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Original Article
6 (
1
); 3-8
doi:
10.25259/JISH_51_2022

In vitro antimicrobial activity of nine homoeopathic preparations in different volumes against Staphylococcus aureus and Escherichia coli

Department of Pathology and Microbiology, National Institute of Homoeopathy, Kolkata, West Bengal, India
Postgraduate trainee, National Institute of Homoeopathy, Kolkata, West Bengal, India
Intern, National Institute of Homoeopathy, Kolkata, West Bengal, India
Corresponding author: Dr. KumaraVel V., Department of Pathology and Microbiology, National Institute of Homoeopathy, Kolkata, West Bengal, India. drkumaravelnih@gmail.com
Licence
This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-Share Alike 4.0 License, which allows others to remix, transform, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

How to cite this article: Kumaravel V, Mondal M, Aggarwal M, Shaw N, Singh S, Bhattacharya S, et al. In vitro antimicrobial activity of nine homoeopathic preparations in different volumes against Staphylococcus aureus and Escherichia coli. J Intgr Stand Homoeopathy 2023;6:3-8.

Abstract

Objective:

The present study aims to evaluate the efficacy of antimicrobial activities of nine homoeopathic preparations through an in vitro study.

Materials and Methods:

The antimicrobial activity of nine homoeopathic preparations was evaluated through broth microdilution according to clinical and laboratory standards institute with slight modifications (M07-A10).

Result:

The mean and standard deviation of homoeopathic preparations against S. aureus and E. coli show lesser values than control and ethyl alcohol at lower volumes (10 μL and 15 μL, respectively). The broth microdilution assay provides the information that seven homoeopathic preparations exhibited significant results against S. aureus at 10 μL and eight homoeopathic preparations exhibited significant results against E. coli at 15 μL.

Conclusion:

The ultra-diluted homoeopathic preparations at different volumes especially lower volumes showed significant results against S. aureus (10 μL) and E. coli (15 μL) when compared to the negative control (91% ethyl alcohol). Further, in vivo studies may be carried out to confirm their mode and mechanism of action of homoeopathic preparations.

Keywords

Homoeopathy
Antimicrobial activity
Staphylococcus aureus
Escherichia coli
Broth microdilution method

INTRODUCTION

The global incidence and mortality rate of anti-microbial resistance (AMR) in 2019 was 1.27 million and 5 million, respectively.[1] Staphylococcus aureus and Escherichia coli are among the six pathogenic microbes that are responsible for AMR in India. The staphylococcus genus contains approximately 30 species of medical importance, mainly S. aureus which is responsible for hospital-acquired infections.[2] It produces pyogenic infections in the skin and various other organs.[3]

E. coli belongs to a group of diarrheagenic Enterobacteria.[3] Enterobacteria contain five species of medical importance, mainly E. coli, which are normally present in the human intestinal flora. The virulent strain of E. coli produces urinary bladder infections, wound infections and diarrhoea, among other disorders. E. coli is most commonly isolated among the Enterobacteria and is also the main isolate from the urine samples.[2]

Homoeopathy is one of the major alternative medicine systems used worldwide. Homoeopathic medicines are administered in ultra-diluted forms; they are effective in various disorders, which can be confirmed through clinical trials. However, there is a lack of in vitro and in vivo studies in homoeopathy.

Denny, in an in vitro study against various cell lines, observed that antiproliferative properties of polyketides can be isolated from Myristica sebifera leaves.[4] A study by Sultana suggested that the methanolic extract of Atropa belladonna showed a significant degree of antimicrobial activity against medically important bacteria like S. aureus, E. coli and Klebsiella pneumonia.[5] Munir et al., in another study, also observed that the ethanolic extract of A. belladonna showed more significant anti-microbial activity against S. aureus than E. coli.[6] A study by Salari et al. suggests the antibacterial activity of Eucalyptus globulus leaf extract was determined for different isolates of S. aureus. The results suggest the possible therapeutic role of E. globulus leaf extract in the treatment of respiratory tract infection.[7] A study was carried out at Maranhão State University and Paraíba Federal University to evaluate the antimicrobial activity of Ruta graveolens. Bacillus cereus and S. aureus were the two bacteria that were most responsive to the antibacterial activity of Ruta R. graveolens against Gram-positive and Gram-negative bacteria with inhibition zones.[8]

Several homoeopathic remedies are indicated in cases of infections and inflammations. However, in homoeopathy, there is no specific remedy for specific infections. Therefore, homoeopathy is limited to symptomatology and Materia Medica descriptions. The selection of nine homoeopathic preparations is based on source books on homoeopathy and other traditional systems.

