The Water Hyacinth Microbiome: Link Between Carbon Turnover and Nutrient Cycling.

Water hyacinth (WH), a large floating plant, plays an important role in the biogeochemistry and ecology of many freshwaters globally. Its biogeochemical impact on wetland functioning is strongly mediated by the microbiome associated with its roots. However, little is known about the structure and function of this WH rhizobiome and its relation to wetland ecosystem functioning. Here, we unveil the core and transient rhizobiomes of WH and their key biogeochemical functions in two of the world's largest wetlands: the Amazon and the Pantanal. WH hosts a highly diverse microbial community shaped by spatiotemporal changes. Proteobacteria lineages were most common, followed by Actinobacteria and Planctomycetes. Deltaproteobacteria and Sphingobacteriia predominated in the core microbiome, potentially associated with polysaccharide degradation and fermentation of plant-derived carbon. Conversely, a plethora of lineages were transient, including highly abundant Acinetobacter, Acidobacteria subgroup 6, and methanotrophs, thus assuring diverse taxonomic signatures in the two different wetlands. Our findings point out that methanogenesis is a key driver of, and proxy for, community structure, especially during seasonal plant decline. We provide ecologically relevant insights into the WH microbiome, which is a key element linking plant-associated carbon turnover with other biogeochemical fluxes in tropical wetlands.

Curvulin and spirostaphylotrichins R and U from extracts produced by two endophytic Bipolaris sp. associated to aquatic macrophytes with antileishmanial activity.

In the present study, biological activity and chemical composition of two crude extracts of endophytic fungal strains of Bipolaris genera isolated from two species of aquatic macrophytes: Eichhornia azurea (Kunth) and Eichhornia crassipes (Mart.) were investigated. The nuclear magnetic resonance and mass spectrometry data provided the identification of three main compounds: curvulin (1), spirostaphylotrichin R (2) and U (3). The fragmentation mechanism of the precursor ions towards collision induced dissociation (CID) tandem mass spectrometry experiment (MS/MS) is also proposed. Furthermore, biological screening of the crude extracts displayed antileishmanial activity with IC50 values ranging from 70-84.2 µg.mL-1.

Eichhornia azurea decomposition and the bacterial dynamic: an experimental research

Abstract Organic decomposition is a complex interaction between chemical, physical and biological processes, where the variety of aquatic vascular plants is essential for the trophic dynamics of freshwater ecosystems. The goal of this study was to determine the aquatic macrophyte Eichhornia azurea (Sw.) Kunth decomposition rate, the time relation with the limnological parameters, and whether this relationship is a result of decomposition processes. To that end, we collected water and leaves of E. azurea in Surf Leopoldo, PR. The experiment consisted of two treatments: 25 containers with 450 mL of water and 0.8 g of biomass dry weight were used with or without the addition of macrophytes. Samples were collected in triplicate at times 0, 3 h, 6 h, 12 h, 24 h, 72 h, 120 h, 168 h and 240 h. When the container was removed, the plant material was dried in an oven. After 48 h, the material was measured to obtain the final dry weight. Analyses of pH, conductivity, dissolved oxygen, total phosphorus N-ammonia (NH4), soluble reactive phosphorus (PO4) and dissolved organic carbon were performed, and the decomposition rate was calculated. The results showed significant temporal variation of limnological parameters in the study. Additionally, dissolved oxygen, conductivity, dissolved organic carbon and total phosphorus were correlated with the dry weight of the biomass, suggesting that E. azurea decomposition significantly interferes with the dynamics of these variables.

Eichhornia azurea decomposition and the bacterial dynamic: an experimental research.

