The influence of chlorination timing and concentration on microbial communities in labyrinth channels: implications for biofilm removal

Chlorination is an effective method to control biofilm formation in enclosed pipelines. To date, very little is known about how to control biofilms at the mesoscale in complex pipelines through chlorination. In this study, the dynamic of microbial communities was examined under different residual chlorine concentrations on the biofilms attached to labyrinth channels for drip irrigation using reclaimed water. The results indicated that the microbial phospholipid fatty acids, extracellular polymeric substances, microbial dynamics, and the ace and Shannon microbial diversity indices showed a gradual decrease after chlorination. However, chlorination increased microbial activity by 0.5–19.2%. The increase in the relative abundances of chloride-resistant bacteria (Acinetobacter and Thermomonas) could lead to a potential risk of chlorine resistance. Thus, keeping a low chlorine concentration (0.83?mg l^?1 for 3?h) is effective for controlling biofilm formation in the labyrinth channels.     

Competitive interactions in mixed-species biofilms containing the marine bacterium Pseudoalteromonas tunicata

Pseudoalteromonas tunicata is a biofilm-forming marine bacterium that is often found in association with the surface of eukaryotic organisms. It produces a range of extracellular inhibitory compounds, including an antibacterial protein (AlpP) thought to be beneficial for P. tunicata during competition for space and nutrients on surfaces. As part of our studies on the interactions between P. tunicata and the epiphytic bacterial community on the marine plant Ulva lactuca, we investigated the hypothesis that P. tunicata is a superior competitor compared with other bacteria isolated from the plant. A number of U. lactuca bacterial isolates were (i) identified by 16S rRNA gene sequencing, (ii) characterized for the production of or sensitivity to extracellular antibacterial proteins, and (iii) labeled with a fluorescent color tag (either the red fluorescent protein DsRed or green fluorescent protein). We then grew single- and mixed-species bacterial biofilms containing P. tunicata in glass flow cell reactors. In pure culture, all the marine isolates formed biofilms containing microcolony structures within 72 h. However, in mixed-species biofilms, P. tunicata removed the competing strain unless its competitor was relatively insensitive to AlpP (Pseudoalteromonas gracilis) or produced strong inhibitory activity against P. tunicata (Roseobacter gallaeciensis). Moreover, biofilm studies conducted with an AlpP- mutant of P. tunicata indicated that the mutant was less competitive when it was introduced into preestablished biofilms, suggesting that AlpP has a role during competitive biofilm formation. When single-species biofilms were allowed to form microcolonies before the introduction of a competitor, these microcolonies coexisted with P. tunicata for extended periods of time before they were removed. Two marine bacteria (R. gallaeciensis and P. tunicata) were superior competitors in this study. Our data suggest that this dominance can be attributed to the ability of these organisms to rapidly form microcolonies and their ability to produce extracellular antibacterial compounds.     

The exo-proteome and exo-metabolome of Nostoc punctiforme (Cyanobacteria) in the presence and absence of nitrate

The ability of nitrogen-fixing filamentous Cyanobacteria to adapt to multiple environments comes in part from assessing and responding to external stimuli, an event that is initiated in the extracellular milieu. While it is known that these organisms produce numerous extracellular substances, little work has been done to characterize both the metabolites and proteins present under standard laboratory growth conditions. We have assessed the extracellular milieu of Nostoc punctiforme when grown in liquid culture in the presence and absence of a nitrogen source (nitrate). The extracellular proteins identified were enriched in integrin β-propellor domains and calcium-binding sites with sequences unique to N. punctiforme, supporting a role for extracellular proteins in modulating species-specific recognition and behavior processes. Extracellular proteases are present and active under both conditions, with the cells grown with nitrate having a higher activity when normalized to chlorophyll levels. The released metabolites are enriched in peptidoglycan-derived tetrasaccharides, with higher levels in nitrate-free media.     

Allelopathy in tropical and subtropical species

The ability of certain plants to synthesize allelochemicals that disrupt the germination, development, reproduction and/or survival of organisms that compete with them for resources has been observed in a variety of environments worldwide. Tropical and subtropical regions are particularly conducive to the evolution of allelopathic survival strategies as the relatively constant temperatures and mild frost-free winters produce a hospitable year-round growing season. This allows for the proliferation of a large variety of species and leads to fierce competition for sunlight, nutrients, water and other resources. Allelopathy provides an advantage to invasive species allowing for increased competitiveness and fitness over native and agricultural species in a variety of different habitats. Herein, the diversity and known action mechanisms of allelopathic compounds with a focus on tropical and subtropical communities is reviewed. Furthermore, the current and future prospect of utilizing and developing these allelopathic chemicals as weed control options is discussed.     

