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ABSTRACT: Spray-dried plasma (SDP) is a functional feed additive that has been established to improve performance and health of livestock. Understanding the effect of SDP in immune response modulation is essential to optimize its use for controlling Salmonella Enteritidis (SE) infection in chickens. This study was conducted to determine the levels of expression of selected cytokine genes in the ileum and cecal tonsil of SE-challenged broiler chicks. In a floor-pen housing, 320 broilers chicks were randomly assigned to 6 treatment groups: CX (unmedicated corn-soybean meal (SBM) basal without SDP), MX (unmedicated corn-SBM basal with antibiotic bacitracin methylene disalicylate (BMD) added at 0.055g/kg diet), PCX (unmedicated corn-SBM basal with SDP added at 30g/kg diet). Treatments SE, MSE, and PSE consisted of chicks inoculated with 7.46 × 108 CFU SE /mL at 1 d of age and given diets similar to CX, MX, and PCX, respectively. Samples of cecal tonsils and ileum were collected on d 3, 7 and 14 post infection for qRT-PCR analysis to determine the expression levels of interleukin (IL)-1β, interferon-γ (IFN-γ), IL-13, IL-17, IL-6, and transforming growth factor (TGF)-β genes. In the ceca tonsils, expression of IFN-γ was not affected by the interaction of Day and Treatment (P > 0.05). The level of the anti-inflammatory cytokine IL-13 was lower in MX and PCX on d 7 whereas high levels were expressed (P < 0.05) in MSE and PSE. In the ileum, expression of IL-17 and IFN-γ was significantly lower (P < 0.05) in PSE and MSE, but only PSE expressed lower IL-6 comparable to unchallenged treatments. On d 28 postchallenge, concentrations of anti-SE IgY and IL-6 protein were higher (P < 0.05) in the SE-challenged treatments compared to the unchallenged treatments. Overall, these results suggest that dietary SDP showed similar potency to BMD in modulating intestinal cytokine response against intestinal SE colonization in broiler chicks and therefore can be considered suitable alternative replacement for antibiotics in broiler production.

Due to increasing concerns about the contamination of animal food products with antibiotic-resistant bacteria and their byproducts, phytogenic feed additives in animal diets have been explored as antibiotic alternatives. In this study, we investigated the effect of ginger root extract (GRE), green tea extract (GTEC caffeinated and GTED decaffeinated), and onion peel combined (OPEC) on the activity of C. perfringens toxin genes and Eimeria tenella sporozoites. To this end, two Clostridium perfringens strains, CP19 and CP240 (Rollins Diagnostic Lab, Raleigh, NC, USA), were cultured (three replicates per treatment) as follows: without additives (Control), with Bacitracin Methylene Disalicylate (BMD), with GRE, with GTEC, with GTED, and, finally, with OPEC for 0, 2, 4, 6, 8, and 24 h. RNA was extracted to determine the expression of tpeL, alpha toxin (α-toxin), and NetB and we measured the protein concentration of NetB-positive C. perfringens toxin. Also, we evaluated the cytotoxic effect of green tea and ginger extracts on E. tenella sporozoites. Results show that phytogenic extracts, GRE, GTEC, and GTED, significantly reduced (p < 0.05) the level of expression of α-toxin gene compared to control; however, BMD treatment showed much less effect. Furthermore, NetB and tpeL encoding gene expression was significantly (p < 0.05) reduced by GRE and GTED, as well as BMD treatment, compared to the control. In contrast, GTEC treatment did not change the expression levels of these genes and was similar to control. With the CP240 strain, all the selected phytogenic extracts significantly reduced (p < 0.05) the expression of selected genes, except for OPEC, which was similar to control. GRE, GTEC, and GTED all reduced the viability of concentration of E. tenella sporozoites. Overall, our data show that these selected phytogenic extracts reduced the level of expression of toxin encoding genes associated with necrotic enteritis and decreased the viability of sporozoites which cause coccidiosis in broiler chicken.

This study was designed to assess the effect of ginger root extract (GRE) supplementation on the oxidative status and intestinal mucosal development in broiler chickens for 6 weeks. Day-old chicks (Ross 708 strain, n = 432) were distributed into six treatments with six replicate of twelve birds each: Negative CON (basal), MX (basal diet + bacitracin methylene disalicylate (BMD) 0.055 g/kg diet), GRE-1 (basal diet + 0.375% GRE), GRE-2 (basal diet + 0.75% GRE), GRE-3 (basal diet + 1.5% GRE), GRE-4 (basal diet + 3% GRE). Growth indices, goblets cell count, mucin (MUC2) in ileum tissue, antioxidant (SOD, CAT, and GPX) in ileum and liver, biological antioxidant potential (BAP), and reactive oxygen metabolite level in blood and intestinal villi measurement were determined. Body weight (BW) was highest (p < 0.05) in all groups except GRE-4, body weight gain (BWG) was best in GRE-1, while FCR was least in all groups except GRE-4. Optimum MUC2 gene expression, SOD, CAT, blood antioxidants, and intestinal morphometric values were observed in GRE-3. The inclusion of ginger root extract up to 1.5% improved growth and reduced oxidative stress while enhancing mucosal development in broiler chicks.

