The effects of the supplementation of Bacillus subtilis RX7 and B2A strains on the performance,blood profiles,intestinal Salmonella concentration,noxious gas emission,organ weight and breast meat quality of broiler challenged with Salmonella typhimurium

An experiment was conducted to evaluate the effects of B. subtilis RX7 and B. subtilis B2A on growth performance, blood profiles, intestinal Salmonella population, noxious gas emission, organ weight and breast meat quality of broilers under S. typhimurium challenge. A total of 120, one‐day‐old Ross 308 male broiler chicks were assigned to four dietary treatments, composed of six replications, with five birds per replication, for 10 day. The dietary treatment groups were negative control (NC; no antibiotic, no B. subtilis), positive control (PC; NC + 0.1% virginiamycin), B. subtilis RX7 (NC + 0.1% B. subtilis RX7 1.0 × 10⁹ cfu/g) and B. subtilis B2A (NC + 0.1% B. subtilis 1.0 × 10⁹ cfu/g). All birds were orally challenged with 2 ml suspension, containing 10⁴ cfu/ml of S. typhimurium KCCM 40253. Results indicated that the body weight gain, feed intake and feed conversion did not differ, among all comparative treatments. Serum haptoglobin concentration was lower in Bacillus treatments (RX7 + B2A) than the NC treatment (p < 0.05). Intestinal and excreta Salmonella number, and excreta ammonia gas emission in the PC treatment or Bacillus treatments, was lower than the NC treatment (p < 0.05). Breast pH, colour and water‐holding capacity were not affected by supplementation of B. subtilis RX7 and B2A. However, drip loss at 1 day post‐slaughter from birds fed with B. subtilis RX7 and B2A decreased, compared with the positive control birds (p < 0.05). Relative gizzard weights of birds fed B. subtilis RX7 and B2A were significantly higher than the NC birds under S. typhimurium challenge. It is concluded from the results that B. subtilis RX7 and B2A increased the gizzard weight and decreased the intestinal and excreta Salmonella population and excreta ammonia gas, and drip loss of breast meat after being stored for 1 day, under stress caused by the S. typhimurium challenge.     

Effects of composite antimicrobial peptide on growth performance and health in weaned piglets

This study was conducted to evaluate the effects of composite antimicrobial peptide (CAP) on growth performance and health status in weaned piglets. Over 28 days, 36 weaned piglets (body weight, 10.58 ± 0.99 kg) underwent three treatments: negative control (NC, basal diet), positive control (PC, basal diet + 20 mg/kg colistin sulphate + 50 mg/kg kitasamycin), and CAP treatment (CAP, basal diet with 400 mg/kg CAP). Average daily gain of piglets fed the CAP diet was greater (P < 0.05) than that of piglets fed the PC or NC diet during days 1–7, 8–14 and 15–21. Diarrhea rates of piglets fed the CAP or PC diet were lower (P < 0.05) than those of NC‐fed piglets during days 1–7. Apparent total tract digestibility for dry matter and crude ash in CAP‐fed piglets was greater (P < 0.05) than that of NC‐fed piglets. In the CAP group, Lactobacillus and Bifidobacterium counts were greater (P < 0.05) and Escherichia coli counts were lower (P < 0.05) than numbers for the NC group. Our results indicate that dietary CAP had beneficial effects on growth performance and health status in weaned piglets.     

Positive effects of dietary supplementation of three probiotics on milk yield,milk composition and intestinal flora in Sannan dairy goats varied in kind of probiotics

In this study, we investigated the effects of Saccharomyces cerevisiae (SC), Bacillus subtilis (BS) and Enterococcus faecalis (EF), singly and in combination, on the dry matter intake (DMI), milk production and composition, and faecal microflora of Saanen dairy goats. Fifty goats were randomly divided into five groups: (a) basal diet (control); (b) basal diet + SC; (c) basal diet + BS; (d) basal diet + EF; and (e) basal diet + mixed probiotics. Each treated animal received 5 g/d of probiotics for a total administration of 5 × 1,011 CFU/goat per day. The inclusion of B. subtilis and E. faecalis in the diet of lactating Saanen goats increased DMI (p < .05). Enhanced milk yield was observed with BS and EF. Milk fat percentage was significantly increased by feeding mixed probiotics compared with the control (p < .05); supplying SC, BS and mixed probiotics enhanced the protein percentage (p < .05). The milk lactose percentage in the SC and BS groups was higher than in the control (p < .05). The amount of milk total solids was higher after feeding EF or mixed probiotics than in the control group (p < .05). Non-fat solids showed no notable differences among groups (p > .05). There was no significant influence on gut bacterial abundance and diversity from adding these three probiotics, singly or in combination. Bacteroidales, Escherichia–Shigella and Christensenellaceae abundances were decreased by supplying these probiotics but Succinivibrionaceae increased. In conclusion, there were positive influences of probiotic feed supplementation on intake, milk performance and intestinal microecology.     