Organic decomposition is a complex interaction between chemical, physical and biological processes, where the variety of aquatic vascular plants is essential for the trophic dynamics of freshwater ecosystems. The goal of this study was to determine the aquatic macrophyte Eichhornia azurea (Sw.) Kunth decomposition rate, the time relation with the limnological parameters, and whether this relationship is a result of decomposition processes. To that end, we collected water and leaves of E. azurea in Surf Leopoldo, PR. The experiment consisted of two treatments: 25 containers with 450mL of water and 0.8g of biomass dry weight were used with or without the addition of macrophytes. Samples were collected in triplicate at times 0, 3h, 6h, 12h, 24h, 72h, 120h, 168h and 240h. When the container was removed, the plant material was dried in an oven. After 48h, the material was measured to obtain the final dry weight. Analyses of pH, conductivity, dissolved oxygen, total phosphorus N-ammonia (NH4), soluble reactive phosphorus (PO4) and dissolved organic carbon were performed, and the decomposition rate was calculated. The results showed significant temporal variation of limnological parameters in the study. Additionally, dissolved oxygen, conductivity, dissolved organic carbon and total phosphorus were correlated with the dry weight of the biomass, suggesting that E. azurea decomposition significantly interferes with the dynamics of these variables.

Effects of different forms of nitrogen on rhizosphere microbial community structure of Eichhornia crassipes (Pontederiaceae)

AbstractRhizosphere microbial communities are important for phytoremediation, plant nutrition, health and metabolism. Many factors, including plant species, pH and nutritional factors influence rhizosphere microbiology. In this study, we analysed the effects of different forms of nitrogen on the structures of rhizosphere microbial communities of E. crassipes. Using a conventional culture method with special media, bacteria, actinobacteria and molds were cultured. We found that the numbers of bacteria were largely similar across the three culture conditions, while the numbers of actinobacteria and molds from the rhizosphere of E. crassipes cultured in NH4Cl solution were two orders of magnitude higher than those from the rhizospheres of plants cultured in distilled water and KNO3 solution. Using a culture-independent method of polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rDNA, we found that the form of nitrogen could influence the components of the rhizosphere microbial community. Pseudoxanthomonas, Enterobacter and Citrobacter were present in all of the samples cultured under the three different experimental conditions. The genus Reyranella was found only in samples cultured in KNO3 solution; Acinetobacter and Streptomyces were unique to samples cultured in NH4Cl solution, and Pseudomonas, Pseudacidovorax and Methylosinus were found only in samples cultured in distilled water. Pseudoxanthomonas and Acidovorax were the dominant genera in the rhizosphere microbial community of E. crassipes cultured in KNO3 solution, while Novosphingobium was the dominant genus in the sample cultured in a nitrogen-deficient medium. Our results provide a theoretical foundation for using E. crassipes as a phytoremediation plant and controlling the widespread distribution of E. crassipes around the world using principles of nutrient metabolism.

ResumenComunidades microbianas de la rizósfera son importantes para la fitorremediación, nutrición vegetal, salud y metabolismo. Muchos factores, incluyendo la especie de planta, el pH y los factores nutricionales influyen en la microbiología de la rizósfera. En este estudio, se analizaron los efectos de las diferentes formas del nitrógeno en la estructura de las comunidades microbianas de la rizósfera de E. crassipes. Mediante métodos de cultivo convencional con medios especiales se cultivaron: bacterias, actinobacterias y mohos. Se encontró que el número de bacterias era en gran parte similar a través de las tres condiciones de cultivo, mientras que el número de actinobacterias y mohos de la rizósfera de E. crassipes cultivadas en solución de NH4Cl era dos órdenes de magnitud superior a los de las rizósferas de plantas cultivadas en agua destilada y solución de KNO3. Utilizando un método de cultivo independiente de electroforesis en gel con gradiente de desnaturalización (PCR-DGGE) del ADNr 16S, se encontró que la forma de nitrógeno podría influir en los componentes de la comunidad microbiana de la rizósfera. Pseudoxanthomonas, Enterobacter y Citrobacter estaban presentes en todas las muestras cultivadas en las tres condiciones experimentales. El género Reyranella se encontró sólo en muestras cultivadas en solución de KNO3; Acinetobacter y Streptomyces eran las únicas muestras cultivadas en solución de NH4Cl, y Pseudomonas, Pseudacidovorax y Methylosinus se encontraron sólo en muestras cultivadas en agua destilada. Pseudoxanthomonas y Acidovorax eran los géneros dominantes en la comunidad microbiana de la rizósfera de E. crassipes cultivadas en solución de KNO3, mientras que Novos phingobium fue el género dominante en la muestra cultivada en un medio deficiente de nitrógeno. Nuestros resultados proporcionan una base teórica para el uso de E. crassipes como planta fitorremediadora y para controlar la distribución generalizada de E. crassipes en todo el mundo a través de los principios del metabolismo de nutrientes.

