Effects of sanitary conditions with lipopolysaccharide injection and dietary valine supplementation on growth performance, immune response, bacterial profile, and microbial metabolites in weaned pigs.

This study investigated the effects of dietary L-valine (Val) supplementation and sanitary conditions with lipopolysaccharide injection on growth performance, immune response, and intestinal bacterial profiles and metabolites in weaned pigs. Thirty-two weaned pigs (6.98 ± 0.47 kg) were randomly assigned to treatments in a 2 × 2 factorial arrangement based on dietary Val levels and sanitary conditions (low or high). The pigs were fed either a basal diet containing the standard levels of Val suggested by (NRC), (2012) or a basal diet supplemented with 0.1% L-Val. A room designated as a high sanitary room was washed weekly, whereas the designated low sanitary room was not washed throughout the experiment and 5 kg of manure from the nursery pig barn was spread on the pen floors on day 1. All data were analysed using a mixed procedure of SAS, with the individual pen as the experimental unit. The pigs raised in low sanitary conditions exhibited a lower (p < 0.05) average daily gain, average daily feed intake, and gain-to-feed ratio and a higher (p < 0.05) incidence of diarrhoea than those raised in high sanitary conditions during the 14-d experimental period. The pigs in the low sanitary group also had a lower (p < 0.05) concentration of butyrate in the jejunum and a higher (p < 0.05) concentration of NH3-N in the colon than those in the high sanitary group. Dietary Val supplementation was reduced (p < 0.05) plasma interleukin (IL)-1ß and IL-1 receptor antagonist concentrations as well as isovalerate and NH3-N concentrations in the colon, regardless of sanitary conditions. Interactions between dietary Val supplementation and sanitary conditions were observed in the abundances of mRNA-encoding ß-defensins 113, 125 and 129 (p < 0.05). In conclusion, dietary Val supplementation beneficially modulates inflammatory responses and microbial metabolites regardless of sanitary conditions while transcriptional levels of ß-defensins are regulated by dietary Val supplementation in a manner dependent on housing hygiene conditions.

Effects of diets with different amino acid release characteristics on the gut microbiota and barrier function of weaned pigs.

BACKGROUND: The absorption and utilization of proteins by animals is affected by the amino acid (AA) release characteristics of their diets. In the present study, we aimed to determine the effects of diets with various amino acid release characteristics on the intestinal barrier function and diversity of gut microbiota of weaned pigs. RESULTS: Forty-eight pigs (7.45 ± 0.58 kg) were fed with diets having different amino acid release characteristics during a period of 28 days. We used a 2 × 3 full-factor (two protein levels and three protein sources with differing amino acid release characteristics) experimental design, with normal (standard terminal ileal digestibility of 17.5%) or low (standard terminal ileal digestibility of 14.9%) protein levels as the first factor. Casein (CAS), corn gluten meal (CGM) and a MIX diet were used as protein sources. Due to the more balanced release of amino acids, the diamine oxidase (DAO) concentrations in the CAS and MIX groups were significantly lower than those in the CGM group (P < 0.05); Reducing the dietary protein content from 17.5% to 14.9% had no significant effects on the levels of serum DAO or D-lactic acid. By contrast, it increased the microbial diversity (chao1 and ACE values) and the number of Lactobacillus in the jejunum (P < 0.05). The CAS-containing diet and the MIX diet resulted in significantly higher microbial diversity (Simpson and Shannon) than the CGM-containing diet in the jejunum. CONCLUSION: The balanced release of amino acids in CAS and MIX diets maintained intestinal barrier function and increased gut microbiota diversity. These findings could potentially provide a scientific reference for the rational preparation of piglet feed.

Porcine ear necrosis: characterization of lesions and associated pathogens.

Porcine ear necrosis (PEN) is characterized by ulcerative lesions of the ear auricle. To investigate that problem, three farms with PEN in nursery pigs were included, and the study aim was to characterize PEN and the potential role of pathogens and mycotoxins. Within each farm, one batch of weaned piglets was included and the prevalence and severity of PEN were monitored for 6-7 weeks. Within each batch, 30 PEN-affected/non-affected animals were randomly selected. Blood samples were taken from these animals, to assess the systemic presence of pathogens and mycotoxins, as well as punch biopsies from the ear auricle for histopathological examination. From 10 animals, scrapings and swabs from the lesions were subjected to nanopore metagenomic sequencing and bacteriological cultivation, respectively. In all three farms, lesions appeared within 3-4 weeks post-weaning. The prevalence at the end of the nursery was 33%, 24%, and 46% for farms A, B, and C, respectively. Most affected pigs had mild to moderate lesions. Blood samples revealed low to very low levels of pathogens and mycotoxins. Different bacteria such as Staphylococcus, Streptococcus, Fusobacterium, Mycoplasma, and Clostridium species were identified by sequencing in the scrapings. The first two pathogens were also most often identified in bacterial cultures. Mycoplasma hyopharyngis was only found in PEN-affected pigs. Histopathological changes were primarily observed in the outer layer of the epidermis. The results suggest that PEN lesions develop by damage to the outer part of the skin e.g. by ear suckling or biting, followed by multiplication of opportunistic pathogens.

