Physiological characterization of Mycobacterium sp. strain 1B isolated from a bacterial culture able to degrade high-molecular-weight polycyclic aromatic hydrocarbons

AIM: The aim of this study was to further characterize a bacterial culture (VUN 10,010) capable of benzo[a]pyrene cometabolism. METHODS AND RESULTS: The bacterial culture, previously characterized as a pure culture of Stenotrophomonas maltophilia (VUN 10,010), was found to also contain another bacterial species (Mycobacterium sp. strain 1B), capable of degrading a similar range of PAH substrates. Analysis of its 16S rRNA gene sequence and growth characteristics revealed the strain to be a fast-growing Mycobacterium sp., closely related to other previously isolated PAH and xenobiotic-degrading mycobacterial strains. Comparison of the PAH-degrading characteristics of Mycobacterium sp. strain 1B with those of S. maltophilia indicated some similarities (ability to degrade phenanthrene and pyrene), but some differences were also noted (S. maltophilia able to degrade fluorene, but not fluoranthene, whereas Mycobacterium sp. strain 1B can degrade fluoranthene, but not fluorene). Unlike the S. maltophilia culture, there was no evidence of benzo[a]pyrene degradation by Mycobacterium sp. strain 1B, even in the presence of other PAHs (ie pyrene) as co-metabolic substrates. Growth of Mycobacterium sp. strain 1B on other organic carbon sources was also limited compared with the S. maltophilia culture. CONCLUSIONS: This study isolated a Mycobacterium strain from a bacterial culture capable of benzo[a]pyrene cometabolism. The Mycobacterium strain displays different PAH-degrading characteristics to those described previously for the PAH-degrading bacterial culture. It is unclear what role the two bacterial strains play in benzo[a]pyrene cometabolism, as the Mycobacterium strain does not appear to have endogenous benzo[a]pyrene degrading ability. SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes the isolation and characterization of a novel PAH-degrading Mycobacterium strain from a PAH-degrading culture. Further studies utilizing this strain alone, and in combination with other members of the consortium, will provide insight into the diverse roles different bacteria may play in PAH degradation in mixed cultures and in the environment.     

Isolation and characterization of a bacterial strain of the genus Ochrobactrum with methyl parathion mineralizing activity

AIMS: To isolate and characterize a methyl parathion (MP)-mineralizing bacterium, and to elucidate the degradative pathway of MP and localize the responsible degrading genes. METHODS AND RESULTS: A bacterial strain, designated B2, capable of mineralizing MP was isolated from the MP-polluted soil. Analysis of the 16S rRNA gene sequence and phenotypic analysis suggested that strain B2 had a close relationship with Ochrobactrum anthropi. B2 could totally degrade MP and four metabolites [p-nitrophenol (PNP), 4-nitrocatechol (4-NC), 1,2,4-benzenetriol (BT) and hydroquinone (HQ)] were identified by HPLC and gas chromatography-mass spectrometry analyses. Plasmid curing of strain B2 resulted in the loss of ability of B2 to degrade PNP, but not the ability to hydrolyse MP. CONCLUSIONS: Ochrobactrum sp. B2 can mineralize MP rapidly via PNP, 4-NC, BT and HQ pathway. B2 harbours a plasmid encoding the ability to degrade PNP, while MP-hydrolysing activity is encoded on the bacterial chromosome. SIGNIFICANCE AND IMPACT OF THE STUDY: This new bacterial strain (B2) capable of mineralizing MP will be useful in a pure-culture remediation process of organophosphate pesticides and their metabolites such as nitroaromatics.     

