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 Table of Contents  
ORIGINAL ARTICLE
Year : 2022  |  Volume : 6  |  Issue : 2  |  Page : 180-184

Screening the in-vitro Antibacterial Activity of Different Naturally Produced Coal and Coal Mediated Commercial Products on Clinically Isolated Pathogens


1 Department of Microbiology, Stamford University Bangladesh, Dhaka, Bangladesh
2 Department of Pharmacy, Stamford University Bangladesh, Dhaka, Bangladesh
3 Department of Pharmacy, Jagannath University, Dhaka, Bangladesh
4 Department of Microbiology, Stamford University Bangladesh, Dhaka, Bangladesh; Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, Shizuoka, Japan

Date of Submission16-Oct-2021
Date of Acceptance16-Nov-2021
Date of Web Publication17-Jun-2022

Correspondence Address:
Mrityunjoy Acharjee
Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, Oya 836, Suruga-ku, Shizuoka, 422-8529

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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/bbrj.bbrj_273_21

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  Abstract 


Background: The rise of microbial illnesses caused by drug-resistant microflora is one of the most serious public health concerns, therefore identifying novel antimicrobial agents is crucial in combating multidrug-resistant bacterial strains. Methods: Present Investigation designated to assess the microbiological status of some naturally produced coal and coal mediated commercial products collected from Barapukuria Coal Mine Dinajpur and different retailer in Dhaka city, respectively through conventional cultural and biochemical tests. Moreover, the anti-bacterial potential of both categories of coal samples was also aimed to be checked by the agar well diffusion method and minimal inhibitory concentration (MIC) assay. Results: Out of 10 samples, total viable bacteria was estimated at high range in all the samples excluding the charcoal. In case of specific bacteria, only Staphylococcus spp. was found in both types of samples while Klebsiella spp. was found in coal mediated commercial products. Both categories of samples showed their anti-bacterial activity with satisfactory range of zone diameter (9 mm–26 mm). However, the naturally produced coal was unable to show their anti-bacterial activity against Bacillus spp., Shigella spp. and Vibrio spp. through agar well diffusion methods. Meanwhile, the minimal concentration of all the samples was successfully quantified as MIC value at which the bacterial growth was retarded. In case of naturally produced coal, the lowest concentration 4 mg/ml was recorded for wooden coal powder against Escherichia coli while the maximum concentration was estimated at 34 mg/ml. Conversely, the MIC value was varied between 4 mg/ml to 24 mg/ml in case of coal mediated commercial products. Conclusion: The results of this study suggest that the naturally produced coal and coal-mediated products have excellent antibacterial characteristics which indicating their economic potential for treating various infectious disorders.

Keywords: Anti-bacterial activity, coal mediated commercial products, coal samples, MIC


How to cite this article:
Shreya SS, Islam T, Ishma T, Polin IS, Nazia FM, Acharjee M. Screening the in-vitro Antibacterial Activity of Different Naturally Produced Coal and Coal Mediated Commercial Products on Clinically Isolated Pathogens. Biomed Biotechnol Res J 2022;6:180-4

How to cite this URL:
Shreya SS, Islam T, Ishma T, Polin IS, Nazia FM, Acharjee M. Screening the in-vitro Antibacterial Activity of Different Naturally Produced Coal and Coal Mediated Commercial Products on Clinically Isolated Pathogens. Biomed Biotechnol Res J [serial online] 2022 [cited 2022 Jul 7];6:180-4. Available from: https://www.bmbtrj.org/text.asp?2022/6/2/180/347712




  Introduction Top


Antibiotic misuse and mass manufacture have resulted in a worldwide problem with antibacterial infection therapy. As a result, any opportunity to reduce the number of antibacterial drugs used will help to reduce the spread of pathogenic bacterial resistance. Activated charcoal was once thought to be the universal antidote in the last few decades. Nowadays it continues to be promoted as a potent for natural treatment. Normally activated charcoal is derived from common charcoal.[1] Manufactures make it activated by heating the common charcoal in the presence of gas.[2] For the past 20 years, activated charcoal has been frequently utilized to treat poisoned people.[3] Activated charcoal has numerous applications, one of which is the use of activated charcoal in medicine. According to World Health Organization, activated charcoal is an essential medicines which is the safest and most effective remedies needed in a health system.[4] In various countries, different types of activated charcoal tablets or capsules are used to treat diarrhea, indigestion, flatulence, poisoning, overdoses of oral consumption, and other ailments.[5] A review article published in the “journal of pediatrics” in 1963 is attributed with encouraging the use of charcoal in poisoned patients to become more widespread.[6]