Different Fatty Acid Supplementation in Low-Protein Diets Regulate Nutrient Utilization and Lipid and Amino Acid Metabolism in Weaned Pigs Model.

This study was conducted to evaluate the effects of a low-protein (LP) diet supplemented with sodium butyrate (SB), medium-chain fatty acids (MCFAs) and n-3 polyunsaturated fatty acids (PUFAs) on nutrient utilization and lipid and amino acid metabolism in weaned pigs. A total of 120 Duroc × Landrace × Yorkshire pigs (initial body weight: 7.93 ± 0.65 kg) were randomly assigned to five dietary treatments, including the control diet (CON), LP diet, LP + 0.2% SB diet (LP + SB), LP + 0.2% MCFA diet (LP + MCFA) and LP + 0.2% n-3 PUFA diet (LP + PUFA). The results show that the LP + MCFA diet increased (p < 0.05) the digestibility of dry matter and total P in pigs compared with the CON and LP diets. In the liver of the pigs, the metabolites involved in sugar metabolism and oxidative phosphorylation significantly changed with the LP diet compared with the CON diet. Compared with the LP diet, the altered metabolites in the liver of the pigs fed with the LP + SB diet were mainly associated with sugar metabolism and pyrimidine metabolism; the altered metabolites in the liver of pigs fed with the LP + MCFA and LP + PUFA diets were mainly associated with lipid metabolism and amino acid metabolism. In addition, the LP + PUFA diet increased (p < 0.05) the concentration of glutamate dehydrogenase in the liver of pigs compared with the LP diet. Furthermore, the LP + MCFA and LP + PUFA diets increased (p < 0.05) the mRNA abundance of sterol regulatory element-binding protein 1 and acetyl-CoA carboxylase in the liver compared with the CON diet. The LP + PUFA diet increased (p < 0.05) mRNA abundances of fatty acid synthase in the liver compared with the CON and LP diets. Collectively, the LP diet supplemented with MCFAs improved nutrient digestibility, and the LP diet supplemented with MCFAs and n-3 PUFAs promoted lipid and amino acid metabolisms.

A dietary spray-dried plasma feeding programme improves growth performance and reduces faecal bacterial shedding of nursery pigs challenged with enterotoxigenic Escherichia coli K88.

This study aimed to assess the effects of different spray-dried plasma (SDP) feeding programmes to pigs on performance, intestinal histomorphology and faecal bacterial shedding after an Escherichia coli K88 challenge. A total of 96 piglets (5.77 ± 0.01 kg) were weaned at 21 days of age (Day 0) and challenged with 3 ml of 1 × 1010 CFU of E. coli K88 in total 3.0 × 1010 CFU/animal on Days 0, 2 and 4. Pigs were fed nursery diets containing 0.0%, 3.0%, 6.0% or 9.0% SDP from weaning to 35 days of age; 0.0%, 1.5%, 3.0% or 4.5% SDP from 36 to 49 days; and the same control diet (without SDP), for the last 10 days of the experiment (50-59 days of age). Performance was measured from 35 to 59 days of age and faecal bacterial shedding and intestinal histomorphometry were evaluated at Days 28 and 49 of age respectively. From 21 to 35 days of age, there was a linear effect for body weight (BW) and average daily gain (ADG), a trend of linear effect for average daily feed intake (ADFI) and a quadratic effect for feed:gain ratio (FG). From 21 to 49 days, the 9.0:4.5% and 6.0:3.0% SDP feeding programmes improved BW, ADG and FG when compared to the other treatments. At 59 days of age, BW and ADG were increased by the two highest SDP feeding programmes. The 9.0:4.5% SDP feeding programme increased ADFI from 21 to 59 days of age, with 6.0:3.0% being intermediate and the other two treatments being lowest. The CFU counts of E. coli/g of faeces decreased linearly with increasing addition of SDP. These results indicate that an extended inclusion of increased SDP levels in post-weaning diets can improve growth potential and decrease bacterial shedding induced by E. coli K88.

Agrobacterium sp. ZX09 ß-Glucan Attenuates Enterotoxigenic Escherichia coli-Induced Disruption of Intestinal Epithelium in Weaned Pigs.

