Introduction. Consumption of citrus fruits and their juice has strongly grown in the last few decades. However, at the present time, the traceability of these fruits is only documentary. In case of doubt or fraud, no standardized analysis makes it possible to discriminate or determine the geographical origin of culture of the fruit. Materials and methods. A method of bacterial ecology, PCR-DGGE, was used to characterize the bacterial flora of clementines imported into France from Spain and Morocco in order to show that there is a relation between the bacterial communities of the fruits and their geographical origins. The principle rests on the determination of specific biological markers for a given region. Protocols of microbial extraction and DNA amplification were optimized. Results. DGGE profiles analyzed by multivariate analysis permitted us to distinguish microbial communities from different origins. Conclusion. We propose the PCR-DGGE method as a new tool for traceability that provides citrus products with a unique bar code and makes it possible to trace back the citrus fruit to their original location.

Six cannulated wether sheep weighing 57 (s.d. 4·3) kg were used to investigate the susceptibility of unprotected and protected n-3 polyunsaturated fatty acids from different sources to biohydrogenation in the rumen, their uptake into plasma and effects on ruminal metabolism. The sheep were assigned to one of six dietary treatments formulated to have a similar fatty acid content (60 g/kg DM) and containing: linseed oil (LO), linseed oil absorbed into vermiculite (VLO), formic acid-formaldehyde treated whole linseed (FLS), fish oil (FO), fat encapsulated fish oil (PFO) or a mixture of fish oil and marine algae (1: 1 on an oil basis; AF), in six periods of 28 days duration in a Latin-square design. Biohydrogenation of C20:5 (n-3) and C22:6 (n-3) was high in FO at approximately 870 g/kg, but reduced to 625 and 625 g/kg respectively for PFO, and 769 and 601 g/kg respectively for AF. Ruminal biohydrogenation of C18:3 (n-3) was similar across treatments based on linseed, averaging 860 g/kg, but C18:2 (n-6) was lower (P < 0·05) in animals given VLO or FLS at 792 and 837 g/kg respectively, compared with LO (907 g/kg). Duodenal flow of C18:1 trans in animals given any of the diets containing fish oil averaged 8·4 g/day compared with 2·8 g/day in animals given diets based on linseed (P < 0·001), whilst cis-9, trans-11 conjugated linoleic acid was not significantly different among treatments. Plasma C20:5 (n-3) and C22:6 (n-3) proportions were highest in animals given the AF diet (11·8 and 8·2 g per 100 g of the total fatty acids respectively) and lowest in animals given LO (2·8 and 2·7 g per 100 g of the total fatty acids respectively; P < 0·001). By contrast, plasma C18:3 (n-3) proportions were highest in animals given the LO or VLO diets at approximately 6·9 g per 100 g of the total fatty acids, and lowest in the AF treatment at 0·9 g per 100 g (P < 0·001). Duodenal non-ammonia-N flow was similar among treatments at 21·0 g/day except in animals given FLS which had the highest flow (25·9 g N per day; P < 0·01). Microbial N flow was also similar among treatments whilst microbial efficiency (g N per kg OM truly degraded in the rumen) was higher (P < 0·05) in animals given FLS than LO, FO or AF. By contrast, ruminal fibre digestion was higher (P < 0·05) in animals given LO or FO than those offered VLO, FLS, PFO or AF. In conclusion, compared with linseed oil, absorption of linseed oil into vermiculite improved duodenal flow but not plasma levels of C18:3 (n-3), whilst formic acid-formaldehyde treatment of linseed had little effect on protecting C18:3 (n-3) in the rumen, although duodenal non-ammonia nitrogen flow and microbial efficiency were improved. Compared with fish oil, the provision of marine algae or fat encapsulated fish oil resulted in a lower biohydrogenation of C22:6 (n-3) and C20:5 (n-3), and an increased duodenal flow and plasma concentration and offers the potential to favourably manipulate the n-3 fatty acid composition of sheep meat.