M. sebifera is indicated in inflammation of the skin, cellular tissue and periosteum and also in traumatic infections, parotitis and carbuncles. It has an almost specific action in panaritium.[9] A. belladonna is indicated in erysipelatous inflammation with swelling, phlegmonous erysipelas, bright red and radiating. Vesicular erysipelas (when fever is violent). Intense erysipelatous fever, accompanied by inflamed swellings, passing even into gangrene.[10] E. globulus is indicated for chronic desquamative nephritis and pyelonephritis.[11] It is a powerful antiseptic and destructive to a low form of life. It is also useful in treating foul and indolent ulcers with glandular enlargement and nodular swellings.[9] R. graveolens is a remedy for periostitis and erysipelas following bruises and mechanical injuries.[12]

We aimed to evaluate the efficacy of antimicrobial activities of these nine homoeopathic preparations against S. aureus and E. coli through an in vitro study.

MATERIAL AND METHODS

Media and chemicals

All media were purchased from HiMedia, Mumbai, Maharashtra, India. All chemicals and reagents were purchased from MERCK company, Kolkata, West Bengal, India.

Homoeopathic preparations

Nine homoeopathic preparations: M. sebifera 12C, M. sebifera 30C, A. belladonna Q, E. globulus 3C, E. globulus 12C, E. globulus 30C, R. graveolens 12C, R. graveolens 30C, and R. graveolens 200C were purchased from Hahnemann Publishing Co. Private Ltd, Kolkata, West Bengal, India.

Controls

  1. Positive control (gentamycin): As per clinical and laboratory standards institute (CLSI) guidelines

  2. Negative control (91% ethyl alcohol): A solvent used for homoeopathic preparations

  3. Control: Untreated group.

Microorganisms

S. aureus (IDH-7473) and E. coli (IDH-13003) isolates were obtained from the National Institute of Cholera and Enteric Diseases, ICMR, Kolkata 700010. As shown in Figure 1, the cultural characteristics of E. coli include large, white, round and moist colonies.

Figure 1:
Cultural characteristic of E. coli. Two nutrient agar plates for each microbe were prepared in a petri dish and media was incubated for 24 h at 37°C. The characteristics of E. coli shows large colonies (>2 mm), white, round and moist colonies. E. coli: Escherichia coli.

Preparation of microbial culture

Cultures of S. aureus and E. coli were placed in Petri plates containing nutrient agar (Hi-Media Catalogue No: M001) and the microbes were opened aseptically in a laminar airflow. Two nutrient agar plates for each microbe were prepared in a petri dish for identification and confirmation of its colony. The media were incubated for 24 h at 37°C.

Preparation of inoculum

The microbes were picked up with a sterile loop from respective culture plates in sterile saline and cell concentration was adjusted to final concentrations of 0.5 McFarland’s standard (1.5 × 105 CFU/mL).[13-15]

Broth microdilution technique

The broth-microdilution assay was performed to identify the lowest volumes of homoeopathic preparation that could inhibit the microbes. The antimicrobial activities of nine homoeopathic preparations in different volumes were performed in a 96-well plate (Elisa plate flat bottom, HSN Code- 39233090, Maxome Labsciences Pvt Ltd.) having a total capacity of 400 μL volume with a flat bottom as described in CLSI with slight modifications (M07-A10).[13-15] First, different volumes of homoeopathic preparations 5–30 μL (selection of volume is based on the pre-experiment assay) were used instead of serial dilution of antimicrobial agents. Second, gentamycin (Positive Control) and 91% ethyl alcohol (Negative Control) were used to identify the drug action. Total volume of 200 μL was used out of 400 μL. MH broth (185 μL) and bacterial suspensions (10 μL) were added to a 96-well microliter plate except for the 12th column. The nine homoeopathic preparations (5 μL) were added up to the 9th column. The gentamycin (5 μL) was added to the 10th column as positive control and 91% of ethyl alcohol (5 μL) was added to the 11th column as a negative control. The 12th column contained 190 μL of MH broth and bacterial suspensions (10 μL) as control. All test tubes were incubated at 37°C for 24 h. The test was repeated with the same procedure for other five different volumes of homoeopathic preparations (10 μL, 15 μL, 20 μL, 25 μL and 30 μL) after withdrawing the same amounts of media from 96-well plates before adding bacterial suspensions (10 μL). All assays were performed in triplicate for accuracy and reproducibility. The 96 microplates were examined using a visible spectrophotometer (Automated ELISA Microstrip Analyser, Model: ELAN 30s) at an absorbance of 630 nm to find the optical density (OD)/ turbidity for identification of the inhibitory properties of homoeopathic preparations. The graphical abstract [Figure 2] depicts the key findings of the study.

