Genome-wide immunity studies in the rabbit: transcriptome variations in peripheral blood mononuclear cells after in vitro stimulation by LPS or PMA-Ionomycin.

BACKGROUND: Our purpose was to obtain genome-wide expression data for the rabbit species on the responses of peripheral blood mononuclear cells (PBMCs) after in vitro stimulation by lipopolysaccharide (LPS) or phorbol myristate acetate (PMA) and ionomycin. This transcriptome profiling was carried out using microarrays enriched with immunity-related genes, and annotated with the most recent data available for the rabbit genome. RESULTS: The LPS affected 15 to 20 times fewer genes than PMA-Ionomycin after both 4 hours (T4) and 24 hours (T24), of in vitro stimulation, in comparison with mock-stimulated PBMCs. LPS induced an inflammatory response as shown by a significant up-regulation of IL12A and CXCL11 at T4, followed by an increased transcription of IL6, IL1B, IL1A, IL36, IL37, TNF, and CCL4 at T24. Surprisingly, we could not find an up-regulation of IL8 either at T4 or at T24, and detected a down-regulation of DEFB1 and BPI at T24. A concerted up-regulation of SAA1, S100A12 and F3 was found upon stimulation by LPS. PMA-Ionomycin induced a very early expression of Th1, Th2, Treg, and Th17 responses by PBMCs at T4. The Th1 response increased at T24 as shown by the increase of the transcription of IFNG and by contrast to other cytokines which significantly decreased from T4 to T24 (IL2, IL4, IL10, IL13, IL17A, CD69) by comparison to mock-stimulation. The granulocyte-macrophage colony-stimulating factor (CSF2) was by far the most over-expressed gene at both T4 and T24 by comparison to mock-stimulated cells, confirming a major impact of PMA-Ionomycin on cell growth and proliferation. A significant down-regulation of IL16 was observed at T4 and T24, in agreement with a role of IL16 in PBMC apoptosis. CONCLUSIONS: We report new data on the responses of PBMCs to LPS and PMA-Ionomycin in the rabbit species, thus enlarging the set of mammalian species for which such reports exist. The availability of the rabbit genome assembly together with high throughput genomic tools should pave the way for more intense genomic studies for this species, which is known to be a very relevant biomedical model in immunology and physiology.

Quantitative feed restriction rather than caloric restriction modulates the immune response of growing rabbits.

BACKGROUND: Short-term feed restriction strategies are used in rabbits to reduce postweaning digestive disorders, but little is known about the involvement of the immune system in these beneficial effects. OBJECTIVE: In the present study, the consequences of feed and energy restriction on immune response were investigated. METHODS: At weaning, 320 male and female rabbits were assigned to 4 groups differing in dietary digestible energy (DE) concentrations and intake levels: a low-energy ad libitum-feed (LE100) group, a low-energy restricted-feed (LE75) group, a high-energy ad libitum-feed (HE100) group, and a high-energy restricted-feed (HE75) group. The high-energy groups consumed 10.13 MJ DE/kg of feed, whereas the low-energy groups consumed 9.08 MJ DE/kg (formulated values). Intake amounts for the restricted groups were 75% those of the ad libitum groups. Rabbits consumed these diets until age 63 d, after which they consumed feed ad libitum for 9 d. Ten rabbits per group and per age were killed at ages 42, 50, 63, and 72 d. Spleens and appendixes were weighed; Peyer's patch surface area was determined by image analysis; plasma total immunoglobulin (Ig) G and anti-ovalbumin IgG; and fecal and plasma IgA concentrations were determined by ELISA; and ileal expressions of cytokines were measured by quantitative reverse transcriptase-polymerase chain reaction at ages 50 and 63 d. RESULTS: The relative weight and size of the lymphoid organs were not affected by treatments. Concentrations of plasma total IgA (-41% at 63 d and -29% at 72 d), IgG (-22% at 72 d), and anti-ovalbumin IgG (-41% at 63 d) were lower with feed restriction. Fecal IgA concentrations were lower with quantitative restriction (-40%, -52%, and -65% at age 42, 50, and 63 d, respectively) and energy restriction (-56%, -46%, and -73% at ages 50, 63, and 72 d, respectively). Feed-restricted rabbits tended to have greater expressions of interleukin (IL) 1ß and IL-2 and lower expressions of tumor necrosis factor α (P < 0.1). CONCLUSION: These results demonstrated that, in rabbits, restriction and, to a lesser extent, dietary energy concentration modulate gut immunity.

