24 research outputs found

    Bacterial diversity and community structure of banana rhizosphere in Orang Asli fields and commercial plantations

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    Bacteria play an important roles in the soil ecosystem and in the rhizosphere, they are intricately linked to nutrient content and its accessibility to plants, plant protection and sometimes pathogenicity. Banana grows well in the tropics and it is popularly grown in Orang Asli (OA) (indigenous people) settlements. Banana is also grown in commercial plantations. In traditional planting practices, the OA do not add pesticide nor fertilizer to their crops which are planted for selfsustenance mainly. On the other hand, fertilizer and pesticide are added to commercial banana plantations to maximise yield. Rhizosphere bacteria from the banana plant, Pisang Nipah, grown in OA fields and commercial plantations were identified by clone library construction of the 16S rRNA gene. This was to determine whether farming practices influenced the bacterial community in the banana plant rhizosphere. Acidobacteria, Proteobacteria and Actinobacteria were found in all the soil. Other common phyla found in some soil (but not all) were Nitrospirae, Firmicutes, Bacteroidetes, Chloroflexi, Verrumicrobia, Gemmatimonadetes and Cyanobacteria. The bacterial diversity was a little more diverse in the OA fields than the commercial plantations. The latter had higher contents of nitrogen, phosphorus and potassium. These could have exerted selective pressure to reduce the bacterial diversity in the commercial plantations

    Bioinformatic analysis and purification of glutathione transferase (GST) from Pseudomonas sp. UW4

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    The study aimed at identifying and purifying cytosolic glutathione transferase isoforms expressed in Pseudomonas sp. UW4. Search at UniProt (https://www.uniprot.org/uniprot/), has indicated that there were 20 genes encoding putative glutathione transferases for the microorganism. The molecular weights of the isoforms ranged from 17.6 to 34.06 kDa. SDS-polyacrylamide gel electrophoresis revealed that the GST purified using Sulfobromophthalein-glutathione (BSP) affinity column, resolved into a single band with a low molecular weight (MW) of 16 kDa with the pI value of 6.0. Purified GST was reactive towards ethacrynic acid, 1-chloro-2,4-dinitrobenzene, cumene hydroxide, and hydrogen peroxide, but no detectable activity with Trans-2-octenal, hepta-2,4-dienal and Trans-4-phenyl-3-butene-2-one. This has proven that putative GST possessed peroxidase activity and proposed to be similar to PputUW4_00801 (putative glutathione S-transferase) of Pseudomonas sp. UW4 according to its estimated molecular weight and the pI values obtained experimentally

    Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4

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    The study aimed at identifying and purifying cytosolic glutathione transferase isoforms expressed in Pseudomonas sp. UW4. Search at UniProt (https://www.uniprot.org/uniprot/), has indicated that there were 20 genes encoding putative glutathione transferases for the microorganism. The molecular weights of the isoforms ranged from 17.6 to 34.06 kDa. SDS-polyacrylamide gel electrophoresis revealed that the GST purified using Sulfobromophthalein-glutathione (BSP) affinity column, resolved into a single band with a low molecular weight (MW) of 16 kDa with the pI value of 6.0. Purified GST was reactive towards ethacrynic acid, 1-chloro-2,4-dinitrobenzene, cumene hydroxide, and hydrogen peroxide, but no detectable activity with Trans-2-octenal, hepta-2,4-dienal and Trans-4-phenyl-3-butene-2-one. This has proven that putative GST possessed peroxidase activity and proposed to be similar to PputUW4_00801 (putative glutathione S-transferase) of Pseudomonas sp. UW4 according to its estimated molecular weight and the pI values obtained experimentally

    Robust estimation of bacterial cell count from optical density

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    Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

    Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

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    Funder: European Society of Intensive Care Medicine; doi: http://dx.doi.org/10.13039/501100013347Funder: Flemish Society for Critical Care NursesAbstract: Purpose: Intensive care unit (ICU) patients are particularly susceptible to developing pressure injuries. Epidemiologic data is however unavailable. We aimed to provide an international picture of the extent of pressure injuries and factors associated with ICU-acquired pressure injuries in adult ICU patients. Methods: International 1-day point-prevalence study; follow-up for outcome assessment until hospital discharge (maximum 12 weeks). Factors associated with ICU-acquired pressure injury and hospital mortality were assessed by generalised linear mixed-effects regression analysis. Results: Data from 13,254 patients in 1117 ICUs (90 countries) revealed 6747 pressure injuries; 3997 (59.2%) were ICU-acquired. Overall prevalence was 26.6% (95% confidence interval [CI] 25.9–27.3). ICU-acquired prevalence was 16.2% (95% CI 15.6–16.8). Sacrum (37%) and heels (19.5%) were most affected. Factors independently associated with ICU-acquired pressure injuries were older age, male sex, being underweight, emergency surgery, higher Simplified Acute Physiology Score II, Braden score 3 days, comorbidities (chronic obstructive pulmonary disease, immunodeficiency), organ support (renal replacement, mechanical ventilation on ICU admission), and being in a low or lower-middle income-economy. Gradually increasing associations with mortality were identified for increasing severity of pressure injury: stage I (odds ratio [OR] 1.5; 95% CI 1.2–1.8), stage II (OR 1.6; 95% CI 1.4–1.9), and stage III or worse (OR 2.8; 95% CI 2.3–3.3). Conclusion: Pressure injuries are common in adult ICU patients. ICU-acquired pressure injuries are associated with mainly intrinsic factors and mortality. Optimal care standards, increased awareness, appropriate resource allocation, and further research into optimal prevention are pivotal to tackle this important patient safety threat

