement, which showed potential to improve immunity and performance of post hatch poultry. Therefore, to determine their protein and peptide profiles, we extracted the egg shell membranes (ESM) from fresh unfertilized eggs with methanol and guanidine hydrochloride (GdHCl) to obtain soluble proteins for analysis by mass spectrometry.

Effect of thiram on avian growth plate chondrocytes in culture.

Thiram is a dithiocarbamate pesticide that causes tibial dyschondroplasia (TD), a growth plate defect, in poultry. Deaths of transitional zone chondrocytes appear to interrupt endochondral bone development leading to the broadening of growth plate. The mechanism of action of thiram on chondrocytes is not well understood.

Effect of toll-like receptor activation on thymosin beta-4 production by chicken macrophages.

Thymosin beta-4 (Tβ4) is an actin-binding intracellular peptide that promotes wound healing, tissue remodeling, and angiogenesis. The mechanism of Tβ4 secretion to the extracellular environment is not understood. The macrophage is a rich source of Tβ4 which also participates in wound healing process.

Evaluation of beta defensin 2 production by chicken heterophils using direct MALDI mass spectrometry.

Beta defensins (BD) are cysteine rich, cationic antimicrobial peptides (AMP) produced mainly by epithelial and myeloid cells such as neutrophils. In birds, the neutrophil equivalent heterophils produce avian beta defensins (AvBD) of which AvBD2 is the major isoform. Heterophils recognize pathogens or their derived products through a series of pattern recognition receptors called toll-like receptors (TLR) leading to their antimicrobial activities.

Identification and characterization of thymosin beta-4 in chicken macrophages using whole cell MALDI-TOF.

The aim of the study was to determine chicken monocyte- and granulocyte-associated peptides and proteins using "whole cell" matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and to characterize the peptides based on their abundance. The mass spectra showed a prominent peak at m/z 4963 in monocytes/macrophages but not in the granulocytes. Subsequent purification and characterization of the m/z 4963 peptide from an avian macrophage cell line HTC, revealed it to be thymosin beta-4 (Tbeta-4), an actin-modulating peptide.

Effects of ovotransferrin on chicken macrophages and heterophil-granulocytes.

Ovotransferrin (OTF) is an acute phase protein in chickens, serum levels of which increase in inflammation and infections. To understand the significance of OTF in inflammation, we studied its in vitro effects on HD11 cells, a macrophage cell line, and heterophils isolated from blood using a panel of variables indicative of cellular activation. These included the production of interleukin-6 (IL-6), nitrite, matrix metalloproteinase (MMP), oxidation of dichlorofluorescein diacetate for respiratory burst and the degranulation of heterophils by the loss of fluorescein isothiocyanate positive cytoplasmic granules.