Till Örebro universitet

Arsenic (As) toxicity and diabetes mellitus (DM) are emerging public health concerns worldwide. Although exposure to high levels of As has been associated with DM. whether there is also an association between low and moderate As exposure and DM remains unclear. We explored the dose-dependent association between As exposure levels and hyperglycemia, with special consideration of the impact of demographic variables, in 641 subjects from rural Bangladesh. The total study participants were divided into three groups depending on their levels of exposure to As in drinking water (low, moderate and high exposure groups). Prevalence of hyperglycemia, including impaired glucose tolerance (IGT) and DM was significantly associated with the subjects' drinking water arsenic levels. Almost all exposure metrics (As levels in the subjects' drinking water, hair and nails) showed dose-dependent associations with the risk or hyperglycemia, IGT and DM. Among the variables considered, sex, age, and BMI were found to be associated with higher risk of hyperglycemia. IGT and DM. In sex-stratified analyses, As exposure showed a clearer pattern of dose-dependent risk for hyperglycemia in females than males. Finally, drinking water containing low-to-moderate levels of As (50.01-150 mu g/L) was found to confer a greater risk of hyperglycemia than safe drinking water (As <= 10 mu g/L). Thus the results suggested that As exposure was dose-dependently associated with hyperglycemia, especially in females.

Lactic acid bacteria isolated from unconventional sources are often attractive targets in the quest for obtaining better probiotics. In the present study, 16 members of the genus Lactobacillus, isolated from 3 different sources in western India, viz., plants, fermented foods and beverages, and human feces, were evaluated for their probiotic and bioactive properties. The isolates were closely related to Lactobacillus fermentum, Lactobacillus pentosus, and mainly Lactobacillus plantarum. The isolates were tolerant to bile salt, acidic pH and pancreatin, although pancreatin tolerance was generally low. Cellular extracts of several isolates displayed antioxidant activity, while cell-free supernatants displayed antibacterial activity against human pathogens. Antioxidant activity of Lactobacilli of human origin was higher than those from vegetables or fermented foods and beverages. L. plantarum AG40V prevented spoilage of fresh-cut fruits, vegetables and sprouted mung-beans. Lactobacilli from all sources displayed equal probiotic potential and those of human origin displayed superior antioxidant activity over others.

Food-based exposure to arsenic is a human carcinogen and can severely impact human health resulting in many cancerous diseases and various neurological and vascular disorders. This project is a part of our attempts to develop new varieties of crops for avoiding arsenic contaminated foods. For this purpose, we have previously identified four key genes, and molecular functions of two of these, AtACR2 and AtPCSl, have been studied based on both in silico and in vivo experiments. In the present study, a T-DNA tagged mutant, (SALK-143282C with mutation in AtACR2 gene) of Arabidopsis thaliana was studied for further verification of the function of AtACR2 gene. Semi-quantitative RT-PCR analyses revealed that this mutant exhibits a significantly reduced expression of the AtACR2 gene. When exposed to 100 μM of arsenate (AsV) for three weeks, the mutant plants accumulated arsenic approximately three times higher (778 μg/g d. wt.) than that observed in the control plants (235 μg/g d. wt.). In contrast, when the plants were exposed to 100 μM of arsenite (AsIII), no significant difference in arsenic accumulation was observed between the control and the mutant plants (535 μg/g d. wt. and 498 μg/g d. wt., respectively). Also, when arsenate and arsenite was measured separately either in shoots or roots, significant differences in accumulation of these substances were observed between the mutant and the control plants. These results suggest that AtACR2 gene is involved not only in accumulation of arsenic in plants, but also in conversion of arsenate to arsenite inside the plant cells. © 2017 Institute of Molecular Biology, Slovak Academy of Sciences.

The previously described chromium resistant bacterium, Enterobacter cloacae B2-DHA, was isolated from leather manufacturing tannery landfill in Bangladesh. Here we report the entire genome sequence of this bacterium containing chromium and other heavy metal resistance genes. The genome size and the number of genes, determined by massive parallel sequencing and comparative analysis with other known Enterobacter genomes, are predicted to be 4.22 Mb and 3958, respectively. Nearly 160 of these genes were found to be involved in binding, transport, and catabolism of ions as well as efflux of inorganic and organic compounds. Specifically, the presence of two chromium resistance genes, chrR and chrA was verified by polymerase chain reaction. The outcome of this research highlights the significance of this bacterium in bioremediation of chromium and other toxic metals from the contaminated sources.

