In vitro fermentability and physicochemical properties of fibre substrates and their effect on bacteriological and morphological characteristics of the gastrointestinal tract of newly weaned piglets

Abstract

Fermentability of fibre has a great impact on the bacterial flora along the gastrointestinal tract of newly weaned piglets. Therefore, this parameter was determined by incubating in vitro different fibre substrates (chicory roots, sugar beet pulp, wheat bran and corn cobs) with contents of jejunum or caecum sampled from slaughtered pigs. Incubating with small intestinal contents, lactic acid was the only fermentation product. Fermentability was highest for chicory roots, followed by wheat bran and sugar beet pulp, while corn cobs were not fermented. Based on SCFA formed in the incubations with caecal contents, ranking of the fermentability of the fibre substrates was in the same order. The effect of adding different fibre substrates to diets of newly weaned piglets on bacteriological and morphological aspects of the gastrointestinal tract was also investigated. In Experiment 1 three groups of five piglets, weaned at four weeks of age, received a control feed (C), C supplemented with corn cobs (50 g/kg) or with chicory roots (20 g/kg). In Experiment 2, diet C was supplemented with sugar beet pulp (120 g/kg) or with wheat bran (75 g/kg). After three weeks animals were euthanized and digesta were sampled from stomach, proximal and distal jejunum, caecum and colon. Furthermore, mucosal scrapings were prepared and tissue samples were taken from jejunum, caecum and colon. Viscosity was determined for jejunal, caecal and colon contents. Corn cobs in the feed increased the number of total bacteria, lactobacilli and bifidobacteria in the stomach and proximal duodenum, while a decreased count of streptococci in distal jejunum contents was noted. Chicory roots increased the counts of Escherichia coli in the distal jejunum and on the mucosa, while sugar beet pulp decreased the number of lactobacilli on the mucosa only. Wheat bran seemed to increase the count of E. coli in jejunal digesta and on the mucosa, and also the number of lactobacilli in the stomach and jejunum. Bifidobacterial numbers were increased but only in the proximal part of the jejunum. Fibre substrates affected the concentration of lactate and SCFA in different parts of the intestinal tract. Feeding corn cobs increased villus length in the proximal jejunum by 13%. The number of intra-epithelial lymphocytes in the villous epithelium of proximal and distal jejunum was decreased by corn cobs and chicory roots supplementation while beet pulp and wheat bran had the opposite effect. In Experiment 1, apoptotic index of the mucosa of the distal jejunum was very low and decreased when corn cobs were fed. Mitotic index in the crypts was only affected by the wheat bran diet and a small decrease was noted. It was concluded that the fermentability of fibre was not an ideal criterion for predicting its effects on the flora. The effect of fibres on viscosity of digesta was negligible probably explaining the lack of clear and consistent influences on the intestinal mucosa.     

Incorporation of galactomannans in the diet of newly weaned piglets: effect on bacteriological and some morphological characteristics of the small intestine

In search of substances replacing antibiotics as growth promoters for farm animals, non-digestible oligosaccharides (NDO) or non-starch polysaccharides (NSP) have been proposed as possible alternatives. In this context, the influence of galactomannans on bacteriological and morphological aspects of the gastrointestinal tract in weanling pigs was investigated. Four groups of five newly weaned piglets received one of the following diets: control feed (C), C supplemented with guar gum (1%), C supplemented with locust bean gum (1%) and C supplemented with 10% of carob tree seeds meal as source of locust bean gum. The animals were euthanized after 11-12 days and digesta were sampled in stomach, jejunum (proximal and distal) and caecum, while mucosal scrapings and ring shaped tissue samples were taken of proximal and distal jejunum. On these samples bacteriological, biochemical and morphological determinations were carried out. Total count of bacteria in digesta and mucosal scrapings was not influenced by the different diets, with the exception of the proximal jejunum where a small decrease (0.5 log10 CFU) was noted with the guar gum and carob tree seeds diet. The number of E. coli increased by feeding both gums and carob tree seeds. With the latter diet, higher counts of streptococci were observed. In agreement with the lower concentration of lactic acid in jejunal contents, guar gum decreased the number of lactobacilli. Locust bean gum decreased the molar proportion of acetate in caecal contents while butyrate and valerate were augmented. Feeding the carob tree seeds resulted in shorter villi and a lower villus height/crypt depth ratio in the jejunum mucosa, which was an indication for a faster renewal rate of the epithelium. Both locust bean gum feeds significantly lowered the mitotic index in the crypts of the small intestine. Only with the carob tree seeds diet, viscosity of jejunal contents was increased. In conclusion, the effects of the addition of 1% of pure guar gum or locust bean gum were inconsistent and not very outspoken, whereas 10% of carob tree seeds meal in the diet resulted in influences on intestinal characteristics at the bacteriological and morphological level.     