A multi-omics dataset of the response to early plant polysaccharide ingestion in rabbits.

The transition from a milk-based diet to exclusive solid feeding deeply modifies microbiota-host crosstalk. Specifically, early ingestion of plant polysaccharides would be one of the main nutritional components to drive host-microbiota-interaction. To capture the effects of polysaccharides early-life nutrition (starch vs rapidly fermentable fiber) on the holobiont development, we investigated on the one hand the gut bacteriome and metabolome and on the other hand the transcriptome of two host gut tissues. Rabbit model was used to study post-natal co-development of the gut microbiota and its host around weaning transition. The assessment of the microbial composition of the gut appendix together with the caecum was provided for the first time. Gene expression signatures were analyzed along the gut (ileum and caecum) through high-throughput qPCR. The data collected were completed by the analysis of animal growth changes and time-series assessment of blood biomarkers. Those accessible and reusable data could help highlight the gut development dynamics as well as biological adaptation processes at the onset of solid feeding.

Early Introduction of Plant Polysaccharides Drives the Establishment of Rabbit Gut Bacterial Ecosystems and the Acquisition of Microbial Functions.

In mammals, the introduction of solid food is pivotal for the establishment of the gut microbiota. However, the effects of the first food consumed on long-term microbiota trajectory and host response are still largely unknown. This study aimed to investigate the influences of (i) the timing of first solid food ingestion and (ii) the consumption of plant polysaccharides on bacterial community dynamics and host physiology using a rabbit model. To modulate the first exposure to solid nutrients, solid food was provided to suckling rabbits from two different time points (3 or 15 days of age). In parallel, food type was modulated with the provision of diets differing in carbohydrate content throughout life: the food either was formulated with a high proportion of rapidly fermentable fibers (RFF) or was starch-enriched. We found that access to solid food as of 3 days of age accelerated the gut microbiota maturation. Our data revealed differential effects according to the digestive segment: precocious solid food ingestion influenced to a greater extent the development of bacterial communities of the appendix vermiformis, whereas life course polysaccharides ingestion had marked effects on the cecal microbiota. Greater ingestion of RFF was assumed to promote pectin degradation as revealed by metabolomics analysis. However, transcriptomic and phenotypic host responses remained moderately affected by experimental treatments, suggesting little outcomes of the observed microbiome modulations on healthy subjects. In conclusion, our work highlighted the timing of solid food introduction and plant polysaccharides ingestion as two different tools to modulate microbiota implantation and functionality. IMPORTANCE Our study was designed to gain a better understanding of how different feeding patterns affect the dynamics of gut microbiomes and microbe-host interactions. This research showed that the timing of solid food introduction is a key component of the gut microbiota shaping in early developmental stages, though with lower impact on settled gut microbiota profiles in older individuals. This study also provided in-depth analysis of dietary polysaccharide effects on intestinal microbiota. The type of plant polysaccharides reaching the gut through the lifetime was described as an important modulator of the cecal microbiome and its activity. These findings will contribute to better define the interventions that can be employed for modulating the ecological succession of young mammal gut microbiota.

Impact of feed restriction and fragmented feed distribution on performance, intake behaviour and digestion of the growing rabbit.

