Piglet growth performance improved by dietary supplementation of porous or nano particles of zinc oxide may be related to the gut microbiota.

Previous studies on porous or nano particles zinc oxide (ZnO) in the piglets have mainly focused on growth performance and intestinal inflammation, but have scarcely explored the efficacy on gut microbiota. In addition, the efficacy of nano particles ZnO, which is related to its product quality, remains undefined. This study aimed to determine the efficacy of dietary 500 mg/kg porous or nano particles ZnO on the growth performance and gut microbiota of the weaned piglets. A total of 128 weaned piglets were randomly assigned to the dietary groups: NC (basal diet), PC (basal diet + 3,000 mg/kg conventional ZnO), 500HiZ (basal diet + 500 mg/kg porous particles ZnO), and 500ZNP (basal diet + 500 mg/kg nano particles ZnO). Compared with the NC diet group, both 500HiZ and 500ZNP increased (P < 0.05) average daily feed intake (1 to 28 d) and average daily gain (1 to 28 d), and the 500ZNP tended to decrease feed to gain ratio (F:G ratio, 1 to 28 d) (P = 0.09). Both 500HiZ and 500ZNP decreased crypt depth of the ileum and increased claudin-2 in the duodenum and zonula occludens-1 in the ileum (P < 0.05). Moreover, both 500HiZ and 500ZNP decreased IL-1ß and tumor necrosis factor-α (TNF-α) in the jejunum and decreased TNF-α and IL-6 in the ileum (P < 0.05). Both 500HiZ and 500ZNP increased microbial ß-diversity index in the ileum and microbial α-diversity indices in the colon of piglets (P < 0.05). The probiotic genera Coprococcus (500ZNP) and Blautia (500HiZ) were positively correlated with the F:G ratio (1 to 28 d) in colon of piglets (P < 0.05). In addition, 500HiZ promoted mitochondrial fusion protein 1 (MFN1) and zinc transporter-1 (ZnT-1) in the jejunum (P < 0.05), whilst 500ZNP decreased MFN1 in the jejunum and ZnT-1 in the ileum (P < 0.05). In summary, both 500HiZ and 500ZNP improved the growth performance of piglets, which is likely via the genera Blautia and Coprococcus, respectively. Both 500HiZ and 500ZNP improved barrier function and inflammation of the intestine, and 500HiZ achieved better efficacy than 500ZNP on intestine mitochondrial functions.

Effects of Zinc Lactate Supplementation on Growth Performance, Intestinal Morphology, Serum Parameters, and Hepatic Metallothionein of Chinese Yellow-Feathered Broilers.

In poultry, organic zinc compounds have higher bioavailability than inorganic zinc sources. However, as an organic zinc source, the application of zinc lactate (ZL) on Chinese yellow-feathered broilers has been rarely reported. Hence, the present study aimed to investigate the effects of ZL supplementation on growth performance, small intestinal morphology, serum biochemical parameters, immune organ index, as well as hepatic metallothionein of Chinese yellow-feathered broilers. A total of 2100 broilers (19 days old) were randomly assigned to 5 treatment groups, including the control (fed basal diet), ZL40 (basal diet plus 40 mg/kg ZL), ZL60 (basal diet plus 60 mg/kg ZL), ZL80 (basal diet plus 80 mg/kg ZL), and ZS80 (basal diet plus 80 mg/kg ZS. Each treatment group had 6 replicates with 70 chickens per replicate. Compared to the control group, the ZL40 and the ZS80 groups had a lower feed to gain ratio (P < 0.05), ZL40 group had higher duodenum and ileum villus heights (P < 0.05), and ZS80 and ZL80 groups had a lower ratio of villus height to crypt depth in the jejunum (P < 0.01). In addition, the ZL60 group had a higher concentration of total protein (P < 0.05) and activity of glutathione peroxidase (GSH-Px) (P < 0.01) compared with the ZS80 and the control groups. Interestingly, the ZL40, ZL60, and ZL80 groups all had higher levels of hepatic metallothionein than the other groups (P < 0.01). In conclusion, zinc lactate had a higher bioavailability and could be used as an alternative to zinc sulfate.

Research Note: Effects of glycerol monolaurate supplementation on egg production, biochemical indices, and gut microbiota of broiler breeders at the late stage of production.