Effect of inulin supplementation on selected gastric, duodenal, and caecal microbiota and short chain fatty acid pattern in growing piglets

We explored whether bifidobacteria and lactobacilli numbers and other selected bacteria in the upper intestine and the caecum of growing pigs were affected by diet and intake of inulin. Starting at two weeks after weaning (28 d) 72 pigs were fed two types of diets (wheat/barley (WB) or maize/gluten (MG)), without or with 3% inulin (WB + I, MG + I) for three and six weeks. Intestinal bacteria were quantified by fluorescence-in-situ-hybridization (n = 8/group). Duration of feeding had no effect on the variables tested, so data for both periods were pooled. Gastric total bacteria amounted to log(10) 7.4/g digesta. Bifidobacteria were detected in stomach and duodenum two weeks after weaning and disappeared thereafter. In jejunum and caecum bifidobacteria were present at a level of log(10) 7.0/g digesta. Inulin did not alter numbers of lactobacilli, bifidobacteria, enterococci, enterobacteria and bacteria of the Clostridium coccoides/Eubacterium rectale-group. Inulin disappearance in stomach plus jejunum was higher with the MG diet (73.7 vs. 60.7%, p = 0.013). Caecal acetate was lower in inulin-supplemented diets (p < 0.05) whereas propionate and butyrate were higher in pigs fed the WB diets (p < 0.05). With the WB diet total caecal short chain fatty acids concentration was higher which resulted in a lower pH value (p < 0.05).     

Incorporation of galactomannans in the diet of newly weaned piglets: Effect on bacteriological and some morphological characteristics of the small intestine

In search of substances replacing antibiotics as growth promoters for farm animals, non-digestible oligosaccharides (NDO) or non-starch polysaccharides (NSP) have been proposed as possible alternatives. In this context, the influence of galactomannans on bacteriological and morphological aspects of the gastrointestinal tract in weanling pigs was investigated. Four groups of five newly weaned piglets received one of the following diets: control feed (C), C supplemented with guar gum (1%), C supplemented with locust bean gum (1%) and C supplemented with 10% of carob tree seeds meal as source of locust bean gum. The animals were euthanized after 11?–?12 days and digesta were sampled in stomach, jejunum (proximal and distal) and caecum, while mucosal scrapings and ring shaped tissue samples were taken of proximal and distal jejunum. On these samples bacteriological, biochemical and morphological determinations were carried out. Total count of bacteria in digesta and mucosal scrapings was not influenced by the different diets, with the exception of the proximal jejunum where a small decrease (0.5 log₁₀ CFU) was noted with the guar gum and carob tree seeds diet. The number of E. coli increased by feeding both gums and carob tree seeds. With the latter diet, higher counts of streptococci were observed. In agreement with the lower concentration of lactic acid in jejunal contents, guar gum decreased the number of lactobacilli. Locust bean gum decreased the molar proportion of acetate in caecal contents while butyrate and valerate were augmented. Feeding the carob tree seeds resulted in shorter villi and a lower villus height/crypt depth ratio in the jejunum mucosa, which was an indication for a faster renewal rate of the epithelium. Both locust bean gum feeds significantly lowered the mitotic index in the crypts of the small intestine. Only with the carob tree seeds diet, viscosity of jejunal contents was increased. In conclusion, the effects of the addition of 1% of pure guar gum or locust bean gum were inconsistent and not very outspoken, whereas 10% of carob tree seeds meal in the diet resulted in influences on intestinal characteristics at the bacteriological and morphological level.     