Postweaning feed restriction preserves rabbit digestive health after weaning, but the underlying physiological mechanisms are not yet understood. To elucidate whether the feeding intake pattern modification related to feed restriction might be involved, we studied the effects of both feed intake quantity and intake frequency. Animals were allotted at weaning (28 d old) in a 2 × 2 factorial design: feed intake quantity (AL = ad libitum vs R = 75% of AL) and fragmented feed distribution (FFD) (1 vs 13 distributions), thus forming four groups (AL1, AL13, R1 and R13). New Zealand White growing rabbits were used from weaning to slaughter (70 d old), to analyse mortality, morbidity, performance, intake behaviour, digestion and microbial activity. Seven days after starting feed restriction (35 d old, group R1), rabbits consumed 44% of the feed within 2 h, 65% in 4 h and in 7 h over 95%. Over the 28-70 d period, mortality was low (5.3%) while morbidity averaged 18.5% and neither was affected by treatment. However, FFD tended to decrease the morbidity rate during the first 14 days after weaning (P = 0.06). Feed conversion (28-70 d) was improved by restriction (+15%, P < 0.001) and by FFD (+5%, P < 0.001). Nutrient digestibility was improved by restriction (+10% for energy, P < 0.01), but not by FFD. Fragmented feed distribution led to a lower stomachal pH, in the antrum (1.48 vs 2.13, P < 0.001) and in the fundus (1.52 vs 2.63, P < 0.001), while a higher pH was found in the caecum (6.07 vs 5.86, P < 0.001). Butyrate proportion in the caecum was reduced by four units for restricted groups. Fragmented feed distribution reduced the caecal VFA concentration by 23% within restricted rabbit groups only. A similar interaction between intake level and FFD was observed for fibrolytic activity (cellulase and xylanase). The diversity of caecal bacterial community was not modified by either of the two factors studied. Globally, fragmented meals have no major impacts on the caecal microbial activity, diversity, and thus would not be implicated in the better resistance of restricted rabbit to digestive troubles.

Comparison of the archaeal community in the fermentative compartment and faeces of the cow and the rabbit.

The archaeal community in the fermentative compartment and faeces of the cow and the rabbit were compared by analysis capillary electrophoresis single-stranded conformation polymorphism (CE-SSCP) profiles of 16S rRNA genes. Ruminal and faecal contents were sampled in five cows for three weeks. Hard and soft faeces were collected in 14 rabbits for three consecutive weeks and caecal contents were sampled in the third week. The archaeal community differed according to the host species (ANOSIM-R=0.53 and 0.72 respectively for the comparison of the fermentative compartments and faeces; P<0.001) and to the location within the digestive tract of both species (ANOSIM-R=0.37, 0.52 respectively for the cow and the rabbit; P<0.001). In both species, the archaeal community of the digestive tract was stable over weeks and varied very little between individual animals. The structure (NS) and the richness index (9.9+/-2.7, 10.1+/-3.1 respectively, NS) of the archaeal community were similar for the caecal content and the soft faeces which permitted to use the latter as a representative indicator.

Molecular analysis of the bacterial community in digestive tract of rabbit.

This work aimed to study the stability over time of the bacterial community in caecum and faeces of the rabbit (diversity index and structure) without experimental disturbance and to evaluate its relationships with environmental parameters. Soft and hard faeces of 14 rabbits were sampled for 5 weeks while caecal content was sampled on the 3rd week (by surgery) and the 5th week (at slaughter). Bacterial communities were assessed by studying CE-SSCP profiles of 16S rRNA genes fragments. Redox potential, pH, NH3-N concentration and volatile fatty acid concentrations were measured in the caecum. Data showed that bacterial communities of soft and hard faeces barely differed from that of the caecum (ANOSIM-R<0.25; p<0.05). Without disturbance, the bacterial communities of faeces were stable over time (ANOSIM-R<0.25; p<0.001). However, the bacterial communities of caecum and faeces were affected by the surgery (ANOSIM-R=0.22-0.33; p<0.001). The caecal content was an acidic (pH=6.03+/-0.33) and an anaerobic environment (redox potential=-160+/-43 mV). Only the redox potential was correlated with the diversity index of the bacterial community of the caecum (R(2)=0.35; p<0.05) and no environmental parameters were correlated to its structure.

