Indigenous Medicinal Plants use Sekhukhune, Limpopo, South Africa as Biofilm Inhibitors for the Mycobacterium Smegmatis

Indigenous Medicinal Plants use Sekhukhune, Limpopo, South Africa as Biofilm Inhibitors for the Mycobacterium Smegmatis

Dr. Jacobus Kori Madisha

Phytomedicine, Limpopo Education, 84 Limpopo Street, Modimolle, South Africa P.O Box 1777, Marble Hall 0450, South Africa

DOI: https://doi.org/10.51584/IJRIAS.2025.100500047

Received: 24 April 2025; Accepted: 03 May 2025; Published: 07 June 2025

ABSTRACT

The use of ordinary products to stimulate healthiness is as ancient as human sophistication. In contemporary years, the awareness of ordinary harvests derived from plants as ample foundations of naturally active complexes has determined their utilisation near the examination for novel biochemical products that can lead to further pharmaceutical formulations.  Opportunistic pathogens, escalation their virulence by acquiring resistance to orthodox antimicrobials, initiating infections, particularly in immunosuppressed hosts. Ethnobotanical information on these plants was obtained. Crude acetone extracts of 11 selected medicinal plants obtained in Sekhukhune, South Africa were screened for their ability to inhibit   Mycobacterium smegmatis    and a clinical strain resistant to first-line drugs and one second-line Rifampicin drug microplate assay to determine the minimum inhibitory concentration (MIC). Eleven plants were gathered and their acetone extracts evaluated for anti-mycobacterial  activity by the micro-dilussion  assay. All  extracts were tested for both antibacterial activity and biofilm inhibitory activity. The greatest encouraging herbal abstracts in combating  biofilm, particular their extraordinary capability to decrease it to small concentrations were the necessary oils removed from    Schotia brachpetala(B) ,Euphorbia tirucali(L), Eucalyptus camadulensis(L), Aloe marlothii (L), Elephantorrize elephantine( R ), Peltophorum africanum(B)

Keywords: Biofilm, Antibiotic, resistance, antibacterial,  Mycobacterium, Tuberculosis

INTRODUCTION

Microbial biofilms can offer a tireless and resistant setting for hazardous microorganisms inside the figure which donates to the growth of continuing inflammatory infections [8,10].  Biofilms have been originate in several structures and matters, comprising the central ear and greater breathing strip, verbal crack, circulatory coordination, lung, gastrointestinal, colon, urogenital coordination, maxilla, and undemanding muscle abrasions [11].Biofilm development establishes an different existence in which bacteria accept a multicellular comportment that enables and/or lengthens subsistence in diverse ecological positions. [17]. Biofilms system on living  and non-living shells mutually in the setting and in the healthcare situation [1].  A biofilm procedures when definite microbes (for example, some types of microorganisms) stick to the external of roughly entity in a humid location and begin to duplicate. [2].  The bacteria procedure an affection to the superficial of the entity by discharging a greasy, glue-like stuff. [14]. Four prospective enticements after the development of biofilms by microorganisms through contagion are measured: (1) fortification from detrimental environments in the swarm (2) confiscation to a nutrient-rich capacity, (3) consumption of supportive welfares , (4) biofilms customarily develop as biofilms [16].A shallow that make available humidity and nutrients is the perfect surroundings for biofilm improvement. Biofilms be able to be worthy, unscrupulous, or impartial [6]. Biofilms that are fragment of an ordinary setting are neutral, although biofilms that produce on exposed wounds subsequent contagion are detrimental. The systems of biofilm establishment is elicited and synchronised by quorum detecting, intimidating ecological settings, nutrient accessibility, hydrodynamic settings, cell-to-cell message, signalling cataracts, and tributary messengers. Integrating an basic cleanser or laundry detergent increases the efficiency of biofilm elimination paralleled to cleaning with bleach alone. [15]  Decolourize castoff at concentrations appropriate for nutrients communication shells ensures have some effectiveness on thermophilic bacilli and comparable biofilms, although effectiveness may be spasmodic. [3].  When patients do existent with stubborn fever, unwellness, pain and supplementary indications subsequent surgery and do not profit to antibiotic management they may have a bacterial biofilm contagion. [5].  Wounds that develop sick with biofilm may take drainage, hindered or partial restorative and an disagreeable odour. Amid numerous biofilm-associated infections and syndromes, disreputable instances comprise cystic fibrosis, otitis media, periodontitis, transmissible endocarditis [4]. Since the human perception, biofilms can be classified hooked on beneficial, neutral, and harmful. Harmful biofilms influence food security, development  of plant and animal infections, and intimidate medical arenas, making it crucial to develop operative and vigorous strategies to control harmful biofilms [6,7]. Antibiotics, antiseptics, and automatic elimination means are commonly used to remove biofilms and fight off related infections. Regular biofilm disruptors such as phytochemicals, sages and nourishments identical to garlic and kale, showcase capable capabilities in dropping biofilm development and addressing associated health challenges. [8].  Research have exposed garlic to be operative in distracting biofilm. Oregano is exclusively supportive for receiving rid of undesirable pathogens after the GI strip. It’s vigorous possessions, carvacrol, has been revealed to impede antibiotic resilient microorganisms, germs, vermin and moulds. In the main, a examination has reviewed the acetic acid properties, which is rich in vinegar, on the establishment of biofilm and discovered that it decrease the biofilm shaped by Staphylococcus aureus.