To explore the protective effect of dietary ß-glucan (BGL) supplementation on intestinal epithelium exposure to enterotoxigenic Escherichia coli (ETEC), thirty-two weaned pigs were assigned to four groups. Pigs were fed with a basal diet or basal diet containing 500 mg/kg BGL, and were orally infused with ETEC or culture medium. Results showed BGL supplementation had no influence on growth performance in weaned pigs. However, BGL supplementation increased the absorption of D-xylose, and significantly decreased the serum concentrations of D-lactate and diamine oxidase (DAO) in the ETEC-challenged pigs (p < 0.05). Interestingly, BGL significantly increased the abundance of the zonula occludens-1-(ZO-1) in the jejunal epithelium upon ETEC challenge (p < 0.05). BGL supplementation also increased the number of S-phase cells and the number of sIgA-positive cells, but significantly decreased the number of total apoptotic cells in the jejunal epithelium upon ETEC challenge (p < 0.05). Moreover, BGL significantly increased the duodenal catalase (CAT) activity and the ileal total superoxide dismutase (T-SOD) activity in the ETEC-challenged pigs (p < 0.05). Importantly, BGL significantly decreased the expression levels of critical inflammation related proteins such as the tumor necrosis factor-α (TNF-α), interlukin-6 (IL-6), myeloid differentiation factor 88 (MyD88), and nuclear factor-κB (NF-κB) in the jejunal and ileal mucosa upon ETEC challenge (p < 0.05). BGL also elevated the propanoic acid content and the abundance of Lactobacillus and Bacillus in the colon upon ETEC challenge (p < 0.05). These results suggested BGL could alleviate the ETEC-induced intestinal epithelium injury, which may be associated with suppressed inflammation and improved intestinal immunity and antioxidant capacity, as well as the improved intestinal macrobiotic.

Manno-oligosaccharide attenuates inflammation and intestinal epithelium injury in weaned pigs upon enterotoxigenic Escherichia coli K88 challenge.

To explore the effect of manno-oligosaccharide (MOS) on intestinal health in weaned pigs upon enterotoxigenic Escherichia coli K88 (ETEC) challenge, thirty-two male weaned pigs were randomly assigned into four groups. Pigs fed with a basal diet or basal diet containing MOS (0·6 g/kg) were orally infused with ETEC or culture medium. Results showed that MOS significantly elevated the digestibility of crude protein and gross energy in both ETEC-challenged and non-challenged pigs (P < 0·05). MOS also elevated serum concentrations of IgA and IgM (P < 0·05), but decreased serum concentrations of TNF-α, IL-1ß and IL-6 (P < 0·05) in ETEC-challenged pigs. Interestingly, MOS increased villus height and the ratio of villus height:crypt depth in duodenum and ileum (P < 0·05). MOS also increased duodenal sucrase and ileal lactase activity in ETEC-challenged pigs (P < 0·05). MOS decreased the abundance of E. coli, but increased the abundance of Lactobacillus, Bifidobacterium and Bacillus in caecum (P < 0·05). Importantly, MOS not only elevated the expression levels of zonula occludens-1 (ZO-1), claudin-1 and GLUT-2 in duodenum (P < 0·05) but also elevated the expression levels of ZO-1, GLUT-2 and L-type amino acid transporter-1 in ileum (P < 0·05) upon ETEC challenge. These results suggested that MOS can alleviate inflammation and intestinal injury in weaned pigs upon ETEC challenge, which was associated with suppressed secretion of inflammatory cytokines and elevated serum Ig, as well as improved intestinal epithelium functions and microbiota.

Dietary copper/zinc-loaded montmorillonite improved growth performance and intestinal barrier and changed gut microbiota in weaned piglets.

The effects of copper/zinc-loaded montmorillonite (Cu/Zn-Mt) on growth performance, intestinal barrier and gut microbiota of weaned pigs were investigated in the present study. A total of 108 piglets (Duroc × Landrace × Yorkshire; 6.36 kg; weaned at 21 ± 1 d age) were used in this experiment. The pigs were randomly assigned to three treatments with six replicates, six pigs in each replicate. The three treatments were as follows: (1) control group: basal diet; (2) Cu/Zn-Mt group: basal diet supplemented with 39 mg/kg Cu and 75 mg/kg Zn as Cu/Zn-Mt; and (3) Cu +Zn +Mt group: basal diet supplemented with the mixture of copper sulphate, zinc sulphate and montmorillonite (equivalent to the copper and zinc in the Cu/Zn-Mt treatment). The results indicated that, compared with the pigs from control group, average daily gain and gain: feed ratio were increased and the faecal score on days 7 and 14 after weaning was decreased by supplementation of Cu/Zn-Mt; intestinal transepithelial electrical resistance (TER) and expressions of tight junction protein claudin-1 and zonula occludens-1 were increased, and intestinal permeability of fluorescein isothiocyanate-dextran 4 kDa was decreased by supplementation with Cu/Zn-Mt. According to the Illumina-based sequencing results, Cu/Zn-Mt supplementation increased the relative abundance of core bacteria (Lactococcus, Bacillus) at genus level and decreased the potentially pathogenic bacteria (Streptococcus and Pseudomonas) in colon of weaned piglets. However, the piglets fed with the mixture of copper sulphate, zinc sulphate and montmorillonite showed no effects in above parameters in comparison with the pigs from control group. In conclusion, dietary Cu/Zn-Mt could improve growth performance, decrease the diarrhoea and improve intestinal barrier and bacterial communities of weaned pigs. The results indicated that 'loading' of montmorillonite with Zn and Cu changed not only its chemical but also its nutritional properties.

