Acinetobacter in fresh faecal samples is related to sperm motility: A case report in Piétrain boars housed on different deep-litter bedding materials.

The swine gastrointestinal tract contains a great variety of microbes, forming a complex and dynamic ecosystem. Various internal and external factors (e.g. age, breed and diet) may influence its composition. This study aimed to investigate the gut microbial diversity of German Piétrain boars housed on different deep-litter bedding materials (regional wood shavings, linen, hemp, spelt husks, and wood shavings) via 16S-rDNA sequencing. Additionally, short-chain fatty acids were analysed using gas chromatography. Fresh faecal samples (n = 80) from 40 Piétrain boars were collected twice during the trial. Although it can be assumed that boars ingest bedding orally, no differences in the microbiome composition could be found. The main phyla were Firmicutes and Bacteroides. Acinetobacter was identified as a biomarker for sperm quality differences (total sperm motility) in breeding boars.

Inclusion of a Bacillus-based probiotic in non-starch polysaccharides-rich broiler diets.

This study examined the effects of a 3-strain Bacillus-based probiotic (BP; Bacillus amyloliquefaciens and two Bacillus subtilis) in broiler diets with different rye levels on performance, mucus, viscosity, and nutrient digestibility. We distributed 720 one-d-old female broilers into 72 pens and designed nine diets using a 3 × 3 factorial approach, varying BP levels (0, 1.2 × 106, and 1.2 × 107 CFU/g) and rye concentrations (0, 200, 400 g/kg). On d 35, diets with 200 or 400 g/kg rye reduced broiler weight gain (BWG). Diets with 400 g/kg rye had the highest FCR, while rye-free diets had the lowest (p ≤0.05). Adding BP increased feed intake and BWG in weeks two and three (p ≤0.05). It should be noted that the overall performance fell below the goals of the breed. Including rye in diets reduced the coefficient of apparent ileal digestibility (CAID) for protein, ether extract (EE), calcium, phosphorus, and all amino acids (p ≤0.05). Rye-free diets exhibited the highest CAID for all nutrients, except for methionine, EE, and calcium, while diets with 400 g/kg of rye demonstrated the lowest CAID (p ≤0.05). BP in diets decreased phosphorus CAID (p ≤0.05). Diets containing 1.2 × 107 CFU/g (10X) of BP exhibited higher CAID of methionine than the other two diets (p ≤0.05). Diets containing 10X of BP showed higher CAID of cysteine than diets with no BP (p ≤0.05). Ileal viscosity increased as the inclusion level of rye in the diets increased (p ≤0.05). The ileal concentration of glucosamine in chickens fed diets with 400 g/kg of rye was higher than in those fed diets with no rye (p ≤0.05). Furthermore, ileal galactosamine concentrations were elevated in diets with 200 and 400 g/kg of rye when compared to rye-free diets (p ≤0.05). However, BP in diets had no impact on ileal viscosity, galactosamine, or glucosamine (p > 0.05). In conclusion, the applied Bacillus strains appeared to have a limited capacity to produce arabinoxylan-degrading enzymes and were only partially effective in mitigating the negative impacts of rye arabinoxylans on broilers.

Performance and nutrient digestibility of growing pigs fed highly or low fermentable coarse or finely ground fibre-rich feedstuffs.

