Analysis of the immunomodulatory properties of mycobacterium cell wall fraction on the cytokine production of peripheral blood mononuclear cells of healthy dogs.

BACKGROUND: Mycobacterium cell wall fraction (MCWF) is derived from nonpathogenic Mycobacterium phlei and is used as an immunomodulatory compound in clinical practice, yet its mode-of-action requires further research. OBJECTIVE: To evaluate the host response to MCWF in canine peripheral blood mononuclear cells (PBMCs) by using enzyme-linked immunosorbent assays (ELISA) and quantitative reverse transcription (qRT)-PCR for assessment of cytokines. ANIMALS: Eight healthy Labrador retrievers. MATERIALS AND METHODS: PBMCs were isolated from whole blood using density centrifugation. The cells were cultured with different concentrations of MCWF or a potent stimulator of cytokine production, phorbol 12-myristate 13-acetate/ionomycin, or left in cell culture medium for 24, 48 and 72 h. Cytokines were measured by ELISA for interleukin (IL)-4, IL-10 and interferon-gamma (IFN-γ), and by qRT-PCR for IL-4, IL-10, IL-13, IFN-γ, tumour necrosis factor alpha (TNF-α) and transforming growth factor-beta. RESULTS: A significant increase of IL-10 messenger ribonucleic acid (mRNA) was detected at all time points for all concentrations of MCWF (p < 0.05). Protein analysis reflected this finding, with a maximum IL-10 concentration of 300.6 ± 38.3 µg/mL. Compared to the negative control, post-stimulation elevation of IFN-γ mRNA was noted at 24 h with all concentrations of MCWF (p < 0.01), and TNF-α mRNA was increased for 0.5 µg/dL MCWF only at 72 h (p < 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: MCWF stimulation of PBMCs results in the elevation of both proinflammatory and regulatory cytokine mRNA. Further research into the role of MCWF as a systemically administered regulatory immunomodulator or adjuvant to allergen-specific immunotherapy should be considered.

An evaluation of the thermoregulatory potential of in ovo delivered bioactive substances (probiotic, folic acid, and essential oil) in broiler chickens.

Mitigating the negative effects of heat stress (HS) is a critical challenge for the global poultry industry. This study evaluated the thermoregulatory potential of 3 in ovo delivered bioactive substances using selected gut health parameters. Eggs were incubated and allotted to 5 groups, and respective bioactive substances delivered. These groups included-the noninjected, in ovo saline, in ovo folic acid (FA), in ovo probiotics (P), and in ovo essential oil (EO). At hatch, chicks were assigned to 5 new posthatch treatment combinations, including A) Negative control (NC)-noninjected eggs offered a basal corn-wheat-soybean diet, B) Antibiotics-NC + 0.05% bacitracin methylene disalicylate, C) In ovo FA-eggs injected with FA + NC diet, D) In ovo probiotics-eggs injected with probiotics + NC diet, E) In ovo + in-water EO-eggs injected with EO and supplied EO via drinking water + NC diet. Birds were raised for 28 d in 8 replicate cages/treatment (6 birds/cage) and exposed to either a thermoneutral (24°C ± 0.2) or HS challenge (31°C) condition from d 21 to d 28. The in ovo delivered FA and EO treatments reduced (P < 0.001) hatchability by at least 26% compared to NC. Induced HS reduced (P < 0.001) total plasma protein, total antioxidant capacity, and villus width in the duodenum and jejunum. Independent of HS and compared to NC, the in ovo + in-water EO treatment resulted in (P < 0.05) at least a 15% increase in villus height: crypt depth across the 3 gut sections. The in ovo + in-water EO treatment also increased the relative mRNA expression of intestinal barrier-related genes (Claudin1,3,4, Occludin, Zona occludens-2, and Mucin 2). Under HS, the in ovo + in-water EO treatment recorded a 3.5-fold upregulation of amino acid transporter gene (SLC1A1), compared to NC. Subject to further hatchability optimization, the in ovo + in-water delivery of EO show potential to afford broiler chicken thermotolerance.

Microbiota and Transcriptomic Effects of an Essential Oil Blend and Its Delivery Route Compared to an Antibiotic Growth Promoter in Broiler Chickens.

This study evaluated the effect of the delivery of a commercial essential oil blend containing the phytonutrients star anise, cinnamon, rosemary, and thyme oil (via different routes) on broiler chickens' ileal and ceca microbiota and liver transcriptome compared to an antibiotic growth promoter. Eggs were incubated and allocated into three groups: non-injected, in ovo saline, and in ovo essential oil. On day 18 of incubation, 0.2 mL of essential oil in saline (dilution ratio of 2:1) or saline alone was injected into the amnion. At hatch, chicks were assigned to post-hatch treatment combinations: (A) a negative control (corn-wheat-soybean diet), (B) in-feed antibiotics, (C) in-water essential oil (250 mL/1000 L of drinking water), (D) in ovo saline, (E) in ovo essential oil, and (F) in ovo essential oil plus in-water essential oil in eight replicate cages (six birds/cage) and raised for 28 days. On days 21 and 28, one and two birds per cage were slaughtered, respectively, to collect gut content and liver tissues for further analysis. Alpha and beta diversity differed significantly between ileal and ceca samples but not between treatment groups. In-feed antibiotic treatment significantly increased the proportion of specific bacteria in the family Lachnospiraceae while reducing the proportion of bacteria in the genus Christensenellaceae in the ceca, compared to other treatments. Sex-controlled differential expression of genes related to cell signaling and tight junctions were recorded. This study provides data that could guide the use of these feed additives and a foundation for further research.

Essential Oil Delivery Route: Effect on Broiler Chicken's Growth Performance, Blood Biochemistry, Intestinal Morphology, Immune, and Antioxidant Status.

This study evaluated the effect of an essential oil blend and its delivery routes on broiler chicken growth performance, blood biochemistry, intestinal morphology, and immune and antioxidant status. Eggs were incubated and allotted to 3 groups: non-injected group, in ovo saline group, and in ovo essential oil group. On day 18 of incubation, essential oil in saline or saline alone was injected into the amnion. At hatch, chicks were assigned to post-hatch treatment combinations (1) in ovo essential oil + in-water essential oil (in ovo + in-water EO); (2) in ovo essential oil (in ovo EO); (3) in ovo saline; (4) in-water essential oil; (5) in-feed antibiotics (Bacitracin methylene disalicylate) and (6) a negative control (NC; corn-wheat-soybean diet) in 8 replicate cages (6 birds/cage) and raised for 28 day. The in ovo EO group reduced (p < 0.05) chick length and hatchability, all groups recorded no difference in growth performance at 0-28 day. The in ovo + in-water EO treatment reduced (p < 0.05) blood creatine kinase and aspartate aminotransferase levels whilst increasing (p < 0.05) total antioxidant capacity in birds. The in ovo + in-water delivery of EO might represent a potential antibiotic reduction strategy for the poultry industry but more research is needed to address the concern of reduced hatchability.