Branched chain amino acids as source of specific branched chain volatile fatty acids during the fermentation process of fish sauce

Summary.  The source of the formation of branched chain volatile fatty acids (VFA) in fish sauce was investigated. Certain branched VFA were derived from the degradation of specific amino acids as iso-butyric acid from valine and iso-valeric acid from leucine. Short and long straight chain VFA were significantly higher in the linoleic acid added sample than in the control but did not significantly bring changes to the branched chain VFA. It is suggested that straight chain VFA developed from fish fats. Alanine and isoleucine did not have a clear influence on the production of volatile fatty acids. Received November 23, 2001 Accepted June 20, 2002 Published online December 18, 2002 RID="*" ID="*" Part of this paper was presented in the 7th International Congress on Amino Acids and Proteins in Vienna, Austria from August 6–10, 2001. Authors' address: Norlita G. Sanceda, Ph.D., Institute of Environmental Science for Human Life, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112, Japan, Fax: + 81-3-5978-5805, E-mail: lita@cc.ocha.ac.jp     

Iso-alkane oxidation by aPseudomonas Part I.—Metabolism of 2-methylhexane

Summary Pseudomonas cells adapted to 2-methylhexane oxidize 5-methylhexanoic andiso-valeric acids rapidly. The oxidation rates of 2-methylhexanoic and propionic acids are appreciably lower. It can be concluded that primary attack on the C₆ atom is distinctly favoured over that on C₁. By use of gas-liquid chromatography, 2-methylhexanoic, 5-methylhexanoic andiso-valeric acids were shown to be formed when heptane-grown cells were incubated with 2-methylhexane. When 2-methylhexane-grown cells were used the amount of 5-methylhexanoic acid decreased compared with the amount of the 2-methyl isomer. Moreover,iso-valeric acid could not be detected. The results make it probable that degradation of 2-methylhexanevia C₆, comprising 5-methylhexanoic andiso-valeric acid, is accompanied by a second pathway via 2-methylhexanoic acid. The latter pathway is of minor importance, in particular in 2-methylhexane-adaptedPseudomonas cells.     

猪不同肠段微生物体外培养对蛋氨酸的代谢特性

【目的】旨在通过微生物体外发酵技术,以回肠微生物为参照,研究猪盲肠及结肠微生物对在小肠微生物中代谢率较低的蛋氨酸的代谢特性。【方法】采集4头健康100 kg左右杜×长×大杂交猪的盲肠、结肠与回肠食糜作为接种物,分别接种于10 mmol/L蛋氨酸的培养基中,37°C体外培养24 h。分别设含蛋基酸溶液和含各肠段食糜接种物的空白对照组。【结果】(1)不同肠段微生物以蛋氨酸为底物体外发酵,盲肠组蛋氨酸消失率(21.9%)显著高于结肠组(16.7%)与回肠组(16.3%)(P0.05)。盲肠组总SCFA量显著高于结肠与回肠组(P0.05),伴随着p H值下降程度最高;盲肠组MCP产量也显著高于结肠与回肠组(P0.05);在产气量与NH3-N浓度上,盲肠组与结肠组均显著低于回肠组(P0.05)。(2)以蛋氨酸为底物体外发酵,门水平上,总菌、厚壁菌门含量在各肠段组间无显著差异(P0.05),拟杆菌门含量在盲肠组最高;与不加蛋氨酸底物的对照组比较,三个肠段试验组总菌、厚壁菌门含量均显著高于对照组(P0.05),而拟杆菌门含量在试验组与对照组间差异不显著(P0.05)。属水平上,盲肠组和结肠组大肠杆菌属数量显著低于回肠组(P0.05),而柔嫩梭菌属和梭菌XIV属数量在盲肠组和结肠组均高于回肠组;各肠段组间双歧杆菌数量无显著差异(P0.05)。【结论】以蛋氨酸为底物,体外培养猪盲肠微生物对蛋氨酸代谢率高于回肠微生物,伴随着其他发酵参数的变化,并且发酵产生更多的菌体蛋白。相比于回肠微生物发酵,大肠微生物发酵后,柔嫩梭菌属和梭菌XIV属数量较高,而大肠杆菌属数量较低。     