Dietary fibre is mainly classified according to its chemical characteristics but structure and particle size of fibre-rich feedstuff can also be decisive for digestion and performance. So far, only few studies investigated this in pigs. This experiment aimed to compare coarse and finely ground dried hemp plants and apple pomace regarding performance and ileal and total tract nutrient digestibility of growing pigs. Coarse or finely ground apple pomace or dried hemp plants were added to the diet of 56 nine weeks old growing pigs (DanBred x Duroc), housed in flat decks with each 2 animals. The growing pigs received the experimental diets for three weeks while performance was recorded. Eight pigs per group were sacrificed and digesta and organ tissue sampled. The stomach health was evaluated by visually scoring of the mucosa integrity. Apparent ileal (AID) and total tract digestibility (ATTD) were calculated using titanium dioxide as marker. Statistical analyses were performed using two-way ANOVA (p < 0.05). The highest feed intake (fibre particle size, p = 0.018) and bodyweight gain (fibre particle size, p = 0.018; fibre source x particle size interaction, p = 0.040), was observed in animals fed finely ground apple pomace, while the feed conversion ratio was 8-12% lower in pigs fed finely ground fibre sources (p = 0.012). No differences in stomach mucosa integrity were detected between the groups. The relative pancreas (p = 0.045), stomach (p < 0.001), and jejunum (p = 0.010) weights were higher in animals fed diets containing apple pomace. In contrast, the relative liver, caecum and colon weights were not affected by fibre source or particle size. The AID of protein and amino acids was not affected, while ATTD was increased by fibre source (hemp vs. apple pomace) reducing faecal nitrogen excretion. The AID of calcium was increased when diets contained apple pomace (p < 0.001), while zinc AID and ATTD were enhanced when diets contained dried hemp (p = 0.016; p = 0.016, respectively). Our results suggest that the structure as well as the chemical characteristics should be considered in a future fibre evaluation system in pigs.

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.

Effects of dietary zinc chloride hydroxide and zinc methionine on the immune system and blood profile of healthy adult horses and ponies.

The effects of dietary zinc on the immune function of equines have not been evaluated in detail so far. In the present study, eight healthy adult ponies and two healthy adult horses were fed a diet supplemented with either zinc chloride hydroxide or zinc methionine in six feeding periods of four weeks each (according to maintenance zinc requirement, 120 mg zinc/kg dry matter, and 240 mg zinc/kg dry matter, for both dietary zinc supplements, respectively). All animals received the six diets, with increasing amounts of zinc chloride hydroxide in the feeding periods 1-3, and with increasing amounts of zinc methionine in the feeding periods 4-6. At the end of each feeding period, blood samples were collected for a blood profile and the measurement of selected immune variables. Increasing dietary zinc chloride hydroxide doses increased the glutathione concentrations in the erythrocyte concentrate and the glutathione peroxidase activity in the erythrocyte lysate, decreased the numbers of total leukocytes and granulocytes in the blood, and also decreased the interleukin-2 concentrations in the plasma of the animals. The dietary supplementation of increasing doses of zinc methionine enhanced the mitogen-stimulated proliferative activity of peripheral blood mononuclear cells, and decreased the glutathione concentrations in the erythrocyte concentrate and the glutathione peroxidase activity in the plasma of the animals. The percentage of blood monocytes with oxidative burst after in vitro stimulation with E. coli decreased with increasing dietary zinc concentrations, independently of the zinc compound used. The blood profile demonstrated effects of the zinc supplements on the red blood cells and the bilirubin metabolism of the horses and ponies, which require further investigation. Overall, high doses of dietary zinc modulate the equine immune system, for the most part also depending on the zinc compound used.

Glutamine supplementation moderately affects growth, plasma metabolite and free amino acid patterns in neonatal low birth weight piglets.