MATERIALS AND METHODS

Plant Material

Eleven herbal were gathered from Sekhukhune in the dumping area. Herbal was acknowledged at H.G.W.J. Schweickerdt Herbarium (PRU) at the University of Pretoria deposited a herbarium voucher specimen.

Preparation of Extracts

The plants abstracts were set with acetone (ACE) by method [19]  (Eloff,1998b)  and as explained previously [22]   (Agidew,2022 ).

Microorganisms Used

The bacterial strains namely activities of the were tested against Gram-negative bacteria, as well as Mycobacterium smegmatis (ATCC 1441).Bacterial cultures were grown overnight in Mueller Hinton (MH) broth (Sigma Aldrich, SA) and adjusted to McFarland standard 1.

Chemicals

Chemicals used in the assay were obtained from Highveld Biological, Johannesburg, South Africa and Doxorubicin was acquired from Pfizer, South Africa All organic solvents were of analytical grade and obtained from Sigma-Aldrich St. Louis, MO, USA. Müller Hinton agar and broth were from Sigma-Aldrich, India.

Antimycobacterial Determination of MIC (micro dilution)

The antibacterial activity of the herb extracts was studied by means of the micro dilution bioassay  as describe [19] with modification as describe by [20].MIC of the herbal abstracts was Identify. The lower the MIC is the enhanced is the activities . Antimicrobial activity of herbal abstracts take remained categorised as worthy (MIC< 0.1 mg/mL), reasonable (0.1 ≤ MIC ≤0.625 mg/mL) and insufficient (MIC >0.625 mg/mL) [19].

Antibiofilm development formation

The technique of [13] remained deployed to examine the plant extracts potential to inhibit development of microbial cell mass and attachment. The biomass was quantified with the changed crystal violet discolouration technique of [12].

Inhibition of pre-formed biofilm

The capability of plant abstracts to inhibit extra formation and or obliteration of cell quantity was also examined. The biofilm biomass was quantified by means of the changed crystal violet (CV) discolouration test [12].

Crystal violet staining assay

The method of Sandasi et al, 2008[13] was used for this assay, a modification of [12] Djordjevic et al, (2002). In brief, sterile distilled water was used to wash microtitre plates, air dehydrated and also oven dehydrated for 45 minutes in an oven set at 60 ⁰C. A 100 μl of 1% crystal violet was used to stain the wells of the plate, incubated for 15 min and later, the plates were cleaned three intervals with disinfected purified water to get rid of unreactive stain. At this level, biofilm was detected as florid ring by the side of the wells.  The quantitative assessment of biofilm development was determined by adding 125 µl of ethanol, this is to remove the stain in the wells. A 100 µl aliquot of the ethanol was withdrawn to a new sterile plate and the absorbance was measured using a microplate reader (BioTek) at 590 nm. The average absorbance was determined for each sample, and their respective percentage inhibition of biofilm calculated using the formula below (Sandasi et al., 2008) [13]:

Percentage (%) inhibition =       ODNegative control − ODExperimental × 100

                                         __________________________________________

                                                                             ODNegative control

RESULTS AND DISCUSSION

Table 1

Therapeutic plants assist as a possible opportunity for treatment innovation to fight infections and supplementary associated diseases. Acetone extracts Schotia brachpetala(B) ,Euphorbia tirucali, Eucalyptus camadulensis , Aloe marlothii (L), Elephantorrize elephantine, Peltophorum africanum(B). of  had MIC value of 0.0035 mg/mL to 1.6 mg/mL against Mycobacterium smegmatis. Acetone extracts of Schotia brachpetala ,Euphorbia tirucali, Eucalyptus camadulensis, Aloe marlothii, Elephantorrize elephantine, Peltophorum africanum(B). Senna italica  and Cannabis Sativa had MIC value of 2 and 2,5 mg/mL. Acetone was the best extractant, it extracted antibacterial agents which was shown by the lowest MIC values in all selected plants. The results obtained serve as a scientific validation for the use of the plants in traditional medicine for treatment of TB and additional respirational infection as well as their effectiveness in TB treatment innovation. Madisha [20]  and Khunoana et al. [21] also reported the antibacterial activity of A. Marlothii  and S. brachypetala against M.TB and M. bovis.

CONCLUSION

The plant species Schotia brachpetala(B) ,Euphorbia tirucali(L), Eucalyptus camadulensis(L), Aloe marlothii (L), Elephantorrize elephantine( R ), Peltophorum africanum(B) displayed active antiseptic activity concerning M. smegmatis validating their promising as foundations of TB treatment clue. Henceforth, the capable recommendation amongst the supposed highest classes of complexes in the abstracts and favourable activities in the study may lead in the prospect isolation and antiseptic assessment of the bioactive compounds. Further phytochemical and pharmacological investigations of these herbal are critical and noteworthy.

DECLARATIONS

Ethical approval

N/A

Consent for publication

I am the only author

Availability of data and material

 Available

Competing interest

No conflict of interest

Funding 

No funding

Authors’ contribution

One author work

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