Draft genome sequence and morphological data of Planifilum fimeticola PLACP1, a thermophilic chloramphenicol-resistant bacterium isolated from thermophilic sludge.

Planifilum is a thermophilic aerobic Actinomyces often found in compost that is suggested to play a primary role in the degradation of organic matter and is a potential antibiotic-resistance gene (ARG)-hosting bacterium during the composting process. Planifilum fimeticola PLACP1 was isolated from thermophilic sludge on a Columbia plate supplemented with chloramphenicol. PLACP1 was Gram-stain-positive with cells longer than 20 µm that branched and intertwined with each other. A draft genome sequence of P. fimeticola PLACP1 was generated using the Illumina NovaSeq system and deposited in the National Center for Biotechnology Information database under the BioProject accession numbers PRJDB17484 and SAMD00736731. The genome sequence comprised 3,395,140 bp, with 57.97 % GC content and 3,368 genes, including 3,267 protein-coding, 6 rRNA, and 56 tRNA genes. Based on the Comprehensive Antibiotic Resistance Database, 237 predicted gene products were related to ARGs, including 44 macrolide antibiotic-related genes (19 %) as the largest group. This dataset will be beneficial for the morphological identification, comparative genomic analyses, and ARG research in the genus Planifilum.

MlrA, an Essential Enzyme for Microcystins and Nodularin on First Step Biodegradation in Microcystin-Degrading Bacteria.

Microcystin-degrading bacteria first degrade microcystins by microcystinase A (MlrA) to cleave the cyclic structure of microcystins at the Adda-Arg site of microcystin-LR, microcystin-RR, and microcystin-YR, but the cleavage of the other microcystins was not clear. In our study, the microcystin-degrading bacterium Sphingopyxis sp. C-1 as wild type and that of mlrA-disrupting mutant, Sphingopyxis sp. CMS01 were used for microcystins biodegradation. The results showed MlrA degraded microcystin-LA, microcystin-LW, microcystin-LY, microcystin-LF, and nodularin. MlrA could cleave the Adda-L-amino acid site.

Biohydrogen production by mixed culture of Megasphaera elsdenii with lactic acid bacteria as Lactate-driven dark fermentation.

Numerous attempts have been made to upscale biohydrogen production via dark fermentation (DF); however, the Achilles' heel of DF, i.e., lactic acid bacteria (LAB) contamination and overgrowth, hinders such upscaling. Key microbes are needed to develop a lactate-driven DF system that can serve as a lactate fermentation platform. In this study, the utility of Megasphaera elsdenii and LAB co-culturing in lactate-driven DF was evaluated. When inoculated simultaneously with LAB or after LAB culture, M. elsdenii achieved a stable hydrogen yield of 0.95-1.49 H2-mol/mol-glucose, approximately half that obtained in pure M. elsdenii cultures. Hydrogen production was maintained even at an initial M. elsdenii-to-LAB cell ratio of one-millionth or less. Moreover, M. elsdenii produced hydrogen via lactate-driven DF from unusable sugars such as xylose or cellobiose. Thus, M. elsdenii could be a Game changer instrumental in unlocking the full potential of DF.

Temperature affects growth, geosmin/2-methylisoborneol production, and gene expression in two cyanobacterial species.

Cyanobacterial blooms accompanied by taste and odor (T&O) compounds affect the recreational function and safe use of drinking water. Geosmin and 2-methylisoborneol (2-MIB) are the most common T&O compounds. In this study, we investigated the effect of temperature on geosmin and 2-MIB production in Dolichospermum smithii and Pseudanabaena foetida var. intermedia. More specifically, transcription of one geosmin synthase gene (geoA) and two 2-MIB synthase genes (mtf and mtc) was explored. Of the three temperatures (15, 25, and 35 °C) tested, the maximum Chl-a content was determined at 25 °C in both D. smithii and P. foetida var. intermedia. The maximum total geosmin concentration (19.82 µg/L) produced by D. smithii was detected at 25 °C. The total 2-MIB concentration (82.5 µg/L) produced by P. foetida var. intermedia was the highest at 35 °C. Besides, the lowest Chl-a content and minimum geosmin/2-MIB concentration were observed at 15 °C. There was a good positive correlation between geosmin/2-MIB concentration and Chl-a content. The expression levels of the geoA, mtf, and mtc genes at 15 °C were significantly higher than those at 25 and 35 °C. The transcription of the mtf and mtc genes in P. foetida var. intermedia was higher at 35 °C than at 25 °C. The results highlight unfavorable temperature can increase the potential of geosmin/2-MIB synthesis from the gene expression level in cyanobacteria. This study could provide basic knowledge of geosmin/2-MIB production by cyanobacteria for better understanding and management of T&O problems in drinking water.

Effects of elevated nitrogen on the growth and geosmin productivity of Dolichospermum smithii.

