Effects of variety and storage duration on the nutrient digestibility and the digestible and metabolisable energy content of maize fed to growing pigs.

The objective of this research was to determine the effects of variety and storage duration on the nutrient digestibility and the digestible (DE) and metabolisable (ME) energy content in maize when fed to growing pigs. Four maize varieties (LS1, LS2, LS3 and LS4) were hand-harvested from the same growing area in China in early October of 2012. The samples were sun dried to about 14% moisture content and then stored in the warehouse of the Fengning Pig Experiment Base at China Agricultural University for 0, 3 or 10 months. Twenty-four barrows of about 33 kg body weight were used and allotted to a completely randomised block design with four diets and six replicate pigs per diet. Pigs were individually housed in metabolic crates. The four experimental diets were formulated by mixing 96.8% of each variety of maize with 3.2% vitamins and minerals. A 5-day collection period followed a 7-day diet acclimation period. The results indicated that the DE and ME contents of maize and the apparent total tract digestibility (ATTD) of organic matter (OM), dry matter, gross energy (GE), neutral detergent fibre, acid detergent fibre (ADF), crude protein (CP) and ether extract (EE) were significantly (p < 0.05) influenced by maize variety and storage duration. With an extension of storage duration from 0 to 10 months, the DE and ME of maize and the ATTD of OM, GE, ADF, CP and EE changed in a quadratic manner (p < 0.05), and 3 months of storage exceeded 0 months of storage by 1.84%, 1.43%, 0.31%, 0.32%, 15.37%, 2.11% and 5.02%, respectively. The DE, ME of maize and the ATTD of OM, GE, ADF, CP and EE decreased by 3.67%, 6.00%, 0.97%, 1.40%, 30.54%, 3.92% and 20.93%, respectively, at 10 months of storage compared to 3 months of storage. No interaction was observed between maize variety and storage duration in DE and ME contents in maize. In conclusion, under the conditions of this study, most of the nutrient digestibility and the DE and ME contents of maize increased from 0 to 3 months and decreased from 3 to 10 months.

Protection of mice against enterotoxigenic E. coli by immunization with a polyvalent enterotoxin comprising a combination of LTB, STa, and STb.

Currently available enterotoxigenic Escherichia coli (ETEC) vaccines are based on colonization factors and/or the heat-labile enterotoxin B subunit (LTB). However, the induction of antitoxic responses against heat-stable enterotoxin a (STa) and b (STb) has merit as these two poorly immunogenic toxins are frequently associated with ETEC strains. In this study, we genetically constructed a trivalent enterotoxin fusion protein (STa-LTB-STb, abbreviated to SLS) in an effort to develop a single toxoid containing these three enterotoxins for vaccination against ETEC. Mutagenesis at one disulfide-bridge-forming cysteine in STa led to a dramatic reduction in the STa toxicity of SLS; however, the fusion peptide retained the STb-associated toxicity. Immunization of mice with SLS protein elicited significant antibody responses to LTB, STa, and STb. Significantly, the mice antisera were able to neutralize the biological activity of both STa and STb. In the experiment to assess the protective effect of SLS immunization, the mortality of mice receiving SLS was significantly lower than their control cohorts (P < 0.01) after intraperitoneal challenge with ETEC. These results show that the trivalent fusion enterotoxin SLS has the potential to serve as a useful toxin-based vaccine against ETEC-induced diarrheal disease via a single immunogen.

Lactobacillus reuteri I5007 Modulates Intestinal Host Defense Peptide Expression in the Model of IPEC-J2 Cells and Neonatal Piglets.

Modulation of the synthesis of endogenous host defense peptides (HDPs) by probiotics represents a novel antimicrobial approach for disease control and prevention, particularly against antibiotic-resistant infections in human and animals. However, the extent of HDP modulation by probiotics is species dependent and strain specific. In the present study, The porcine small intestinal epithelial cell line (IPEC-J2) cells and neonatal piglets were used as in-vitro and in-vivo models to test whether Lactobacillus reuteri I5007 could modulate intestinal HDP expression. Gene expressions of HDPs, toll-like receptors, and fatty acid receptors were determined, as well as colonic short chain fatty acid concentrations and microbiota. Exposure to 108 colony forming units (CFU)/mL of L. reuteri I5007 for 6 h significantly increased the expression of porcine ß-Defensin2 (PBD2), pBD3, pBD114, pBD129, and protegrins (PG) 1-5 in IPEC-J2 cells. Similarly, L. reuteri I5007 administration significantly increased the expression of jejunal pBD2 as well as colonic pBD2, pBD3, pBD114, and pBD129 in neonatal piglets (p < 0.05). This was probably associated with the increase in colonic butyric acid concentration and up-regulating expression of Peroxisome Proliferator-Activated Receptor-γ (PPAR-γ) and G Protein-Coupled Receptor 41 (GPR41) (p < 0.05), but not with stimulation of Pattern-Recognition Receptors. Additionally, supplementation with L. reuteri I5007 in the piglets did not affect the colonic microbiota structure. Our findings suggested that L. reuteri I5007 could modulate intestinal HDP expression and improve the gut health of neonatal piglets, probably through the increase in colonic butyric acid concentration and the up-regulation of the downstream molecules of butyric acid, PPAR-γ and GPR41, but not through modifying gut microbiota structure.