Low birth weight (LBW) neonates show impaired growth compared with normal birth weight (NBW) neonates. Glutamine (Gln) supplementation benefits growth of weaning piglets, while the effect on neonates is not sufficiently clear. We examined the effect of neonatal Gln supplementation on piglet growth, milk intake and metabolic parameters. Sow-reared pairs of newborn LBW (0·8-1·2 kg) and NBW (1·4-1·8 kg) male piglets received Gln (1 g/kg body mass (BM)/d; Gln-LBW, Gln-NBW; n 24/group) or isonitrogenous alanine (1·22 g/kg BM/d; Ala-LBW; Ala-NBW; n 24/group) supplementation at 1-5 or 1-12 d of age (daily in three equal portions at 07:00, 12:00 and 17:00 by syringe feeding). We measured piglet BM, milk intake (1, 11-12 d), plasma metabolite, insulin, amino acid (AA) and liver TAG concentrations (5, 12 d). The Gln-LBW group had higher BM (+7·5%, 10 d, P = 0·066; 11-12 d, P < 0·05) and milk intake (+14·7%, P = 0·015) than Ala-LBW. At 5 d, Ala-LBW group had higher plasma TAG (+34·7%, P < 0·1) and lower carnosine (-22·5%, P < 0·05) than Ala-NBW and Gln-LBW, and higher liver TAG (+66·9%, P = 0·029) than Ala-NBW. At 12 d, plasma urea was higher (+37·5%, P < 0·05) with Gln than Ala supplementation. Several proteinogenic AA in plasma were lower (P < 0·05) in Ala-NBW v. Gln-NBW. Plasma arginine was higher (P < 0·05) in Gln-NBW v Ala-NBW piglets (5, 12 d). Supplemental Gln moderately improved growth and milk intake and affected lipid metabolism in LBW piglets and AA metabolism in NBW piglets, suggesting effects on intestinal and liver function.

Gut Microbiota of Pigs Metabolizes Extracts of Filipendula ulmaria and Orthosiphon aristatus-Herbal Remedies Used in Urinary Tract Disorders.

Urinary tract infections influence the mortality rate in pigs and are linked to extensive antibiotic usage in the farm industry. Filipendula ulmaria (L.) Maxim. and Orthosiphon aristatus (Blume) Miq. are widespread medicinal plants traditionally used to treat urinary tract disorders. As their preparations are orally administered, the metabolism of their constituents by gut microbiota before absorption should be considered. Until now, no experiments had been performed to describe the biotransformation of tthose plants' extracts by animal gut microbiota. The study evaluates the influence of pig intestinal microbiota on the structure of active compounds in flowers of F. ulmaria and leaves of O. aristatus. The incubations of the extracts with piglet gut microbiota were performed in anaerobic conditions, and the samples of the batch culture were collected for 24 h. In F. ulmaria, the main metabolites were quercetin and kaempferol, which were products of the deglycosylation of flavonoids. After 24 h incubation of O. aristatus extract with the piglet gut microbiota, 2 main metabolites were observed. One, tentatively identified as 3-(3-dihydroxyphenyl)propionic acid, is likely the primary metabolite of the most abundant depsides and phenolic acids. The results confirm the formation of the compounds with anti-inflammatory and diuretic activity in the microbiota cultures, which might suggest F. ulmaria and O. aristatus for treating urinary tract disorders in piglets. Based on the similarities of human and pig gut microbiota, the pig model can help estimate the metabolic pathways of natural products in humans.

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.

Performance, health, bacterial metabolites and intestinal histomorphology in does and growing rabbits fed diets with increasing lignocellulose-to-cellulose proportions.

Adequate levels of dietary fibre can reduce the risk of intestinal health disorders in rabbits after weaning. Therefore, it was the aim of the study to investigate the impact of different lignocellulose and cellulose addition to rabbits' diets on performance and intestinal traits in lactating does and weaned rabbits. A total of 60 rabbit does (4.41 ± 0.45 kg body weight) were fed isonitrogenous diets with lignocellulose and cellulose inclusion rates [%] of 0/8, 2/6, 4/4, 6/2 and 8/0, resulting in dietary lignin-to-cellulose ratios of 0.26, 0.30, 0.32, 0.34 and 0.40, respectively. The diets were fed from 4 d ante partum (a.p.) until day 28 post partum (p.p. weaning). After weaning, 40 young rabbits from each feeding group were fed the same diets for 12 d, when 10 rabbits per group were sacrificed. Body weight gain (BWG), feed intake and feed conversion ratio were obtained. Gastrointestinal content and gut tissue were obtained from the growing rabbits and pH, short-chain fatty acids (SCFA) concentration and gut morphology were investigated. Digestive disorders were scored, and the sanitary risk index was calculated. Does fed the highest lignocellulose addition increased their feed intake [g · kg BW-1 · d-1] in the first two weeks p.p. (p < 0.001) and increased BWG (p < 0.001) but without impact on their offsprings' performance. Weaned rabbits fed the highest lignocellulose inclusion showed a reduced feed intake and increased sanitary risk index. Additionally, a higher number of rabbits showed intestinal disorders for a short time (1-2 d, p < 0.001). In the colon content, a higher pH and a lower dry matter content were observed in this group. Caecal contents of 40-d old rabbits showed lower total SCFA, acetate and propionate and higher i-valerate and L-lactate concentrations after feeding diets with high lignocellulose contents. The diet did not influence the acetate-to-propionate ratio. No histomorphological changes were observed in the jejunum, while crypt depth and width were reduced in the caecum of weaned rabbits fed diets with lignocellulose addition. It can be concluded that different proportions of lignocellulose to cellulose addition to rabbits' diets have an impact on health and performance with differences between does and weaned rabbits. While a high lignocellulose inclusion proved beneficial for does, for weaned rabbits, a lower addition should be preferred.

