Multiple controls affect arsenite oxidase gene expression in Herminiimonas arsenicoxydans

<p>Abstract</p> <p>Background</p> <p>Both the speciation and toxicity of arsenic are affected by bacterial transformations, i.e. oxidation, reduction or methylation. These transformations have a major impact on environmental contamination and more particularly on arsenic contamination of drinking water. <it>Herminiimonas arsenicoxydans </it>has been isolated from an arsenic- contaminated environment and has developed various mechanisms for coping with arsenic, including the oxidation of As(III) to As(V) as a detoxification mechanism.</p> <p>Results</p> <p>In the present study, a differential transcriptome analysis was used to identify genes, including arsenite oxidase encoding genes, involved in the response of <it>H. arsenicoxydans </it>to As(III). To get insight into the molecular mechanisms of this enzyme activity, a Tn<it>5 </it>transposon mutagenesis was performed. Transposon insertions resulting in a lack of arsenite oxidase activity disrupted <it>aoxR </it>and <it>aoxS </it>genes, showing that the <it>aox </it>operon transcription is regulated by the AoxRS two-component system. Remarkably, transposon insertions were also identified in <it>rpoN </it>coding for the alternative N sigma factor (σ⁵⁴) of RNA polymerase and in <it>dnaJ </it>coding for the Hsp70 co-chaperone. Western blotting with anti-AoxB antibodies and quantitative RT-PCR experiments allowed us to demonstrate that the <it>rpoN </it>and <it>dnaJ </it>gene products are involved in the control of arsenite oxidase gene expression. Finally, the transcriptional start site of the <it>aoxAB </it>operon was determined using rapid amplification of cDNA ends (RACE) and a putative -12/-24 σ⁵⁴-dependent promoter motif was identified upstream of <it>aoxAB </it>coding sequences.</p> <p>Conclusion</p> <p>These results reveal the existence of novel molecular regulatory processes governing arsenite oxidase expression in <it>H. arsenicoxydans</it>. These data are summarized in a model that functionally integrates arsenite oxidation in the adaptive response to As(III) in this microorganism.</p

Microbial communities and arsenic biogeochemistry at the outflow of an alkaline sulfide-rich hot spring

Alkaline sulfide-rich hot springs provide a unique environment for microbial community and arsenic (As) biogeochemistry. In this study, a representative alkaline sulfide-rich hot spring, Zimeiquan in the Tengchong geothermal area, was chosen to study arsenic geochemistry and microbial community using Illumina MiSeq sequencing. Over 0.26 million 16S rRNA sequence reads were obtained from 5-paired parallel water and sediment samples along the hot spring’s outflow channel. High ratios of As(V)/As(Sum) (total combined arsenate and arsenite concentrations) (0.59–0.78), coupled with high sulfide (up to 5.87 mg/L), were present in the hot spring’s pools, which suggested As(III) oxidation occurred. Along the outflow channel, As(Sum) increased from 5.45 to 13.86 μmol/L, and the combined sulfide and sulfate concentrations increased from 292.02 to 364.28 μmol/L. These increases were primarily attributed to thioarsenic transformation. Temperature, sulfide, As and dissolved oxygen significantly shaped the microbial communities between not only the pools and downstream samples, but also water and sediment samples. Results implied that the upstream Thermocrinis was responsible for the transformation of thioarsenic to As(III) and the downstream Thermus contributed to derived As(III) oxidation. This study improves our understanding of microbially-mediated As transformation in alkaline sulfide-rich hot springs

Development of a home literacy environment questionnaire for Tamil-speaking kindergarten children

Abstract Background The development of emergent literacy skills depends upon the literacy environments and experiences of children at home. Children’s home literacy environment is closely related to the development of various aspects of emergent literacy like letter knowledge, phonological awareness, and vocabulary. Dimensions of home literacy environment, such as physical (literacy) environment, child’s own literacy habits, parental literacy habits, parent-child interaction for language and literacy activities, and parental beliefs about literacy, have been reported in literature. Methods The present study describes the construction of a home literacy environment questionnaire for Tamil-speaking kindergarten children in the Indian context. The various dimensions of home literacy environment were described, and items related to each dimension were listed. Seventeen judges rated each item on a five-point Likert scale ranging from 0 (no fit) to 4 (excellent fit). Their comments/remarks/opinion, specific to an item or dimension, were also taken. The rating responses were analyzed for content validity and internal consistency. Results Content validity index was calculated at item level as well as scale level. The items in each subscale/dimension which had the item level content validity index scores higher than 0.78 were included in the final questionnaire and the other items were rejected. The scale level content validity index scores were higher than 0.90, indicating good content validity. Cronbach’s alpha was calculated as a measure of internal consistency. Cronbach’s alpha values were lower than 0.7 for three domains: physical environment, parent literacy habits, and parental beliefs. Conclusions The questionnaire developed promises to be a useful tool to evaluate the home literacy environment of Tamil children who undergo formal education in English medium schools like in India. The questionnaire developed and presented here can help in collecting reliable data to make informed decisions about children’s (whether typically developing or with developmental disabilities) home literacy environment