Optimization of the biological component of a bioelectrochemical cell

The efficiency of electron transduction by Shewanella oneidensis MR-1 was investigated both in batch culture and in a dual-chambered electrochemical cell. Aerobically grown bacteria were inoculated into an insoluble FeOOH suspension in an anaerobic environment. As the bacteria reduced Fe(III) to Fe(II) there was a visible color change from red to bluish black; this simple color change assay proved to be a robust method for determining the electrochemical activity of S. oneidensis MR-1. In an effort to improve electricity production by S. oneidensis MR-1, the performance of the electrochemical cell with lactate as a fuel was first optimized with respect to both poised potential and fuel concentration. Ultimately, electricity production proved to be proportional to both the poised potential and to fuel concentration. In particular, higher poised potentials increased charge production. Finally, we attempted to optimize the bacteria themselves for the efficient transduction of reduced carbon sources into electricity. The batch culture underwent a series of serial dilutions; after the 4th dilution the microbe population exhibited a 30% increase in charge production. We are further exploring whether this increase was due to metabolic adaptations or to genetic mutations, and examining additional ways to evolve electrogenic organisms.     

Resistance of Pseudomonas aeruginosa to liquid disinfectants on contaminated surfaces before formation of biofilms

A comparison was made of the effectiveness of popular disinfectants (Cavicide, Cidexplus, Clorox, Exspor, Lysol, Renalin, and Wavicide) under conditions prescribed for disinfection in the respective product labels on Pseudomonas aeruginosa either in suspension or deposited onto surfaces of metallic or polymeric plastic devices. The testing also included 7 nonformulated germicidal agents (glutaraldehyde, formaldehyde, peracetic acid, hydrogen peroxide, sodium hypochlorite, phenol, and cupric ascorbate) commonly used in disinfection and decontamination. Results showed that P. aeruginosa is on average 300-fold more resistant when present on contaminated surfaces than in suspension. This increase in resistance agrees with results reported in studies of biofilms, but unexpectedly, it precedes biofilm formation. The surface to which bacteria are attached can influence the effectiveness of disinfectants. Viable bacteria attached to devices may require dislodging through more than a one-step method for detection. The data, obtained with a sensitive and quantitative test, suggest that disinfectants are less effective on contaminated surfaces than generally acknowledged.     

Microbiological methods for testing disinfectant efficiency on Pseudomonas biofilm

Biofilms of the Gram-negative bacteria Pseudomonas aeruginosa and Pseudomonas fragi were grown on stainless steel surfaces (AISI 304, 2B) for 4 days in slime broth. These biofilms were treated with four commercial disinfectants. The disinfectants were alcohol-based, tenside-based, peroxide-based and chlorine-based products, covering most disinfectant types used in the food industry. The effects of the disinfectants on the bacterial cells were first investigated in suspension using the permeabilisation test, which is based on fluorescence assessment of hydrophobic 1-N-phenyl-naphtylamine (NPN). The surfaces covered with disinfectant-treated biofilms were investigated using conventional cultivation, impedimetry and epifluorescence microscopy in combination with image analysis of preparations stained with the DNA-stain acridine orange and with the metabolic indicator system CTC-DAPI. The results showed that the tenside-based and peroxide-based disinfectants permeabilised the cells in suspension. The overall biofilm results showed that of the agents tested, the peroxide-based and chlorine-based disinfectants acted most effectively on cells in biofilms.     

Evaluating the Antifungal Potential of Botanical Compounds to Control Botryotinia fuckeliana and Rhizoctonia solani

The European Union is promoting regulatory changes to ban fungicides because of the impact their use has on the ecosystem and the adverse effects they can pose for humans. An ecofriendly alternative to these chemicals to fight against fungal species with low toxicity is essential oils and their compounds extracted from aromatic plants. The purpose of this study was to evaluate the in vitro antifungal capacity of the botanical compounds eugenol, carvacrol, thymol, and cinnamaldehyde, and the synergy or antagonism of their mixtures, against Botryotinia fuckeliana and Rhizoctonia solani. Different bioassays were performed at doses of 300, 200, 150, and 100 µg/mL using pure commercial compounds and their combination in potato dextrose agar culture medium. Growth rate and the mycelium growth inhibition parameters were calculated. Phenolic compounds and their combination inhibited the development of species at the different concentrations, with fungicidal or fungistatic activity shown under almost all the tested conditions. When comparing the growth rates of the species in the control plates and treatments, the statistical analysis showed that there were statistically significant differences. The mixture of compounds improved fungicidal activity against the studied species and at a lower concentration of monoterpenes.     

