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The Salmonella enterica serovar Typhimurium znuABC genes encoding a high-affinity zinc uptake system and its regulatory zur gene have been cloned. Salmonella serovar Typhimurium zur and znuC knockout mutants have been constructed by marker exchange. The 50% lethal dose of the znuC mutant increased when either orally or intraperitoneally inoculated in BALB/c mice, while virulence of the zur mutant decreased only when mice were intraperitoneally challenged.

An experiment was designed to evaluate the effects of 3 different additives on the gastrointestinal microbiota of early-weaned pigs. Early-weaned (18 to 22 d; n = 32) pigs (6.0 +/- 0.10 kg of BW) from 8 litters were randomly distributed into 8 pens. Each pen was assigned 1 of 4 dietary treatments: a prestarter or control diet, the control diet with 0.04% avilamycin (AB), with 0.3% sodium butyrate, or with 0.03% plant extract mixture (XT; standardized mixture with 5% (wt/wt) carvacrol extracted from Origanum spp., 3% cinnamaldehyde extracted from Cinnamonum spp., and 2% capsicum oleoresin from Capsicum annum). At the end of the experimental period, 8 pigs per treatment were killed, and samples of their intestinal content were taken. The total bacterial load along the gastrointestinal tract (GIT; stomach, jejunum, cecum, and distal colon) and the lactobacilli and enterobacteria in the jejunum and cecum were measured by quantitative PCR. The total microbial counts along the GIT did not differ among the diets, but there was an increase in the lactobacilli:enterobacteria ratio in the cecum of the piglets on the XT diet (P = 0.003). Restriction fragment length polymorphism of the PCR-amplified V3, V4, and V5 regions of the 16S rDNA gene showed changes in the structure of the microbial community in the jejunum. Dendrograms grouped animals by diets; control with 0.3% sodium butyrate was the treatment that promoted the biggest changes in the microbial ecosystem, followed by AB and then XT. Biodiversity increased when using additives compared with the control diet (P = 0.002). Microbial metabolic activity along the hindgut was studied using the concentration of purine bases and carbohydrase activities. Different patterns for purine bases were observed between diets (diet x intestinal section, P = 0.01). The control diet reached a maximum purine base concentration at the end of the colon, whereas that of the AB diet was reached at the cecum. We could not detect any cellulase or xylanase activities in animals of this age. Appreciable amylase and amylopectinase activities were found, but they did not differ between diets. The results suggest that the effects of avilamycin, butyrate, or the plant extract would not be related to a reduction in the number of total bacteria inhabiting different sections of the GIT but rather to changes in the ecological structure and metabolic activity of the microbial community.

This chapter gives a general and brief description of the morphological and functional characteristics of the digestive system of the rabbit that may be important for understanding several digestive processes. The changes in these characteristics from the time of weaning until attainment of maturity are discussed.

The effect of neutral detergent-soluble fiber level on gut barrier function and intestinal microbiota was examined in weaned rabbits. A control diet (AH) containing 103 g of neutral detergent-soluble fiber/ kg of DM included alfalfa hay as main source of fiber. Another diet (B-AP) was formulated by replacing half of the alfalfa hay with a mixture of beet and apple pulp resulting in 131 g of soluble fiber/kg of DM. A third diet (OH) was obtained by substituting half of the alfalfa hay with a mix of oat hulls and a soybean protein concentrate and contained 79 g of soluble fiber/kg of DM. Rabbits weaned at 25 d and slaughtered at 35 d were used to determine ileal digestibility, jejunal morphology, sucrase activity, lamina propria lymphocytes, and intestinal microbiota. Suckling 35-d-old rabbits were used to assess mucosa morphology. Mortality (from weaning to 63 d of age) was also determined. Villous height of the jejunal mucosa increased with soluble fiber (P = 0.001). Rabbits fed with the greatest level of soluble fiber (BA-P diet) showed the highest villous height/ crypt depth ratio (8.14; P = 0.001), sucrase specific activity (8,671 mumol of glucose/g of protein; P = 0.019), and the greatest ileal starch digestibility (96.8%; P = 0.002). The opposite effects were observed in rabbits fed decreased levels of soluble fiber (AH and OH diets; 4.70, 5,848 mumol of glucose/g of protein, as average, respectively). The lowest ileal starch digestibility was detected for animals fed OH diet (93.2%). Suckling rabbits of the same age showed a lower villous height/crypt depth ratio (6.70) compared with the B-AP diet group, but this ratio was higher than the AH or OH diet groups. Lower levels of soluble fiber tended (P = 0.074) to increase the cellular immune response (CD8+ lymphocytes). Diet affected IL-2 production (CD25+, P = 0.029; CD5+CD25+, P = 0.057), with no clear relationship between soluble fiber and IL-2. The intestinal microbiota biodiversity was not affected by diets (P >/= 0.38). Rabbits fed the B-AP and AH diets had a reduced cecal frequency of detection compatible with Campylobacter spp. (20.3 vs. 37.8, P = 0.074), and Clostridium perfringens (4.3 vs. 17.6%, P = 0.047), compared with the OH diet group. Moreover, the mortality rates decreased from 14.4 (OH diet) to 5.1% (B-AP diet) with the increased presence of soluble fiber in the diet. In conclusion, increased levels of dietary soluble fiber improve mucosal integrity and functionality.