Effects of dietary supplementation of compound enzymes on performance, nutrient digestibility, serum antioxidant status, immunoglobulins, intestinal morphology and microbiota community in weaned pigs.

The objective of this study was to evaluate the effects of compound enzymes (CE) (containing per g 375 U amylase, 2500 U protease, 4000 U xylanase and 150 U ß-glucanase) on performance, nutrient digestibility, serum antioxidant status, immunoglobulins, intestinal morphology, volatile fatty acids contents and microbiota community in weaned pigs. Seventy-two pigs (Duroc × Landrace × Yorkshire, weaned at d 28) with an average body weight of 8.49 ± 0.87 kg were allotted into two treatments with six replicate pens per treatment (three barrows and three gilts per pen) according to sex and body weight in a randomised complete block design. The treatments contained a corn-soybean meal-barley basal diet (CON) or a basal diet supplemented with 1000 mg CE/kg (CE). The study was divided into phase 1 (d 1 to 14) and 2 (d 15 to 35). The average daily gain was increased (p < 0.05) in pigs fed CE in phase 2 and overall (d 1 to 35) compared with CON. These pigs had greater (p ≤ 0.05) serum IgA, IgG, superoxide dismutase and catalase contents, as well as tended to increase serum IgM content and apparent total tract digestibility (ATTD) of organic matter in phase 1 compared with CON. In phase 2, pigs supplemented with CE showed greater (p < 0.01) ATTD of dry matter, organic matter, crude protein and gross energy compared with CON. These pigs also had increased (p < 0.05) IgA, IgG, IgM, superoxide dismutase contents, and decreased (p < 0.05) malondialdehyde content in serum compared with CON. Moreover, pigs fed CE had higher (p < 0.05) villus height and villus height to crypt depth ratio in ileum, and tended to increased acetic acid content in colon compared with CON. Furthermore, pigs fed CE had increased (p < 0.05) relative abundance of Firmicutes at phylum level, Lactobacillales at order level, Lactobacillaceae at family level, Bacilli at class level, Lactobacillus at genus level in caecum and colon, as well as lower (p < 0.05) relative abundance of Bacteroidetes at phylum level, Bacteroidales at the order level, Bacteroidia at class level, Clostridium_sensu_stricto_6 at genus level in colon compared with CON. In conclusion, dietary inclusion of compound enzymes could effectively improve nutrient digestibility, serum antioxidant status, immunoglobulin, gut morphology, microbiota community, and therefore improve performance in weaned pigs.

Effects of tea tree oil supplementation on growth performance, antioxidant capacity, immune status and microbial community in weaned pigs.

The objective of this study was to determine whether dietary tea tree oil (TTO) supplementation could effectively replace the antibiotics through modulating the antioxidant capacity and intestinal microbiota profile, and then decreasing the diarrhoea incidence and improving the growth performance of weaned pigs. A total of 216 weaned pigs with initial body weights (BW) of 9.19 ± 1.86 kg were randomly allocated to three dietary treatments in a completely randomised design. The dietary treatments included a corn-soybean meal basal diet (CON) without any antibiotics, and two experimental diets formulated by adding 75 mg/kg aureomycin (AGP) or 100 mg/kg TTO into the basal diet, respectively. Pigs fed the TTO diet showed greater gain to feed ratio (p < 0.05) than those fed CON and AGP diets during d 0-14 and d 14-28. Both dietary TTO and AGP supplementation tended to increase the average daily gain of weaned pigs during d 14-28 (p = 0.06) and the overall 28-d period (p = 0.07), and significantly reduced (p < 0.05) the diarrhoea incidence during d 0-14 compared with the CON treatment. In addition, dietary TTO supplementation improved the apparent total tract digestibility of dry matter and ether extract (p < 0.05), and increased (p < 0.05) the propionate and butyrate concentrations in faecal samples of weaned pigs. Moreover, pigs fed the TTO diet showed greater total antioxidant capacity, greater superoxide dismutase and interleukin-10 concentrations, and lower malondialdehyde concentration in serum than those fed the CON diet (p < 0.05). Furthermore, pigs fed the TTO diet demonstrated greater relative abundance of Clostridiaceae_1, while those fed the AGP diet exhibited greater relative abundance of Lactobacillaceae at family level. In conclusion, dietary TTO supplementation could improve growth performance in weaned pigs, which could be mainly attributed to the benefits on nutrient digestibility, antioxidative capacity and microbial community profile.

