Bacteriocin from honeybee beebread Enterococcus avium, active against Listeria monocytogenes

Enterococcus avium isolated from Apis mellifera beebread produces a thermoresistant bacteriocin with a strain-dependent inhibitory effect on Listeria and without effect on gram-negative bacteria. The bacteriocin appeared to be a polypeptide of about 6 kDa. Genetic analyses revealed no extrachromosomal material in E. avium.     

The in vitro antibiofilm activity of selected culinary herbs and medicinal plants against Listeria monocytogenes

Aims:  The antibiofilm activity of extracts obtained from selected herbs, spices, beverages and commercially important medicinal plants was investigated on Listeria monocytogenes .
Methods and Results:  The growth and development of the biofilm was assessed using the crystal violet (CV) assay. The respiratory activity was assessed using the 2, 3-bis [2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) reduction assay. The majority of extracts tested prevented cell adhesion to the polyvinyl chloride (PVC) surface. Seven of the 15 extracts reduced biofilm adhesion of both the clinical and the type strains by at least 50%. In contrast, inhibition of a preformed biofilm was more difficult to achieve, with only three extracts ( Rosmarinus officinalis, Mentha piperita and Melaleuca alternifolia ) inhibiting the growth of both strains by at least 50%.
Conclusions:  Although most extracts were able to reduce initial cell attachment, inhibition of growth in a preformed biofilm was more difficult to achieve.
Significance and Impact of the Study:  The ability to reduce biofilm biomass as shown by several plant extracts warrants further investigation to explore the use of natural products in antibiofilm adhesion.     

Production of antibacterial substances by bifidobacterial isolates from infant stool active against Listeria monocytogenes

AIMS: This study aimed to characterize new isolates of human bifidobacteria, evaluate some of their probiotic potential and to screen these isolates for their effectiveness at inhibiting Listeria monocytogenes in vitro. METHODS AND RESULTS: Thirty-four Bifidobacterium isolates from infant faeces were identified by fructose-6-phosphate phosphoketolase and PCR. Six isolates, coded RBL67, RBL68, RBL69, RBL70, RBL85 and RBL86, showed higher antagonistic activity against L. monocytogenes. Neutralized culture supernatants of these strains did not inhibit L. monocytogenes when tested by agar diffusion method. However, the concentration of supernatant by speed-vac resulted in the formation of an inhibitory effect with supernatants from strains RBL67, RBL68 and RBL70. This effect was shown to be related to heat-stable proteinaceous compound(s) which were resistant to heating at 100 degrees C for 5 min but not to pronase-E, proteinase-K or trypsin. The extraction of the inhibitory compounds by methanol-acetone extraction procedure indicated that four strains (RBL67, RBL68, RBL69 and RBL70) were mostly soluble in acetone. However, strain RBL85 produced inhibitory substances that were soluble in methanol. CONCLUSION: Infant bifidobacterial isolates produce heat-stable proteinaceous compounds active against L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: Production of antibacterial substances by bifidobacteria would improve intestinal bacterial ecology and inhibit intestinal pathogens.     

Highly selective medium for isolation of Listeria monocytogenes from food

A new selective medium (Al-Zoreky-Sandine listeria medium [ASLM]) was formulated to recover Listeria monocytogenes from food specimens; the medium completely inhibited common food microflora. Recognition of Listeria colonies is evident by black discoloration of the medium due to esculin hydrolysis without need for special illuminating equipment. The medium contains acriflavin, ceftazidime, and moxalactam as selective agents. Compared with Listeria Selective Agar, ASLM was equally effective in recovering L. monocytogenes. However, ASLM inhibited micrococci, enterococci, and gram-negative bacteria, especially a strain that mimicked L. monocytogenes on Listeria Selective Agar. The new medium was able to recover heat injured cells with only 15% less count than the nonselective medium.     

Zebrafish bioassay-guided natural product discovery: isolation of angiogenesis inhibitors from East African medicinal plants

Natural products represent a significant reservoir of unexplored chemical diversity for early-stage drug discovery. The identification of lead compounds of natural origin would benefit from therapeutically relevant bioassays capable of facilitating the isolation of bioactive molecules from multi-constituent extracts. Towards this end, we developed an in vivo bioassay-guided isolation approach for natural product discovery that combines bioactivity screening in zebrafish embryos with rapid fractionation by analytical thin-layer chromatography (TLC) and initial structural elucidation by high-resolution electrospray mass spectrometry (HRESIMS). Bioactivity screening of East African medicinal plant extracts using fli-1:EGFP transgenic zebrafish embryos identified Oxygonum sinuatum and Plectranthus barbatus as inhibiting vascular development. Zebrafish bioassay-guided fractionation identified the active components of these plants as emodin, an inhibitor of the protein kinase CK2, and coleon A lactone, a rare abietane diterpenoid with no previously described bioactivity. Both emodin and coleon A lactone inhibited mammalian endothelial cell proliferation, migration, and tube formation in vitro, as well as angiogenesis in the chick chorioallantoic membrane (CAM) assay. These results suggest that the combination of zebrafish bioassays with analytical chromatography methods is an effective strategy for the rapid identification of bioactive natural products.     

