17Ethnopharmacological relevance: Emergence of drug-resistant and multidrug-resistant 18Mycobacterium tuberculosis (Mtb) strains is a major barrier to tuberculosis (TB) eradication, as it 19 leads to longer treatment regimens and in many cases treatment failure. Thus, there is an urgent 20 need to explore new TB drugs and combinations, in order to shorten TB treatment and improve 21 outcomes. Here, we evaluate the potential of two medicinal plants, Artemisia annua, a natural 22 source of artemisinin (AN), and Artemisia afra, as sources of novel antitubercular agents. 23Aim of the study: Our goal was to measure the activity of A. annua and A. afra extracts against 24Mtb as potential natural and inexpensive therapies for TB treatment, or as sources of compounds 25 that could be further developed into effective treatments. 26 Materials and Methods: The minimum inhibitory concentrations (MICs) of A. annua and A. afra 27 dichloromethane extracts were determined, and concentrations above the MICs were used to 28 evaluate their ability to kill Mtb and Mycobacterium abscessus in vitro. 29Results: Previous studies showed that A. annua and A. afra inhibit Mtb growth. Here, we show 30 for the first time that Artemisia extracts have a strong bactericidal activity against Mtb. The killing 31 effect of A. annua was much stronger than equivalent concentrations of pure AN, suggesting that 32 A. annua extracts kill Mtb through a combination of AN and additional compounds. A. afra, which 33 produces very little AN, displayed bactericidal activity against Mtb that was substantial but weaker 34 than that of A. annua. In addition, we measured the activity of Artemisia extracts against 35Mycobacterium abscessus. Interestingly, we observed that while A. annua is not bactericidal, it 36 inhibits growth of M. abscessus, highlighting the potential of this plant in combinatory therapies 37 to treat M. abscessus infections. 38 Conclusion:Our results indicate that Artemisia extracts have an enormous potential for treatment 39 of TB and M. abscessus infections, and that these plants contain bactericidal compounds in 40 addition to AN. Combination of extracts with existing antibiotics may not only improve treatment 41 outcomes but also reduce the emergence of resistance to other drugs. 42 43
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