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Author(s): Khushboo Lilaria1, Nisha Gupta2, Jai Shankar Paul*3, Shailesh Kumar Jadhav4

Email(s): 1khushboolilaria@gmail.com, 2nishagupta1210@gmail.com, 3jaishankar_paul@yahoo.com, 4Jadhav9862@gmail.com

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    1School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India
    2School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India
    3School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India
    4School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India
    *Corresponding author: jaishankar_paul@yahoo.com

Published In:   Volume - 2,      Issue - 2,     Year - 2020


Cite this article:
Khushboo Lilaria, Nisha Gupta, Jai Shankar Paul, Shailesh Kumar Jadhav (2020) Comparative Analysis of Oligodynamic Virtue of Various Metals on Bacterial Population. NewBioWorld A Journal of Alumni Association of Biotechnology, 2(2):8-12.

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 NewBioWorld A Journal of Alumni Association of Biotechnology (2020) 2(2):8-12               

ORIGINAL RESEARCH ARTICLE

Comparative Analysis of Oligodynamic Virtue of Various Metals on Bacterial Population

Khushboo Lilaria, Nisha Gupta, Jai Shankar Paul*, Shailesh Kumar Jadhav

School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India.

khushboolilaria@gmail.com, nishagupta1210@gmail.com, jaishankar_paul@yahoo.com, Jadhav9862@gmail.com

*Corresponding Author Email- jaishankar_paul@yahoo.com

ARTICLE INFORMATION

 

ABSTRACT

Article history:

Received

25 June 2020

Received in revised form

14 August 2020

Accepted

18 Sept 2020

Keywords:

Oligodynamic;

bacterial inhibition; copper;

metal;

spread plate

 

Due to increasing pollution, the availability of safe, clean and hygienic drinking water has become very difficult in the recent era. This study was conducted to evaluate the oligodynamic potential of various metals such as copper, bell, silver, aluminium, and brass as well as soil pot against the bacterial colonies in drinking water. The research focuses on the impact of different kinds of metals on bacterial growth in drinking water. The method of research includes the study of the growth and number of bacterial colonies present in drinking water and how much the metals inhibit the growth of bacteria at different time intervals. This study concludes that the copper, bell metal, silver and soil pot efficiently control the growth of bacteria and out of them, copper was found to be most effective in inhibiting bacterial growth. This study helps to provide an easy and cheap method for the purification of drinking water in rural and backward areas.

 


Introduction

Water is the most crucial resource on Earth which is a vital substance for sustaining the entire ecosystem and regulating and maintaining the different physiological functions in a body. Even a single life cannot be imagined without water, as it is vital for the survival and existence of every living organism. According to World Health Organization (WHO) and the United Nations Children Fund (UNICEF), there are around 2.5 billion people who do not have proper sanitation (Bhuiyan et al. 2013). As stated by UNICEF, >3000 children died each day globally due to the consumption of contaminated drinking water (Villapún et al. 2016). Following data obtained from the Central Bureau of Health Intelligence (CBHI) and the Ministry of Health and Family Welfare, waterborne diseases are mainly responsible for most of the infections in children (Barakat 2011; Yatigammana et al. 2011).

There has been an increased risk to human health since the day of the population explosion on Earth and an enhancement in the rate of pollution of water sources. The water gets polluted by several chemicals such as fertilizer and pesticides from agricultural run-off, sewage and food processing waste, heavy metals, chemical waste from industries, urbanization and oil spills.

DOI: 10.52228/NBW-JAAB.2020-2-2-3

The oligodynamic effect was first accredited more than 150 years ago. The word oligodynamic is derived from two Greek words– “Oligos” meaning few and “dynamics” meaning force or power (Klimas et al. 1967). Oligodynamics refers to the ability of heavy metals to exert a lethal effect on bacterial populations.

The ions like mercury, silver, copper, iron, zinc, bismuth, gold and aluminium exert an anti-microbial effect (Alexander et al. 2009; Villapún et al. 2016). However, the utilization of copper as an anti-microbial substance is not a new motive. Copper-based preparations have been used since ancient times to treat wounds, burns, infections, rashes and inflammation. Various mechanisms have been suggested via which copper facilitate antimicrobial activity. The exact mechanism is still not known but some findings suggest that copper damage proteins either by altering the protein conformation or by binding to its active site. Copper may damage nucleic acids by crosslinking within or between the strands of DNA or can damage proteins or can cause the formation of radical ions which can ultimately lead to the destruction of the bacterial cell (Borkow et al. 2009; Villapun et al. 2016). The present study was to undertaken to investigate the oligodynamic potential of various metals including copper, steel, aluminum, brass, and bell metal on bacterial population in different time intervals.