Potential application of Pistia stratiotes for the phytoremediation of mesotrione and its degradation products from water

Abstract

The aim of the present work is to estimate remediation potential of Pistia stratiotes, its ability to uptake mesotrione (MES) - one of the most frequently used herbicides, and its main degradation products: 2-amino-4-methylsulfonyl benzoic acid (AMBA) and 4-methylsulfonyl-2-nitrobenzoic acid (MNBA). This research focuses on model experiments performed under laboratory conditions. The results show that Pistia stratiotes can uptake up to 75% of degradation products from 1?L of surface water samples polluted with 0.4?µg/L of each analyte during 7?days without significant phytotoxic effect. Under the same experimental conditions, the effectiveness of mesotrione sorption is in the range of 42–58%. The phytotoxicity of this compound is higher in comparison to its degradation products (decrease of chlorophyll concentration in plant tissues exposed to MES 27–32% vs 4–13% in case of exposition to AMBA and MNBA). The adequate nutrition of the plants is crucial to their well-being and thus the sorption of pollutants.     

A novel pathway for the biodegradation of gamma-hexachlorocyclohexane by a Xanthomonas sp. strain ICH12

AIM: To isolate gamma-hexachlorocyclohexane (HCH)-degrading bacteria from contaminated soil and characterize the metabolites formed and the genes involved in the degradation pathway. METHODS AND RESULTS: A bacterial strain Xanthomonas sp. ICH12, capable of biodegrading gamma- HCH was isolated from HCH-contaminated soil. DNA-colony hybridization method was employed to detect bacterial populations containing specific gene sequences of the gamma-HCH degradation pathway. linA (dehydrodehalogenase), linB (hydrolytic dehalogenase) and linC (dehydrogenase) from a Sphingomonas paucimobilis UT26, reportedly possessing gamma-HCH degradation activity, were used as gene probes against isolated colonies. The isolate was found to grow and utilize gamma-HCH as the sole carbon and energy source. The 16S ribosomal RNA gene sequence of the isolate resulted in its identification as a Xanthomonas species, and we designated it as strain ICH12. During the degradation of gamma-HCH by ICH12, formation of two intermediates, gamma-2,3,4,5,6-pentachlorocyclohexene (gamma-PCCH), and 2,5-dichlorobenzoquinone (2,5-DCBQ), were identified by gas chromatography-mass spectrometric (GC-MS) analysis. While gamma-PCCH was reported previously, 2,5-dichlorohydroquinone was a novel metabolite from HCH degradation. CONCLUSIONS: A Xanthomonas sp. for gamma-HCH degradation from a contaminated soil was isolated. gamma-HCH was utilized as sole source of carbon and energy, and the degradation proceeds by successive dechlorination. Two degradation products gamma-PCCH and 2,5-DCBQ were characterized, and the latter metabolite was not known in contrasts with the previous studies. The present work, for the first time, demonstrates the potential of a Xanthomonas species to degrade a recalcitrant and widespread pollutant like gamma-HCH. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the isolation and characterization of a novel HCH-degrading bacterium. Further results provide an insight into the novel degradation pathway which may exist in diverse HCH-degrading bacteria in contaminated soils leading to bioremediation of gamma-HCH.     

Isolation and characterization of an atrazine-degrading bacterium from industrial wastewater in China

AIMS: To isolate and characterize atrazine-degrading bacteria in order to identify suitable candidates for potential use in bioremediation of atrazine contamination. METHODS AND RESULTS: A high efficiency atrazine-degrading bacterium, strain AD1, which was capable of utilizing atrazine as a sole nitrogen source for growth, was isolated from industrial wastewater. 16S rDNA sequencing identified AD1 as an Arthrobacter sp. The atrazine chlorohydrolase gene (atzA) isolated from strain AD1 differed from that found in the Pseudomonas sp. ADP by only one nucleotide. However, it was found located on the bacterial chromosome rather than on plasmids as previously reported for other bacteria. CONCLUSIONS: Atrazine chlorohydrolase gene, atzA, either encoded by chromosome or plasmid, is highly conserved. SIGNIFICANCE AND IMPACT OF THE STUDY: Comparison analysis of atrazine degradation gene structure and arrangement in this and other bacteria provides insight into our understanding of the ecology and evolution of atrazine-degrading bacteria.     