Temporal and spatial variation of Botryosphaeriaceae associated with Acacia karroo in South Africa

The Botryosphaeriaceae are common and diverse members of fungal communities infecting woody plants. They are also increasingly being used as model organisms to understand patterns in the global movement of latent pathogens. The aim of this study was to consider the species richness of the Botryosphaeriaceae associated with the native Acacia (Vachellia) karroo across South Africa, and the variation of species at specific local sites over time. The diversity of these fungi associated with different tissues of this host plant was also considered. These questions were addressed by sampling healthy A. karroo from 23 sites in South Africa and by more intensive hierarchical sampling conducted at 40 sites in one area over 3 yr. In total, 13 species of the Botryosphaeriaceae were identified, including seven that were isolated only from the more intensively sampled area. There was a clear geographical influence, with some species occurring only in some parts of the country. Significant variation in the species richness over time for the intensively sampled area was found and there was no evidence of tissue specificity for this group of fungi in leaves, branches and branchlets. Results of pathogenicity trials showed highly variable lesion sizes for the isolated species in comparison to the control, with Sphaeropsis variabilis, Lasiodiplodia theobromae and Neofusicoccum australe being the most pathogenic. The overall results revealed a rich diversity of Botryosphaeriaceae on this native host, which varied significantly both geographically and on individual trees, even in the absence of obvious disease.     

Selective-enrichment procedure for isolation of Listeria monocytogenes from fecal and biologic specimens

A selective-enrichment procedure (SEP) was developed to isolate Listeria monocytogenes from fecal and biologic specimens. This procedure was compared with direct plating with McBride listeria agar and 2-, 4-, and 8-week cold-enrichment procedures in recovering L. monocytogenes from mouse fecal, liver, and brain specimens. Although the SEP occasionally did not isolate the organism from specimens proved positive by the other procedures, the SEP isolated L. monocytogenes from about two and five times as many specimens as the cold-enrichment and direct-plating procedures, respectively.     

Highly selective medium for isolation of Listeria monocytogenes from food.

A new selective medium (Al-Zoreky-Sandine listeria medium [ASLM]) was formulated to recover Listeria monocytogenes from food specimens; the medium completely inhibited common food microflora. Recognition of Listeria colonies is evident by black discoloration of the medium due to esculin hydrolysis without need for special illuminating equipment. The medium contains acriflavin, ceftazidime, and moxalactam as selective agents. Compared with Listeria Selective Agar, ASLM was equally effective in recovering L. monocytogenes. However, ASLM inhibited micrococci, enterococci, and gram-negative bacteria, especially a strain that mimicked L. monocytogenes on Listeria Selective Agar. The new medium was able to recover heat injured cells with only 15% less count than the nonselective medium.     

Proteomic and microscopic analysis of biofilms formed by Listeria monocytogenes 568

Biofilm formation may be important in the colonization of the food-processing environment by the food-borne pathogen Listeria monocytogenes. Listeria monocytogenes 568 formed adherent multicellular layers on a variety of test surfaces following growth at 37 degrees C with multiple transfers of the test surface into fresh medium. Microscopic examination of these adherent layers suggest that the cells were surrounded by extracellular material. The presence of a carbohydrate containing extracellular polymeric matrix was confirmed by labelling hydrated adherent layers with fluorescein-conjugated concanavalin A, indicating that these adherent layers are biofilms. To gain insight into the physiological state of cells in these biofilms, the proteomes from biofilm- and planktonic-grown cells from the same cultures were compared using 2-dimensional polyacrylamide gel electrophoresis. Nineteen proteins, which exhibited higher levels of expression in biofilm-grown cells, were successfully identified from the 2-D gels using a combination of MALDI-TOF and MS/MS. Proteins that were found to be more highly expressed in biofilm-grown cells were involved in stress response, envelope and protein synthesis, biosynthesis, energy generation, and regulatory functions. In biofilm-grown cells, many proteins in the pH range 4-6 ran as multiple spots arranged horizontally across the 2-D gels.     

