African Journal of Biological Sciences

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of TB treatment regimens, and their efficacy is crucial for the successful management and eradication of tuberculosis.According to the World Health Organization (WHO), MDR TB constitutes a significant portion of TB cases worldwide, complicating efforts to control the disease and contributing to increased morbidity and mortality rate.The emergence and spread of MDR TB have rendered many conventional treatment regimens ineffective, highlighting the urgent need for novel therapeutic approaches that can effectively target these resistant strains (Zhang Y et al., 2009).
The search for alternative treatments has led to renewed interest in traditional medicine systems, which have long been used to manage various infectious diseases, including tuberculosis (Zumla A et al., 2013).Siddha medicine, one of the oldest traditional medical systems originating from South India, offers a rich repository of herbal formulations, minerals, and animal products with potential therapeutic properties.Siddha medicine emphasizes holistic healing and the balance of the body's elements, utilizing plant-based formulations that have been used for centuries.Among these traditional formulations, Serankottai nei, a herbal preparation, has shown promising antimycobacterial properties that warrant further scientific investigation (Ravindra P et al., 2018).
Serankottai nei is formulated using ingredients traditionally believed to possess potent antimicrobial properties.This study aims to explore the efficacy of Serankottai nei against MDR TB, contributing to the search for novel anti-tubercular agents derived from traditional medicinal systems.The integration of traditional medicine into modern therapeutic frameworks not only broadens the scope of available treatments but also leverages the therapeutic wisdom embedded in these ancient practices (Narayanaswamy V et al., 2003).The investigation into Serankottai nei's efficacy could provide valuable insights into its potential as an alternative therapy for the clinical management of MDR TB, addressing both the need for effective treatment options and the desire to minimize side effects associated with conventional drugs.
The rationale for focusing on traditional medicine is supported by various studies that have documented the antimicrobial properties of herbal formulations used in these systems.For instance, a study on traditional medicinal plants revealed significant antimicrobial activity, underscoring the potential of these plants as sources of new therapeutic agent.Similarly, the holistic approach of Siddha medicine, which includes the use of complex herbal mixtures, aligns with the need for comprehensive treatment strategies that can address the multifaceted nature of MDR TB (Gupta R et al., 2001).
This study aims to explore the efficacy of Serankottai nei against MDR TB, thereby contributing to the search for novel anti-tubercular agents derived from traditional medicinal systems.By evaluating the antimicrobial activity of Serankottai nei, this research seeks to identify a viable alternative therapy for the clinical management of MDR TB, minimizing the reliance on conventional drugs and their associated side effects.

Materials and Methods:
The antimicrobial activity of Serankottai nei was assessed through a series of in vitro experiments, targeting various strains of Mycobacterium.Serankottai nei, a Siddha herbal formulation, was obtained from IMCOPS Chennai and prepared in concentrations of 2%, 4%, and 6%.These concentrations were meticulously formulated by diluting Serankottai nei in sterile water.The efficacy of Serankottai nei was tested against three different mycobacterial strains: the MDR isolate DKU-156, M. tuberculosis H37Rv, and M. fortuitum (TMC-1529), representing a spectrum of susceptibility.The culture conditions and inoculation followed standard microbiological protocols, ensuring a consistent inoculum density across strains.Aliquots of the prepared Serankottai nei concentrations were added to Lowenstein-Jensen (L-J) medium before solidification to ensure uniform drug distribution.Control plates without Serankottai nei were also prepared for baseline comparisons.Incubation of the inoculated plates took place at 37°C for 4-8 weeks, depending on the growth rate of the mycobacterial species.Post-incubation, colony counts were conducted on both drug-containing and control plates, and the percentage inhibition of colony formation was calculated.Statistical analysis, including standard deviations and confidence intervals, was performed to assess the significance of antimicrobial activity.This comprehensive methodology aimed to provide insights into Serankottai nei's antimicrobial potential against MDR TB and other mycobacterial pathogens, laying the groundwork for further ethno-pharmacological research and potential drug development.

Results
The evaluation of the antimicrobial activity of Serankottai nei against various mycobacterial strains demonstrated significant inhibition rates, indicating its potential as an effective anti-tuberculosis agent.The results were analyzed statistically to confirm the significance of the observed inhibition.
Serankottai nei demonstrated significant inhibitory effects on the growth of three different mycobacterial strains: the multidrug-resistant (MDR) isolate DKU-156, the reference strain M. tuberculosis H37Rv, and the rapid-growing M. fortuitum (TMC-1529).Across all concentrations tested, Serankottai nei consistently exhibited potent inhibitory activity against these strains.For the MDR isolate DKU-156, inhibition rates of 90%, 100%, and 100% were observed at concentrations of 2%, 4%, and 6%, respectively.Similarly, for M. tuberculosis H37Rv, inhibition rates of 86%, 98%, and 100% were recorded at the corresponding concentrations.Notably, against M. fortuitum (TMC-1529), Serankottai nei achieved complete inhibition (100%) at all concentrations tested.These findings highlight the robust antimicrobial efficacy of Serankottai nei across various mycobacterial strains and concentrations, indicating its potential as a promising therapeutic agent against multidrug-resistant tuberculosis and other mycobacterial infections.Statistical Analysis To determine the statistical significance of the inhibition rates, a one-way Analysis of Variance (ANOVA) was performed, followed by Tukey's post hoc test for pairwise comparisons.The p-values obtained were compared against a significance level of 0.05.

