Richa Shrivastava

Suboptimal chlorine treatment of drinking water leads to selection of multidrug-resistant Pseudomonas aeruginosa

The present study was undertaken to investigate the spectrum of bacteria present in the River Gomti water before and after chlorination for drinking purposes. We observed that the strains of Pseudomonas aeruginosa that survived chlorination on three out of seven occasions were resistant to almost all the antibiotics tested. The chlorine-resistant bacteria had mucoid colonies and grew better at 24 degrees C. All attempts to isolate the plasmid responsible for chlorine resistance were unsuccessful. Laboratory experiments using different strains of the P. aeruginosa in distilled water showed that only the resistant strain survived chlorine treatment at a dose of < or =500 microg/L. Similar results were obtained when water collected from seven different sites on the River Gomti was treated with graded doses of chlorine. At the higher dose of chlorine, all the bacteria died in 30 min, whereas with lower doses all the bacteria survived. The present study underscores the importance of measuring water chlorine concentrations to assure they are sufficiently high to remove pathogenic bacteria from drinking water. To our knowledge, this is the first report in the literature of the selection of multidrug-resistant bacteria by suboptimal chlorine treatment of water.

Various cells of the immune system and intestine differ in their capacity to reduce hexavalent chromium

The cells of the immune system form a strong line of defence against foreign substances. The present study was undertaken to investigate the capacity of different cells of Wistar rats to reduce potentially carcinogenic hexavalent chromium (Cr-VI) into less toxic trivalent chromium in vitro. 5 x 10(6) cells were incubated with 10 or 25 microg ml(-1) of Cr (VI) in the form of K2Cr2O7 at 37 degrees C in the presence of 5% CO2 in air. At various time periods the remaining amount of Cr (VI) was measured and the percentage of Cr (VI) reduced was calculated. Among the single cell suspensions from the splenic cells a peak reduction of 55% was observed with the total spleen cells, 40% with the B-lymphocyte-enriched subpopulation, 10% with T-lymphocytes and 24% with the macrophages. The reduction by splenic and peritoneal macrophages was similar. Total thymocytes reduced 54% of the Cr (VI). Since the most common route of entry of chromium is through drinking water and food, intestinal cells were also investigated. Among the intestinal cells the maximum reduction of 100% (of 10 microg ml(-1)) was observed with the upper villus cells and 72% with the middle villus cells while reduction was the least (4%) with the crypt cells. The reduction in the intestinal loop in situ was 100%. The time taken by each cell type for the peak reduction to Cr (VI) was markedly different. The findings thus show that the capacity of different cells in the body differs vastly in their capacity and time taken to reduce hexavalent chromium. The most efficient handling of Cr (VI) by the intestine, due to the presence of a variety of cells and bacteria, protects the body from its adverse effects.

Interaction of viral proteins with metal ions: role in maintaining the structure and functions of viruses

Abstract Metal ions are integral part of some viral proteins and play an important role in their survival and pathogenesis. Zinc, magnesium and copper are the commonest metal ion that binds with viral proteins. Metal ions participate in maturation of genomic RNA, activation and catalytic mechanisms, reverse transcription, initial integration process and protection of newly synthesized DNA, inhibition of proton translocation (M2 protein), minus- and plus-strand transfer, enhance nucleic acid annealing, activation of transcription, integration of viral DNA into specific sites and act as a chaperone of nucleic acid. Metal ions are also required for nucleocapsid protein-transactivation response (TAR)–RNA interactions. In certain situations more than one metal ion is required e.g. RNA cleavage by RNase H. This review underscores the importance of metal ions in the survival and pathogenesis of a large group of viruses and studies on structural basis for metal binding should prove useful in the early design and development of viral inhibitors.

