Influence of high- and low-fermentable dietary fibres in sows' diet on the colostrum potential against Clostridioides difficile toxin-induced effects in IPEC-J2 cells

Sow colostrum has been reported to protect the IPEC-J2 cells and piglet colon tissues from detrimental effect of Clostridioides difficile toxins. Since dietary fibre can influence the colostrum composition in sows, we hypothesised that it can also differentially affect the colostrum potential against C. difficile toxin-induced effects in IPEC-J2. IPEC-J2 were incubated with colostrum from sows fed either high-fermentable sugar beet pulp (SBP) or low-fermentable lignocellulose (LNC) fibres and in combination with the toxins and analysed by trans-epithelial electrical resistance (TEER) and cell viability using propidium iodide in flow cytometry. Toxins drastically decreased the integrity of IPEC-J2. Colostrum from the sows fed either SBP or LNC exerted protective effect against toxins on IPEC-J2 integrity and this effect was numerically superior in the SBP group. Differences in the percentages of TEER between different treatments were noted after 2 h (p = 0.043), 3 h (p = 0.017) and 4 h (p = 0.017) of incubation and a tendency for differences was noted after 5 h of incubation (p = 0.071). Colostrum from either SBP- or LNC-fed sows did not protect the IPEC-J2 from toxin-induced death. Colostrum of the sows fed either high-fermentable or low-fermentable fibres has a potential to protect IPEC-J2 from the loss of integrity, which may be important in protection from C. difficile-infection development in neonatal piglets

Developing Gut Microbiota Exerts Colonisation Resistance to Clostridium (syn. Clostridioides) difficile in Piglets

Clostridium (syn. Clostridioides) difficile is considered a pioneer colonizer and may cause gut infection in neonatal piglets. The aim of this study was to explore the microbiota-C. difficile associations in pigs. We used the DNA from the faeces of four sows collected during the periparturient period and from two to three of their piglets (collected weekly until nine weeks of age) for the determination of bacterial community composition (sequencing) and C. difficile concentration (qPCR). Furthermore, C. difficile-negative faeces were enriched in a growth medium, followed by qPCR to verify the presence of this bacterium. Clostridium-sensu-stricto-1 and Lactobacillus spp. predominated the gut microbiota of the sows and their offspring. C. difficile was detected at least once in the faeces of all sows during the entire sampling period, albeit at low concentrations. Suckling piglets harboured C. difficile in high concentrations (up to log10 9.29 copy number/g faeces), which gradually decreased as the piglets aged. Enrichment revealed the presence of C. difficile in previously C. difficile-negative sow and offspring faeces. In suckling piglets, the C. difficile level was negatively correlated with carbohydrate-fermenting bacteria, and it was positively associated with potential pathogens. Shannon and richness diversity indices were negatively associated with the C. difficile counts in suckling piglets. This study showed that gut microbiota seems to set conditions for colonisation resistance against C. difficile in the offspring. However, this conclusion requires further research to include host-specific factors

Porcine Colostrum Protects the IPEC-J2 Cells and Piglet Colon Epithelium against Clostridioides (syn. Clostridium) difficile Toxin-Induced Effects

Clostridioides difficile toxins are one of the main causative agents for the clinical symptoms observed during C. difficile infection in piglets. Porcine milk has been shown to strengthen the epithelial barrier function in the piglet’s intestine and may have the potential to neutralise clostridial toxins. We hypothesised that porcine colostrum exerts protective effects against those toxins in the IPEC-J2 cells and in the colon epithelium of healthy piglets. The IPEC-J2 cells were treated with either the toxins or porcine colostrum or their combination. Analyses included measurement of trans-epithelial electrical resistance (TEER), cell viability using propidium iodide by flow cytometry, gene expression of tight junction (TJ) proteins and immune markers, immunofluorescence (IF) histology of the cytoskeleton and a TJ protein assessment. Colon tissue explants from one- and two-week-old suckling piglets and from five-week-old weaned piglets were treated with C. difficile toxins in Ussing chamber assays to assess the permeability to macromolecules (FITC-dextran, HRP), followed by analysis of gene expression of TJ proteins and immune markers. Toxins decreased viability and integrity of IPEC-J2 cells in a time-dependent manner. Porcine colostrum exerted a protective effect against toxins as indicated by TEER and IF in IPEC-J2 cells. Toxins tended to increase paracellular permeability to macromolecules in colon tissues of two-week-old piglets and downregulated gene expression of occludin in colon tissues of five-week-old piglets (p = 0.05). Porcine milk including colostrum, besides other maternal factors, may be one of the important determinants of early immune programming towards protection from C. difficile infections in the offspring

