Genomic features of Pseudomonas sp. RGM2144 correlates with increased survival of rainbow trout infected by Flavobacterium psychrophilum.

This study evaluated the probiotic potential of the biofilm formed by the strain Pseudomonas sp. RGM2144 on rainbow trout survival. When challenged with the fish pathogen Flavobacterium psychrophilum, Pseudomonas sp. RGM2144 increased rainbow trout survival to 92.7 ± 1.2% (control: 35.3 ± 9.5%, p < .0001). The draft genome of Pseudomonas sp. RGM2144 is 6.8 Mbp long, with a completeness 100% and a contamination of 0.4%. The genome contains 6122 protein-coding genes of which 3564 (~60%) have known functions. The genome and phylogeny indicate that Pseudomonas sp. RGM2144 is a new species in the Pseudomonas genus, with few virulence factors, plasmids, and genes associated with antimicrobial resistance, suggesting a non-pathogenic bacterium with protective potential. In addition, the genome encodes for 11 secondary metabolite biosynthetic gene clusters that could be involved in the inhibition of F. psychrophilum. We suggest that Pseudomonas sp. RGM2144 may be applied as a probiotic in salmonid fish farming.

Piscirickettsia salmonis forms a biofilm on nylon surface using a CDC Biofilm Reactor.

Research into Piscirickettsia salmonis biofilms on materials commonly used in salmon farming is crucial for understanding its persistence and virulence. We used the CDC Biofilm Reactor to investigate P. salmonis (LF-89 and EM-90) biofilm formation on Nylon, Stainless steel (316L), Polycarbonate and High-Density Polyethylene (HDPE) surfaces. After 144 h of biofilm visualization by scanning confocal laser microscopy under batch growth conditions, Nylon coupons generated the greatest biofilm formation and coverage compared to Stainless steel (316L), Polycarbonate and HDPE. Additionally, P. salmonis biofilm formation on Nylon was significantly greater (p ≤ .01) than Stainless steel (316L), Polycarbonate and HDPE at 288 h. We used Nylon coupons to determine the kinetic parameters of the planktonic and biofilm phases of P. salmonis. The two strains had similar latencies in the planktonic phase; however, LF-89 maximum growth was 2.5 orders of magnitude higher (Log cell ml-1 ). Additionally, LF-89 had a specified growth rate (µmax) of 0.0177 ± 0.006 h-1 and a generation time of 39.2 h. This study contributes to a deeper understanding of the biofilm formation by P. salmonis and elucidates the impact of the biofilm on aquaculture systems.

Piscirickettsia salmonis Produces a N-Acetyl-L-Homoserine Lactone as a Bacterial Quorum Sensing System-Related Molecule.

Piscirickettsia salmonis is the etiological agent of piscirickettsiosis, the most prevalent disease in salmonid species in Chilean salmonids farms. Many bacteria produce N-acyl-homoserine lactones (AHLs) as a quorum-sensing signal molecule to regulate gene expression in a cell density-dependent manner, and thus modulate physiological characteristics and several bacterial mechanisms. In this study, a fluorescent biosensor system method and gas chromatography-tandem mass spectrometry (GC/MS) were combined to detect AHLs produced by P. salmonis. These analyses revealed an emitted fluorescence signal when the biosensor P. putida EL106 (RPL4cep) was co-cultured with both, P. salmonis LF-89 type strain and an EM-90-like strain Ps007, respectively. Furthermore, the production of an AHL-type molecule was confirmed by GC/MS by both P. salmonis strains, which identified the presence of a N-acetyl-L-homoserine Lactone in the supernatant extract. However, It is suggested that an alternate pathway could synthesizes AHLs, which should be address in future experiments in order to elucidate this important bacterial process. To the best of our knowledge, the present report is the first to describe the type of AHLs produced by P. salmonis.

Molecular weight distribution of the recalcitrant organic matter contained in kraft mill effluents and the identification of microbial consortia responsible for an anaerobic biodegradable fraction.

The objective of this research was to evaluate the distribution of the molecular weights of the recalcitrant organic matter contained in kraft mill effluents and identify microbial consortia responsible for an anaerobic biodegradable fraction. As a result, the average removal efficiencies of chemical organic demand (COD) and biological oxygen demand (BOD5) during the entire period of operation were 28% and 53%, respectively. The non-biodegradable organic matter was detected at molecular weights less than 1000 Da. However, most of the organic matter was in the molecular weight fraction higher than 10000 Da with 32 ± 11.6% COD as well as color (42.3 ± 8.7%), total phenolic compounds (35.9 ± 7.9%) and adsorbable organic compounds (AOX) (13.0 ± 2.7%). Methanogenic acetoclastic archaea of the genera Methanomethylovorans and Methanosarcina were found in the surface and middle zones of the reactor. Moreover, Methanosaeta and Methanolinea were identified in the low zone of the reactor. In all zones of the reactor, Desulfomicrobium and Desulfovibrio were found to be the most dominant genera of sulfate-reducing bacteria (SRB).