Enantioselective synthesis of S-equol from dihydrodaidzein by a newly isolated anaerobic human intestinal bacterium

A newly isolated rod-shaped, gram-negative anaerobic bacterium from human feces, named Julong 732, was found to be capable of metabolizing the isoflavone dihydrodaidzein to S-equol under anaerobic conditions. The metabolite, equol, was identified by using electron impact ionization mass spectrometry, (1)H and (13)C nuclear magnetic resonance spectroscopy, and UV spectral analyses. However, strain Julong 732 was not able to produce equol from daidzein, and tetrahydrodaidzein and dehydroequol, which are most likely intermediates in the anaerobic metabolism of dihydrodaidzein, were not detected in bacterial culture medium containing dihydrodaidzein. Chiral stationary-phase high-performance liquid chromatography eluted only one metabolite, S-equol, which was produced from a bacterial culture containing a racemic mixture of dihydrodaidzein. Strain Julong 732 did not show racemase activity to transform R-equol to S-equol and vice versa. Its full 16S rRNA gene sequence (1,429 bp) had 92.8% similarity to that of Eggerthella hongkongenis HKU10. This is the first report of a single bacterium capable of converting a racemic mixture of dihydrodaidzein to enantiomeric pure S-equol.     

Isolation and identification of a novel valienamine-producing bacterium

AIMS: To isolate, identify and assess valienamine production by a soil bacterial isolate from a wheat field in Hangzhou, China. METHODS AND RESULTS: A validamycin A-degrading bacterial strain, numbered ZJB-041, was isolated and identified as Stenotrophomonas maltophilia, based on morphology, physiological tests, ATB system (ID32 GN), and 16S rDNA analysis. The strain was capable of producing valienamine by decomposing validamycin A. After fermentation in shaking flasks at 30 degrees C for 7 days, 96.0% of 34.49 mmol l(-1) of validamycin A was degraded and 2.65 mmol l(-1) of valienamine was obtained. The resting cells of this strain also produced valienamine by degrading validamycin A. After 72 h of incubation in 0.2 mol l(-1) of phosphate buffer (pH 7.5), 90.2% of 17.16 mmol l(-1) of validamycin A was degraded, and 1.77 mmol l(-1) of valienamine was obtained. CONCLUSIONS: Our data suggested that S. maltophilia ZJB-041, a bacterial isolate, has the potential for validamycin A degradation and valienamine production. SIGNIFICANCE AND IMPACT OF THE STUDY: The validamycin A-degrading bacterium could potentially be utilized in the disposal of validamycin residues and in the production of valienamine.     

Isolation and characterisation of an equol-producing mixed microbial culture from a human faecal sample and its activity under gastrointestinal conditions

Only about one third of humans possess a microbiota capable of transforming the dietary isoflavone daidzein into equol. Little is known about the dietary and physiological factors determining this ecological feature. In this study, the in vitro metabolism of daidzein by faecal samples from four human individuals was investigated. One culture produced the metabolites dihydrodaidzein and O-desmethylangolensin, another produced dihydrodaidzein and equol. From the latter, a stable and transferable mixed culture transforming daidzein into equol was obtained. Molecular fingerprinting analysis (denaturing gradient gel electrophoresis) showed the presence of four bacterial species of which only the first three strains could be brought into pure culture. These strains were identified as Lactobacillus mucosae EPI2, Enterococcus faecium EPI1 and Finegoldia magna EPI3, and did not produce equol in pure culture. The fourth species was tentatively identified as Veillonella sp strain EP. It was found that hydrogen gas in particular, but also butyrate and propionate, which are all colonic fermentation products from poorly digestible carbohydrates, stimulated equol production by the mixed culture. However, when fructo-oligosaccharides were added, equol production was inhibited. Furthermore, the equol-producing capacity of the isolated culture was maintained upon its addition to a faecal culture originating from a non-equol-producing individual.     

