Isolation of a Dissimilatory Iodate-Reducing Aromatoleum sp. From a Freshwater Creek in the San Francisco Bay Area

Recent reports of dissimilatory iodate-reducing microorganisms (DIRM) have arisen from studies of bacteria in marine environments. These studies described the physiology and distribution of DIRM while also demonstrating their presence in iodine-rich marine environments. We posited that despite lower iodine concentrations, terrestrial and freshwater ecosystems should also harbor DIRM. We established numerous enrichments from coastal and freshwater environments that actively remove amended iodate. We describe the physiology and genome of a new DIRM isolate, Aromatoleum toluclasticum sp. TC-10, emerging from a freshwater creek microcosm. Like other DIRM, A. toluclasticum sp. TC-10 couples acetate oxidation to iodate reduction with a concomitant increase in the OD600. Our results indicate that A. toluclasticum sp. TC-10 performs dissimilatory iodate reduction (DIR) using the recently described iodate reductase (Idr). We provide further evidence of horizontal gene transfer of the idr genes by demonstrating the lack of Idr in the closely related (99.93% 16S rDNA sequence identity) A. toluclasticum sp. MF63 and describe the heterogeneity of the accessory proteins associated with the iodate reduction island (IRI). These observations provide additional evidence that DIR is a horizontally acquired metabolism with broad environmental distribution beyond exclusively marine environments.

Isolation and Characterization of Bacillus Sp. Capable of Degradating Alkali Lignin

Alkali lignin-degrading Bacillus were isolated from forest soils in China and were identified as Bacillus subtilis TR-03 and Bacillus cereus TR-25 by 16S rDNA sequence analysis. Wherein TR-03 displayed optimal 26.72% alkali lignin (2 g/L) degradation at 7 days and 71.23% of Azure-B (0.01%) decolorization at 36 h of cultivation at 37°C. Ligninolytic enzyme analysis revealed that TR-03 was capable of depolymerizing alkali lignin effectively by the producing of lignin peroxidase and laccase, wherein higher laccase activity was cell-associated. At last, the physical and chemical changes of lignin via SEM and FTIR analysis was further observed to prove the lignin degradation by Bacillus subtilis TR-03.

Characterization of the Bacterial Community of Rumen in Dairy Cows with Laminitis

Laminitis is the inflammation of the lamella, and it has caused great economic loss to the dairy industry and attracted wide attention around the world. In recent years, microbiota are considered to play a significant role in various diseases processes. Therefore, our study aimed to explore the characteristics of ruminal microbiota in laminitis cows. The serum of bovines with or without laminitis was collected to detect concentrations of lipopolysaccharide (LPS), lactic acid, and histamine, and ruminal fluid was collected for 16S rDNA sequence analysis. The results showed a significant increase in LPS and lactic acid levels in the laminitis group compared to the control group cows. In addition, a higher abundance of Candidatus Saccharimonas, Saccharofermentans, Erysipelotrichaceae UCG-009 genus, Acetobacter pasteurianus, Clostridium papyrosolvens, Ruminococcaceae bacterium AE2021, Porphyromonas crevioricanis, Pseudomonas boreopolis, Pseudomonas psychrotolerans, Rothia nasimurium, and Rothia pickettii was detected in the rumen fluid of laminitis bovines. In conclusion, this article confirms that there are differences in rumen microbiota between healthy and laminitis bovines. The elevated abundance of bacteria that enrich acid-enhancing metabolites, as well as increase the concentration of lactic acid and LPS, could be harmful factors to bovines and increase the risk of laminitis.

Isolation, identification, and biological characteristics of Clostridium sartagoforme from rabbit

In order to develop microbial additives for rabbit feed, a spore-forming bacteria was isolated from the feces of Hyla rabbit using reinforced clostridium medium (RCM). The 16S rDNA sequence of the bacterium was subjected to pairwise sequence alignment using BLAST; the colony morphology, and physiological, biochemical, and stress resistance were studied. The results showed that the bacterium was Clostridium sartagoforme, a gram positive anaerobe, which can produce spores. The colony diameter was 0.5 mm—2.5 mm, the diameter of the bacteria was 0.5 μm—1.0 μm × 2.0 μm—6.3 μm, and the spore diameter was 1 μm—1.2 μm × 1 μm—1.2 μm. C. sartagoforme can utilize various sugars and alcohols such as fructose, galactose, sorbitol, and inositol. It secreted cellulase into the extracellular environment to form a transparent hydrolysis circle in Congo red medium, it could not liquify gelatin, and the lysine decarboxylase reaction was positive. In liquid medium it entered the stable growth period after 9 h of inoculation. Additionally, it had good stress resistance with a survival rate that exceeded 53% after gastric juice (pH 2.5) treatment for 3 h, it grew in a medium with a bile salt concentration of 0.3%, and the survival rate exceeded 85% after 10 minutes at 80°C. Moreover, animal testing indicated that this strain has no adverse effects on the morbidity and mortality of rabbits. In summary, C. sartagoforme XN-T4 was isolated from rabbit feces. This bacterium has good resistance to stress, can decompose a variety of monosaccharides and polysaccharides including cellulose, which is relatively harmless for animal health.