Comparative studies of urolithins and their phase II metabolites on macrophage and neutrophil functions.

PURPOSE: Ellagitannins are high molecular weight polyphenols present in high quantities in various food products. They are metabolized by human and animal gut microbiota to postbiotic metabolites-urolithins, bioavailable molecules of a low molecular weight. Following absorption in the gut, urolithins rapidly undergo phase II metabolism. Thus, to fully evaluate the mechanisms of their biological activity, the in vitro studies should be conducted for their phase II conjugates, mainly glucuronides. The aim of the study was to comparatively determine the influence of urolithin A, iso-urolithin A, and urolithin B together with their respective glucuronides on processes associated with the inflammatory response. METHODS: The urolithins obtained by chemical synthesis or isolation from microbiota cultures were tested with their respective glucuronides isolated from human urine towards modulation of inflammatory response in THP-1-derived macrophages, RAW 264.7 macrophages, PBMCs-derived macrophages, and primary neutrophils. RESULTS: Urolithin A was confirmed to be the most active metabolite in terms of LPS-induced inflammatory response inhibition (TNF-α attenuation, IL-10 induction). The observed strong induction of ERK1/2 phosphorylation has been postulated as the mechanism of its action. None of the tested glucuronide conjugates was active in terms of pro-inflammatory TNF-α inhibition and anti-inflammatory IL-10 and TGF-ß1 induction. CONCLUSION: Comparative studies of the most abundant urolithins and their phase II conjugates conducted on human and murine immune cells unambiguously confirmed urolithin A to be the most active metabolite in terms of inhibition of the inflammatory response. Phase II metabolism was shown to result in the loss of urolithins' pharmacological properties.

A Preliminary Survey of the Distribution of Segmented Filamentous Bacteria in the Porcine Gastrointestinal Tract.

Segmented filamentous bacteria (SFB) are present in various animal species including pigs. The aim of this work was to analyze the occurrence of SFB in different parts of the gastrointestinal tract of piglets of different ages. A total of 377 DNA extracts from stomach, jejunum, ileum, cecum and colon digesta, and from feces collected on different time points, originating from 155 animals, were screened by qPCR method with primers specific for the SFB. SFB sequences were detected in 74 of 377 samples (19.6%) from 155 animals in total. SFB were most abundant in ileum (50.0%), cecum (45.0%), and colon (37.0%), followed by feces (14.6%). SFB prevalence in sows was 12.9% (13/101) and 75.9% (41/54) in individual piglets. Of the 41 SFB-positive piglets, only two samples were from pre-weaning animals, while the rest of samples originated from post-weaning piglets. SFB sequences are abundant in post-weaning piglets, but not in suckling or adult animals. They are most abundant in the ileum and cecum of pigs. Further studies are warranted to reveal the role of SFB in pigs.

Effects of dietary cellobiose on the intestinal microbiota and excretion of nitrogen metabolites in healthy adult dogs.