We evaluated spray-dried animal plasma (SDAP) as an alternative to antimicrobial medication with colistin sulfate in weanling pigs challenged with Escherichia coli K99. Forty-eight piglets weaned at 24 d of age were distributed into 12 pens, and each pen was assigned to one of four dietary treatments. All the piglets were given an oral dose of 5 × 10 7 cfu of E. coli K99 at weaning. The dietary treatments followed a factorial arrangement with two levels of SDAP (0 and 7%) and two levels of colistin (0 and 300 mg/kg of diet). The ADG and ADFI were measured on d 7 and 14 of trial. Three piglets from each treatment were killed on d 7 and 14 to remove the small intestine, and to obtain ileal and cecal digestive contents. The inclusion of SDAP improved ADG by 68 g (P < 0.05) and ADFI by 41 g (P < 0.10) in wk 1 of trial. During wk 2, SDAP improved ADG by 41 g (P < 0.10) and gain:feed ratio (G:F) by 25% (P < 0.01). On the other hand, whereas colistin had no effect on performance in wk 1, it improved ADG by 102 g (P < 0.01), ADFI by 62 g (P < 0.01), and G:F by 26% (P < 0.01) in wk 2. Over the 14 d of the trial, ADG was improved by 54 (P < 0.05) and

Avian pathogenic Escherichia coli (APEC) are the major cause of colibacillosis in poultry production. In this study, a total of 22 E. coli isolated from colibacillosis field cases and 10 avian faecal E. coli (AFEC) were analysed. All strains were characterised phenotypically by susceptibility testing and molecular typing methods such as pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). The presence of 29 virulence genes associated to APEC and human extraintestinal pathogenic E. coli (ExPEC) was also evaluated. For cephalosporin resistant isolates, cephalosporin resistance genes, plasmid location and replicon typing was assessed. Avian isolates belonged to 26 O:H serotypes and 24 sequence types. Out of 22 APEC isolates, 91% contained the virulence genes predictors of APEC; iutA, hlyF, iss, iroN and ompT. Of all strains, 34% were considered ExPEC. PFGE analysis demonstrated a high degree of genetic polymorphism. All strains were multi-resistant, including those isolated from healthy animals. Eleven strains were resistant to cephalosporins; six contained bla CTX-M-14, two bla SHV-12, two bla CMY-2 and one bla SHV-2. Two strains harboured qnrA, and two qnrA together with aac(6’)-Ib-cr. Additionally, the emergent clone O25b:H4-B2-ST131 was isolated from a healthy animal which harboured bla CMY-2 and qnrS genes. Cephalosporin resistant genes were mainly associated to the presence of IncK replicons. This study demonstrates a very diverse population of multi-drug resistant E. coli containing a high number of virulent genes. The E. coli population among broilers is a reservoir of resistance and virulence-associated genes that could be transmitted into the community through the food chain. More epidemiological studies are necessary to identify clonal groups and resistance mechanisms with potential relevance to public health.

The effect of dietary supplementation with 1% l-glutamine and a combination of 1% l-glutamine and 0.5% l-arginine on intestinal health was examined in weaned rabbits. A basal diet was formulated to meet nutrient recommendations. Another 2 diets were formulated by adding 1% (as-fed basis) Gln or a mixture of 1% (as-fed basis) Gln + 0.5% (as-fed basis) Arg (Gln-Arg) to the basal diet. In Exp. 1, a total of 357 rabbits were blocked by litter and assigned at random to the experimental diet to determine mortality (119 per diet) and growth performance (35 per diet; from weaning at 25 to 56 d of age). Rabbits were fed the experimental diets for a 2-wk period and thereafter received a commercial diet. Rabbits weaned at 25 d (blocked by litter and assigned at random to diets) were slaughtered at 35 d and used to determine apparent ileal digestibility of DM, CP, and AA (Exp. 2, a total of 60 rabbits), intestinal morphology, N-aminopeptidase and myeloperoxidase intestinal activity, the expression of PPARgamma at the ileum and kidney, serum immunoglobulin in healthy and sick rabbits (Exp. 3, a total of 24 rabbits), and ileal and cecal microbial composition by PCR-RFLP (Exp. 4, a total of 45 rabbits). Dietary treatment did not affect ADG, ADFI, or G:F, during the entire fattening period. Supplementation with Gln reduced mortality during the first 2 wk and the whole fattening period from 18.5 to 8.4% (P = 0.023) and from 31.9 to 20.2% (P = 0.039), respectively, whereas no effect was detected for Arg supplementation. Among all the variables studied, the reduction on mortality due to Gln was related to a reduced intestinal colonization (Eimeria lesions) and changes on microbial ecosystem in the ileum and cecum, reducing the frequency of detection of Clostridium spp. (from 86.7 to 33.3%, P = 0.003) at the ileum, and Helicobacter spp. at the ileum (from 86.7 to 46.7%, P = 0.003) and at the cecum (from 86.7 to 46.7, P = 0.028), whereas no effect was detected for Arg supplementation. In conclusion, 1% l-Gln supplementation to postweaned rabbit diets decreased fattening mortality and modified the intestinal microbiota (although no consistent effects were observed on mucosal histology or inflammatory and systemic immune response). Diets containing a combination of 1% Gln and 0.5% Arg were of little additional benefit.