Agaro-oligosaccharides mitigate deoxynivalenol-induced intestinal inflammation by regulating gut microbiota and enhancing intestinal barrier function in mice.

Agarose-derived agaro-oligosaccharides (AgaroS) have been extensively studied in terms of structures and bioactivities; they reportedly possess antioxidant and anti-inflammatory activities that maintain intestinal homeostasis and host health. However, the protective effects of AgaroS on deoxynivalenol (DON)-induced intestinal dysfunction remain unclear. We investigated the effects of AgaroS on DON-induced intestinal dysfunction in mice and explored the underlying protective mechanisms. In total, 32 mice were randomly allocated to four treatments (n = 8 each) for 28 days. From day 1 to day 21, the control (CON) and DON groups received oral phosphate-buffered saline (200 µL per day); the AgaroS and AgaroS + DON groups received 200 mg AgaroS per kg body weight once daily by orogastric gavage. Experimental intestinal injury was induced by adding DON (4.8 mg per kg body weight) via gavage from day 21 to day 28. Phosphate-buffered saline was administered once daily by gavage in the CON and AgaroS groups. Herein, AgaroS supplementation led to a higher final body weight and smaller body weight loss and a lower concentration of plasma inflammatory cytokines, compared with the DON group. The DON group showed a significantly reduced ileal villus height and villus height/crypt depth, compared with the CON and AgaroS + DON groups. However, AgaroS supplementation improved DON-induced intestinal injury in mice. Compared with the DON group, ileal and colonic protein expression levels of claudin, occludin, Ki67, and mucin2 were significantly higher in the AgaroS supplementation group. Colonic levels of the anti-inflammatory cytokine IL-1ß tended to be higher in the DON group than in the AgaroS + DON group. AgaroS altered the gut microbiota composition, accompanied by increased production of short-chain fatty acids in mice. In conclusion, our findings highlight a promising anti-mycotoxin approach whereby AgaroS alleviate DON-induced intestinal inflammation by modulating intestinal barrier functional integrity and gut microbiota in mice.

Effects of dietary methionine supplementation on the growth performance, immune responses, antioxidant capacity, and subsequent development of layer chicks.

Deficiencies or excesses of dietary amino acids, and especially of methionine (Met), in laying hens can lead to abnormal protein anabolism and oxidative stress, which affect methylation and cause cellular dysfunction. This study investigated the effects of dietary methionine (Met) levels on growth performance, metabolism, immune response, antioxidant capacity, and the subsequent development of laying hens. A total of 384 healthy 1-day-old Hyline Grey chicks of similar body weight were randomly allocated to be fed diets containing 0.31%, 0.38%, 0.43% (control group), or 0.54% Met for 6 wk, with 6 replicates of 16 chicks in each. The growth performance of the chicks was then followed until 20 wk old. The results showed dietary supplementation with 0.43% or 0.54% Met significantly increased their mean daily body weight gain, final weight, and Met intake. However, the feed:gain (F/G) decreased linearly with increasing Met supplementation, from 0.31 to 0.54% Met. Met supplementation increased the serum albumin, IgM, and total glutathione concentrations of 14-day-old chicks. In contrast, the serum alkaline phosphatase activity and hydroxyl radical concentration tended to decrease with increasing Met supplementation. In addition, the highest serum concentrations of IL-10, T-SOD, and GSH-PX were in the 0.54% Met-fed group. At 42 d of age, the serum ALB, IL-10, T-SOD, GSH-PX, T-AOC, and T-GSH were correlated with dietary Met levels. Finally, Met supplementation reduced the serum concentrations of ALP, IL-1ß, IgA, IgG, hydrogen peroxide, and hydroxyl radicals. Thus, the inclusion of 0.43% or 0.54% Met in the diet helps chicks achieve superior performance during the brooding period and subsequently. In conclusion, Met doses of 0.43 to 0.54% could enhance the growth performance, protein utilization efficiency, antioxidant capacity, and immune responses of layer chicks, and to promote more desirable subsequent development during the brooding period.

Using low-protein diet in egg production for win-win of productivity and environmental benefits should be prudent: Evidence from pilot test.