Trickling filter technology for biotreatment of nitrogenous compounds emitted in exhaust gases from fishmeal plants.

Odour emissions are a major environmental issue associated with fishmeal production. Laboratory-scale biotrickling filters (BTFs) were inoculated with microbial consortia derived from sewage sludge, with the goal to study the biotreatment of low-loads of methylamines and ammonia that are main components of odorous exhaust gases produced by fishmeal processing plants. A BTF packed with ceramic rings was subjected to a real fishmeal plant emission containing trimethylamine (TMA), dimethylamine (DMA) and monomethylamine (MMA). The highest elimination capacities (ECs) obtained were 372 mg TMA m-3 h-1, 5.518 mg DMA m-3 h-1 and 1.038 mg MMA m-3 h-1, with maximal removal efficiencies of 92% (TMA), 83% (DMA) and 95% (MMA) after 30 days operation. In a different experiment, a polyurethane foam packing was employed to treat ammonia (NH3) at low inlet loads, reaching an EC of 47.19 mg N m-3 h-1 with 99.8% efficiency (inlet load of 47.27 mg N m-3 h-1). Likewise, the microbial community of the polyurethane-associated biofilm was diverse and stable during operation. These results suggested that elimination of volatile amino-compounds using BTFs inoculated with a methylotrophic microbial consortium holds potential for odour removal. In addition, sequencing analysis of 16S rDNA gene fragments allowed the identification of heterotrophic ammonia-oxidizing bacteria that are promising candidates to effectively maintain ammonia elimination in a biotreatment operation of nitrogenous compounds present in exhaust gases from fishmeal facilities.

Potential methane production and molecular characterization of bacterial and archaeal communities in a horizontal subsurface flow constructed wetland under cold and warm seasons.

Organic matter removal in a horizontal subsurface flow constructed wetland (HSSF) treating wastewater is associated with the presence of bacteria and archaea. These organisms perform anaerobic microbial processes such as methanogenesis, which can lead to methane emissions. The aim of this study was to evaluate methane production and characterize the bacterial and archaeal communities found in HSSFs treating secondary urban wastewater during cold and warm seasons. The pilot system used in this study corresponds to four HSSFs, two planted with Phragmites australis (HSSF-Phr) and two planted with Schoenoplectus californicus (HSSF-Sch), the monitoring was carried out for 1335 days. Removal efficiencies for organic matter (biological and chemical oxygen demand) and total and volatile suspended solids were evaluated in each HSSF. Moreover, biomass from each HSSF was sampled during warm and cold season, and methane productions determined by Specific Methanogenic Activity assays(maximum) (SMAm). In the same samples, the quantification and identification of bacteria and archaea were performed. The results showed that the degradation of organic matter (53-67% BOD5 and 51-62% COD) and suspended solids (85-93%) was not influenced by seasonal conditions or plant species. Potential methane production from HSSF-Sch was between 20 and 51% higher than from HSSF-Phr. Moreover, potential methane production during warm season was 3.4-42% higher than during cold season. The quantification of microorganisms in HSSFs, determined greater development of bacteria (38%) and archaea (50-57%) during the warm season. In addition, the species Schoenoplectus californicus has a larger number of bacteria (4-48%) and archaea (34-43%) than Phragmites australis. The identification of microorganisms evidenced the sequences associated with bacteria belong mainly to Firmicutes (42%), Proteobacteria (33%) and Bacteroidetes (25%). The archaea were represented primarily by Methanosarcinales, specifically Methanosaeta (75%) and Methanosarcina (16%). The community structure of the methanogenic archaea in HSSFs did not change throughout the seasons or plant species.