Data set on early feed intake and growth performances of rabbits fed during the suckling period with pellets differing in diameter or compression rate using a double-choice testing design.

Weaning is a critical period for the health of rabbits, with a high sensitivity to digestive diseases. Allowing early consumption of solid feed in the nest of the suckling rabbit could help to maintain its health around weaning. In general, previous studies have focused on feed intake of rabbits when they are able to leave the nest, i.e. around 16 days. Herein, we provide a unique dataset of the dynamics of the onset of feed intake in suckling rabbits from 8 days to weaning. We quantified the solid feed intake behaviour and determined the dietary preferences for pellets according to their physical properties using nine pellets differing in diameter or compression rate. Additionally to the data provided in Paës et al. [1] we provide (i) the description of the nine pellets processing (ii) the description of the 3 point-scale system for nest quality evaluation, (iii) details on the device used to provide pellets in the nest, (iv) milk intake data and milk intake curve calculation and (v) pellet intake data according to physical characteristics.

Early Introduction of Solid Feeds: Ingestion Level Matters More Than Prebiotic Supplementation for Shaping Gut Microbiota.

Early introduction of a nutritional substrate is a promising biomimetic strategy for controlling the implantation of the microbiota and preserving the health of young animals. In this study, we provided experimental solid substrate in a gel form to stimulate suckling rabbits' intake and to investigate its effects on microbiota implantation and colonization. All the rabbits had access to solid feed outside the nest as of 15 days of age. Except for the control group, rabbits were offered starter feed gels inside the nests from 3 to 18 days of age. These gels were either free of additives (AF_GEL) or contained 4% of fructo-oligosaccharides (FOS_GEL) or 4% of mannan-oligosaccharides and ß-glucans mixtures (MOS_GEL). The cecal content of 160 rabbits was sampled at 18, 29, 38, and 57 days of age and analyzed using 16S rRNA gene sequencing. Pups consumed an average of 3.95 ± 1.07 g of starter feed gel with a higher intake when it was supplemented with fructo-oligosaccharides (+1.2 g; P < 0.05). Starter feed gel consumption increased the ensuing intake of pellets (+17 g from 15 to 21 days; P < 0.05). Alpha-diversity indexes were similar between groups and prebiotic supplementation did not induce a clear shift in microbiota pattern. Conversely, when considering rabbits that consumed more starter feed, the highest proportions of bacteria with plant-degrading abilities, such as species from the Lachnospiraceae and Ruminococcaceae families, were observed at 18 days of age. However, fermentative activities were not affected by starter feed intake at 29, 38, and 57 days of age. By providing comprehensive results on the regulation of microbial community structure at the onset of solid feed intake, this research paves the way for further studies on digestive ecosystem maturation.

Diversity and Co-occurrence Pattern Analysis of Cecal Microbiota Establishment at the Onset of Solid Feeding in Young Rabbits.

This study aimed to evaluate how the feeding strategy of rabbit kits at the onset of solid feed intake could affect ecological diversity and co-occurrence patterns of the cecal bacterial community. From birth to 18 days of age kits were exclusively milk-fed, and between 18 and 35 days the young rabbits also had access to solid feed. After weaning at (35 days), young rabbits were exclusively fed solid feed. Three experimental feeds were used: a high concentrate diet [H: 10.16 MJ digestible energy (DE)/kg and 15.3% crude protein (CP)], a low concentrate diet (L: 9.33 MJ DE/kg and 14.7% CP) and a reproductive female diet (R: 10.57 MJ DE/kg and 17.3% CP). The rabbit kits (n = 357) were divided into three groups, differing by the diet received during two periods: from 18 to 28 and from 28 to 49 days of age. In the groups LL and HH, rabbit kits were fed L or H diets, respectively, during both periods. Kits in the group RL received feeds R and L from 18 to 28 and 28 to 49 days of age, respectively. Cecal bacterial communities of 10 rabbits per group were carried out at 18, 28, 35, 43 and 49 days of age by MiSeq Illumina sequencing 16S rRNA encoding genes. Between 18 and 28 days of age, solid feed intake was higher in the group RL compared to the other two groups (+24%; P < 0.01). Overall, 13.4% of the OTUs detected were present in the cecal ecosystem from 18 to 49 days old, whereas 17.4% were acquired with the onset of solid feeding and kept from 28 days on. Exclusive milk consumption constrains the bacterial community toward a similar structure but high phylogenetic beta-diversity. Introduction of solid feed induced a sharp change of microbial community structure and decreased phylogenetic diversity. A strong relationship in bacterial community network occurred only from 43 days on. Our feeding strategy at the onset of solid feed ingestion exhibited only a moderate effect on the microbial community structure (P = 0.072), although the LL group seemed to reach faster maturity compared to the two other groups.