Effect of protease derived from Pseudoalteromonas arctica supplementation on growth performance,nutrient digestibility,meat quality,noxious gas emission and blood profiles in finishing pigs

This study was conducted to evaluate the effects of dietary supplementation of protease derived from Pseudoalteromonas arctica (PPA) in finishing pigs. A total of 160 pigs were used in this 10‐week trial. Dietary treatment groups were as follows: CON (basal diet); TRT1 (basal diet + 0.1% PPA); TRT2 (basal diet + 0.2% PPA); and TRT3 (basal diet + 0.3% PPA). During weeks 1–5, pigs fed with different levels of PPA‐supplemented diet showed linear increase (p < .05) in the apparent total tract digestibility (ATTD) of nitrogen (N) and linear decrease (p < .05) in the concentrations of serum total protein. During weeks 6–10, pigs fed with different levels of PPA‐supplemented diet showed a linear decrease in feed conversion ratio (p < .05). During the overall period, there was a linear decrease in feed conversion ratio (p < .05) associated with the inclusion of PPA. Pigs fed diets with 0.2% PPA supplementation had lower (p < .05) feed conversion ratio than those fed CON diet during weeks 6–10 and the overall period, and had higher (p < .05) ATTD of N than those fed CON diet during weeks 1–5. Pigs fed diets with PPA supplementation had lower (p < .05) concentrations of serum total protein than those fed CON diet on week 5. In conclusion, dietary supplementation with PPA diet has beneficial effects on growth performance, nutrient digestibility, backfat thickness and the concentrations of serum total protein.     

Effects of Bacillus subtilis C‐3102 on production,hatching performance,egg quality,serum antioxidant capacity and immune response of laying breeders

To investigate the supplemental effects of Bacillus subtilis C‐3102 on the production, hatching performance, egg quality, serum antioxidant capacity and immune response of laying breeders, a total of 480 Xuefeng black‐bone (25‐week‐old) hens were randomly assigned into four treatment groups: Hens fed the basal diets with 0 (CON), 3.0 × 10⁵ (BS‐1), 6.0 × 10⁵ cfu/g (BS‐2) and 9.0 × 10⁵ (BS‐3) cfu/g of B. subtilis C‐3102. As the B. subtilis C‐3102 level increased, egg weight (linear, p < 0.01; quadratic, p = 0.003), fertility (linear, p = 0.021; quadratic, p = 0.059), hatchability (linear, p = 0.038; quadratic, p = 0.119) and yolk colour (linear, p = 0.006; quadratic, p = 0.021) increased in a linear or quadratic manner. Yolk index increased quadratically (linear, p = 0.054; quadratic, p = 0.017), and eggshell thickness (linear, p = 0.036; quadratic, p = 0.128), the activity of GSH‐Px (linear, p = 0.024; quadratic, p = 0.078), the concentration of IgM (linear, p = 0.016; quadratic, p = 0.056) and the level of AIV‐Ab (linear, p = 0.034; quadratic, p = 0.103) in the serum increased linearly as dietary supplementation of B. subtilis C‐3102 increased. The results showed that dietary treatments did not affect egg production, feed conversion ratio, egg mass, hatchability of fertile eggs, eggshell‐breaking strength, egg‐shape index, yolk percentage, Haugh unit, T‐SOD, T‐AOC, MDA, IgA and IgG concentrations and the level of NDV‐Ab in the serum. In conclusion, dietary supplementation of 9.0 × 10⁵ cfu/g B. subtilis C‐3102 in laying breeders diets may be a feasible means of effectively increasing egg weight, fertility and hatchability, and improving egg quality such as eggshell thickness, yolk index and yolk colour. Besides, B. subtilis C‐3102 can enhance the activity of GSH‐Px, the concentration of IgM and the level of AIV‐Ab in the serum.     

