Effects of dietary addition of heat-killed Mycobacterium phlei on growth performance, immune status and anti-oxidative capacity in early weaned piglets.

The contradiction between high susceptibility of early weaned piglets to enteric pathogens and rigid restriction of antibiotic use in the diet is still prominent in the livestock production industry. To address this issue, the study was designed to replace dietary antibiotics partly or completely by an immunostimulant, namely heat-killed Mycobacterium phlei (M. phlei). Piglets (n = 192) were randomly assigned to one of the four groups: (1) basal diet (Group A), (2) basal diet + a mixture of antibiotics (80 mg/kg diet, Group B), (3) basal diet + a mixture of antibiotics (same as in Group B, but 40 mg/kg diet) + heat-killed M. phlei (1.5 g/kg diet) (Group C) and (4) basal diet + heat-killed M. phlei (3 g/kg diet) (Group D). All piglets received the respective diets from days 21 to 51 of age and were weaned at the age of 28 d. Compared with the Control (Group A), in all other groups the average daily gain, average daily feed intake, small intestinal villus height:crypt depth ratio and protein levels of occludin and ZO-1 in the jejunal mucosa were increased. A decreased incidence of diarrhoea in conjunction with an increased sIgA concentration in the intestinal mucosa and serum IL-12 and IFN-γ concentrations was found in groups supplemented with heat-killed M. phlei (Groups C and D), but not in Group B. Groups C and D also showed decreased IL-2 concentrations in the intestinal mucosa with lower TLR4 and phosphor-IκB protein levels. The antioxidant capacity was reinforced in Groups C and D, as evidenced by the reduction in malondialdehyde and enhanced activities of antioxidant enzymes in serum. These data indicate that heat-killed M. phlei is a promising alternative to antibiotic use for early weaned piglets via induction of protective immune responses.

Corrigendum: Intestinal Morphologic and Microbiota Responses to Dietary Bacillus spp. in a Broiler Chicken Model.

[This corrects the article DOI: 10.3389/fphys.2018.01968.].

Effects of an immune stimulant, inactivated Mycobacterium phlei, on the growth performance as well as meat quality of fattening pigs.

Inactivated mycobacterium phlei (M. phlei) is well known for its immune-stimulatory functions in humans and livestock, but less information is available about the influence on meat quality of pigs when used as a feed additive. This study was designed to evaluate the effects of inactivated M. phlei on growth performance as well as meat quality of fattening pigs. A total of 240 cross-bred pigs ([Landrace × Yorkshire] × Duroc) with initial body weight of 80.14 ± 0.29 kg were randomly allocated to five treatments, each of which consisted of eight replicates with 6six pigs per replicate. The basal diet supplemented with five levels of inactivated M. phlei preparations (0, 3.5 × 109 [0.1% w/w], 7 × 109 [0.2%], 1.4 × 1010 [0.4%] or 2.1 × 1010 [0.6%] colony-forming units/kg) was respectively fed to the control group and four treatment groups for 30 days. Adding 0.4% of inactivated M. phlei to diet increased the average daily feed intake and average daily gain of pigs. Importantly, intramuscular fat percentage in the Longissimus dorsi (LD) was increased by feeding diet containing 0.2%, 0.4% and 0.6% of inactivated M. phlei, despite the pH value, drip loss, cooking loss and filter paper fluid uptake not being influenced. Analysis of the fatty acid components showed that some saturated fatty acids were decreased in LD after feeding inactivated M. phlei, but some monounsaturated fat acids (MUFAs) and polyunsaturated fatty acids were increased (PUFAs), which induced the total contents of MUFAs and PUFAs were improved. RT-PCR assay revealed that feeding inactivated M. phlei up-regulated genes implicated in fat metabolism in muscle, including ELOVL6, FASN, SCD1 and H-FABP. This study revealed that feeding inactivated M. phlei not only increased growth performance of fattening pigs, but also improved the meat quality by increasing intramuscular fat content, thus inactivated M. phlei probably has high utilization value in modern pig production.