Generally, carbonaceous source materials such as bamboo, coconut, husk, willow peat, wood, coir, lignite, coal, and petroleum pitch are used to make activated coal.[7] After obtaining the charcoal, it is broken down into fine granular form. Further processes, such as treating the charcoal with water, O2, CO2, specific acids, and other chemicals, are required to activate it. This activating process removes impurities and creates, fine small granules. Gas adsorption determines that one gram of activated Carbon has a surface area in access of 3000 m2 (3200 sq ft) due to its high degree of microporing.[8] It is declared that activated charcoal will absorb most drugs and toxin rather than the normal charcoals.[9],[10]

Activated charcoal is an allowed substance used by organic farmers in both livestock production and wine making. In case of livestock production it is used as pesticide, animal feed additive, processing aid, nonagricultural ingredient and disinfectants.[11] In organic wine making purposes activated carbon is used as a processing agent to adsorb brown color pigments from white grape concentrates.[12] Sometimes it is used as biochar.

Due to its multifunctional nature, activated carbon filtration is an effective water treatment process. Depending on the type of contamination, several types of activated carbon filtering processes and equipment are recommended.[12]

Considering all these facts this study was designed to investigate the antimicrobial traits of different coal samples (naturally produced and coal mediated commercial products) available in Bangladesh through the agar well diffusion method and the micro dilutions method.


  Materials and Methods Top


Study area, sampling, sample processing and microbiological analysis

Total of 10 samples (5 categories of naturally produced coal such as Wooden coal powder, Coal Ash, Solid coal powder, Charcoal, Coal dust from Barapukuria coal mine and coal mediated commercial products coal such as Charcoal face scrub, Charcoal face masks, Charcoal Ponds face wash, Charcoal Men's face wash, Charcoal Pepsodent from different shop) were randomly collected during July 2019 to September 2019 following standard protocol (APHA 1998). Samples were transported to the laboratory to conduct both microbiological assay and in-vitro antimicrobial assay.[13],[14]

Microbiological analysis

Enumeration of total viable bacteria and fungi

For the enumeration of total viable bacteria (TVB) and total fungal load, 0.1 ml of each sample from dilution 10-2 was spread into Nutrient Agar and Sabouraud Dextrose agar media by spread plate technique. Then plates were incubated at 37°C for 24 h for TVB count and 25°C for 48 h for total fungal count.[15],[16]

Enumeration of Escherichia coli and Klebsiella spp

For the detection of Klebsiella spp. as coliform 0.1 ml of each sample from dilution 10-2 was spread into MacConKey agar by spread plate technique and then incubated at 37°C for 24 h. For the isolation of E. coli, 0.1 ml of suspension from the dilution 10-2 were spread over on Eosine-Methylene Blue (EMB) agar for each samples and incubated at 37°C for 24 h. Presence of E. coli was further confirmed by the appearance of bluish-black colonies with green metallic sheen on the EMB agar as well as by biochemical tests.[17]

Estimation of Staphylococcus spp. and Pseudomonas spp

For the detection of Staphylococcus spp. and Pseudomonas spp. 0.1 ml of each sample from dilution 10-2 was spread into Mannitol Salt agar and Pseudomonas agar by spread plate technique and incubated at 37°C for 24 h.[18]

Assay of antimicrobial activity of charcoal against the clinical isolates using agar well diffusion method

The Muller Hinton Agar (MHA) were prepared followed by modified agar well diffusion method. Fresh culture suspension of microorganisms (previously isolated from clinical samples) including E. coil, Klebsiella spp., Staphylococcus spp., Bacillus spp., Pseudomonas spp., Shigella spp and Vibrio spp. from the previous stock media was lawned on the respective plates. Over the agar plates, wells were punched by using sterile Borer. About 100 μl of each sample were added to the wells with negative and positive controls.[13] Plates were incubated for 24 h at 37°C. After incubation, the diameter of inhibitory zones formed around each wells were measured in mm.