The objective of the present study was to determine the effect of glycerol monolaurate (GML) supplementation on egg production, biochemical indices, and gut microbiota of broiler breeders at the late stage of production. Total of 180 healthy Qingyuan Partridge broiler breeders were randomly assigned to 2 groups: 1) corn-soybean meal based diet, and 2) basal diet supplemented with 300 mg glycerol monolaurate/kg. Each treatment group had 6 replicates with 15 birds within each replicate. The experiment started at wk 33 and lasted for 8 weeks. Feed conversion rate, egg weight, egg shape index, shell breaking strength, and shell thickness were not different between control and treatment groups. Supplementation of GML significantly decreased the egg breaking rate. All blood chemical indices and antioxidant parameters were not affected by GML except total antioxidant capacity which increased significantly with GML supplementation. Alpha diversity indices (Shannon, Simpson, Chao1, Ace, goods_coverage, and PD_whole tree) were not different between the 2 groups. Composition of cecal microbiota was not affected by GML supplementation except Euryarchaeota and Proteobacteria at phylum level. Overall, supplementation of glycerol monolaurate at 300 mg/kg level improved eggshell quality but its effect on cecal microbiota composition was limited on broiler breeders at the late stage of production.

Effects of dietary rosemary extract supplementation on growth performance, nutrient digestibility, antioxidant capacity, intestinal morphology, and microbiota of weaning pigs.

Rosemary (Rosmarinus officinalis L.) extract (RE) has multiple pharmacological and biological activities, including the use as a food additive and medicine. This study was conducted to investigate the effects of dietary RE supplementation on the growth performance, nutrient digestibility, antioxidant capacity, intestinal morphology, and microbiota of weaning piglets. A total of 192 crossbred weaned piglets [Duroc × (Large White × Landrace)] (initial body weight = 6.65 ± 0.33 kg, weaned days = 23 ± 1 d) were group housed (six pigs per pen; n = 8 pens/treatment). Pigs were fed a corn-soybean meal-based control diet or the basal diet supplemented with 100, 200, or 400 mg/kg RE. Pigs were allowed ad libitum access to fed for 21 d. The growth performance and apparent total tract digestibility of nutrients, and intestinal morphology and antioxidant status were evaluated. The components of the microbial microflora were also determined in the cecal samples. Compared with the control, dietary supplementation with RE increased the final body weight, average daily gain, and average daily feed intake (linear, P = 0.038, 0.016, and 0.009, respectively), and decreased the diarrhea ratio in piglets (linear, P < 0.05). The digestibility of crude protein (linear, P = 0.034) and gross energy (linear, P = 0.046) increased with treatment with RE. Piglets fed RE showed longer villus height (linear, P = 0.037 and 0.028, respectively) and villus height/crypt depth (linear, P = 0.004 and 0.012; quadratic, P = 0.023 and 0.036, respectively) in the jejunum and ileum, in addition to a lesser crypt depth in the jejunum (linear, P = 0.019) and ileum (quadratic, P = 0.042). The addition of RE increased the activity of superoxide dismutase (linear, P = 0.035 and 0.008, respectively) and glutathione peroxidase activity (linear, P = 0.027 and 0.039, respectively) and decreased the content of malondialdehyde (linear, P = 0.041 and 0.013; quadratic, P = 0.023 and 0.005, respectively) in the serum and liver. Dietary RE supplementation, compared with the control, increased the number of Bifidobacterium (linear, P = 0.034) and Bacteroidetes (linear, P = 0.029), while decreased Escherichia coli (linear, P = 0.008; quadratic, P = 0.014) in the cecal contents. Thus, dietary RE supplementation can improve growth performance, nutrient digestibility, antioxidant capacity, intestinal morphology, and the microbiota in weaned piglets, and 200 mg/kg may be considered the optimum dosage.

Effects of monobutyrin supplementation on egg production, biochemical indexes, and gut microbiota of broiler breeders.

The objective of the present study was to determine the effect of monobutyrin supplementation on egg production, biochemical indexes, and gut microbiota of broiler breeders at the late stage of production. A total of 180 healthy Qingyuan partridge broilers were randomly assigned to 2 groups: 1) corn-soybean meal-based diet and 2) basal diet supplemented with 250 mg monobutyrin/kg. Each treatment group had 6 replicates/cages with 15 birds within each replicate. The experiment started at week 33 and lasted for 8 wk. Egg production rate, feed conversion rate, shell breaking strength, and shell thickness were not different between control and treatment groups. Supplementation of monobutyrin increased egg weight and tended to decrease egg breaking rate of Qingyuan partridge chickens. Supplementation of monobutyrin did not affect any of the biochemical indexes except total protein concentration. The 4 antioxidant parameters measured were not affected either. Alpha diversity indexes (Shannon, Simpson, Chao1, Ace, and Good's Coverage) and composition of cecal microbiota were not affected by monobutyrin supplementation. Overall, supplementation of monobutyrin at 250 mg/kg level improved egg quality, but its effect on cecal microbiota composition was limited.