Production,partial characterization and use of fungal cellulase-free xylanases in pulp bleaching

Seven fungal strains were screened for their ability to produce cellulase-free xylanases that could be used in pretreatment of sulphite pulp prior to bleaching. The potential xylanase producers were subjected to shake flask fermentations using four different carbon sources: wheat bran, corn cobs, oat spelts xylan and bleach plant effluent. When grown on corn cobs, Aspergillus foetidus (ATCC 14916) produced significant levels of xylanase (547.4 U/ml), accompanied however by 6.6 U/ml of cellulase activity. Two other strains, Aspergillus oryzae (NRRL 1808) and Gliocladium viride (CBS 658.70), produced high yields of cellulase-free xylanase on oat spelts xylan. The crude enzymes of these two isolates were characterized with respect to pH and temperature optima and stability in order to standardize the optimum conditions for their use on pulp. Although the two xylanases differed in their abilities to remove reducing sugars from pulp, their biobleaching abilities, when assessed in hydrogen peroxide delignification of pulp, were very similar: both of them increased brightness by 1.4 points and removed 7% of hemicellulose from pulp.     

Feed ingredients differing in fermentable fibre and indigestible protein content affect fermentation metabolites and faecal nitrogen excretion in growing pigs

To study the fermentation characteristics of different non-conventional dietary fibre (DF) sources with varying levels of indigestible CP content and their effects on the production of fermentation metabolites and on faecal nitrogen (N) excretion, an experiment was conducted with 40 growing pigs (initial BW 23 kg) using wheat bran (WB), pea hulls (PH), pea inner fibres (PIF), sugar beet pulp (SBP) or corn distillers dried grains with solubles (DDGS). The diets also contained soya protein isolate, pea starch and sucrose, and were supplemented with vitamin-mineral premix. Faecal samples were collected for 3 consecutive days from day 10, fed with added indigestible marker (chromic oxide) for 3 days from day 13 and pigs were slaughtered on day 16 from the beginning of the experiment. Digesta from the ileum and colon were collected and analysed for short-chain fatty acids (SCFA) and ammonia (NH3) content. The apparent total tract N digestibility was the lowest (P < 0.001) in diets based on DDGS (74%), medium in diets with WB and SBP (76% each) and highest in those with PIF and PH (79% and 81%, respectively). Expressed per kg fermented non-starch polysaccharides (NSP), faecal N excretion was higher with DDGS and WB diets (130 and 113 g/kg NSP fermented, respectively) and lower with PIF, PH and SBP diets (42, 52 and 55 g/kg NSP fermented, respectively). The PH-based diets had the highest (P < 0.05) SCFA concentrations, both in the ileum and the colon (27 and 122 mMol/kg digesta, respectively). The highest NH3 concentration was also found in the colon of pigs fed with PH (132 mMol/kg digesta). Loading plot of principle component analysis revealed that the CP : NSP ratio was positively related with faecal N excretion and NH3 concentration in colon contents, whereas negatively related with SCFA concentration in colon contents. In conclusion, pea fibres and SBP increased SCFA and reduced NH3 concentration in the pig's intestine and reduced faecal N excretion, which makes pea fibres and SBP an interesting ingredient to use in pig diet to improve the positive effect of DF fermentation on the gastrointestinal tract and reduce faecal N excretion.     