ABSTRACT: Ginger contains bioactive compounds that possess anti-inflammatory and antimicrobial properties. In this study, 432-day-old Ross 708 broiler male chicks were randomly allocated to 6 dietary treatments to investigate the effect of ginger root extract (GRE) on immunocompetence and growth performance to 6 wk of age. Treatment 1 (CON) consisted of chicks fed a corn–soybean meal (SBM), a base diet without GRE. Treatment 2 (MX) chicks were given basal diets containing bacitracin methylene disalicylate (BMD) at 0.055 g/kg. Treatments 3 (GRE-0.375%), 4 (GRE-0.75%), 5 (GRE-1.5%), and 6 (GRE-3%) were fed similar diet to control with GRE supplemented at 0.375%, 0.75%, 1.5%, and 3%, respectively. Moreover, HPLC analysis of GRE was carried out to determine the concentration of bioactive compounds found in GRE. Each treatment consisted of 6 replicate pens with 12 chicks/pen. Bodyweight (BW) and feed conversion ratio (FCR) were recorded. Results show that the concentration of bioactive compounds increased with increasing GRE supplementation. Likewise, dietary GRE supplementation did not have any detrimental effect on growth performance parameters up to 1.5%, as values for BWG was not different from CON and MX; however, 3% GRE had the poorest FCR and a lower BWG as compared to other treatments. On d 27 and d 41, fecal and cecal concentrations of total bacteria count (TBC), Escherichia coli, Lactobacillus spp., and Bifidobacterium spp enumerated using selective plating media showed that GRE supplementation significantly reduced (P < 0.05) the amount of TBC and E. coli but increased the number of beneficial microorganisms such as Lactobacillus spp. and Bifidobacterium spp. On d 20, no significant differences were observed (P > 0.05) among all treatments for antibody titer against Newcastle disease virus and total IgY antibodies; however, on d 27, GRE-0.75% had the highest value for both immune indicators and was not different from MX. Dietary supplementation of GRE up to 1.5% enhanced the immune system and suppressed E. coli while promoting the growth of healthy bacteria, without any detrimental effect on growth performance.

Semen preservation involves lengthening sperm’s fertile lifespan without any detrimental effects on its biochemical, functional, and ultrastructural properties. Liquid storage at 4 °C is a ram sperm preservation method. However, this method of storage causes irreversible damage due to cold shocks, osmotic stresses, oxidative stresses, and reductions in sperm metabolism. The present study aims to investigate whether the supplementation of mitochonic acid 5 (MA-5) in a sperm extender could improve chilled ram sperm quality and elucidate its mechanism of action. Ram sperm were diluted with a tris-citrate-glucose extender containing different concentrations of MA-5 (0, 0.1, 1, 10, and 100 nM) and stored at 4 °C for up to 48 h. Sperm motility, membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) level, ATP content, and the expression of NADPH dehydrogenase subunits 1 (MT-ND1) and NADPH dehydrogenase subunits 6 (MT-ND6) were evaluated. It was observed that compared to the control, the 10 nM MA-5 treatment significantly (p < 0.05) increased total motility (82 ± 3.5% vs. 76 ± 5.9%), progressive motility (67.6 ± 8.2% vs. 51 ± 8.3%), and other parameters (straight-line velocity (VSL), average path velocity (VAP), and curvilinear velocity (VCL)). In addition, 10 nM MA-5 supplementation also improved ram sperm membrane integrity and acrosomal integrity as well increased mitochondrial membrane potential (51.1 ± 0.7% vs. 37.7 ± 1.3%), reduced ROS levels, and elevated adenosine triphosphate (ATP) contents. Furthermore, a Western blot analysis demonstrated that the addition of MA-5 significantly (p < 0.05) increased the expression of MT-ND1 and MT-ND6 proteins in ram sperm, with the 10 nM MA-5 treatment resulting in the highest expression level. These results suggest that MA-5 improves ram sperm quality by maintaining high sperm mitochondrial function during liquid storage at 4 °C.