Transcriptional regulation of G2/M regulatory proteins and perturbation of G2/M Cell cycle transition by a traditional Chinese medicine recipe.

ETHNOPHARMACOLOGICAL RELEVANCE: Hedyotis diffusa Willd. (H) and Scutellaria barbata D.Don (S) are ancient anti-cancer Chinese herb medicines. When combined, known as HS, it is one of the most commonly prescribed Chinese Medicines for cancer patients today in China. AIM OF THE STUDY: The prevention of disease progression is a dominant concern for the growing number of men with prostate cancer. The purpose of this work is to evaluate the action and mode of action of Chinese Medicine recipe HS in inhibiting prostate cancer progression in preclinical models. METHODS: Effects of HS were analyzed in prostate cancer cell lines by evaluating proliferation, cell cycle profile, DNA damage and key regulators responsible for G2 to M phase transition. The transcriptional activities of these regulators were determined by RT-PCR and ChIP. The efficacy of HS in vitro was validated in an animal model. RESULTS: HS treatment was observed to reduce DNA content and accumulated prostate cancer cells at the G2/M phase. Immunolabeling for phospho-Histone H3 in association with nocodazole to capture mitotic cells confirmed that HS impeded G2 to M transition. After excluding DNA damage-induced G2 arrest, it was revealed that HS reduced expression of Cyclin B1, CDK1, PLK1 and Aurora A at both protein and mRNA levels, with concomitant reduction of H3K4 tri-methylation at their promoter-regions. Animals that received oral administration of HS with a dosage relevant to clinical application showed reduced tumor volume and weight with a reduction of Cyclin B1, CDK1, PLK1 and Aurora A protein levels. CONCLUSIONS: HS acts by impeding the G2 to M transition of prostate cancer cells. It is likely that the mode of action is transcriptionally suppressing proteins governing mitotic entry, without eliciting significant DNA damage.

Effects of dietary supplementation of Lycium barbarum polysaccharides on growth performance, immune status, antioxidant capacity and selected microbial populations of weaned piglets.

Lycium barbarum polysaccharides (LBPs) are a complex mixture of highly branched and partially characterised polysaccharides and proteoglycans extracted from the goji berry. This mixture has great potential as a novel feed supplement for pigs. Two trials were conducted to evaluate the effects of supplementation with LBPs on the growth performance, immune status, antioxidant capacity and selected intestinal microbial populations in weaned piglets. In trial 1, a total of 400 weaned piglets [(Yorkshire × Landrace) × Duroc] with an average body weight (BW) of 6.34 ± 0.16 kg (21 days of age) were divided into five groups and fed a basal diet (control group) or a basal diet containing 1,000, 2,000, 4,000 or 6,000 mg/kg LBPs (supplemented at the expense of corn). Supplementation with 4,000 or 6,000 mg/kg LBPs for 2 weeks significantly increased the average daily gain (ADG) and average daily feed intake (ADFI) of the pigs compared with the control group (p < .05). In trial 2, thirty-two 21-days-old weaned piglets (BW: 6.33 ± 0.11 kg) were allotted to a control group (fed with a basal diet) or an experimental group (basal diet containing 4,000 mg/kg LBPs). The experiment lasted for 14 days. Pigs fed LBP diets exhibited an increased ADG and ADFI, and a decreased diarrhoeal incidence compared with those fed the basal diets (p < .05). Supplementation with LBPs increased the serum IgG and IgM levels (p < .05). Dietary LBPs effectively promoted antioxidant defence properties through enhancing the activities of serum, liver superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), in addition to decreasing the malondialdehyde (MDA) content (p < .05). The addition of LBPs increased the amounts of Bacteroidetes in the ileum and caecum and the caecal contents of Lactobacillus spp. and Bifidobacterium spp. (p < .05), while decreased the populations of Escherichia coli and Firmicutes in the ileum and caecum (p < .05) compared with the control group. Our results suggest that dietary supplementation with LBPs can enhance growth performance, immune status and antioxidant capacity, and improve the intestinal microbial populations of weaned piglets.

The effects of dietary supplementation with porous zinc oxide on growth performance, intestinal microbiota, morphology, and permeability in weaned piglets.

