The objective of this study was to assess the molecular impact of probiotic administration on genes involved in homeostasis and immunity in goat blood. Following initial screening for infection, one-week post weaning, female SpanishXBoer goats were drenched daily with the recommended doses of FASTtrak microbial pack (Conklin Company Inc., Shakopee, MN) in 10 mL sterile water over an 8-week period. The control group were given sterile water. Blood samples were collected weekly. Total RNA was isolated from blood collected at the beginning of the study (week 0) and at the end of the study (week 8) using Tri-reagent and then reverse-transcribed to cDNA using the Ambion-Retroscript kit. Quantification of genes was performed in the CFX96 Probiotic treatment had no effect on body weight, body condition, fecal egg count and RNA concentration (p>0.05). Packed cell volume and FAMACHA scores were significantly improved by probiotic administration. Results from RT-PCR showed increased expression of genes in innate and adaptive immune response, cytokine and Wnt pathways in response to probiotics. Probiotics induced the expression of 32 genes involved in innate and adaptive immune response inflammatory cytokines, and 48 genes involved in the Wnt signaling pathway. This study provides evidence for a systematic effect of oral probiotic administration on expression of genes involved in immunity and homeostasis in goat blood.
Probiotics are viable microorganisms with beneficial health effects for humans and animals. They are formulated into many functional foods and animal feed. There is a growing research interest in the application and benefits of probiotics in ruminant production. Several recent studies have evaluated the potential of probiotics in animal nutrition and health. In this chapter, we have reviewed current research on the benefits of probiotics on gut microbial communities in ruminants and their impact on ruminant production, health and overall wellbeing.
Programs based on antibiotics are failing to control diseases due to increase in resistance of pathogens to antibiotics. Food safety, animal welfare and public health concerns have fueled interest in the use of plant-based alternatives. This study was conducted to evaluate the effect of a plant (Sericea Lespedeza, SL), and pathogen associated molecular patterns (PAMPs) (Lipopolysaccharide (LPS) and peptidoglycan (PGN)) on gene activation in ruminant blood. A water extract of SL, was used as a source of plant-derived tannins. Blood was collected from Holstein-Friesian cows (N = 4), Spanish × Boer goats (N = 4), St Croix sheep (N = 4) and incubated with 100 ng/mL of SL in the presence or absence of LPS or PGN. Samples maintained in Phosphate-buffered saline (PBS) served as negative control. The total protein concentration, WNT5a, and prostaglandin E2 in plasma were determined. Total RNA was isolated, reverse transcribed and Real time-PCR was performed using gene specific primers for TLR2, TLR4, WNT5a, and FZD. TLR2 and FZD were up-regulated in response to PAMPs. WNT5a and TLR4 genes were undetected in PAMP treated blood. SL regulated protein and prostaglandin concentration in all species. SL reduced PGE2 in sheep and cow blood. WNT5a was only secreted in LPS treated cow blood. Transcription and translation of genes involved in innate and adaptive immunity and the WNT signaling pathway in ruminant blood were responsive to diverse PAMPS, and can be modulated by SL. This suggests that dietary tannins may promote the health of ruminants. Further studies are needed to determine the significance of these changes in immune gene expression on ruminant health.
Probiotic supplements are beneficial for animal health and rumen function; and lipopolysaccharides (LPS) from gram negative bacteria have been associated with inflammatory diseases. In this study the transcriptional profile in whole blood collected from probiotics-treated cows was investigated in response to stimulation with lipopolysaccharides (LPS) in vitro. Microarray experiment was performed between LPS-treated and control samples using the Agilent one-color bovine v2 bovine (v2) 4x44K array slides. Global gene expression analysis identified 13,658 differentially expressed genes (fold change cutoff ≥ 2, P < 0.05), 3816 upregulated genes and 9842 downregulated genes in blood in response to LPS. Treatment with LPS resulted in increased expression of TLR4 (Fold change (FC) = 3.16) and transcription factor NFkB (FC = 5.4) and decreased the expression of genes including TLR1 (FC = − 2.54), TLR3 (FC = − 2.43), TLR10 (FC = − 3.88), NOD2 (FC = − 2.4), NOD1 (FC = − 2.45) and pro-inflammatory cytokine IL1B (− 3.27). The regulation of the genes involved in inflammation signaling pathway suggests that probiotics may stimulate the innate immune response of animal against parasitic and bacterial infections. We have provided a detailed description of the experimental design, microarray experiment and normalization and analysis of data which have been deposited into NCBI Gene Expression Omnibus (GEO): GSE75240.
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