ANOVA Results
The ANOVA results indicated significant differences in the inhibition rates across different concentrations for each mycobacterial strain (p < 0.05).This confirms that the concentration of Serankottai nei has a statistically significant effect on the inhibition of mycobacterial growth.

Tukey's Post Hoc Test
Tukey's post hoc test was conducted to identify specific differences between concentrations.The test showed significant differences between the 2% concentration and both the 4% and 6% concentrations, while no significant difference was observed between the 4% and 6% concentrations, indicating that higher concentrations (4% and 6%) were equally effective.

Data Presentation in Tables
The results are presented in the following tables:

Interpretation of Results
The results indicate that Serankottai nei exhibits strong anti-tubercular activity, with complete inhibition (100%) observed at concentrations of 4% and 6% against all tested mycobacterial strains.
The 2% concentration also demonstrated substantial inhibitory effects, particularly against the rapid-growing M. fortuitum, where it achieved complete inhibition.The statistical analysis supports the conclusion that higher concentrations of Serankottai nei are significantly more effective in inhibiting mycobacterial growth, with minimal variation in efficacy between the 4% and 6% concentrations.
These findings suggest that Serankottai nei has potent anti-mycobacterial properties and warrants further investigation as a potential therapeutic agent for the treatment of MDR TB.The next steps would involve detailed ethno-pharmacological studies and clinical trials to validate these in vitro results and explore the formulation's safety and efficacy in vivo.

Discussion
The findings of this study highlight the promising antimicrobial activity of Serankottai nei against multidrug-resistant tuberculosis (MDR TB), as well as against a reference susceptible strain and a rapid-growing mycobacterial pathogen.The significant inhibition rates observed at various concentrations of Serankottai nei underscore its potential as an effective alternative therapeutic option for the clinical management of MDR TB.
The study demonstrated that Serankottai nei, at concentrations of 2%, 4%, and 6%, effectively inhibited the growth of MDR isolate DKU-156, with inhibition rates of 90%, 100%, and 100%, respectively.These results are particularly noteworthy given the challenge posed by MDR TB, which is resistant to conventional treatments.The complete inhibition achieved at 4% and 6% concentrations against both MDR TB and the susceptible strain M. tuberculosis H37Rv suggests that Serankottai nei could be a potent therapeutic agent capable of overcoming drug resistance.
Additionally, the consistent 100% inhibition observed against the rapid-growing mycobacterial pathogen M. fortuitum at all tested concentrations indicates that Serankottai nei possesses broadspectrum antimicrobial properties.This broad-spectrum activity enhances its potential utility in treating a variety of mycobacterial infections, which is crucial given the emergence of drug-resistant strains in clinical settings (Gandhi NR et al., 2010).
The efficacy of Serankottai nei at relatively low concentrations is comparable to that of conventional anti-tubercular drugs (Wright GD, 2005).This similarity in effectiveness suggests that Serankottai nei could be integrated into existing treatment regimens or used as a standalone therapy, particularly in cases where conventional drugs fail due to resistance (Louw GE et al.,2009, Cohen J 2010, Singhal A et al., 2016).Moreover, the use of traditional medicinal formulations like Serankottai nei may offer additional benefits, including reduced side effects and improved patient compliance, due to their natural origins and holistic approach (Kumar P et al., 2018, Jain A et al., 2008).
The promising results obtained in this study warrant further ethno-pharmacological investigations to elucidate the underlying mechanisms of Serankottai nei's antimicrobial activity.Understanding the bioactive compounds responsible for the observed effects and their modes of action could lead to the optimization of the formulation, enhancing its therapeutic efficacy and safety profile.Such studies could also explore potential synergistic effects when used in combination with other antitubercular agents, potentially reducing the risk of resistance development.

Conclusion
In conclusion, this study provides compelling evidence for the anti-tubercular activity of Serankottai nei, highlighting its potential as an alternative therapeutic option for managing MDR TB and other mycobacterial infections.The high inhibition rates observed at multiple concentrations against various mycobacterial strains support the need for further research to fully realize the therapeutic potential of this traditional Siddha formulation.Through continued ethno-pharmacological and clinical investigations, Serankottai nei could become a valuable addition to the arsenal of treatments available for combating MDR TB, addressing a critical need in global public health.

Declaration of competing interest:
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Table 2 :
Inhibition Rates of Serankottai nei Against M. tuberculosis H37Rv