Release of ferulic acid from agroindustrial by-products by the cell wall-degrading enzymes produced by Aspergillus niger I-1472

Aspergillus niger I-1472 was grown on sugar beet pulp to produce cell wall polysaccharide-degrading enzymes, including feruloyl esterases. Compared to enzymatic activities measured in commercially available mixtures previously used for the release of ferulic acid, the A. niger enzymes were more various. These enzymes were tested to release ferulic acid from sugar beet pulp, maize bran, or autoclaved maize bran. They were as efficient as the commercial mixture to release ferulic acid from sugar beet pulp. On the other hand, they were much more efficient to release ferulic acid from maize bran after autoclaving pretreatment, as 95% of ferulic acid ester were solubilized. Thus, A. niger enzymes exhibited a high interest in the release of ferulic acid from various agro-industrial by-products.     

Effects of source of fibre on the development and pH of the gastrointestinal tract of broilers

The effects of different sources of dietary fibre on gastrointestinal tract (GIT) development and digesta pH were studied in 25-day-old broilers. There were four diets: a control diet low in fibre content (37.3 g neutral detergent fibre and 16.0 g acid detergent fibre/kg) and three additional diets in which 30 g/kg of cellulose (CEL), sugar beet pulp (SBP) or oat hulls (OH) were included at the expenses of sepiolite (a complex magnesium silicate clay). The relative weight (RW, %BW) of the GIT was greater with the SBP – than with the CEL – and the control diets. Also, gizzard RW was higher with the diet containing OH than with the diet containing CEL or the control diets, with the SBP diet being intermediate (P≤0.001). Fibre inclusion modified the pH of the GIT (P≤0.05) but the effects depended on the type of fibre. Crop pH was higher and proventriculus pH lower with SBP than with the other diets. However, gizzard pH was reduced with SBP and OH but not with CEL (P≤0.05). The pH of the digesta of the duodenum was similar for all diets but that of the jejunum, Meckel's diverticulum and ileum increased with SBP and decreased with CEL inclusion (P≤0.05). We concluded that dietary fibre affects the development and pH value of the different segments of GIT and that the effects differ according to the physico-chemical characteristics of the source of fibre used.     

Effect of insoluble fibre on intestinal morphology and mRNA expression pattern of inflammatory,cell cycle and growth marker genes in a piglet model

The effects of insoluble dietary fibre differing in lignin content on intestinal morphology and mRNA expression was tested in an animal model of 48 weaned piglets. Engaged fibre sources were wheat bran (rich in cellulose and hemicellulose) and pollen from Chinese Masson pine (Pinus massoniana) (rich in lignin), respectively. The fibre sources were added to a basal diet as follows: no addition (control), 3.0% wheat bran, 1.27% pine pollen, and 2.55% pine pollen. The 12 animals of each feeding group were fed four experimental diets ad libitum for 37 days and were then slaughtered for retrieving tissue samples from stomach, jejunum, ileum, colon and mesenterial lymph nodes. Both fibre sources increased villus height of mucosa in jejunum (+10% on average) and ileum (+16% on average). Results of mRNA expression rates of inflammatory, cell cycle and growth marker genes (NFκB, TNFα, TGFβ, Caspase3, CDK4, IGF1) were specific to fibre source and tissue: wheat bran induced an up-regulation of NFκB in stomach and jejunum, as well as TNFα and TGFβ, and Caspase3 in jejunum. Pine pollen induced down regulation of NFκB, TNFα, TGFβ, Caspase3, CDK4 and IGF1 in the colon as well as up-regulation of NFκB and TGFβ in mesenterial lymph nodes. Finally, an overall data comparison based on a hierarchical cluster analysis showed a close relation between gene regulation in different gut sections and organs, as well as between small intestine morphology and zootechnical performance.     

