Biosurfactant production by halophilic yeasts isolated from extreme environments in Botswana.

Nine morphologically distinct halophilic yeasts were isolated from Makgadikgadi and Sua pans, as pristine and extreme environments in Botswana. Screening for biosurfactant production showed that Rhodotorula mucilaginosa SP6 and Debaryomyces hansenii MK9 exhibited the highest biosurfactant activity using Xanthocercis zambesiaca seed powder as a novel and alternative inexpensive carbon substrate. Chemical characterization of the purified biosurfactants by Fourier Transform Infra-Red spectroscopy suggested that the biosurfactant from R. mucilaginosa SP6 was a rhamnolipid-type whereas the biosurfactant from D. hansenii MK9 was a sophorolipid-type. The two biosurfactants exhibited antimicrobial activities against eight pathogenic bacteria and fungal strains (Proteus vulgaris, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Micrococcus luteus, Cryptococcus neoformans, Candida albicans and Aspergilus niger). The sophorolopid-type biosurfactant was found to be the most potent among the antimicrobial drug resistant strains tested. The findings open up prospects for the development of environmentally friendly antimicrobial drugs that use an inexpensive source of carbon to reduce the costs associated with the production of biosurfactants.

On the use of n-octyl gallate and salicylhydroxamic acid to study the alternative oxidase role.

The alternative oxidase (AOX) catalyzes the transfer of electrons from ubiquinol to oxygen without the translocation of protons across the inner mitochondrial membrane. This enzyme has been proposed to participate in the regulation of cell growth, sporulation, yeast-mycelium transition, resistance to reactive oxygen species, infection, and production of secondary metabolites. Two approaches have been used to evaluate AOX function: incubation of cells for long periods of time with AOX inhibitors or deletion of AOX gene. However, AOX inhibitors might have different targets. To test non-specific effects of n-octyl gallate (nOg) and salicylhydroxamic acid (SHAM) on fungal physiology we measured the growth and respiratory capacity of two fungal strains lacking (Ustilago maydis-Δaox and Saccharomyces cerevisiae) and three species containing the AOX gene (U. maydis WT, Debaryomyces hansenii, and Aspergillus nidulans). For U. maydis, a strong inhibition of growth and respiratory capacity by SHAM was observed, regardless of the presence of AOX. Similarly, A. nidulans mycelial growth was inhibited by low concentrations of nOg independently of AOX expression. In contrast, these inhibitors had no effect or had a minor effect on S. cerevisiae and D. hansenii growth. These results show that nOg and SHAM have AOX independent effects which vary in different microorganisms, indicating that studies based on long-term incubation of cells with these inhibitors should be considered as inconclusive.

Lactobacillus delbrueckii subsp. bulgaricus F17 and Leuconostoc lactis H52 supernatants delay the decay of strawberry fruits: a microbiome perspective.

Strawberries are vulnerable to physical injuries and microbial invasion. To explore if beneficial lactic acid bacteria can improve the shelf life and edible quality of postharvest strawberry fruits, the effects of Lactobacillus delbrueckii subsp. bulgaricus (ital.) F17 (F17) and Leuconostoc lactis (ital.) H52 (H52) inoculation on the strawberry microbial community structure and saleable characteristics were examined by bacterial 16S rRNA and fungal ITS sequencing techniques. Lactobacillus (ital.) F17 and Leuconostoc lactis (ital.) H52 isolated from the traditional fermented yak milk in the Qinghai-Tibetan Plateau were used as the potential probiotic inocula. Samples from treated strawberries stored at 25 °C for 0, 12, 24, 48, and 72 hours were analyzed for their pH, weight loss percentage, decay percentage, total soluble solid content (SSC) and microbial counts, and for microbiome community diversity and canonical correspondence analysis. The results showed that F17 and H52 did not only significantly reduce the weight loss and decay percentage of strawberry fruits, but also delayed the decrease of the total SSC and pH (P < 0.05). In addition, F17 and H52 significantly inhibited the growth and colonization of aerobic mesophilic bacteria, yeast, mold and coliform bacteria. In particular, by comparing the microbiota composition of the samples, F17 significantly inhibited Pantoea, Mycospherella, unclassified_Pleosporales, Aureobasidium and Phoma at the genus level, whereas H52 inhibited Bacillus, Streptophyta, Mycospherella, Aureobasidium and Phoma. Moreover, analysis of alpha and beta diversity revealed that F17 and H52 had a significantly greater inhibitory effect on bacterial species compared to fungi. The results of canonical correspondence analysis revealed that the total SSC and pH were positively correlated with bacteria, whereas the decay percentage, weight loss percentage and total SSC were positively associated with fungi. Additionally, Podosphaera, Hanseniaspora, Botrytis and unclassified_Pleosporales were positively correlated with strawberry fruit decay and weight loss percentage. As a general result, Lactobacillus F17 and Leuconostoc lactis H52 have the potential to promote biological preservation, which is economically important to reduce the loss due to strawberry spoilage.

