Save the crop: Photodynamic Inactivation of plant pathogens I: bacteria.

The ever growing world-population poses challenges concerning the need for more food free of pesticide residues. The most common means to control plant pathogens is through the application of pesticides, which raises concerns over safety for humans and the environment. Recently, Photodynamic Inactivation (PDI) of microorganisms using natural photosensitizers has shown itself to be a powerful tool to combat bacteria and fungi. This study investigates the efficacy of PDI against the Gram(+) bacterial plant pathogen Rhodococcus fascians and Gram(-) Xanthomonas axonopodis and Erwinia amylovora using two chlorin e6 derivatives as photosensitizers: anionic sodium magnesium chlorophyllin (Chl, approved as food additive E140) in combination with cell wall permeabilizing agents (Na2EDTA or Polyaspartic acid sodium salt (PA)) and B17-0024, a mixture of chlorin e6 derivatives with cationic moieties at physiological pH. Both photosensitizers show excellent efficacy against R. fascians, whereby B17-0024 is phototoxic at a one order of magnitude lower concentration than Chl (10 µM B17-0024: relative inactivation (r.i.) >7.5 × 106, 100 µM Chl: r.i. 2.2 × 106, illumination with 26.6 J cm-2, 395 nm). The phototreatment of Gram(-) bacteria with Chl requires the obligatory use of cell wall permeabilizing agents like Na2EDTA (X. axonopodis) or PA (E. amylovora) to induce significant killing (more than 7 log units at 100 µM). On the other hand, B17-0024 proves to be a highly effective photosensitizer inducing bacterial inactivation at very low concentrations (10 µM for R. fascians and X. axonopodis, 100 µM for E. amylovora) without additives. In summary, PDI using both the natural photosensitizer Chl in combination with cell wall permeabilizing agents is effective and environmentally friendly. As an alternative, B17-0024 is highly photoactive against all model strains tested - even without cell wall permeabilizing agents. The photodynamic approach based on chlorin e6 derivatives should add to the growers' toolbox as a preferred alternative for the control of phytopathogens.

Lipid storage in high-altitude Andean Lakes extremophiles and its mobilization under stress conditions in Rhodococcus sp. A5, a UV-resistant actinobacterium.

The production of triacylglycerols (TAG) or wax esters (WS) seems to be a widespread feature among extremophile bacteria living in high-altitude Andean Lakes (HAAL), Argentina. Twelve out of twenty bacterial strains isolated from HAAL were able to produce TAG or WS (between 2 and 17 % of cellular dry weight) under nitrogen-limiting culture conditions. Among these strains, the extremophile Rhodococcus sp. A5 accumulated significant amounts of TAG during growth on glucose (17 %, CDW) and hexadecane (32 %, CDW) as sole carbon sources. The role of accumulated TAG in the response to carbon starvation, osmotic stress, UV-radiation and desiccation was investigated in Rhodococcus sp. A5 using an inhibitor of TAG degradation. Cells degraded TAG during these stresses in the absence of the inhibitor. The inhibition of TAG mobilization affected cell survival during osmotic stress only during the initial growth stage. Little or no surviving cells were observed after carbon starvation, UV-treatment and desiccation, when TAG mobilization was inhibited. These results suggested that TAG metabolism is relevant for the adaptation and survival of A5 cells under carbon starvation, osmotic stress and UV irradiation, and essential under desiccation conditions, which prevail in HAAL environments.

In vivo 31P and 13C NMR investigations of Rhodococcus rhodochrous metabolism and behaviour during biotransformation processes.

AIMS: The strain Rhodococcus rhodochrous OBT18 was isolated from a water treatment plant used to decontaminate industrial effluents containing benzothiazole derivatives. Aims of the work are to study the central metabolism of this strain and more specifically its behaviour during biodegradation of 2-aminobenzothiazole. METHODS AND RESULTS: In vivo(13)C and (31)P NMR experiments showed that this strain contains storage compounds such as polyphosphates, glycogen and trehalose and produces biosurfactants containing trehalose as sugar unit. Trehalose can be synthesized after reversion of the glycolytic pathway. In vivo(31)P NMR experiments showed that energy metabolism markers such as the intracellular pH and the ATP concentration did not change during biotransformation processes when R. rhodochrous was exposed to potentially toxic compounds including iron complexes and (* )OH radicals. Also R. rhodochrous recovers the normal values of ATP and pH after anoxia/reoxygenation cycle very quickly. CONCLUSIONS: Rhodococcus rhodochrous carbon and energy metabolism is well adapted to different stresses and consequently to live in the environment where conditions are constantly changing. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study can be used to understand the behaviour of this bacterium in natural environments but also in water treatment plants where iron and UV light are present.

Ultrasound assisted biodesulfurization of liquid fuel using free and immobilized cells of Rhodococcus rhodochrous MTCC 3552: A mechanistic investigation.