Medium‐chain glycerides affect gut morphology,immune‐ and goblet cells in post‐weaning piglets: In vitro fatty acid screening with Escherichia coli and in vivo consolidation with LPS challenge

The influence of medium‐chain glycerides on performance and gastrointestinal well‐being in weaning piglets was assessed. First, caproic (C6), caprylic (C8) and capric (C10) acid activity against Escherichia coli was screened in vitro. Pig flora of the whole small intestine was used as inoculum. Seven in vitro incubations were done in duplicate at pH = 3 and 5: C10 (15 mM), C8 (12 mM), C6 (15, 12, 10 mM), a non‐incubated‐negative control and incubated negative control. Culture suspensions were plated on E. coli‐selective agar. Controls showed bacterial growth. C6 and C8 showed no growth at both pH‐values, where C10 showed growth at pH = 5. Secondly, an in vivo study was done with 80 weaned piglets over 42 days, housed in pens of eight animals (five pens/treatment), fed a basal diet containing broken rice/soya bean meal/fish meal and supplemented with C6 and C8 in medium‐chain glyceride form (MCT6/8, 0.175%) or antibiotic growth promoter (AGP, 0.020%) (Kasetsart University, Thailand) serving as control. Feed intake, daily gain and feed‐to‐gain ratio did not differ between MCT6/8 and AGP. Per replicate, two random selected piglets were challenged intravenously with E. coli‐lipopolysaccharide (LPS) or saline solution (S) at Days 21 and 28. All challenged animals were sacrificed; blood and digestive tract samples (jejunum/ileum) were collected at Day 35. LPS challenge consistently reduced villus height and crypt depth for MCT6/8 and AGP. However, LPS‐challenged piglets supplemented with MCT6/8 restored villus height, where AGP did not. MCT6/8 piglets had higher serum IgA, more jejunal IgA‐positive plasma cells and goblet cells than AGP. At the ileal level, results were similar, though less pronounced. The present study offers new insight in the benefits of MCT6/8 over AGP in the post‐weaning period. There is in vitro anti‐microbial action of C6 and C8 on E. coli. In vivo, MCT6/8 also has protective effects in the small intestine that may result in growth promotion.     

Influence of Bacillus subtilis GCB‐13‐001 on growth performance,nutrient digestibility,blood characteristics,faecal microbiota and faecal score in weanling pigs

The present study investigated the influence of Bacillus subtilis GCB‐13‐001 on growth performance, nutrient digestibility, blood characteristics, faecal microbiota and faecal score in weanling pigs. A total of 120 weaning pigs [(Landrace × Yorkshire) × Duroc; 7.73 ± 0.75 kg (28 days of age)] were randomly allotted into three treatments according to their initial body weight (BW) and gender in a 6‐week experiment. There were 8 replication pens in each treatment, with five pigs/pen. Dietary treatment groups were as follows: (a) basal diet (CON), (b) CON + 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁸ CFU/kg (T1) and (c) CON + 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁹ CFU/kg (T2). Days 1 to 7, the BW and ADG with T2 treatment were higher (p < .05) than CON treatment, as well as F:G showed trends in linear reduction (p < .1). Days 8 to 21, the BW and ADG were improved (p < .05) in pigs offered T1 and T2 diets compared with CON diet. Days 22 to 42, BW and ADG were higher (p < .05) in pigs fed T2 diet than CON and T1 diets, and the pigs fed T1 diet had higher BW than CON treatment. Overall, the ADG with the T2 treatment was higher (p < .05) than that with the T1 and CON treatments, and pigs offered T1 treatment had higher (p < .05) ADG than CON treatment. Moreover, F:G ratio were significantly decreased (p < .05) by T2 treatment compared with CON treatment. The faecal Lactobacillus counts were improved, and E. coli counts were reduced (p < .05) in pigs fed T2 diet compared with CON at the end of the experiment. In conclusion, supplementation of 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁹ CFU/kg has shown a beneficial effect in improving BW, increase ADG, decrease F:G ratio.     