Bactericidal and Fungicidal Activity in the Gas Phase of Sodium Dichloroisocyanurate (NaDCC).

Sodium dichloroisocyanurate (NaDCC) is usually employed as a disinfectant for the treatment of water, environmental surfaces and medical equipment principally for its effectiveness as a microbicide agent. In this study, we explore the possibility of a new use for NaDCC by investigating the microbicidal activity of chlorine, which derives from the hydrolysis of NaDCC mediated by air humidity, and by testing its effect on the neutralization of microbes present in domestic waste. NaDCC was inserted in a plastic garbage can where LB agar plates, with different dilutions of a known title of four different microorganisms (Escherichia coli, Staphylococcus aureus, Debaryomyces hansenii and Aspergillus brasiliensis), were weakly inserted. The molecular chlorine (Cl2) levels present in the garbage can were quantified using an iodometric titration. The gas emitted in the garbage can presented a strong microbicide effect, inhibiting the proliferation of all four microorganisms and for four consecutive weeks, thus showing that NaDCC hydrolysis, mediated by air humidity, is able to ensure the decontamination of restricted environments, avoiding the proliferation of both Gram-positive and Gram-negative bacteria as well as fungi.

Antifungal properties of durancins isolated from Enterococcus durans A5-11 and of its synthetic fragments.

The aim of this work was to study the antifungal properties of durancins isolated from Enterococcus durans A5-11 and of their chemically synthesized fragments. Enterococcus durans A5-11 is a lactic acid bacteria strain isolated from traditional Mongolian airag cheese. This strain inhibits the growth of several fungi including Fusarium culmorum, Penicillium roqueforti and Debaryomyces hansenii. It produces two bacteriocins: durancin A5-11a and durancin A5-11b, which have similar antimicrobial properties. The whole durancins A5-11a and A5-11b, as well as their N- and C-terminal fragments were synthesized, and their antifungal properties were studied. C-terminal fragments of both durancins showed stronger antifungal activities than other tested peptides. Treatment of D. hansenii LMSA2.11.003 strain with 2 mmol l(-1) of the synthetic peptides led to the loss of the membrane integrity and to several changes in the ultra-structure of the yeast cells. Chemically synthesized durancins and their synthetic fragments showed different antimicrobial properties from each other. N-terminal peptides show activities against both bacterial and fungal strains tested. C-terminal peptides have specific activities against tested fungal strain and do not show antibacterial activity. However, the C-terminal fragment enhances the activity of the N-terminal fragment in the whole bacteriocins against bacteria.

Antimicrobial activities of neo- and 1-epineo-isoshinanolones from Plumbago zeylanica roots.

CONTEXT: The roots of Plumbago zeylanica Linn. (Plumbaginaceae) are reputed to have a wide spectrum of therapeutic properties in the Ayurvedic system of medicine. They are useful in curing many ailments such as skin diseases, diarrhea, plague and leprosy. OBJECTIVE: The study was aimed at isolating, separating and evaluating the antimicrobial properties of compounds such as neoisoshinanolone and 1-epineo-isoshinanolone from the roots of P. zeylanica. MATERIALS AND METHODS: The crude petroleum ether extract of roots of P. zeylanica was subjected to repeated chromatographic techniques to separate compounds 2 and 3 along with plumbagin. Structure elucidation was carried out using nuclear magnetic resonance (NMR), infra red (IR) and mass spectroscopy. The serial dilution method was used to test antimicrobial activities and their minimum inhibitory concentration (MIC) expressed in microg/mL. RESULTS: 1-Epineo-isoshinanolone is more active with a MIC of 12.5-25 microg/mL whereas neoisoshinanolone has recorded a MIC of 50-100 microg/mL. The activities are compared with plumbagin (0.78-3.13 microg/mL) and standards streptomycin for bacteria and nystatin for fungi. DISCUSSION: Earlier researchers have established the presence of plumbagin in the roots of P. zeylanica and its antimicrobial activities. The structure elucidation of two more biologically active biogenetic precursors along with their activities in the root extracts has been established for the first time in the present study. CONCLUSION: The root extract of P. zeylanica possesses good antimicrobial activity, which suggests its therapeutic use in the Ayurvedic system of medicine to cure skin diseases.

