Heavy Metals Induced Modulations in Growth, Physiology, Cellular Viability, and Biofilm Formation of an Identified Bacterial Isolate.

The release of untreated tannery effluents comprising biotoxic heavy metal (HM) compounds into the ecosystem is one of our society's most serious environmental and health issues. After discharge, HM-containing industrial effluents reach agricultural soils and thus negatively affect the soil microbial diversity. Considering these, we assessed the effect of HMs on identified soil beneficial bacteria. Here, the effects of four heavy metals (HMs), viz., chromium (Cr), cadmium (Cd), nickel (Ni), and lead (Pb), on cellular growth, physiology, cell permeability, and biofilm formation of Enterobacter cloacae MC9 (accession no.: MT672587) were evaluated. HMs in a concentration range of 25-200 µg mL-1 were used throughout the study. Among HMs, Cd in general had the maximum detrimental effect on bacterial physiology. With increasing concentrations of HMs, bacterial activities consistently decreased. For instance, 200 µgCr mL-1 concentration greatly and significantly (p ≤ 0.05) reduced the synthesis of indole-3-acetic acid (IAA) by 70% over control. Furthermore, 200 µg mL-1 Cd maximally and significantly (p ≤ 0.05) reduced the synthesis of 2,3-dihydroxybenzoic acid (2,3-DHBA), salicylic acid (SA), 1-aminocyclopropane 1-carboxylate (ACC) deaminase, and extra polymeric substances (EPSs) of E. cloacae MC9 by 80, 81, 77, and 59%, respectively, over control. While assessing the toxic effect of HMs on the P-solubilizing activity of E. cloacae, the toxicity pattern followed the order Cr (mean value = 94.6 µg mL-1) > Cd (mean value = 127.2 µg mL-1) > Pb (mean value = 132.4 µg mL-1) > Ni (mean value = 140.4 µg mL-1). Furthermore, the colony-forming unit (CFU) count (Log10) of strain MC9 was completely inhibited at 150, 175, and 200 µg mL-1 concentrations of Cr and Cd. The confocal laser scanning microscopic (CLSM) analysis of HM-treated bacterial cells showed an increased number of red-colored dead cells as the concentration of HMs increased from 25 to 200 µg mL-1. Likewise, the biofilm formation ability of strain MC9 was maximally (p ≤ 0.05) inhibited at higher concentrations of Cd. In summary, the present investigation undoubtedly suggests that E. cloacae strain MC9 recovered from the HM-contaminated rhizosphere endowed with multiple activities could play an important role in agricultural practices to augment crop productivity in soils contaminated with HMs. Also, there is an urgent need to control the direct discharge of industrial waste into running water to minimize heavy metal pollution. Furthermore, before the application of HMs in agricultural fields, their appropriate field dosages must be carefully monitored.