A Comparative Study on Rat Intestinal Epithelial Cells and Resident Gut Bacteria: (I) Effect of Hexavalent Chromium

Toxicants including heavy metals reaching the intestine following ingestion through food and water primarily interact with an ecosystem of eukaryotic and prokaryotic cells. Gut bacteria having a dynamic interrelationship with intestinal epithelial cells are known to play important and specific metabolic, trophic, and protective functions. The present study was undertaken to compare the effects of hexavalent chromium on rat intestinal epithelial cells and the resident gut bacteria following in vitro and in vivo exposures. The survival rate and viability pattern of two types of cells were comparable. Under in vitro conditions, the gut bacteria were quick to reduce Cr (VI) in early time periods, while, at 30 h time, both types of cells showed similar capacity for the reduction of Cr (VI). Chromium intoxication (10 ppm of Cr (VI) in drinking water for 10 weeks) caused significant decrease in membrane alkaline phosphatase and Ca2 +-Mg2 +-ATPase activities of intestinal epithelial cells as well as of three gut bacteria viz. Escherichia coli, Pseudomonas sp, and Lactobacillus sp. Major structural membrane constituents like carbohydrates and phospholipids also showed significant decline in both types of cells. These findings indicate that 10 ppm and higher Cr concentrations may cause toxic insult, resulting in impaired intestinal functional efficacy. It also implies that the gut bacteria can be used at least for preliminary screening of heavy metals gastrointestinal toxicity.

Effects of chromium on the resident gut bacteria of rat.

The major nonoccupational source of chromium (Cr) for humans is through ingestion with food and water, but its effect on the gut microflora has not been studied. The present study was, therefore, undertaken to investigate the effects of chronic ingestion of potassium dichromate (chromium VI) on the resident gut bacteria of male Wistar rats. A group of rats was kept on drinking water containing 10 ppm chromium VI (Cr [VI]) (called Cr-stressed animals) and the other group was given plain water. After 10 weeks, Lactobacillus, Pseudomonas sp., and Escherichia coli were isolated from the cecum of the rats and various studies were performed. The most significant findings of the present study were the stimulation of growth of facultative gut bacteria from the Cr-stressed rats and the significant increase of growth even in the presence of lower concentrations of Cr. Furthermore, the capacity to reduce Cr (VI) was significantly decreased along with the increased tolerance of the bacteria to Cr (higher minimum inhibitory concentration [MIC] values), which was associated with the development of antibiotic resistance. The effects were most marked with the Pseudomonas sp. and least with the E. coli. The antibiotic resistance developed with the Lactobacillus may be a blessing in disguise, because the bacteria may continue to provide benefits even in patients given antibiotic therapy. The gut bacteria thus provide the first line of defense to the body by converting toxic Cr (VI) to a less toxic Cr (III) and may act as a prebiotic.

Effects of Dengue Virus Infection on the Spleen of Male Mice Given Hexavalent Chromium with Drinking Water

The present study was undertaken to investigate the effects of dengue virus (DV) infection in male mice given drinking water containing 250 ppm Cr (VI) and the normal control male mice given plain water to drink. On the basis of intake of water in 24 h, the average dose of Cr (VI) in each mouse was 14.8 mg/kg. After 3, 6, and 9 weeks of drinking Cr (VI), a set of five mice from each group were inoculated intracerebrally (ic) with a 1000 × LD50 (100 times the lethal dose that kills 50% mice) dose of DV, and the effects on the spleen were studied at the fourth and eightth day postinoculation. It was observed that Cr (VI) drinking and DV infection led to reduction in the weight of the spleen, but the peak reduction was seen in Cr (VI)-fed mice infected with DV, being 30, 34, and 61% at 3, 6, and 9 weeks respectively. A similar response was seen with respect to the cytotoxic activity of spleen homogenates, phagocytic activity of macrophages, and the mitogenic response of spleen cells to concanavalin A from different groups of animals, being most marked (58 to 60%) at the ninth week of Cr (VI) drinking. This shows a summation of adverse effects of DV infection in mice preexposed to Cr (VI).