Enterocin 226NWC, a bacteriocin produced by Enterococcus faecalis 226, active against Listeria monocytogenes

F. VILLANI, G. SALZANO, E. SORRENTINO, O. PEPE, P. MARINO AND S. COPPOLA. 1993. Enterococcus faecalis 226, isolated from natural whey cultures utilized as starters in the manufacture of mozzarella cheese from water-buffalo milk, produces a bacteriocin designated enterocin 226NWC. The bacteriocin was isolated from culture supernatant fluids of the producer strain and was active against strains of the same species and Listeria monocytogenes, but not against useful lactic acid bacteria. Enterocin 226NWC is a protein with an apparent molecular weight of about 5800; it is relatively heat-stable and has a bactericidal mode of action. Listeria monocytogenes, growing in the presence of the enterocin 226NWC producer strain in broth and in reconstituted skim milk, was inhibited.     

Database on pharmacophore analysis of active principles, from medicinal plants

Plants continue to be a major source of medicines, as they have been throughout human history. In the present days, drug discovery from plants involves a multidisciplinary approach combining ethnobotanical, phytochemical and biological techniques to provide us new chemical compounds (lead molecules) for the development of drugs against various pharmacological targets, including cancer, diabetes and its secondary complications. In view of this need in current drug discovery from medicinal plants, here we describe another web database containing the information of pharmacophore analysis of active principles possessing antidiabetic, antimicrobial, anticancerous and antioxidant properties from medicinal plants. The database provides the botanical, taxonomic classification, biochemical as well as pharmacological properties of medicinal plants. Data on antidiabetic, antimicrobial, anti oxidative, anti tumor and anti inflammatory compounds, and their physicochemical properties, SMILES Notation, Lipinski's properties are included in our database. One of the proposed features in the database is the predicted ADMET values and the interaction of bioactive compounds to the target protein. The database alphabetically lists the compound name and also provides tabs separating for anti microbial, antitumor, antidiabetic, and antioxidative compounds. AVAILABILITY: http://www.hccbif.info /     

Selective-enrichment procedure for isolation of Listeria monocytogenes from fecal and biologic specimens.

A selective-enrichment procedure (SEP) was developed to isolate Listeria monocytogenes from fecal and biologic specimens. This procedure was compared with direct plating with McBride listeria agar and 2-, 4-, and 8-week cold-enrichment procedures in recovering L. monocytogenes from mouse fecal, liver, and brain specimens. Although the SEP occasionally did not isolate the organism from specimens proved positive by the other procedures, the SEP isolated L. monocytogenes from about two and five times as many specimens as the cold-enrichment and direct-plating procedures, respectively.     

The potential of South African plants in the development of new medicinal products

Southern Africa is an important focal point of botanical and cultural diversity but only a few plant species have hitherto become fully commercialised as medicinal products. In recent years there has been an upsurge in research and development activity, resulting in several new products and new crops. In this review, more than 90 of the best-known and most promising indigenous South African plants are listed and subjectively evaluated in the context of their potential for commercialisation as medicinal products for a variety of applications. The history of product development relating to the following species is briefly discussed and the plants and some of their products are illustrated: Agathosma betulina (buchu), Aloe ferox (bitter aloe), Artemisia afra (African wormwood), Aspalathus linearis (rooibos tea), Bulbine frutescens (burn jelly plant); Cyclopia genistoides (honeybush tea), Harpagophytum procumbens (devil's claw), Hoodia gordonii (hoodia, ghaap), Hypoxis hemerocallidea (“African potato”), Lippia javanica (fever tea), Mesembryanthemum tortuosum (= Sceletium tortuosum) (kanna, kougoed), Pelargonium sidoides (“Umckaloabo”), Siphonochilus aethiopicus (African ginger), Sutherlandia frutescens (= Lessertia frutescens) (cancer bush), Warburgia salutaris (pepperbark tree) and Xysmalobium undulatum (“Uzara”). The main factors that are apparently responsible for failure or success will be highlighted, especially the importance of marketing strategy, proof of concept and barriers to market entry.     

Cyclooxygenase inhibitory activity of South African plants used against inflammation

During the years 1994–2001 we screened around 200 plant species for cyclooxygenase inhibiting activity. The plants were mostly selected based on usage in traditional medicine for inflammation and pains. Species from 32 genera exhibited high activity against COX-1. From Eucomis autumnalis two compounds, eucomin with COX-1 selectivity and eucosterol with COX-2 selectivity were isolated, a possible reason why this plant is so popular is traditional medicine. Attempts at isolating the compounds responsible for COX inhibition in Ocotea bullata and Siphonochilus aethiopicus revealed that there wasn’t a single active compound present, instead many compounds contributed to the activity. Plants with multiple active components are probably better suited as phytomedicines rather than as sources for pure compound drugs.