β‐hydroxy‐β‐methyl butyrate promotes leucine metabolism and improves muscle fibre composition in growing pigs

The aim of this study was to investigate the effects of excess leucine (Leu) vs. its metabolites α‐ketoisocaproate (KIC) and β‐hydroxy‐β‐methyl butyrate (HMB) on Leu metabolism, muscle fibre composition and muscle growth in growing pigs. Thirty‐two pigs with a similar initial weight (9.55 ± 0.19 kg) were fed 1 of 4 diets for 45 days: basal diet, basal diet + 1.25% L‐Leu, basal diet + 1.25% KIC‐Ca, basal diet + 0.62% HMB‐Ca. Results indicated that relative to the basal diet and HMB groups, Leu and KIC groups exhibited increased Leu concentrations and decreased concentrations of isoleucine, valine and EAAs in selected muscle (p < 0.05) and had lower mRNA levels of MyHC I and higher expression of MyHC IIx/IIb (p < 0.05), and there was no significant difference between the basal and HMB‐supplemented groups. Moreover, the mRNA expression levels of AMPKα and UCP3 were higher but the myostatin mRNA levels were lower in the soleus muscle of the HMB group than those from other groups (p < 0.05). These findings demonstrated that doubling dietary Leu content exerted growth‐depressing effects in growing pigs; dietary KIC supplementation induced muscular branched‐chain amino acid imbalance and promoted muscle toward a more glycolytic phenotype; while dietary HMB supplementation promoted the generation of more oxidative muscle types and increased muscle growth specially in oxidative skeletal muscle, and these effects of HMB might be associated with the AMPKα‐Sirt1‐PGC‐1α axis and mitochondrial biogenesis.     

The Effect of Lactobacillus isolates on growth performance,immune response,intestinal bacterial community composition of growing Rex Rabbits

The objective of this experiment was to investigate the effect of Lactobacillus isolates on intestinal bacterial community composition of growing Rex Rabbits. A total of 120 weaned Rex Rabbits (30 days old, 30 per group) were used for the experiment, which started after an adaptation period of 7 days. The control group was fed with basal diet only, while the treatment I, II and III groups were fed with basal diet adding antibiotics, Lactobacillus zeae (LB1) and Lactobacillus casei (L3) respectively. Growth performance, immune response and intestinal flora have been examined. The results obtained were as follows: (i) F/G of the rabbits fed with Lactobacillus isolates was significantly lower than that of the control group (p < 0.05). (ii) The concentration of ALT decreased significantly (p < 0.05) and that of IgG and IgM increased significantly (p < 0.05) after feeding rabbits with Lactobacillus isolates. (iii) Lactobacillus isolates had no influence on the number of mast cells in duodenum and jejunum, but increased the number of mast cells in caecum significantly (p < 0.05). (iv) The data from pyrosequencing‐based analysis suggested that the bacterial community in the rabbit's intestinal flora can be changed by Lactobacillus isolates and antibiotics, especially for the microbial diversity and abundance in the caecum. In conclusion, the application of proper Lactobacillus isolates can improve the growth performance, enhance the immunological function and adjust the intestinal micro‐ecosystem of growing Rex Rabbits.     

Vaginal isolation of beta‐haemolytic Streptococcus from bitches with and without neonatal deaths in the litters

The aim of the study was to identify beta‐haemolytic streptococci in the vagina of bitches who had delivered healthy litters and bitches who had delivered litters in which neonatal deaths occurred. Fifty‐one bitches divided into two groups were used. Group 1 (G1) included 28 bitches that had delivered healthy litters and group 2 (G2) included 23 bitches that had delivered puppies who died in the neonatal period. Two vaginal samples were taken, one in proestrus and the other at the end of gestation (EG). Beta‐haemolytic Streptococcus (BS) was isolated from 16 bitches (57%) in G1 and from 21 bitches (91%) in G2. The bacteriological cultures, serological tests (Streptex^®) and PCR assay allowed identification of Streptococcus canis and Streptococcus dysgalactiae in G1 and G2. Ultramicroscopic studies allowed the observation of M Protein and capsules in strains of S. dysgalactiae and S. canis in G1 and G2. The S. canis strains isolated from G2 showed thicker capsules than S. canis strains isolated from G1 (234 ± 24.2 vs 151.23 ± 28.93 nm; p < .001.). No differences were observed in capsule thickness between strains of S. dysgalactiae isolated from G1 and G2 (210 ± 13.54 vs 211.66 ± 19.67 nm; p > .70). All strains of beta‐haemolytic Streptococcus isolated were penicillin sensitive. Penicillin was administered from EG to 5 days post‐partum in 10 G2 females with isolation of BS (G2A). Saline solution was administered in eleven G2 females with isolation of BS (G2B). Ninety per cent of the puppies survived in G2A and 25% survived in G2B. Our results suggest BS is involved in canine neonatal deaths.     

