Positive effects of a Clostridium butyricum-based compound probiotic on growth performance, immune responses, intestinal morphology, hypothalamic neurotransmitters, and colonic microbiota in weaned piglets.

Weaning stress in piglets can lead to poor health outcomes and reduced production. We investigated the effects of probiotics, one potential antibiotic alternative, on the growth performance, serum biochemical parameters, intestinal morphology, mucosal immunity, hypothalamic neurotransmitters, and colonic microflora in weaned piglets. Thirty-six weaned piglets were fed a basal diet, a diet supplemented with colistin sulphate antibiotic, or a diet supplemented with probiotics including Clostridium butyricum, Bacillus subtilis, and B. licheniformis. Probiotics significantly increased the feed : gain ratio, improved the average day gain from day 1 to day 28, and decreased the diarrhoea index. Probiotics also lowered the serum concentrations of AST, ALT, and ALP on day 14 and lowered the serum concentration of ALT on day 28 compared with the control. Probiotic supplementation caused fewer ileal apoptotic cells. The serum and ileal concentrations of TNF-α and IL-1ß on day 28 were significantly lowered, and the serum concentrations of IL-6 were significantly lowered on days 14 and 28. Probiotic-fed piglets exhibited higher contents of hypothalamic serotonin and dopamine as well as serum γ-aminobutyric acid along with higher colonic concentrations of butyrate and valerate on day 28. High-throughput sequencing showed 972 core operational taxonomic units among all groups, of which 48 were unique to the probiotic-treated group. The relative abundance of genus Bacillus and species Bacillus velezensis was enriched in probiotic piglets; the phylogenetic investigation of communities by the reconstruction of unobserved states indicated that amino acid metabolism, DNA repair, replication and recombination proteins, and secretion systems were enriched with probiotics. In conclusion, the Clostridium butyricum-based probiotics improved growth performance, enhanced intestinal morphology, changed hypothalamic neurotransmitters and modulated colonic microflora in weaned piglets.

Effects of different stocking densities on growth performance, antioxidant ability, and immunity of finishing broilers.

The study was conducted to evaluate the effects of different stocking densities on growth performance, antioxidant capacity, and immunity of broilers. One thousand four hundred and forty 22-day-old Lingnan Yellow broilers were randomly allotted to five different stocking density groups (8, 10, 12, 14, and 16 birds per m2 ). Each group consisted of three replicates. The results showed that 8 and 10 birds/m2 groups had higher average daily feed intake (ADFI) than the others (p < 0.05). Heat shock protein 70 (HSP70 ) in the group of 16 birds/m2 had the highest levels (p < 0.05). The group of 16 birds/m2 had the lowest total antioxidant capability (T-AOC) and total superoxide dismutase (T-SOD) activities in blood serum (p < 0.05), and significantly increased interleukin-1ß (IL-1ß) and interleukin-4 (IL-4) concentration compared to the groups of 8 and 10 birds/m2 (p < 0.05). In liver, the group of eight birds/m2 had higher T-AOC levels than that of 12, 14, and 16 birds/m2 (p < 0.05) and also higher catalase (CAT) activities than that of 14 and 16 birds/m2 (p < 0.05); the group of 10 birds/m2 had the highest T-SOD activities among all groups (p < 0.05). In conclusion, the above results suggest that stocking density of broilers up to 8 or 10 birds/m2 can prevent the negative effects on growth performance and welfare parameters in broilers.

Effects of Maternal Zinc Glycine on Mortality, Zinc Concentration, and Antioxidant Status in a Developing Embryo and 1-Day-Old Chick.

This study was conducted to investigate the effects of maternal zinc glycine (Zn-Gly) supplementation as an alternative for zinc sulfate (ZnSO4) on mortality, zinc (Zn) concentration, and antioxidant status in a developing embryo and 1-day-old chick. Six hundred 39-week-old broiler breeders were randomly assigned to 6 treatments, each treatment including 5 replicates with 20 birds each. Six treatments received a basal diet (control, 24 mg Zn/kg diet) or a basal diet supplemented with ZnSO4 (80 mg Zn/kg) or Zn-Gly (20, 40, 60, or 80 mg Zn/kg), respectively. The experiment lasted for 8 weeks after a 4-week pre-experiment with a basal diet. At the last week, 100 eggs per replicate were randomly collected for incubation. Compared with the control treatment, Zn supplementation decreased (P < 0.05) embryo mortalities of the late stage and the whole period, increased (P < 0.05) liver Zn concentration in the embryo of d9, d19, and 1-day-old chick, and improved (P < 0.05) antioxidant status in the embryo of d19 and 1-day-old chick. Compared with the ZnSO4 treatment, 80 mg Zn/kg Zn-Gly treatment significantly decreased (P < 0.05) the late stage embryo mortality and increased (P < 0.05) liver Zn concentration in the embryo of d9, d19, and 1-day-old chick. The 80 mg Zn/kg Zn-Gly treatment significantly increased (P < 0.05) copper-zinc superoxide dismutase activity in d19 embryo and 1-day-old chick, total superoxide dismutase activity in 1-day-old chick, and copper-zinc superoxide dismutase messenger RNA (mRNA) abundance of d9 embryo and 1-day-old chick than that in ZnSO4 treatment. The liver metallothionein concentration of the developing embryo and 1-day-old chick and its mRNA abundance of d19 embryo were also significantly increased (P < 0.05) in the 80 mg Zn/kg Zn-Gly treatment in comparison with ZnSO4 treatment. In conclusion, maternal Zn supplementation decreased embryo mortalities of the late stage and the whole period by increasing liver Zn concentration and antioxidant status in d19 embryo and 1-day-old chick, and 80 mg Zn/kg from Zn-Gly treatment was the optimum choice.