Materials and Methods

All the chemicals used in the current study were of analytical grade and were purchased from HiMedia Laboratories Pvt. Ltd. Mumbai, India, Sigma-Aldrich, USA or Merck India Limited, Mumbai, India.

Preparation of medium

Nutrient Agar Medium (NAM) was used in the current study for the isolation and bacterial growth with a composition (g/L); peptone–5, beef–3, sodium chloride–5 and agar–15g in 1000 mL distilled water. The pH of the medium was kept 7.0±0.2 followed by its sterilization at 15 psi pressure at 121°C for 15–20 minutes. Subsequently, sterile NAM plates were prepared for further use in the study.

Investigating the oligodynamic potential of metals

The metal pots made up of copper, steel, aluminum, brass, and bell metal were selected for the study. The soil pot was also used along with these metals. The above pots were sterilized properly using 70% (v/v) ethanol and were left for drying for about 10-15 min in hot air oven. The metal pots were then filled with 100 mL of tap water and kept at room temperature. After regular interval of time of 12, 24, 36, 48 and 60 h, spread plate method was applied for assessing the bacterial growth and for the analysis of oligodynamic actions of various metal pots via serial dilution.

Isolation of bacteria by spread plate method

For analysis of oligodynamic effect on bacterial growth, stored water samples were taken individually after definite interval of time for isolating the bacteria. Bacterial colony was isolated through spread plate method in sterile NAM plates in triplicates. After the preparation of dilution series from the water sample, 100µL of each water sample of suitable dilution was spread uniformly with the help of a spreader on a sterile NAM plate. The inoculated plates were kept in incubator for 24 h at 37° C.

Morphological characterization of the bacterial isolates

The total numbers of bacterial colonies were counted and the reduction of bacterial density in water sample with increasing holding time interval in the metal pots was recorded. Then the isolated colonies were characterized morphologically.

Results and discussion

Table 1: The number of bacterial colonies in water sample kept in various pots

Hour

Number of bacterial colonies in different metals

Copper

Steel

Aluminum

Brass

Bell

Silver

Soil pot

0

467

Numerous

Numerous

Numerous

312

478

357

12

269

Numerous

Numerous

Numerous

292

344

128

24

147

Numerous

Numerous

Numerous

211

173

111

36

144

Numerous

Numerous

Numerous

187

171

114

48

139

Numerous

Numerous

Numerous

185

172

109

60

138

Numerous

Numerous

Numerous

183

175

114

Microbial analysis of water stored in different storage containers like copper, steel, aluminium, brass, bell metal, and silver was assessed for their oligodynamic potential after a regular interval of time (0-60 hours of incubation). There was a very significant decrease in the bacteria population after 24 hours of incubation in the water sample kept in copper metal, silver and soil pots (Table 1 and 2). However, the water stored for about the same time in brass, aluminium, and steel didn’t show any decrease in the bacterial colonies and hence indicating negligible bacterial population inhibition. Water cultures were tested up to 60 hours of storage and copper metal showed maximum inhibition (about 70.44 %) of bacterial growth (Table 1, 2 and Fig. 1).

The morphological characterization of all the bacterial population of water sample kept in various metal pots was carried out. Majorly, the bacterial population found in the water sample of various metal pots has raised elevation, punctiform with an entire margin and is creamish in colour (Table 3).

The current findings are in line with Shrestha et al. (2010) who reported the oligodynamic virtue of copper metal against the gram-negative bacteria (Salmonella paratyphi). He observed that the bacterial colonies reduced completely within 4 hours of holding time in the copper metal. Packiyam et al. (2016) reported a significant reduction in the bacterial colonies from the water samples kept in mud pots, silver, copper, and brass. Moreover, there was no reduction in the coliform bacteria in the water sample stored in the glass, plastic, ceramics, aluminium, coconut shell and stainless steel. The effect of metals on water-borne and skin-borne pathogens was studied by Dhar (2019) using copper, silver, gold, zinc, and brass metals and reported that silver could eliminate Escherichia coli, Salmonella typhi and Vibrio cholerae by >98% within 24 hours of holding time.

From the current study, it can be concluded that copper metal is the most promising metal in inhibiting the bacterial population within 48–60 hours of holding time. Our results are in corroboration with previous findings. Copper and silver have been shown to have fast reaction rates against the bacterial population according to Varkey (2010) and Praveen (2018).


