Evaluation of the Efficacy and Safety of a Marine-Derived Bacillus Strain for Use as an In-Feed Probiotic for Newly Weaned Pigs

peer-reviewedForty eight individual pigs (8.7±0.26 kg) weaned at 28±1 d of age were used in a 22-d study to evaluate the effect of oral administration of a Bacillus pumilus spore suspension on growth performance and health indicators. Treatments (n = 16) were: (1) non-medicated diet; (2) medicated diet with apramycin (200 mg/kg) and pharmacological levels of zinc oxide (2,500 mg zinc/kg) and (3) B. pumilus diet (non-medicated diet + 1010 spores/day B. pumilus). Final body weight and average daily gain tended to be lower (P = 0.07) and feed conversion ratio was worsened (P<0.05) for the medicated treatment compared to the B. pumilus treatment. Ileal E. coli counts were lower for the B. pumilus and medicated treatments compared to the non-medicated treatment (P<0.05), perhaps as a result of increased ileal propionic acid concentrations (P<0.001). However, the medicated treatment reduced fecal (P<0.001) and cecal (P<0.05) Lactobacillus counts and tended to reduce the total cecal short chain fatty acid (SCFA) concentration (P = 0.10). Liver weights were lighter and concentrations of liver enzymes higher (P<0.05) in pigs on the medicated treatment compared to those on the non-medicated or B. pumilus treatments. Pigs on the B. pumilus treatment had lower overall lymphocyte and higher granulocyte percentages (P<0.001) and higher numbers of jejunal goblet cells (P<0.01) than pigs on either of the other two treatments or the non-medicated treatment, respectively. However, histopathological examination of the small intestine, kidneys and liver revealed no abnormalities. Overall, the B. pumilus treatment decreased ileal E. coli counts in a manner similar to the medicated treatment but without the adverse effects on growth performance, Lactobacillus counts, cecal SCFA concentration and possible liver toxicity experienced with the medicated treatment.The study was funded by the Higher Education Authority/Institutes of Technology Ireland Technological Sector Research Strand III Programme

Choosing Short: An Explanation of the Similarities and Dissimilarities in the Distribution Patterns of Binding and Covaluation

Covaluation is the generalization of coreference introduced by Tanya Reinhart. Covaluation distributes in patterns that are very similar yet not entirely identical to those of binding. On a widespread view, covaluation and binding distribute similarly because binding is defined in terms of covaluation. Yet on Reinhart's view, binding and covaluation are not related that way: binding pertains to syntax, covaluation does not. Naturally, the widespread view can easily explain the similarities between binding and covaluation, whereas Reinhart can easily explain the dissimilarities. Reciprocally, the widespread view finds it harder to explain the dissimilarities, whereas Reinhart finds it harder to explain the similarities. Reinhart and others have proposed more than one explanation of the similarities, but as I argue, these explanations do not work. Hence although I adopt Reinhart's view, I propose a new explanation of the similarities and dissimilarities between binding and covaluation: While Reinhart has invoked semantic structure only to explain dissimilarities, I do so to explain both similarities and dissimilarities at once. Finally, I examine in light of this approach the topics of language acquisition, only-constructions, the identity predicate, the Partee/Bach/Higginbotham problem, the Dahl puzzle and its recent versions by Roelofsen

Standardised herbal extract of chlorogenic acid from leaves of Etlingera elatior (Zingiberaceae)

Background: Chlorogenic acid (CGA) or 5-caffeoylquinic acid, was found to be the dominant phenolic compound in leaves of Etlingera elatior (Zingiberaceae). The CGA content of E. elatior leaves was significantly higher than flowers of Lonicera japonica (honeysuckle), the commercial source. In this study, a protocol to produce a standardised herbal CGA extract from leaves of E. elatior using column chromatography was developed. Materials and Methods: Freeze-dried leaves of E. elatior were extracted with 30% ethanol, and sequentially fractionated using Diaion HP-20 and Sephadex LH-20. Results: The CGA fractions, which yielded extracts of 10% and 40% w/w purity, possessed antioxidant, tyrosinase inhibition, and antibacterial properties. The entire fractionation process took only 6.5 hours, using gravity flow. From 50 g of leaves, the final yield of CGA extract was 0.2 g (0.4%). The CGA content of the standardised herbal extract from leaves of E. elatior (40%) is 1.6 times that of commercial extracts from honeysuckle flowers (25%). Conclusion: With high CGA content, the standardised herbal extract has a great potential to be developed into functional food and other health products. Leaves of E. elatior, which currently have no economic value, could serve as an alternative source of CGA. Leaves are large, available in abundance, and harvesting is non-destructive to the plants

Assessment of the antioxidant and antibacterial activities of different olive processing wastewaters

