The Application of Multiplate Resazurin Reduction Assay in The Screening for Anti-Mycobacterial Activity from Indonesian Medicinal Plants

Abstract

Tuberculosis (TB) is an airborne illness generated by Mycobacterium tuberculosis (Mtb), also one of the prominent infectious killers of adults worldwide. There is a pressing need to expand novel anti-mycobacterial drugs because of the increasing resistance of pathogenic mycobacteria to existing antibiotics. Native compounds acquired from microbial resources and medicinal cultivars have played an essential part as the origin of TB medications. The microplate resazurin reduction assay (MRRA) is generally utilized to assess natural and synthetic compounds for anti-mycobacterial activity. In our work, the MRRA method was employed to evaluate the antimycobacterial activity of extracts from curative plants using Mycobacterium smegmatis and Mycobacterium bovis BCG and to compare them to rifampicin as an anti-mycobacterial drug. The optimized MRRA utilized 2% aqueous DMSO and 62.5μg/mL resazurin as an indicator compound in 5% aqueous Tween 80. The optimal incubation time for M. smegmatis was 24h, and for M. bovis BCG was 48h. The methanolic plant extracts were acquired from various Indonesian medicinal plants known to have anti-mycobacterial activity. The various plant extracts exhibited anti-mycrobial activities confirmed by the MRRA assay. The MRRA method using M. smegmatis or M. bovis BCG as antimycobacterial targets offers a distinct advantage such as low-cost, rapid, and safe screening for anti-mycobacterial activity in a middle to high-through-putformat.

Therapeutic Potential of Punica granatum and Isolated Compounds: Evidence-Based Advances to Treat Bacterial Infections

“…Recent studies have indicated that extracts of the bark, leaves, stem, pulp, and whole fruit of pomegranate have efective antimicrobial activity, including against multiresistant strains [29,30,71]. Moreover, they demonstrated the antibacterial activity of pomegranate against bacteria commonly involved in upper gastrointestinal tract infections.…”

Composition and diversity of medicinal plant species and their utilization on agroforestry land at community forest bangkeng buki' in Bukit Harapan village Bulukumba regency

“…Each region has a distinctive plant utilization and is different from other regions. The utilization of different plants in each region is due to the diversity of plants in each region by Sari et al, [11]. One area that utilizes plants as medicinal materials by implementing an agroforestry system is the community living around the Bangkeng Buki' HKm located in Bukit Harapan Village, Gantarang District, Bulukumba Regency, South Sulawesi.…”

Screening of anti‐ Vibrio activity of marine fungal methanolic fractions to control vibriosis in white shrimp ( Litopenaeus vannamei )

“…The strong colour of most extracts could obscure the judgment of the turbidity level and OD measurement (Ardalani et al, 2021;Liu et al, 2007). Recently, a fluorescence-based assay using oxidation-reduction indicator resazurin has been successfully implemented for high-throughput screening (HTS) of antibacterial agents (Kim & Jang, 2018;Sari et al, 2019;Teh et al, 2017).…”

“…The strong colour of most extracts could obscure the judgment of the turbidity level and OD measurement (Ardalani et al, 2021; Liu et al, 2007). Recently, a fluorescence‐based assay using oxidation–reduction indicator resazurin has been successfully implemented for high‐throughput screening (HTS) of antibacterial agents (Kim & Jang, 2018; Sari et al, 2019; Teh et al, 2017). Resazurin (blue and not fluorescent) can be reduced by metabolically active cells to resorufin (pink and highly fluorescent) and upon further reduction to dihydroresorufin (colourless and not fluorescent).…”