Environmental Control of Vanadium Haloperoxidases and Halocarbon Emissions in Macroalgae

Vanadium-dependent haloperoxidases (V-HPO), able to catalyze the reaction of halide ions (Cl^?, Br^?, I^-) with hydrogen peroxide, have a great influence on the production of halocarbons, which in turn are involved in atmospheric ozone destruction and global warming. The production of these haloperoxidases in macroalgae is influenced by changes in the surrounding environment. The first reported vanadium bromoperoxidase was discovered 40 years ago in the brown alga Ascophyllum nodosum. Since that discovery, more studies have been conducted on the structure and mechanism of the enzyme, mainly focused on three types of V-HPO, the chloro- and bromoperoxidases and, more recently, the iodoperoxidase. Since aspects of environmental regulation of haloperoxidases are less well known, the present paper will focus on reviewing the factors which influence the production of these enzymes in macroalgae, particularly their interactions with reactive oxygen species (ROS).     

De novo formation of organochlorines in a sewage treatment plant

The de novo formation of organochlorines was observed in a municipalsewage treatment plant. Due to this formation, the amount of organically boundhalogens (AOX) increased 15-fold inside the sewage treatment plant. Per day,more than 6 kg of organically-bound chlorine were produced. Thisformation is not based on a metabolism of present organochlorines, it is a denovo formation out of inorganic chloride and organic substrates. The AOXtriggerconcentration in sewage sludge in Germany is 500 mgkg^–1 and was sometimes exceeded by a factor of 10. Noknown anthropogenic organohalogens were found which could explain the elevatedAOX concentrations. Instead many chlorinated compounds could be identifiedwhichwere not known to be of anthropogenic origin. The compound with the highestconcentration was the 3,4-dichlorophenylacetic acid (3,4-CPAc). In one case,more than 1 g kg^–1 of this compound was detected.A slaughterhouse that emits phenylacetic acid is probably the origin of thatformation. In model experiments phenylacetic acid was chlorinated with HOCl butchlorinated phenylacetic acids other than 3,4-CPAc were found. Therefore it canbe excluded that the chlorination in the sewage treatment plant takes place byan abiotic reaction with hypochlorite that might have been introduced there. Weassume that the occurring microorganisms are responsible for the de novoformation in the sewage treatment plant. The obtained knowledge could also beuseful to understand natural chlorination processes.     

Plant metabolomics for plant chemical responses to belowground community change by climate change

General circulation models on global climate change predict increase in surface air temperature and changes in precipitation. Increases in air temperature (thus soil temperature) and altered precipitation are known to affect the species composition and function of soil microbial communities. Plant roots interact with diverse soil organisms such as bacteria, protozoa, fungi, nematodes, annelids and insects. Soil organisms show diverse interactions with plants (eg. competition, mutualism and parasitism) that may alter plant metabolism. Besides plant roots, various soil microbes such as bacteria and fungi can produce volatile organic compounds (VOCs), which can serve as infochemicals among soil organisms and plant roots. While the effects of climate change are likely to alter both soil communities and plant metabolism, it is equally probable that these changes will have cascading consequnces for grazers and subsequent food web components aboveground. Advances in plant metabolomics have made it possibile to track changes in plant metabolomes as they respond to biotic and abiotic environmental changes. Recent developments in analytical instrumentation and bioinformatics software have established metabolomics as an important research tool for studying ecological interactions between plants and other organisms. In this review, we will first summarize recent progress in plant metabolomics methodology and subsequently review recent studies of interactions between plants and soil organisms in relation to climate change issues.     

Mutagenicity on chlorination of products formed by ozonation of naphthoresorcinol in water

The mutagenicity of products formed by chlorination after ozonation of naphthoresorcinol in aqueous solution was assayed with Salmonella typhimurium strains TA98 and TA100 in the presence and absence of S9 mix from phenobarbital- and 5,6-benzoflavone-induced rat liver. Ozonated and subsequently chlorinated naphthoresorcinol was directly mutagenic, as was ozonated naphthoresorcinol, in both strains tested. The mutagenic activity at chlorination with 8 equivalents of chlorine per mole of naphthoresorcinol after ozonation was markedly higher than that at only ozonation. Of the identified ozonation products of naphthoresorcinol, muconic acid, after chlorination with 2 or 4 equivalents of chlorine per mole of the compound, induced direct mutagenicity against TA98 and TA100. The chlorination of glyoxal with 0.5 and 1 chlorine equivalents per mole of the compound was shown to produce direct mutagenicity toward TA98. The identification of the chlorination products of these compounds is also discussed.     