    Bacterial diversity and community structure of banana rhizosphere in orang asli fields and commercial plantations

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    Bacteria play an important roles in the soil ecosystem and in the rhizosphere, they are intricately linked to nutrient content and its accessibility to plants, plant protection and sometimes pathogenicity. Banana grows well in the tropics and it is popularly grown in Orang Asli (OA) (indigenous people) settlements. Banana is also grown in commercial plantations. In traditional planting practices, the OA do not add pesticide nor fertilizer to their crops which are planted for selfsustenance mainly. On the other hand, fertilizer and pesticide are added to commercial banana plantations to maximise yield. Rhizosphere bacteria from the banana plant, Pisang Nipah, grown in OA fields and commercial plantations were identified by clone library construction of the 16S rRNA gene. This was to determine whether farming practices influenced the bacterial community in the banana plant rhizosphere. Acidobacteria, Proteobacteria and Actinobacteria were found in all the soil. Other common phyla found in some soil (but not all) were Nitrospirae, Firmicutes, Bacteroidetes, Chloroflexi, Verrumicrobia, Gemmatimonadetes and Cyanobacteria. The bacterial diversity was a little more diverse in the OA fields than the commercial plantations. The latter had higher contents of nitrogen, phosphorus and potassium. These could have exerted selective pressure to reduce the bacterial diversity in the commercial plantations

    Effect of temperature on bacterial community in petroleum hydrocarbon-contaminated and uncontaminated Antarctic soil

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    It is generally accepted that bacterial diversity in a community confers resistance to environmental perturbation. Communities with high bacterial diversity are less likely to be impacted by environmental changes such as warming. As such, hydrocarbon-contaminated Antarctic soil that are typically characterised by low bacterial diversity and highly selective taxonomic composition are expected to be more sensitive to changes in temperature than uncontaminated Antarctic soil. To test this hypothesis, we evaluated the response of bacterial community structure to warming of hydrocarbon-contaminated and uncontaminated soil collected from Casey Station, Windmill Island, East Antarctica by using microcosms incubated at 5, 10 and 15 °C over a period of 12 weeks. Our results showed that shifts occurred in the bacterial community in relation to the incubation temperatures in both the hydrocarbon-contaminated and uncontaminated soil, with a stronger response observed in the contaminated soil. Taxa referred as comprising hydrocarbon-degrading genera such as Rhodococcus, was the most prevalent genus in the contaminated soil after incubation at 15 °C, accounting for approximately 32–50% of the total detected genera. However, there were no significant differences in the selected functional genes, potentially suggesting high levels of metabolic plasticity in the studied soil bacterial communities. Overall, we showed that hydrocarbon contamination in soil might lead to lower bacterial community stability against environmental perturbation such as temperature variation

    Spatial pattern in Antarctica: what can we learn from Antarctic bacterial isolates?

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    A range of small- to moderate-scale studies of patterns in bacterial biodiversity have been conducted in Antarctica over the last two decades, most suggesting strong correlations between the described bacterial communities and elements of local environmental heterogeneity. However, very few of these studies have advanced interpretations in terms of spatially associated patterns, despite increasing evidence of patterns in bacterial biogeography globally. This is likely to be a consequence of restricted sampling coverage, with most studies to date focusing only on a few localities within a specific Antarctic region. Clearly, there is now a need for synthesis over a much larger spatial to consolidate the available data. In this study, we collated Antarctic bacterial culture identities based on the 16S rRNA gene information available in the literature and the GenBank database (n > 2,000 sequences). In contrast to some recent evidence for a distinct Antarctic microbiome, our phylogenetic comparisons show that a majority (~75 %) of Antarctic bacterial isolates were highly similar (≥99 % sequence similarity) to those retrieved from tropical and temperate regions, suggesting widespread distribution of eurythermal mesophiles in Antarctic environments. However, across different Antarctic regions, the dominant bacterial genera exhibit some spatially distinct diversity patterns analogous to those recently proposed for Antarctic terrestrial macroorganisms. Taken together, our results highlight the threat of cross-regional homogenisation in Antarctic biodiversity, and the imperative to include microbiota within the framework of biosecurity measures for Antarctica
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