Nagappa Ramaiah

Chromium (Cr) pollution is an emerging environmental problem. The present study was carried out to isolate Cr-resistant bacteria and characterize their Cr detoxification and resistance ability. Bacteria screened by exposure to chromate (Cr⁶⁺) were isolated from Mandovi estuary Goa, India. Two isolates expressed high resistance to Cr⁶⁺ (MIC ≥ 300 µg mL⁻¹), Cr³⁺ (MIC ≥ 900 µg mL⁻¹), other toxic heavy metals and displayed a pattern of resistance to cephalosporins and ß-lactams. Biochemical and 16 S rRNA gene sequence analysis indicated that both isolates tested belonged to the Staphylococcus genus and were closely related to S. saprophyticus and S. arlettae. Designated as strains NIOER176 and NIOER324, batch experiments demonstrated that both removed 100% of 20 and 50 µg mL⁻¹ Cr⁶⁺ within 4 and 10 days, respectively. The rate of reduction in both peaked at 0.260 µg mL⁻¹ h⁻¹. ATP-binding cassette (ABC) transporter gene involved in transport of a variety of substrates including efflux of toxicants was present in strain NIOER176. Through SDS-PAGE analysis, whole-cell proteins extracted from both strains indicated chromium-induced specific induction and up-regulation of 24 and 40 kDa proteins. Since bacterial ability to ameliorate Cr⁶⁺ is of practical significance, these findings demonstrate strong potential of some estuarine bacteria to detoxify Cr⁶⁺ even when its concentrations far exceed the concentrations reported from many hazardous effluents and chromium contaminated natural habitats. Such potential of salt tolerant bacteria would help in Cr⁶⁺ bioremediation efforts.

The potential of the marine microalgal isolate Phormidium sp. to detoxify/reduce various pollutants from a highly hazardous mixed industrial effluent (MIE) is evaluated. The microalgae are grown in the MIE at 30% (v/v%) strength for 21 days under controlled environmental conditions. The reductions/changes in concentrations of biochemical oxygen demand (BOD), chemical oxygen demand (COD), nitrate, nitrite, total phosphate, and toxic metals (chromium, nickel, manganese, copper) are monitored. Notably, the isolate grew quite well, reducing the levels of BOD by 93% and those of COD by 71% in 6 to 9 days. Concentrations of other measured contaminants are also reduced to within the permissible limits of discharge. Higher rates of hatchability in Artemia salina cysts and survivability of its hatchlings are observed in the algae‐treated wastewater than in the untreated wastewater, indicating safe discharge of the wastewater. These observations are valuable, as they indicate that many environmental strains of microalgae possess the potential to detoxify certain hazardous MIEs and can adequately improve the quality of water for reuse/discharge, thereby safeguarding our natural environments from the detrimental effects of direct disposal.

Monsoonal upwelling along south and central west coast of India leads to intense biological productivity. As a consequence of excess organic matter production following upwelling during June–October and low dissolved oxygen in the upwelled waters, denitrification occurs in the near shore waters. Implicitly, these processes ought to bring alterations in microbial communities. Therefore, diversity and community structure of bacteria from subsurface layers of a tropical region experiencing intense seasonal lows of oxygen were analyzed through sequencing of 16S rRNA gene clones. The overall diversity was more during hypoxic period of Fall intermonsoon (FIM, October) compared either to Spring intermonsoon (SIM, March) or summer monsoon (SuM, June) months. As many as 14 lineages of bacterial domains: Gammaproteobacteria (37%), Alphaproteobacteria (21%), Cyanobacteria (20%), Deltaproteobacteria (3%), Firmicutes (3%), Betaproteobacteria (2%), Acidobacteria (2%), Actinobacteria (7%), Marinimicrobia (2%), Bacteroidetes (1%), Verrucomicrobia (1%), Planctomycetes (0.4%), Chloroflexi (0.2%) and Omnitrophica bacterium (0.2%), were recognized from our coastal location. Notably, sequences of Planctomycetes, Chloroflexi and Omnitrophica bacterium were found exclusively during FIM. A generally higher representation of sequences of Betaproteobacteria during SuM and of Actinobacteria and Firmicutes during SIM was discernible. This study is thus useful to recognize that microbial community might undergo strong temporal shifts in the monsoon affected tropical coastal ecosystems.

Liquid Chromatography–Mass Spectrometry-Quadrupole Time of Flight (LC/MS QToF) protein profiling of marine-derived Staphyloccous gallinarum NIOMR8 was carried out to evaluate proteins conferring chromate (Cr⁶⁺) resistance and possible metabolic pathways that were altered as a result. Expressional (up or down-regulation) responses to varying Cr⁶⁺ (0, 50, 100, 150, and 200 µg mL^− 1) concentrations varied, with as many as 346 proteins identified. Most number of proteins—their numbers in parentheses—were up-regulated when grown in medium with 50 µg mL^− 1 (162) and, down-regulated in medium with 100 (281) or 200 µg mL^− 1 Cr⁶⁺ (280). Among these, eight proteins were commonly up-regulated, while 58 were commonly down-regulated across all conditions of Cr⁶⁺. Expression of protein moieties in metabolic pathways like translation (38), transcription (14), replication (18) and repair (4), metabolism of carbohydrates (26), amino acids (27), nucleotides (17), and membrane transport (21) was evidenced. Up-regulation patterns suggest that reduction of molecular oxygen (5), DNA repair (4) and peptide misfolding (7) were the potential protective mechanisms employed to counter Cr⁶⁺ stress. Additionally, proteins associated with biofilm and cell wall biogenesis highlight their hypothetical involvement in toxicity tolerance. Results also indicate that at higher concentrations of Cr⁶⁺, down-regulation of functional proteins impedes normal cellular functions.