In order to evaluate the potential prebiotic effects of cellobiose, 10 healthy adult research beagle dogs received a complete diet containing 0, 0.5 and 1 g cellobiose/kg bodyweight (BW)/day. At the end of each feeding period, faeces, urine and blood of the dogs were collected. The results demonstrated a significant increase of faecal lactate concentrations, indicating a bacterial fermentation of cellobiose in the canine intestine. Along with this, a dose-dependent linear increase of the relative abundance of Lactobacillaceae in the faeces of the dogs was observed (p = 0.014). In addition, a dose-dependent increase (p < 0.05) of Alloprevotella, Bacteroides and Prevotella, and a linear decrease for unidentified Lachnospiraceae (p = 0.011) was observed when cellobiose was added to the diet, although the relative abundance of these genera was low (<1%) among all groups. The faecal pH was not affected by dietary cellobiose. Cellobiose seemed to modulate the excretion of nitrogen metabolites, as lower concentrations of phenol (p = 0.034) and 4-ethylphenol (p = 0.002) in the plasma of the dogs were measured during the supplementation periods. Urinary phenols and indoles, however, were not affected by the dietary supplementation of cellobiose. In conclusion, cellobiose seems to be fermented by the intestinal microbiota of dogs. Although no effect on the faecal pH was detected, the observed increase of microbial lactate production might lower the pH in the large intestine and consecutively modulate the intestinal absorption of nitrogen metabolites. Also, the observed changes of some bacterial genera might have been mediated by increased intestinal lactate concentrations or a higher relative abundance of lactobacilli. Whether these results could be considered as a prebiotic effect and used as a dietetic strategy in diseased animals to improve gut function or hepatic and renal nitrogen metabolism should be evaluated in future studies.

Effect of essential oils or saponins alone or in combination on productive performance, intestinal morphology and digestive enzymes' activity of broiler chickens.

This study was carried out to evaluate the effect of essential oils or saponins alone or in combination on productive performance, intestinal morphology, and digestive enzymes' activity in broilers. Four hundred one-day-old male broiler chickens were randomly assigned to four treatments, each with 5 replicates. The birds were fed experimental diets contained either no phytogenic feed additives (control) or were supplemented with 25.0 mg/kg of an essential oil blend from star anise, rosemary, thyme and oregano (essential oils); 46.0 mg/kg of a Quillaja saponin blend (saponins); or a combination of both phytogenic preparations (essential oils plus saponins). The experiment lasted for 42 days. Body weight, feed intake and weight gain were recorded weekly, and the feed conversion ratio was calculated, throughout the experiment. Intestinal morphology and digestive enzymes' activity were determined at the end of the experiment. It was found that the body weight was increased by all dietary supplements at the end of the experiment. There was no change in performance parameters among the treatments during the starter period. At the grower period and the overall experiment, the weight gain of birds was higher in all supplemented groups compared to the control, but the feed conversion was better in the essential oils with saponins treatment. Supplementation of essential oils or saponins alone or in combination increased the villus height and the villus/crypt ratio of the intestine. There was an increase in trypsin, chymotrypsin and elastase activities in the essential oils plus saponins group only. The obtained results indicate that the phytogenic feed additives, especially essential oils with saponins, can be considered as new promising agents in promoting the growth performance of broilers. The effect could be related to its improving impact in intestinal development and increasing the protease enzymes' activity, resulting in increased protein digestibility and absorption of dietary nutrients.

Effects of cereal and protein source on performance, apparent ileal protein digestibility and intestinal characteristics in weaner piglets.