Effects of different forms of nitrogen on rhizosphere microbial community structure of Eichhornia crassipes (Pontederiaceae).

Rhizosphere microbial communities are important for phytoremediation, plant nutrition, health and metabolism. Many factors, including plant species, pH and nutritional factors influence rhizosphere micro- biology. In this study, we analysed the effects of different forms of nitrogen on the structures of rhizosphere microbial communities of E. crassipes. Using a conventional culture method with special media, bacteria, acti- nobacteria and molds were cultured. We found that the numbers of bacteria were largely similar across the three culture conditions, while the numbers of actinobacteria and molds from the rhizosphere of E. crassipes cultured in NH4Cl solution were two orders of magnitude higher than those from the rhizospheres of plants cultured in distilled water and KNO3 solution. Using a culture-independent method of polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rDNA, we found that the form of nitrogen could influence the components of the rhizosphere microbial community. Pseudoxanthomonas, Enterobacter and Citrobacter were present in all of the samples cultured under the three different experimental conditions. The genus Reyranella was found only in samples cultured in KNO(3) solution; Acinetobacter and Streptomyces were unique to samples cultured in NH(4)Cl solution, and Pseudomonas, Pseudacidovorax and Methylosinus were found only in samples cultured in distilled water. Pseudoxanthomonas and Acidovorax were the dominant genera in the rhizosphere microbial community of E. crassipes cultured in KNO(3) solution, while Novosphingobium was the dominant genus in the sample cultured in a nitrogen-deficient medium. Our results provide a theoretical foundation for using E. crassipes as a phytoremediation plant and controlling the widespread distribution of E. crassipes around the world using principles of nutrient metabolism.

Revealing crosstalk of plant and fungi in the symbiotic roots of sewage-cleaning Eichhornia crassipes using direct de novo metatranscriptomic analysis.

Cultivation and environmental changes can induce development of novel phenotypes in plants. For example, the root morphology of cultivated purple root Eichhornia crassipes differs remarkably from normal Eichhornia crassipes and also shows an enhanced ability to absorb heavy metal from groundwater. However, the changes in gene expression associated with these processes are unknown because of the lack of information on its large and unsequenced genome and its complex plant-rhizosphere symbiotic system. To investigate these gene expression changes, we applied a new strategy, direct de novo metatranscriptome analysis. Using this approach, we assembled the metatranscriptome of the entire rhizosphere and identified species-specific differentially expressed genes (DEGs) via hyper-accurate algorithms, showing a polarized plant/fungus distribution: the plant genes were responsible for morphological changes to the root system, offering a greater volume and surface area that hosts more fungi; while genes associated with heavy metal response in the fungus Fusarium were upregulated more than 3600-fold. These results suggested a distinct and synergistic functional response by the plant and fungal transcriptomes, indicating significant plant/fungal crosstalk during environmental changes. This study demonstrates that the metatranscriptomic approach adopted here offers a cost-efficient strategy to study symbiosis systems without the need for a priori genomic knowledge.

Removal of Chlorpyrifos by Water Hyacinth (Eichhornia crassipes) and the Role of a Plant-Associated Bacterium.

