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Yesha Shrestha

Publications and source records attributed to Yesha Shrestha.

4 recordsLinked to original sources

Acetylenotrophic and diazotrophic Bradyrhizobium sp. strain I71 from TCE-contaminated soils

Abstract Acetylene (C 2 H 2 ) is a molecule rarely found in nature, with very few known natural sources, but acetylenotrophic microorganisms can use acetylene as their primary carbon and energy source. As of 2018 there were 15 known strains of aerobic and anaerobic acetylenotrophs; however, we hypothesize there may yet be unrecognized diversity of acetylenotrophs in nature. This study expands the known diversity of acetylenotrophs by isolating the aerobic acetylenotroph, Bradyrhizobium sp. strain I71, from trichloroethylene (TCE)-contaminated soils. Strain I71 is a member of the class Alphaproteobacteria and exhibits acetylenotrophic and diazotrophic activities, the only two enzymatic reactions known to transform acetylene. This unique capability in the isolated strain may increase the genus’ economic impact beyond agriculture as acetylenotrophy is closely linked to bioremediation of chlorinated contaminants. Computational analyses indicate that the Bradyrhizobium sp. strain I71 genome contains 522 unique genes compared to close relatives. Moreover, applying a novel hidden Markov model of known acetylene hydratase (AH) enzymes identified a putative AH enzyme. Protein annotation with I-TASSER software predicted the AH from the microbe Syntrophotalea acetylenica as the closest structural and functional analog. Furthermore, the putative AH was flanked by horizontal gene transfer (HGT) elements, like that of AH in anaerobic acetylenotrophs, suggesting an unknown source of acetylene or acetylenic substrate in the environment that is selecting for the presence of AH. Importance The isolation of Bradyrhizobium strain I71 expands the distribution of acetylene-consuming microbes to include a group of economically important microorganisms. Members of Bradyrhizobium are well studied for their abilities to improve plant health and increase crop yields by providing bioavailable nitrogen. Additionally, acetylene-consuming microbes have been shown to work in tandem with other microbes to degrade soil contaminants. Based on genome, cultivation, and protein prediction analysis, the ability to consume acetylene is likely not widespread within the genus Bradyrhizobium . These findings suggest that the suite of phenotypic capabilities of strain I71 may be unique and make it a good candidate for further study in several research avenues.

California

An initial comparison of pesticides and amphibian pathogens between natural and created wetlands in the New Jersey Pinelands, 2014–16

A study conducted by the U.S. Geological Survey, in cooperation with the New Jersey Pinelands Commission and Montclair State University, was designed to compare pesticide concentrations and the presence and prevalence of amphibian pathogens between natural ponds and two types of created wetlands, excavated ponds and stormwater basins, throughout the New Jersey Pinelands. The study described herein is part of a larger study by the New Jersey Pinelands Commission designed to compare the functional equivalency of natural and created wetlands throughout the New Jersey Pinelands. Sites were selected on the basis of land-use classifications within a 500-meter radius around each wetland from a pool of natural ponds, excavated ponds, and stormwater basins determined by the New Jersey Pinelands Commission. Water, bed-sediment, anuran-food, and composite larval-anuran-tissue samples were collected from four reference (minimum land-use effects) and four degraded (maximum land-use effects) sites from each wetland type for a total of 24 ponds or basins throughout the New Jersey Pinelands during 2014–16. Prevalence of Ranavirus was determined on the basis of tail clips collected from 60 individual larval anurans in each wetland, and 10 animals from each wetland also were swabbed for the presence of Batrachochytrium dendrobatidis (Bd). Other constituents measured included turbidity, pH, specific conductance, dissolved oxygen, dissolved organic carbon, percent organic carbon in sediment, and composite larval-anuran lipid content. The amount of altered land (percent agricultural plus percent developed) ranged from 0 to 62.4 percent for the natural ponds, 0 to 63.6 percent for the excavated ponds, and 23.3 to 80.2 percent for the stormwater basins. The herbicides atrazine and metolachlor were observed in 60 and 89 percent of the water samples, respectively. The insecticide bifenthrin was the most frequently detected current-use pesticide (greater than 25 percent of the samples) in bed-sediment, anuran-food, and composite larval-anuran-tissue samples. The legacy insecticide p,p' -DDT and its primary degradates p,p' -DDD and p,p' -DDE were the most frequently detected compounds in bed-sediment and anuran-food samples (32–76 percent in sediment samples and 24–72 percent in anuran-food samples). Significantly, greater numbers of pesticides and higher total pesticide concentrations were observed in stormwater basins than in natural and excavated ponds. Reference wetlands had fewer pesticides and lower total pesticide concentrations compared to degraded wetlands, indicating a positive relation between percent altered land and pesticides throughout the New Jersey Pinelands. Ranavirus was observed in larvae from 4 wetlands, including 1 reference natural pond, 1 degraded natural pond, and 2 degraded stormwater basins, with prevalence ranging from 3 to 43 percent. Bd was detected in swabs from 18 animals and in 4 natural ponds (1 reference and 3 degraded), 3 excavated ponds (all reference), and 2 stormwater basins (1 reference and 1 degraded); however, detection probability was low. In the wetlands with Bd detections, between 10 and 30 percent (between 1 and 3) of the animal’s swabbed tested positive for Bd. Owing to the limited number of positive detections for both Bd and Ranavirus , no statistical comparisons between wetland types and land-use classifications were possible.

New Jersey

Detection of diazotrophy in the acetylene-fermenting anaerobe Pelobacter sp. strain SFB93

Acetylene (C2H2) is a trace constituent of the present Earth's oxidizing atmosphere, reflecting a mixture of terrestrial and marine emissions from anthropogenic, biomass-burning, and unidentified biogenic sources. Fermentation of acetylene was serendipitously discovered during C2H2 block assays of N2O reductase, and Pelobacter acetylenicus was shown to grow on C2H2 via acetylene hydratase (AH). AH is a W-containing, catabolic, low-redox-potential enzyme that, unlike nitrogenase (N2ase), is specific for acetylene. Acetylene fermentation is a rare metabolic process that is well characterized only in P. acetylenicus DSM3246 and DSM3247 and Pelobacter sp. strain SFB93. To better understand the genetic controls for AH activity, we sequenced the genomes of the three acetylene-fermenting Pelobacter strains. Genome assembly and annotation produced three novel genomes containing gene sequences for AH, with two copies being present in SFB93. In addition, gene sequences for all five compulsory genes for iron-molybdenum N2ase were also present in the three genomes, indicating the cooccurrence of two acetylene transformation pathways. Nitrogen fixation growth assays showed that DSM3426 could ferment acetylene in the absence of ammonium, but no ethylene was produced. However, SFB93 degraded acetylene and, in the absence of ammonium, produced ethylene, indicating an active N2ase. Diazotrophic growth was observed under N2 but not in experimental controls incubated under argon. SFB93 exhibits acetylene fermentation and nitrogen fixation, the only known biochemical mechanisms for acetylene transformation. Our results indicate complex interactions between N2ase and AH and suggest novel evolutionary pathways for these relic enzymes from early Earth to modern days.

Applied and Environmental Microbiology