Rye and rapeseed meal (RSM) are alternatives to wheat and soybean meal (SBM) in pig nutrition. The inclusion of rye and RSM instead of wheat and SBM may increase dietary fibre content in the feed, which may affect digestive physiology and intestinal function. However, research on the combined feeding of rye and RSM in piglets is still scarce. The aim of this study was to investigate the effects of rye and RSM containing feed on growth performance, protein and amino acid digestibility, physical characteristics of the digesta and the intestinal morphology in weaner piglets. A total of 88 piglets aged 28 d were randomly allotted to four different feeding groups at weaning: wheat/SBM, wheat/RSM, rye/SBM, rye/RSM. Cereals were included at 48%, SBM at 25% and RSM at 30%. Body weight gain and feed intake were recorded weekly. After 33 d, piglets were euthanised for assessment of digesta and intestinal morphology. Rye did not affect performance, whilst RSM resulted in reduced weight gain (p = 0.024) and feed intake (p = 0.037). Apparent ileal digestibility of crude protein and amino acids was decreased by rye and RSM (p < 0.05). Rye caused an acidification (p = 0.045) and increased viscosity (p = 0.013) of small intestinal digesta, a lower dry matter content of jejunal and colonic digesta (p < 0.05) and an increased relative weight of the colon tissue (p < 0.001). In conclusion, performance was not reduced when rye was used as an alternative to wheat in diets for weaner piglets. The inclusion of RSM instead of SBM to piglets' diet lowered animal performance under our conditions. Further investigations are warranted to characterise the quality of rapeseed meal and the optimum inclusion level in the diet for young pigs.

Impact of the dietary inclusion of dried food residues on the apparent nutrient digestibility and the intestinal microbiota of dogs.

The use of food residues for animal nutrition might imply ecological and economic advantages; however, their effects as a potential ingredient have not yet been evaluated in dogs. In the present study, four diets with 0, 5, 10 and 15% dried food residues (DFR), derived from hotel catering, were fed to 10 healthy adult dogs. At the end of each three-week feeding period, faeces and blood were collected. The apparent nutrient digestibility was calculated by the dietary inclusion of titanium dioxide as an inert marker. The results demonstrated that the apparent crude protein digestibility and ether extract digestibility decreased with increasing amounts of DFR in the diets (p < 0.05). In addition, an increase of the faecal concentrations of acetic acid, propionic acid, n-butyric acid and total short-chain fatty acids (SCFA) was observed (p < 0.05). Faecal ammonium and lactate concentrations, as well as plasma phenol and indole concentrations, were not linearly affected by the dietary inclusion of DFR. The relative abundance of Fusobacteria in the faeces of the dogs decreased, and the relative abundance of Actinobacteria and Bacteroidetes increased with increasing amounts of DFR in the diets (p < 0.05). In conclusion, the DFR seemed to be intensively fermented by the intestinal microbiota of the dogs, as indicated by the increased faecal SCFA concentrations and the shifts in the composition of the faecal microbiota. Dietary inclusion levels of up to 5% can be recommended based on our results, as the observed lower apparent crude protein and ether extract digestibility might limit the use of food residues for dogs at higher amounts.

In vitro conjugation kinetics of AmpC, broad spectrum and extended-spectrum beta-lactamase-producing Escherichia coli donors and various Enterobacteriaceae recipients.

