P RODUCTION OF G REEN D ETTOL AS B ACTERIAL I NHIBITOR AND D ISINFECTANT

The aim of this research is production of disinfection (Dettol) contains herbal formula and study the efficacy of it for inhibition of pathogenic bacteria as and comparing with traditional Dettol efficacy. In this research, a herbal disinfectant and antiseptic was prepared similarly to the industrial Dettol and in terms of composition, except for the active substance (chloroxylenol), which was replaced by a plant extract (thyme) and active compound (thymol) separately. total of 50 samples hand swabs were collected randomly from children and housewives and student at university of Baghdad. The following species were isolated from swabs and identified by microscopic morphology, morphological features on culture media, and Vital Index of Traditional Environmental Knowledge(VITEK2)System and that were Staphylococcus aureus, Escherichia.coli, Bacillus cereus ,pseudomonas aeruginosa, Enterobacter cloacae . The results of study showed that the highest antimicrobial activity was belonged to thymol. while a lowest antibacterial effect was belonged to thyme. Highest value of minimum inhibitor concentration (mic) was (0.09) of thymol against staphylococcus aureus while a lowest mic value was (6.25) of thyme against Bacillus cereus the result of minimum bactericidal concertation (mbc) showed that thymol and Dettol were the same value (1.56) against Enterobacter cloacae.


INTRODUCTION
The Infectious diseases are a major cause of morbidity and mortality throughout the world. Nosocomial infections are illnesses that started or developed in a hospital or health-care facility. Nosocomial infections are on the rise and have become a major concern in hospital care, resulting in prolonged hospitalization, significant morbidity and mortality, and high expenditures, and excessive costs (9). The outbreak of the COVID-19 virus has affected countless people all over the world. Inadequate cleanliness and hygienic practices can lead to an increase in the infection rate during this outbreak (16) .Infection control procedures include the widespread use disinfectants in hospitals and medical centers to control the growth and elimination of microorganisms. Cleaning and sterilizing clinical equipment is also done with detergents and disinfectants. The antimicrobial activities of disinfectants are influenced by the disinfectant's composition, level of organic charge, temperature, dilution rate, and antimicrobial activity testing methodologies. The susceptibility of different types of microorganisms to antiseptics and disinfectants vary from one bacteria to another (10).
Many pathogenic organisms had become resistant nowadays for many manufactured antibiotics(15). Traditionally, resistance refers to instances where the basis of increased tolerance is a genetic change, and where the biochemical basis is known (8).
With the rising occurrence of medication resistance among common diseases, as well as the accompanying risk of chemotherapeutic medicines, finding an alternative to existing drugs has become critical. In ancient times, herbs bear been chronic within dense areas, together with nutrition, medicine, flavoring, beverages, cosmetics, etc...(14). Herbs with established pharmacological qualities may be the finest source of these alternative medications(18).
In Ayurveda and homoeopathy, medicinal plants with a broad spectrum of biological activity are used as a home cure for a variety of human illnesses. (7) Thyme (Thymus vulgaris L.) is an aromatic plants of the Mediterranean flora commonly used as spices and for medicinal purposes (4). Thyme is possessing antimicrobial, antifungal, antioxidative and antiviral properties (25).
Thymol is one of the most important components in thyme essential oil (17). Is an aromatic essence which is known as disinfectant in traditional medicine (24) . Thymol is a strong antimicrobial agent and is used as an antiseptic ingredient in some mouthwash's toothpastes on environment. There is a need for disinfection methods, which are rapidly effective, cost-effective, nontoxic, and that can be easily implemented (3). Most chemical disinfectants contain alcohol and usually brings adverse effects, which include allergic reactions and pathogens resistance (6) Global world is focusing now on use of herbal products because their considered are eco-friendly and more suitable to household applications than chemicals . (20) The aim of this study is to compare the efficacy of herbal disinfection using thymol and thyme in a green Dettol with commercially disinfectant Dettol.

MATERIAIS AND METHODS Collections of samples :
Total 50 skin swabs samples from the hands were collected randomly from children, housewives and student at university of Baghdad. the sample collected in Sterile moistened cotton swab sticks containing 5 ml of normal saline during the period from 1/9/2020 to 1/12/2020. To avoid drying, the cotton swabs were immediately moved to the laboratory (10).

Isolation and identification
Mac Conkey, mannitol, and blood agar were used to culture all of the samples. Subculture was done from each plate to separate and isolation colonies .The plates were incubated for 24h at 37 ℃ (13). These media used to distinguish and select the pure colonies according to (12).Several biochemical identification methods such as shape, Gram stain, Indole Test, Oxidase test, Catalase test, Citrate Utilization Test, Urease Test, Coagulase Test were conducted to identify the isolated bacteria (1, 11). Also, the diagnosis of the clinical isolates was confirmed by VITEK 2 compact. These identified cultures were transferred to nutrient agar slants for preservation and then stored in the refrigerator at 4°C (28) Preparation of herbal disinfection: The preparations of thymol and thyme disinfectant were conducted according to the Dettol procedure 1-ethanol 80 %( 30 ml) was mixed with 30ml castor oil, and stir well until mixed 2-Dissolved the 3.95gm of NaOH in a of 9 ml water, after dissolving it add on the mixture of (ethanol and oil). 3-Stirred the ingredients well until the mixture becomes homogeneous, leaved it for two hours, where the oil can be dissolved in water without turbidity. 3-In a clean beaker mixed 5ml of the pine oil with 5 gm of the thymol and stirred well. 4-The second mixture added to the first (alcohol and castor oil) stirred well. The volume of mixture reached up to 100 ml by adding distilled water 5-Added the 0.21g of caramel and move it well, ph it must be at 8

Preparation of microorganism suspension
The microorganism species maintained in BHI (brain heart broth) -20ºC.  (Fig. 1).  While a lowest antibacterial effect was with thyme against Bacillus cereus (6.25) mg/ml. as shown in (Fig.3 ).   (Fig.4).   values of F and p define the significance of the parameters, and the higher the value of F and the lower the value of p, the more significant [22]. In all the ANOVA statistical tests, the level of significance was p ≤ 0.05 or p ≤ 0.01 that suggesting that the test is considered to be statistically significant. According to the table (3)and(4) P-value was 0.000 for the significant differences among ability of (Dettol Thymol

Conclusions
From result obtained in this is study, it can be concluded that thymol have the broad activity against pathogenic microorganism and When used within commercial disinfectant (Dettol) ingredients as an antimicrobial agent instead of chloroxylenol and comparison with the results concluded that they were much for efficient than the chemicals compound (chloroxylenol) in the same tested concentration .The composition of thymol is natural and is less toxic than chloroxylenol, could be important motivators towards the pursuit of developing such products, the production of disinfection from natural compounds as an alternative to harmful chemical compounds, which could, eventually, safeguard and promote the public health.