This paper attempts to gain mechanistic insight into enhancement effect of sonication on biodesulfurization. The approach has been to fit Haldane kinetics model to dibenzothiophene (DBT) metabolism and analyze trends in model parameters concurrently with simulations of cavitation bubble dynamics. Mechanistic synergy between sonication and biodesulfurization is revealed to be of both physical and chemical nature. Generation of micro-turbulence in medium by sonication leads to fine emulsification and enhancement of DBT transport across organic/aqueous interphase. Microturbulence also enhances transport of substrate and product across cell wall that increases reaction velocity (Vmax). Michaelis constant (Km) and inhibition constant (KI), being intrinsic parameters, remain unaffected by sonication. Radicals produced by transient cavitation oxidize DBT to DBT-sulfoxide and DBT-sulfone (intermediates of metabolism), which contributes enhancement of biodesulfurization. However, high shear generated by ultrasound and cavitation has adverse effect on action of surfactant ß-cyclodextrin for enhancement of interphase transport of DBT.

Effect of gamma radiation on native endolithic microorganisms from a radioactive waste deposit site.

A time-course experiment was conducted to evaluate the effects of gamma radiation on the indigenous microbiota present in rock obtained from Yucca Mountain, Nevada Test Site. Microcosms were constructed by placing pulverized Yucca Mountain rock in polystyrene cylinders. Continuous exposure (96 h) at a dose rate of 1.63 Gy/min was used to mimic the near-field environment surrounding waste canisters. The expected maximum surface dose rate from one unbreached canister designed to contain spent nuclear fuels is 0.06 Gy/min. Considering the current repository packing design, multiple canisters within one vault, the cumulative dose rate may well approach that used in this experiment. The microbial communities were characterized after receiving cumulative doses of 0, 0.098, 0. 58, 2.33, 4.67, 7.01 and 9.34 kGy. Radiation-resistant microorganisms in the pulverized rock became viable but nonculturable (VBNC) after a cumulative dose of 2.33 kGy. VBNC microorganisms lose the ability to grow on media on which they have routinely been cultured in response to the environmental stress imposed (i.e. radiation) but can be detected throughout the time course using direct fluorescence microscopy techniques. Two representative exopolysaccharide-producing isolates from Yucca Mountain were exposed to the same radiation regimen in sand microcosms. One isolate was much more radiation-resistant than the other, but both had greater resistance than the general microbial community based on culturable counts. However, when respiring cell counts (VBNC) were compared after irradiation, the results would indicate much more radiation resistance of the individual isolates and the microbial community in general. These results have significant implications for underground storage of nuclear waste as they indicate that indigenous microorganisms are capable of surviving gamma irradiation in a VBNC state.

Algae harvesting for biofuel production: influences of UV irradiation and polyethylenimine (PEI) coating on bacterial biocoagulation.

There is a pressing need to develop efficient and sustainable separation technologies to harvest algae for biofuel production. In this work, two bacterial species (Escherichia coli and Rhodococus sp.) were used as biocoagulants to harvest Chlorella zofingiensis and Scenedesmus dimorphus. The influences of UV irradiation and polyethylenimine (PEI)-coating on the algal harvesting efficiency were investigated. Results showed that the UV irradiation could slightly enhance bacteria-algae biocoagulation and algal harvesting efficiency. In contrast, the PEI-coated E. coli cells noticeably increased the harvesting efficiencies from 23% to 83% for S. dimorphus when compared to uncoated E. coli cells. Based on the soft-particle Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, an energy barrier existed between uncoated E. coli cells and algal cells, whereas the PEI coating on E. coli cells eliminated the energy barrier, thereby the biocoagulation was significantly improved. Overall, this work presented groundwork toward the potential use of bacterial biomass for algal harvesting from water.

Low-temperature ionizing radiation resistance of Deinococcus radiodurans and Antarctic Dry Valley bacteria.

The high flux of cosmic rays onto the unshielded surface of Mars poses a significant hazard to the survival of martian microbial life. Here, we determined the survival responses of several bacterial strains to ionizing radiation exposure while frozen at a low temperature characteristic of the martian near-subsurface. Novel psychrotolerant bacterial strains were isolated from the Antarctic Dry Valleys, an environmental analogue of the martian surface, and identified by 16S rRNA gene phylogeny as representatives of Brevundimonas, Rhodococcus, and Pseudomonas genera. These isolates, in addition to the known radioresistant extremophile Deinococcus radiodurans, were exposed to gamma rays while frozen on dry ice (-79°C). We found D. radiodurans to exhibit far greater radiation resistance when irradiated at -79°C than was observed in similar studies performed at higher temperatures. This greater radiation resistance has important implications for the estimation of potential survival times of microorganisms near the martian surface. Furthermore, the most radiation resistant of these Dry Valley isolates, Brevundimonas sp. MV.7, was found to show 99% 16S rRNA gene similarity to contaminant bacteria discovered in clean rooms at both Kennedy and Johnson Space Centers and so is of prime concern to efforts in the planetary protection of Mars from our lander probes. Results from this experimental irradiation, combined with previous radiation modeling, indicate that Brevundimonas sp. MV.7 emplaced only 30 cm deep in martian dust could survive the cosmic radiation for up to 100,000 years before suffering 106 population reduction.