The objective of this research was to study the efficiency of water hyacinth (Eichhornia crassipes) and the role of any plant-associated bacteria in removing chlorpyrifos from water. The relative growth rate (RGR) of E. crassipes in the presence of 0.1 mg/L chlorpyrifos was not significantly different from that in its absence and only slightly decreased at concentrations of 0.5 and 1.0 mg/L by ∼1.1- and ∼1.2-fold, respectively, with an observed dry weight based RGRDW for E. crassipes of 0.036-0.041 mg/g/d. The removal rate constants of chlorpyrifos in the absence of plants were low at 3.52, 2.29 and 1.84 h(-1) for concentrations of 0.1, 0.5 and 1.0 mg/L, respectively, but were some 3.89- to 4.87-fold higher in the presence of E. crassipes. Chlorpyrifos removal was markedly facilitated by the presence of a root-associated bacterium, preliminarily identified as Acinetobacter sp. strain WHA. The interaction of E. crassipes and Acinetobacter sp. strain WHA provide an efficient and ecological alternative to accelerate the removal and degradation of chlorpyrifos pollution from aquatic systems including wastewater.

Polluting macrophytes Colombian lake Fúquene used as substrate by edible fungus Pleurotus ostreatus.

Invasive aquatic plants from Lake Fúquene (Cundinamarca, Colombia), water hyacinth (Eichhornia crassipes C. Mart.) and Brazilian elodea (Egeria densa Planch.) have been removed mechanically from the lake and can be used for edible mushrooms production. The growth of the oyster mushroom (Pleurotus ostreatus) on these aquatic macrophytes was investigated in order to evaluate the possible use of fruiting bodies and spent biomass in food production for human and animal nutrition, respectively. Treatments included: water hyacinth, Brazilian elodea, sawdust, rice hulls and their combinations, inoculated with P. ostreatus at 3%. Water hyacinth mixed with sawdust stimulated significantly fruiting bodies production (P = 3.3 × 10(-7)) with 71% biological efficacy, followed by water hyacinth with rice husk (55%) and elodea with rice husk (48%), all of these have protein contents between 26 and 47%. Loss of lignin (0.9-21.6%), cellulose (3.7-58.3%) and hemicellulose (1.9-53.8%) and increment in vitro digestibility (16.7-139.3%) and reducing sugars (73.4-838.4%) were observed in most treatments. Treatments spent biomass presented Relative Forage Values (RFV) from 46.1 to 232.4%. The results demonstrated the fungus degrading ability and its potential use in aquatic macrophytes conversion biomass into digestible ruminant feed as added value to the fruiting bodies production for human nutrition.

Cyanobacteria enhance methylmercury production: a hypothesis tested in the periphyton of two lakes in the Pantanal floodplain, Brazil.

The toxic potential of mercury (Hg) in aquatic systems is due to the presence and production of methylmercury (MeHg). Recent studies in tropical floodplain environments showed that periphyton associated with the roots of aquatic macrophytes produce MeHg. Periphyton communities are the first link in the food chain and one of the main MeHg sources in aquatic environments. The aim of this work was to test the hypotheses that the algal community structure affects potential methylation, and ecologically distinct communities with different algal and bacterial densities directly affect the formation of MeHg in the roots of macrophytes. To evaluate these, net MeHg production in the roots of Eichhornia crassipes in relation to the taxonomic structure of associated periphytic algae was evaluated. Macrophyte root samples were collected in the dry and flood season from two floodplain lakes in the Pantanal (Brazil). These lakes have different ecological conditions as a function of their lateral hydrological connectivity with the Paraguay River that is different during times of drought. Results indicated that MeHg production was higher in the flood season than in the dry season. MeHg production rates were higher in the disconnected lake in comparison to the connected lake during the dry season. MeHg production exhibited a strong positive co-variation with cyanobacteria abundance (R(2)=0.78; p<0.0001 in dry; R(2)=0.40; p=0.029 in flood) and with total algal biomass (R(2)=0.86; p<0.0001), and a negative co-variation with Zygnemaphyceae (R(2)=0.50; p=0.0018) in the lake community in dry season. This indicates that ecological conditions that favour the establishment and development of cyanobacteria are associated with higher rates of methylation in aquatic systems. This suggests that cyanobacteria could be a proxy for sites of MeHg production in some natural aquatic environments.

Fate of NO and NH in the treatment of eutrophic water using the floating macrophyte.