Antibacterial effect of silver nanoparticles prepared in bipolymers at moderate temperature

The purpose of this study was to investigate the antibacterial effect of silver nanoparticles in chitosan–poly(ethylene glycol) suspension. The silver nanoparticles (AgNPs) were prepared by use of an environmentally benign method from chitosan (Cts) and poly(ethylene glycol) (PEG) at moderate temperature and with stirring for different times. Silver nitrate (AgNO₃) was used as the metal precursor and Cts and PEG were used as solid support and polymeric stabilizer, respectively. The antibacterial activity of silver–chitosan–poly(ethylene glycol) nanocomposites (Ag–Cts–PEG NCs) against Staphylococcus aureus, Micrococcus luteum, Pseudomonas aeruginosa, and Escherichia coli was tested by use of the Mueller–Hinton agar disk-diffusion method. Formation of AgNPs was determined by UV–visible spectroscopy; surface plasmon absorption maxima were observed at 415–430 nm in the UV–visible spectrum. The peaks in the XRD pattern confirmed that the AgNPs had a face-centered cubic structure; peaks of contaminated crystalline phases were not observed. Transmission electron microscopy (TEM) revealed that the AgNPs synthesized were spherical. The optimum stirring time for synthesis of the smallest particle size (mean diameter 5.50 nm) was 12 h. The AgNPs in Cts–PEG were effective against all the bacteria tested. Higher antibacterial activity was observed for AgNPs with smaller size. These results suggest that AgNPs can be used as an effective inhibitor of bacteria and can be used in medical applications. These results also suggest that AgNPs were successfully synthesized in Cts–PEG suspension at moderate temperature with different stirring times.     

Antifungal Activity of Diketopiperazines and Stilbenes Against Plant Pathogenic Fungi In Vitro

The present study aimed to investigate antifungal activity of a stilbene and diketopiperazine compounds against plant pathogenic fungi, including Phytophthora capsici, P. colocasiae, Botrytis cinerea and Colletotrichum gloeosporioides. Minimal inhibition concentrations (MIC) and minimal fungicidal concentrations (MFC) of stilbenes and diketopiperazines for each fungus were determined using microplate method. Best activity was recorded by stilbenes against P. capsici and P. colocasiae. All four test compounds were effective in inhibiting different stages of the life cycle of test fungi. Stilbenes were more effective than diketopiperazines in inhibiting mycelial growth and inhibiting different stages of the life cycle of P. capsici and P. colocasiae. Rupture of released zoospores induced by stilbenes was reduced by addition of 100 mM glucose. The effects of stilbenes on mycelial growth and zoospore release, but not zoospore rupture, were reduced largely when pH value was above 7. In addition, stilbenes were investigated for its antifungal stability against Phytophthora sp. The results showed that stilbenes maintained strong fungistatic activity over a wide pH range (pH 4–9) and temperature range (70–120 °C). The compound stilbenes exhibited strong and stable broad-spectrum antifungal activity, and had a significant fungicidal effect on fungal cells. Results from prebiocontrol evaluations performed to date are probably useful in the search for alternative approaches to controlling serious plant pathogens.     