Figure 2:
Graphical abstract.

Statistical analysis

The data were analysed by applying one-way analysis of variance to know a significant difference (P < 0.05 was considered significant) among the groups followed by Dunnett’s post hoc test (only the groups that showed significant results in one-way analysis of variance) to identify significance among groups (Medicines, Ethyl alcohol and Control – P < 0.016 was considered significant). Ethical approval was not required for this study.

RESULTS

The mean and standard deviation of control were observed as 1.334 ± 0.25 against S. aureus and 1.038 ± 0.22 against E. coli [Tables 1 and 2] respectively.

Table 1: Mean and standard deviation of different concentrations of nine homoeopathic potencies in 96-well plates along with gentamycin (Positive Control), 91% alcohol (Negative Control) and only control (broth and Microbe) against Staphylococcus aureus.
Drug 5 uL 10 uL 15 uL 20 uL 25 uL 30 uL
M. sebifera 12C 1.06±0.19 0.85±0.26 0.96±0.20 0.39±0.03 2.04± 0.26±0.08
M. sebifera 30C 0.26±0.29 0.87±0.16 0.93±0.25 0.30±0.04 2.1±0.16 0.26±0.06
Belladonna Q 1.13±0.36 1.66±0.12 1.73±0.63 0.82±0.21 2.54±0.22 0.55±0.06
E. globulus 3C 1.11±0.20 0.77±0.15* 1.04±0.16 0.28±0.08 2.24±0.19 0.21±0.04
E. globulus 30C 1.15±0.18 0.66±0.10* 0.92±0.37 0.32±0.04 2.17±0.28 0.21±0.09
E. globulus 12C 1.23±0.13 0.74±0.19* 1.01±0.38 0.31±0.06 2.17±0.22 0.19±0.03
R. graveolens 12C 1.51±0.17 0.62±0.15* 0.96±0.26 0.32±0.02 2.32±0.28 0.24±0.04
R. graveolens 30C 1.39±0.15 0.83±0.27* 1.45±0.52 0.31±0.04 2.44±0.31 0.26±0.09
R. graveolens 200C 1.36±0.36 0.66±0.28* 1.02±0.35 0.32±0.06 1.93±0.77 0.28±0.05
Positive control 0.03±0.008 0.05±0.03 0.03±0.02 0.12±0.17 0.12±0.16 0.08±0.02
Negative control 1.29±0.23 0.84±0.34 0.81±0.47 0.32±0.05 2.23±0.19 0.34±0.13
Control 1.334±0.25

M. sebifera: Myristica sebifera, E. globulus: Eucalyptus globulus, R. graveolens: Ruta graveolens, *: indicates significant P values

Table 2: Mean and standard deviation of different concentrations of nine homoeopathic potencies in 96-well plates along with gentamycin (Positive Control), 91% Alcohol (Negative Control) and only control (broth and Microbe) against Escherichia coli.
Drug 5 uL 10 uL 15 uL 20 uL 25 uL 30 uL
M. sebifera 12C 0.98±0.149 0.971±0.15 0.52±0.42 0.29±0.22 0.39±0.14 0.15±0.02
M. sebifera 30C 1.12±0.197 1.14±0.22 0.71±0.19* 0.65±0.16 0.37±0.05 0.10±0.06
Belladonna Q 1.25±0.27 1.35±0.15 0.62±0.04* 1.09±0.21 0.45±0.02 0.09±0.02
E. globulus 3C 1.034±0.28 1.07±0.20 0.73±0.24* 0.62±0.19 0.27±0.01 0.11±0.03
E. globulus 30C 1.17±0.40 1.21±0.20 0.54±0.24* 0.80±0.06 0.31±0.06 0.08±0.02
E. globulus 12C 1.04±0.15 1.27±0.17 0.67±0.13* 0.69±0.13 0.30±0.02 0.09±0.009
R. graveolens 12C 1.03±0.21 1.20±0.15 0.55±0.11* 0.80±0.17 0.29±0.05 0.06±0.007
R. graveolens 30C 1.15±0.26 1.13±0.28 0.47±0.29* 0.69±0.08 0.30±0.04 0.04±0.02
R. graveolens 200C 1.25±0.25 1.31±0.30 0.57±0.34* 0.73±0.10 0.33±0.01 0.10±0.008
Positive control 0.009±0.01 0.03±0.008 0.04±0.006 0.04±0.03 0.12±0.073 0.050±0.01
Negative control 0.86±0.20 1.01±0.13 0.81±0.31 0.638±0.27 0.348±0.05 0.056±0.01
Control 1.038±0.22