Use of the floating aquatic macrophyte, , to improve eutrophic water quality is practiced on a large scale in China. Limited information is available on the relative importance of the biological NO or NH removal process during the treatment of eutrophic water using . To investigate the key process responsible for the removal of NO and NH, N-NO (9.98 atom % [at.%] N) or N-NH (10.08 at.% N) was added to obtain eutrophic water with or without the cultivation of . In the unplanted water, considerable proportions of the added N-NO (27.13 ± 4.87%) or N-NH (42.08 ± 7.22%) were assimilated by the developed algae. The growth of controlled algae development in the planted water. Furthermore, the cultivation of stimulated gaseous loss of N by microbial denitrification (8.61 ± 1.70% NO-N loss from N-NO-labeled water). Apart from N loss by denitrification, considerable proportions of the added N-NO (62.01 ± 6.93%) or N-NH (76.76 ± 6.21%) were assimilated into the macrophyte N pools. The fine root detritus of contained a proportion of N (4.37 ± 1.39% in NO-labeled water, 2.03 ± 0.52% in NH-labeled water) that will be returned to the water after decomposition. In addition to N loss via NO emission, an unaccounted proportion of N could be mainly due to gaseous loss as N by denitrification (25.00% in N-NO-labeled water with Eichhornia crassipes)

Eichhornia crassipes capability to remove naphthalene from wastewater in the absence of bacteria.

The aim of the current study was to investigate the potential of an aquatic plant, the water hyacinth (Eichhornia crassipes) devoid rhizospheric bacteria, to reduce naphthalene (a polyaromatic hydrocarbon) present in wastewater and wetlands. The capability of sterile water hyacinth plants to remove naphthalene from water and wastewater was studied in batch systems. Water hyacinths enhance the removal of pollutants through their consumption as nutrients and also through microbial activity of their rhizospheric bacteria. Experimental kinetics of naphthalene removal by water hyacinth coupled with natural rhizospheric bacteria was 100% after 9 d. Plants, decoupled of rhizospheric bacteria, reduced naphthalene concentration up to 45% during 7 d. Additionally, naphthalene uptake by water hyacinth revealed a biphasic behavior: a rapid first phase completed after 2.5 h, and a second, considerably slower rate, phase (2.5-225 h). In conclusion, water hyacinth devoid rhizospheric bacteria reduced significantly naphthalene concentration in water, revealing a considerable plant contribution in the biodegradation process of this pollutant.

Mercury methylation and hydrogen sulfide production among unexpected strains isolated from periphyton of two macrophytes of the Amazon.

The periphyton of macrophytes had previously been identified as important spots for mercury methylation in the Amazon basin, but the microorganisms that facilitate methylation in such compartment are still to be identified. Here, bacteria were isolated from periphyton associated with Eichhornia crassipes and Polygonum densiflorum in Widdel and Pfennig medium and tested for mercury methylation with a stable isotope tracer technique using (198)HgCl, hydrogen sulfide production and molybdate inhibition. Three Pleomorphomona spp., one unidentified Deltaproteobacteria, two Klebsiella spp., and one Tolumonas sp. were isolated. All except Tolumonas sp. were able to methylate mercury (up to 5% of the (198)HgCl added) and produce up to 4 mM of H(2)S, while the Deltaproteobacteria was also able to demethylate methylmercury. Although these bacteria may not be as strong mercury methylators as sulfate-reducing bacteria, they have the potential to contribute to methylmercury accumulation in the system.

Optimization of pretreatment and saccharification for the production of bioethanol from water hyacinth by Saccharomyces cerevisiae.

Alkaline-oxidative (A/O) pretreatment and enzymatic saccharification were optimized for bioethanol fermentation from water hyacinth by Saccharomyces cerevisiae. Water hyacinth was subjected to A/O pretreatment at various NaOH and H(2)O(2) concentrations and reaction temperatures for the optimization of bioethanol fermentation by S. cerevisiae. The most effective condition for A/O pretreatment was 7% (w/v) NaOH at 100 °C and 2% (w/v) H(2)O(2). The carbohydrate content was analyzed after reaction at various enzyme concentrations and enzyme ratios using Celluclast 1.5 L and Viscozyme L to determine the effective conditions for enzymatic saccharification. After ethanol fermentation using S. cerevisiae KCTC 7928, the concentration of glucose, ethanol and glycerol was analyzed by HPLC using a RI detector. The yield of ethanol in batch fermentation was 0.35 g ethanol/g biomass. Continuous fermentation was carried out at a dilution rate of 0.11 (per h) and the ethanol productivity was 0.77 [g/(l h)].