Influence of particle size and pH on anaerobic degradation of cellulose by ruminal microbes

Batch experiments were performed to investigate the influence of cellulose particle size and pH on the anaerobic degradation of crystalline cellulose by ruminal microbes. At a particle size of 50 μm there was a higher hydrolysis and acidogenesis rate, and a reduced degradation time, than for 100-μm particles. Reduction in cellulose particle size resulted in decreased methane production, but an increase of soluble products. Cellulose degradation increased with pH from pH 6.0 to 7.5, whereas at pH⩽5.5 there was no degradation. The inhibitory effect of low pH (⩽5.5) on ruminal microbes was not completely remedied even when the pH of the medium was adjusted to a neutral range. In an anaerobic cellulosic waste degrading system inoculated with ruminal microbes the fermentation system should therefore be maintained above pH 6.0. In all cases, volatile fatty acids were the major water-soluble products of cellulose degradation; acetate and propionate accounted for more than 90% of the volatile fatty acid total.     

Continuous feeding of antimicrobial growth promoters to commercial swine during the growing/finishing phase does not modify faecal community erythromycin resistance or community structure

Aims: To investigate the effect of continuous feeding of antimicrobial growth promoters (tylosin or virginiamycin) on the swine faecal community. Methods and Results: The study consisted of two separate on‐farm feeding trials. Swine were fed rations containing tylosin (44 or 88 mg kg^?1 of feed) or virginiamycin (11 or 22 mg kg^?1 of feed) continuously over the growing/finishing phases. The temporal impact of continuous antimicrobial feeding on the faecal community was assessed and compared to nondosed control animals through anaerobic cultivation, the analysis of community 16S rRNA gene libraries and faecal volatile fatty acid content. Feeding either antimicrobial had no detectable effect on the faecal community. Conclusions: Erythromycin methylase genes encoding resistance to the macrolide–lincosamide–streptogramin B (MLS_B) antimicrobials are present at a high level within the faecal community of intensively raised swine. Continuous antimicrobial feeding over the entire growing/finishing phase had no effect on community erm‐methylase gene copy numbers or faecal community structure. Significance and Impact of the Study: Antimicrobial growth promoters are believed to function by altering gut bacterial communities. However, widespread MLS_B resistance within the faecal community of intensively raised swine likely negates any potential effects that these antimicrobials might have on altering the faecal community. These findings suggest that if AGP‐mediated alterations to gut communities are an important mechanism for growth promotion, it is unlikely that these would be associated with the colonic community.     

Effects of nondigestible oligosaccharides on Salmonella enterica serovar Typhimurium and nonpathogenic Escherichia coli in the pig small intestine in vitro

An in vitro intestinal tissue model was developed for the investigation of bacterial association in the pig small intestine under different dietary regimes. In preliminary experiments, jejunal and ileal tissue was taken from Danish Landrace pigs fed standard diet and inoculated with either Salmonella or nonpathogenic Escherichia coli strains. Higher numbers of salmonellae associated with the ileal tissues, but the numbers did not reach significance. Hence, jejunal sections were inoculated with nonpathogenic E. coli and ileal sections were inoculated with salmonellae in the presence of mannose or commercial nondigestible oligosaccharides (NDO) at 2.5%. There was a significant decrease in E. coli associated with the jejunum in the presence of mannose (P < 0.05). Furthermore, in pigs fed a diet supplemented with commercial NDO at 4% there was a significant reduction in the numbers of E. coli in jejunal organ cultures of pigs fed the FOS diet (P < 0.05). There was a reduction, though not a significant one, in the association of Salmonella sp. to the ileal sections of pigs fed the commercial FOS diet. The feeding of commercial GOS or its addition to organ cultures did not affect E. coli or Salmonella numbers.     