Effects of Zinc Glycinate on Productive and Reproductive Performance, Zinc Concentration and Antioxidant Status in Broiler Breeders.

An experiment was conducted to investigate the effects of zinc glycinate (Zn-Gly) supplementation as an alternative for zinc sulphate (ZnSO4) on productive and reproductive performance, zinc (Zn) concentration and antioxidant status in broiler breeders. Six hundred 39-week-old Lingnan Yellow broiler breeders were randomly assigned to 6 groups consisting of 4 replicates with 25 birds each. Breeders were fed a basal diet (control group, 24 mg Zn/kg diet), basal diet supplemented with 80 mg Zn/kg diet from ZnSO4 or basal diet supplemented with 20, 40, 60 and 80 mg Zn/kg diet from Zn-Gly. The experiment lasted for 8 weeks after a 4-week pre-test with the basal diet, respectively. Results showed that Zn supplementation, regardless of sources, improved (P < 0.05) the feed conversion ratio (kilogram of feed/kilogram of egg) and decreased broken egg rate, and elevated (P < 0.05) the qualified chick rate. Compared with the ZnSO4 group, the 80 mg Zn/kg Zn-Gly group significantly increased (P < 0.05) average egg weight, fertility, hatchability and qualified chick rate, whereas it decreased (P < 0.05) broken egg rate. The Zn concentrations in liver and muscle were significantly higher (P < 0.05) in 80 mg Zn/kg Zn-Gly group than that in ZnSO4 group. Compared with ZnSO4 group, 80 mg Zn/kg Zn-Gly group significantly elevated (P < 0.05) the mRNA abundances of metallothionein (MT) and copper-zinc superoxide (Cu-Zn SOD), as well as the Cu-Zn SOD activity and MT concentration in liver. Moreover, the 80 mg Zn/kg Zn-Gly group had higher (P < 0.05) serum T-SOD and Cu-Zn SOD activities than that in the ZnSO4 group. This study indicated that supplementation of Zn in basal diet improved productive and reproductive performance, Zn concentration and antioxidant status in broiler breeders, and the 80 mg Zn/kg from Zn-Gly was the optimum choice for broiler breeders compared with other levels of Zn from Zn-Gly and 80 mg/kg Zn from ZnSO4.

Effects of dietary supplementation of probiotic, Clostridium butyricum, on growth performance, immune response, intestinal barrier function, and digestive enzyme activity in broiler chickens challenged with Escherichia coli K88.

BACKGROUND: Colibacillosis caused by enterotoxigenic Escherichia coli (E. coli) results in economic losses in the poultry industry. Antibiotics are usually used to control colibacillosis, however, E. coli has varying degrees of resistance to different antibiotics. Therefore the use of probiotics is becoming accepted as an alternative to antibiotics. In this study, we evaluated the effects of Clostridium butyricum (C. butyricum) on growth performance, immune response, intestinal barrier function, and digestive enzyme activity in broiler chickens challenged with Escherichia coli (E. coli) K88. METHODS: The chickens were randomly divided into four treatment groups for 28 days. Negative control treatment (NC) consisted of birds fed a basal diet without E. coli K88 challenge and positive control treatment (PC) consisted of birds fed a basal diet and challenged with E. coli K88. C. butyricum probiotic treatment (CB) consisted of birds fed a diet containing 2 × 10(7) cfu C. butyricum/kg of diet and challenged with E. coli K88. Colistin sulfate antibiotic treatment (CS) consisted of birds fed a diet containing 20 mg colistin sulfate/kg of diet and challenged with E. coli K88. RESULTS: The body weight (BW) and average day gain (ADG) in the broilers of CB group were higher (P < 0.05) than the broilers in the PC group overall except the ADG in the 14-21 d post-challenge. The birds in CB treatment had higher (P < 0.05) concentration of tumor necrosis factor-α (TNF-α) at 3 and 7 d post-challenge, and higher (P < 0.05) concentration of interleukin-4 (IL-4) at 14 d post-challenge than those in the PC treatment group. The concentration of serum endotoxin in CB birds was lower (P < 0.05) at 21 d post-challenge, and the concentrations of serum diamine oxidase in CB birds were lower (P < 0.05) at 14 and 21 d post-challenge than in PC birds. Birds in CB treatment group had higher (P < 0.05) jejunum villi height than those in PC, NC, or CS treatment at 7, 14, and 21 d post-challenge. In comparison to PC birds, the CB birds had lower (P < 0.05) jejunum crypt depth during the whole experiment. The birds in CB or CS treatment group had higher (P < 0.05) activities of amylase and protease at 3, 7, and 14 d post-challenge, and higher (P < 0.05) activity of lipase at 3, 7 d post-challenge than PC birds. CONCLUSIONS: In all, these results indicate that dietary supplementation with C. butyricum promotes immune response, improves intestinal barrier function, and digestive enzyme activities in broiler chickens challenged with E. coli K88. There is no significant difference between the C. butyricum probiotic treatment and the colistin sulfate antibiotic treatment. Therefore, the C. butyricum probiotic may be an alternative to antibiotic for broiler chickens.