<div><p>Olive processing wastewaters (OPW), namely olive mill wastewater (OMW) and table-olive wastewaters (TOW) were evaluated for their antibacterial activity against five Gram-positive and two Gram-negative bacteria using the standard disc diffusion and thin layer chromatography (TLC)-bioautography assays. Disc diffusion screening and bioautography of OMW were compared to the phenolic extracts of table-olive brines. Positive activity against <i>S</i>. <i>aureus</i> was demonstrated. The optimization of chromatographic separation revealed that hexane/acetone in the ratio of 4:6 was the most effective for phenolic compounds separation. A HPLC-MS analysis was performed showing that only two compounds, hydroxytyrosol and tyrosol, were the predominant phenolic compounds in all OPW. The phenolic extract of OMW generated by a semi-modern process showed the highest free radical-scavenging activity (DPPH assay) compared to the other phenolic extracts. It is apparent from the present study that OPW are a rich source of antioxidants suitable for use in food, cosmetic or pharmaceutical applications.</p></div

Effect of concentration of OMW and TOW phenolic extracts on their antibacterial activity against <i>S</i>. <i>aureus</i> using the disc diffusion assay.

<p>Effect of concentration of OMW and TOW phenolic extracts on their antibacterial activity against <i>S</i>. <i>aureus</i> using the disc diffusion assay.</p

Physicochemical characterization of olive mill wastewater and table-olive brine samples.

<p>Physicochemical characterization of olive mill wastewater and table-olive brine samples.</p

HPLC chromatograms of OMW phenolic extracts.

<p>1. Hydroxytyrosol, 2. 3,4-dihydroxyphenylacetic acid, 3. Tyrosol. 4. Protocatechuic acid derivative, 5. Caffeic acid. 6. <i>p</i>-coumaric acid. 7. Ferulic acid derivative, 8. Ferulic acid, 9. Luteolin derivative, 10. Oleuropein. Peaks 1, 3, 6, 8 and 10 were identified by use of standards. The remaining peaks were tentatively identified by comparison with literature data. (OMW1: olive mill wastewater from semi-modern process, OMW2: olive mill wastewater from modern process, GTOW: green table-olive brine, PTOW: purple table-olive brine and BTOW: black table-olive brine).</p

Total phenolic content, phenolic constituents and antioxidant activity (IC₅₀) of different olive processing wastewaters samples.

<p>The values in the brackets are the proportions to the total phenolic content, TPC.</p

In Vitro Assessment of Marine Bacillus for Use as Livestock Probiotics

Six antimicrobial-producing seaweed-derived Bacillus strains were evaluated in vitro as animal probiotics, in comparison to two Bacillus from an EU-authorized animal probiotic product. Antimicrobial activity was demonstrated on solid media against porcine Salmonella and E. coli. The marine isolates were most active against the latter, had better activity than the commercial probiotics and Bacillus pumilus WIT 588 also reduced E. coli counts in broth. All of the marine Bacillus tolerated physiological concentrations of bile, with some as tolerant as one of the probiotics. Spore counts for all isolates remained almost constant during incubation in simulated gastric and ileum juices. All of the marine Bacillus grew anaerobically and the spores of all except one isolate germinated under anaerobic conditions. All were sensitive to a panel of antibiotics and none harbored Bacillus enterotoxin genes but all, except B. pumilus WIT 588, showed some degree of β-hemolysis. However, trypan blue dye exclusion and xCELLigence assays demonstrated a lack of toxicity in comparison to two pathogens; in fact, the commercial probiotics appeared more cytotoxic than the majority of the marine Bacillus. Overall, some of the marine-derived Bacillus, in particular B. pumilus WIT 588, demonstrate potential for use as livestock probiotics

Assessment of the Bacteriocinogenic Potential of Marine Bacteria Reveals Lichenicidin Production by Seaweed-Derived Bacillus spp.

The objectives of this study were (1) to assess the bacteriocinogenic potential of bacteria derived mainly from seaweed, but also sand and seawater, (2) to identify at least some of the bacteriocins produced, if any and (3) to determine if they are unique to the marine environment and/or novel. Fifteen Bacillus licheniformis or pumilus isolates with antimicrobial activity against at least one of the indicator bacteria used were recovered. Some, at least, of the antimicrobials produced were bacteriocins, as they were proteinaceous and the producers displayed immunity. Screening with PCR primers for known Bacillus bacteriocins revealed that three seaweed-derived Bacillus licheniformis harbored the bli04127 gene which encodes one of the peptides of the two-peptide lantibiotic lichenicidin. Production of both lichenicidin peptides was then confirmed by mass spectrometry. This is the first definitive proof of bacteriocin production by seaweed-derived bacteria. The authors acknowledge that the bacteriocin produced has previously been discovered and is not unique to the marine environment. However, the other marine isolates likely produce novel bacteriocins, as none harboured genes for known Bacillus bacteriocins