Dietary supplementation of plant essential oil improves growth performance, intestinal morphology and health in weaned pigs.

The present study was conducted to explore the effect of plant essential oil (PEO) on growth performance, intestinal morphology and health in weaned pigs. Twenty-four weaned pigs were assigned into four groups fed with a basal diet (CON) or basal diet containing PEO at a concentration of 50 (PEO50), 100 (PEO100) or 200 ppm (PEO200). After 21 days, pigs were slaughtered and blood and tissue samples were collected. Result showed that PEO200 group significantly increased the average daily gain (ADG) compared with CON group (p < .05). Moreover, PEO supplementation significantly improved the digestibility of DM (p < .05). However, it significantly decreased the serum triglyceride and cholesterol concentration (p < .05). Interestingly, PEO supplementation significantly increased the activity of sucrase in the duodenal and the activity of lactase in the jejunal mucosa (p < .05). Moreover, PEO supplementation improved the growth of intestinal mucosa. As compared to the CON group, the jejunum and ileum villus height were significantly elevated in the PEO200 group (p < .05). Importantly, the expression levels of critical genes associated with nutrient transportation (i.e., GLUT2 and SGLT1) and barrier function (occludin) were significantly elevated in the PEO200 group (p < .05). Moreover, the PEO100 and PEO200 group had higher propionic acid concentration and higher total bacterial gene copies in colon digesta than the CON group (p < .05) respectively. These results not only suggest that PEO has a positive role in the regulation of growth and intestinal health in weaned pigs, but also offer a potential candidate substituting the conventionally used antibiotics in the livestock industry.

Effect of fibre sources on performance, serum parameters, intestinal morphology, digestive enzyme activities and microbiota in weaned pigs.

This study was conducted to evaluate the effects of wheat bran (WB) as insoluble fibre source, and sugar beet pulp (SBP) as soluble fibre source, on performance, serum parameters and intestinal health in weaned pigs. A total of 90 weaned pigs (BW: 7.33 ± 1.24 kg) were randomly assigned to three dietary treatments: (1) a control diet (CON) based on corn and soybean meal; (2) CON + 6% WB; (3) CON + 6% SBP. Each treatment had five replicate pens with six pigs per pen. The experimental period was divided into two phases (d 0 to 14 and d 14 to 28). Pigs in group WB tended to have greater avarage daily gain than those in group SBP. Compared with CON, SBP reduced (p < 0.05) the apparent total tract digestibility (ATTD) of dry matter, organic matter (OM), gross energy and neutral detergent fibre on d 14 and 28, while WB decreased (p < 0.05) the ATTD of OM on d 28. Pigs in group SBP had higher (p < 0.05) glucagon-like peptide 1 concentration than the other groups on d 14 and 28. The villus height to crypt depth (V:C) ratio of duodenum and jejunum in pigs fed diet WB were greater (p < 0.05) than in group SBP. The WB increased (p < 0.05) the V:C ratio of ileum compared with CON or SBP. Compared with SBP, WB increased (p < 0.05) the sucrase activity in the duodenum. Moreover, pigs in WB had higher (p < 0.05) activities of maltase and sucrase in the jejunum compared with CON or SBP. The abundances of Ruminococcaceae and Prevotellaceae were increased (p < 0.05) in WB, while the Lachnospiraceae abundance was increased (p < 0.05) in SBP. WB increased (p < 0.05) concentrations of acetate, butyrate and total short-chain fatty acids (SCFA), while SBP increased (p < 0.05) concentrations of acetate and total SCFA when compared with CON. In conclusion, WB was beneficial to performance in weaned pigs by improving morphology, enzyme activities and microbiota when compared with SBP, highlighting that effects of fibre depends on the fibre sources.

Moderate tetrabasic zinc chloride supplementation improves growth performance and reduces diarrhea incidence in weaned pigs.