Mercury methylation and the microbial consortium in periphyton of tropical macrophytes: effect of different inhibitors.

Macrophyte-associated periphyton is known as a site of Hg accumulation and methylation in tropical environments. Sulfate-reducing bacteria (SRB) is found in periphyton and its role in Hg methylation is acknowledged. However, the contribution of other microorganisms to this process is largely unknown. We tested the effect of inhibitors for different microorganisms on methylmercury (MMHg) formation on distinct macrophyte species from lakes of the Bolivian Amazon basin and in Brazil. We also tested the effect of inhibitors on bacterial secondary activity at two lakes in Brazil. Samples were incubated on-site with (203)Hg and Me(203)Hg was extracted and measured by liquid scintillation. MMHg formation on macrophytes varied among species ranging from 0.2% to 36%. Treatments with specific inhibitors resulted in reduction of MMHg production on most sites and inhibitors. The most successful treatment was the co-inhibition of SRB and methanogens. The inhibitions of algae and fungi activity showed fewer effects on methylation rates at all sites analyzed. Bacterial secondary activity was slightly affected by algae and fungi inhibition, and largely influenced by prokaryotic, SRB and methanogens inhibition. The data suggest that MMHg formation may not be directly performed by all microorganisms in periphyton but depends on complex interactions among them.

Molecular identification of two strains of Cercospora rodmanii isolated from water hyacinth present in Yuriria lagoon, Guanajuato, Mexico and identification of new hosts for several other strains.

Water hyacinth is a beautiful monocotyledon plant that has been dispersed all over the world by humans. The plant has been present in Mexico since 1907, and many water bodies have become infested with it since then. In 2001, we initiated a survey in Yuriria lagoon in southern Guanajuato state to isolate fungi able to biocontrol the plant. We isolated 25 morphologically distinct fungal cultures, of which two were identified as members of the genus Cercospora. Cercospora species are among the most prevalent and destructive of plant pathogens and can be found on leaves, pedicels, stems, fruits, and bracts. Only two species of Cercospora, Cercospora piaropi, and Cercospora rodmanii, have been described on water hyacinth; however, the classification of these species has been controversial. Several molecular approaches have been used for Cercospora identification, and some candidate genes have been identified for use in Cercospora species determination. Although the nrRNA genes alone do not show sufficient resolution for species determination, histone H3, translation elongation factor1-α, ß-tubulin, actin, and calmodulin have been shown in previous studies to have an adequate number of nucleotide changes to allow species identification. In the present study, we used partial sequences of the histone H3, actin, and calmodulin genes to identify our two isolates as C. rodmanii. Our two strains are not specific to water hyacinth, as they are also pathogenic to beet and sugar beet. Similar host ranges were found for C. rodmanii strains isolated from Tabasco in México, Zambia, and Brazil, however, the specificity for water hyacinth persists in Cercospora piaropi Tharp and C. rodmanii Conway, the latter being the most pathogenic.

Importance of sulfate reducing bacteria in mercury methylation and demethylation in periphyton from Bolivian Amazon region.