Gas/gas and gas/wall average energy transfer from very low-pressure pyrolysis

It is shown that data obtained using very low-pressure pyrolysis (VLPP) on the pressure and temperature dependence of unimolecular rate coefficients of reactants with several reaction channels yield average energies transferred in gas/gas and gas/wall collisions (the wall being seasoned quartz at 800–1200 K). The downward average energy transferred, «ΔEå, for chlorocyclobutane/ethylene collisions is found to be 1600 cm^?1 at 970 K; «ΔEå for chlorocyclobutane/wall collisions varies from 5000 cm^?1 (wall efficiency β_w = 0.8) at 930 K to 3500 cm^?1 (β_w = 0.4) at 1150 K; similar values are found from published data on cycloheptatriene and cyclopropane-d₂. This indicates that the assumption of unit wall efficiency usually used in fitting VLPP experiments to RRKM theory needs revision.     

Total Aseptization of Boar Semen,to Increase the Biosecurity of Reproduction in Swine

The aim of the study was to establish the complete microbiological profile of boar semen (Sus scrofa domesticus) and to choose the most effective antiseptic measures in order to control and optimize AI reproduction in pig farms. One hundred and one semen samples were collected and analyzed from several pig farms. The microbiological profile of ejaculates was determined by evaluating the degree of contamination of fresh semen and after dilution with specific extenders. The bacterial and fungal load of fresh boar semen recorded an average value of 82.41/0.149 × 10³ CFU/mL, while after diluting the ejaculates the contamination value was 0.354/0.140 × 10³ CFU/mL. Twenty-four germs (15 bacterial and 9 fungal species) were isolated, the most common being Candida parapsilosis/sake (92%) and Escherichia coli (81.2%). Modification of the sperm collection protocol (HPBC) reduced contamination in raw sperm by 49.85% in bacteria (significant (p < 0.00001) and by 9.67% in fungi (non-significant (p < 0.111491). The load in bacteria and filamentous fungi can be controllable, but not in levuras fungi. Some fluconazole-added extenders (12.5 mg%), ensure fungal aseptization, and even an increase in sperm progressivity (8.39%) for at least a 12 h shelf life after dilution. Validation of sperm aseptization was done by maintaining sow fecundity unchanged after AI (insignificant p > 0.05).     

Antimicrobial Properties of Lyophilized Extracts of Olive Fruit,Pomegranate and Orange Peel Extracts against Foodborne Pathogenic and Spoilage Bacteria and Fungi In Vitro and in Food Matrices

Several novel antimicrobials with different concentrations of olive, pomegranate, and orange fruit pulp extracts were produced from agricultural byproducts and, after lyophilization, their antimicrobial activity and potential synergistic effects were evaluated in vitro and in food samples against foodborne pathogenic and spoilage bacteria and fungi. The Minimum Inhibitory of the tested bacteria was 7.5% or 10%, while fungi were inhibited at a concentration of 10% or above. The optical density of bacterial and yeast cultures was reduced to a different extent with all tested antimicrobial powders, compared to a control without antimicrobials, and mycelium growth of fungi was also restricted with extracts containing at least 90% olive extract. In food samples with inoculated pathogens and spoilage bacteria and fungi, the 100% olive extract was most inhibitory against E. coli, S. typhimurium, and L. monocytogenes in fresh burger and cheese spread samples (by 0.6 to 1.8 log cfu/g), except that S. typhimurium was better inhibited by a 90% olive and 10% pomegranate extract in burgers. The latter extract was also the most effective in controlling the growth of inoculated fungi (Aspergillus niger, Penicillium italicum, Rhodotorula mucilaginosa) in both yogurt and tomato juice samples, where it reduced fungal growth by 1–2.2 log cfu/g at the end of storage period. The results demonstrate that these novel encapsulated extracts could serve as natural antimicrobials of wide spectrum, in order to replace synthetic preservatives in foods and cosmetics.     