Reliable identification of Prevotella and Butyrivibrio spp. from rumen by fatty acid methyl ester profiles

Data for bacterial identification were provided by culturing anaerobic bacteria under standardized conditions followed by extraction and methylation of cellular long-chain fatty acids and gas chromatographic analysis. The databases of fatty acid methyl ester (FAMEs) profiles for two predominant ruminal genera,Prevotella andButyrivibrio, were created. Major long-chain cellular fatty acids found in the 23 analyzedPrevotella strains were 15:0 (anteiso), 15:0, 15:0 (iso) and 16:0. The strains ofPrevotella could be well identified on species level by the characteristic ratios among major fatty acids and by acids unique fatty for each species. The 45Butyrivibrio strains were grouped into 4 major and 2 minor groups according to FAMEs profiles. The major fatty acids for the bulk of theButyrivibrio strains were 14:0, 15:1, 16:0 and 16:0 (iso). This groups corresponded to those based on 16S rDNA sequences.     

Impact of diet composition on ileal digestibility and small intestinal morphology in early-weaned pigs fitted with a T-cannula

Piglets, separated from their dam at 12 days of age and fed a milk substitute hourly, were used as a model for suckling. Animals were fitted with a terminal ileal T-cannula and a jugular vein catheter. At 28 days of age, half of the pigs had a dietary change to a cereal-based weaner diet fed as slurry, and the others remained on milk substitute. Animals were labelled by oral administration of 15N-labelled yeast for 10 days (days 15 to 25). Blood samples were taken twice a day to monitor 15N enrichment of the blood plasma. Diets included polyethylenglycol (PEG 4000) to allow calculation of apparent ileal digestibility of nitrogen and individual amino acids. Ileal bacterial nitrogen was calculated from D-alanine content of the digesta. Furthermore, small intestinal (SI) villus height and crypt depth were measured. Feed intake was increased by the dietary change. The total nitrogen flow was 3.2 ± 0.4 g/day and 5.9 ± 0.4 for the milk and weaner diet, respectively. Endogenous nitrogen flow at the terminal ileum was similar for both groups (milk diet 2.4 ± 0.4 v. weaner diet 2.2 ± 0.3 g/day), whereas the bacterial nitrogen content (0.08 ± 0.01 g/day milk diet v. 0.15 ± 0.01 g/day weaner diet, P < 0.01) and exogenous nitrogen flow (0.94 ± 0.16 g/day milk diet v. 3.29 ± 0.12 g/day weaner diet, P < 0.001) increased significantly in the weaner-diet group. The ileal apparent digestibility coefficient of protein was 0.81 ± 0.06 and 0.68 ± 0.01 for the milk replacer and the weaner diet, respectively. Morphology measurements made along the SI at 25%, 50% and 75% were similar between piglets fed milk replacer and those fed a cereal-based weaner diet. The only statistical effect (P < 0.01) of dietary change was an increase in crypt depth in the weaner-diet group. In conclusion, pigs, following a dietary change analogous to weaning, lack the capacity to fully digest a standard weaner diet. This may result in an increased nutrient content entering the large intestine and an altered microbiota. In the absence of a period of anorexia, often associated with traditional weaning, we saw no evidence of villous atrophy, but report here a significant crypt hyperplasia, especially at the 75% level, as a result of dietary change.     

