A first group of three horses was given diet 1 (D1) allowing 1180 g per 100 kg body weight (BW) of a pelleted food rich in fibre (P1) and 556 g per 100 kg BW of straw during a 20-day period to allow for adaptation. A second group of four horses were given diet 2 (D2) allowing 1180 g per 100 kg BW of a pelleted food rich in cereals (P2) and 1000 g per 100 kg BW of meadow hay during the same period. Digesta was collected from the stomach, duodenum, jejunum, ileum, caecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, and small colon, and faeces were collected under general anaesthesia 2·5 h after the ingestion of the morning pelleted meal. The concentration of total anaerobic, cellulolytic and lactic acid-utilizing bacteria, lactobacilli and streptococci were determined in all these segments except for the duodenum, left ventral colon, right dorsal colon and small colon. D -/ Llactic acid, volatile fatty acids and pH were measured in all anatomic segments of the digestive tract (from stomach to small colon). The caecal concentration of total anaerobic bacteria was the lowest (7 ⋅ 9 5 10 7 colony-forming units (c. f. u. ) per ml), whereas that of the stomach was the highest (1·4 5 10 9 c. f. u. per ml) ( P < 0 ⋅ 001). Cellulolytic bacteria did not exceed 3·0 5 10 2 c. f. u. per ml in the ante-caecal segments whereas in the hindgut the average concentration was 5·3 × 10 5 c. f. u. per ml ( P < 0 ⋅ 001). Likewise, VFA concentrations were also greater in the large intestine (on average, 96·3 mmol/l v. 8·8 mmol/l in the ante-caecal segments) ( P < 0 ⋅ 001), confirming the limited extent of fibre degradation in these ante-caecal segments. Lactobacilli, streptococci and lactate-utilizing bacteria colonized all the digestive tract; the stomach and the small intestine tended to host the greatest numbers of these bacteria, which suggests a high interference of micro-organisms with the digestion of readily fermentable carbohydrates. Compared with the other ante-caecal segments, the stomach ecosystem seemed the most affected by the composition of the last pelleted meal ingested : the concentrations of lactobacilli and lactate-utilizing bacteria were higher ( P < 0 ⋅ 05) with P2. The lower concentration of D -/ L -lactate with P2 ( P < 0 ⋅ 05) was concomitant with a greater proportion of propionate ( P < 0 ⋅ 05), probably related to a greater fermentation of lactate. In the large intestine of horses given D2, cellulolytic bacteria tended to be lower, whereas VFA concentrations were higher ( P < 0 ⋅ 05). The lower [NDF/starch] ratio of D2 was probably less propitious for the proliferation of cellulolytic bacteria but was compensated by the higher cellulose intake brought by the hay.

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Background: Dirofilaria immitis, D. repens and Acanthocheilonema reconditum are the main causative agents of zoonotic canine filariosis. Methods: We developed a combined multiplex approach for filaria and Wolbachia detection using the 28S-based pan-filarial and 16S-based pan-Wolbachia qPCRs, respectively, involving a fast typing method of positive samples using triplex qPCR targeting A. reconditum, D. immitis and D. repens, and a duplex qPCR targeting Wolbachia of D. immitis and D. repens. The approach was complemented by a duplex qPCR for the differential diagnosis of heartworms (D. immitis and Angiostrongylus vasorum) and pan-filarial cox1 and pan-Wolbachia ftsZ PCRs to identify other filarial parasites and their Wolbachia, respectively. A total of 168 canine blood and sera samples were used to validate the approach. Spearmanʼs correlation was used to assess the association between filarial species and the strain of Wolbachia. Positive samples for both the heartworm antigen-test after heating sera and at least one DNA-positive for D. immitis and its Wolbachia were considered true positive for heartworm infection. Indeed, the presence of D. repens DNA or that of its Wolbachia as well as A. reconditum DNA indicates true positive infections. Results: The detection limit for Wolbachia and filariae qPCRs ranged from 5 × 10 −1 to 1.5 × 10 −4 mf/ml of blood. When tested on clinical samples, 29.2% (49/168) tested positive for filariae or Wolbachia DNA. Filarial species and Wolbachia genotypes were identified by the combined multiplex approach from all positive samples. Each species of Dirofilaria was significantly associated with a specific genotype of Wolbachia. Compared to the true positives, the approach showed excellent agreement (k = 0.98-1). Unlike D. immitis DNA, no A. vasorum DNA was detected by the duplex qPCR. The immunochromatographic test for heartworm antigen showed a substantial (k = 0.6) and a weak (k = 0.15) agreements before and after thermal pre-treatment of sera, respectively. Conclusions: The proposed approach is a reliable tool for the exploration and diagnosis of occult and non-occult canine filariosis. The current diagnosis of heartworm disease based on antigen detection should always be confirmed by qPCR essays. Sera heat pre-treatment is not effective and strongly discouraged.