Effect of oral administration of probiotics on growth performance,apparent nutrient digestibility and stress‐related indicators in Holstein calves

This study aimed to investigate the effect of dietary supplementation with Lactobacillus plantarum and Bacillus subtilis on growth performance, apparent nutrient digestibility and stress‐related indicators in dairy calves. Twenty‐four neonatal Holstein calves were randomly allocated to three treatments: a basal diet with no supplementation (control), the basal diet supplemented with 1.7 × 10¹⁰ CFU per head per day (CFU/h.d) of L. plantarum GF103 (LB group) or the basal diet supplemented with a mixture of L. plantarum GF103 (1.7 × 10¹⁰ CFU/h.d) and B. subtilis B27 (1.7 × 10⁸ CFU/h.d) (LBS group). Dry matter intake (DMI), average daily gain (ADG), feed conversation ratio (FCR), apparent digestibility of nutrients and stress‐related indicators were measured in this trail. The result indicated that no significant differences were observed in DMI or ADG (p > 0.05), but the FCR was improved in the LB group over the first 12 weeks (p > 0.05). The apparent digestibility of nutrients was not altered by probiotics in week 6 (p > 0.05), but the apparent digestibility of total phosphorus was significantly greater in the LB and LBS groups in week 8 (p > 0.05); additionally, an increase in the apparent digestibility of crude protein was detected in the LBS group (p > 0.05). Oral administration of L. plantarum alone improved the T‐lymphocyte transformation rate on days 58 and 62 (p > 0.05), while adding the mixture of L. plantarum and B. subtilis increased the T‐lymphocyte transformation rate (p > 0.05) but decreased the content of cortisol on day 58 (p > 0.05). No significant differences were detected between the LB and LBS groups in growth performance, apparent digestibility of nutrients and stress‐related indicators (p > 0.05). The results suggested that oral administration of L. plantarum improved growth performance, nutrient digestibility and relieved weaning stress in calves, but no additional effect was obtained by supplementation with B. subtilis.     

The effects of dietary supplementation with porous zinc oxide on growth performance,intestinal microbiota,morphology, and permeability in weaned piglets

The objective of this experiment was to evaluate the effects of dietary supplementation with porous zinc oxide (HiZox) on growth performance, intestinal microbiota, morphology, and permeability in weaned piglets. A total of 128 weaned piglets [(Landrace × Yorkshire) × Duroc] with an average body weight (BW) of (6.55 ± 0.25 kg; 21 d of age) were randomly assigned to four dietary treatments: (1) a corn‐soybean basal diet; (2) basal diet + 3,000 mg/kg conventional ZnO; (3) basal diet + 200 mg/kg HiZox; (4) basal diet + 500 mg/kg HiZox. The experiments lasted for 28 days. Incremental HiZox in the diet increased ADG (linear p = 0.015; quadratic p = 0.043) and ADFI (linear p = 0.027; quadratic p = 0.038), and the diarrhea index decreased linearly and quadratically (p < 0.01) as HiZox supplemented increased. Furthermore, supplementation with HiZox increased the amounts of Lactobacillus spp. (p < 0.05) in the ileum and cecum in comparison with that of control treatment or 3,000 mg/kg ZnO treatment, while decreased the populations of Escherichia coli, Clostridium coccoides, and Clostridium. leptum subgroup (p < 0.05) in the ileum and cecum relative to those in control treatment. The addition of HiZox increased the villus height and villus‐to‐crypt ratio (VC) of duodenum, jejunum, and ileum (p < 0.05), while decreased the crypt depth of jejunum (p < 0.05) and tended to reduce the crypt depth of duodenum (p < 0.10) compared with the control treatment. Piglets fed with 500 mg/kg HiZox had lower serum D‐lactate and diamine oxidase (DAO) than those fed with basal control diet or 3,000 mg/kg ZnO diet (p < 0.01). The results suggested that supplementation with HiZox modulated intestinal microbial composition and improved intestinal morphology, which may exert protective effects on the integrity of the mucosal barrier function of weaned piglets, was as efficacious as pharmaceutical doses of ZnO in enhancing growth performance, indicating that the HiZox may be a promising alternative to pharmaceutical doses of ZnO.     