Biodegradation of polyethylene by the thermophilic bacterium Brevibacillus borstelensis

AIM: To select a polyethylene-degrading micro-organism and to study the factors affecting its biodegrading activity. METHODS AND RESULTS: A thermophilic bacterium Brevibaccillus borstelensis strain 707 (isolated from soil) utilized branched low-density polyethylene as the sole carbon source and degraded it. Incubation of polyethylene with B. borstelensis (30 days, 50 degrees C) reduced its gravimetric and molecular weights by 11 and 30% respectively. Brevibaccillus borstelensis also degraded polyethylene in the presence of mannitol. Biodegradation of u.v. photo-oxidized polyethylene increased with increasing irradiation time. Fourier Transform Infra-Red (FTIR) analysis of photo-oxidized polyethylene revealed a reduction in carbonyl groups after incubation with the bacteria. CONCLUSIONS: This study demonstrates that polyethylene--considered to be inert--can be biodegraded if the right microbial strain is isolated. Enrichment culture methods were effective for isolating a thermophilic bacterium capable of utilizing polyethylene as the sole carbon and energy source. Maximal biodegradation was obtained in combination with photo-oxidation, which showed that carbonyl residues formed by photo-oxidation play a role in biodegradation. Brevibaccillus borstelensis also degraded the CH2 backbone of nonirradiated polyethylene. SIGNIFICANCE AND IMPACT OF THE STUDY: Biodegradation of polyethylene by a single bacterial strain contributes to our understanding of the process and the factors affecting polyethylene biodegradation.     

Cooperative biodegradation of geosmin by a consortium comprising three gram-negative bacteria isolated from the biofilm of a sand filter column

AIMS: To isolate and identify bacteria from a sand filter column capable of degrading the taste and odour compound, geosmin. In doing so, to investigate if these organisms degrade geosmin either individually or if an alternative mechanism is utilized. METHODS AND RESULTS: Geosmin-degrading bacteria from a biologically active sand filter column were enriched by their growth in a minimal medium supplemented with geosmin as the sole carbon source. By day 51, 21.7 mg l(-1) of geosmin had been degraded as determined by solid-phase microextraction gas chromatography/mass spectrometry, and was accompanied by a 2.12 log(10) increase in active bacterial numbers as measured using the BacLight(TM) bacterial viability kit and flow cytometric enumeration. During the onset of geosmin degradation, the predominance of three bacteria, most similar to previously cultured species of Sphingopyxis alaskensis, Novosphingobium stygiae and Pseudomonas veronii based on 16S rRNA gene sequences was detected by denaturing gradient gel electrophoresis. Subsequent isolation of these organisms revealed that degradation of geosmin, when present as either the sole carbon source (ranging from 40 ng l(-1) to 20 mg l(-1)) or when spiked into sterile reservoir water (37 and 131 ng l(-1)), occurred only when all three isolates were present. None of the isolates was shown to be capable of degrading geosmin either individually or in any combination of two. CONCLUSIONS: This study has reported, for the first time, the cooperative degradation of geosmin by a consortium comprising three gram-negative bacteria isolated from a biologically active sand filter column. SIGNIFICANCE AND IMPACT OF THE STUDY: These results are important for researchers currently employing molecular-based approaches to further understand the biodegradation of geosmin by bacteria, as such studies may be complicated by the discovery of geosmin degradation occurring by a consortium. This study also advances the knowledge surrounding the types of bacteria capable of degrading the taste and odour compound, as investigations to date regarding this are limited.     

Streptomyces sp. strain isolated from river water has high bisphenol A degradability

AIMS: We report the identification of the bisphenol A (BPA) biodegradability in Streptomyces sp. strain isolated from river water. METHODS AND RESULTS: The water samples spiked with BPA (1 mg l(-1)) and the culture solution of Streptomyces sp. strain were placed at 30 degrees C for 10 days and were analysed by high-performance liquid chromatography. A half-life for BPA degradation was between 3 and 4 days. The removal rate of BPA was >90% for 10 days. CONCLUSIONS: These results show that the Streptomyces sp. strain isolated from river water has high BPA degradability. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report of BPA degradation by Streptomyces sp. strain.     