BACKGROUND: Extended spectrum beta-lactamase (ESBL)-producing enterobacteria pose a major hazard to public health. Due to the possibility of genetic transfer, ESBL genes might spread to pathogenic enterobacterial strains. Thus, information on possible genetic transfer between enterobacteria is of high interest. It was therefore the aim of this in vitro study to screen the capacity of a wide range of Enterobacteriaceae for differences in conjugation at different time points with five ESBL-producing Escherichia coli strains. RESULTS: Conjugation frequencies for five potential E. coli donor strains producing the enzymes CTX-M-1, CTX-M-15, SHV-12, TEM-1, TEM-52 and CMY-2, and six potential recipient strains commonly detected in the gastrointestinal tract of poultry (E. coli, Serratia marcescens subsp. marcescens, Enterobacter cloacae, Salmonella (S.) enterica serovar Typhimurium and Proteus mirabilis) were obtained. Different combinations of donor and recipient strains were co-incubated for between 0 and 22 h and spread on selective agar. Conjugation frequencies were calculated as transconjugants per donor. Some donor and recipient strain combinations did not perform plasmid transfer within 22 h. Hence, the recipient Proteus mirabilis did not accept plasmids from any of the given donors and the E. coli ESBL10716 donor was unable to transfer its plasmid to any recipient. Enterobacter cloacae only accepted the plasmids from the donors E. coli ESBL10708 and E. coli ESBL10716 while E. coli ESBL10708 did not transfer its plasmid to Serratia marcescens subsp. marcescens. E. coli IMT11716 on the other hand did not perform conjugation with the donor E. coli ESBL10689. The remaining mating pairs differed in conjugation frequency, ranging from 10- 5 to 10- 9 transconjugants/donor. The earliest conjugation events were detected after 4 h. However, some mating pairs turned positive only after 22 h of coincubation. CONCLUSION: A suitable mating pair for future in vivo studies to combat transfer of antibiotic resistance to pathogenic bacteria in broiler chicken was determined. The results of this study also suggest that the kinetic of conjugation differs between mating pairs and is independent of species origin. This should be considered when performing conjugation experiments.

Dietary Supplementation of dl-Methionine Potently Induces Sodium-Dependent l-Methionine Absorption in Porcine Jejunum Ex Vivo.

BACKGROUND: Methionine is an essential amino acid (AA) with many fundamental roles. Humans often supplement l-Met, whereas dl-Met and dl-2-hydroxy-4-(methylthio)butanoic acid (dl-HMTBA) are more frequently used to supplement livestock. OBJECTIVES: The study aimed to investigate whether dietary Met source alters the absorptive capacity for Met isomers in the small intestine of piglets. METHODS: A total of 27 male 10-wk-old piglets in 3 feeding groups received a diet supplemented with 0.21% dl-Met, 0.21% l-Met, or 0.31% dl-HMTBA to meet the Met + cystine requirement. After ≥10 d, absorptive fluxes of d-Met or l-Met were measured at a physiological concentration of 50 µM and a high concentration of 5 mM in duodenum, middle jejunum, and ileum ex vivo. Data were compared by 2-factor ANOVA. RESULTS: Across diets, fluxes of both Met isomers at both tested concentrations increased from duodenum to ileum by a factor of ∼2-5.5 (P < 0.05). Pigs supplemented with dl-Met had greater (P < 0.085) absorptive fluxes at 50 µM l-Met (0.50, 2.07, and 3.86 nmol · cm-2 · h-1) and d-Met (0.62, 1.41, and 1.19 nmol · cm-2 · h-1) than did pigs supplemented with dl-HMTBA (l-Met: 0.28, 0.76, and 1.08 nmol · cm-2 · h-1; d-Met: 0.34, 0.58, and 0.64 nmol · cm-2 · h-1) in duodenum, jejunum, and ileum, respectively. Only in jejunum of dl-Met-fed pigs, fluxes at 50 µM l-Met were reduced by the omission of luminal Na+ (from 3.27 to 0.86 nmol · cm-2 · h-1; P < 0.05) and by a cocktail of 22 luminal AAs (to 1.05 nmol · cm-2 · h-1; P < 0.05). CONCLUSIONS: Dietary supplementation of dl-Met increases the efficiency of l-Met and d-Met absorption at physiologically relevant luminal Met concentrations along the small intestine of pigs, including a very prominent induction of an Na+-dependent transport system with preference for l-Met in the mid-jejunum. Dietary supplementation with dl-Met could be a promising tool to improve the absorption of Met and other AAs.

Distinct patterns of microbial metabolic fingerprints in sows and their offspring: a pilot study.