OBJECTIVE: Two experiments were conducted to evaluate tetrabasic zinc chloride (TBZC) on the health of weaned pigs, and to determine the optimal supplemental concentrations and whether dietary TBZC could replace the pharmacological concentrations of dietary zinc oxide (ZnO) to improve growth performance and decrease Zn excretion in weaned pigs. METHODS: In Exp. 1, 180 weaned pigs (8.92 ± 1.05 kg BW) were randomly assigned to 1 of 5 treatments, including the basal diet containing 125 mg/kg zinc sulfate (ZnSO4), and the basal diet with 1,200, 1,800, 2,400, or 3,000 mg/kg TBZC supplementation. In Exp. 2, 240 weaned pigs (7.66 ± 1.09 kg BW) were randomly assigned to 1 of 5 treatments, including a negative control diet without Zn supplementation (NC), a positive control diet (2,250 mg/kg ZnO), and 3 experimental diets with different concentrations of TBZC supplementation (1,000, 1,250 and 1,500 mg/kg). RESULTS: In Exp. 1, the average daily gain (ADG), feed efficiency (G:F) and diarrhea incidence responded quadratically (p&lt;0.01) as the TBZC supplemental concentrations increased, and pigs fed 1,200 and 1,800 mg/kg TBZC showed the best growth performance. Moreover, 1,800 mg/kg TBZC supplementation showed the greatest (p&lt;0.01) total antioxidant capacity (T-AOC) and glutathione peroxidase (GSH-Px) activities in liver of pigs. Histopathological examination revealed lesions in heart, liver, lung and kidney, and mild or severe histological lesions mainly occurred with the supplementation of 2,400 and 3,000 mg/kg TBZC. In Exp. 2, 1,000 and 1,250 mg/kg TBZC supplementation in diets significantly (p&lt;0.01) increased ADG and G:F of weaned pigs, reduced Zn excretion in feces, and had no effect on diarrhea-reducing compared to 2,250 mg/kg ZnO supplementation. CONCLUSION: TBZC is a potential alternative to ZnO. The recommended concentration of TBZC in weaned pig diets is 1,000 to 1,250 mg/kg.

Hot melt extruded-based nano zinc as an alternative to the pharmacological dose of ZnO in weanling piglets.

OBJECTIVE: This study was conducted to investigate the effects of hot-melt extruded ZnO nano-particles (HME-ZnO) as an alternative for P-ZnO on growth performance, nutrient digestibility, Zn bioavailability, intestinal microbiota, and intestinal morphology of weanling pigs. METHODS: A total of 450 piglets (Landrace×Yorkshire×Duroc) were randomly allotted to five treatments based on initial body weight and sex. The experimental diets were fed in a meal form as phase 1 from d 0 to 14 and phase 2 from d 15 to 28. Treatments were the control diet without ZnO supplementation, the diet containing 2,500 ppm Zn as ZnO, and three diets containing 500, 1,000, or 2,500 ppm Zn as HME-ZnO. RESULTS: The overall result showed a higher (p<0.01) average daily gain in weanling pigs fed ZnO-supplemented diets in comparison to the control diet. There was a decrease (p<0.01) in fecal score in the ZnO-supplemented diets. Dietary supplementation of ZnO improved (p<0.05) crude protein digestibility. The weanling pigs fed the P-ZnO diet had a lower (p<0.01) Zn digestibility in the feces than HME-ZnO supplemented treatments. Weanling pigs fed diets supplemented with ZnO had greater (p<0.05) Lactobacillus spp. populations and lower Clostridium spp. (p<0.05) and Coliforms (p<0.01) populations in the ileum. Weanling pigs fed diets supplemented with increasing concentrations of HME-ZnO linearly decreased Clostridium spp. (p<0.05) and Coliforms (p<0.01) in the ileum. Lower (p<0.05) Clostridium spp. and Coliforms counts in the colon were observed in pigs fed with ZnO-supplemented diets. Weanling pigs fed diets supplemented with ZnO showed a greater (p<0.01) villus height in the duodenum. CONCLUSION: Dietary supplementation of HME-ZnO and P-ZnO showed a potential to improve the digestibility of protein, intestinal Coliform and Clostridium, villus height in duodenum and ileum. Moreover, HME-ZnO showed a higher Zn digestibility compared with P-ZnO.

Changes of porcine gut microbiota in response to dietary chlorogenic acid supplementation.

Chlorogenic acids (CGA), the most abundant natural polyphenol present in human diet and plants, have attracted considerable research interest because of their broad bioactivities including the antimicrobial activity. However, little is known about their influences on intestinal bacterial communities. Here, we described a response in intestinal microbiome to CGA using a porcine model. Twenty-four weaned pigs were allotted to two groups and fed with a basal diet or a basal diet containing 1000 mg/kg CGA. Results showed that CGA significantly increased the length of the small intestine (P < 0.05) and enhanced the activity of diamine oxidase (DAO) and the concentration of MHC-II in the jejunal and ileal mucosa (P < 0.05). Moreover, the acetate concentration in ileum and cecum digesta, and the propionate and butyrate concentrations in the cecum digesta, were significantly elevated by CGA (P < 0.05). Interestingly, CGA significantly increased the total 16S rRNA gene copies and bacterial alpha diversity in the cecum (P < 0.05). The relative abundance of bacteria from phyla Firmicutes and Bacteroidetes was increased in the cecum digesta (P < 0.05), whereas the abundance of bacteria from phylum Protebacteria was decreased by CGA (P < 0.05). Importantly, pigs on CGA-containing diet had higher abundance of Lactobacillus spp., Prevotella spp., Anaerovibrio spp., and Alloprevotella spp. in the cecum (P < 0.05). Not only did our study suggest a synergic response of intestinal barrier function and microbiota to the CGA, but the result will also contribute to understanding of the mechanisms behind the CGA-modulated gut health.