Identification of the culturable and nonculturable bacterial population in ground water of a municipal water supply in Germany

AIMS: The identification of the culturable and nonculturable bacterial population in ground water of a municipal water supply in Mainz (Germany) during the year 2002. METHODS AND RESULTS: Total counts varied between 3.5 x 103 and 2.2 x 104 cells ml-1, viable counts were approximately between 8.1 x 102 and 3.3 x 103 cells ml-1. After cultivation on different nutrient media (R2A, DEV, PCA, Endo, Standard) <1% appeared to be culturable on the media used. After denaturating gradient gel electrophoreses, up to 24 different bacterial species were detected in the ground water. With the aid of 16S rDNA isolation, amplification and sequencing, the isolated organisms and clones could be identified. CONCLUSIONS: The isolated and cultured organisms mainly belonged to the Proteobacteria (alpha, beta and gamma), Flavobacteria or Actinobacteria. However, most of the noncultured micro-organisms were beta-Proteobacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study in which the identification of all culturable and nonculturable bacteria in a ground water has been attempted.     

Isolation and characterization of a lipolytic bacterium capable of growing in a low-water-content oil-water emulsion

A unique lipolytic bacterium was isolated in a selective growth system consisting of 99% triglycerides and a 1% water phase. The bacterium, termed Pseudomonas aeruginosa YS-7, was able to grow in an environment of low water content and could also survive amphipathic, osmotic, and matrical water stress in a triglyceride-rich culture. The isolated strain was identified as P. aeruginosa on the basis of standard physiological, biochemical, and serological assays. The strain is a gram-negative motile rod, aerobic, pigment forming, and capable of growing at 42 degrees C. It is highly tolerant of high concentrations of the cationic detergent cetyltrimethylammonium bromide and of the fatty acid salts derived from bacterial hydrolysis of the oil. Growth of the bacterium in a pure culture in a 99% triglyceride medium lasted until most of the water was evaporated or consumed. Growth was accompanied by triglyceride hydrolysis, which continued to occur even after growth saturation until the water was totally depleted. No loss of viability was observed when the culture was maintained under water-depleted conditions for an additional 40 h. A second cycle of bacterial growth and triglyceride hydrolysis was immediately initiated upon the addition of 1% (vol/vol) water to the culture. Lipase activity was stable regardless of changes in culture conditions. The isolated strain is uniquely resistant to severe water stress in a triglyceride-rich medium or under cold acetone precipitation compared with 12 other microbial strains, including bacteria and yeasts. Among these 12, only the lipolytic strains grew in the 99% triglyceride medium, but they reached a cell mass fourfold smaller than that of P. aeruginosa YS-7.(ABSTRACT TRUNCATED AT 250 WORDS)     

Major secondary metabolites produced by two commercial Trichoderma strains active against different phytopathogens

AIMS: Trichoderma harzianum strains T22 and T39 are two micro-organisms used as active agents in a variety of commercial biopesticides and biofertilizers and widely applied amongst field and greenhouse crops. The production, isolation, biological and chemical characterization of the main secondary metabolites produced by these strains are investigated. METHODS AND RESULTS: Of the three major compounds produced by strain T22, one is a new azaphilone that shows marked in vitro inhibition of Rhizoctonia solani, Pythium ultimum and Gaeumannomyces graminis var. tritici. In turn, filtrates from strain T39 were demonstrated to contain two compounds previously isolated from other T. harzianum strains and a new butenolide. The production of the isolated metabolites was also monitored by liquid chromatography/mass spectrometry during in vitro interaction with R. solani. CONCLUSIONS: This paper reports the isolation and characterization of the main secondary metabolites obtained from culture filtrates of two T. harzianum strains and their production during antagonistic interaction with the pathogen R. solani. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first work on secondary metabolites produced by the commercially applied strains T22 and T39. Our results provide a better understanding of the metabolism of these fungi, which are both widely used as biopesticides and/or biofertilizers in biocontrol.     

Isolation and characterization of a lipolytic bacterium capable of growing in a low-water-content oil-water emulsion.