Aus der Versuchsstation für Tierernährung,Perchtoldsdorf/Wien

This study aimed to compare the effects of organic (proteinate) and inorganic (sulphate) copper (Cu) and zinc (Zn) supplements, in presence or absence of a mannan oligosaccharide (MOS) supplement, on mineral solubility and digestibility in pigs. Twenty-eight barrows (25 ± 4 kg) assigned randomly to four treatment groups were fed a corn-wheat-soya bean meal diet with 10 mg/kg of Cu and 100 mg/kg of Zn supplied as organic or inorganic supplement, and supplemented or not with 0.1% MOS. After an adaptation period, total faeces and urine were collected for a period of 6–7 days. Pigs were then euthanatised and digesta from ileum and caecum were collected. Apparent digestibility was calculated in ileum and caecum using titanium dioxide. The organic mineral supplement improved total (faecal) digestibility and retained/ingested ratio of Cu (p < 0.05) while reducing apparent digestibility of Zn in the ileum (p < 0.05) without effect on total digestibility of Zn. Solubilities of Cu and Zn in liquid fraction of ileum and caecum were not affected by mineral sources. Although MOS supplement increased Cu solubility in the ileum (p < 0.05), it had no effect on digestibility of Zn and Cu in ileum, caecum and faeces, retained/ingested ratio of Zn and Cu, or pH and volatile fatty acid concentration in ileal and caecal digesta. In conclusion, organic mineral supplement improved total digestibility and retained/ingested ratio of Cu in pigs but this cannot be attributed to its solubility in ileal and caecal digesta. The MOS supplement did not interfere with digestibility or dietary utilisation of Zn and Cu in pigs fed above the Zn and Cu requirements.     

Microbiology and ration digestibility in the hindgut of the ovine.

Contents of the terminal ilea, ceca-proximal colons, and terminal recta were obtained from nine sheep, three of which were fed 100% orchardgrass hay, three of which were fed 60% cracked corn-40% orchardgrass hay, and three of which were fed 80% cracked corn-20% orchardgrass hay. The digestibility of dry matter in the ceca was greatest when the all-hay diet was fed, whereas the percentage of cellulose digestion in the ceca increased with increasing levels of concentrate. For all diets, the total volatile fatty acid concentrations were higher in the ceca than in the other two sites. The cecal pH levels decreased with increased corn levels in the diet. The total microbiol numbers per gram of ileal and cecal contents increased in response to feeding of concentrate; however, across all diets, the ileal counts were 8% or less of the cecal counts. In contrast, the cellulolytic microbial numbers in the ilea were 50% or more of those in the ceca and were highest with the all-hay diet. Both bacterial and end product concentrations in the ceca were equivalent to those occurring in rumina. A total of 16 cellulolytic cultures were isolated and characterized from ileal and cecal contents of animals fed all three diets. Seven gram-negative-rod-shaped organisms were identified as Butyrivibrio fibrisolvens, whose capacity to digest cellulose exceeded that of several rumen strains. The guanine-plus-cytosine content of the DNA of one strain of B. fibrisolvens was 38.8 mol%, compared with the only previously reported value for this species of 41.2 mol%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of cell wall degrading enzymes on carbohydrate fractions and metabolites in stomach and ileum of pigs fed wheat bran based diets

Pigs were fed diets containing 40% wheat bran incubated with a water.‐acetic acid mixture (control, C) and a cellulase (Cel‐i) or xylanase (Xyl‐i) preparation or with addition of the cellulase (Cel‐a) or xylanase (Xyl‐a) preparation immediately before feeding. Stomach and ileal samples were analysed for pH, osmolality, soluble saccharides, volatile fatty acids (VFA) and lactic acid. Incubation of wheat bran resulted in a small reduction of NDF and an increase in the amount of soluble starch, ß‐glucans and saccharides (glucose, xylose and arabinose), especially after incubation with the cellulase preparation. Two hours after feeding, significantly higher arabinose and xylose concentrations were present in the stomach for diets Cel‐i, Cel‐a and Xyl‐i. In the ileum xylose and arabinose concentrations were higher 2 to 4 and 6 to 8 hours after feeding the enzyme‐treated diets. In stomach and ileum there were no differences between the diets in pH, osmolality, VFA and lactic acid concentrations, but ileal VFA concentration from 4 h after feeding tended to be higher for diets Cel‐i and Xyl‐i. It can be concluded that the amount of soluble saccharides in stomach and small intestine and the ileal VFA concentration may be increased by cell wall degrading enzyme preparations.     