Intestinal Morphologic and Microbiota Responses to Dietary Bacillus spp. in a Broiler Chicken Model.

Dietary inclusion of probiotic Bacillus spp. beneficially affect the broiler chickens by balancing the properties of the indigenous microbiota causing better growth performance. The effects of three Bacillus spp. on the growth performance, intestinal morphology and the compositions of jejunal microflora were investigated in broiler chickens. A total of 480 1-day-old male Arbor Acres broilers were randomly divided into four groups. All groups had six replicates and 20 birds were included in each replicate. The control birds were fed with a corn-soybean basal diet, while three treatment diets were supplemented with Bacillus coagulans TBC169, B. subtilis PB6, and B. subtilis DSM32315 with a dosage of 1 × 109 cfu/kg, respectively. The experiment lasted for 42 days. The compositions and diversity of jejunal microflora were analyzed by MiSeq high-throughput sequencing. The B. coagulans TBC169 group showed marked improvements of growth performance, nutrient digestibility and intestinal morphology compared with the other B. subtilis treatments. B. coagulans TBC169 supplementation improved the average body weight (BW), average daily weight gain (ADG), total tract apparent digestibility of crude protein and gross energy (GE), and reduced feed conversion rate (FCR) compared with the control group (P < 0.05). The villus height to crypt depth ratio (VH/CD) of jejunum and duodenum was increased in the birds fed with B. coagulans TBC169 compared with the control group (P < 0.05). However, two B. subtilis treatments presented more positive variation of the jejunum microflora of chickens than that in the B. coagulans TBC169 group. B. subtilis PB6 and B. subtilis DSM32315 treatments improved the diversity of jejunal microbiota on day 21 compared with the control (P < 0.05), while which were decreased on day 42 (P < 0.05). The supplementation with B. coagulans TBC169 significantly improved the proportion of Firmicutes, otherwise two B. subtilis significantly improved the proportion of Proteobacteria, Bacteroidetes, Actinobacteria, and Acidobacteria at the phylum level during starter phase and decreased the proportion of Bacteroidetes during growing phase compared with the control. The supplementation with B.subtilis DSM32315 significantly improved the proportion of Clostridiales during starter phase, whereas two B. subtilis significantly improved the proportion of Pseudomonas, Burkholderia, Prevotella, DA101 during growing phase at the genus level compared with the control. In conclusion, the dietary supplementation with probiotic Bacillus spp. strains improved body weight and intestinal morphology in broiler chickens, which might be associated with the gut microbiota.

Effects of Armillariella tabescens mycelia on the growth performance and intestinal immune response and microflora of early-weaned pigs.

This study was performed to evaluate effects of Armillariella tabescens (A. tabescens) on the growth performance and intestinal immune response and microflora in early-weaned pigs when used as feed additive. A. tabescens mycelia were added to basal diets at concentrations of 0%, 0.1%, 0.3% or 0.9% (w/w). A total of 144 commercial cross-bred piglets were randomly allocated to one of these four diets and fed for 30 days. The growth performance of early-weaned piglets displayed improvement with diets containing 0.1% and 0.3% dried mycelia powder from A. tabescens. Supplementing with 0.1% or 0.3% A. tabescens mycelia induced a 2.6- and three-fold increase in secretory immunoglobulin A (sIgA) content in the jejunal mucosa, respectively, but had only a marginal effect on sIgA in the ileal mucosa. Expression of interleukin-2, interferon-γ, and tumor necrosis factor-α in the jejunal mucosa were elevated with A. tabescens mycelia administration. Increased amounts of Lactobacillus spp. and Bifidobacterium spp. in the jejunum, and decreased amounts of Escherichia coli in the jejunum and ileum were observed with the administration of A. tabescens-containing diets. This study demonstrated that A. tabescens had beneficial effects on the growth performance and intestinal microflora of early-weaned pigs.