M. sebifera: Myristica sebifera, E. globulus: Eucalyptus globulus, R. graveolens: Ruta graveolens, *: indicates significant P values

M. sebifera 12C in 10 μL, 15 μL and 30 μL has shown statistically significant results against S. aureus [Table 3] and in 15 μL, 20 μL against E. coli [Table 4]. M. sebifera 30C in 15 μL has shown statistically significant results against S. aureus [Table 3] and in 25 μL against E. coli [Table 4].

Table 3: One-way ANOVA analysis and Dunnett’s post hoc test of different concentrations of nine homoeopathic potencies against Staphylococcus aureus.
Drug 5 uL 10 uL 15 uL 20 uL 25 uL 30 uL
ANOVA P<0.05 Post hoc test P<0.016 ANOVA P<0.05 Post hoc test P<0.016 ANOVA P<0.05 Post hoc test P<0.016 ANOVA P<0.05 Post hoc test P<0.016 ANOVA P<0.05 Post hoc test P<0.016 ANOVA P<0.05 Post hoc test P<0.016
MS12C 0.067232 - 0.0066388* 0.002315481* 0.0079288* 0.006043* 0.000000000035* 0.000000051 0.000358* 0.001976144 0.000000000234* 0.000000017*
MS 30C 0.28923 - 0.001528* 0.000593061* 0.015244* 0.00609428* 0.0000000000028* 0.000000019* 0.000000017* 0.000001557 0.000000000022* 0.000000015*
Bell Q 0.947744 - 0.000010185852* 0.005525933* 0.003785* 0.122607 0.000000005337* 0.000610009 0.000000001469* 0.000000081* 0.000000000268* 0.000000726
EG 3C 0.155185 - 0.000562* 0.00011* 0.014426* 0.014698* 0.000000000005* 0.000000024* 0.000000016543* 0.000001148* 0.000000000009* 0.000000006*
EG 30C 0.262408 - 0.000084758129* 0.000006979* 0.028344* 0.02192326 0.000000000003* 0.000000022* 0.000000354035* 0.000024841 0.000000000020* 0.000000012*
EG 12C 0.652866 - 0.000538* 0.0001263* 0.038543* 0.067281 0.000000000004* 0.000000025* 0.000000078564* 0.000006757 0.000000000007* 0.000000005*
RG 12C 0.144896 - 0.000069579155* 0.000008191* 0.017967* 0.010689* 0.000000000003* 0.000000021* 0.000000070919* 0.000003264 0.000000000013* 0.000000009*
RG 30C 0.691525 - 0.002885* 0.00180796* 0.000466* 0.580944033 0.000000000003* 0.000000022* 0.000000061141* 0.000002277 0.000000000034* 0.000000021*
RG 200C 0.850499 - 0.000466* 0.000172* 0.033775* 0.060246* 0.000000000005* 0.000000029 0.004514* 0.058122377* 0.000000000023* 0.000000017*
NC - - - 0.005325109 0.015604 0.000000027 0.000001444 0.000000127

ANOVA: Analysis of variance. The OD was analysed by one-way ANOVA Analysis to know the significant difference (P<0.05 was considered as significant.) among the groups. The Dunnett’s post hoc test (only those groups show a significant result in one-way analysis of variance) to identify significance between groups (Medicines, alcohol and control). P<0.016 was considered as significant. MS: Myristica sebifera, Bell: Belladonna, EG: Eucalyptus globulus, RG: Ruta graveolens, NC: Negative control, *: indicates significant P values