Porcine and Chicken Intestinal Epithelial Cell Models for Screening Phytogenic Feed Additives—Chances and Limitations in Use as Alternatives to Feeding Trials

Numerous bioactive plant additives have shown various positive effects in pigs and chickens. The demand for feed additives of natural origin has increased rapidly in recent years to support the health of farm animals and thus minimize the need for antibiotics and other drugs. Although only in vivo experiments can fully represent their effect on the organism, the establishment of reliable in vitro methods is becoming increasingly important in the goal of reducing the use of animals in experiments. The use of cell models requires strict control of the experimental conditions so that reliability and reproducibility can be achieved. In particular, the intestinal porcine epithelial cell line IPEC-J2 represents a promising model for the development of new additives. It offers the possibility to investigate antioxidative, antimicrobial, anti- or pro-proliferative and antiviral effects. However, the use of IPEC-J2 is limited due to its purely epithelial origin and some differences in its morphology and functionality compared to the in vivo situation. With regard to chickens, the development of a reliable intestinal epithelial cell model has attracted the attention of researchers in recent years. Although a promising model was presented lately, further studies are needed to enable the standardized use of a chicken cell line for testing phytogenic feed additives. Finally, co-cultivation of the currently available cell lines with other cell lines and the development of organoids will open up further application possibilities. Special emphasis was given to the IPEC-J2 cell model. Therefore, all publications that investigated plant derived compounds in this cell line were considered. The section on chicken cell lines is based on publications describing the development of chicken intestinal epithelial cell models

Fiber Composition in Sows’ Diets Modifies Clostridioides difficile Colonization in Their Offspring

Dietary fiber has a potential to modulate the gut microbiota in sows. We hypothesized that a maternal diet rich in either high- or low-fermentable fiber during gestation and lactation influences Clostridioides difficile gut colonization in suckling piglets. Twenty sows were fed gestation and lactation diets enriched with either high-fermentable sugar beet pulp (SBP) or low-fermentable lignocellulose (LNC) fibers. C. difficile, toxin B (TcdB), fecal score, microbial abundance (16S-rDNA sequencing) and metabolites were measured in the feces from the sows and their piglets. C. difficile concentration was higher in piglets from the sows fed LNC than SBP along the study (P ≤ 0.05). Higher prevalence of C. difficile was noted in three-week-old piglets from sows fed LNC vs. SBP (45% vs. 0%, P = 0.001). TcdB prevalence was higher in six-day-old piglets from the sows fed LNC vs. SBP (60% vs. 17%, P = 0.009). In sows, fecal microbial metabolites were higher in SBP than LNC, while C. difficile concentration showed no difference. Higher microbial diversity Shannon index was noted in sows from SBP vs. LNC one week before parturition and at the parturition (P ≤ 0.05). Piglets from SBP vs. LNC tended to have higher microbial diversity Shannon index at two and three weeks of age. Diets enriched with high-fermentable fiber compared to low-fermentable fiber in sows reduced C. difficile colonization in their piglets. Susceptibility to colonization by C. difficile in neonatal piglets can be modulated by the sows’ diet, supporting the hypothesis of the early microbial programming in the offspring and the importance of the sow-piglet couple

Clostridioides difficile-mesocolonic oedema in neonatal suckling piglets develops regardless of the fibre composition in sow’s diets