In this chapter, arsenic-accumulating and arsenic- transformingbacterial strains that can be employed as a sourcefor cost-effective and eco-friendly bioremediation of arsenicsfrom contaminated environments have been reviewed. Thischapter demonstrates that many naturally occurring bacterialstrains like B1-CDA have the potential for reducing arseniccontent in contaminated sources to safe levels. Therefore,the socioeconomic impact of this kind of microorganisms ishighly significant for those countries, especially in the developingworld, where impoverished families and villages aremost impacted. Therefore, this discovery should be consideredto be the most significant factor in formulating nationalstrategies for effective poverty elimination. Besides humanarsenic contamination, these bacterial strains will also benefitlivestock and native animal species, and the outcome ofthese studies is vital not only for people in arsenic-affectedareas but also for human populations in other countries thathave credible health concerns as a consequence of arseniccontaminatedwater and foods.

We have cloned, characterized and transformed the AtACR2 gene (arsenic reductase 2) of Arabidopsis thaliana into the genome of tobacco (Nicotiana tabacum, var Sumsun). Our results revealed that the transgenic tobacco plants are more tolerant to arsenic than the wild type ones. These plants can grow on culture medium containing 200μM arsenate, whereas the wild type can barely survive under this condition. Furthermore, when exposed to 100μM arsenate for 35days the amount of arsenic accumulated in the shoots of transgenic plants was significantly lower (28μg/g d wt.) than that found in the shoots of non-transgenic controls (40μg/g d wt.). However, the arsenic content in the roots of transgenic plants was significantly higher (2400μg/g d. wt.) than that (2100μg/g d. wt.) observed in roots of wild type plants. We have demonstrated that Arabidopsis thaliana AtACR2 gene is a potential candidate for genetic engineering of plants to develop new crop cultivars that can be grown on arsenic contaminated fields to reduce arsenic content of the soil and can become a source of food containing no arsenic or exhibiting substantially reduced amount of this metalloid.

Pollution control has become a priority task for global regulatory authorities. The framing of regulations, guidelines, and implementation of pollution awareness and control programs has begun at a massive scale. Heavy metals that are one of the most challenging pollutants that affect humans, animals, plants, and the ecosystem health. The sources of different metals and their toxicities are described. Current approaches in bioremediation are addressed along with the challenges posed by them. Furthermore, recent developments in biotechnology that offer novel ways to recover metals from contaminated sites are discussed.

Heavy metal pollutants, discharged into the ecosystem as waste by anthropogenic activities, contaminate drinking water for millions of people and animals in many regions of the world. Long term exposure to these metals, leads to several lethal diseases like cancer, keratosis, gangrene, diabetes, cardio- vascular disorders, etc. Therefore, removal of these pollutants from soil, water and environment is of great importance for human welfare. One of the possible eco-friendly solutions to this problem is the use of microorganisms that can accumulate the heavy metals from the contaminated sources, hence reducing the pollutant contents to a safe level.

In this thesis an arsenic resistant bacterium Lysinibacillus sphaericus B1-CDA, a chromium resistant bacterium Enterobacter cloacae B2-DHA and a nickel resistant bacterium Lysinibacillus sp. BA2 were isolated and studied. The minimum inhibitory concentration values of these isolates are 500 mM sodium arsenate, 5.5 mM potassium chromate and 9 mM nickel chloride, respectively. The time of flight-secondary ion mass spectrometry and inductively coupled plasma-mass spectroscopy analyses revealed that after 120 h of exposure, the intracellular accumulation of arsenic in B1-CDA and chromium in B2-DHA were 5.0 mg/g dwt and 320 μg/g dwt of cell biomass, respectively. However, the arsenic and chromium contents in the liquid medium were reduced to 50% and 81%, respectively. The adsorption values of BA2 when exposed to nickel for 6 h were 238.04 mg of Ni(II) per gram of dead biomass indicating BA2 can reduce nickel content in the solution to 53.89%. Scanning electron micrograph depicted the effect of these metals on cellular morphology of the isolates. The genetic composition of B1-CDA and B2-DHA were studied in detail by sequencing of whole genomes. All genes of B1-CDA and B2-DHA predicted to be associated with resistance to heavy metals were annotated.

The findings in this study accentuate the significance of these bacteria in removing toxic metals from the contaminated sources. The genetic mechanisms of these isolates in absorbing and thus removing toxic metals could be used as vehicles to cope with metal toxicity of the contaminated effluents discharged to the nature by industries and other human activities.

Previously, we reported a chromium-resistant bacterium, Enterobacter cloacae B2-DHA, isolated from the landfills of tannery industries in Bangladesh. Here, we investigated its genetic composition using massively parallel sequencing and comparative analysis with other known Enterobacter genomes. Assembly of the sequencing reads revealed a genome of ~4.21 Mb in size.

Here, we report the genomic sequence and genetic composition of an arsenic resistant bacterium Lysinibacillus sphaericus B1-CDA. Assembly of the sequencing reads revealed that the genome size is ~4.5 Mb encompassing ~80% of the chromosomal DNA.