Chicken egg yolk antibodies (IgY) modulate the intestinal mucosal immune response in a mouse model of Salmonella typhimurium infection.

This study determined the effects of chicken egg yolk antibodies (IgY) on immune responses in the intestinal mucosal of mice infected with Salmonella typhimurium. Sixty, 28-day-old mice were divided into 4 groups and treated with streptomycin or sterile water for 2days followed by 1day without treatment. The control group was unchallenged whereas the mice in the other three groups were treated twice with 10(9)CFUmL(-1)S. typhimurium. For the next 3days, control mice continued to receive no treatment whereas the mice in the remaining three groups were orally administered with 20mgmL(-1) of specific IgY, 20mgmL(-1) of nonspecific IgY or PBS. S. typhimurium activated gut-associated lymphoid tissue, increasing the release of IFN-γ and TNF-α in the mucosa and increased the number of activated T-lymphocytes and cytotoxic T-γδ. Specific IgY attenuated the increase in IFN-γ and TNF-α and the decrease in IL-10. S. typhimurium induced mobilization of CD8(+) and CD8(+) TCRγδ T cells in the epithelium and CD4(+) and CD8(+) T cells in the lamina propria reflecting an inflammatory process that was attenuated by IgY. These results suggest that specific IgY modulates intestinal mucosal immune responses during a S. typhimurium infection.

Functions of Antimicrobial Peptides in Gut Homeostasis.

Antimicrobial peptides (AMPs), produced by several species including bacteria, insects, amphibians and mammals as well as by chemical synthesis and genetically engineered microorganisms, are of great importance in maintaining normal gut homeostasis. AMPs exhibit a broad spectrum of antimicrobial activity and inhibit microbial cells by interaction with their membranes or by other mechanisms, such as inhibition of cell-wall synthesis or suppression of nucleic acid or protein synthesis. In addition to their direct antimicrobial functions, they have multiple roles in the stabilization of epithelial barrier integrity and function as potent immune regulators. The fate of AMPs in vivo is poorly understood, prompting the need for studying AMPs pharmacokinetics. This review summarizes the current knowledge about the basic biology of AMPs and discusses the features of AMPs in gut homeostasis and their relative mechanisms of action.

Intestinal microbiota succession and immunomodulatory consequences after introduction of Lactobacillus reuteri I5007 in neonatal piglets.

Seventy-two, suckling piglets, obtained from 9 litters standardized to 8 piglets, were assigned to 1 of 3 treatments (n = 24) to compare short-term, early administration with intermittent, longer-term administration of Lactobacillus reuteri I5007. The treatments were a control (given a placebo of 0.1% peptone water from day 1 to 5) or treatments in which 1.7 × 1010 CFU L. reuteri was administrated either daily for 4 days starting on day 1 or every 4th day from day 1 to 17. Five piglets per treatment were killed at 3 time points (day 7, 14 and 21). Denaturing Gradient Electrophoresis of ileal digesta revealed an increase in the presence of L. reuteri I5007 and Clostridium lentocellum (on day 14 and 21) in the every 4th-day treatment and Actinobacillus porcinus (on day 7 and 14) in both L. reuteri treatments, while reducing the abundance of E. coli on day 21 in the every 4th-day treatment. Real-time qPCR of ileal digesta showed an increase in Bifidobacterium spp. on day 14 for both L. reuteri I5007 treatments. An increase in the concentration of lactic acid and a lower pH was observed in the first 4-day treatment on day 7 and the every 4th day treatment on day 14. The relative abundance of mRNA for TGF-ß was increased while that for IFN-γ was decreased in the mesenteric lymph nodes of piglets treated with L. reuteri every 4th day. In conclusion, early intervention with L. reuteri increases the presence of beneficial bacteria and decreases the presence of undesirable microbes in the lower gastrointestinal tract. The changes appear to be mediated by altering the intestinal pH through lactic acid production resulting in favorable bacterial species colonization. A prolonged duration of treatment (i.e. every 4th day) would appear to be superior to treatment only during the first 4 days.