A unique lipolytic bacterium was isolated in a selective growth system consisting of 99% triglycerides and a 1% water phase. The bacterium, termed Pseudomonas aeruginosa YS-7, was able to grow in an environment of low water content and could also survive amphipathic, osmotic, and matrical water stress in a triglyceride-rich culture. The isolated strain was identified as P. aeruginosa on the basis of standard physiological, biochemical, and serological assays. The strain is a gram-negative motile rod, aerobic, pigment forming, and capable of growing at 42 degrees C. It is highly tolerant of high concentrations of the cationic detergent cetyltrimethylammonium bromide and of the fatty acid salts derived from bacterial hydrolysis of the oil. Growth of the bacterium in a pure culture in a 99% triglyceride medium lasted until most of the water was evaporated or consumed. Growth was accompanied by triglyceride hydrolysis, which continued to occur even after growth saturation until the water was totally depleted. No loss of viability was observed when the culture was maintained under water-depleted conditions for an additional 40 h. A second cycle of bacterial growth and triglyceride hydrolysis was immediately initiated upon the addition of 1% (vol/vol) water to the culture. Lipase activity was stable regardless of changes in culture conditions. The isolated strain is uniquely resistant to severe water stress in a triglyceride-rich medium or under cold acetone precipitation compared with 12 other microbial strains, including bacteria and yeasts. Among these 12, only the lipolytic strains grew in the 99% triglyceride medium, but they reached a cell mass fourfold smaller than that of P. aeruginosa YS-7.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interactions between the unicellular red alga Rhodella reticulata (Rhodophyta) and contaminated bacteria

AIMS: To define the role of the bacterial strains LR1 and LR3 in the Rhodella cell destruction caused by Cytophaga sp.LR2. METHODS AND RESULTS: The bacteria were obtained from algal culture with destruction. They were isolated in pure culture and tested for biochemical activities using Polymicrotest. The ability of bacteria to degrade and utilize the algal polysaccharide was investigated. The bacteria were grown in a media containing Rhodella polysaccharide as a sole carbon source. The level of the reducing sugars in the culture media was determined. Scanning electron microscopy (SEM) was used to define the location of bacteria in extensively and intensively cultivated Rhodella reticulata previously infected by Cytophaga sp. LR2. CONCLUSIONS: The lysis of Rhodella reticulata cells is due to the joint action of the three bacterial strains with the former pathogen Cytophaga sp. LR2 playing the main role. The accumulation of the polysaccharide and the excreted metabolites of the strains LR1 and LR3 stimulated the development of Cytophaga sp. LR2. The adaptation of the strain to particular conditions of alga cultivation and the utilization of polysaccharide as a sole carbon source supported its stable growth in alga suspension and destruction of Rhodella cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The predominance of Cytophaga sp. LR2 over the two other contaminants and the lysis of Rhodella reticulata cells resulted from the ability of the bacterium to attach to the algal polysaccharide sheath. The formation of slime and extrusions facilitated the phenomenon of bacterial adhesion to the algal surface as well as the formation of colonial alga - bacterial spherules. The sedimentation of these aggregates decreased the ability of the algal strain to photosynthesize, led to the lysis of the cells and finally caused the death of Rhodella.     

Biological control agent of common scab disease by antagonistic strain Bacillus sp. sunhua

AIMS: To identify an antagonistic strain against Streptomyces scabiei and to characterize the antibiotic agent. The efficacy of the isolated strain in controlling common scab disease was also evaluated. METHODS AND RESULTS: A bacterial strain antagonistic against S. scabiei was isolated from the soil of a potato-cultivating area. This bacterium was identified as a Bacillus species by 16S rRNA gene sequence analysis and was designated Bacillus sp. sunhua. Antibiotics produced by this strain were proven to be stable within a broad pH range and at high temperatures. The culture broth was extracted with ethyl acetate, and then the crude extract was applied to HPLC. Two compounds were isolated and identified as iturin A and macrolactin A by 1H-NMR, 13C-NMR, HMBC, HMQC and mass spectrometer. The culture broth of Bacillus sp. sunhua had a suppressive effect on common scab disease in a pot assay, decreasing the infection rate from 75 to 35%. This strain also suppressed Fusarium oxysporum, the pathogen of potato dry rot disease. CONCLUSIONS: Bacillus sp. sunhua was shown to inhibit S. scabiei effectively. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report demonstrating that macrolactin A and iturin A inhibit S. scabiei. This study demonstrated the possibility of controlling potato scab disease using Bacillus sp. sunhua.     

