A halophilic Chromohalobacter species from estuarine coastal waters as a detoxifier of manganese, as well as a novel bio-catalyst for synthesis of n-butyl acetate.

Anthropogenic pollution due to ferro-manganese ore transport by barges through the Mandovi estuary in Goa, India is a major environmental concern. In this study a manganese (Mn) tolerant, moderately halophilic Chromohalobacter sp. belonging to the family Halomonadaceae was isolated from the sediments of a solar saltern adjacent to this Mandovi estuary. Using techniques of Atomic absorption spectroscopy, Scanning electron microscopy-Energy dispersive X-ray spectroscopy, Fourier-transform infrared spectroscopy and Atomic Force Microscopy, the Chromohalobacter sp. was explored for its ability to tolerate and immobilize Mn in amended and unamended media with 20% natural salt concentration (w/v). In aqueous media supplemented with 0.1 mM Mn, the Chromohalobacter sp. was capable of sequestering up to 76% Mn with an average immobilization rate of 8 mg Mn /g /day. Growth rate kinetic analysis using Gompertz mathematical functions was found to model the experimental data well. The model inferred that the maximum growth rate of Chromohalobacter sp. was at 10% natural salt concentration (w/v). The Chromohalobacter sp. was further found to be multimetal tolerant showing high tolerance to Iron (Fe), Nickel (Ni) and Cobalt (Co), (each at 4 mM), and tolerated Manganese (Mn) up to 6 mM. Morphologically, the Chromohalobacter sp. was a non-spore forming, Gram negative motile rod (0.726 µ× 1.33 µ). The adaptative mechanism of Chromohalobacter sp. to elevated Mn concentrations (1 mM) resulted in the reduction of its cell size to 0.339 µ× 0.997 µ and the synthesis of an extracellular slime, immobilizing Mn from the liquid phase forming Manganese oxide, as confirmed by Scanning Electron Microscopy. The expression of Mnx genes for manganese oxidation further substantiated the finding. This bacterial synthesized manganese oxide also displayed catalytic activity (∼50% conversion) for the esterification of butan-1-ol with CH3COOH to yield n-butyl acetate. This Chromohalobacter sp. being indigenous to marine salterns, has adapted to high concentrations of heavy metals and high salinities and can withstand this extremely stressed environment, and thus holds a tremendous potential as an environmentally friendly "green bioremediator" of Mn from euryhaline environments. The study also adds to the limited knowledge about metal-microbe interactions in extreme environments. Further, since Chromohalobacter sp. exhibits commendable catalytic activity for the synthesis of n-butyl acetate, it would have several potential industrial applications.

Bacterial response to dynamic metal concentrations in the surface sediments of a solar saltern (Goa, India).

The Ribandar solar saltern, situated adjacent to the Mandovi estuary is influenced by the barge transport of ferromanganese ore to the Mormugao harbour (Arabian Sea). The current study focuses on the distribution of metals and related heterotrophic bacterial populations in the surface sediments (0-10 cm) of the Ribandar salterns (Goa, India) during the salt-making (January to May) and non salt-making seasons (August and November). The concentrations of heavy metals in the sediments ranged from 17.2 ± 2.8 to 26.3 ± 6.7 % Fe; 0.6 ± 0.2 to 0.9 ± 0.2 % Mn; 27.6 ± 7.3 to 51 ± 8.3 ppm Ni; 28.4 ± 8.9 to 35.2 ± 10.6 ppm Co; 44 ± 21.6 to 62.8 ± 23.6 ppm Zn; 0.1 ± 0.01 ppm Cd and 1.7 ± 0.1 to 2.6 ± 0.7 ppm Pb and were much higher than those reported at the same site in a previous study by Kerkar (2004). Hg concentrations were below detection limits. In general, computation of "geoaccumulation index" revealed the sediments as 'uncontaminated to moderately contaminated' with Fe, Mn, Ni, Co, Pb and Zn during the salt-making season. The abundance of metal-tolerant bacteria was comparatively restricted to the salt-making season and was higher than the non salt-making season. Fe-, Mn-, Ni-, Co- and Pb- (200 ppm) tolerant bacteria were retrieved and restricted to the surface sediments (0-5 cm), Cd and Fe being the two most regulatory elements governing bacterial populations in the non salt-making season. However, during the salt-making season, the concentration of Zn was found to be pivotal in regulating the counts of Fe-, Mn- and Ni-tolerant bacteria. In general, the strength of correlation of metals and microbes was higher in the non salt-making season as compared to the salt-making season. This would probably indicate metal-induced limitations in microbial populations in the non salt-making season and the absence of this effect during the salt-making season. In this study, we test the hypothesis that solar salterns behave as ecological sinks with a potential to transform native bacterial populations to metal-resistant strains, in relation to the dynamic changes in the surrounding metal concentrations.