Influence of different fibre sources on digestibility and nitrogen and energy balances in growing pigs

The present study was undertaken to investigate how three different fibre sources, sugar beet pulp, soya bean hulls and pectin residue, constituting 15% of diets for growing pigs, influenced daily body gain, feed conversion, apparent faecal digestibility and nitrogen and energy balances. Eight castrated crossbreed pigs (30-80 kg live weight) were used in a replicated 4 x 4 Latin-square design with one control diet and three fibre containing diets. Daily body weight gain and feed conversion were not affected by the dietary treatments. The apparent faecal digestibility of organic matter (OM) and energy were significantly lower for the fibre diets (OM: 0.81-0.85; energy: 0.78-0.83) compared to the control diet (OM: 0.88; energy: 0.86). The apparent faecal digestibility of crude protein (CP) was lower for the fibre diets (0.71-0.78) compared to the control diet (0.83), although it was only significantly lower for the sugar beet pulp and pectin residue diets. The pectin residue diet, which contained the highest amount of dietary fibre, lignin and insoluble non-starch polysaccharides, had the lowest digestibility of OM, CP and energy. There was a tendency (p = 0.07) for a diet effect on retained nitrogen in proportion to digested nitrogen, where the sugar beet pulp and pectin residue diets had numerically the highest values. Heat production and retained energy in proportion to metabolizable energy intake were not affected by fibre inclusion. It was concluded that the inclusion of sugar beet pulp, soya bean hulls and pectin residue in diets for growing pigs decreased the apparent faecal digestibility and in the diets with sugar beet pulp and pectin residue higher utilization of digested nitrogen for retention compensated for the lower amount of digested nitrogen.     

Aspergillus niger I-1472 and Pycnoporus cinnabarinus MUCL39533, selected for the biotransformation of ferulic acid to vanillin, are also able to produce cell wall polysaccharide-degrading enzymes and feruloyl esterases

The filamentous fungal strains Aspergillus niger I-1472 and Pycnoporus cinnabarinus MUCL39533, previously selected for the bioconversion of ferulic acid to vanillic acid and vanillin respectively, were grown on sugar beet pulp. A large spectrum of polysaccharide-degrading enzymes was produced by A. niger and very few levels of feruloyl esterases were found. In contrast, P. cinnabarinus culture filtrate contained low amount of polysaccharide-degrading enzymes and no feruloyl esterases. In order to enhance feruloyl esterases in A. niger cultures, feruloylated oligosaccharide-rich fractions were prepared from sugar beet pulp or cereal bran and used as carbon sources. Number of polysaccharide-degrading enzymes were induced. Feruloyl esterases were much higher in maize bran-based medium than in sugar beet pulp-based medium, demonstrating the ability of carbon sources originating from maize to induce the synthesis of feruloyl esterases. Thus, A. niger I-1472 could be interesting to release ferulic acid from sugar beet pulp or maize bran.     

Fermented and extruded wheat bran in piglet diets: impact on performance,intestinal morphology,microbial metabolites in chyme and blood lipid radicals