The objective of this experiment was to evaluate the effects of dietary supplementation with porous zinc oxide (HiZox) on growth performance, intestinal microbiota, morphology, and permeability in weaned piglets. A total of 128 weaned piglets [(Landrace × Yorkshire) × Duroc] with an average body weight (BW) of (6.55 ± 0.25 kg; 21 d of age) were randomly assigned to four dietary treatments: (1) a corn-soybean basal diet; (2) basal diet + 3,000 mg/kg conventional ZnO; (3) basal diet + 200 mg/kg HiZox; (4) basal diet + 500 mg/kg HiZox. The experiments lasted for 28 days. Incremental HiZox in the diet increased ADG (linear p = 0.015; quadratic p = 0.043) and ADFI (linear p = 0.027; quadratic p = 0.038), and the diarrhea index decreased linearly and quadratically (p < 0.01) as HiZox supplemented increased. Furthermore, supplementation with HiZox increased the amounts of Lactobacillus spp. (p < 0.05) in the ileum and cecum in comparison with that of control treatment or 3,000 mg/kg ZnO treatment, while decreased the populations of Escherichia coli, Clostridium coccoides, and Clostridium. leptum subgroup (p < 0.05) in the ileum and cecum relative to those in control treatment. The addition of HiZox increased the villus height and villus-to-crypt ratio (VC) of duodenum, jejunum, and ileum (p < 0.05), while decreased the crypt depth of jejunum (p < 0.05) and tended to reduce the crypt depth of duodenum (p < 0.10) compared with the control treatment. Piglets fed with 500 mg/kg HiZox had lower serum D-lactate and diamine oxidase (DAO) than those fed with basal control diet or 3,000 mg/kg ZnO diet (p < 0.01). The results suggested that supplementation with HiZox modulated intestinal microbial composition and improved intestinal morphology, which may exert protective effects on the integrity of the mucosal barrier function of weaned piglets, was as efficacious as pharmaceutical doses of ZnO in enhancing growth performance, indicating that the HiZox may be a promising alternative to pharmaceutical doses of ZnO.

Dietary sulfur amino acids affect jejunal cell proliferation and functions by affecting antioxidant capacity, Wnt/ß-catenin, and the mechanistic target of rapamycin signaling pathways in weaning piglets.

Intestinal epithelial cells undergo rapid renewal along the crypt-villus axis (CVA), which ensures intestinal functions. Weaning stress differentially effects intestinal epithelial cell metabolism and physiological states along the CVA. Sulfur amino acids (SAA) play a key role in intestinal epithelial cell functioning. This study evaluated the effects of SAA dietary supplementation on weaning pig jejunal epithelial cells along the CVA. Sixteen Duroc × Landrace × Yorkshire piglets (6.16 ± 0.22 kg BW) were weaned at 21 d of age and were blocked by BW and gender and the randomly assigned to 1 of 2 groups fed diets consisting of low (0.53%) or high (0.85%) levels of SAA for a 7-d period. All piglets were euthanized for tissue sampling on day 7 postweaning. Jejunal epithelial cells were isolated along the CVA to yield 3 "cell fractions" (upper villus, middle villus, and crypt cells). The number of proliferating cells per crypt of piglets fed the high SAA diet was lower (P < 0.05) than that for low SAA diet. High SAA diet piglets tended to have decreased (P = 0.059) sucrase activities compared low SAA diet piglets. A high SAA diet increased (P < 0.05) total antioxidant capacity, catalase, and superoxide dismutase activities compared with a low SAA diet. mRNA expression levels of claudin-1, Slc5a1, and Slc7a9 in high SAA diet piglets were lower (P < 0.05) than for low SAA diet piglets. There were no interactions between dietary SAA and cell sections along the CVA for enzyme activities and mRNA expression in any of the weaned piglets. Protein amounts and phosphorylation levels related to Wnt/ß-catenin and mechanistic targeting of rapamycin (mTOR) signaling pathways were affected by SAA in weaning piglets. These findings indicate that dietary SAA affects jejunal cell proliferation and functions in weaning piglets. There appears to be no interactions between dietary SAA and cell sections along the CVA. The effects of SAA may be partly through affecting antioxidant capacity, and Wnt/ß-catenin and mTOR signaling pathway.

Correction: Comparison of porous and nano zinc oxide for replacing high-dose dietary regular zinc oxide in weaning piglets.

[This corrects the article DOI: 10.1371/journal.pone.0182550.].

Comparison of porous and nano zinc oxide for replacing high-dose dietary regular zinc oxide in weaning piglets.