The origin of duodenal purine bases (PB) was studied in a digestion experiment with four heifers, cannulated in the rumen and duodenum, which received a basal concentrate (152 g crude protein (CP) per kg dry matter (DM)) together with barley straw (85: 15 fresh weight basis) or the same concentrate supplemented with soya-bean meal, carbohydrate-treated soya-bean meal, maize gluten meal or fish meal to increase its protein content to 192 g/kg DM. Tr eatments were assigned to the four animals in five experimental periods according to an incomplete Latin-square design. Each 30-day period included 20 days of change-over adaptation and 10 days of experimental measurements. The flow of digesta entering the duodenum was estimated using Yb and acid-detergent insoluble ash as indigestible markers according to a double-marker system and microbial nitrogen (N) and PB were labelled with 15N infused into the rumen. The proportion of duodenal PB of microbial origin estimated from 15N enrichment of PB-N averaged 0·66 (s.e. 0·029) and did not differ between treatments nor when protozoa or bacteria associated with liquid (LAB) and solid (SAB) fractions were used as a reference sample. On average microbial contribution to duodenal non-ammonia N was higher when estimated from the PB/N ratio than from 15N (0·67 v. 0·55 (s.e. 0·015)) although differences were small and not significant when LAB was the reference sample (0·58 v. 0·52 (s.e. 0·018)) reflecting the higher PB/N ratio of this fraction compared with SAB and protozoa (2·04 v. 1·65 and 1·60 (s.e. 0·04) mmol/g). Considering only the duodenal PB of microbial origin resulted in estimates of microbial N synthesis from the PB/N ratio of SAB similar to those derived from 15N enrichment of both bacterial fractions (12·9 v. 13·5 and 13·3 (s.e. 0·83) g/kg of organic matter apparently digested in the rumen OMADR)) but underestimated the values derived from LAB (9·9 g/kg OMADR). Regardless of the estimation method, neither the duodenal flow of microbial N nor the efficiency of microbial synthesis differed between treatments. These results suggest that a significant proportion of duodenal PB have a non-microbial origin which may lead to overestimation of microbial yield when PB are used as a marker. Differences in PB/N ratio between microbial fractions is another important factor to be considered.

The effect of carbohydrate supplementation on rumen utilization of ammonia-treated straw was studied in four rumen-cannulated ewes (42•0 (s.e. 2•80) kg live weight) in a Latin-square design. Diets were a 50: 50 mixture of ammoniated barley straw and a concentrate made up with different ratios of barley grain and citrus pulp, namely M1 (100: 0), M2 (66: 33), M3 (33: 66) and M4 (0: 100). Concentrates also included soya-bean meal and urea to make diets isonitrogenous. Diets were given in one meal in the morning at 0•8 kg/day. There were no significant differences among treatments in daily weighted means of rumen pH and concentration of volatile fatty acids. Acetate, propionate and butyrate molar proportions were not different among treatments, but proportions of isobutyrate (P < 0•05) and isovalerate (P < 0•10) were higher for M1 and M2. Ammonia concentration immediately before feeding was lower (P < 0•05) for M3 and M4 (72•1 and 51•3 mg/l) than for M1 and M2 (97•0 and 107•9 mg/ l). Total rumen bacterial concentration was higher (P < 0•05) with the highest barley proportion, whereas enzymatic activity against structural carbohydrates of particle-associated bacteria did not differ significantly among treatments. Digestibility of neutral-detergent fibre increased (P < 0•05) with the increase in the proportion of citrus pulp in the concentrate (0•647, 0•674, 0•684 and 0•693 for M1, M2, M3 and M4; s.e. = 0•0086). Urinary excretion of purine derivatives decreased linearly (P < 0•05) as the proportion of citrus pulp in the diet increased, while faecal excretion of purine bases was not significantly affected by dietary treatments.