The aim of the present study was to evaluate the influence of native, fermented and extruded wheat bran on the performance and intestinal morphology of piglets. Additionally, short-chain fatty acids (SCFA), biogenic amines, ammonia, lactic acid, pH as well as E. coli and lactic acid bacterial counts were analysed in digesta samples from three gut sections. Furthermore, the antioxidant potential in blood samples was evaluated based on the lipid radicals formed. For this purpose, 48 newly weaned piglets (28 d old) were allocated to one of the four different dietary treatment groups: no wheat bran (Control), native wheat bran, fermented wheat bran as well as extruded wheat bran. Wheat bran variants were included at 150 g/kg into the diets. All diets were mixed to reach the calculated isonitrogenic nutrient contents. Gut tissue and digesta samples were collected from the proximal jejunum, the terminal ileum and the colon ascendens, blood samples directly at slaughter. Although none of the dietary interventions had an impact on performance parameters, the amount of goblet cells in the ileum was increased upon feeding native and extruded wheat bran, compared to fermented bran (p < 0.05). The E. coli counts in colonic chyme were significantly lower (p < 0.05) in the Control group compared to the groups fed with wheat bran. The concentration of SCFA showed differences for minor compounds (p < 0.05), while linear contrast analyses revealed a reduced concentration of total SCFA in the colon following the feeding of modified wheat bran compared to native wheat bran. This may suggest that several compounds are more easily digested already in the ileum, resulting in a reduced nutrient flow into the large intestine and therefore less unexploited digesta is available as substrate for the microorganisms there. Fermentation also resulted in a significant decrease of methylamine in the colon (p < 0.05), while other biogenic amines in the ileum and colon showed no statistically significant differences. The formation of lipid radicals was decreased (p < 0.05) after feeding native wheat bran compared to the Control group. These results suggest that fermentation and extrusion of wheat bran exert some different impact regarding their physiological mode of action.     

Influence of different fibre sources on digestibility and nitrogen and energy balances in growing pigs

Abstract

The present study was undertaken to investigate how three different fibre sources, sugar beet pulp, soya bean hulls and pectin residue, constituting 15% of diets for growing pigs, influenced daily body gain, feed conversion, apparent faecal digestibility and nitrogen and energy balances. Eight castrated crossbreed pigs (30 – 80 kg live weight) were used in a replicated 4 × 4 Latin-square design with one control diet and three fibre containing diets. Daily body weight gain and feed conversion were not affected by the dietary treatments. The apparent faecal digestibility of organic matter (OM) and energy were significantly lower for the fibre diets (OM: 0.81 – 0.85; energy: 0.78 – 0.83) compared to the control diet (OM: 0.88; energy: 0.86). The apparent faecal digestibility of crude protein (CP) was lower for the fibre diets (0.71 – 0.78) compared to the control diet (0.83), although it was only significantly lower for the sugar beet pulp and pectin residue diets. The pectin residue diet, which contained the highest amount of dietary fibre, lignin and insoluble non-starch polysaccharides, had the lowest digestibility of OM, CP and energy. There was a tendency (p = 0.07) for a diet effect on retained nitrogen in proportion to digested nitrogen, where the sugar beet pulp and pectin residue diets had numerically the highest values. Heat production and retained energy in proportion to metabolizable energy intake were not affected by fibre inclusion. It was concluded that the inclusion of sugar beet pulp, soya bean hulls and pectin residue in diets for growing pigs decreased the apparent faecal digestibility and in the diets with sugar beet pulp and pectin residue higher utilization of digested nitrogen for retention compensated for the lower amount of digested nitrogen.     

Biocontrol of Damping-off Diseases Caused by Rhizoctonia solani and Pythium ultimum with Alginate Prills of Gliocladium virens, Trichoderma hamatum and Various Food Bases

Alginate prills were formulated with the biomass of isolates of Gliocladium virens and Trichoderma spp. and various food bases (wheat bran, corn cobs, peanut hulls, soy fiber, castor pomace, cocoa hulls and chitin). Alginate prills with G. virens (Gl-21) biomass and all food bases except cocoa hull meal significantly reduced the damping-off of zinnia in a soil-less mix caused by Rhizoctonia solani and Pythium ultimum. The prills with bran, soy fiber, castor pomace or chitin resulted in stands similar to those in the non-infested control. In soil, prills with all the food bases and Thrichoderma hamatum (TRI-4) biomass controlled the damping-off of cotton caused by R. solani and gave stands comparable to, or better than, those in the non-infested control soil. Prills with all the food bases resulted in a proliferation of Gl-21 in a soil-less mix and of Gl-21 and TRI-4 in soil. Prills with food bases and TRI-4 biomass reduced the survival of R. solani in infested beet seed to less than 30%, with bran and chitin being the most effective food bases; prills with Gl-21 biomass and all food bases also reduced the survival of R. solani in beet seed, but not as much as did prills with TRI-4 biomass. In prills containing wheat bran, soy fiber or chitin, the biocontrol isolate Th-58 (T. harzianum) was almost as effective as TRI-4, but isolate Gl-3 (G. virens) was less effective. There was no significant interaction between the biocontrol fungus and the food base. The results suggest that the intrinsic properties of a selected fungus isolate are more important than some formulation variables in biocontrol.     

