The Screening and Evaluation of Fucus serratus and Fucus vesiculosus Extracts against Current Strains of MRSA Isolated from a Clinical Hospital Setting.

Antimicrobial resistant strains of infection are afflicting clinical settings, driving the search for novel antimicrobial compounds. Naturally sourced bioactives, for instance those from seaweeds, have the potential to ameliorate this issue. As such, solvent extracts from the edible Irish seaweeds Fucus serratus and Fucus vesiculosus were screened for antimicrobial activity against 28 clinically isolated strains of MRSA, including one GISA (glycopeptide intermediate S. aureus) and two mecC gene containing strains. The water extract of F. vesiculosus was the most promising extract went on to be tested for biofilm prevention and disruption activity. The disk diffusion method was used to investigate the inhibition of the bacterial pathogens tested while MIC, MBC and biofilm disruption and prevention analyses were performed spectroscopically and by plate counts, respectively. Solvent extracts were found to have a wide array of antimicrobial activity against the strains tested, with the water extract from Fucus vesiculosus being the most promising. This extract was also found to both prevent and disrupt MRSA biofilms indicating the potential extract as new antimicrobials, and raising the possibility of their possible use in therapeutics.

Inactivation of Salmonella during dry co-digestion of food waste and pig manure.

Extremely high volatile fatty acids (VFAs) and ammonia concentrations can accumulate during dry co-digestion of organic wastes, which may inactivate pathogenic microorganisms. In this study, inactivation of Salmonella during dry co-digestion of pig manure (PM) and food waste (FW), which are both reservoirs of zoonotic pathogens, was examined. The effects of pH, VFAs, ammonia and their interactions were assessed on three inoculated Salmonella serotypes. The results show that dry co-digestion significantly decreased the Salmonella inactivation time from several months (in wet digestion) to as short as 6-7 days. A modified Weibull distribution was proposed to simulate Salmonella reduction and to calculate or predict the minimum inhibitory concentrations (MIC) of VFAs and ammonia. Statistical analysis showed that all the factors (pH, VFA type, VFA/ammonia concentration and Salmonella serotype) significantly impacted Salmonella inactivation (P < 0.01). The inhibitory effect sequence was pH > VFA concentration > VFA type > Salmonella serotype in VFA MIC tests, and ammonia concentration > pH > Salmonella serotype in ammonia MIC tests. The toxicity of VFAs was much greater than that of ammonia, and an antagonistic effect was found between VFAs and ammonia on Salmonella inactivation. Apart from the toxicity of free VFAs and free ammonia, the inhibitory effects of pH alone, ionized VFAs and ammonium were also observed.

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

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 S. aureus 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.

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 ß-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.

Evaluation of the efficacy and safety of a marine-derived Bacillus strain for use as an in-feed probiotic for newly weaned pigs.

Forty 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 + 10(10) 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.

Development of a novel antimicrobial seaweed extract-based hydrogel wound dressing.

The objective of this study was to develop a novel antimicrobial seaweed wound dressing. The seaweed extract was active against nine clinically-relevant wound pathogens. A hydrogel formulation was prepared using polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP), followed by addition of 1% seaweed extract. The antimicrobial properties of the novel dressing were tested using agar diffusion assays, with release-profiles examined using gel leaching and gel transfer assays. The dressing was found to be effective against the same microbial strains as the seaweed extract, with similar efficacy to the commonly used silver-based dressing, Acticoat(®). Antimicrobial release-profile assays revealed that the dressing was effective in inhibiting 70-90% of the bacterial population within the first 30 min, followed by a long, sustained released up to 97 h, without leaving a residue following five subsequent transfers of the dressing. Antimicrobial activity was stable for up to 6 months of storage at 4 °C, but activity was reduced slightly after 15 weeks. Following autoclave sterilization, the dressing displayed a slower release profile compared to a non-autoclaved counterpart. Hence, the seaweed dressing may have commercial applications, potentially competing with silver-based dressings at a lower cost per-application. This is the first report of development of a seaweed-based antimicrobial dressing.

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.

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.

Antioxidative, tyrosinase inhibiting and antibacterial activities of leaf extracts from medicinal ferns.

Leaf extracts of five medicinal ferns, Acrostichum aureum L. (Pteridaceae), Asplenium nidus L. (Aspleniaceae), Blechnum orientale L. (Blechnaceae), Cibotium barometz (L.) J. Sm. (Cyatheaceae) and Dicranopteris linearis (Burm.) underwood var. linearis (Gleicheniaceae), were investigated for their total phenolic content (TPC), and antioxidative, tyrosinase inhibiting and antibacterial activities. The antioxidative activity was measured by assays for radical scavenging against 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric ion reducing power (FRP), beta-carotene bleaching (BCB) and ferrous ion chelating (FIC). The results revealed B. orientale to possess the highest amount of total polyphenols and strongest potential as a natural antioxidative, tyrosinase inhibiting and antibacterial agent as demonstrated by its strong activities in all related bioassays. The other ferns with antioxidative potential were C. barometz and D. linearis. Except for A. aureum, all ferns showed antibacterial activity which may justify their usage in traditional medicines.