Combined use of 16S ribosomal DNA and automated ribosomal intergenic spacer analysis to study the bacterial community in catfish ponds

AIMS: To apply culture-independent techniques to explore the bacterial community composition in catfish pond water. METHODS AND RESULTS: 16S rDNA libraries were constructed and sequenced from 15 pond water samples. Automated ribosomal intergenic spacer analysis (ARISA) was used to fingerprint each bacterial community. A broad diversity in bacterial species composition was found by 16S rDNA analysis. Alphaproteobacteria was the most represented class in all ponds, followed by Gammaproteobacteria and Gram-positive high G + C content bacteria. Uniqueness of bacterial communities from each individual pond was confirmed by ARISA. Catfish pathogens were detected sporadically. CONCLUSIONS: Bacterial communities in a catfish aquaculture setting can vary from pond to pond at one given point. No correlation could be made between bacteria composition and fish strain or between bacterial profile and the presence of catfish pathogens in a particular pond. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report showing the composition of bacterial communities in catfish ponds. Fish health specialists and catfish aquaculture managers should be aware of the wide differences in bacterial communities between ponds and include this variable in fish husbandry practices.     

rpoB-PCR amplified gene and temporal temperature gradient gel electrophoresis: a rapid tool to analyse bacterial strains representative of cold-smoked salmon microflora

AIM: To evaluate rpoB gene as a biomarker of microbial biodiversity associated to cold-smoked salmon by a novel nested-polymerase chain reaction/temporal temperature gradient gel electrophoresis (PCR/TTGE) technique applied on pure cultures of reference strains. METHODS AND RESULTS: DNA obtained from pure cultures of reference strains was used in a succession of a first PCR amplification of rpoB fragment with degenerated nonclamped primers and a nested-PCR with nondegenerated clamped primers. PCR products were then applied on a TTGE gel in order to analyse strains profile. High quantity of nested-PCR products were obtained for each tested strain and TTGE profiles showed a good separation between the different reference bacteria and an easy way to associate one band to one species. CONCLUSION: The nested-PCR/TTGE technique used in this study is a promising way of investigating bacterial community structure of cold-smoked salmon or other food matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its single copy state leading to single band profiles in TTGE, rpoB constitute a good potential molecular marker for further development of cold-smoked salmon biodiversity analysis.     

Isolation and characterization of a Mycobacterium strain that metabolizes the insecticide endosulfan

AIM: The aim of this study was to isolate and characterize a bacterium capable of metabolizing endosulfan. METHODS AND RESULTS: A endosulfan-degrading bacterium (strain ESD) was isolated from soil inoculum after repeated culture with the insecticide as the sole source of sulfur. Analysis of its 16S rRNA gene sequence, and morphological and physiological characteristics revealed it to be a new fast-growing Mycobacterium, closely related to other Mycobacterium species with xenobiotic-degrading capabilities. Degradation of endosulfan by strain ESD involved both oxidative and sulfur-separation reactions. Strain ESD did not degrade endosulfan when sulfite, sulphate or methionine were present in the medium along with the insecticide. Partial degradation occurred when the culture was grown, with endosulfan, in the presence of MOPS (3-(N-morpholino)propane sulphonic acid), DMSO (dimethyl sulfoxide), cysteine or sulphonane and complete degradation occurred in the presence of gutathione. When both beta-endosulfan and low levels of sulphate were provided as the only sources of sulfur, biphasic exponential growth was observed with endosulfan metabolism being restricted to the latter phase of exponential growth. CONCLUSIONS: This study isolated a Mycobacterium strain (strain ESD) capable of metabolizing endosulfan by both oxidative and sulfur-separation reactions. The endosulfan-degrading reactions are a result of the sulfur-starvation response of this bacterium. SIGNIFICANCE AND IMPACT OF THE STUDY: This describes the isolation of a Mycobacterium strain capable of degrading the insecticide endosulfan. This bacterium is a valuable source of enzymes for use in enzymatic bioremediation of endosulfan residues.