Alternatives to antibiotics as growth promoters for use in swine production: a review.

In the past two decades, an intensive amount of research has been focused on the development of alternatives to antibiotics to maintain swine health and performance. The most widely researched alternatives include probiotics, prebiotics, acidifiers, plant extracts and neutraceuticals such as copper and zinc. Since these additives have been more than adequately covered in previous reviews, the focus of this review will be on less traditional alternatives. The potential of antimicrobial peptides, clay minerals, egg yolk antibodies, essential oils, eucalyptus oil-medium chain fatty acids, rare earth elements and recombinant enzymes are discussed. Based on a thorough review of the literature, it is evident that a long and growing list of compounds exist which have been tested for their ability to replace antibiotics as feed additives in diets fed to swine. Unfortunately, the vast majority of these compounds produce inconsistent results and rarely equal antibiotics in their effectiveness. Therefore, it would appear that research is still needed in this area and that the perfect alternative to antibiotics does not yet exist.

Effect of wheat distillers' grains with solubles and a feed flavour on performance and carcass traits of growing-finishing pigs fed wheat and canola meal based diets.

Forty-eight crossbred pigs were assigned to one of six dietary treatments in a 6 x 2 (treatment x sex) factorial arrangement. Diets were based on wheat and canola meal and were formulated to contain 0%, 4.9%, 9.7%, 14.6% or 19.4% wheat distillers' dried grains with solubles (DDGS) during the growing period and 0%, 4.0%, 8.1%, 12.1% and 16.1% wheat DDGS during the finishing period. The addition of wheat DDGS was made at the expense of both wheat and canola meal. A feed flavour was added to the diet in which wheat DDGS supplied 100% of the supplementary protein. Over the entire experimental period (21.5-112.2 kg), increasing the level of wheat DDGS resulted in a linear decrease in weight gain and feed conversion ratio. Feed intake was linearly reduced by inclusion of wheat DDGS during the growing period (21.5-57.4 kg) but not the finishing period (57.4-112.2 kg). Increasing the level of wheat DDGS in the diet resulted in a linear decline in carcass value index and lean yield while loin fat linearly increased. The addition of a flavour to the diet in which DDGS supplied 100% of the supplementary protein had no effect on performance or carcass traits.

Effects of the antimicrobial peptide cecropin AD on performance and intestinal health in weaned piglets challenged with Escherichia coli.

This study was conducted to determine the effects of the antimicrobial peptide cecropin on performance and intestinal health in piglets. Newly weaned barrows were randomly assigned to one of three treatments (n=8), including a corn-soybean basal diet or similar diets supplemented with antibiotics (100 mg/kg kitasamycin plus 800 mg/kg colistin sulfate) or 400 mg/kg cecropin AD. On day 13, all piglets were orally challenged with 10(9)CFU/mL of Escherichia coli K88. On day 19, all piglets were euthanized and sampled. Before challenge, piglets fed antibiotics had greater weight gain, feed efficiency, nitrogen and energy retention than the control (P<0.05). E. coli challenge decreased weight gain, feed intake and feed efficiency for the control piglets (P<0.05) but not for the antibiotic or cecropin AD treated piglets. The incidence of diarrhea post-challenge in the antibiotic and cecropin AD treatments decreased compared with the control piglets. The total viable counts of cecal E. coli were lower while the Lactobacilli counts were higher in the antibiotic and cecropin AD treatments compared with the control (P<0.05). Cecropin AD treatment decreased total aerobes while increasing total anaerobes in the ileum (P<0.05). A higher villus height to crypt depth ratio in the jejunum and ileum as well as a deeper crypt depth in the jejunum and higher villus height in the ileum were observed in piglets fed antibiotics or cecropin AD compared with control piglets (P<0.05). Piglets fed the control diet had lower levels of secretory IgA in their jejunum and lower serum IgA, IgG, interleukin-1ß and interleukin-6 compared with the other treatments (P<0.05). Overall, these data suggest that cecropin AD enhances pig performance through increasing immune status and nitrogen and energy retention as well as reducing intestinal pathogens in weaned piglets.