Extracellular Deoxyribonuclease Production by Yeasts

A total of 20 genera of yeasts and yeastlike organisms were tested for their ability to produce an extracellular deoxyribonuclease. Results indicate that ability to produce the enzyme appears to be a specific characteristic of the three genera Rhodotorula, Cryptococcus, and Tremella. A single strain of Endomycopsis fibuligera was also shown to be positive for the enzyme. In comparing the ability of the organisms to excrete extracellular deoxyribonuclease with their ability to produce urease, a surprisingly close correlation was found. With the exception of Lipomyces starkeyi, all the organisms which were deoxyribonuclease-negative were also urease-negative. Of those organisms which were deoxyribonuclease-positive, only E. fibuligera was urease-negative. The ability of cryptococci to produce extracellular deoxyribonuclease is discussed in relation to the implication which this finding may have for the taxonomy and phylogeny of the genus.     

Interactions of arboreal yeast endophytes: an unexplored discipline

The value of healthy forest ecosystems is well known and trees in these systems form symbioses with a variety of living organisms. This review focuses on literature pertaining to the potential interactions of arboreal yeast endophytes with trees and their associated insects. Although very little is known about the symbioses of arboreal yeast endophytes, indications are that some of these unicellular fungi produce plant-growth promoting phytohormones, while others are antagonistic towards phytopathogens or are capable of producing pheromones that affect the behavior of insect herbivores. However, more research needs to be conducted to fully understand the role of arboreal yeast endophytes in ecosystem processes.     

Isolation and characterization of protease overproducing mutants of Trichoderma harzianum

Several Trichoderma strains have been reported to be effective in controlling plant diseases, and the action of fungal hydrolytic enzymes is considered as the main mechanism involved in the antagonistic process. Strain Trichoderma harzianum T334 is a potential biocontrol agent against plant pathogenic fungi with the ability to produce low levels of proteases constitutively. To improve its fungal antagonistic capacity, mutagenetic program was undertaken for the construction of protease overproducing derivates. The mutant strains were obtained by means of UV-irradiation and were selected for p-fluorophenyl-alanine resistance or altered colony morphology. It was revealed by means of specific chromogenic protease substrates that both trypsin-like and chymotrypsin-like protease secretion was elevated in most of the mutant strains. The profiles of isoenzymes were different between the mutants and the wild-type strain, when examined by gel filtration chromatography. Certain mutants proved to be better antagonists against plant pathogens in in vitro antagonism experiments. This study suggests the possibility of using mutants with improved constitutive extracellular protease secretion against plant pathogenic fungi.     

Attachment as a factor in the protection of Enterobacter cloacae from chlorination

Enterobacter cloacae attached to drinking water distribution particles was subjected to chlorination. Attachment resulted in the protection of these organisms from disinfection. This effect was found to be dependent upon both the level of chlorine in the system and attachment time. The results obtained in this study indicate that attached organisms may play an important role in coliform outbreaks.     

Attachment as a factor in the protection of Enterobacter cloacae from chlorination.

Enterobacter cloacae attached to drinking water distribution particles was subjected to chlorination. Attachment resulted in the protection of these organisms from disinfection. This effect was found to be dependent upon both the level of chlorine in the system and attachment time. The results obtained in this study indicate that attached organisms may play an important role in coliform outbreaks.     

Effects of previous nitrogen addition on chlorine in forest soil, soil solution and biomass

There is increasing evidence that forests and forest soil contribute to the signature of chlorine composition in water bodies. However, little is known about the potential effects of land management activities on chlorine biogeochemistry. This study examines the effects of previous nitrogen addition on chlorine chemistry in a Pinus sylvestris L. forest located in south-central Sweden (60°00′N, 13°43′E). Repeated addition of nitrogen to study plots over a 20-year period resulted in total additions of 0, 450 and 900 kg N ha^?1. Soil samples were collected before harvesting, and soil solution and biomass were sampled following final felling. Contrary to previous findings, we found no clear evidence that previous addition of nitrogen had hampered the formation of organic chlorine in the organic soil layer. We suggest that the effects of nitrogen addition on chlorination processes are not seen in the surface soil, but are instead manifested in the migration of organic matter in the mineral soil. Soil organic matter from the E-horizon had a lower chlorination degree in the nitrogen-amended plots than in the control plots. In addition, we observed lower Cl^? levels in the seedling needles following high nitrogen fertilization (900 N) than in 450 N or control. These results add on the importance of studying chlorine dynamics with focus on what chlorinated soil organic matter can be resistant to degradation compared to average soil organic matter and what can be leached as a result of harvesting and available for vegetation.     