Characterization of a New L-Glutaminase Produced by Achromobacter xylosoxidans RSHG1, Isolated from an Expired Hydrolyzed L-Glutamine Sample

As significant biocatalyst, L-glutaminases find potential applications in various fields, from nourishment to the pharmaceutical industry. Anticancer activity and flavor enhancement are the most promising applications of L-glutaminases. In this study, L-glutaminase was isolated and purified from an old glutamine sample. A selected bacterial isolate was characterized taxonomically by morphological characters, biochemical testing and 16S rDNA sequence homology testing. The taxonomical characterization of the isolate identified it as Achromobacter xylosoxidans strain RSHG1. The isolate showed maximum enzyme production at 30 °C, pH 9, with Sorbitol as a carbon source and L-Glutamine as a nitrogen and inducer source. L-Glutaminsae was purified by using column chromatography on a Sephadex G-75. The enzyme has a molecular weight of 40 KDa, pH optimal 7 and is stable in the pH range of 6–8. The optimum temperature for the catalyst was 40 °C and stable at 35–50 °C. The kinetic studies of the purified L-glutaminase exhibited Km and Vmax of 0.236 mM and 443.8 U/mg, respectively. L-Glutaminase activity was increased when incubated with 20 mM CaCl2, BaCl2, ZnSO4, KCl, MgSO4 and NaCl, whereas EDTA, CoCl2, HgCl, ZnSO4 and FeSO4 decreased the activity of the enzyme. The addition of 8% NaCl enhanced the glutaminase activity. L-Glutaminase immobilized on 3.6% agar was stable for up to 3 weeks.

Isolation and identification of fenobucarb degrading bacteria from Pangalengan farm land

The long term excessive use of pesticides can lead to their residues accumulation in the soils. Soil microbes were considered to convert the residues into harmless compounds, however the indigenous soil microbes having those beneficial properties are limited. Therefore this study aimed to isolate, select and identify the fenobucarb insecticide-degrading bacteria from agricultural soils. The soil samples were collected from the vegetable fields in Pangalengan, West Java, Indonesia. Isolation of the bacteria was conducted using Nitrate Mineral Salt Agar suplemented by 100 ppm of a fenobucarb. The bacteria isolates were selected based on its hypersensitive response, haemolytic activity, and its ability to degrade fenobucarb. The selected isolates was identified base on sequences of 16S rRNA gene. Twenty nine bacteria were isolated from four soil samples and 23 of the isolates were not potentially phytopathogenic and non haemolytic. The best three isolates that could degrade 94.2%, 94.5% and 95.47% fenobucarb residue are B41, B54 and B83 isolates, respectively. The 16S rDNA Sequence analysis showed that B41 and B83 isolates have 100% similarity to Bacillus thuringiensis MYBT 18426B54, while B54 isolate has 99% similarity to Bacillus luciferensis LMG 18422. These isolates are potential to be developed as a bioremediation agent.

Decolorization of mixture of UV/ TiO2 pre-treated azo dyes by using developed bacterial consortium – An excellent outcome

The degradation of the mixture of azo dyes by the developed bacterial consortium, photocatalytic process (TiO2/UV) and their combined effects were investigated in this study. The bacteria consortium was developed from waste disposal drains in the local textile dyeing industry. The consortium consists of two different bacteria which were identified as Stenotrophomonas pavanii and Bacillus licheniformis through 16S rDNA sequence alignment. The decolorization efficiency was estimated by spectrophotometry and it was observed that biological and photochemical methods alone could not effectively remove the dyes as the decolorization efficiency was low and the absorption peak in the UV region was not completely removed. After 5 days of incubation at 37°C, pH 7 and a dye concentration of 150 mg/L, the microbial dye degradation reached a decolorization efficiency of more than 55%. Additionally, the UV treatment alone was also able to decolorize the dye less than 20% at 45°C, pH 9 at 150 mg/L of dye. A two-step treatment process, namely, photocatalytic treatment followed by biological degradation, was assessed. Ultraviolet-Visible (UVVis) spectral analysis showed that the combined effects were most efficient in the dye degradation (97-98%) which involved a complex interaction of enzyme activity, biosorption and photocatalytic action. Here we also report the optimization of various operational parameters.