Characterization of a hyperthermophilic sulphur-oxidizing biofilm produced by archaea isolated from a hot spring

Background: Sulphur-oxidizing microorganisms are widely used in the biofiltration of total reduced sulphur compounds (odorous and neurotoxic) produced by industries such as the cellulose and petrochemical industries, which include high-temperature process steps. Some hyperthermophilic microorganisms have the capability to oxidize these compounds at high temperatures (N60°C), and archaea of this group, for example, Sulfolobus metallicus, are commonly used in biofiltration technology. Results: In this study, a hyperthermophilic sulphur-oxidizing strain of archaea was isolated from a hot spring (Chillán, Chile) and designated as M1. It was identified as archaea of the genus Sulfolobus (99% homology with S. solfataricus 16S rDNA). Biofilms of this culture grown on polyethylene rings showed an elemental sulphur oxidation rate of 95.15 ± 15.39 mg S l-1 d-1, higher than the rate exhibited by the biofilm of the sulphur-oxidizing archaea S. metallicus (56.8 ± 10.91 mg l-1 d-1). Conclusions: The results suggest that the culture M1 is useful for the biofiltration of total reduced sulphur gases at high temperatures and for other biotechnological applications.

Microbial life in a fjord: metagenomic analysis of a microbial mat in Chilean patagonia.

The current study describes the taxonomic and functional composition of metagenomic sequences obtained from a filamentous microbial mat isolated from the Comau fjord, located in the northernmost part of the Chilean Patagonia. The taxonomic composition of the microbial community showed a high proportion of members of the Gammaproteobacteria, including a high number of sequences that were recruited to the genomes of Moritella marina MP-1 and Colwelliapsycherythraea 34H, suggesting the presence of populations related to these two psychrophilic bacterial species. Functional analysis of the community indicated a high proportion of genes coding for the transport and metabolism of amino acids, as well as in energy production. Among the energy production functions, we found protein-coding genes for sulfate and nitrate reduction, both processes associated with Gammaproteobacteria-related sequences. This report provides the first examination of the taxonomic composition and genetic diversity associated with these conspicuous microbial mat communities and provides a framework for future microbial studies in the Comau fjord.

Thiosulphate oxidation by Thiobacillus thioparus and Halothiobacillus neapolitanus strains isolated from the petrochemical industry

Sulphur Oxidizing Bacteria (SOB) is a group of microorganisms widely used for the biofiltration of Total Reduced Sulphur compounds (TRS). TRS are bad smelling compounds with neurotoxic activity which are produced by different industries (cellulose, petrochemical). Thiobacillus thioparus has the capability to oxidize organic TRS, and strains of this bacterium are commonly used for TRS biofiltration technology. In this study, two thiosulphate oxidizing strains were isolated from a petrochemical plant (ENAP BioBio, Chile). They were subjected to molecular analysis by real time PCR using specific primers for T. thioparus. rDNA16S were sequenced using universal primers and their corresponding thiosulphate activities were compared with the reference strain T. thioparus ATCC 10801 in batch standard conditions. Real time PCR and 16S rDNA sequencing showed that one of the isolated strains belonged to the Thiobacillus branch. This strain degrades thiosulphate with a similar activity profile to that shown by the ATCC 10801 strain, but with less growth, making it useful in biofiltration.

Biofilm growth kinetics of a monomethylamine producing Alphaproteobacteria strain isolated from an anaerobic reactor.

Industrial fishing effluents are characterized by high loads of protein and sulfate that stimulate the activity of proteolytic and sulfate reducing bacteria during anaerobic digestion. Their metabolic products (NH3 and H2S respectively) have a well-known detrimental effect on the activity of methanogens. Since methylamine is a carbon source used by methylaminotrophic methane producing archaea (mMPA) but not by sulfate reducing bacteria (SRB), enriched mMPA anaerobic biofilms have been developed on ceramics. We propose that methylated amines could be produced in the biofilm by using betaine, a known precursor of methylamine, as a carbon and energy source. We isolated an anaerobic betainotrophic methylaminogenic bacterial strain (bMB) from an anaerobic bioreactor, using betaine as the only carbon and energy source. This strain was identified by a standard biochemical test (API 20NE), cloning, and 16S rDNA sequencing. bMB biofilm structure and biofilm growth kinetic parameters were determined by means of scanning electron microscopy (SEM), and the Gompertz growth model, respectively. Monomethylamine production was determined by infrared spectroscopy and by high pressure liquid chromatography. The isolated bMB strain was determined as Stappia stellulata (Proteobacteria phylum). It was able to form biofilm on ceramics and its kinetic growth parameters resulted in: maximum biofilm bacterial count (A) of 6.25 x 10(8) UFC/cm(2) and maximum specific growth rate (mu(m)) of 0.0221/h. Production of monomethylamine was about 4.027 atogram/cell/day (at/cell/day) after 15 days of incubation in biofilms. This study confirms the adhesion capacity of this bMB strain on ceramic supports, assuring that monomethylamine production in biofilms could be enriched with mMPA that use monomethylamine.