A shortage of feed protein resources restricts poultry productivity. Key strategies to alleviate this problem include improvements to the structure of the gut microbiota by the appropriate intake of high-quality protein, improvements to the comprehensive protein utilization rate, and reducing the consumption of protein raw materials. In addition, damage to the environment caused by nitrogen emissions needs to be reduced. The aim of the study was to evaluate the effects of dietary protein levels on laying performance, host metabolism, ovarian health, nitrogen emissions, and the gut microbial structure and function of laying hens. In total, 360 hens at the age of 38 weeks were randomly allotted four treatments. Each of the groups consisted of nine replicates, with 10 birds per replicate, used for 12 weeks of study. Dietary protein levels of the four groups were 13.85 %, 14.41 %, 15.63 %, and 16.30 %. Results revealed that, compared with the 13.85 % crude protein (CP) group, the 15.63 % CP group experienced significantly enhanced final body weight, average daily gain, egg production, and egg mass. Compared with the 16.30 % CP group, the other groups' serum concentrations of immunoglobulin G (IgG) and immunoglobulin M (IgM) were significantly reduced. Compared with the 16.30 % CP group, the 13.85 % and 15.63 % groups had increased CP utilization rates but reduced nitrogen emission rate, and daily per egg and per kilogram egg nitrogen emissions rose with increased dietary protein levels. Compared to the 13.85 % and 14.41 % CP groups, the 16.30 % CP group exhibited a significant increase in the expression of genes related to amino acids and carbohydrate metabolic pathways. According to the linear discriminant analysis effect size diagram, the predominant bacteria in the 15.63 % CP group (e.g., Subdoligranulum, and Ruminococcaceae_UCG-013) were significantly related to CP utilization. The results of this study emphasize that production performance is significantly reduced when protein levels are too low, whereas too high protein levels lead to gut microbiota imbalance and a reduction in the utilization efficiency of nutrients. Therefore, on the premise of ensuring the health of hens, the structure of the gut microbiota can be improved by appropriately reducing protein levels, which helps to balance the relationships among host health, productivity, resources, and the environment.

Yeast polysaccharide mitigated oxidative injury in broilers induced by mixed mycotoxins via regulating intestinal mucosal oxidative stress and hepatic metabolic enzymes.

This study was aimed to investigate the effects of yeast polysaccharides (YPS) on growth performance, intestinal health, and aflatoxin metabolism in livers of broilers fed diets naturally contaminated with mixed mycotoxins (MYCO). A total of 480 one-day-old Arbor Acre male broilers were randomly allocated into a 2 × 3 factorial arrangement of treatments (8 replicates with 10 birds per replicate) for 6 wk to assess the effects of 3 levels of YPS (0, 1, or 2 g/kg) on the broilers fed diets contaminated with or without MYCO (95 µg/kg aflatoxin B1, 1.5 mg/kg deoxynivalenol, and 490 µg/kg zearalenone). Results showed that mycotoxins contaminated diets led to significant increments in serum malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels, mRNA expressions of TLR4 and 4EBP1 associated with oxidative stress, mRNA expressions of CYP1A1, CYP1A2, CYP2A6, and CYP3A4 associated with hepatic phase Ⅰ metabolizing enzymes, mRNA expressions of p53 associated with hepatic mitochondrial apoptosis, and AFB1 residues in the liver (P < 0.05); meanwhile dietary MYCO decreased the jejunal villus height (VH), villus height/crypt depth (VH/CD), the activity of serum total antioxidant capacity (T-AOC), mRNA expressions of jejunal HIF-1α, HMOX, and XDH associated with oxidative stress, mRNA expressions of jejunal CLDN1, ZO1, and ZO2, and mRNA expression of GST associated with hepatic phase Ⅱ metabolizing enzymes of broilers (P < 0.05). Notably, the adverse effects induced by MYCO on broilers were mitigated by supplementation with YPS. Dietary YPS supplementation reduced the concentrations of serum MDA and 8-OHdG, jejunal CD, mRNA expression of jejunal TLR2, and 4EBP1, hepatic CYP1A2, and p53, and the AFB1 residues in the liver (P < 0.05), and elevated the serum T-AOC and SOD, jejunal VH, and VH/CD, and mRNA expression of jejunal XDH, hepatic GST of broilers (P < 0.05). There were significant interactions between MYCO and YPS levels on the growth performance (BW, ADFI, ADG, and F/G) at d 1 to 21, d 22 to 42, and d 1 to 42, serum GSH-Px activity, and mRNA expression of jejunal CLDN2 and hepatic ras of broilers (P < 0.05). In contrast with MYCO group, the addition of YPS increased BW, ADFI, and ADG, the serum GSH-Px activity (14.31%-46.92%), mRNA levels of jejunal CLDN2 (94.39%-103.02%), decreased F/G, and mRNA levels of hepatic ras (57.83%-63.62%) of broilers (P < 0.05). In conclusion, dietary supplements with YPS protected broilers from mixed mycotoxins toxicities meanwhile keeping normal performance of broilers, presumably via reducing intestinal oxidative stress, protecting intestinal structural integrity, and improving hepatic metabolic enzymes to minimize the AFB1 residue in the liver and enhance the performance of broilers.

Porcine bile acids promote the utilization of fat and vitamin A under low-fat diets.