Potential core species and satellite species in the bacterial community within the rabbit caecum.

A bacteria library was constructed from the caecum of a rabbit maintained under standard conditions. The complete gene 16S rRNA gene was sequenced. The 228 clones obtained were distributed in 70 operational taxonomic units (OTUs). The large majority of the OTUs were composed of one or two clones and seven OTUs contained half of the sequences. Fourteen sequences had high similarity to the sequence already registered in databases (threshold of 97%). Only one of these sequences has been identified as Variovorax sp. (99% identity). Units were distributed mainly (94%) in the Firmicutes phylum. Three sequences were related to Bacteroidetes. Nine clusters were defined in the phylogenic tree. A great diversity of caecal bacteria of the rabbit was shown. Half of the sequences generated in this library were distributed in the phylogenetic tree near the sequences characterized previously in rabbit caecum (potential core species), and the other half of the sequences were well separated (satellite species).

Testing the efficacy of medium chain fatty acids against rabbit colibacillosis.

Enteropathogenic Escherichia coli (EPEC) represents a major cause of lethal diarrhea in young mammals. Although the pathogenicity mechanisms of EPEC are now well understood, the intrinsic and environmental factors that control the expression of EPEC virulence remain largely unknown. In the rabbit, suckling reduces pups' sensitivity to EPEC infection. Hence, we have hypothesized that uncharacterized factors present in doe milk may mediate this protection. Medium chain fatty acids (MCFA), known to possess antimicrobial properties, are highly abundant in doe milk. We demonstrate that caprylic acid exhibits a clear bacteriostatic effect in vitro against the rabbit EPEC strain E22 (O103:H2:K-), in a dose-dependent manner. In vivo, the dietary inclusion of triglycerides of MCFA did not however reduce the sensitivity of young rabbits challenged with this EPEC strain. The mortality and fecal excretion of EPEC were not reduced, and the bacterial adhesion to ileum was not inhibited. Amount of MCFA reaching the ileal level might have been too low and/or their association to other milk antimicrobials may have been required to observe a positive effect on disease evolution in a context of a highly virulent challenge.

Postnatal development of the rabbit caecal microbiota composition and activity.

This study describes the development of the rabbit caecum microbiota and its metabolic activities from the neonatal (day 2) until the subadult period (day 70). The caecal microbiota was analysed using 16S rRNA gene approaches coupled with capillary electrophoresis single-stranded conformation polymorphism (CE-SSCP) and qPCR. At day 2, rabbits harboured population levels up to 8.4, 7.2 and 7.4 log(10) copy number g(-1) full caecum of the total bacteria, Bacteroides-Prevotella and Firmicutes groups, respectively. These populations reached their maximum levels from day 14 for Firmicutes groups (10.8 log(10) copy number g(-1) caecal content) and day 21 (11.4 and 10.7 log(10) copy number g(-1) caecal content of the total bacteria and the Bacteroides-Prevotella group, respectively). The archaeal population could be detected only from day 7 onwards (5.5 log(10) copy number g(-1) full caecum) and reached its maximum level at day 35 (7.4 log(10) copy number g(-1) caecal content). Similarity analysis, diversity calculation and quantitative evaluation of the stability of bacterial community CE-SSCP profiles provided some evidence that the caecal microbiota develops progressively from a simple and unstable community after birth into a complex and climax community in subadult rabbits. Meanwhile, the microbial activity evolved with the progressive decrease of the propionate/butyrate ratio towards a rabbit-specific value <1.