Differential responses of weaned piglets to supplemental porcine or chicken plasma in diets without inclusion of antibiotics and zinc oxide

This study was conducted to investigate the effects of spray-dried porcine plasma protein (SDPP) or spray-dried chicken plasma protein (SDCP) supplementation in diets without the inclusion of antibiotics and zinc oxide (ZnO) on growth performance, fecal score, and fecal microbiota in early-weaned piglets. A total of 192 healthy weaning piglets (Duroc × Landrace × Yorkshire, 21 d old) were blocked by BW (6.53 ± 0.60 kg) and randomly assigned to 4 dietary treatments: negative control (NC, basal diet), positive control (PC), basal diet + ZnO at 2 g/kg and antibiotics at 0.8 g/kg), SDPP (containing 5% SDPP), and SDCP (containing 5% SDCP). The experiment lasted 14 d. The SDPP group had higher (P < 0.05) final BW, average daily gain and average daily feed intake than the NC and SDCP groups. The percentage of piglets with fecal scores at 2 or ≥2 was higher (P < 0.05) in the NC and SDCP groups than in the PC group. A decreased (P < 0.05) bacterial alpha diversity and Bacteroidetes abundance, but increased (P < 0.05) Firmicutes abundance were observed in the PC and SDPP groups when compared to the NC group. The relative abundance of Lactobacillus was higher (P < 0.05) in the SDPP than in the SDCP group, and that of Streptococcus was higher (P < 0.01) in the PC and SDPP groups than in the NC group. The PC group also had higher (P < 0.01) Faecalibacterium abundance than the NC and SDCP groups. Additionally, the SDCP group had higher (P < 0.05) serum urea nitrogen than those fed other diets, and lower (P < 0.10) short-chain fatty acids to branched-chain fatty acids ratio than the PC and SDPP groups. Overall, SDPP was a promising animal protein for piglets in increasing feed intake, modifying gut microbiota profile, reducing gut protein fermentation and alleviating diarrhea frequency, thus promoting growth performance, under the conditions with limited in-feed utilization of antibiotics and ZnO.     

Optimization of surfactin production from Bacillus subtilis in fermentation and its effects on Clostridium perfringens‐induced necrotic enteritis and growth performance in broilers

Bacillus species are commonly used as probiotics in the poultry feed industry for preventing infectious diseases and improving productivity by altering gastrointestinal microbiota. The growth parameters of Bacillus subtilis for surfactin production in fermentation and the benefits of surfactin on broiler chickens remain unclear. In this study, we examined the growth parameters of B. subtilis in fermentation and evaluated the effects of surfactin from B. subtilis‐fermented products on Clostridium perfringens‐induced necrotic enteritis and growth performance in broilers. Results showed that the highest viable biomass of B. subtilis was observed at 10% molasses and 2% yeast supplementation during fermentation. The 4‐ and 6‐day fermented B. subtilis products were heat‐, acid‐ and bile‐resistant. Furthermore, the 4‐day fermented B. subtilis products with the highest surfactin concentration showed the maximal antimicrobial activity against pathogens, including Escherichia coli, Staphylococcus aureus, Salmonella typhimurium and C. perfringens. Dietary B. subtilis‐fermented product supplementation in broilers significantly improved intestinal morphology and necrotic lesions under C. perfringens challenge. Bacillus subtilis treatments could enhance broiler productivity, as well as promote bone quality and intestinal morphology. These results together indicate that B. subtilis‐fermented products containing surfactin have potential for the development as feed additives and use as possible substitutes for antibiotics to treat C. perfringens in the poultry industry.     

Effect ofBacillus amyloliquefaciens-based direct-fed microbials and antibiotic on performance,nutrient digestibility,cecal microflora,and intestinal morphology in broiler chickens

Two hundred eighty-eight 1-d-old male Arbor Acres broilers were randomly allocated to 4 treatments to examine the effect of usingBacillus amyloliquefaciens-based direct-fed microbials (DFM) to replace antibiotics in the late growth stage on performance, nutrient digestibility, cecal microflora, and intestinal morphology in broilers. The experimental diets were (1) a negative control (NC) group fed a basal diet without any antibiotics; (2) a positive control (PC) group fed the basal diet with 200 mg/kg of zinc bacitracin; (3) a test group fed the basal diet with 200 mg/kg of zinc bacitracin from d 1 to 21 and 30 mg/kg of DFM from d 22 to 42 (DFM30); and (4) a test group fed the basal diet with 200 mg/kg of zinc bacitracin from d 1 to 21 and 60 mg/kg of DFM from d 22 to 42 (DFM60). The results showed that DFM30 and DFM60 increased BW gain and improved FCR as compared with NC group (P < 0.05). Generally, the PC, DFM30, and DFM60 groups showed increased apparent total tract nutrient digestibility for DM, CP, and AME as compared with the NC group. Both the DFM30 and DFM60 groups decreased cecalEscherichia coli population as compared with the NC group, and increased the population ofLactobacillus as compared with the PC group. The PC, DFM30, and DFM60 groups increased villus height and the ratio of villus height to crypt depth compared with NC. In conclusion, dietary supplementation withB. amyloliquefaciens-based DFM during d 22 to 42 may enhance performance by increasing nutrient digestibility and improving intestinal health in birds fed diets supplemented with zinc bacitracin from d 1 to 21.     