Fat-soluble vitamin malabsorption may occur due to low dietary fat content, even in the presence of an adequate supply of fat-soluble vitamins. Bile acids (BAs) have been confirmed as emulsifiers to promote fat absorption in high-fat diets. However, there are no direct evidence of exogenous BAs promoting the utilization of fat-soluble vitamins associated with fat absorption in vitro and in vivo. Therefore, we chose laying hens as model animals, as their diet usually does not contain much fat, to expand the study of BAs. BAs were investigated in vitro for emulsification, simulated intestinal digestion, and release rate of fat-soluble vitamins. Subsequently, a total of 450 healthy 45-week-old Hy-Line Gray laying hens were chosen for an 84-day feeding trial. They were divided into five treatments, feeding diets supplemented with 0, 30, 60, 90, and 120 mg/kg BAs, respectively. No extra fat was added to the basic diet (crude fat was 3.23%). In vitro, BAs effectively emulsified the water-oil interface. Moreover, BAs promoted the hydrolysis of fat by lipase to release more fatty acids. Although BAs increased the release rates of vitamins A, D, and E from vegetable oils, BAs improved for the digestion of vitamin A more effectively. Dietary supplementation of 60 mg/kg BAs in laying hens markedly improved the laying performance. The total number of follicles in ovaries increased in 30 and 60 mg/kg BAs groups. Both the crude fat and total energy utilization rates of BAs groups were improved. Lipase and lipoprotein lipase activities were enhanced in the small intestine in 60, 90, and 120 mg/kg BAs groups. Furthermore, we observed an increase in vitamin A content in the liver and serum of laying hens in the 60, 90, and 120 mg/kg BAs groups. The serum IgA content in the 90 and 120 mg/kg BAs groups was significantly improved. A decrease in serum malondialdehyde levels and an increase in glutathione peroxidase activity were also observed in BAs groups. The present study concluded that BAs promoted the absorption of vitamin A by promoting the absorption of fat even under low-fat diets, thereupon improving the reproduction and health of model animals.

Dietary selenium sources alleviate immune challenge induced by Salmonella Enteritidis potentially through improving the host immune response and gut microbiota in laying hens.

The aim of this study was to evaluate the effects of different selenium (Se) sources on the immune responses and gut microbiota of laying hens challenged with Salmonella enteritidis (S. Enteritidis). A total of 240 45-week-old layers were randomly divided into eight groups with six replicates per group according to a 4 × 2 factorial design, including a blank diet without Se supplementation (CON group) and three diets with 0.3 mg/kg Se supplementation from sodium selenite (IS group), yeast Se (YS group), and selenium-enriched yeast culture (SYC group), respectively. After 8 weeks of feeding, half of them were orally challenged with 1.0 ml suspension of 109 colony-forming units per milliliter of S. Enteritidis daily for 3 days. The serum was collected on days 3, 7, and 14, and the cecum content was collected on day 14 after challenge. There was no significant difference in laying performance among the eight groups before challenge. The S. Enteritidis challenge significantly decreased the laying performance, egg quality, GSH-Px, IgG, and IgM and increased the ratio of feed and egg, malondialdehyde (MDA), Salmonella-specific antibody (SA) titers, IL-6, IL-2, IL-1ß, and INF-γ. However, SYC increased the level of GSH-Px and IgG and decreased IL-6, while YS decreased the level of IL-2 and IL-1ß. What is more, Se supplementation decreased the SA titers to varying degrees and reduced the inflammatory cell infiltration in the lamina propria caused by S. Enteritidis infection. In addition, the S. Enteritidis challenge disrupted the intestinal flora balance by reducing the abundance of the genera Clostridium innocuum, Lachnospiraceae, and Bifidobacterium and increasing the genera Butyricimonas and Brachyspira, while Se supplementation increased the gut microbial alpha diversity whether challenged or not. Under the S. Enteritidis challenge condition, the alteration of microbial composition by the administration of different Se sources mainly manifested as IS increased the relative abundance of the genera Lachnospiraceae and Christensenellaceae, YS increased the relative abundance of the genera Megamonas and Sphingomonas, and SYC increased the genera Fusobacterium and Lactococcus. The alteration of gut microbial composition had a close relationship with antioxidant or immune response. To summarize, different Se sources can improve the egg quality of layers challenged by S. Enteritidis that involves elevating the immunity level and regulating the intestinal microbiota.

R- Is Superior to S-Form of α-Lipoic Acid in Anti-Inflammatory and Antioxidant Effects in Laying Hens.