Table 4: One-way ANOVA analysis and Dunnett’s post hoc test of different concentrations of nine homoeopathic potencies against Escherichia coli.
Drug 5 uL 10 uL 15 uL 20 uL 25 uL 30 uL
ANOVA Post hoc test ANOVA Post hoc test ANOVA Post hoc test ANOVA Post hoc test ANOVA Post hoc test ANOVA Post hoc test
MS12C 0.21857 0.748144 0.021125* 0.010175* 0.000024296811* 0.000014093* 0.000000016223* 0.000008451 0.000000000001* 0.000000029
MS 30C 0.065327 0.373273 0.049621* 0.009359* 0.002667* 0.607293 0.000000001215* 0.000001241 0.000000000001* 0.000000024
Bell Q 0.014099* 0.117223* 0.001094* 0.005362* 0.005107* 0.000168* 0.001816* 0.001459* 0.000000002119* 0.000004274 0.000000000001* 0.000000012
EG 3C 0.277474 0.78011 0.076431 0.002415* 0.014658 0.000000000218* 0.000000186* 0.000000000001* 0.000000018
EG 30C 0.133669 0.095407 0.00476* 0.000889* 0.003686* 0.014658 0.000000000614* 0.000000483* 0.000000000001* 0.000000012
EG 12C 0.134044 0.015248* 0.033768 0.01792* 0.00156* 0.002813* 0.002224* 0.000000000332* 0.000000296* 0.000000000001* 0.000000012
RG 12C 0.207793 0.084433 0.002006* 0.000108* 0.007874* 0.040108 0.000000000418* 0.000000331* 0.000000000076* 0.000000008
RG 30C 0.064337 0.373273 0.002404* 0.000725* 0.002069* 0.001294* 0.000000000414* 0.000000352* 0.00000000000037* 0.000000006385
RG 200C 0.009025 0.088694* 0.001094* 0.055933* 0.018842* 0.006683* 0.003171* 0.00378* 0.000000000467* 0.000000463* 0.000000000001* 0.000000014
NC 0.12948 0.77256 0.110903 0.00661 0.000000846 0.000000008

ANOVA: Analysis of variance. The OD was analysed by one-way ANOVA Analysis to know the significant difference (P<0.05 was considered as significant.) among the groups. The Dunnett’s post hoc test (only those groups show a significant result in one-way analysis of variance) to identify significance between groups (Medicines, alcohol and control). P<0.016 was considered as significant. MS: Myristica sebifera, Bell: Belladonna, EG: Eucalyptus globulus, RG: Ruta graveolens, NC: Negative control, *: indicates significant P values

A. belladonna Q in 25 μL has shown statistically significant results against S. aureus [Table 3] and in 5 μL, 10 μL, 15 μL and 20 μL against E. coli [Table 4].

E. globulus 3C in 10 μL, 15 μL, 20 μL, 25 μL and 30 μL has shown statistically significant results against S. aureus [Table 3] and in 25 μL against E. coli [Table 4]. E. globulus 12C in 10 μL, 20 μL and 30 μL has shown statistically significant results against S. aureus [Table 3] and in 10 μL, 15 μL, 20 μL and 25 μL against E. coli [Table 4]. E. globulus 30C in 15 μL and 25 μL has shown statistically significant results against E. coli [Table 4].

R. graveolens 12C in 10 μL, 15 μL, 20 μL and 30 μL has shown statistically significant results against S. aureus [Table 3] and in 15 μL and 25 μL against E. coli [Table 4]. R. graveolens 30C in 10 μL, 20 μL and 30 μL has shown statistically significant results against S. aureus [Table 3] and in 15 μL, 20 μL and 25 μL against E. coli [Table 4]. R. graveolens 200C in 10 μL, 15 μL, 25 μL and 30 μL has shown statistically significant results against S. aureus [Table 3] and in 5 μL, 10 μL, 15 μL, 20 μL and 25 μL against E. coli [Table 4].

However, alcohol has shown more significant results in comparison to some homoeopathic medicines in certain volumes against both microbes [Tables 3 and 4].

DISCUSSION

The ultra-diluted nine homoeopathic preparations at different volumes, especially lower volumes, showed significant results against S. aureus (10 μL) and E. coli (15 μL) when compared to negative controls. This implies that homoeopathic medicines are useful for treating disorders caused by AMR bacteria such as S. aureus and E. coli at lower volumes.