Effect of nisin on two cultures of rumen ciliates

The effect of nisin (in the form of Nisaplin) was determined using two species of rumen ciliate protozoa in vitro, on their co-culture bacterial population, and volatile fatty acid concentration. Nisaplin did not affect the in vitro growth of Entodinium caudatum at concentrations of 50-400 mg/L during short-term treatment (5 d). Long-term application (30 d) of Nisaplin (100 mg/L) significantly decreased growth of the Epidinium ecaudatum forma caudatum et ecaudatum but not growth of E. caudatum. Nisaplin moderately supported the growth of E. caudatum after omission of wheat gluten (source of amino acids for protozoan growth). An inhibition of Gram-positive facultative anaerobic bacterial population in the protozoan cultures (lactobacilli, enterococci, staphylococci and amylolytic streptococci) was observed during long-term Nisaplin treatment. The concentration of volatile fatty acids significantly increased during the long-term Nisaplin treatment of both cultures. The propionate concentration in the mixture of volatile fatty acids was nearly twice higher on the account of the decreased concentration (from 74 to 63%) of acetate.     

Pathways of hydrocarbon dissimilation by aPseudomonas as revealed by chloramphenicol

Pseudomonas aeruginosa cells pre-grown on a particular hydrocarbon will oxidize other hydrocarbons as well. Degradation of these hydrocarbons proceeds to a point — depending on their structure — where new enzymes are needed for further degradation. Lack of these enzymes causes accumulation of products. However, secondary adaptation processes tend to decrease yields of intermediates, in particular when adaptation rates are high.By inhibiting these secondary adaptation processes with chloramphenicol (CAM), the amounts of various intermediates could be increased.Propionic acid was accumulated from heptane by hexane-grown cells in yields up to 60% (molar) calculated on heptane converted. The effect of CAM suggests that propionic acid is not subject to -oxidation (acrylate pathway) but is degraded via methylmalonate by adaptive enzymes.2-Methylhexane was converted for 30–40% (molar) intoiso-valeric acid by heptane-grown cells. A known pathway ofiso-valeric acid oxidation incorporates a carbon dioxide fixation step, and lack of this enzyme in heptane-grown cells probably causesiso-valeric acid accumulation.Heptene-1 incubation with heptane-grown cells resulted in a 30–40% conversion into 4-pentenoic plus 2, 4-pentadienoic acids. 6-Heptenoic acid was detected occasionally. A predominant attack at C₇ of the heptene-1 molecule is indicated at least for heptane-grown cells. Attack on the saturated end of the molecule seems well in line with the assumption that alkane oxidation by these bacteria is effected by oxygen transferring enzymes operating on a methyl group, as opposed to the action of a dehydrogenase and formation of an -olefin as the intermediate.     

Effects of Nondigestible Oligosaccharides on Salmonella enterica Serovar Typhimurium and Nonpathogenic Escherichia coli in the Pig Small Intestine In Vitro

An in vitro intestinal tissue model was developed for the investigation of bacterial association in the pig small intestine under different dietary regimes. In preliminary experiments, jejunal and ileal tissue was taken from Danish Landrace pigs fed standard diet and inoculated with either Salmonella or nonpathogenic Escherichia coli strains. Higher numbers of salmonellae associated with the ileal tissues, but the numbers did not reach significance. Hence, jejunal sections were inoculated with nonpathogenic E. coli and ileal sections were inoculated with salmonellae in the presence of mannose or commercial nondigestible oligosaccharides (NDO) at 2.5%. There was a significant decrease in E. coli associated with the jejunum in the presence of mannose (P < 0.05). Furthermore, in pigs fed a diet supplemented with commercial NDO at 4% there was a significant reduction in the numbers of E. coli in jejunal organ cultures of pigs fed the FOS diet (P < 0.05). There was a reduction, though not a significant one, in the association of Salmonella sp. to the ileal sections of pigs fed the commercial FOS diet. The feeding of commercial GOS or its addition to organ cultures did not affect E. coli or Salmonella numbers.     