Figure 1: Bacterial colonies isolated from water sample kept in different pots (a–f) after 60 hours

 

 


Table 2: Percentage inhibition in the bacterial growth by different metallic pot and soil pot

Hour

% inhibition by metals

Copper

Steel

Aluminum

Brass

Bell

Silver

Soil pot

0

0

0

0

0

0

0

0

12

42.39

0

0

0

6.41

28.03

64.14

24

68.52

0

0

0

32.37

63.80

68.90

36

69.16

0

0

0

40.06

64.22

68.06

48

70.23

0

0

0

41.02

64.01

69.46

60

70.44

0

0

0

41.34

63.38

68.06

 

Table 3: Morphological characteristics of bacterial colonies isolated from different metal pots

Metal

Elevation

Margin

Form

Color

Transparency

Copper

Raised

Entire

Punctiform

Cream

Opaque

Steel

Raised

Entire

Punctiform

Cream

Opaque

Aluminum

Raised

Entire

Punctiform

Cream

Opaque

Brass

Raised

Entire

Punctiform

Cream

Opaque

Bell

Raised

Entire

Punctiform

Cream

Opaque

Silver

Flat

Entire

Circular

Cream

Opaque

Soil pot

Raised

Entire

Circular

Cream

Opaque

 


Conclusion

This study was aimed to evaluate the oligodynamic potential of metals including copper, aluminium, steel, brass, bell metal, and silver against the bacterial colonies in the drinking water samples. Among all the tested metal pots, copper showed high depletion (~70%) of microbial colonies after 24–60 hours of incubation. A moderate oligodynamic action was recorded in a water sample kept in a silver and soil pot after 24-60 hours of incubation. Furthermore, the bell metal showed the least (~6-41%) oligodynamic action against the bacterial population. It was observed that the water sample kept in the pot made up of steel, aluminium and brass didn’t show any oligodynamic effect against the bacterial colonies. Copper can be considered to have the best oligodynamic effect on the microbes and thus can be used for the purification of water in a cost-efficient way. Also, soil pots can be used for treating the water in a natural way at a low cost and eco-friendly manner in rural areas.

References

Alexander JW (2009) History of the medical use of silver. Surgical Infections, 10(3): 289–294. https://doi.org.10.1089/sur.2008.9941.

Barakat MA (2011) New trends in removing heavy metals from industrial wastewater. Arabian Journal of Chemistry, 4(4): 361–377.  https://doi.org/10.1016/j.arabjc.2010.07.019.

Bhuiyan A, Mokhtar MB, Toriman ME, Gasim, MB, Ta, GC, Elfithri R, and Razman, M. R. (2013) The environmental risk and water pollution: A review from the river basins around the world. American-Eurasian Journal of Sustainable Agriculture, 7(2):  126–136

Borkow G, Gabbay J (2009) Copper, An Ancient Remedy Returning to Fight Microbial, Fungal and Viral Infections. Current Chemical Biology, 3(3): https://doi.org/10.2174/187231309789054887.

Dhar TP (2019) Oligodynamic Effects Copper Against Water Borne Pathogens. International Journal of Science and Research, 8(4): 08-09. ID ART20196304

Klimas J (1967) Oligodynamic Action of Coins. The American Biology Teacher, 29(9): 750–751.  https://doi.org/10.2307/4441895

Packiyam R, Kananan S, Pachaiyappan S, Narayanan U (2016) Effect of Storage Containers on Coliforms in Household Drinking Water. International Journal of Current Microbiology and Applied Sciences, 5(1): 461–477. https://doi.org/10.20546/ijcmas.2016.501.047

Parveen R, Datta A, Maiti PK (2020) Concentration of capping agent controls size selection, agglomeration and antimicrobial action of silver nanoparticles. Journal of Surface Science and Technology, 36: 137-145. https://doi.org/10.18311/jsst/2020/24875.

Shrestha R, Joshi DR, Gopali J, Piya S (2009) Oligodynamic action of silver, copper and brass on enteric bacteria isolated from water of Kathmandu Valley. Nepal Journal of Science and Technology, 10: 189-193.https://doi.org/10.3126/njst.v10i0.2959.

Varkey AJ (2010) Antibacterial properties of some metals and alloys in combating coliforms in contaminated water. Scientific Research and Essays, 5(24): 3834-3839. https://doi.org/10.20944/preprints201803.0091.v1.

Villapún VM, Dover LG, Cross A, González S (2016) Antibacterial metallic touch surfaces. Multidisciplinary Digital Publishing Institute, 9(9): 1–23. https://doi.org/10.3390/ma9090736

Yatigammana SK, Ileperuma OA, Perera MBU (2011) Water pollution due to a harmful algal bloom: A preliminary study from two drinking water reservoirs in Kandy, Sri Lanka. Journal of the National Science Foundation of Sri Lanka, 39(1): 91–94. https://doi.org/10.4038/jnsfsr.v39i1.2930.

 

 

 



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