Sulfate reducing bacteria (SRB) are important mercury methylators in sediments, but information on mercury methylators in other compartments is ambiguous. To investigate SRB involvement in methylation in Amazonian periphyton, the relationship between Hg methylation potential and SRB (Desulfobacteraceae, Desulfobulbaceae and Desulfovibrionaceae) abundance in Eichhornia crassipes and Polygonum densiflorum root associated periphyton was examined. Periphyton subsamples of each macrophyte were amended with electron donors (lactate, acetate and propionate) or inhibitors (molybdate) of sulfate reduction to create differences in SRB subgroup abundance, which was measured by quantitative real-time PCR with primers specific for the 16S rRNA gene. Mercury methylation and demethylation potentials were determined by a stable isotope tracer technique using 200HgCl and CH3(202)HgCl, respectively. Relative abundance of Desulfobacteraceae (<0.01-12.5%) and Desulfovibrionaceae (0.01-6.8%) were both highly variable among samples and subsamples, but a significant linear relationship (p<0.05) was found between Desulfobacteraceae abundance and net methylmercury formation among treatments of the same macrophyte periphyton and among all P. densiflorum samples, suggesting that Desulfobacteraceae bacteria are the most important mercury methylators among SRB families. Yet, molybdate only partially inhibited mercury methylation potentials, suggesting the involvement of other microorganisms as well. The response of net methylmercury production to the different electron donors and molybdate was highly variable (3-1104 pg g(-1) in 12 h) among samples, as was the net formation in control samples (17-164 pg g(-1) in 12 h). This demonstrates the importance of community variability and complexity of microbial interactions for the overall methylmercury production in periphyton and their response to external stimulus.

Does water hyacinth on East African lakes promote cholera outbreaks?

Cholera outbreaks continue to occur regularly in Africa. Cholera has been associated with proximity to lakes in East Africa, and Vibrio cholerae has been found experimentally to concentrate on the floating aquatic plant, water hyacinth, which is periodically widespread in East African lakes since the late 1980s. From 1994 to 2008, Nyanza Province, which is the Kenyan province bordering Lake Victoria, accounted for a larger proportion of cholera cases than expected by its population size (38.7% of cholera cases versus 15.3% of national population). Yearly water-hyacinth coverage on the Kenyan section of Lake Victoria was positively associated with the number of cholera cases reported in Nyanza Province (r = 0.83; P = 0.0010). Water hyacinth on freshwater lakes might play a role in initiating cholera outbreaks and causing sporadic disease in East Africa.

Isolation, characterization, and production of red pigment from Cercospora piaropi a biocontrol agent for waterhyacinth.

A red pigment produced by a Mexican isolate of Cercospora piaropi (waterhyacinth pathogen) has been isolated and identified as cercosporin. The kinetic of cercosporin production in culture media during dark/light regimes was evaluated. When C. piaropi was cultivated in continuous light and potato dextrose broth culture, a maximum of cercosporin production was observed (72.59 mg/l). Despite other reports, C piaropi Mexican isolate produce cercosporin in dark conditions (25.70 mg/l). The results suggest that production of cercosporin in C. piaropi-waterhyacinth pathogenesis is an important factor to take into account in biocontrol strategies.

Enterotoxigenic Escherichia coli in sewage-impacted waters and aquatic weeds: quantitative PCR for culture-independent enumeration.

AIM: To develop quantitative PCR for culture-independent enumeration of enterotoxigenic Escherichia coli (ETEC) in sewage-impacted waters and aquatic weeds. METHODS AND RESULTS: Two fluorescent probes (TaqMan and FRET) based on two different real-time PCR chemistries were designed in highly conserved region of LT1 gene encoding heat labile enterotoxin. Both the assays could detect 2 CFU ml(-1) from serially diluted (two-fold and ten-fold) culture of reference strain (E. coli MTCC 723). FRET performed better in terms of CT value and PCR efficiency than TaqMan. The presence of 10(6) CFU ml(-1) of nonpathogenic E. coli reduced the detection limit two-fold with both the probes. However, the performance for two chemistries in various environmental samples was significantly (student's t-test, P<0.05) different. CONCLUSION: It could be inferred from this study that real-time PCR chemistries (TaqMan and FRET) could detect very few copies of target DNA in pure cultures, but may give varied response in the presence of nonspecific DNA and natural inhibitors present in environmental sample matrices. SIGNIFICANCE AND IMPACT OF THE STUDY: The assays can be used for pre-emptive monitoring of aquatic weeds (a potential nonpoint source), surface and potable waters to prevent waterborne outbreaks caused by ETEC.