Antibacterial Activity of Bandotan (Ageratum conyzoides L) Leaves Extracts Against Methicillin-Resistant Staphylococcus aureus

Antibiotic resistance is one of the biggest threats to global health. It is rising to dangerously high levels in all parts of the world (Aslam et al., 2018). Methicillin-resistance Staphylococcus aureus (MRSA) is a common cause of severe nosocomial infections (Choo & Hambers, 2016). It has developed resistance to numerous antibiotics caused by the misuse and overuse of antibiotics. The MRSA can hydrolyze almost any type of lactams, and its strains spread quickly, leading to a high mortality rate (Hu et al., 2019). World Health Organization (WHO)'s first global report on antibiotic resistance reveals that more than onequarter of S. aureus infections in the south-east Asia region are reported to be MRSA, which is home to a quarter of the world's population (Prestinaci et al., 2015). Therefore, alternatives antibacterial against MRSA infections is still sought-after investigation. Many antibacterial drugs were firstly isolated from natural sources (Rossiter et al., 2017). Many studies revealed that medicinal plants provide antibacterial compounds from its secondary metabolites (Gorlenko et al., 2020; Othman et al., 2019; Voravuthikunchai & Kitpipit, 2005). Indonesia is a rich archipelago with an abundance of natural plants to explore. One of them is bandotan (Ageratum conyzoides L.) plants, which grow in many Indonesian regions and are classified as tropical weeds (Kotta et al., 2020; Atisha & Mita, 2018). It is easy to find the weeds because it thrives in any garden and agricultural soils. It is also ubiquitous in disturbed sites and degraded areas (Marks & Nwachuku, 1986). The weeds are also noxiously regarded as harmful for crops, Antibacterial Activity of Bandotan (Ageratum conyzoides L) Leaves Extracts Against Methicillin-Resistant Staphylococcus aureus


INTRODUCTION
Antibiotic resistance is one of the biggest threats to global health. It is rising to dangerously high levels in all parts of the world (Aslam et al., 2018). Methicillin-resistance Staphylococcus aureus (MRSA) is a common cause of severe nosocomial infections (Choo & Hambers, 2016). It has developed resistance to numerous antibiotics caused by the misuse and overuse of antibiotics. The MRSA can hydrolyze almost any type of lactams, and its strains spread quickly, leading to a high mortality rate (Hu et al., 2019). World Health Organization (WHO)'s first global report on antibiotic resistance reveals that more than onequarter of S. aureus infections in the south-east Asia region are reported to be MRSA, which is home to a quarter of the world's population (Prestinaci et al., 2015). Therefore, alternatives antibacterial against MRSA infections is still sought-after investigation.
Many antibacterial drugs were firstly isolated from natural sources (Rossiter et al., 2017). Many studies revealed that medicinal plants provide antibacterial compounds from its secondary metabolites (Gorlenko et al., 2020;Othman et al., 2019;Voravuthikunchai & Kitpipit, 2005). Indonesia is a rich archipelago with an abundance of natural plants to explore. One of them is bandotan (Ageratum conyzoides L.) plants, which grow in many Indonesian regions and are classified as tropical weeds (Kotta et al., 2020;Atisha & Mita, 2018). It is easy to find the weeds because it thrives in any garden and agricultural soils. It is also ubiquitous in disturbed sites and degraded areas (Marks & Nwachuku, 1986). The weeds are also noxiously regarded as harmful for crops,

Antibacterial Activity of Bandotan (Ageratum conyzoides L) Leaves Extracts Against Methicillin-Resistant Staphylococcus aureus
Abstract Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of nosocomial infections throughout the world and can be lifethreatening as well. This study aimed to determine the antibacterial activity of Bandotan (Ageratum conyzoides L) leaves ethanolic extract against MRSA's growth. Ageratum conyzoides leaves were extracted by ethanol and screened for their phytochemical constituent. Ethanolic extracts of A. conyzoides leaves were evaluated for their potential antibacterial activity using disc diffusion assay. The minimum inhibitory concentration (MIC) value was determined using the agar dilution method. Phytochemical screening shows that the extracts contain alkaloids, flavonoids, saponins, tannins, and steroids or triterpenoids. Ageratum conyzoides leaves extract shows a 25.1 mm inhibitory zone at 12.5% extract concentration with MIC value equivalents to 4.46 x 10 -6 g of gentamicin. This study concludes that A. conyzoides leaves ethanolic extracts have potential antibacterial activity against MRSA.
but on the contrary, A. conyzoides have been known since ancient times for their therapeutic benefits (Garg et al., 2015

Plant material
Ageratum conyzoides plants were collected from the rice field at Limbangan, Garut, West Java, Indonesia. The plants were authenticated and determined at the Herbarium Unit, Department of Biology, Universitas Padjadjaran, Indonesia.

Extract preparation
Ageratum conyzoides leaves were washed and dried for   (Achmad et al., 2020;Chew et al., 2018;Amadi et al., 2012). Plants synthesized the secondary metabolites to protect them from predators such as herbivores, insects, and microorganisms. It could kill or inhibit microorganism growth via different mechanisms (Chew et al., 2018). The results of the antibacterial activity were revealed using the disc diffusion method. This is a standard qualitative assay to evaluate the antimicrobial activity of extracts or phytochemicals. Ageratum conyzoides leaves ethanolic extracts have demonstrated antibacterial activity against MRSA isolate. From Figure 1, it can be seen that the higher extract concentration produced a more expansive inhibition zone. The inhibition zone at 5% concentration of A. conyzoides leaves extract had the lowest average inhibition zone of 15.47 mm, while 12.5% had the largest average inhibition zone of 25.1 mm.

RESULTS AND DISCUSSION
Ageratum conyzoides leaves extract with 12.5% concentration, giving almost similar results compared to gentamicin, which was used as a comparative antibiotic.
This activity may be attributed to the rich tannins and flavonoid contents of A. conyzoides leaves. Flavonoids and tannins have been reported to possess antimicrobial activity due to their ability to complex with the bacteria cell wall and inactivate enzymes, microbial adhesion, and cell envelops proteins (Cowan, 1999). These results confirmed the evidence in previous studies that reported that the extract of A. conyzoides has potential antibacterial activity against S. aureus (Garg et al., 2015;Sugara et al., 2016). The quantitative analysis using the agar dilution method showed that at a concentration of 1%, 2%, and 3% of A.
conyzoides leaves extract still observed MRSA growth; meanwhile, the absence of MRSA growth can be seen at a concentration of 4% and 5% as shown in concentration was analyzed to determine its antibiotic dosage equivalency using the agar diffusion method as shown in Figure 2. Gentamicin was used as a reference standard antibiotic. Gentamicin is an aminoglycoside that inhibits bacterial protein synthesis by binding to its ribosomes (Krause et al., 2016). The standard dose of gentamicin is 3-6 mg/kg/day divided every eight hours to treat MRSA prosthetic valve endocarditis. Gentamicin is regularly added with rifampin for the first two weeks of treatment (Galar et al., 2019).

ACKNOWLEDGMENT
We are grateful for the support of the Chancellor of Universitas Garut for providing the research facilities.