S al & urg ic ic d a e l M P f a o Utami, et al., J Med Surg Pathol 2016, 1:3 t l h Journal of Medical & Surgical o a n l o DOI: 10.4172/2472-4971.1000127 r g u y o J Pathology ISSN: 2472-4971 Research Article Open Access Study of Antimicrobial Activity of Black Cumin Seeds (Nigella sativa L.) Against Salmonella typhi In Vitro Amalia Tri Utami1*, Bogi Pratomo2 and Noorhamdani3 1Medical Program Study, Brawijaya University, Malang, Indonesia 2Internal Medicine Laboratory, Saiful Anwar General Hospital, Malang, Indonesia 3Microbiology, Brawijaya University, Malang, Indonesia *Corresponding author: Amalia Tri Utami, Medical Program Study, Brawijaya University, Malang, Indonesia, Tel: +628813477409; E-mail: [email protected] Received date: April 05, 2016; Accepted date: May 12, 2016; Published date: May 19, 2016 Copyright: © 2016 Utami AT, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Objective: To determine the effectiveness of extracts of black cumin seeds (Nigella sativa L.) as an antimicrobial against Salmonella typhi in vitro. And can determine minimum inhibitory concentration (MIC) and Minimal bactericidal concentration (MBC) from extracts of black cumin seeds (Nigella sativa L.) against Salmonella typhi. Design: This experimental study used post-test only control group design with four time repetition. Step one was cultivating bacteria in liquid medium with various concentration of extract, that was 40%, 42.5%, 45%, 47.5%, 50% with two control, extract control and bacterial control. Results: The MIC (Minimal Inhibition Concentration) is 45% concentration of extract. Step two was plating in NAP (Nutrient Agar Plate) medium. The MBC (Minimal Bactericidal Concentration) is 47.5% concentration of extract. The result of experiment was knew there are different average of Salmonella typhi colony from every group. The result experiment was analyzed by One Way Anova Test. The hypothesis test of MBC show significant differentiation, and then was continued with regression test. The conclusion of this study was the addition of extract concentration occurred reduction average of Salmonella typhi colony. Keywords: Antimicrobial; Extract ethanol; Nigella sativa L.; subsequent development of resistance of Salmonella typhi, several Salmonella typhi; In vitro countries reported a strain of multi-drug resistance (MDR) Salmonella typhi that is resistant to two or more antibiotics commonly used are Introduction ampicillin, chloramphenicol and cotrimoxazole. The development of MDR Salmonella typhi so fast in some countries resulting in mortality Bacterial of the genus Salmonella are capable of causing a large cases of typhoid fever in children increases, the experts seek another number of human infections, including typhoid fever, systemic alternative treatment for typhoid fever that fever down quickly, shorter infections, septicemia, and gastroenteritis varying clinically from treatment times and reduced relapse [4]. watery diarrhoea to dysentery [1]. The continued development of drug resistance of microorganisms to Typhoid fever is caused by microorganisms Salmonella enterica other microorganisms one that makes health organizations in the subspecies enterica serotype typhi (abbreviated as Salmonella typhi), is world were moved. Food Drugs Association (FDA) one of them, a systemic infectious disease, and is still a health problem in Indonesia. implementing an action plan against drug resistance with 4 stages, Endemic infectious diseases are growing in Asia, Africa, Latin namely, surveillance, development of a new drug product effective and America, the Caribbean and Oceania, including Indonesia, where natural, education, and research on resistance. The fourth stage is sanitation is poor. Although WHO declared typhoid fever not already underway FDA since 8 years ago [5]. including deadly diseases such as pneumonia, tuberculosis, malaria, measles and HIV/AIDS, but the epidemic of typhoid fever can be like Currently in Indonesia is developing a new paradigm in the field of in 1989, where 11 countries stricken with typhoid fever. In that year health, namely the use of natural herbs and traditional medicines. 