Effects of fibre-degrading enzymes in combination with different fibre sources on ileal and total tract nutrient digestibility and fermentation products in pigs

ABSTRACT

The study was conducted to determine effects of a complex of fibre-degrading enzymes (xylanase, cellulase and β-glucanase) on nutrient digestibility, fibre fermentation and concentrations of short chain fatty acids (SCFA) at different parts of digestive tract in pigs fed different fibre-rich ingredients. A total of 36 barrows fitted with T-cannulas in the distal ileum (initial body weight of 41.1 ± 2.7 kg) were randomly allotted to six dietary treatments with three different high-fibre diets including maize bran (MB), sugar beet pulp (SBP) and soybean hulls (SH) with or without supplementation of fibre-degrading enzymes. Enzyme supplementation improved (p < 0.05) apparent ileal digestibility (AID) of dietary gross energy (GE), crude protein, dry matter (DM), organic matter (OM), total dietary fibre (TDF), neutral detergent fibre (NDF) and apparent total tract digestibility (ATTD) of dietary GE, DM, OM, TDF, insoluble dietary fibre (IDF) when pigs were fed MB, SBP or SH diets. When compared to the SBP and SH diets, the AID of GE, DM, ash, OM and NDF in diet MB was higher (p < 0.05), but the hindgut disappearance and ATTD of nutrients, except for ether extract and crude ash, were lower (p < 0.05). Enzyme supplementation increased acetate and total SCFA concentrations in ileal digesta and faeces of pigs. In conclusion, enzyme addition improved IDF fermentation and SCFA concentration in the whole intestine of pigs, and there was a large variation of digestibility of fibre components among MB, SH and SBP owing to their different fibre composition. Therefore, fibre-degrading enzymes should be applied to fibrous diets to improve efficient production of swine, especially considering low fibre digestibility of fibre-rich ingredients.     

Optimization of nutrient medium containing agricultural wastes for xylanase production by Aspergillus niger B03 using optimal composite experimental design

The xylanase biosynthesis is induced by its substrate - xylan. The high xylan content in some of the wastes like corn cobs and wheat bran makes them an accessible and cheap source of inducers. Nutrient medium for xylanase biosynthesis in submerged cultivation of Aspergillus niger B03 has been optimized. The optimization process was analyzed using optimal composite experimental design and response surface methodology. The predicted by the regression model optimum components of nutrient medium are as follows (g/l): (NH(4))(2)HPO(4) 2.6, urea 0.9, corn cobs 24.0, wheat bran 14.6 and malt sprout 6.0. Five parallel experiments have been carried out, at definite, optimum components concentrations of the nutrient medium, and a mean value of the activity Y=996.30 U/ml has been obtained. The xylanase activity, obtained with the optimized nutrient medium is 33% higher than the activity, achieved with the basic medium.     

Production and characterization of xylanases of a Bacillus strain isolated from soil

Xylanase production was performed by growing a Bacillus isolate on agricultural by-products, wheat straw, wheat bran, corn cobs and cotton bagasse. A maximum xylanase activity of 180 U/ml was obtained together with a cellulase activity of 0.03 U/ml on 4 (w/v) corn cobs. Electrophoretic analysis showed the presence of three endo--1, 4-xylanases having molecular weights of about 22, 23 and 40 kDa. Xylanolytic activity was stable up to 50 °C in the pH range of 4.5–10 and the highest activity was observed at 70 °C and pH 6.5.     