Feeding level in the period previous to the late fattening phase influences fat composition at slaughter in free-ranged Iberian pigs

Abstract

A group of 11 pigs was fed with 70 g feed per kg of metabolic weight (H pigs) and another group of 11 pigs was fed with 50 g feed per kg of metabolic weight (L pigs). In both experimental groups (H and L pigs), it was observed that the higher initial proportion of C16:0, C18:0 and C18:2 (n-6) in backfat at the beginning of the free-range feeding period, the greater decrease rate of these fatty acid proportions regarding weight gain during the free-range fattening period took place. On the other hand, the greater initial proportion of C18:1 (n-9), the smaller increase rate in the concentration of this fatty acid was observed. The intramuscular neutral lipids from L pigs had higher C18:3 (n-3) and lower proportions of monounsaturated fatty acids (MUFA) than those from H pigs, while intramuscular polar lipids from L pigs had significantly higher proportions of C18:0, PUFA, C18:3 (n-3) and (n-3) and significantly lower MUFA and C18:1 (n-9) proportions than those from H pigs. The α-tocopherol concentration found in Longissimus dorsi from L pigs was significantly higher (p < 0.012) than those from H pigs.     

Enterobacterial and viral decay experimental models for anaerobic digestion of piggery waste

Summary A laboratory study was conducted to determine effects of the continuous mesophilic anaerobic digestion of raw pig manure in two types of enteropathogenic microorganisms, bacterial and viral. Faecal coliforms (indigenous to pig manure) and coliphage f2 (ATCC 15766 B1) were used as a model for some indigenous enteropathogenic microorganisms. The study was completed with laboratory survival experiments in lagoon stabilization of raw pig manure, for both models. Experiments for f2 survival in cell-free synthetic medium were also carried out. The results show that the anaerobic digestion process is more effective in eliminating viral than bacterial particles. Some parameters related to the ultimate biogas yield and kinetics were also determined. Lagoon stabilization of raw pig manure provides a more suitable environment for the removal of faecal coliforms than does anaerobic digestion. Finally, it was concluded that volatile fatty acids appeared to be responsible for the elimination of faecal coliforms. The agent that causes f2 inactivation is not well identified, although in some cases it could be NH₃ that seems to act as a vircidal agent.Correspondence to: J. Mata-Alvarez     

The effects of detergents on anaerobic digestion

Summary The anionic detergent sodium dodecylbenzene sulphonate (SDBS) inhibited mesophilic fermentation in anaerobic digesters. Total gas production and methanogenesis from glucose were reduced to half maximal rates at between 20 and 50 ppm SDBS during the initial phase of digestion, and over 20 days the pH declined from 7.4 to 6.0 in inhibited cultures. As well as accentuating the accumulation of propionic, iso-butyric and iso-valeric acids, a most remarkable effect of this anionic detergent observed only at high concentrations (100 ppm) was to divert the pathway of fermentation with transient accumulation of ethanol. Methanogenesis from cellulose was also inhibited. In thermophilic populations degrading glucose, SDBS was less toxic, and ethanol was not produced. Both the non-ionic detergent Tergitol (nonyl phenyl polyethylene glycol ether) and soap were virtually without effect on the mesophilic anaerobic digestion of glucose.     

