ISOLATION OF CADMIUM DEGRADING MICROORGANISMS FROM ELECTROPLATING, STEEL AND BATTERY INDUSTRY

ISOLATION OF CADMIUM DEGRADING MICROORGANISMS FROM ELECTROPLATING, STEEL AND BATTERY INDUSTRY

Sneh Lata ¹, Sukhminderjit Kaur ¹ and Tulika Mishra ^{* 2}

University Institute of Biotechnology ¹, Chandigarh University, Gharuan - 140413, Punjab, India.

Department of Immunology and Microbiology ², Aureus University, Aruba.

ABSTRACT: Increase in industrialization has raised the levels of heavy metal pollution in the country. In order to combat with this problem, many strategies were developed till date, and among them, the use of microorganisms is the most successful technique. The present study was undertaken for isolation, identification, and characterization of heavy metal (Cd) tolerant bacteria from industrial effluents of electroplating, steel and battery industry SAS Nagar, Punjab and Chandigarh (India). The effluents were initially tested for physicochemical parameters (BOD, COD, TDS, TSS, etc.).The cadmium content was quantified through spectrophotometric assay. Initially, a total of 20 bacterial isolates were isolated and screened, then the effect of selected strain on all the parameters of test samples was checked. On the basis of performance to cadmium tolerance, only two isolates were selected. These bacteria could tolerate cadmium up to 1000 ppm and 1500ppm. These isolated bacteria may be helpful for the bioremediation of heavy metal contaminated industrial effluents.

Biochemical oxygen demand, Cadmium, Cadmium tolerance, Heavy metals, pH, Microorganisms

INTRODUCTION: Heavy metals are toxic in nature. Cadmium was discovered by Stromeyer and Hermeny in 1817. A very large amount of cadmium is released into the environment ¹. Cadmium is released into rivers through weathering of rocks, forest fires, etc. Human’s activities are also responsible for the deposition of cadmium by the manufacturing process. So it is entering into the soil and directly into the food chain by absorption through roots. Health hazards to plants, animals, aquatic life, and humans by heavy metals are reported ². Tannery industries are the main sources and causes of such toxic pollutants, such as cadmium ^{3, 4}.

Unprocessed tannery effluents are released into water sources In Bangladesh, which mainly serves as the reason for heavy metal toxicity ^{5, 6}. Many microorganisms have developed a different resistant mechanism for heavy metals ⁷.

Keeping above in view, the present study was designed for the analysis of the physicochemical parameters of the electroplating industry, steel, and battery industry of area SAS Nagar (Punjab, India) and Chandigarh. This study also involves the measurement of cadmium content in these samples by spectrophotometric assay. Also, the strains were isolated from the soil sample of the same area against cadmium, and they were checked for tolerance for different concentrations of cadmium.

MATERIALS AND METHODS:

Sample Collection: Fifteen Soil samples were collected in and near the Industrial Area, Mohali, as well as Chandigarh. Soil and wastewater samples were collected. These samples were placed in sterile plastic bags and bottles, respectively. Bags containing soil samples were first labeled and then placed in the laboratory. Samples were maintained at a temperature of 4 ºC in order to avoid contamination.

Measurement of Physical Parameters:

Color of Soil: Color of soil sample was brown with blackish /grey tinge. It was observed by seeing the color of the soil sample.

pH: To 10 g of air-dried soil 50 ml. of distilled water was added to prepare a suspension (1:5 w/v). The suspension was kept over a shaker for 30 minutes. By using the pH meter, the pH of the soil sample was measured. The pH meter was calibrated using [10] buffers of pH 4.0, 7.0, and 10.0 prior to its use ⁸.

Estimation of Physicochemical Parameters: Various samples (sludge sample, effluent sample) of electroplating Industry, Steel Industry, and battery manufacturing Industries, water sample of various reservoirs has been collected where effluent from the industries are discharged and also the samples from the focal point of various areas. The samples have been tested for BOD, COD, TS, TDS, and TSS ⁹.

Biochemical Oxygen Demand: BOD is defined as the amount of dissolved oxygen needed by the aerobic organisms in order to breakdown the organic material that is present in the water sample at a certain temperature and for a specific period of time. BOD usually determines the degree of organic pollution in water. It is usually measured in milligrams of oxygen consumed per liter of the sample during 5 days of incubation at 20 ºC.

Chemical Oxygen Demand: Chemical oxygen demand (COD) is the measure of oxygen equivalent to the organic content of the sample that is susceptible to oxidation by a strong chemical oxidant. The intrinsic limitation of the test lies in its ability to differentiate between the biologically oxidizable and inert material.

TSS (Total Suspended Solids): Determines the dry weight of particles in a filter. It is a parameter to test the quality of water.  It is measured in mg/L. Filter paper A was weighed. Then 10 ml of water sample was taken, filtered the water sample through weighed filter paper. Filter paper was allowed to dry (in the oven) Again weighed the filter paper B.

TDS (Total Dissolved Solids): It is the measure of a combination of all the organic and inorganic substances present in a liquid (sample) in ionized or granular form. It is a parameter to test the water quality. China Dish was weighed properly. 10ml filtrate (left after filtration). Then that filtrate was transferred to china's dish and boiled the filtrate in china dish until the water was evaporated. Again the china dish was weighed.

Estimation of Cadmium Content in Various Samples: The samples have been tested and quantified for Cadmium through spectrophoto-metric assay ¹⁰. Alizarin red system was used for the preparation of the standard curve of cadmium. Different unknown samples were analyzed for the cadmium concentration by using a standard curve. The absorption spectra of the cadmium-alizarin Red S system in 0 05M H₂SO₄ medium was recorded using the spectrophotometer. The absorption spectrum of the cadmium-alizarin Red S is a symmetric curve with the maximum absorbance coefficient is shown in Fig. 1. In all instances, measurements were made at 422 nm against a reagent blank.