The Protective Effects of Different Sources of Maternal Selenium on Oxidative Stressed Chick Embryo Liver.

The experiment was conducted to investigate the protective effects of different sources of maternal selenium (Se) on oxidative stressed chick embryo. A total of 270 Lingnan Yellow broiler breeders were randomly allocated into three treatments with five replicates for 18 birds each. Breeders were fed with basal diet (BD) including 0.04 mg/kg Se or BD supplemented with sodium selenite (SS) or selenomethionine (SM) at a level of 0.15 mg Se/kg. The rearing experiment lasted for 8 weeks after an 8-week pre-test. Twenty eggs were collected from each replicate during the last 10-day, then incubated in a commercial incubator. On embryonic 17th, fertile eggs were transferred into 39.5 °C temperature stimulation for 6 h. Afterward, five eggs were randomly selected from each replicate for collecting chick embryo sample. The results showed that Se supplementation in the diet of breeders resulted in lower reactive oxygen species (ROS), heat shock protein 70 (HSP70), malondialdehyde (MDA), carbonyl and 8-hydroxydeoxyguanosine (8-OHdG) concentrations and higher glutathione peroxidase (GPx), total superoxide dismutase (T-SOD), and catalase (CAT) activities in heat stress treated chick embryo (P < 0.05), and ROS, MDA, carbonyl, 8-OHdG concentrations in SM treatment were lower than those in SS treatment (P < 0.05). Se supplementation elevated cellular glutathione peroxidase (GPx1) mRNA level and activity, cytoplasmic thioredoxin reductase (TrxR1) activity and selenoprotein P (SelP) mRNA and protein level (P < 0.05), and maternal SM showed a higher value than maternal SS in upregulating GPx1, TrxR1, and SelP mRNA levels as well as GPx1 and TrxR1 activities or SelP protein level (P < 0.05). This study indicated that maternal Se can enhance antioxidative capacity and reduce ROS concentration and oxidative damage by upregulating the expression of antioxidative selenoprotein, and maternal SM is superior to SS in heat stress treated chick embryo.

Effects of fluoride on hepatic antioxidant system and transcription of Cu/Zn SOD gene in young pigs.

Thirty-two barrows (Duroc x Landrace x Yorkshire) were randomly divided into four groups, each of which included eight pigs. The groups received the same basal diet supplemented with 0, 100, 250 and 400mg/kg fluoride, respectively. The malondialdehyde (MDA) and glutathione (GSH) levels, antioxidant enzymes activities and zinc/copper superoxide dismutase (Cu/Zn SOD) mRNA content in the liver were determined to evaluate the fluoride hepatic intoxication. Results showed the increased lipid peroxides (LPO) level and the reduced GSH content, along with a concomitant decrease in the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px). Moreover, the level of hepatic Cu/Zn SOD mRNA was also significantly reduced. We suggest the mechanism of fluoride injuring the liver as follows: fluoride causes a decrease in Cu/Zn SOD mRNA and the reduced activities of antioxidant enzymes, leads to the declined ability of scavenging free radicals with excessive production of LPO, which seriously damages the hepatic structure and function.

Evaluation of caspase-dependent apoptosis during fluoride-induced liver lesion in pigs.

Sixteen barrows (Duroc x Landrace x Yorkshire) were randomly divided into two groups, each consisting eight pigs. The groups received the same basal diet supplemented with 0 and 400 mg/kg fluoride, respectively. Histological examinations, including in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), Haematoxylin and Eosin staining (HE) and transmission electron microscopy observation, found apoptotic hepatocytes 50 days after additional 400 mg/kg fluoride treatment. The obvious DNA ladder and the significantly increased both hepatic caspase-9 and caspase-3 activity indicated that fluoride induced caspase-dependent apoptosis in vivo. In addition, serum glutamate pyruvate transaminase (GPT) activity and hepatic lipid peroxides (LPO) concentration was significantly increased. The activity of serum glutamate oxaloacetate transaminase (GOT) showed an increased trend. The results suggest that fluoride induces apoptosis by elevating the oxidative stress-induced lipid peroxidation, causing mitochondrial dysfunction and further activating caspase-9 and caspase-3.