The development of single enantiomers with high efficiency and low toxic activity has become a hot spot for the development and application of drugs and active additives. The aim of the present study was to investigate the effectiveness of the application of α-lipoic acid with a different optical rotation to alleviate the inflammation response and oxidative stress induced by oxidized fish oil in laying hens. Sixty-four 124-week-old Peking Red laying hens were randomly allocated to four groups with eight replicates of two birds each. The normal group was fed basal diets supplemented with 1% fresh fish oil (FO), and the oxidative stress model group was constructed with diets supplemented with 1% oxidized fish oil (OFO). The two treatment groups were the S-form of the α-lipoic acid model with 1% oxidized fish oil (OFO + S-LA) and the R-form of the α-lipoic acid model with 1% oxidized fish oil (OFO + R-LA) added at 100 mg/kg, respectively. Herein, these results were evaluated by the breeding performance, immunoglobulin, immune response, estrogen secretion, antioxidant factors of the serum and oviduct, and pathological observation of the uterus part of the oviduct. From the results, diets supplemented with oxidized fish oil can be relatively successful in constructing a model of inflammation and oxidative stress. The OFO group significantly increased the levels of the serum inflammatory factor (TNF-α, IL-1ß, IL-6, and IFN-γ) and the oxidative factor MDA and decreased the activity of the antioxidant enzyme (T-AOC, T-SOD, GSH-Px, GSH, and CAT) in the oviduct. The addition of both S-LA and R-LA significantly reduced the levels of serum inflammatory factors (TNF-α, IL-1ß, IL-6, and IFN-γ), increased the activity of antioxidant indexes (T-AOC, T-SOD, GSH-Px, GSH, and CAT), and decreased the MDA contents in the serum and oviduct. Meanwhile, the supplementation of S-LA and R-LA also mitigated the negative effects of the OFO on the immunoglobulins (IgA and IgM) and serum hormone levels (P and E2). In addition, it was worth noting that the R-LA was significantly more effective than the S-LA in some inflammatory (IL-1ß) and antioxidant indices (T-SOD, GSH, and CAT). Above all, both S-LA and R-LA can alleviate the inflammation and oxidative damage caused by oxidative stress in aged laying hens, and R-LA is more effective than S-LA. Thus, these findings will provide basic data for the potential development of α-lipoic acid as a chiral dietary additive for laying hens.

Vitamin D3 Nutritional Status Affects Gut Health of Salmonella-Challenged Laying Hens.

Salmonella enterica serovar Enteritidis (SE) is one of the most common pathogens associated with poultry health and foodborne Salmonellosis worldwide. The gut plays a pivotal role in inhibiting SE transintestinal transmission and contaminating poultry products. The nutritional status of vitamin D (VD) is involved in gut health apart from bone health. However, the impact of VD3 nutritional status on the gut health of Salmonella-challenged hens is rarely investigated. This study investigated the impact and possible mechanisms of VD3 nutritional status on the gut health of hens challenged with SE. Hens were fed basal diets with either 0 (deficient) or 3000 IU (sufficient) VD3/kg of diet, respectively. After 10 weeks of feeding, half of the hens were orally inoculated with either SE (1 × 109 CFU /bird). Results indicated that VD3 sufficiency reversed the disruptive effects on the laying performance of hens caused by Salmonella challenge or VD3 insufficiency by promoting VD3 metabolism. In addition, VD3 sufficiency ameliorated gut injury induced by either Salmonella or VD3 deficiency, shown by reducing Salmonella load and histopathological scores, suppressing TLR4-mediated inflammatory responses, and increasing expression of TJs along with decreasing pro-apoptotic protein expression and the number of TUNEL-positive cells in the jejunum. Besides, VD3 enriched the abundance of probiotics, such as Lactobacillus and Bacilli, and restored the balance of gut microflora. Collectively, dietary VD3 sufficient supplementation could alleviate Salmonella or VD3 deficiency-induced intestinal damage of hens via modulating intestinal immune, barrier function, apoptosis along with gut microbiota composition, revealing that VD3 could act as a novel nutritional strategy defending Salmonella invasion in hens.

Effect of Methionine Deficiency on the Growth Performance, Serum Amino Acids Concentrations, Gut Microbiota and Subsequent Laying Performance of Layer Chicks.