Maternal dietary factors have been reported to influence Clostridioides difficile colonisation in the offspring. Twenty suckling piglets from sows fed diets supplemented with high-fermentable sugar beet pulp (SBP) or low-fermentable lignocellulose (LNC) fibres during gestation and lactation were dissected in the first week after birth. Postmortem analysis included clinical mesocolon and faecal scoring, concentration of C. difficile and respective toxins in colon digesta and faeces, immunoglobulins in serum and inflammatory markers in serum and colon tissues. Sow colostrum was assessed for nutrients, immunoglobulins and biogenic amines. Toxin-neutralising IgG antibodies were measured in colostrum and serum of the sows, and in colon digesta and serum of the piglets. Mesocolonic oedema of different severity was present in most of the piglets from both sows’ feeding groups. Concentrations of C. difficile, toxins and calprotectin in colon digesta and faecal contents did not differ between the study piglets. Calprotectin correlated positively with mesocolon score (rho = 413, P = 0.07). Piglets from sows fed LNC vs SBP tended to have higher IgA (P = 0.089), IgG (P = 0.053), total Ig (P = 0.053), albumin (P = 0.075) and total protein content (P = 0.007) in serum. Colon tissues of piglets from the SFB vs LNC had upregulated expression of ZO-1 (P = 0.021), PCNA (P = 0.015) and TGF-β (P = 0.014). Titers of anti-toxin-IgG-antibodies in serum and colostrum and in piglet colon digesta and serum did not differ between sows from both dietary groups, but they all showed strong positive correlations. In conclusion, dietary sugar beet pulp or lignocellulose fed to sows did not influence the concentrations of C. difficile and toxins titers in colon digesta and faeces of neonatal piglets

Dietary fiber and its role in performance, welfare, and health of pigs

Dietary fiber (DF) is receiving increasing attention, and its importance in pig nutrition is now acknowledged. Although DF for pigs was frowned upon for a long time because of reductions in energy intake and digestibility of other nutrients, it has become clear that feeding DF to pigs can affect their well-being and health. This review aims to summarize the state of knowledge of studies on DF in pigs, with an emphasis on the underlying mode of action, by considering research using DF in sows as well as suckling and weaned piglets, and fattening pigs. These studies indicate that DF can benefit the digestive tracts and the health of pigs, if certain conditions or restrictions are considered, such as concentration in the feed and fermentability. Besides the chemical composition and the impact on energy and nutrient digestibility, it is also necessary to evaluate the possible physical and physiologic effects on intestinal function and intestinal microbiota, to better understand the relation of DF to animal health and welfare. Future research should be designed to provide a better mechanistic understanding of the physiologic effects of DF in pigs

Biodiversity of soil bacteria exposed to sub-lethal concentrations of phosphonium-based ionic liquids: Effects of toxicity and biodegradation

Little is known about the effect of ionic liquids (ILs) on the structure of soil microbial communities and resulting biodiversity. Therefore, we studied the influence of six trihexyl(tetradecyl)phosphonium ILs (with either bromide or various organic anions) at sublethal concentrations on the structure of microbial community present in an urban park soil in 100-day microcosm experiments. The biodiversity decreased in all samples (Shannon's index decreased from 1.75 down to 0.74 and OTU's number decreased from 1399 down to 965) with the largest decrease observed in the microcosms spiked with ILs where biodegradation extent was higher than 80%. (i.e. [P66614][Br] and [P66614][2,4,4]). Despite this general decrease in biodiversity, which can be explained by ecotoxic effect of the ILs, the microbial community in the microcosms was enriched with Gram-negative hydrocarbon-degrading genera e.g. Sphingomonas. It is hypothesized that, in addition to toxicity, the observed decrease in biodiversity and change in the microbial community structure may be explained by the primary biodegradation of the ILs or their metabolites by the mentioned genera, which outcompeted other microorganisms unable to degrade ILs or their metabolites. Thus, the introduction of phosphonium-based ILs into soils at sub-lethal concentrations may result not only in a decrease in biodiversity due to toxic effects, but also in enrichment with ILs-degrading bacteria