Effect of mixed antimicrobial agents and flavors in active packaging films.

Active packaging is an emerging food technology to improve the quality and safety of food products. Many works have been developed to study the antimicrobial activity of essential oils. Essential oils have been traditionally used as flavorings in food, so they have an important odor impact but they have as well antimicrobial properties that could be used to protect the food. Recent developments in antimicrobial active packaging showed the efficiency of essential oils versus bread and bakery products among other applications. However, one of the main problems to face is the odor and taste they could provide to the packaged food. Using some aromas to mask the odor could be a good approach. That is why the main objective of this paper is to develop an antimicrobial packaging material based on the combination of the most active compounds of essential oils (hydrocinnamaldehyde, oregano essential oil, cinnamaldehyde, thymol, and carvacrol) together with some aromas commonly used in the food industry. A study of the concentration required to get the antimicrobial properties, the organoleptic compatibility with typical aroma present in many food systems (vanilla, banana, and strawberry), and the right combination of both systems has been carried out. Antimicrobial tests of both the mentioned aromas, the main components of some essential oils, and the combination of both groups were carried out against bacteria (Enterococcus faecalis, Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Salmonella choleraesuis, Yersinia enterocolitica, Escherichia coli), yeasts (Candida albicans, Debaryomyces hansenii, Zygosaccharomyces rouxii), and molds (Botrytis cinerae, Aspergillus flavus, Penicillium roqueforti, Eurotium repens, Penicillium islandicum, Penicillium commune, Penicillium nalgiovensis). The sensory properties of the combinations were evaluated with a triangular test and classification was by an order test; the odor threshold of the aroma compounds was also studied. The results reveal that none of the aromas had antimicrobial properties. The most antimicrobial compounds are thymol, carvacrol, and cinnamaldehyde, but none of them could be combined with banana aroma, whereas only thymol with strawberry aroma gave the right combined organoleptic profile. All of the antimicrobials under study could be combined with vanilla aroma, providing both antimicrobial property and the odor expected.

Effects of calcium cyanamide on soil microbial communities and Fusarium oxysporum f. sp. cucumberinum.

Calcium cyanamide (CaCN(2)) has been one of the potential candidates as soil disinfectant since the restriction of methyl bromide in soil fumigation due to its ecological risk. However, little information is available on effects of CaCN(2) on soil microbial community. In this study, the soil microbial communities and the fate of pathogen Fusarium oxysporum (Schlechtend, Fr) f. sp. cucumberinum (Owen) Snyder and Hansen (F.O. f. sp. cucumberinum) in response to CaCN(2) treatment was evaluated. F.O. f. sp. cucumberinum population in soil treated with CaCN(2) at rates of 80 and 200 gm(-2) was suppressed by 88.7 and 92.2% after 15 d of CaCN(2) application. Bacterial, fungal, and actinomycete populations were also greatly decreased after 3 d of CaCN(2) application, but they recovered to the control level by 15 d. The variation in functional diversity of soil microbes characterized by principal component analysis, diversity and evenness indices based on Biolog data followed a similar trend. Meanwhile, the band number from the DGGE of soil 16S rDNA fragments increased from 9 for the non-CaCN(2)-treated soil to 10 or 12 after different rates of CaCN(2) application at 15 d, indicating the increase of abundant rDNA types in the community. The results suggest that CaCN(2) application had only a short-term and transitory impact on the indigenous soil microbial community in contrast to the long-term suppression of the F.O. f. sp. cucumberinum population. It is feasible to reduce Fusarium wilt without significant impact on microbial community by application of CaCN(2) at reasonable doses.

Synthesis, Raman, FT-IR, NMR spectroscopic data and antimicrobial activity of mixed aza-oxo-thia macrocyclic compounds.