Plant extracellular ATP signalling: new insight from proteomics

Complex signalling systems have evolved in multicellular organisms to enable cell-to-cell communication during growth and development. In plants, cell communication via the extracellular matrix (apoplast) controls many processes vital for plant survival. Secretion of ATP into the extracellular matrix is now recognised as a previously unknown stimulus for cell signalling with a role in many aspects of plant physiology. In the last decade, the secondary messenger molecules in extracellular ATP signalling were identified, but the downstream gene and protein networks that underpin plant responses to extracellular ATP are only beginning to be characterised. Here we review the current status of our knowledge of plant extracellular signalling and demonstrate how applying state-of-the art proteomic technologies is rapidly bringing new discoveries in extracellular ATP research. We discuss how monitoring of the global proteomic profile during responses to modulation of extracellular ATP signalling has led to novel insight into pathogen defence systems and plant programmed cell death regulation. On the basis of extensive proteomic, pharmacological, and reverse genetics data, extracellular ATP has been confirmed to constitute an important molecular switch that tightly controls organellar energy metabolism, reprogramming of primary metabolic pathways, and redirection of resources to protein networks that support adaptation of plants to stress.     

Virulence plasmid-associated adhesion of Escherichia coli and its significance for chlorine resistance

Introduction of the ColV, I-K94 virulence plasmid into strains of Escherichia coli led (for four out of five strains tested) to a marked increase in the ability of organisms to adhere to glass beads. For strain 1829, the plasmid led to increased attachment to other materials including sand, agar, agarose, chitin and cellulose. The increased adhesion to glass beads was due to the presence of the plasmid and not to its introduction into a variant with altered adhesive properties. The plasmid-encoded VmpA protein did not appear to be necessary for the ColV, I-K94-promoted adhesion but adhesion was absolutely dependent on the presence of derepressed levels of transfer components in the ColV+ strains and partially dependent on the presence of colicin components. The extent of the plasmid-promoted adhesion was greatest for organisms grown at 30 degrees, 37 degrees or 42 decrees C and adhesion was almost abolished by growth at 21 degrees or 25 degrees C; this finding is in accord with transfer and colicin components being involved in adhesion. Of several other plasmids tested for their effects on adhesion, those with derepressed transfer properties showed a marked effect as did the RI resistance plasmid. Because of the ease of handling glass bead-attached organisms, such preparations were used as a model for studying the relevance of attachment to the resistance of E. coli to chlorination in the water purification process. Organisms of 1829 ColV, I-K94, attached to glass beads, were more resistant to damage and killing by chlorine than were unattached organisms. Three findings suggest that such chlorine resistance may be significant for survival during water chlorination. Firstly, ColV, I-K94+ bacteria became attached if incubated in sewage effluent with glass beads at 20 degrees C. Secondly, ColV+ organisms already attached to glass beads maintained their attachment during 24 h incubation in effluent at 20 degrees C and thirdly such effluent incubated organisms remained chlorine resistant provided that they retained their attachment.     

Small worms, big appetites: ratios of different functional morphs in relation to interspecific competition in trematode parasites

Animals living in colonies or collectives composed of highly-related individuals often produce morphs that are physically and behaviourally specialised to perform specific tasks. Because such morphs are often sterile, their production represents a fitness cost for the colony and there should be an optimal ratio of the numbers of sterile specialists and reproductive members that may be adjustable to environmental conditions. Trematode parasites undergo asexual multiplication within their snail intermediate host, resulting in large numbers of clonal stages known as rediae or sporocysts, depending on the trematode species. In areas with high prevalences of infection, the host can be infected by multiple species, which can lead to intense competition for limited resources. Here, we describe the existence of specialised ‘mini-rediae’ in the trematode Philophthalmus sp. that are morphologically and functionally specialised for interspecific competition. Mini-rediae were observed feeding on the sporocysts of a co-occurring trematode species - Maritrema novaezealandensis. In addition, in larger snails - which are less likely to have M. novaezealandensis infections - Philophthalmus sp. produces relatively fewer mini-rediae than expected. Our findings support results from a prior study which demonstrated the existence of morphs that perform specialised functions in antagonistic interspecific interactions in trematodes, and additionally shows that the number of these morphs in each host is associated with the likelihood of encountering other species within the same host. Trematodes may thus provide interesting models for studying morphological specialisation in colonial organisms.     