Maternal milk contains antimicrobial factors that protect young rabbits from enteropathogenic Escherichia coli infection.

Enteropathogenic Escherichia coli (EPEC) colibacillosis represents a major cause of lethal diarrhea in young children in developing countries. EPEC strains also infect numerous mammal species and represent a major economical problem in rabbit industry. Protection against this pathogen is a challenging goal both in humans and in other mammal species. Despite a good knowledge of the pathogenicity mechanisms of EPEC, the intrinsic and environmental factors that control the expression of EPEC virulence in mammals remain unknown. For instance, the exacerbated sensitivity of young mammals to EPEC infection is still unexplained. Our goal was to investigate if age or other factors, like milk consumption, could be determinants that trigger the disease. We used rabbits as an animal model to study the role of milk in the sensitivity to an EPEC infection. Weaned and suckling rabbits were orally inoculated with EPEC strain E22 (O103:H2:K-) at 28 days of age, and the evolution of the disease was investigated in the two groups. In addition, in order to better characterize the interactions between milk and EPEC, we determined in vitro bacterial growth and the abilities of EPEC cells to adhere to epithelial cells in the presence of milk. Our results demonstrate a protective role of milk in vivo in association with in vitro antibacterial activity. These effects are independent of the presence of specific anti-EPEC antibodies.

An early stimulation of solid feed intake slightly influences the morphological gut maturation in the rabbit.

The impact of dietary factors on the gut morphological maturation is poorly documented in rabbits. The weights of the digestive segments as well as the morphology of villi and crypts along the small intestine were analysed weekly from day 14 till day 49, in two rabbit groups weaned at either 21 (W21 group, n = 12 litters) or 35 days (W35 group, n = 12 litters) of age. From 21 till 35 days, the W21 group ate 57% more solid feed than the W35 group (P < 0.01), and presented slighter body weights from day 28 till day 49 (-9%, P < 0.05). Tissue weights of the empty digestive segments, as expressed relative to the body weights, were higher in the W21 than in the W35 group from day 28 till day 49 (P < 0.001), whereas absolute tissue weights appeared similar (except for the proximal colon). From day 28 to day 49, small intestinal villi grew in height and surface area (P < 0.05) whereas the crypts deepened. Villous height followed a proximo-distal decreasing gradient from the duodenum to the ileum (P < 0.05) from day 28 onward. The villous height to width ratio changed with the beginning of significant solid feed intake: from a thin shape until day 21, villi became wider from day 28 on. The effect of weaning age on mucosal morphology was insignificant, except for the jejunal crypts whose surface area and depth were higher in the W21 group. The present results showed that morphological changes in the digestive tract of young rabbits were weakly influenced by an early stimulation of solid feed intake.

Nutritional and sanitary statuses alter postweaning development of caecal microbial activity in the rabbit.

The postweaning development of caecal microbial activity was studied in the rabbit according to the sanitary status (conventional "C" vs. specified pathogen-free "SPF") and the nutritional status (standard-fibre "SF" vs. deficient-fibre "DF" diet). The two diets were distributed ad libitum from weaning (28 days) to 70 days of age, respectively, to 80 C and 72 SPF rabbits. From 28 to 42 days, the volatile fatty acids concentration in the caecum (tVFA) of C rabbits was 50 mM/L and increased by 46% between 42 and 56 days, without interactions with the diet effect. In parallel, the bacterial fibrolytic activity decreased for xylanase and CMCase (-32% and -60%, respectively, P<0.05), while pectinase activity decreased more regularly from 28 to 70 days (-28%, P<0.05). At weaning, tVFA was similar among C or SPF rabbits, while at 70 days, it decreased by 23% for SPF and increased in C group (+31%). Cellulasic and hemicellulasic activity of bacteria were two to three times lower, respectively, in SPF rabbits compared to conventional ones. No interaction was detected between sanitary and nutritional status at 70 days of age for the caecal fermentative activity. With the FD diet, tVFA decreased by 10%, while butyrate proportion increased by 37% (at 70 days), whatever the sanitary status. In 70-day-old rabbits (C or SPF group), pectinasic activity was reduced by 30% when rabbits were fed the FD compared to the SF one.