10% of patients who died of typhoid fever. WHO said there were 20 Medicinal plants as natural resources have not been exploited and million deaths each year are caused by typhoid fever. WHO reported developed in depth is still very open to be researched and developed incidence of typhoid fever increased by 15.41% in 2008 [2]. Young under on finding an effective drug as an anti-microbial, especially in adult patients often experience severe complications such as bleeding typhoid fever. The use of herbal medicines is expected to be an and perforation of the bowel which often end in death [3]. Because of alternative for the community to prevent the side effects of antibiotic the high morbidity and mortality rates of typhoid fever, the various use [6]. Therefore, the need for alternatives to overcome these parties seek to resolve this problem. problems, logical, scientifically appropriate methods of "scientific knowledge". One possible approach is to use plants (herbs) that have In the last five years, clinicians in several countries observe any known or are still in the form of alleged anti-microbial effects that can cases of typhoid fever are severe and even fatal child, which was caused be used widely, with a good level of security or no side effects [7]. by Salmonella typhi strains resistant to chloramphenicol. In the J Med Surg Pathol, an open access journal Volume 1 • Issue 3 • 1000127 ISSN:2472-4971 Citation: Utami AT, Pratomo B, Noorhamdani (2016) Study of Antimicrobial Activity of Black Cumin Seeds (Nigella sativa L.) Against Salmonella typhi In Vitro. J Med Surg Pathol 1: 127. doi:10.4172/2472-4971.1000127 Page 2 of 4 10.4172/jmsp.1000127 Since one century, the Greek physician, Dioscorides, it has been bacterial growth while at a concentration of 47.5% is not contained doi: reported that black cumin or Nigella sativa or Black Seed to treat a bacterial growth. variety of ailments such as headaches, nasal congestion, dental pain, Based on this, researchers took concentration from 40% to 50% to and illness internist. During the golden age of Islam, by Al Biruni, the determine the value of MIC and MBC. The final concentration used in seeds of Nigella sativa used nutritional ingredients. In the treatment in this study is 40%, 42.5%, 45%, 47.5% and 50%. Researchers over the Greco-Arab or Greek-Tibb, from Hippocrates, Galen, and Avicenna, difference in the concentration range close up with the aim of MIC and Nigella sativa is a very valuable remedy in treating gastrointestinal MBC values obtained are more accurate. dysfunction and hepatitis described as a stimulant for the different conditions [8]. Religious factors also increase the confidence of his people to take advantage of Nigella sativa. In the hadith of Prophet The minimum inhibitory concentration (MIC) growth of Muhammad SAW said that black cumin can cure all diseases except Salmonella typhi death. Various studies show that in Nigella sativa contains essential oils Solution between black cumin seed extract and bacterial extracts (volatile oil). Essential oils are generally anti-microbial, anti- were made with a concentration of 40%, 42.5%, 45%, 47.5%, and 50% inflammatory [9]. Antimicrobials of black cumin seed extract in the for the then incubated at 37°C for 18-24 hours, then observed the form of active compounds such as: alkaloids, saponins, tannins, change of turbidity turbidity which is later will be able to know the melantin, derivatives terpenoids, and timokinon [7]. So far, studies level of minimum inhibitory concentration (MIC) black cumin seed have been conducted on the effects of Nigella sativa strains of the extract on the growth of Salmonella typhi. bacterium Vibrio colera and E. coli. The research that published in the journal Pharmaceutical Pakistan showed that more potent volatile oils to kill both bacteria compared antibiotics such as ampicillin and tetracycline [9]. Methodology Research conducted experimental research laboratory is using the tube dilution method to determine the concentration of extract of black cumin seeds (Nigella sativa L.), which can affect the growth of the bacteria Salmonella typhi isolates in vitro. The tube dilution method is to determine the MIC and MBC. This method includes two phases, phase of testing materials/extracts against isolates of Salmonella typhi in the media brain heart infusion broth to determine the MIC and the stage of planting in the medium Nutrient To Plate to determine MBC. There are five groups of treatment, coupled with germ control (0% extract material) and material control extract (100% Figure 1: Dilution results for Test Tube MIC against Salmonella extract material), with four repetitions. typhi. Result Shown in the Figure 1 that at a dose of 40%, 42.5%, is still turbid media, while at a dose of 45% compared with the media already clear Extract ethanol of Nigella sativa L. KK. KHM then obtained at a dose 45%. The end result of black cumin seed extract liquid is obtained black. From Figure 1 it can be seen that at the extract concentration of 0% Then extract that has obtained a suspension made by bacteria test BC (Bacterial Control) with four repetitions showed turbidity levels are (Salmonella typhi) that are