The aim of this study was to compare the effect of dietary supplementation with low dose of porous and nano zinc oxide (ZnO) on weaning piglets, and to evaluate the possibility of using them as an alternative to high dose of regular ZnO. Piglets were randomly allocated into four treatment groups fed with four diets: (1) basal diet (NC), (2) NC+ 3000 mg/kg ZnO (PC), (3) NC + 500 mg/kg porous ZnO (HiZ) and (4) NC + 500 mg/kg nano ZnO (ZNP). The result showed that piglets in HiZ group had less diarrhea than ZNP group (P < 0.05). Besides, there was no significant difference between PC, HiZ and ZNP groups in terms of serum malondialdeyhde (MDA) concentration and glutathione peroxidase (GSH-Px) activity (P > 0.05). Analysis of trace metal elements revealed that piglets fed with high dose of regular ZnO had the highest Zn level in kidney (P < 0.05), which may induce kidney stone formation. Additionally, a decrease in ileal crypt depth was observed in PC, HiZ and ZNP group, suggesting an effective protection against intestinal injury. Results of mRNA analysis in intestine showed that ZNP supplementation had better effects on up-regulated trefoil factor 3 (TFF3) and nuclear factor erythroid 2-related factor 2 (Nrf2) levels in duodenum and jejunum than HiZ did (P < 0.05), implying that nano ZnO may possess higher anti-inflammatory capacity than porous ZnO. In conclusion, dietary supplementation with low dose of porous and nano ZnO had similar (even better) effect on improving growth performance and intestinal morphology, reducing diarrhea and intestinal inflammatory as high dose of regular ZnO in weaning piglets. Compared with nano ZnO, porous ZnO had better performance on reducing diarrhea but less effect on up-regulation of intestinal TFF3 and Nrf2.

Effects of yeast products on the intestinal morphology, barrier function, cytokine expression, and antioxidant system of weaned piglets.

The goal of this study was to evaluate the effects of a mixture of yeast culture, cell wall hydrolysates, and yeast extracts (collectively "yeast products," YP) on the performance, intestinal physiology, and health of weaned piglets. A total of 90 piglets weaned at 21 d of age were blocked by body weight, sex, and litter and randomly assigned to one of three treatments for a 14-d feeding experiment, including (1) a basal diet (control), (2) 1.2 g/kg of YP, and (3) 20 mg/kg of colistin sulfate (CSE). No statistically significant differences were observed in average daily feed intake, average daily weight gain, or gain-to-feed ratio among CSE, YP, and control piglets. Increased prevalence of diarrhea was observed among piglets fed the YP diet, whereas diarrhea was less prevalent among those fed CSE. Duodenal and jejunal villus height and duodenal crypt depth were greater in the control group than they were in the YP or CSE groups. Intraepithelial lymphocytes (IEL) in the duodenal and jejunal villi were enhanced by YP, whereas IEL in the ileal villi were reduced in weaned piglets fed YP. Secretion of jejunal and ileal interleukin-10 (IL-10) was higher and intestinal and serum antioxidant indexes were affected by YP and CSE. In YP- and CSE-supplemented animals, serum D-lactate concentration and diamine oxidase (DAO) activity were both increased, and intestinal mRNA expressions of occludin and ZO-1 were reduced as compared to the control animals. In conclusion, YP supplementation in the diets of weaned piglets appears to increase the incidence of diarrhea and has adverse effects on intestinal morphology and barrier function.

Effect of High Dietary Tryptophan on Intestinal Morphology and Tight Junction Protein of Weaned Pig.

Tryptophan (Trp) plays an essential role in pig behavior and growth performances. However, little is known about Trp's effects on tight junction barrier and intestinal health in weaned pigs. In the present study, twenty-four (24) weaned pigs were randomly assigned to one of the three treatments with 8 piglets/treatments. The piglets were fed different amounts of L-tryptophan (L-Trp) as follows: 0.0%, 0.15, and 0.75%, respectively, named zero Trp (ZTS), low Trp (LTS), and high Trp (HTS), respectively. No significant differences were observed in average daily gain (ADG), average daily feed intake (ADFI), and gain: feed (G/F) ratio between the groups. After 21 days of the feeding trial, results showed that dietary Trp significantly increased (P < 0.05) crypt depth and significantly decreased (P < 0.05) villus height to crypt depth ratio (VH/CD) in the jejunum of pig fed HTS. In addition, pig fed HTS had higher (P < 0.05) serum diamine oxidase (DAO) and D-lactate. Furthermore, pig fed HTS significantly decreased mRNA expression of tight junction proteins occludin and ZO-1 but not claudin-1 in the jejunum. The number of intraepithelial lymphocytes and goblet cells were not significantly different (P > 0.05) between the groups. Collectively, these data suggest that dietary Trp supplementation at a certain level (0.75%) may negatively affect the small intestinal structure in weaned pig.