Sperm motility is an important factor in the migration of sperm from the uterus to the oviduct. During sperm preservation in vitro, sperm generates excessive ROS that damages its function. This study aims to investigate whether the addition of pyrroloquinoline quinone (PQQ) to the diluted medium could improve chilled ram sperm quality, and then elucidates the mechanism. Ram semen was diluted with Tris-citric acid-glucose (TCG) medium containing different doses of PQQ (0 nM, 10 nM, 100 nM, 1000 nM, 10,000 nM), and stored at 4 °C. Sperm motility patterns, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) levels, malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, and ATP levels were measured after preservation. Furthermore, the expressions of NADH dehydrogenase 1 (MT-ND1) and NADH dehydrogenase 6 (MT-ND6) in sperm were also detected by western blotting. In addition, sperm capacitation and the ability of sperm to bind to the zona pellucina were also evaluated. It was observed that the addition of PQQ significantly (p < 0.05) improved ram sperm motility, membrane integrity, and acrosome integrity during preservation. The percentage of sperm with high mitochondrial membrane potential in the PQQ treatment group was much higher than that in the control. In addition, supplementation of PQQ also decreased the sperm MDA and ROS levels, while increasing ATP levels. Interestingly, the levels of MT-ND1 and MT-ND6 protein in sperm treated with PQQ were also higher than that of the control. Furthermore, the addition of 100 nM PQQ to the medium decreased ROS damage in MT-ND1 and MT-ND6 proteins. The addition of 100 nM PQQ significantly (p < 0.05) increased protein tyrosine phosphorylation in ram sperm after induced capacitation. Furthermore, the value of the sperm–zona pellucida binding capacity in the 100 nM PQQ treatment group was also much higher than that of the control. Overall, during chilled ram- sperm preservation, PQQ protected ram sperm quality by quenching the ROS levels to reduce ROS damage and maintain sperm mitochondrial function, and preserved the sperm’s high ability of fertilization.

Cryopreservation generates a substantial quantity of ROS in semen, leading to a decline in sperm quality and fertilization capacity. The objective of this study was to investigate the effects of resveratrol and its optimal concentration on ram sperm quality after cryopreservation. Ram semen was diluted with a freezing medium containing different concentrations of resveratrol (0, 25, 50, 75, and 100 μM). After thawing, various sperm parameters such as total motility, progressive motility, acrosome integrity, plasma membrane integrity, mitochondrial membrane potential, glutathione (GSH) content, glutathione synthase (GPx) activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, lipid peroxidation (LPO) content, malondialdehyde (MDA) content, ROS level, SIRT1 level, DNA oxidative damage, and AMPK phosphorylation level were assessed. In addition, post-thaw sperm apoptosis was evaluated. Comparatively, the addition of resveratrol up to 75 μM significantly improved the sperm motility and sperm parameters of cryopreserved ram sperm. Specifically, 50 μM resveratrol demonstrated a notable enhancement in acrosome and plasma membrane integrity, antioxidant capacity, mitochondrial membrane potential, adenosine triphosphate (ATP) content, SIRT1 level, and AMPK phosphorylation levels compared to the control group (p < 0.05). It also significantly (p < 0.05) reduced the oxidative damage to sperm DNA. However, detrimental effects of resveratrol were observed at a concentration of 100 μM resveratrol. In conclusion, the addition of 50 μM resveratrol to the cryopreservation solution is optimal for enhancing the quality of cryopreserved ram sperm.

Spray-dried plasma (SDP) contain a variety of functional proteins that play an immunomodulatory role. To evaluate the potential of SDP to stimulate the immune system, day-old Ross 708 male broiler chicks (200) were allocated randomly to five dietary treatments. Treatment 1 (CX) comprised chicks fed basal unmedicated corn–soybean meal (SBM) without the addition of SDP. Treatment 2 (MX) includes chicks fed unmedicated corn–SBM basal containing Bacitracin methylene disalicylate (BMD) at 0.055 g/kg diet. Treatments 3 (SDP1), 4 (SDP2), and 5 (SDP3) contained chicks given unmedicated corn–SBM basal, into which SDP was included at 10, 20, and 30 g/kg diet, respectively. On d 7, 14, and 21, chicks’ body weight and FCR were calculated. Additionally, leucocyte counts, oxidative status, and IgY concentrations were determined in blood. On d 23, fecal populations of selected indicator bacteria species were determined. Results showed that FCR for SP3 was superior (p < 0.05) to other treatments. Likewise, heterophil numbers decreased in MX and SDP treatments compared to CX. Circulating IgY concentration was higher for SDP dietary treatments (p < 0.05) compared to MX. In conclusion, dietary SDP at 30 g/kg enhanced immune surveillance by increasing circulating IgY levels, maintaining a normal oxidative state, and increasing gut Bifidobacteria, thereby improving chick growth performance.