Screening and Isolation of Microorganism for Bioremediation: Primary screening of cadmium resistant bacteria was done to isolate the desired bacteria ¹¹. Sludge samples of various industries, soil samples from banks of water reservoirs, soil samples near the focal points, and effluent samples have been taken and after dilution tested on PDA (Potato dextrose agar medium) and NA (Nutrient agar medium). Various strains have been checked for resistance for cadmium, and pure cultures have been isolated by the standard method of the streak plate method. Out of 20 strains only two were selected for their tolerance of high concentration of cadmium

Effect of Selected Strain on all the Parameters of Test Samples: The strains showing maximum efficiency incubated with various test samples for 24-120 h at various pH. After incubation, the test samples also checked for pH, BOD, COD, TS, TDS, Fluoride, and Chloride.

Heavy Metal Tolerance of Isolated Strains: To assess the ability of isolates to resist Cadmium, isolated microbial strains inoculated on a Nutrient agar/ potato dextrose agar or ionic media (BHB) plates supplemented with different concentration (0.5, 1.0, 2.0, 3.0, 4.0 and 5.0 mM) of cadmium ¹².

RESULTS AND DISCUSSION:

Analysis of Physiological Parameters (KR Aneja 2001): The physiological parameters like BOD, COD TDS, TSS fluoride, and chloride content was analyzed in fifteen samples from different industries, and it is clear from Table 1, 2, 3 that values of all parameters are high. The BOD values ranged from 22 mg/L-47mg/L. The COD values ranged from 230mg/L-540mg/L. Similarly TDS and TSS values ranged from 312mg/L-985 mg/L and 9mg/L-22mg/L respectively. These results are in a very close pattern in various industrial effluent discharges ¹³. The fluoride and chloride content in samples were high. The greater values of all the parameters showed that it might be due to the use of chemicals, i.e., may be organic or inorganic in nature. High values of TDS are due to higher dissolved solids. But the range of TSS values is within the permissible limits that cannot cause damage to agricultural land and aquatic life.

TABLE 1: PHYSICAL PARAMETER FOR SAMPLE OF ELECTROPLATING INDUSTRY

TABLE 2: PHYSICAL PARAMETER FOR SAMPLE OF STEEL INDUSTRY

TABLE 3: PHYSICAL PARAMETER FOR SAMPLE OF BATTERY INDUSTRY

Spectrophotometric Assay of Cadmium: Spectro-photometric assay was done, and a straight line curve was obtained. By using this curve, the concentration of cadmium was determined and found out that the concentration was far above the maximum permissible level, as recommended by FEPA and WHO. The range of concentration of cadmium was 1.02-3.95. It was maximum in sample no. 1 of the battery industry, which shows that the samples were heavily contaminated with toxic cadmium metal. A synthetic reagent CMHBH (cinnamaldehyde-4-hydroxybenzoylhydrazone) was used to find out the toxic metal concentration from wastewater ¹⁴.

FIG. 1: PLOT SHOWING THE SPECTROPHOTOMETRIC ASSAY OF CADMIUM

TABLE 4: CADMIUM CONC. FROM ELECTROPLATING SAMPLES

TABLE 5: CADMIUM CONC. FROM STEEL SAMPLES

TABLE 6: CADMIUM CONC. FROM BATTERY SAMPLES

Isolation of Microorganisms: A total of 20 different strains were isolated from different samples and purified by using nutrient agar media. Out of those, only four were selected on the basis of their resistance to cadmium.

Effect of Selected Strain on all the Parameters of test samples: The selected strains were then tested for all the parameters in order to justify their ability to minimize the cadmium effects, and it is clear from Fig. 2, 3, 4 and 5 that the strains were potent enough to reduce all the parameters i.e., physiological parameters and to make them fall in permissible limits. The readings of all parameters were taken after every 24 h and continued till 120 h. Each and every culture showed a variation in their efficacy and ability.

The novel resistant Bacillus salmalaya strain 139SI was studied ¹⁵ and was potent enough to significantly reduce the chemical oxygen demand. Microbial degradation of paper and pulp industry waste was studied ¹⁶. They utilized fungus for the same purpose.

Cadmium Tolerance: Above isolated strains were tested for different concentrations of cadmium. It is clear from the figures that at concentrations ranging from 100ppm -1500 ppm, the growth of bacterial strain is increasing after every 24 h. E-Cd006 strain showed maximum activity at 1000ppm. Cadmium tolerance by pseudomonas up to 550 µg/ml was measured ¹⁷. Strain steel Cd006 showed good activity against cadmium conc. but the growth decreases after 48 h, especially in 1000 and 1500 ppm. The strain E Cd004 showed its maximum activity at 72 h, but after that, the activity goes decreasing.

CONCLUSION: Heavy metals are hazardous in nature. It is essential to detoxify them. Isolated strains showed a great effect on all physicochemical parameters of samples. Above study intended to isolate such microbial strains having actual efficiency of consuming high levels of cadmium concentrations. Two isolates E-Cd006 and S-Cd005 showed maximum growth in the presence of a high concentration of cadmium. So, these two cultures were selected for further analysis.

ACKNOWLEDGEMENT: The author(s) would like to acknowledge, Chandigarh University, Gharuan, Punjab, India, for providing a research platform and necessary facilities.

CONFLICTS OF INTEREST: All authors have none to declare.

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    How to cite this article:

    Lata S, Kaur S and Mishra T: Isolation of cadmium degrading microorganisms from electroplating, steel and battery industry. Int J Pharm Sci & Res 2020; 11(9): 4403-10. doi: 10.13040/IJPSR.0975-8232.11(9).4403-10.

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