The study reported in this paper was conducted to evaluate the digestibility of dietary carbohydrates (‘starch and sugars’ (S), neutral-detergent fibre (NDF), acid-detergent fibre (ADF)) and organic matter (OM) in the different parts of the equine digestive tract (stomach, jejuno-ileum, caecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, small colon and faeces). Three horses were given a standard diet (D1) based on fibrous pellets and straw and four were offered a high energy diet (D2) based on starch pellets and meadow hay. The digesta collection procedure, by total tract removal, permitted measurement of organ length, and dry matter and volume of digesta. Acid-detergent lignin (ADL) and acid insoluble ash (AIA) were used as natural digestibility markers. It was shown that AIA and ADL gave coherent data for ‘starch and sugars’ digestibility evaluation. ADL was a more relevant marker for parietal carbohydrates and OM digestibilities in horses given D1, whereas AIA have been preferred to evaluate these components digestibilities in horses offered D2. In horses given D1, precaecal OM digestibility coefficient varied from -0·04 to 0·20 whereas it varied from 0·46 to 0·62 in horses receiving D2. For both pellets, the results showed a considerable S digestibility occurring in the stomach (0·69 and 0·60 for D1 and D2 respectively) and this continued in the small intestine (0·88 and 0·89 for D1 and D2 respectively). With the exception of D2, structural carbohydrate fractions of the foods were poorly digested in the pre-caecal digestive parts. In the hindgut, OM digestibility coefficient varied from 0·47 to 0·60 for D1 and from 0·33 to 0·51 for D2. In horses given D1, highest digestibility was observed for each dietary carbohydrate in the left dorsal colon where it reached 0·99 for S; 0·45 for NDF and 0·40 for ADF. In horses receiving D2, the dietary components’ digestibilities increased regularly along the hindgut up to the faeces. The D2 structural fractions (NDF and ADF) digestibilities in the hindgut and faeces were lower than in horses given D1. These results not only confirmed that high energy diets like D2 can affect structural carbohydrate digestion in the horse hindgut but also indicated that a large amount of the energy part of the pelleted morning meal is broken down in the stomach.

Our knowledge of the microflora of the stomach of the horse is still limited, although some data indicate its important role in nutrition. The objective of this experiment was to investigate the microbial and biochemical profiles in the stomach of the horse and to quantify the disappearance of dietary starch. Total anaerobic bacteria, lactate-utilizing bacteria, lactobacilli, and streptococci were determined, and biochemical characteristics (pH, and DM, D- and L-lactate, D-glucose, NH3, and VFA concentrations) were measured in chyme collected from 4 horses by naso-gastric intubation aided by endoscopy, at 30 min before and 60, 120, and 210 min after the meal. The total anaerobic population exhibited a linear increase (5.54 to 6.98 log10 cfu/mL; P = 0.018) within the first postprandial hour and reached 8.32 log10 cfu/mL at 210 min after the meal. The concentrations of lactobacilli, streptococci, and lactate-utilizing bacteria in the stomach contents were 5.52, 4.82, and 6.95 log10 cfu/mL, respectively. Lactate concentration increased linearly from 0.25 mmol/L before the meal to 7.98 mmol/L at the last collection point (P = 0.013). This increase was mostly due to L-lactate accumulation. The VFA concentration increased linearly (P = 0.002) during the postprandial period from 1.96 to 8.17 mmol/L. Acetate represented, on average, 78 mol/100 mol of total VFA. The average concentration of NH3 in the stomach content was 2.48 mmol/L. Dietary starch disappearance did not respond during the post-prandial period and was not consistent with previous findings. These in vivo data provide complementary information on the postprandial microbial and biochemical kinetics in the stomachs of horses and confirm its abundant microbial colonization.