This study was conducted to investigate the effect of methionine (Met) deficiency in the rearing period on the growth performance, amino acids metabolism, intestinal development and gut microbiome of egg-laying chicks and the continuous effects on the performance, egg quality, and serum amino acids metabolism of the subsequent development process. Three hundred sixty one-day-old chicks were randomly divided into two groups and fed on a basal diet (NC group, Met 0.46%) and Met deficiency diet (Met- group, Met 0.27%). Each group included six replicates with 30 chicks per replicate. The trial lasted 6 weeks (0-6 weeks), both groups were fed the same basal diet which met the needs of Met during the observation period (7-24 weeks). Results showed that Met deficiency significantly decreased (P < 0.05) body weight (BW), average daily weight gain (ADG), average daily feed intake (ADFI) and tibia length (TL) compared to the NC group during the trial period (0-6 weeks). Also, Met deficiency dramatically increased (P < 0.05) feed conversion ratio (FCR) during the trial and observation period (7-24 weeks). In addition, during the observation period, the BW and ADG were decreased (P < 0.05) in the Met- group. Moreover, Met- group decreased (P < 0.05) villi height and villi height/crypt depth ratio in jejunum at 6th weeks. In addition, the concentrations of serum main free amino acids (FAA) in the Met- group were significantly increased (P < 0.05) at 6th weeks, while were decreased at 16th weeks. Based on the α-diversity and PCoA analysis in ß-diversity, there were no significant differences in the cecal microbial composition between NC and Met- groups. However, the LEfSe analysis revealed that differential genera were enriched in the NC or Met- groups. The Haugh unit, shell thickness and egg production in the Met- group were significantly lower (P < 0.05) than in the NC group. In conclusion, these results revealed that dietary supplementation of appropriate Met could substantially improve the growth performance, host amino acid metabolism and intestinal development and continuously improve the laying performance and thus boost the health of growing hens.

Phosphorus Restriction in Brooding Stage Has Continuous Effects on Growth Performance and Early Laying Performance of Layers.

This study evaluated the effects of phosphorus restriction in the brooding stage and subsequent recovery on growth performance, tibia development and early laying performance of layers. 360 one-day-old hens were randomly divided into 4 groups with 6 replicates and 15 chicks per replicate. Chicks were fed diets containing 0.13% (L), 0.29% (M), 0.45% (N), 0.59% (H) non-phytate phosphorus (nPP) from 1 to 8 weeks of age. From 9 to 20 weeks of age, the L and N group were divided into two groups fed normal level phosphorus (n, 0.39% nPP) and high-level phosphorus (h, 0.45% nPP) separately, then all the birds were fed a normal diet (0.39% nPP) from 21 to 26 weeks of age. Four treatments were tested: Ln, Lh, Nn, and Nh. The lower body weight, average daily feed intake, tibia length and daily tibial increment were observed in the L group (p < 0.05) and the ratio of feed to gain was significantly increased in the L group at 8 weeks of age (p < 0.05). In addition, the fresh and degreased tibia weight, bone ash, Ca content in the tibia and P content in the ash and tibia were significantly decreased in the L group at 8 weeks of age (p < 0.05). After compensatory processes, there was no significant difference in tibia characters; however, body weight in the Ln group was significantly lower than in the Nn group (p < 0.05) and was significantly lower in the Lh group than the Nn group (p < 0.01) and Nh group (p < 0.05). In addition, the laying rate and average daily egg mass in the Lh group were lower than Nn and Nh (p < 0.05). In conclusion, severe dietary phosphorus restriction impaired growth performance and bone mineralization in the brooding stage. Subsequent phosphorus supplementation could not alleviate this adverse effect on body weight, which continued to affect egg production. These findings give a foundation and new perspective on a low phosphorus feeding strategy in layer production.

Vitamin K alleviates bone calcium loss caused by Salmonella Enteritidis through carboxylation of osteocalcin.

BACKGROUND: The present study aimed at evaluating the effect of vitamin K (VK) supplementation on bone health of laying hens challenged by Salmonella Enteritidis. METHODS: A total of 80 32-week-old double negative salmonella-free brown-egg laying hens were randomly assigned to 4 treatments with 20 replicates each (1 bird per replicate) according to a 2 × 2 factorial design with 2 dietary VK supplementation levels [0 mg/kg (VK0) vs 2 mg/kg VK (VK2) and 2 challenge treatments [Salmonella Enteritidis (SE) vs physiological saline solution (PS)]. During the last 3 days of week 43 of age, birds of both VK treatments were either orally challenged with 1.0 mL suspension of 109 cfu/mL S. Enteritidis daily or received the same volume of PS. RESULTS: The laying rate, daily egg mass, tibia strength, CT, cOC and cOC/(cOC + ucOC) of VK2 treatment increased (P < 0.05) in contrast to VK0, however, the medullary area and ucOC of VK2 treatment decreased (P < 0.05) in contrast to VK0. Mortality, medullary area, serum Ca content of SE treatments increased (P < 0.05) in contrast to PS treatments. In both SE treatments, the decrease (P < 0.05) in birds' tibia strength was associated with higher (P < 0.05) Ca levels in serum. There is an interaction (P < 0.05) between SE challenge and VK levels with regard to tibia strength and serum Ca levels. At week 42, serum CT was positively correlated with cOC (R = 0.99, P = 0.009); at week 44, tibia strength was positively correlated with BMD (R = 0.95, P = 0.045), but negatively correlated with medullary area (R = - 0.98, P = 0.018). CONCLUSIONS: VK (2 mg/kg) supplementation to diets of laying hens can enhance bone strength under challenge situations with Salmonella Enteritidis. Medullary area has proven to be a sensitive biomarker for bone calcium loss caused by SE infection.