Variación diurna en la composición del bacterioplancton y daño del ADN en la comunidad microbiana de un lago oligotrófico andino / Diurnal variation in bacterioplankton composition and DNA damage in the microbial community from an Andean oligotrophic lake

Laguna Azul is an oligotrophic lake situated at 4,560 m above sea level and subject to a high level of solar radiation. Bacterioplankton community composition (BCC) was analysed by denaturing gradient gel electrophoresis and the impact of solar ultraviolet radiation was assessed by measuring cyclobutane pyrimidine dimers (CPD). Furthermore, pure cultures of Acinetobacter johnsonii A2 and Rhodococcus sp. A5 were exposed simultaneously and CPD accumulation was studied. Gel analyses generated a total of 7 sequences belonging to Alpha-proteobacteria (1 band), Beta-proteobacteria (1 band), Bacteroidetes (2 bands), Actinobacteria (1 band), and Firmicutes (1 band). DGGE profiles showed minimal changes in BCC and no CPD was detected even though a high level of damage was found in biodosimeters. A. johnsonii A2 showed low level of DNA damage while Rhodococcus sp. A5 exhibited high resistance since no CPD were detected under natural UV-B exposure, suggesting that the bacterial community is well adapted to this highly solar irradiated environment.

La Laguna Azul es un ambiente oligotrófico localizado a 4560m de altura y sometido a elevados niveles de radiación solar. La composición de su comunidad bacterioplanctónica fue analizada empleando la técnica de electroforesis en gradiente desnaturalizante y se investigó el impacto de la radiación ultravioleta cuantificando los dímeros de pirimidina (CPD). Además, se expusieron simultáneamente cultivos puros de Acinetobacter johnsonii A2 y Rhodococcus sp. A5 para estudiar la acumulación de CPD. El análisis de los geles mostró siete secuencias pertenecientes a Alpha-proteobacteria (1 banda), Beta-proteobacteria (1 banda), Bacteroidetes (2 bandas), Actinobacteria (1 banda) y Firmicutes (1 banda). A lo largo del día se observaron cambios mínimos en la composición de la comunidad y no se detectaron CPD. A. johnsonii A2 presentó un daño bajo mientras que Rhodococcus sp. A5 no presentó daño en su ADN, sugiriendo que la comunidad bacteriana está muy bien adaptada a este ambiente altamente irradiado

Multiple-stress tolerance of ionizing radiation-resistant bacterial isolates obtained from various habitats: correlation between stresses.

Isolation of five ionizing radiation (IR)-resistant bacteria by screening of isolates from various habitats classified as common and stressed is reported. IR-resistant isolates exhibited varying degrees of resistance to gamma-radiation and were classified as highly and moderately radiation resistant. Resistance to ultraviolet (UV) radiation correlated well with gamma-radiation resistance, whereas a comparable desiccation resistance for all the highly and moderately radiation-resistant isolates was observed. However, salt tolerance failed to correlate with IR resistance, indicating a divergent evolution of the salt tolerance and radiation resistance. Characterization of isolates by the amplified rDNA restriction analysis profiling attested to the clustering of these isolates with their stress phenotype. 16S rRNA gene-based analysis of the isolates showed that the bacteria with similar-resistance physiologies clustered together and belonged to related genera. Hydrogen peroxide resistance and mitomycin survival patterns of the isolates indicated the roles of oxidative-stress tolerance in desiccation survival and recombination repair in higher radiation resistance, respectively.

[The effect of physicochemical environmental factors on the growth of gram-positive bacterial dissociants]. / Vliianie fiziko-khimicheskikh faktorov sredy na rost dissotsiantov nekotorykh grampolozhitel'nykh bakterii.

The growth of R-, S- and M (g)-dissociants of Streptococcus lactis, Bacillus coagulans, Rhodococcus rubropertinctus under the action of some physico-chemical factors: temperature, pH, ultraviolet (UV) rays, high concentration of NaCl and storage have been compared. R-variants gain selective advantage under the influence of UV-irradiation, high temperature and storage; S-variants--at decreasing of active pH of medium; M (g)-variants--at decreasing of growth temperature, high values of pH, increased NaCl concentration. The dissociation has been concluded to enlarge the limits of the species survival.