It was hypothesized that dietary inclusion of Bacillus subtilis DSM 32315 could inhibit Clostridium perfringens induced necrotic enteritis (NE), thereby improving broiler performance. Male, d 0 chicks were randomly assigned 14 birds/pen, 11 pens/treatment in 3 treatments: a basal diet (control), a coccidiostat fed control (Narasin), and a direct fed microbial (DFM) B. subtilis DSM 32315 treatment. Necrotic enteritis was induced in all birds by oral inoculation of Eimeria maxima oocysts on d 12 and a virulent C. perfringens on d 16. Mortality was reduced ( P < 0.001) in DFM and Narasin compared to control. DFM reduced ( P < 0.001) feed conversion ratio (FCR) compared to control. Furthermore, DFM and Narasin reduced ( P < 0.001) footpad lesions. The DFM was shown to increase ( P < 0.05) Bacillus spp. and decrease ( P < 0.05) C. perfringens in the ileum and cecum at several time points. To investigate microbiome changes in the cecum, digesta samples were analyzed with % guanine and cytosine (%G+C) microbial profiling which fractionates bacterial chromosomes based on the %G+C in DNA. The method revealed treatment profile peaks in low (27.0 to 34.5%), mid (40.5 to 54.0%), and high (59.0 to 68.0%) G+C fractions. 16S rRNA gene amplification and high throughput sequencing was conducted on each of these fractions in order to elucidate specific bacterial population differences. In the low and mid %G+C fractions, DFM had greater abundance of Lactobacillaceae family members ( P = 0.03 and P = 0.01, respectively) and Lactobacillus salivarius ( P = 0.04 and P = 0.01, respectively) than control or Narasin. Lactobacillus johnsonii was also greater in the low %G+C fraction compared to control and Narasin ( P = 0.01). Lachnospiraceae ( P = 0.04) and Ruminococcaceae ( P < 0.01) in the mid %G+C fraction were reduced in the DFM compared to control. Positive alterations to the microbial populations in the gut of broilers may at least be a partial mechanism by which B. subtilis DSM 32315 reduced pathology and improved performance of broilers in the NE challenge.

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Our objective was to determine the effects of feeding oscillating compared with static dietary crude protein (CP) concentrations on nitrogen (N) retention and urea flux across ruminal epithelia. Twenty-seven Suffolk wether lambs (n = 9) were assigned to a medium-CP diet [MEDIUM; 127 g CP⋅kg dry matter (DM)(-1)] or to diets with oscillating CP content (OSC) fed in 2 different sequences, i.e. 2 d of low CP (103 g CP⋅kg DM(-1)) followed by 2 d of high CP (161 g CP⋅kg DM(-1); OSC-HIGH) or vice versa (OSC-LOW). Diet adaptation was for 24 d, followed by 8 d of total urine and feces collection. On d 33, lambs were slaughtered 4 h after the morning feeding, such that those receiving OSC-LOW and OSC-HIGH diets were slaughtered on d 3 of receiving the low- or high-CP diets, respectively. Ruminal epithelia were collected and mounted in Ussing chambers and the serosal-to-mucosal urea flux (J(sm-urea)) was measured using (14)C-urea. Ruminal NH(3)-N concentration was lower (P = 0.001) in lambs fed OSC-LOW compared with those fed OSC-HIGH. Although N intake was similar, retained N (P = 0.001) and microbial N supply (P = 0.001) were greater in lambs fed OSC compared with those fed MEDIUM. The total J(sm-urea) was higher (P = 0.001) in lambs fed OSC-LOW compared with those fed OSC-HIGH. Across diets, the addition of phloretin [a known specific inhibitor of facilitative urea transporter (UT)-B] reduced J(sm-urea) by 19.5-22.3% (P = 0.001); however, phloretin-insensitive J(sm-urea) was the predominant route for transepithelial urea transfer. Taken together, these data indicate that feeding oscillating dietary CP concentrations improves N retention partly by increasing urea recycling to the rumen when animals are fed low-CP diets, but the greater rates of urea transfer cannot be attributable to upregulation of UT-B.

The objective of this study was to evaluate the effects of the dietary supplementation of Bacillus subtilis DSM 32315 (probiotic) on the performance and intestinal microbiota of broiler chickens infected with Clostridium perfringens (CP). One-day-old broiler chickens were assigned to 3 treatments with 8 replicate pens (50 birds/pen). The treatments were: non-infected control; infected control; and infected supplemented with probiotic (1 × 106 CFU/g of feed). On day of hatch, all birds were sprayed with a coccidia vaccine based on the manufacturer recommended dosage. On d 18–20 the infected birds were inoculated with CP via feed. Necrotic enteritis (NE) lesion score was performed on d 21. Digestive tract of 2 birds/pen was collected on d 21 to analyze the ileal and cecal microbiota by 16S rRNA sequencing. Performance was evaluated on d 28 and 42. On d 21, probiotic supplementation reduced (p < 0.001) the severity of NE related lesion versus infected control birds. On d 28, feed efficiency was improved (p < 0.001) in birds supplemented with probiotic versus infected control birds. On d 42, body weight gain (BW gain) and feed conversion ratio (FCR) were improved (p < 0.001) in probiotic supplemented birds versus infected control birds. The diversity, composition and predictive function of the intestinal microbial digesta changed with the infection but the supplementation of probiotic reduced these variations. Therefore, dietary supplementation of Bacillus subtilis DSM 32315 was beneficial in attenuating the negative effects of CP challenge on the performance and intestinal microbiota of broilers chickens.

A study was conducted to investigate the effects of Bacillus amyloliquefaciens CECT 5940 as a direct-fed microbial (DFM) alone or in association with bacitracin methylene disalicylate (BMD) in broilers under enteric pathogen challenge. A total of 1,530-day-old male Cobb500 chicks were randomly assigned to 5 treatments, with 9 replicate pens with 34 birds each. Treatments included positive control (PC, basal diet without additives or challenge); negative control (NC, basal diet without additive and challenged birds); NC + 0.05 g/kg BMD; NC + 1 g/kg DFM (106 CFU B. amyloliquefaciens CECT 5940/g of feed); and NC + 0.05 g/kg BMD + 1 g/kg DFM. The challenge consisted of oral gavage with Eimeria maxima and Clostridium perfringens inoculum. Body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) were evaluated on days 21, 35, and 42. Ileal and cecal content were collected on days 21 and 28 for C. perfringens enumeration by real-time PCR assay and the intestinal health was evaluated by scores. Uniformity (UN), carcass (CY), and breast meat yields (BMY) were evaluated on day 42. After 14 and 21 d post-inoculation, birds in the challenged groups had significant lower FI and BWG compared to the PC group (P < 0.05). However, the groups receiving DFM, BMD, or its combination presented better FCR, CY, BMY, UN, and lower incidence of footpad lesion and litter quality visual scores, compared to the NC group without feed additives (P < 0.05). Mortality was not affected by treatments (P > 0.05). Broilers fed DFM, BMD, or its combination presented lower C. perfringens in ileal content at 21 and 28 d compared to NC group without additives (P < 0.05) and also maintained gut health by keeping the frequency of ballooning, abnormal content, and swollen mucosa comparable to the PC group (P > 0.05). The study indicates that Bacillus amyloliquefaciens CECT 5940 is effective as BMD to provide similar performance and gut health in challenged broilers.