Inclusion of Bacillus amyloliquefaciens strain TOA5001 in the diet of broilers suppresses the symptoms of coccidiosis by modulating intestinal microbiota

Coccidiosis is an intestinal parasitic infection and one of the most prevalent and economically damaging diseases of chickens. Furthermore, coccidia‐induced mucogenesis promotes secondary colonization by Clostridium perfringens, a major pathogen of chickens that causes necrotic enteritis. Our previous work found that supernatant of a culture of Bacillus amyloliquefaciens strain TOA5001 (BA) inhibited the growth of C. perfringens on Gifu anaerobic broth medium. Accordingly, we evaluated the effectiveness of dietary BA administration in inhibiting C. perfringens colonization of the intestine in broilers that were experimentally infected with coccidia. Ten healthy broilers from a BA‐supplemented (2 × 10⁵ colony‐forming units/g of feed) broiler group and 10 from a non‐treated group were challenged with Eimeria tenella and E. maxima (5000 oocysts of each species/chick) at 28 days old. At 36 days old, five chicks from each group were slaughtered, whereas the remaining five in each group were killed at 49 days old. Dietary BA administration into Eimeria‐challenged birds reduced coccidial symptoms such as intestinal lesions. It also modified the cecal microbiota through suppressing C. perfringens and E. coli colonization, and inducing domination of Faecalibacterium prausnitzii, the Lactobacillus group and unknown Lachnospiraceae genera by bacterial DNA‐based metagenome analyses. B. amyloliquefaciens TOA5001 supplementation suppressed the symptoms of coccidiosis by modulating cecal microbiota in Eimeria‐challenged broilers.     

Modulation of broilers’ caecal microflora and metabolites in response to a potential probiotic Bacillus amyloliquefaciens

Studies have found that a dietary supplement of Bacillus amyloliquefaciens improved the growth performance, increased the nutrient digestibility of hosts and modulated the intestinal microflora. A total of 360 1‐day‐old Ross broilers were randomly divided into three treatments: a control group with a basal diet, an antibiotic group with a basal diet and added colistin sulphate, and a probiotics group with a basal diet and added Bacillus amyloliquefaciens. The HiSeq high‐throughput sequencing analysis of 16S rRNA was used to investigate the differences in birds’ caecal microflora, and metabolomics was used to analyse changes in caecal metabolites. Results showed that the supplementation of Bacillus amyloliquefaciens significantly improved the BW and ADG compared with the control birds. Results of sequencing indicated that (i) 645, 670, 596 unique operational taxonomic units (OTUs) were found in birds supplemented with Bacillus amyloliquefaciens on day 7, 21 and 42, separately, (ii) due to the diversity and relative abundance of the birds’ caecal microflora, the OTUs of the caecal microflora clustered according to age and treatment, except on day 42, (iii) among the six predominate families (Ruminococcaceae, Lachnospiraceae, Enterobacteriaceae, Erysipelotrichaceae, Lactobacillaceae and Rikenellaceae), the supplementation of Bacillus amyloliquefaciens significantly increased Enterobacteriaceae on day 42, (iv) Bacillus amyloliquefaciens increased the relative abundance of Faecalibacterium and Ruminococcus on day 21, increased the Faecalibacterium and Blautia and decreased the Ruminococcus on day 42. The metabolomics of caecal metabolites showed that the dietary Bacillus amyloliquefaciens changed the caecal metabolites involved of amino acid metabolism and glyceride metabolism, and the antibiotics changed the caecal metabolites that were related to carbohydrates and amino acid metabolism on day 21.     