A recent advance in the intracellular and extracellular redox post‐translational modification of proteins in plants

Plants, as sessile organisms, have acquired through evolution sophisticated regulatory signal pathways to overcome external variable factors during each stage of the life cycle. Among these regulatory signals, two pathways in particular, reactive oxygen species and reactive nitrogen species, have become of significant interest in several aspects of plant biology, underpinning these molecules as critical regulators during development, cellular differentiation, and plant‐pathogen interaction. Recently, redox posttranslational modifications (PTM), such as S‐nitrosylation on cysteine residues and tyrosine nitration, have shed light on multiple protein targets, as they are associated with signal networks/downstream metabolic pathways, capable of transducing the imbalance of redox hemostasis and consequently redirecting the biochemical status under stress conditions. However, most of the redox PTM have been studied only in the intracellular compartment, providing limited information concerning redox PTM in the extracellular matrix of plant cells. Nevertheless, recent studies have indicated the plausibility of redox PTM in extracellular proteins, including cell wall associated proteins. Accordingly, in this review, we endeavor to examine evidence of redox PTM supported by mass spectrometry data in the intracellular and extracellular space in plant cells. As a further example, we focus the last section of this review on illustrating, using molecular dynamics simulation, the effect of S‐nitrosylation on the structural conformation of well‐known cell wall‐associated proteins including pectin methylesterase and xyloglucan endo‐transglycosylases.     

Halogenation and Oxidation Reactions with Haloperoxidases

Haloperoxidases are enzymes which catalyze the incorporation of halogen atoms into organic molecules. They are found throughout nature, playing a major role in the defence system of many organisms. Their reaction mechanisms as well as their use as catalysts for halogenation and oxidation reactions on laboratory and industrial scales are discussed. Up to now, selective halogenation reactions have only been reported for the chloroperoxidase from Pseudomonas pyrrocinia. The usefulness of the other enzymes is based on their ability to produce hypohalous acid (HOX) in a controllable way, allowing the smooth (yet nonselective) halogenation of electron-rich substrates. On the other hand, it has been shown recently that some haloperoxidases can stereoselectively convert sulfides and alkenes into their corresponding homochiral oxides. Therefore, these enzymes will undoubtedly gain importance in the near future.     

Fungal chitinases and their biological role in the antagonism onto nematode eggs. A review

Chitin, the most abundant aminopolysaccharide in nature, is a rigid and resistant structural component that contributes to the mechanical strength of chitin-containing organisms. Chemically, it is a linear cationic heteropolysaccharide composed of N-acetyl-D-glucosamine and D-glucosamine units. The enzymatic degradation of chitin is performed by a chitinolytic system with synergistic and consecutive action. Diverse organisms (containing chitin or not) produce a great variety of chitinolytic enzymes with different specificities and catalytic properties. Their physiological roles involve nutrition, parasitism, chitin recycling, morphogenesis, and/or defense. Microorganisms, as the main environmental chitin degraders, constitute a very important natural source of chitinolytic enzymes. Nowadays, the most used method for pest and plant diseases control is the utilization of chemical agents, causative of significant environmental pollution. Social concern has generated the search for alternative control systems (i.e., biological control), which contribute to the generation of sustainable agricultural development. Interactions among the different organisms are the natural bases of biological control. Interest in chitinolytic enzymes in the field of biological control has arisen due to their possible involvement in antagonistic activity against pathogenic chitin-containing organisms. The absence of chitin in plants and vertebrate animals allows the consideration of safe and selective “target” molecules for control of chitin-containing pathogenic organisms. Fungi show appropriate characteristics as potential biological control agents of insects, fungi, and nematodes due to the production of fungal enzymes with antagonistic action. The antagonistic interactions between fungi and plant nematode parasites are among the most studied experimental models because of the high economic relevance. Fungi which target nematodes are known as nematophagous fungi. The nematode egg is the only structural element where the presence of chitin has been demonstrated. In spite of being one of the most resistant biological structures, eggs are susceptible to being attacked by egg-parasitic fungi. A combination of physical and chemical phenomena result in their complete destruction. The contribution of fungal chitinases to the in vitro rupture of the eggshell confirms their role as a pathogenic factor. Chitinases have been produced by traditional fermentation methods, which have been improved by optimizing the culture conditions for industrial processes. Although wild-type microorganisms constitute an alternative source of chitinolytic enzymes, the advances in molecular biology are allowing the genetic transformation of fungi to obtain strains with high capability as biocontrol agents. Simultaneously, a better understanding of rhizosphere interactions, additional to the discovery of new molecular biology tools, will allow the choosing of better alternatives for the biological control of nematodes in order to achieve an integrated management of the soil ecosystem.