Hemolytic and Antimicrobial Activities Differ Among Saponin-rich Extracts From Guar, Quillaja, Yucca, and Soybean

Hemolytic and antibacterial activities of eight serial concentrations ranged from 5-666 µg/mL of saponin-rich extracts from guar meal (GM), quillaja, yucca, and soybean were tested in 96-well plates and read by enzyme-linked immunosorbent assay plate-well as 650 nm. Hemolytic assay used a 1% suspension of chicken red blood cells with water and phosphate buffered saline as positive and negative controls, respectively. Antibacterial activity against Staphylococcus aureus, Salmonella typhimurium, and Escherichia coli were evaluated using ampicillin and bacteria without saponin-rich extract as positive and negative controls, respectively. The 100% MeOH GM and commercial quillaja saponin-rich extracts were significantly the highest in both hemolytic and antibacterial activities against all bacteria at the same concentration tested. Soybean saponin-rich extract had no antibacterial activity against any of the bacteria at the concentrations tested while yucca saponin-rich extract had no antibacterial activity against the gram-negative bacteria at the concentrations tested. GM and quillaja saponin-rich extracts were hemolytic, while yucca and soybean saponin-rich extracts were not hemolytic at the concentrations tested. No saponin-rich extract source had antibacterial activity against S. typhimurium or E. coli at the concentrations tested. Both GM and quillaja saponin-rich extracts exhibited antibacterial activity against S. aureus. Saponin-rich extracts from different plant sources have different hemolytic and antibacterial activities.     

Composition, antioxidant, antimicrobial and anti-wood-decay fungal activities of the twig essential oil of Taiwania cryptomerioides from Taiwan

This study investigated the chemical composition, antioxidant, antimicrobial and anti-wood-decay fungal activities of the essential oil isolated from the twigs of Taiwania cryptomerioides from Taiwan. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. A total of 35 compounds were identified, representing 100% of the oil. The main components identified were alpha-cadinol (45.9%), ferruginol (18.9%) and beta-eudesmol (10.8%). The antioxidant activity of the oil was tested by the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging capability test. The results showed an IC50 of 90.8 +/- 0.2 microg/mL. The active source compound was ferruginol. The antimicrobial activity of the oil was tested by the disc diffusion and micro-broth dilution methods against ten microbial species. The oil exhibited strong growth suppression against Gram-positive bacteria and yeast with inhibition zones of 45-52 mm and MIC values of 31.25-62.5 microg/mL, respectively. The anti-wood-decay fungal activity of the oil was also evaluated. The oil demonstrated excellent activity against four wood-decay-fungal species. For the antimicrobial and anti-wood-decay fungal activities of the oil, the active source compounds were determined to be alpha-cadinol, beta-eudesmol and ferruginol.     

Expression Analysis of Rice Pathogenesis-related Proteins Involved in Stress Response and Endophytic Colonization Properties of gfp-tagged Bacillus subtilis CB-R05

Bacillus subtilis CB-R05, possessing antagonistic effects against several fungal pathogens, is a diazotrophic plant growth-promoting bacteria marked with the green fluorescent protein (gfp) gene. To confirm the expression level of the pathogenesis-related (PR) proteins in rice inoculated with CB-R05, the expressions of four pathogenesis-related (PR) proteins (PR2, PR6, PR15, and PR16) were examined in the rice leaves treated with wounding stress over a time period. The PR proteins were generally more strongly expressed in the rice leaves inoculated with CB-R05 compared with the untreated control. The marked gfp-tagged B. subtilis CB-R05 strain was inoculated onto the rice seedlings under axenic conditions. Under the confocal laser scanning microscope (CLSM), the gfp-tagged CB-R05 bacterial cells were observed to penetrate the rhizoplane, especially in the elongation and differentiation zones of the rice roots, and colonize the root intracellularly. The bacteria, 24 h after the gfp-tagged CB-R05 inoculation, were seen to penetrate into the cell wall, cortex, xylem, and concentrate mainly in the vascular bundle. Numerous bacteria were observed within the intercellular spaces, root cortical cells, and xylem vessels. Over time, these bacteria dispersed to the lateral root junctions and propagated slowly from the roots to the stems and leaves. The B. subtilis CB-R05 population in the rice root rhizosphere was also monitored. These results show a very widespread colonization of the B. subtilis CB-R05 in the rice rhizosphere. Further attempts are under way to investigate the competition between the CB-R05 bacteria and the fungal pathogen in vivo.     