Geosmin is one of the most common earthy-musty odor compounds, which is mainly produced by cyanobacteria in surface water. Nitrogen (N) is an important factor affecting the growth of cyanobacteria and its secondary metabolites production due to the eutrophication. In this study, we compared the effects of elevated N on the growth and geosmin productivity of Dolichospermum smithii NIES-824 (synonym Anabaena smithii NIES-824), aiming to better understand the mechanisms involved and give an important and fundamental knowledge to solve off-flavor problem. Results show that elevated N concentration promoted more chlorophyll a (Chl-a) production, whereas the geosmin synthesis decreased, revealing a possible competitive correlation between the Chl-a concentration and geosmin production of D. smithii NIES-824. The majority of geosmin (> 90%) was retained intracellularly during the 28 days of cultivation. The qRT-PCR analysis demonstrates that the expression level of the geosmin synthase gene (geoA) was constitutive and decreased at the higher N concentration during the exponential growth phase of cyanobacterial cells. Furthermore, the decrease of geoA expression during the decline phase suggested that geoA transcription was closely related to cell activity and isoprenoid productivity.

Bio-hythane production from cassava residue by two-stage fermentative process with recirculation.

The two-stage hythane fermentation of cassava residue low in protein, rich in iron, and deficient in nickel and cobalt, resulted in failure after long-term operation, showing a radical decrease in methane production along with an increase in volatile fatty acids (VFAs) accumulation in the second stage. Based on the gap between theoretical demand and existing content of nutrients, the effect of their additions on hythane fermentation was validated in the repeated batch experiment and continuous experiment. The proliferation of hydrolysis bacteria, acidogens, and hydrogen producing bacteria and methanogens was guaranteed by sufficient N (0.7g/L), S (30mg/L), Ni (1.0mg/L), and Co (1.0mg/L), and the metabolism of a sustainable hythane fermentation was recovered. In this optimal nutrient combination of above trace elements, the highest hythane yield (426m3 hythane with 27.7% of hydrogen from 1ton of cassava residue) was obtained.

Community analysis of picocyanobacteria in an oligotrophic lake by cloning 16S rRNA gene and 16S rRNA gene amplicon sequencing.

In this study, the picocyanobacterial species composition of Lake Miyagase was examined by analyzing the 16S rRNA gene in a clone library and by amplicon sequencing using a benchtop next-generation sequencer. Five separate samples were analyzed from different days over a ten-month period. In the picocyanobacterial lineage, 9 and 12 OTUs were identified from a clone library and by amplicon sequencing, respectively. Both analyses suggested that a picocyanobacterium related to Synechococcus sp. MW6B4 was dominant in Lake Miyagase. Our findings suggest that 16S rRNA gene amplicon sequencing enables detailed evaluation of picocyanobacteria composition. One OTU identified was found to be a novel cluster that does not group with any of the known freshwater picocyanobacteria.

Development of a 16S rRNA gene primer and PCR-restriction fragment length polymorphism method for rapid detection of members of the genus Megasphaera and species-level identification.

The genus Megasphaera is relevant to the environment, human health and food, and renewable energy for the future. In this study, a primer set was designed for PCR-restriction fragment length polymorphism (RFLP) analyses to detect and identify the members of Megasphaera. Direct detection and identification were achieved for environmental samples and isolates.

Environmental conditions associating microcystins production to Microcystis aeruginosa in a reservoir of Thailand.

Three heptapeptide toxins, microcystin-RR, microcystin-RY and microcystin-LR, which can cause health problems in animals and humans were monitored in Bang Phra Reservoir, Thailand using reversed-phase high performance liquid chromatography. The concentrations of the three toxins in the reservoir varied greatly depending on location and time water samples were collected. Water quality parameters such as light intensity, temperature, pH, dissolved oxygen, suspended solid, chemical oxygen demand, dissolved organic carbon, total nitrogen, total phosphorus, ammonia, nitrate, phosphate, total dissolved nitrogen, total dissolved phosphorus and chlorophyll-a were also measured in parallel with microcystin determinations. Relationships among water quality parameters, toxins and chlorophyll-a were established. Toxin concentration increased in proportion to increases in total phosphorus, fraction of dissolved phosphorus, but was inversely correlated with water pH and total suspended solids. The other measured parameters in the study showed no correlations to toxin level in reservoir water. Significant correlations between chlorophyll-a and suspended solids, phosphate, nitrate and ammonia were observed suggesting that nitrogen and phosphorus are the two major nutrients governing growth of algae in the reservoir. This relationship suggests that algal production as well as toxin concentration are dependant on nutrient levels in the water body, since both measured light intensity and temperature level was favorable for algal growth. A small algal bloom observed in the rainy season of each year (lasting for only a couple of months) paralleled measured increases in toxin concentration, chlorophyll-a, TP and TN in the water column. Toxin level in the water column remain detectable for 3-4 months period following the initiation of algal bloom. Results indicate that major blooms are likely to occur following the raining season which usually occurs near the end of October when runoff would increase nutrient level entering the reservoir. This study also demonstrated that an ongoing field-monitoring program is needed in these lakes and reservoirs for predicting toxic level of microcystin production for use in risk assessment and for alerting the public to potential health hazards. Concentration of toxin in the reservoir can perhaps be controlled by reducing non point source nutrient input within the watershed.