Effects of Antimicrobial peptides on egg production,egg quality and caecal microbiota of hens during the late laying period

This study was conducted to determine the effects of diet supplementation of laying hens with antimicrobial peptides (AMP) on egg production, egg quality and caecal microbiota. A total of 360 Hy‐Line Brown laying hens (72 weeks old) were divided into three groups with four replicates of 30 birds each. The laying hens were fed with the basal diet (Control), the basal diet + 50 mg/kg AMP (group 1) and the basal diet + 100 mg/kg AMP (group 2). The experiment lasted for 45 d. Eggs were collected daily and caecal samples were collected at the end of the experiment. The results showed that AMP supplementation caused a significantly increased laying rate and decreased feed/egg ratio (p ＜ .05). Meanwhile, a distinctive difference in cecal microbiota was observed between AMP and control groups and the average values of microbial diversity and richness were lower in the AMP group than in the control group. At the phylum level, the relative abundance of Verrucomicrobia and Cyanobacteria were lower in the AMP group than in the control group. In conclusion, the results indicated that dietary supplementation with AMP can improve egg production and affect the cecal microbial community membership and structure of hens during late laying period.     

Effects of lactulose supplementation on performance,blood profiles,excreta microbial shedding of Lactobacillus and Escherichia coli,relative organ weight and excreta noxious gas contents in broilers

This study was to evaluate the effects of lactulose supplementation on performance, blood profiles, excreta microbial shedding of Lactobacillus and Escherichia coli, relative organ weight and excreta noxious gas contents in broilers. A total of 720 ROSS 308 broilers with a body weight of 46 ± 0.1 g (1 day of age) were used in a 28‐d experiment. Broilers were randomly allotted to 4 experiment diets with 12 replicate pens and 15 birds per pen. Dietary treatments were as follows: NC, negative control (without antibiotic); PC, NC + 0.1% tiamulin; L1, NC + 0.1% lactulose; and L2, NC + 0.2% lactulose. Broilers were fed with phase 1 (1–8 day), phase 2 (9–18 day) and phase 3 (19–28 day) diets in the form of mash. During day 1–8, broilers fed the PC and L2 diets had higher (p < 0.05) body weight gain than those fed the NC diet. During day 19–28, broilers fed the L1 and L2 diets had lower (p < 0.05) feed intake than those fed the NC diet. The feed conversion ratio (FCR) was decreased (p < 0.05) in L1 treatment compared with NC treatment. Overall, the FCR was improved (p < 0.05) in all supplementation treatments compared with NC treatment. The apparently metabolizable nitrogen in L1 treatment was higher (p < 0.05) than that in NC treatment at day 28. The excreta Lactobacillus was increased and E. coli was decreased in PC and L2 treatments compared with NC treatment at day 28 (p < 0.05). The excreta NH₃, H₂S and acetic acid contents were decreased (p < 0.05) in L1 and L2 treatments compared with NC treatment. The relative weight of abdominal fat of broilers fed the PC diet was lowest (p < 0.05) compared with other treatments. In conclusion, this study indicated that dietary supplementation of 0.1% or 0.2% lactulose could improve growth performance, decrease excreta E. coli and excreta NH₃ and H₂S contents.     

An atypical Bacillus anthracis infection in a bull—A potential occupational health hazard

Bacillus anthracis infecting cattle is usually identified based on the typical symptom: sudden death. Bacillus anthracis causing atypical symptoms may remain undiagnosed and represent a potential occupational health hazard for, that is veterinarians and producers, butchers and tanners. In the year 2004, one case of sudden death in a dairy farm in southern Finland was diagnosed as bovine anthrax. Four years later 2008, an atypical case of anthrax was diagnosed in the same holding. The bull was taken to the Production Animal Hospital of the Faculty of Veterinary Medicine, University of Helsinki because of fever, loss of appetite and a symmetrically swollen scrotal sac. Penicillin treatment cured the fever but not the swollen scrotum. Before the intended therapeutic castration, a punctuate consisting of 10 ml fluid collected into a syringe from the scrotal sac was cultivated on blood agar at 37°C. After 24 hr, an almost pure culture of a completely non‐hemolytic Bacillus cereus‐like bacteria was obtained. The strain was identified as B. anthracis using Ba‐specific primers by the Finnish Food Safety Authority (RUOKAVIRASTO). After the diagnosis, the bull was euthanized and destroyed, the personnel were treated with prophylactic antibiotics and the clinic was disinfected. In this particular case, treatment with water, Virkon S and lime seemed to be effective to eliminate endospores and vegetative cells since no relapses of anthrax have occurred in 10 years. This case is the last reported anthrax case in Finland.