Mixed aza-oxo-thia macrocyclic ligands 1,3,5,11,13,15-hexaaza-6,10,16,20-tetraoxo-8,18-dithia-2,3,4:12,13,14-dipyridine cyclocosane (L(1)); 1,3,5,12,14,16-hexeaza-6,11,17,22-tetraoxo-8,9,19,20-tetrathia-2,3,4:13,14,15-dipyridine cyclodocosane (L(2)); 1,3,5,13,15,17-hexaaza-6,12,18,24-tetraoxo-9,21-dithia-2,3,4:14,15,16-dipyridine cyclotetracosane (L(3)) and 1,3,5,14,16,18-hexaaza-6,13,19,26-tetraoxo-9,10,22,23-tetrathia-2,3,4:15,16,17-dipyridine cyclohexacosane (L(4)) were synthesised. The structural features of the ligands have been studied by elemental analyses, Raman, IR, (1)H and (13)C NMR spectroscopy. The antimicrobial activities of the ligands were evaluated using disk diffusion method in dimethyl sulfoxide (DMSO) as well as the minimal inhibitory concentration (MIC) dilution method, against nine bacteria. The obtained results from disk diffusion method were assessed in side-by-side comparison with those of penicillin G, ampicillin, cefotaxime, vancomycin, ofloxacin, and tetracycline well known antibacterial agents. The results from dilution procedure were compared with gentamycin as antibacterial and nystatin as antifungal. The antifungal activities are reported on five yeast cultures namely Candida albicans, Kluyveromyces fragilis, Rhodotorula rubra, Debaryomyces hansenii, and Hanseniaspora guilliermondii, and the results are referenced with nystatin, Ketoconazole, and clotrimazole, commercial antifungal agents. In most cases, the compounds show broad-spectrum (Gram(+) and Gram(-) bacteria) activities that were more active or equipotent to the antibiotic and antifungal agents in the comparison tests.

Inhibitory activity against plant pathogenic fungi of extracts from Myoporum bontioides A. Gray and indentification of active ingredients.

BACKGROUND: In order to understand the bioactivity of Myoporum bontioides A. Gray against plant pathogens and determine its active ingredients, the inhibitory activities of methanol extracts from M. bontioides against Fusarium oxysporum f. sp. niveum (E. F. Smith) Snyder & Hansen, Pestalotia mangiferae P. Henn., Thielaviopsis paradoxa (De Seynes) v. Hohnel, Colletotrichum musae (Berk. & M. A. Curtis) v. Arx, Alternaria alternata (Fr.) Keissler, Mycosphaerella sentina (Fr.) Schroter and Sphaceloma fawcettii Jenk. were evaluated using a growth rate method, and the active ingredient was isolated by activity-directed isolation and identified by determination and analysis of IR, (1)H NMR, (13)C NMR and mass spectra and correlative physical constants. RESULTS: The results showed that the extracts from stems and leaves of M. bontioides exhibited inhibitory activity against the seven fungi, with > 58% inhibition at 10 g L(-1) after 72 h. The active compound was isolated and identified as (-)-epingaione, and showed inhibitory activity against the above seven fungi. The inhibitory activity against P. mangiferae was the highest, with an EC(50) value of 77 mg L(-1). The EC(50) values against the other six fungi were 147-245 mg L(-1). (-)-Epingaione also inhibited spore germination of F. oxysporum f. sp. niveum, T. paradoxa and S. fawcettii. CONCLUSION: (-)-Epingaione demonstrated broad-spectrum inhibitory activity against plant pathogenic fungi and is promising for exploitation as a fungicide.

Antibiotic activity of the pyrenocines.

Pyrenocine A, a phytotoxin produced by Pyrenochaeta terrestris (Hansen) Gorenz, Walker and Larson, possesses general antibiotic activity against plants, fungi, and bacteria. Effective doses for 50% inhibition (ED50s) are 4 micrograms/mL for onion seedling elongation; 14, 20, 20, and 25 micrograms/mL for the germination of asexual spores of Fusarium oxysporum f. sp. cepae, Fusarium solani f. sp. pisi, Mucor hiemalis, and Rhizopus stolonifer, respectively. Pyrenocine A also inhibits the linear mycelial growth of both P. terrestris and F. oxysporum with ED50s calculated as 77 and 54 micrograms/mL, respectively. Gram-positive bacteria are more susceptible to pyrenocine A than Gram-negative bacteria. ED50s are estimated as 30, 45, and 200 micrograms/mL for the inhibition of growth of Bacillus subtilis, Staphylococcus aureus, and Escherichia coli, respectively, with Pseudomonas aeruginosa resistant to those concentrations tested. Pyrenocine A acts primarily as a biostatic rather than a biocidal agent with all organisms tested showing some degree of recovery when released from pyrenocine A. Pyrenocines B and C show little antibiotic activity in the bioassays performed.