Determination of minimal inhibitory concentration (MIC)

The MIC of the coal samples against the bacterial species were performed in Mueller Hinton (MHA) broth through microdillution methods.[19],[20] 100 μl of bacterial suspention (turbidity adjusted with 0.5 McFarland standard) was inoculated into the sterile vials those were previously filled with Mueller Hinton (MH) broth (Oxoid Ltd, England). Then different concentration of coal and coal mediated products were subjected to introduced into the inoculam containing broth. After 37°C for 24 h of incubation the smallest concentration of samples which could retard the multiplication of the tested bacteria as indicated by measuring zero the zero density, was regarded as MIC.


  Results Top


All the samples of both categories were found to be contaminated with heterotrophic bacteria in high amount [Table 1]. The result of bacterial contamination in the coal samples was expressed by three different way like high growth rate (+++), moderate growth rate (++) and low growth rate (+).
Table 1: Microbiological status of different coal and coal mediated products

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From naturally produced coal category, the Wooden coal powder, Solid coal powder and Coal dust samples showed high growth of TVB, Coal Ash exhibited moderate growth and there was no growth present in Charcoal samples. Likewise, Charcoal face scrub, Charcoal face masks, Charcoal Ponds face wash, and Charcoal Pepsodent were found to be highly contaminated by total heterotrophic bacteria while the moderate growth of total heterotrophic bacteria was present in Charcoal Men's face wash [Table 1]. In case of specific microflora, Staphylococcus spp. was noticed in 10 samples of both categories. Among the 10 samples, the low growth of Staphylococcus spp. was quantified for coal and Charcoal Men's face wash while the Klebsiella spp. was found only in the Charcoal face scrub, Charcoal facemasks, Charcoal Ponds face wash, and Charcoal Pepsodent within moderate range [Table 1].

In vitro anti-bacterial activity of different coal samples

Many studies on the antibacterial action of various natural and commercial items, such as herbs, leaves, fruits, tree routes, oil, cosmetics, and preservatives, have previously been published.[21],[22],[23],[24] This study was effective in determining the antibacterial activity of various naturally formed coal and commercially produced coal samples, which are uncommon not only in our nation but also globally. Most of the samples showed antibacterial activity against the tested bacteria [Table 2]. Wooden coal powder was able to produced 19 mm zone of inhibition against E. coli, Klebsiella spp., and Staphylococcus spp. and 20 mm against Pseudomonas spp., coal ash sample produced zone diameter 24 mm against E. coli, 18 mm against Klebsiella spp., 23 mm against Staphylococcus spp. and 20 mm against Pseudomonas spp., solid coal powder sample created zone 20 mm against Klebseilla spp. and Pseudomans spp. while 26 mm and 16 mm zone against E. coli and Staphylococcus spp. respectively. Charcoal showed zone diameter 19 mm against E. coli and Staphylococcus spp. while 22 mm and 24 mm were recorded against Klebsiella spp. and Pseudomonas spp. respectively.
Table 2: Antimicrobial activity of different coal and coal mediated products

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Finally, coal dust displayed 20 mm zone of inhibition against E. coli and Klebsiella spp., 22 mm against Staphylococcus spp. and Pseudomonas spp. [Table 2]. All the samples were unable to produce their anti-bacterial activity against Bacillus spp., Shigella spp. and Vibrio spp.

Coal Mediated Commercial Products exhibited their antibacterial activity against all the bacteria tested in this study [Table 2]. Charcoal face scurb showed their antibacterial activity against all the tested bacteria, the massive zone diameter was quantified 19 mm against Bacillus spp. and the lowest zone was 9 mm against Vibrio spp. The maximum zone length 18 mm was produced by Charcoal face masks against Shigella spp. and minimum zone was 10 mm against Vibrio spp. while the highest zone of inhibition was estimated 21 mm for Charcoal Ponds face wash against E. coli, Klebseilla spp and Bacillus spp. and 15 mm against Vibrio spp. Likewise, Charcoal Men's face wash was able to create maximum zone diameter 23 mm against Bacillus spp. while the Charcoal Pepsodent showed 24 mm zone diameter against Staphylococcus spp.[Table 2].