Combined supplementation of Lactobacillus fermentum and Pediococcus acidilactici promoted growth performance, alleviated inflammation, and modulated intestinal microbiota in weaned pigs.

BACKGROUND: Probiotics are important for pigs to enhance health and intestinal development, which are potential alternative to antibiotics. Many studies have reported the functions of single bacterial strain as probiotic on the animals. In this study, we evaluated effects of combined probiotics on growth performance, inflammation and intestinal microbiota in weaned pigs. One hundred and eight pigs, weaned at 28 day old (7.12 ± 0.08 kg), were randomly divided into the 3 dietary treatments with 6 pens and 6 pigs per pen (half male and half female). The experimental period lasted for 28 days and treatments were as follows: i. CONTROL: basal diet; ii. Antibiotic: the basal diet plus 75 mg· kg- 1 chlortetracycline; and iii. Probiotics: basal diet plus 4% compound probiotics. RESULTS: Supplementation probiotics improved average daily gain over the entire 28 days (P < 0.01) and feed efficiency in the last 14 days (P < 0.05) compared with the other two groups. Both probiotics and antibiotic supplementation decreased concentrations of serum pro-inflammatory cytokines interleukin-6 (P < 0.05) and interferon-γ (P < 0.01). Probiotics group had greater abundance of Lactobacillus in the caecal digesta and Firmicutes in the colonic digesta, while both probiotics and antibiotic supplementation inhibited Treponema_2 and Anaerovibrio in the caecal digesta. Caecal acetic and propionic acid (P < 0.05) of probiotics group were higher than the other two groups, whereas concentrations of colonic lactic acid and propionic acid (P < 0.05) of antibiotic group were lower than control and probiotics groups. CONCLUSIONS: These findings suggest that combined supplementation of Lactobacillus fermentum and Pediococcus acidilactici regulate the gut health and improve the host ADG and F/G by decreasing serum pro-inflammatory factors (IL-6, IFN-γ), promoting beneficial bacteria (Lactobacillus in the caecal digesta and Firmicutes in the colonic digesta), enhancing production of short chain fatty acids, and inhibiting pathogens (Treponema_2, Anaerovibrio in the caecal digesta).

Pathogenicity and immunogenicity of attenuated porcine epidemic diarrhea virus PC22A strain in conventional weaned pigs.

BACKGROUND: Porcine epidemic diarrhea virus (PEDV) causes diarrhea in all ages of pigs with 50-100% mortality rates in neonatal piglets. In the United States, inactivated and subunit PEDV vaccines for pregnant sows are available, but fail to induce sufficient protection in neonatal piglets farrowed from PEDV naïve sows. A safe and efficacious live attenuated vaccine that can prime mucosal immune responses is urgently needed. In this study, we evaluated the safety and efficacy of two attenuated PEDV vaccine candidates, the emerging non-S INDEL PEDV strain PC22A at the 100th cell culture passage level - Clone no. 4 (P100C4) and at the 120th passage level (P120), in weaned pigs. RESULTS: Four groups of 40-day-old weaned pigs were inoculated orally with PEDV PC22A-P3 (virulent), -P100C4, -P120, and mock, respectively, and challenged with the P3 virus at 24 days post-inoculation (dpi). After inoculation, P3 caused diarrhea in all pigs with a high level of fecal viral RNA shedding. P100C4 and P120 did not cause diarrhea in pigs, although viral RNA was detected in feces of all pigs, except for one P100C4-inoculated pig. Compared with the P120 group, P3- and P100C4-inoculated pigs had higher serum PEDV-specific IgG and viral neutralizing (VN) antibody (Ab) titers at 14 dpi. After the challenge, no pigs in the P3 group but all pigs in the P100C4, P120, and mock groups had diarrhea. Compared with the P120 group, pigs in the P100C4 group had a more rapid decline in fecal PEDV RNA shedding titers, higher titers of serum PEDV-specific IgG, IgA, and VN Abs, and higher numbers of intestinal IgA Ab-secreting cells. CONCLUSIONS: PEDV PC22A P100C4 and P120 were fully attenuated in weaned pigs but failed to elicit protection against virulent P3 challenge. P100C4 induced higher PEDV-specific antibody responses than P120 post inoculation resulting in a greater anamnestic response post challenge. Therefore, P100C4 potentially could be tested as a priming vaccine or be further modified using reverse genetics. It also can be administered in multiple doses or be combined with inactivated or subunit vaccines and adjuvants as a PEDV vaccination regimen, whose efficacy can be tested in the future.

Amelioration of Enterotoxigenic Escherichia coli-Induced Intestinal Barrier Disruption by Low-Molecular-Weight Chitosan in Weaned Pigs is Related to Suppressed Intestinal Inflammation and Apoptosis.