Novel strategies for degradation of aflatoxins in food and feed: A review.

Aflatoxins are toxic secondary metabolites mainly produced by Aspergillus fungi, posing high carcinogenic potency in humans and animals. Dietary exposure to aflatoxins is a global problem in both developed and developing countries especially where there is poor regulation of their levels in food and feed. Thus, academics have been striving over the decades to develop effective strategies for degrading aflatoxins in food and feed. These strategies are technologically diverse and based on physical, chemical, or biological principles. This review summarizes the recent progress on novel aflatoxin degradation strategies including irradiation, cold plasma, ozone, electrolyzed oxidizing water, organic acids, natural plant extracts, microorganisms and enzymes. A clear understanding of the detoxification efficiency, mechanism of action, degradation products, application potential and current limitations of these methods is presented. In addition, the development and future perspective of nanozymes in aflatoxins degradation are introduced.

Effects of sporoderm-broken spores of Ganoderma lucidum on growth performance, antioxidant function and immune response of broilers.

This study was conducted to evaluate the effects of sporoderm-broken spores of Ganoderma lucidum (SSGL), a traditional Chinese medicinal herb, on growth performance, antioxidant ability, and immunity of broilers. Three hundred male broilers with similar body weights (40.0 ± 1.0 g) at 1 d of age were assigned randomly to 4 treatments. Each treatment contained 5 replicates of 15 birds per replicate. The dietary treatments were corn-soybean meal basal diet supplemented with SSGL at the concentrations of 0 (control), 100, 200 and 500 mg/kg diet. The results showed that diets supplemented with SSGL significantly increased (P < 0.05) the average daily gain and decreased (P < 0.05) the feed:gain (F:G) ratio of birds during the finisher period (22 to 44 d of age). Moreover, the total antioxidant capability, glutathione reductase and catalase activities in the liver and spleen were significantly higher (P < 0.05) in broilers fed diets with SSGL than in broilers fed the control diet. Additionally, dietary SSGL also increased (P < 0.05) the serum interleukin (IL)-2, immunoglobulin (Ig) A and IgG levels of broilers compared with the control diet. These results suggest that SSGL have ameliorative effects on growth performance, free radical-scavenging activity, antioxidant capability, and immune function of broilers.

Efficacy of Bacillus subtilis ANSB060 Biodegradation Product for the Reduction of the Milk Aflatoxin M1 Content of Dairy Cows Exposed to Aflatoxin B1.

This study was conducted to determine the effect of Bacillus subtilis ANSB060 biodegradation product (BDP) in reducing the milk aflatoxin M1 (AFM1) content of dairy cows fed a diet contaminated with aflatoxin B1 (AFB1). Twenty-four Chinese Holstein cows (254 ± 19 d in milk; milk production 19.0 ± 1.2 kg d-1) were assigned to three dietary treatments, as follows: (1) control diet (CON), consisting of a basal total mixed ration (TMR); (2) aflatoxin diet (AF), containing CON plus 63 µg of AFB1 kg-1 of diet dry matter; and (3) aflatoxin diet plus BDP (AF + BDP), containing AF plus BDP at 0.2% of diet dry matter. The experiment lasted 12 days, including an AFB1-dosing period from days one to eight, followed by a clearance period from days nine to twelve. Milk samples were collected on days 2, 4, 6, and 8⁻12, and the plasma was sampled on day 9, before morning feeding. Short-term AFB1 exposure did not affect the milk production and composition. The plasma biochemical indices, except for lactic dehydrogenase (LDH), were also not changed by the AFB1 intake. The plasma LDH level was significantly elevated (p < 0.05) following dietary treatment with AFB1, while no significant difference was observed between the AF + BDP and CON treatments. Adding BDP to the AFB1-contaminaed diet resulted in a significant reduction in AFM1 concentration (483 vs. 665 ng L-1) in the milk, AFM1 excretion (9.14 vs. 12.71 µg d-1), and transfer rate of dietary AFB1 to milk AFM1 (0.76 vs. 1.06%). In conclusion, the addition of BDP could be an alternative method for reducing the dietary AFB1 bioavailability in dairy cows.

Dietary vitamin D3 supplementation protects laying hens against lipopolysaccharide-induced immunological stress.