This study evaluated the bioremediation potential of two marine microalgae Chlorella sp. and Phormidium sp., both individually and in consortium, to reduce various pollutants in tannery wastewater (TW). The microalgae were grown in hazardous 100% TW for 20 days, and the reductions in biochemical oxygen demand (BOD), chemical oxygen demand (COD), total nitrogen (TN), total phosphorous (TP), chromium (Cr) and total dissolved solids (TDS) of the wastewater monitored periodically. Both marine isolates reduced the BOD and COD by ≥90% in the consortium and by over 80% individually. Concentrations of TN and TP were reduced by 91.16% and 88%, respectively, by the consortium. Removal/biosorption efficiencies for chromium ranged from 90.17–94.45%. Notably, the TDS, the most difficult to deal with, were reduced by >50% within 20 days by the consortium. The novel consortium developed in this study reduced most of the ecologically harmful components in the TW to within the permissible limits of discharge in about 5 to 15 days of treatment. Thus, both the tested marine strains of Chlorella and Phormidium sp. are promising for bioremediating/detoxifying TW and adequately improve the water quality for safe discharge into open water bodies, in particular when used as a consortium.

Marine bacteria possess a wide variety of bioremediation potential which is beneficial environmentally and economically. In this study, bacterial isolates from marine waters were screened for tolerance and growth in high concentrations of chromate (Cr⁶⁺). Two isolates, capable of tolerating Cr⁶⁺ concentrations 300 µg mL⁻¹ or higher, and found to completely reduce 20 µg mL⁻¹ Cr⁶⁺ were grown in Cr⁶⁺ (50 and 100 mg kg⁻¹) spiked garden soil. Notably, both facilitated normal germination and growth of mung (Vigna radiata) seeds, which could hardly germinate in Cr⁶⁺ spiked garden soil without either of these bacteria. In fact, large percent of mung seeds failed to sprout in the Cr⁶⁺ spiked garden soil and could not grow any further. Apparently, chromate detoxification by marine bacterial isolates and the ability of mung plants to deal with the reduced form appear to work complementarily. This study provides an insight into marine bacterial abilities with respect to chromium and potential applications in promoting growth of leguminous plants-similar to mung in particular-in Cr⁶⁺ contaminated soil.

Ocean iron fertilization is an approach to increase CO2 sequestration. The Indo-German iron fertilization experiment “LOHAFEX” was carried out in the Southern Ocean surrounding Antarctica in 2009 to monitor changes in bacterial community structure following iron fertilization-induced phytoplankton bloom of the seawater from different depths. 16S rRNA gene libraries were constructed using metagenomic DNA from seawater prior to and after iron fertilization and the clones were sequenced for identification of the major bacterial groups present and for phylogenetic analyses. A total of 4439 clones of 16S rRNA genes from ten 16S rRNA gene libraries were sequenced. More than 97.35% of the sequences represented four bacterial lineages i.e. Alphaproteobacteria, Gammaproteobacteria, Bacteroidetes and Firmicutes and confirmed their role in scavenging of phytoplankton blooms induced following iron fertilization. The present study demonstrates the response of Firmicutes due to Iron fertilization which was not observed in previous southern ocean Iron fertilization studies. In addition, this study identifies three unique phylogenetic clusters LOHAFEX Cluster 1 (affiliated to Bacteroidetes), 2 and 3 (affiliated to Firmicutes) which were not detected in any of the earlier studies on iron fertilization. The relative abundance of these clusters in response to iron fertilization was different. The increase in abundance of LOHAFEX Cluster 2 and Papillibacter sp. another dominant Firmicutes may imply a role in phytoplankton degradation. Disappearance of LOHAFEX Cluster 3 and other bacterial genera after iron fertilization may imply conditions not conducive for their survival. It is hypothesised that heterotrophic bacterial abundance in the Southern Ocean would depend on their ability to utilise algal exudates, decaying algal biomass and other nutrients thus resulting in a dynamic bacterial succession of distinct genera.

Elucidation, through molecular analyses, of bacterial afflictions in commercially important aquaculture‐reared shrimps is pivotal for the prevention and/or control of disease outbreaks. In this study, we examined the phylogenetic relatedness and compared the possible immune‐related functional roles of both translationally controlled tumour protein (TCTP) and ferritin genes with previous studies. Both TCTP and ferritin genes were substantially upregulated in the Indian white prawn, Fenneropenaeus indicus (H. Milne Edwards), post‐larvae following bath challenge with the virulent strain of bacteria, Vibrio harveyi D3. Full‐length cloning of these genes by rapid amplification of complementary DNA ends ‐polymerase chain reaction (RACE‐PCR) yielded 727‐base pair (bp)‐long TCTP and 1212‐bp‐long ferritin gene sequences. Their open reading frames (ORFs) were 507 and 510 bp, respectively. The TCTP‐ORF coded for 168 amino acids with three substitutions at positions 37, 141, 155, and the ferritin ORF coded for 170 amino acids with no species‐specific substitutions. Phylogenetic analysis suggested the closest relatedness of both TCTP and ferritin from F. indicus to Chinese white prawn, Fenneropenaeus chinensis (Osbeck). In addition to reporting the full‐length sequences of these immune‐relevant genes, this study highlighted their conserved natures, which perhaps make them important defence‐related proteins in the innate immune system of F. indicus.