Voice Prosthesis Coated with Sustained Release Varnish Containing Clotrimazole Shows Long-Term Protection against Candida albicans: An In Vitro Study

Fungal biofilm formation on voice prosthesis (VP) is a major health problem that requires repeated replacement of the prosthesis. Candida albicans is one of the pathogens that frequently inhabits the VP. We proposed that coating VPs with sustained-release varnish (SRV) containing clotrimazole (CTZ) might prevent fungal biofilm formation. The long-term antifungal activities of SRV-CTZ- versus SRV-placebo-coated VPs was tested daily by measuring the inhibition zone of C. albicans seeded on agar plates or by measuring the fungal viability of C. albicans in suspension. The extent of biofilm formation on coated VPs was analyzed by confocal microscopy and scanning electron microscopy. We observed that SRV-CTZ-coated VPs formed a significant bacterial inhibition zone around the VPs and prevented the growth of C. albicans in suspension during the entire testing period of 60 days. Fungal biofilms were formed on placebo-coated VPs, while no significant biofilms were observed on SRV-CTZ-coated VPs. HPLC analysis shows that CTZ is continuously released during the whole test period of 60 days at a concentration above the minimal fungistatic concentration. In conclusion, coating VPs with an SRV-CTZ film is a potential effective method for prevention of fungal infections and biofilm formation on VPs.     

Piper betle (L): Recent Review of Antibacterial and Antifungal Properties,Safety Profiles,and Commercial Applications

Piper betle (L) is a popular medicinal plant in Asia. Plant leaves have been used as a traditional medicine to treat various health conditions. It is highly abundant and inexpensive, therefore promoting further research and industrialization development, including in the food and pharmaceutical industries. Articles published from 2010 to 2020 were reviewed in detail to show recent updates on the antibacterial and antifungal properties of betel leaves. This current review showed that betel leaves extract, essential oil, preparations, and isolates could inhibit microbial growth and kill various Gram-negative and Gram-positive bacteria as well as fungal species, including those that are multidrug-resistant and cause serious infectious diseases. P. betle leaves displayed high efficiency on Gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa, Gram-positive bacteria such as Staphylococcus aureus, and Candida albicans. The ratio of MBC/MIC indicated bactericidal and bacteriostatic effects of P. betle leaves, while MFC/MIC values showed fungicidal and fungistatic effects. This review also provides a list of phytochemical compounds in betel leaves extracts and essential oils, safety profiles, and value-added products of betel leaves. Some studies also showed that the combination of betel leaves extract and essential oil with antibiotics (streptomycin, chloramphenicol and gentamicin) could provide potentiating antibacterial properties. Moreover, this review delivers a scientific resume for researchers in respected areas and manufacturers who want to develop betel leaves-based products.     

Design,synthesis and SAR study of novel sulfonylurea derivatives containing arylpyrimidine moieties as potential anti-phytopathogenic fungal agents

Three series of novel sulfonylureas (SUs) 9-11 containing aromatic-substituted pyrimidines were designed and synthesized. The 3D-QASR and molecular docking studies showed that SUs should be considered as potential antiphytopathogenic fungal agents.     

Design,synthesis and SAR study of novel sulfonylurea derivatives containing arylpyrimidine moieties as potential anti-phytopathogenic fungal agents

Acetohydroxyacid synthase(AHAS) was considered as a promising target for antifungal agents.Herein,three series of novel sulfonylureas(SUs) 9-11 containing aromatic-substituted pyrimidines were designed and synthesized according to pharmacophore-combination and bioisosterism strategy.The in vitro fungicidal activities against ten phytopathogenic fungi indicated that most of the title compounds exhibited broad-spectrum and excellent fungicidal activities.Based on the preliminary fungicidal activities,a CoMFA model was constructed and the 3 D-QSAR analysis indicated that either a bulky group around the 5-position of the pyrimidine ring or electropositive group around the 2-position of the benzene ring would be favour to fungicidal activities.In order to study interaction mechanism,10 k was automatically docked into yeast AHAS and it further indicated that bearing bulky groups-aryl at the pyrimidine ring was critical to enhance antifungal activities.It revealed that the antifungal activity of derivatives 9-11 probably results from the inhibition of fungal AHAS.Thus,the present results strongly showed that SUs should be considered as lead compounds or model molecules to develop novel antiphyt o pathogenic fungal agents.     

