# _Pipper nigrum_ Fruit Extract as an Antibiotic Resistance Reversal Agent in MDR Bacteria
###### Abstract  
Antibiotic resistance development and spread in clinical pathogens is an immense threat that has already outpaced the discovery and development of novel and more effective antibiotic drugs. Recently the focus has been shifted to medicinal plants as novel therapeutic options for reversing antibiotic resistance by targeting different resistance mechanisms. _Pipper nigrum_ is a plant that has the potential to reverse antibiotic resistance and increase the efficacy of the current drugs. In the present study, seven different antibiotics, clindamycin, gentamicin, levofloxacin, amikacin, tiegcycline, imipenem, and tetracycline, were used against antibiotic-resistant _Staphylococcus aureus_ and _Salmonella typhi_. Antibiotic resistance reversal analysis was tested by the AST disc method. Increased zones of inhibition of _S. aureus_ by four antibiotics, clindamycin (9 mm), gentamicin (7 mm), levofloxacin (9 mm), and amikacin (9 mm) were recorded after using _P. nigrum_ extract. In addition, the use of _P. nigrum_ extract also increased the zone of inhibition of _S. typhi_ with amikacin (11 mm), gentamicin (10 mm), tiegcycline (9 mm), levofloxacin (11 mm), and imipenem (10 mm). This study suggests that _P. nigrum_ extracts can be used as natural antibiotic resistance reversal agents that increase the effectiveness of current antibiotics and can reverse antibiotic resistance. Pipper nigrum; antibiotic resistance reversal; multidrug-resistant bacteria; antibiotics 20220202020220220202022020220220220220220220220  
The semi-synthetic method of drug development can generate several analogs in which functional groups of natural products are modified. This method provides the best results, with low toxicity and high bioactivity [12]. However, semisynthetic or unnatural agents cannot always be recommended for treating bacterial disease. Various plant extracts and essential oils are reported to be useful as natural antimicrobial agents [13; 14; 15]. Many plant-derived compounds are reported to exhibit promising results against MDR bacteria and can reverse antibiotic resistance. However, despite the antimicrobial activity, many plant extracts and essential oils (EO) cannot be considered clinically significant alone [9]. These antimicrobials, if administered with antibiotics, can decrease antibiotic resistance. The combination of drug and plant extract or essential oil can enhance clinical outcomes. Plant extracts can be used to reverse antibiotic resistance. Plasmid-mediated antibiotic resistance is minimized by eliminating plasmids from bacteria and this process is called as plasmid curing [13]. Secondary compounds, like quinones (consisting of bioactive compounds major class), that are derived from plants show activity against resistance and have the ability to eliminate plasmids [16]. Herbal extracts of _Piper nigrum_, _Zingiber officinal_, _Cinnamoun verum_, _Nigella sativa_, _Plumbago zyelnelica_, etc. contain phenol (eugenol), saponin, naphthoquinones, flavonoids, tannins, and alkaloids, which can be used as the plasmid curing agent [17]. _P. nigrum_, a member of the family Piperaceae, is used for chills, skin disorders, fever, muscle aches, flu, pain relief, asthma, diarrhea, and obesity [18]. _P. nigrum_, known as \"the king of spices\", helps to decrease inflammation, regulate lipid metabolism and decrease the risk of cardiovascular diseases [19]. North-Eastern people use it as herbal medicine for diseases such as headache, cold, fever, cough, diabetes, skin disease, leprosy, piles, pneumonia, and stomach-ache [20]. Piperine, a primary active compound of _Piper nigrum_, has been reported to increase drug bioavailability. The antibacterial activity of _P. nigrum_ petroleum ether extract was confirmed against gram-positive and gram-negative bacterial strains. On fractionation, active petroleum ether extract yielded five compounds, including the isopiperolein B [21], 2E, 4E, 8Z-N-isobutyleicosatrienamide [22], trachyone, pellitorine, and persumidiene. For this reason, these extracts of _P.