BACKGROUND: The effects of vitamin D on the immune function of laying hens are not well understood. This study investigated the effects of vitamin D3 (VD3) on laying performance and immunological functions in laying hens under Escherichia coli lipopolysaccharide (LPS) challenge. METHODS: In experiment one, 360 Jinghong-1 strain layers (32 weeks) were randomly divided into four groups with six replicates per group and 15 hens per replicate. Hens were fed a basal diet supplemented with different levels of VD3 (0; 500; 1500; or 3000 IU VD3/kg of diet) for 10 weeks to determine laying performance, egg quality, and other parameters. In experiment two, 24 Jinghong laying hens (32 weeks) were fed basal diets with either 0 or 3000 IU VD3/kg of diet. After 10 weeks of feeding, six hens from each treatment were injected intravenously with 8 mg/kg of body weight of either LPS or saline. Blood and spleen samples were obtained for immune parameter analysis 4 h after injection. RESULTS: VD3 deficiency reduced egg production and egg quality; in addition, feed intake and feed-to-egg ratio increased. No significant differences were observed in these parameters except eggshell strength between dietary VD3 supplemental levels at 500; 1500; and 3000 IU VD3/kg of diet. VD3 deficiency increased serum hormone (calcitonin, parathyroid hormone, estradiol, and progesterone) and cytokine (IL-6, IL-10) levels, the ratio of IFN-γ to IL-4, myeloperoxidase activity and total IgG content in the serum, and upregulated the blood CD3+ T cell population. Splenic retinoid X receptor (RXR), nuclear factor-κB (NF-κB), inducible nitric oxide synthase (iNOS), and polymeric immunoglobulin receptor (pIgR) gene mRNA levels were upregulated in VD3-deficienct hens. VD3 deficiency significantly reduced serum Follicle stimulating hormone (FSH) and Luteinizing hormone (LH) concentrations and the number of CD4+CD25+ T cells in the blood. These changes were completely normalized by VD3 sufficiency. LPS reduced serum LH concentration, splenic lysozyme, and pIgR gene mRNA levels. LPS induced an increase in total serum IgM levels and the percentage of CD8+ T cells in the blood. The changes were completely reversed by VD3 addition. CONCLUSION: VD3 supplementation could protect laying hens not only from VD3 deficiency but also from immunological stress.

Dietary Supplementation with Chromium Picolinate Influences Serum Glucose and Immune Response of Brown-Egg Laying Hens.

To investigate the effect of chromium picolinate (CrPic) on egg production performance, egg quality, blood indices, and immune function of laying hens, 270 23-week-old Beijing Red brown-egg laying hens were randomly assigned to one of three treatments with six replicates of 15 birds per replicate for each treatment in a completely randomized design. Laying hens were fed a Cr-unsupplemented corn-soybean meal basal diet (control, containing 0.45 mg Cr/kg by analysis) or the basal diet supplemented with 0.4 or 0.6 mg Cr/kg from CrPic for 10 weeks. Laying hens fed diets supplemented with CrPic had a lower (P < 0.05) serum glucose (GLU) concentration and higher (P < 0.03) serum antibody titer against Newcastle disease than those fed the control diet at 33 weeks of age. However, supplemental CrPic had no effect (P > 0.05) on egg production and egg quality of brown-egg laying hens from 24 to 33 weeks of age. The results from this study indicated that supplemental CrPic decreased serum GLU and enhanced the immune function of brown-egg laying hens.

Ameliorating Effects of Bacillus subtilis ANSB060 on Growth Performance, Antioxidant Functions, and Aflatoxin Residues in Ducks Fed Diets Contaminated with Aflatoxins.

Bacillus subtilis ANSB060 isolated from fish gut is very effective in detoxifying aflatoxins in feed and feed ingredients. The purpose of this research was to investigate the effects of B. subtilis ANSB060 on growth performance, body antioxidant functions, and aflatoxin residues in ducks fed moldy maize naturally contaminated with aflatoxins. A total of 1500 18-d-old male Cherry Valley ducks with similar body weight were randomly assigned to five treatments with six replicates of 50 ducks per repeat. The experiment design consisted of five dietary treatments labeled as C0 (basal diet containing 60% normal maize), M0 (basal diet containing 60% moldy maize contaminated with aflatoxins substituted for normal maize), M500, M1000, and M2000 (M0 +500, 1000 or 2000 g/t aflatoxin biodegradation preparation mainly consisted of B. subtilis ANSB060). The results showed that ducks fed 22.44 ± 2.46 µg/kg of AFB1 (M0) exhibited a decreasing tendency in average daily gain (ADG) and total superoxide dismutase (T-SOD) activity in serum, and T-SOD and glutathione peroxidase (GSH-Px) activities in the liver significantly decreased along with the appearance of AFB1 and AFM1 compared with those in Group C0. The supplementation of B. subtilis ANSB060 into aflatoxin-contaminated diets increased the ADG of ducks (p > 0.05), significantly improved antioxidant enzyme activities, and reduced aflatoxin accumulation in duck liver. In conclusion, Bacillus subtilis ANSB060 in diets showed an ameliorating effect to duck aflatoxicosis and may be a promising feed additive.