Microbial metabolism and growth in the intestinal tract depend on the composition of substrates present in the digesta and their ability to be metabolised by the microorganisms. The aim of this pilot study was to characterise potential hindgut microbial activity during perinatal period in sows and their offspring. Rectal samples from three sows (1-3 weeks before and after birth) and three of their piglets (1-5 weeks after birth), were subjected to assays using BIOLOG GEN III microplates to produce metabolic fingerprints for each animal. The number of metabolised substrates of the sow hindgut microbiota was stable during the pregnancy and lactation periods, as assessed by the richness index. In piglets, the richness was stable during the suckling period and at beginning of weaning, however, it decreased when the piglets were 5 weeks old (P ≤ 0.05). Analysis of associations between the sows and the piglets and the microbial metabolic potential showed that microbial metabolism was strongly associated with the catabolism of carbohydrates especially in sows. Only 5-week-old weaned piglets clustered together with the sows regarding the microbial catabolism of substrates, but not suckling piglets. The association analyses clustered all the piglets in two groups distinctive for litter. The analysis of metabolic fingerprints via microbial growth with different substrates can be useful to positively influence microbial community function such as selectively enhancing desirable active microbial populations to benefit health of the gut and the animal.

Lythrum salicaria Ellagitannins Stimulate IPEC-J2 Cells Monolayer Formation and Inhibit Enteropathogenic Escherichia coli Growth and Adhesion.

Lythrum salicaria herb (LSH) was applied in diarrhea therapy since ancient times. Despite empirically referenced therapeutic effects, the bioactivity mechanisms and chemical constituents responsible for pharmacological activity remain not fully resolved. Taking into consideration the historical use of LSH in treatment of diarrhea in humans and farm animals, the aim of the study was to examine in vitro the influence of LSH and its C-glycosylic ellagitannins on processes associated with maintaining intestinal epithelium integrity and enteropathogenic Escherichia coli (EPEC) growth and adhesion. LSH was not only inhibiting EPEC growth in a concentration dependent manner but also its adhesion to IPEC-J2 intestinal epithelial cell monolayers. Inhibitory activity toward EPEC growth was additionally confirmed ex vivo in distal colon samples of postweaning piglets. LSH and its dominating C-glycosylic ellagitannins, castalagin (1), vescalagin (2), and salicarinins A (3) and B (4) were stimulating IPEC-J2 monolayer formation by enhancing claudin 4 production. Parallelly tested gut microbiota metabolites of LSH ellagitannins, urolithin C (5), urolithin A (6), and its glucuronides (7) were inactive. The activities of LSH and the isolated ellagitannins support its purported antidiarrheal properties and indicate potential mechanisms responsible for its beneficial influence on the intestinal epithelium.

Effects of dietary L-arginine supplementation to early pregnant mares on conceptus diameter-Preliminary findings.

The importance of the amino acid L-arginine (ARG) for conceptus growth and litter size has been demonstrated in various species. L-arginine is part of embryo-derived polyamines, a substrate for nitric oxide synthase and stimulates protein synthesis by the embryo. In the present study, we have investigated whether dietary L-arginine supplementation stimulates early conceptus growth in mares. Warmblood mares with singleton pregnancies received either an arginine-supplemented diet (approximately 0.0125% of body weight, n = 12) or a control diet (n = 11) from days 15 to 45 after ovulation. Diameter of the embryonic vesicle (days 14, 17, 20 of pregnancy) and size of the embryo respective foetus (length and maximal diameter, days 25-45 of pregnancy at 5-day intervals) were determined by transrectal ultrasound. At foaling, weight and size of the foal and the placenta were determined. Blood for determination of equine chorionic gonadotrophin (eCG) and progestin concentrations was collected repeatedly. Neither eCG nor progestin concentration in plasma of mares differed between groups at any time. No effects of arginine treatment on diameter of the embryonic vesicle between days 14 and 20 of pregnancy were detected. Diameter of the embryo/foetus on days 40 to 45 of pregnancy strongly tended to be enhanced by arginine supplementation (p = 0.06). Weight and size of neither the foal nor placenta at birth differed between groups. In conclusion, L-arginine supplementation was without negative effects on early equine embryos and may support embryonic growth at the beginning of placentation.