Enterotoxigenic Escherichia coli (ETEC) infection destroys the intestinal barrier integrity, in turn, disrupting intestinal homoeostasis. Low-molecular-weight chitosan (LMWC) is a water-soluble chitosan derivative with versatile biological properties. Herein, we examined whether LMWC could relieve ETEC-induced intestinal barrier damage in weaned pigs. Twenty-four weaned pigs were allotted to three treatments: (1) non-infected control; (2) ETEC-infected control; and (3) ETEC infection + LMWC supplementation (100 mg/kg). On day 12, pigs in the infected groups were administered 100 mL of ETEC at 2.6 × 109 colony-forming units/mL to induce intestinal barrier injury. Three days later, serum samples were obtained from all pigs, which were then slaughtered to collect intestinal samples. We evidenced that LMWC not only increased (P < 0.05) the occludin protein abundance but also decreased (P < 0.05) the interleukin-6, tumour necrosis factor-α and mast cell tryptase contents, and the apoptotic epithelial cell percentages, in the small intestine of ETEC-infected pigs. Furthermore, LMWC down-regulated (P < 0.05) the small intestinal expression levels of critical inflammatory- and apoptotic-related genes, such as Toll-like receptor 4 (TLR4) and tumour necrosis factor receptor 1 (TNFR1), as well as the intra-nuclear nuclear factor-κB (NF-κB) p65 protein abundance, in the ETEC-infected pigs. Our study indicated a protective effect of LMWC on ETEC-triggered intestinal barrier disruption in weaned pigs, which involves the repression of intestinal inflammatory responses via blocking the TLR4/NF-κB signalling pathway and the depression of epithelial cell death via TNFR1-dependent apoptosis.

Effects of low-protein diet on the intestinal morphology, digestive enzyme activity, blood urea nitrogen, and gut microbiota and metabolites in weaned pigs.

This study investigated the effects of low-protein diet supplemented with Lysine (Lys), Methionine (Met), Threonine (Thr), and Tryptophan (Trp) on small intestine morphology, enzyme activity, blood urea nitrogen, and gut microbiota and metabolites in weaned piglets. Eighteen weaned pigs weighing an average of 9.57 kg received one of three treatments: a normal protein diet with 20% crude protein (CP, diet [NP]), a moderately reduced protein diet with 17% CP (MP), or a low-protein diet with 14% CP (LP). All three diets were supplemented with Lys, Met, Thr and Trp to meet essential amino acid requirements for post-weaned piglets according to the NRC (2012). Following a 45 d study period, piglets on the LP and MP diets demonstrated atrophic small intestinal morphology, with decreased villus heights and lower ratios of villus height to crypt depth (p < 0.05); pepsin activity in the stomach was also reduced in these two groups (p < 0.05). Increased plasma cholesterol and decreased blood urea nitrogen presented in the MP and LP groups compared with the NP group (p < 0.05). Overall, gastrointestinal hormones were not affected by dietary protein levels with the exception of reduced somatostatin levels in the MP and LP groups. Jejunum and colon microbiota were not affected at either the phyla or genera level in any of the diets. Colonic ammonia nitrogen concentration was reduced in MP and LP groups. Dietary protein level had no effect on short chain fatty acids or biogenic amines. Our data suggest that reducing dietary protein levels by 3% (MP) or 6% (LP) in weaned pigs has the potential to decrease nitrogen emissions and impaired digestive capacity. Therefore, dietary protein level cannot be reduced by more than 3% in consideration of maladaptive changes to small intestinal morphology and pepsin activity in weaned piglets.

Frequency of diarrhoea-associated viruses in swine of various ages in Hungary.

Enteric viral diseases of swine are one of the most frequent disorders causing huge economic losses in pork production. After the reappearance of an emerging enteropathogen, porcine epidemic diarrhoea virus (PEDV) in Hungary in 2016, an extensive survey was initiated in an attempt to identify diarrhoea-related porcine viruses, including adeno-, astro-, boca-, calici-, circo-, corona-, kobu-, rota- and Torque teno viruses. A total of 384 faecal samples collected during a twoyear period from diarrhoeic and asymptomatic pigs of various ages in 17 farms were screened by conventional and real-time PCR methods. Half of the samples contained at least one examined virus with the dominance of kobuvirus (55.1%) followed by bocaviruses (33.2%) and rotavirus groups A and C together (20.9%), while coronaviruses including PEDV were not found in this set of samples. Statistical analysis showed a highly significant difference (P < 0.0001) in the frequency of single infections compared to mixed ones with the exception of weaned pigs, in which group additionally most viruses were detected. The results of this study suggest that the complexity of this disease may vary with age, which makes the prevention of diarrhoea a challenge, especially in weaned pigs.