Protective Effects of Sporoderm-Broken Spores of Ganderma lucidum on Growth Performance, Antioxidant Capacity and Immune Function of Broiler Chickens Exposed to Low Level of Aflatoxin B1.

This study was conducted to investigate the toxic effects of aflatoxin B1 (AFB1) and evaluate the effects of sporoderm-broken spores of Ganoderma lucidum (SSGL) in relieving aflatoxicosis in broilers. A total of 300 one-day-old male Arbor Acre broiler chickens were randomly divided into four dietary treatments; the treatment diets were: Control (a basal diet containing normal peanut meal); AFB1 (the basal diet containing AFB1-contaminated peanut meal); SSGL (basal diet with 200 mg/kg of SSGL); AFB1+SSGL (supplementation of 200 mg/kg of SSGL in AFB1 diet). The contents of AFB1 in AFB1 and AFB1+SSGL diets were 25.0 µg/kg in the starter period and 22.5 µg/kg in the finisher period. The results showed that diet contaminated with a low level of AFB1 significantly decreased (p < 0.05) the average daily feed intake and average daily gain during the entire experiment and reduced (p < 0.05) serum contents of total protein IgA and IgG. Furthermore, a dietary low level of AFB1 not only increased (p < 0.05) levels of hydrogen peroxide and lipid peroxidation, but also decreased (p < 0.05) total antioxidant capability, catalase, glutathione peroxidase, and hydroxyl radical scavenger activity in the liver and spleen of broilers. Moreover, the addition of SSGL to AFB1-contaminated diet counteracted these negative effects, indicating that SSGL has a protective effect against aflatoxicosis.

Molecular Mechanisms of Lipoic Acid Protection against Aflatoxin B1-Induced Liver Oxidative Damage and Inflammatory Responses in Broilers.

Alpha-lipoic acid (α-LA) was evaluated in this study for its molecular mechanisms against liver oxidative damage and inflammatory responses induced by aflatoxin B1 (AFB1). Birds were randomly allocated into four groups with different diets for three weeks: a basal diet, a 300 mg/kg α-LA supplementation in a basal diet, a diet containing 74 µg/kg AFB1, and 300 mg/kg α-LA supplementation in a diet containing 74 µg/kg AFB1. In the AFB1 group, the expression of GSH-PX mRNA was down-regulated (p < 0.05), and the levels of lipid peroxide and nitric oxide were increased (p < 0.05) in the chicken livers compared to those of the control group. Additionally, the mRNA level of the pro-inflammatory factor interleukin-6 was up-regulated significantly (p < 0.05), the protein expressions of both the nuclear factor kappa B (NF-κB) p65 and the inducible nitric oxide synthase were enhanced significantly (p < 0.05) in the AFB1 group. All of these negative effects were inhibited by α-LA. These results indicate that α-LA may be effective in preventing hepatic oxidative stress, down-regulating the expression of hepatic pro-inflammatory cytokines, as well as inhibiting NF-κB expression.

Protective Effects of Bacillus subtilis ANSB060 on Serum Biochemistry, Histopathological Changes and Antioxidant Enzyme Activities of Broilers Fed Moldy Peanut Meal Naturally Contaminated with Aflatoxins.

The aim of this study was to investigate the toxic effects of aflatoxins and evaluate the effectiveness of Bacillus subtilis ANSB060 in detoxifying aflatoxicosis in broilers. A total of 360 one-week-old male broilers (Ross 308) were assigned to six dietary treatments for five weeks. The treatment diets were: C0 (basal diet); C1.0 (C0 + 1.0 g B. subtilis ANSB060/kg diet); M0 (basal diet formulated with moldy peanut meal); M0.5, M1.0 and M2.0 (M0 + 0.5, 1.0 and 2.0 g B. subtilis ANSB060/kg diet, respectively). The contents of aflatoxin B1, B2, G1 and G2 in the diets formulated with moldy peanut meal were 70.7 ± 1.3, 11.0 ± 1.5, 6.5 ± 0.8 and 2.0 ± 0.3 µg/kg, respectively. The results showed that aflatoxins increased (p < 0.05) serum aspartate transaminase activity, decreased (p < 0.05) serum glutathione peroxidase activity, and enhanced (p < 0.05) malondialdehyde contents in both the serum and liver. Aflatoxins also caused gross and histological changes in liver tissues, such as bile duct epithelium hyperplasia, vacuolar degeneration and lymphocyte infiltration. The supplementation of ANSB060 reduced aflatoxin levels in the duodenum and counteracted the negative effects of aflatoxins, leading to the conclusion that ANSB060 has a protective effect against aflatoxicosis and this protection is dose-related.