Characterization and Genomic Analysis of an Efficient Dibutyl Phthalate-Degrading Bacterium Microbacterium sp. USTB-Y

A promising bacterial strain for biodegrading dibutyl phthalate (DBP) was successfully isolated from activated sludge and characterized as a potential novel Microbacterium sp. USTB-Y based on 16S rDNA sequence analysis and whole genome average nucleotide identity (ANI). Initial DBP of 50 mg/L could be completely removed by USTB-Y both in mineral salt medium and in DBP artificially contaminated soil within 12 h at the optimal culture conditions of pH 7.5 and 30℃, which indicates that USTB-Y has a strong ability in DBP biodegradation. Phthalic acid (PA) was identified as the end-product of DBP biodegraded by USTB-Y using GC/MS. The draft genome of USTB-Y was sequenced by Illumina NovaSeq, and 29 and 188 genes encoding for putative esterase/carboxylesterase and hydrolase/alpha/beta hydrolase were annotated based on NR (non redundant protein sequence database) analysis, respectively. Gene3781 and gene3780 from strain USTB-Y showed 100% identity with dpeH and mpeH from Microbacterium sp. PAE-1. But no phthalate catabolic gene (pht) cluster was found in the genome of strain USTB-Y. This information will be valuable for obtaining a more holistic understanding on diverse genetic mechanisms of PAEs-biodegrading Microbacterium sp. strains.

The Impacts of Different Biological Treatments on the Transformation of Explosives Waste Contaminated Sludge

The dinitrotoluene isomers 2,4 and 2,6-dinitrotoluene (DNT) represent highly toxic, mutagenic, and carcinogenic compounds used in explosive manufacturing and in commercial production of polyurethane foam. Bioremediation, the use of microbes to degrade residual DNT in industry wastewaters, represents a promising, low cost and environmentally friendly alternative technology to landfilling. In the present study, the effect of different bioremediation strategies on the degradation of DNT in a microcosm-based study was evaluated. Biostimulation of the indigenous microbial community with sulphur phosphate (2.3 g/kg sludge) enhanced DNT transformation (82% transformation, from 300 g/L at Day 0 to 55 g/L in week 6) compared to natural attenuation over the same period at 25 °C. The indigenous microbial activity was found to be capable of transforming the contaminant, with around 70% transformation of DNT occurring over the microcosm study. 16S rDNA sequence analysis revealed that while the original bacterial community was dominated by Gammaproteobacteria (30%), the addition of sulphur phosphate significantly increased the abundance of Betaproteobacteria by the end of the biostimulation treatment, with the bacterial community dominated by Burkholderia (46%) followed by Rhodanobacter, Acidovorax and Pseudomonas. In summary, the results suggest biostimulation as a treatment choice for the remediation of dinitrotoluenes and explosives waste.

Characterisation of multi-metal resistant Serratia sp. GP01 for treatment of effluent from fertilizer industries

The effluent generated from fertilizer plants in Paradeep in the coast of the Bay of Bengal is the major pollutant causing health hazard in the vicinity of the area with respect to plants, animals and microbes. Samples of effluent were found to contain heavy metals (mg L-1): Cr (100), Ni (36.975), Mn (68.673), Pb (20.133), Cu (74.44), Zn (176.716), Hg (5.358) and As (24.287) as analyzed by XRF. Indigenous bacterial strains were screened for chromate and multi-metal resistance to remediate the toxic pollutants. The isolated strain G1 was identified as Serratia sp. through 16S-rDNA sequence homology. Potent strain Serratia sp. GP01 treated with 100 mg L-1 of K2Cr2O7 has shown the efficacy of reducing 69.05 mg L-1 of Cr over 48 h of incubation. Further, presence of chromate reductase gene (ChR) in Serratia sp. confirmed the enzymatic reduction of Cr (VI). SEM-EDX and SEM mapping analysis revealed substantial biosorption of Cr and other heavy metals present in effluent by Serratia sp. GP01. Antioxidant enzymes such as catalase (72.15 U mL-1), SOD (57.14 U mL-1) and peroxidase (62.49 U mL-1) were found to be higher as compared to the control condition. FTIR study also revealed the role of N-H, O-H, C = C, C-H, C-O, C-N, and C = O functional groups of the cell surface of Serratia sp. treated with K2Cr2O7 and effluent from the fertilizer industry. Isolated strain Serratia sp. could be used for the detoxification of Cr (VI) and other heavy metals in fertilizer plant effluent.