Detection of MIC of the samples

In addition, the in vitro anti-bacterial activity of the samples was further supported by observing the result of MIC [Table 3]. In this parameter all the samples were found to show their anti-bacterial activity against different tested bacteria; i.e., E. coli, Klebsiella spp., Pseudomonas spp., Bacillus spp., Shigella spp., Staphylococcus spp., and Vibrio spp. In case of agar well diffusion methods, the naturally produced coal samples were not capable to show their antibacterial activity against Bacillus spp., Shigella spp. and Vibrio spp. due to the insufficient concentration of the samples but the appropriate concentration of the samples were evaluated as MIC [Table 3]. The highest MIC was scored at 34 mg/ml against Bacillus spp. particularly for the five categories of naturally produced coal samples while the lowest value was noted to be 4 mg/ml especially for Wooden coal powder [Table 3]. On the other hand, the maximum MIC was quantified at 24 μL against Bacillus spp. for the sample Charcoal face masks and Charcoal Ponds face wash while the most effective sample was Charcoal Pepsodent which revealed the MIC 4 mg/ml to stall the growth of Staphylococcus spp. However, the most common MIC value was recorded within range of 10–17 mg/ml for Commercially Coal samples [Table 3].
Table 3: Minimal inhibitory concentration of different coal and coal mediated products

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  Discussion Top


In the 1700s, the adsorbent characteristics of charcoal were documented, and the first clinical applications were made in the early 1800s. A variety of carbon-containing materials can be used to make activated charcoal including low-ash wood pulp, coal, lignite, and rye starch. After chemically obtaining charcoal (almost pure carbon), it is broken down into a fine granular state. It is then treated with steam, oxygen, carbon dioxide, specific acids, and other chemicals in order to activate it.[2],[3],[4],[5],[6]

The adsorbing properties and potential clinical effects of charcoal have been reported in several research. Despite this, until the last 20 years, charcoal was not commonly acknowledged as an important aid in the treatment of poisoned patients.[9]

Several studies have previously described the beneficial advantages of bamboo charcoal, which is currently in high demand in many industrialized countries such as South Asia, India, and China.[25],[26] The reports on the anti bacterial activity of different types of naturally produced coal samples as well as Coal Mediated Commercial Products have not yet published many more in the past. However, our study findings are quite similar with another research group who found the antibacterial effects of charcoal against E. coli, Staphylococcus aureus. Pseudomonas aerugenosa and Pseudomonas putida.[27]

From the outcomes of our study, most of the coal mediated commercial product revealed satisfactory range of zone diameter against all the tested bacteria beside the naturally produced charcoal. Several air borne diseases are available those can create skin diseases. To prevent such skin diseases the charcoal face scrub, charcoal face maskes, charcoal Ponds face wash, charcoal Men's face wash and charcoal pepsodent play important role not only as the protector of the portal of entry of the diseases causing agents but also those can absorb the toxin substances in the body produced by bacteria and fungi.[4]

Meanwhile, the coal samples also have some negative impact on the human health especially for the coal workers. Several diseases like pneumoconiosis and anthracosis can be transpired due to the inhalation of excessive coal dust. Anthracosis is a chronic pulmonary condition characterized by black pigmentation of the bronchial mucosa, which is caused by carbon buildup in the lungs and is sometimes linked to pulmonary tuberculosis.[28] Therefore, expanded research work is urgently required to evaluate the beneficial effects of such natural resources as well as to sort out the appropriate way to reduce the negative impact of the substances.

However, as there are no reports on the antimicrobial activity of coal and coal-mediated products have been published yet, this comprehensive study of the microbiological and antimicrobial activity of the available coal and coal-mediated products in Bangladesh could pave the way for future research on this product for better results.


  Conclusion Top


The antibacterial effectiveness of naturally formed coal samples and coal medicated products against clinical pathogens such as E. coli, Pseudomonas spp., Vibrio spp., Shigella spp., Klebsiella spp., Staphylococcus spp. and Bacillus spp. has now been experimentally proven. Along with the naturally produced coal samples, there are numerous coal mediated products available in the market therefore it was necessary to evaluate their microbiological condition as well as their antibacterial activity. Apart form the antibacterial properties of coal samples, it is also very much useful in terms of water purification materials in almost all over the world. Our pilot scale experimental data would be evocative for researcher to conduct more details investigation on this theme more elaborately to know the specific compound that govern the antibacterial activity and purification needs to be demonstrated.

Acknowledgement

We thank Stamford University Bangladesh for providing financial and technical support.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Tables

  [Table 1], [Table 2], [Table 3]



 

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