Evaluation of bactericidal and fungicidal activity of ferrocenyl or phenyl derivatives in the diphenyl butene series

Some ferrocene compounds, such as tamoxifen derivatives hydroxyferrocifen 1 and ferrociphenol 2, show strong antiproliferative activity on hormone-dependent and hormone-independent breast cancer cells. In order to evaluate their antimicrobial activity, they were tested, together with their purely organic analogs, on the bacteria Pseudomonas aeruginosa and Staphylococcus aureus and the fungus Candida albicans. It has been found that the compounds bearing alkylamino chains are active, and in these cases the antimicrobial activity increases for compounds bearing two amino chains. These dialkyamino compounds are equally as active as doxycycline on P. aeruginosa and S. aureus but superior to it on C. albicans. The results show that there are no general correlation between the antitumoral activity and the bactericidal and fungicidal activities of these compounds. The ferrocene derivatives and their organic analogs have similar activity on bacteria and fungus. This bactericidal and fungicidal behaviour is a novel area of activity for these entities.     

Interaction of biomass of aerobic bacteria and fungi with Pu(IV) at low pH

The aim of this study was to investigate the change of Pu oxidation states due to interaction with aerobic bacteria and fungi at low pH under laboratory conditions. Microorganisms were isolated from samples collected from the low-level radioactive waste repository within the confines of Ignalina NPP. Abilities of the fungi (Absidia spinosa var spinosa Lendn. and Paecilomyces lilacinus Thom Samson) as well as Gram-positive bacteria (Bacillus mycoides and Micrococcus luteus) and Gram-negative bacterium I-m1 to transform the oxidation states of Pu under aerobic conditions were investigated. Oxidized and reduced Pu was tested using two radiochemical procedures. The amount of reduced and oxidized Pu was determined by measuring alpha activity after radiochemical separation. The results have shown that all bacteria and fungi can very slightly alter oxidation states of Pu due to their microbial activity. All the microorganisms tested demonstrate quite a fast process of Pu biosorption under the experimental conditions.     

Antimicrobial activity of three baccharis species used in the traditional medicine of Northern Chile

The antimicrobial activities of aqueous ethanol and chloroform extracts of three Baccharis species currently used in Northern Chile folk medicine for the treatment of several infectious and inflammatory disorders were tested against Gram-positive and negative bacteria and fungal spp. using the agar-disc diffusion assay. The results indicated that the activity was more pronounced against Gram-positive than against Gram-negative bacteria and yeast. No significant differences on the antibacterial activity were observed in the aqueous ethanol versus chloroform extracts. None of the plant extracts evaluated exhibited any activity against ten fungi tested.     

Phyllostictines A-D, oxazatricycloalkenones produced by Phyllosticta cirsii, a potential mycoherbicide for Cirsium arvense biocontrol

Phyllosticta cirsii, a fungal pathogen isolated from Cirsium arvense and proposed as biocontrol agent of this noxious perennial weed, produces in liquid cultures different phytotoxic metabolites with potential herbicidal activity. Four new oxazatricycloalkenones, named phyllostictines A-D, were isolated and characterized using essentially spectroscopic and chemical methods. Tested by leaf-puncture assay on the fungal host plant phyllostictine A proved to be highly toxic. The phytotoxicity decreases when both the dimension and the conformational freedom of the macrocyclic ring change, as in phyllostictines B and D, and it is totally lost when also the functionalization of the same ring is modified, as in phyllostictine C. Beside its phytotoxic properties, phyllostictine A has no antifungal activity, an interesting antibiotic activity only against Gram+ bacteria, and a noticeable zootoxic activity when tested at high concentrations. The integrity of the oxazatricycloalkenone system appears to be an important feature to preserve these activities.