Isolation of an anaerobic cellulolytic mixed culture

Summary Isolation and enrichment cultures were made for anaerobic cellulose utilizing micro-organisms from non-ruminant sources. Stable mixed cultures were developed which degraded pure cellulose (wet-milled filter paper) in a defined mineral salts medium. Components of the mixed cultures lost viability in monoculture when grown on cellulose. Growth on cellulose was stimulated at low oxygen concentrations, when increased cellulase activity and increased volatile fatty acid production occurred.Low concentrations (0.1–3 mM) of cellobiose, and to a lesser extent, glucose stimulated solubilization of cellulose by the cultures, but higher concentrations had an inhibitory effect.Growth on cellulose was accompanied by production of acetic, propionic and butyric acids. The production and profile of the acids was stable and characteristic of the culture. When an open nonaseptic fermentation was employed, the fatty acid profile was variable and also included valeric acid.     

The utilization of short-chain monocarboxylic acids as carbon sources for the production of gamma-linolenic acid by Mucor strains in fed-batch culture

The Fischer-Tropsch reaction water, which contains C₂ to C₅ monocarboxylic acids, generated as a co-product of the Sasol industrial oil-from-coal process, constitutes a potential cheap carbon substrate for the production of gamma-linolenic acid (GLA) by selced Mucor species. Three strains of Mucor were each grown in an air-lift reactor operated in a fed-batch, pH-stat mode under N-limitation with a mixture of C₂ to C₅ monocarboxylic acids as both pH titrant and carbon source. The production of GLA from this substrate was evaluated. Growth typically resulted in the rapid assimilation of acetic, n-butyric and n-valeric acids. Although propionic, iso-butyric and iso-valeric acids were assimilated to varying degrees, these acids accumulated in the culture. Mucor circinelloides CBS 203.28 gave the best results in that it assimilated 36% to 100% of each acid, had a biomass yield coefficient of 0.3 (calculated on acids utilized), and contained 28% crude oil, 84% of which comprised neutral lipids with a GLA content of 14.4%, giving 33 mg GLA/g biomass. GLA accumulation coincided with a decrease in the stearic-acid content of the neutral-lipid fraction. The results were comparable with previous results obtained with acetic acid and glucose as sole carbon sources, demonstrating the feasibility of producing GLA from the above mixture of organic acids.     

The effect of high and low dietary crude protein and inulin supplementation on nutrient digestibility, nitrogen excretion, intestinal microflora and manure ammonia emissions from finisher pigs

A 2 × 2 factorial experiment was performed to investigate the interaction between a high- and low-crude-protein (CP) diet (200 v. 140 g/kg) and inulin supplementation (0 v. 12.5 g/kg) on nutrient digestibility, nitrogen (N) excretion, intestinal microflora, volatile fatty acid (VFA) concentration and manure ammonia emissions from 24 boars (n = 6, 74.0 kg live weight). The diets were formulated to contain similar concentrations of digestible energy and lysine. Pigs offered the high-CP diets had a higher excretion of urinary N (P < 0.001), faecal N (P < 0.01) and total N (P < 0.001) than the pigs offered the low-CP diets. Inulin supplementation increased faecal N excretion (P < 0.05) and decreased the urine N : faeces N ratio (P < 0.05) compared with the inulin-free diets. There was no effect (P > 0.05) of dietary treatment on N retention. There was an interaction (P < 0.05) between dietary CP concentration and inulin supplementation on caecal Enterobacteria spp. Pigs offered the diet containing 200 g/kg of CP plus inulin decreased the population of Enterobacteria spp. compared to those with the inulin-supplemented 140 g/kg CP diet. However, CP level had no significant effect on the population of Enterobacteria spp. in the unsupplemented diets. Inulin supplementation increased caecal Bifidobacteria (P < 0.01) compared with the inulin-free diets. There was no effect of inulin supplementation on VFA concentration or intestinal pH (P > 0.05). Pigs offered the 200 g/kg CP diets had higher (P < 0.05) manure ammonia emissions from 0 to 240 h of storage than pigs offered the 140 g/kg CP. In conclusion, inulin supplementation resulted in an increase in Bifidobacteria concentration and a reduction in Enterobacteria spp. at the high CP level indicating that inulin has the ability to beneficially manipulate gut microflora in a proteolytic environment.