Production of feruloyl/ρ-coumaroyl esterase activity by Penicillium expansum, Penicillium brevicompactum and Aspergillus niger

Extracellular esterase production by Penicillium expansum, Penicillium brevicompactum and Aspergillus niger was determined in both liquid and solid-state culture. Methyl ferulate was used as the main carbon source in liquid culture whereas wheat bran and sugar beet pulp were used in solid-state culture. Extracted enzyme for each fungus showed activity in the presence of ONP butyrate, methyl ferulate, methyl coumarate and two 'natural'feruloylated carbohydrate esters. Higher enzyme recoveries were obtained using wheat bran in solid-state culture. Higher levels of feruloyl esterase activity were recovered from P. expansum on all feruloylated substrates than from P. brevicompactum or A. niger. Using ONP butyrate as substrate the pH and temperature optima for the esterases of both Penicillium spp. were 6.0 and 25–30°C. Aspergillus niger esterase activity showed a broader temperature range with an optimum at 40°C.     

Different fibrous ingredients and coarsely ground maize affect hindgut fermentation in the pig in vitro but not Salmonella Typhimurium survival

The fermentation kinetics of both fibre choice and maize particle size were studied in vitro from the ileal contents of cannulated pigs given five different experimental diets. Additionally, in vitro batch systems were used to study the quantitative effect of fibre choice and maize particle size on Salmonella Typhimurium growth. Freeze-dried ileal effluents obtained from five cannulated pigs given the five experimental diets in a latin square were used as incubation substrates. The experimental diets consisted of: (1) a standard maize-based diet (ST) where all the ingredients were milled to pass through a 2.5-ml screen, (2) the same diet with the maize ground using a 4.0-ml screen (CG), (3 and 4) two diets in which the maize was partially replaced by (3) sugar beet pulp (80 g/kg; BP) or (4) wheat bran (100 g/kg; WB), or (5) a combination of 80 g sugar beet pulp and 100 g wheat bran/kg (diet HF). Results showed that substrate from BP and HF generated more gas than substrate from ST (P<0.001). In addition, short chain fatty acids production during the entire incubation period was higher (P=0.001) for all the substrates compared to ST substrate (P<0.001) where lower acetate and higher propionate, valerate and branched-chain fatty acids molar ratios were measured. The molar ratio of acetate was highest and the branched-chain fatty acids was lowest for substrates from diets that included sugar beet pulp. The butyrate molar ratio was lower for all substrates compared to the ST diet except for the WB diet. After 12 h of incubation, HF substrate presented the lowest ammonia (91.7 mg/l vs. 125.3 mg/l, P=0.014) and the highest purine bases concentration (0.28 μmol/ml vs. 0.22 μmol/ml, P=0.009). None of the substrates showed any inhibitory effect on Salmonella growth when the batch systems were inoculated with S. Typhimurium. In summary, under the in vitro conditions used, the changes in hindgut fermentation promoted by the inclusion of different fibrous ingredients or by different maize particle size did not produce an inhibitory effect on the growth of S. Typhimurium.     

Feruloyl esterases as a tool for the release of phenolic compounds from agro-industrial by-products

Agro-industrial by-products are a potential source of added-value phenolic acids with promising applications in the food and pharmaceutical industries. Here two purified feruloyl esterases from Aspergillus niger, FAEA and FAEB were tested for their ability to release phenolic acids such as caffeic acid, p-coumaric acid and ferulic acid from coffee pulp, apple marc and wheat straw. Their hydrolysis activity was evaluated and compared with their action on maize bran and sugar beet pulp. The specificity of both enzymes against natural and synthetic substrates was evaluated; particular attention was paid to quinic esters and lignin monomers. The efficiency of both enzymes on model substrates was studied. We show the ability of these enzymes to hydrolyze quinic esters and ester linkages between phenolic acids and lignin monomer.