Further, the macrodilution assay and disc diffusion assay will provide the minimum bactericidal concentration of homoeopathic medicines. Molecular docking is useful for the identification of compounds in homoeopathic medicines which bind to microbial structure for its inhibitory action. In vivo studies like acute toxicity studies in animal models and clinical trials may be carried out to confirm their effectiveness and mechanism of action of homoeopathic preparations.

CONCLUSION

The study suggests that ultra-diluted homoeopathic preparations at different volumes, particularly lower volumes, exhibited significant antimicrobial activity against S. aureus and E. coli compared to the negative controls. Therefore, it can be concluded that homoeopathic medicines may be useful in treating disorders caused by antimicrobial resistant bacteria such as S. aureus and E. coli at lower volumes.

Declaration of patient consent

Patient’s consent not required as there are no patients in this study.

Conflicts of interest

There are no conflicts of interest.

Financial support and sponsorship

Nil.

References

  1. . National Infection and Death Estimates for AR. . Centers for Disease Control and Prevention. Available from: https://www.cdc.gov/drugresistance/national-estimates.html [Last accessed on 2023 Jan 12]
    [Google Scholar]
  2. . A Textbook of Microbiology (2nd ed). Kolkata, India: New Central Book Agency Pvt Ltd; . p. :238-46.
    [Google Scholar]
  3. . Antimicrobial Resistance Research and Surveillance Network. Available from: https://www.main.icmr.nic.in/sites/default/files/guidelines/AMRSN_annual_report_2020.pdf [Last accessed on 2023 Jan 12]
    [Google Scholar]
  4. , , , , , , et al. Antiproliferative properties of polyketides isolated from Virola sebifera leaves. Phytother Res. 2008;22:127-30.
    [CrossRef] [PubMed] [Google Scholar]
  5. , , . In-vitro anti-microbial activity of methanolic extract of Atropa belladonna using the agar well diffusion method. World J Pharm Sci. 2014;2:654-6.
    [Google Scholar]
  6. , , , , , , et al. Evaluation of antioxidant and antimicrobial potential of two endangered plant species Atropa belladonna and Matricaria chamomilla. Afr J Trad Complement Altern Med. 2014;11:111-7.
    [CrossRef] [PubMed] [Google Scholar]
  7. , , , , . Antibacterial effects of Eucalyptus globulus leaf extract on pathogenic bacteria isolated from specimens of patients with respiratory tract disorders. Clin Microbiol Infect. 2006;12:194-6.
    [CrossRef] [PubMed] [Google Scholar]
  8. , . Chemical composition and antibacterial activity of Ruta graveolens L. (Rutaceae) volatile oils, from São Luís, Maranhão, Brazil. S Afr J Bot. 2015;99:103-6.
    [CrossRef] [Google Scholar]
  9. . Pocket Manual of Homoeopathic Materia Medica and Repertory: Comprising of the Characteristic and Guiding Symptoms of All Remedies (Clinical and Pathogenetic) Including Indian Drugs New Delhi, India: B. Jain Publishers Pvt Ltd.; . p. :270-71.
    [Google Scholar]
  10. . The Guiding Symptoms of our Materia Medica. Vol 2. New Delhi, India: B. Jain Publishers Pvt Ltd.; . p. :366-422.
    [Google Scholar]
  11. . A Dictionary of Practical Materia Medica. Vol 1. New Delhi, India: B. Jain Publishers Pvt Ltd.; . p. :720-1.
    [Google Scholar]
  12. . Homoeopathic Drug Pictures (2nd ed). New Delhi: B Jain Publishers Pvt Ltd; . p. :722.
    [Google Scholar]
  13. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically. In: Approved Standard-Tenth Edition. . Available from: https://www.clsi.org/media/1632/m07a10_sample.pdf [Last accessed on 2023 Jan 12]
    [Google Scholar]
  14. , , , , , , et al. Performance Standards for Antimicrobial Susceptibility Testing. Available from: https://www.clsi.org/media/2663/m100ed29_sample.pdf [Last accessed on 2023 Jan 12]
    [Google Scholar]
  15. , , , , , , et al. Methods For Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically In: Approved Standard-Eighth Edition. Available from: https://www.simpleshowoflove.weebly.com/uploads/1/4/0/7/14073276/agar_dilution_assay.pdf [Last accessed on 2023 Jan 12]
    [Google Scholar]
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