Inter-relationship of microbial activity, digestion and gut health in the rabbit: effect of substituting fibre by starch in diets having a high proportion of rapidly fermentable polysaccharides.

Caecal microbial activity, digestion and gut health were analysed in the young rabbit, in response to fibre substitution by starch, in diets with high proportions of rapidly fermentable polysaccharides (pectins+hemicelluloses:acid-detergent fibre (ADF) ratio of 1.7). A range of five diets corresponding to a 60 % linear reduction of the ADF level (230 to 92 g ADF/kg) without changes in the fibre quality, and to a corresponding linear increase in dietary starch, was given ad libitum to young rabbits from 18 d until 70 d of age. A one half reduction of the ADF level resulted in a sharp increase in energy digestibility (+25 units), associated with a lower feed intake (-35 %) and to an increase of the mean retention time (+6 h) in the whole digestive tract. Despite large variations in the fibre intake (20 to 59 g ADF/d), the fibre digestive efficiency remained similar among the five diets. Starch ileal concentrations were low after 4 weeks of age (<5 %), and variations with age were significant when the dietary starch level was over 19 %. A 65 % lower biomass production was measured when the ADF level progressed from 230 to 165 g/kg, and no precise relationship was found with fermentative activity. Reducing the fibre intake led to a linear decrease of caecal volatile fatty acids concentrations, and to higher pH and NH3 levels. An increased occurrence of mortality by diarrhoea was registered with the lowest fibre intake. It can be concluded that a sufficient supply of fibre, with high proportions in rapidly fermentable polysaccharides, stimulates the maturation of microbial activity and reduces the occurrence of diarrhoea.

Digestive tract development in rabbit according to the dietary energetic source: correlation between whole tract digestion, pancreatic and intestinal enzymatic activities.

The developmental changes of both pancreatic and intestinal enzymes and the influence of dietary composition on enzyme activities were followed in suckling and weaning rabbits. In addition, whole tract digestibility of nutrients was recorded in response to two dietary energetic sources. Rabbits were fed ad libitum either a low fat and high starch diet (group LF), or a high fat and high fibre diet (group HF) between d 32 and d 42, with both groups receiving a growing finishing diet thereafter. Before weaning (d 32) nutrient digestion was high (>75% for organic matter, protein or fat), and then decreased sharply, except for fat. Between d 32 and d 42, digestion in the HF group was 7.5 and 4.6% lower, respectively, for organic matter and protein, while fibre and fat digestion was higher (+14.0 and +5.0%, respectively). Between d 25 and d 42 of age, pancreatic-specific activities of trypsin and chymotrypsin did not change while those of amylase and lipase increased by 1.5- and 76- fold (P<0.05), respectively. However, total activities and relative activities expressed on a LW basis were increased after weaning as a main consequence of a specific increased organ weight and pancreatic protein content. Relative activities of trypsin and chymotrypsin increased by 63 and 56% (P<0.01) after weaning, respectively. Total activities of pancreatic enzymes measured in the total small intestinal contents increased during the same period, but the range of variations was lower than those measured in the pancreatic gland. Total activities of lipase, trypsin and chymotrypsin measured in the small intestine contents were significantly correlated with pancreas enzyme potentialities. Total small intestine activity of lipase was 58% higher (P<0.001) in HF than in LF group while the other pancreatic and intestinal enzyme activities measured were not influenced by the energetic sources of the diet. Decreased digestibility of organic matter and protein observed with the HF diet could not be related to changes in pancreatic or intestinal enzymatic profiles and may be more dependent on quality of dietary ingredients.