INTRODUCTION:

The shelf life of a product can be extended either by adding artificial preservatives or by taking hygienic measures during the manufacturing process. As the consumer trend today is towards preservative-free foods with a long shelf-life, industry is being forced to rethink its manufacturing methods. One of the main factors contributing to the increased appearance of multidrug resistant pathogenic strains that do not react to antibiotics in today’s society is the indiscriminate use of antibiotics. The standard course of action. Therefore, the necessity for discovering novel antimicrobials is still present. Alternative medications, such as natural or herbal medicines, are becoming more popular right now in addition to antibiotics and chemically synthesised treatments since they may have fewer side effects or toxicity due to their natural sources.¹

Curry leaves (Murraya koenigii) are a well-known leaf spice that are used in very small amounts for their distinctive aroma because they contain volatile oil and can enhance flavour. Digestion. In Asian cuisines, these leaves are frequently used to flavour meals. The leaves’ flavour and other characteristics are retained even after drying; they have a little aromatic, bitter, and acidic taste. In several ancient cultures, including Indian Ayurvedic and Unani prescriptions, curry leaf is also employed.²The Murraya koenigii plant produces leaves that are rich in minerals, vitamins, carotene, nicotinic acid, vitamins C and A, calcium, and oxalic acid. As well as Includes carbazole alkaloids, crystalline glycosides, iso-mahanimbin, koenigin, girinimbin, koenine, koenidine, and koenimbine. The leaves also contain the triterpenoid alkaloids cyclomahanimbine and tetrahydromahanimbine. Murraya koenigii leaves have been used to isolate several compounds, including murrayaline, pyrayafoline carbazole alkaloids, and murrayastine³ The Rutaceae family includes the significant leafy vegetable known as curry leaf (Murraya koenigii), which is indigenous to India and Southeast Asia. CurryLeaves are natural flavouring ingredients that have many beneficial health effects. They have a variety of therapeutic qualities, including anti-diabetic, antioxidant, anti-fungal, anti-microbial, anti-inflammatory, and Hepatoprotective and carcinogenic qualities. Among the plant’s many prominent pharmacological properties are its heart-healthy properties, anti-diabetic and cholesterol-lowering properties, antibacterial, antiulcer, antioxidative, cytotoxic, antidiarrheal, and phagocytic properties. Volatile oil makes up the majority of the fresh Murraya koenigii leaves’ chemical makeup. From Murraya koenigii’s stem bark and roots, carbazole alkaloids and triterpene have been extracted^3,4

Escherichia coli:

Gram-negative bacillus Escherichia coli (E. coli) is recognised to be a component of healthy intestinal flora but can also cause intestinal and extraintestinal disease in people. Numerous different E. coli strains have been identified, causing illnesses ranging from mild, self-limiting gastroenteritis to renal failure and septic shock. E. coli’s virulence makes it easier for it to overcome host defences and acquire antibiotic resistance. In this review, E. coli infections that result in extraintestinal illness will be separated from those that result in intestinal illness. The causative E. coli subtypes will be used to describe intestinal diseases, such as enterotoxigenic Escherichia coli (ETEC), enterohemorrhagic Escherichia coli (EHEC), also referred to as Shiga toxin-producing Escherichia coli (STEC), enteroinvasive Escherichia coli (EIEC), enteropathogenic Escherichia coli (EPEC), and enter (EAEC).⁵Clinical disease will be used to define extraintestinal

Fig. 1 E. coli

Etiology:

In addition to being found on the floors of hospitals and long-term care facilities, E. coli is a component of the commensal intestinal flora. The most prevalent gram-negative bacterium in the human gastrointestinal tract, E. coli, is not virulent in this environment. E. coli, however, can also result in pneumonia, bacteremia, peritonitis, urinary tract infections (UTI), and other conditions when found outside of the gastrointestinal tract.^6,7,8Numerous nosocomial infections, such as pneumonia linked to ventilators and catheter-associated urinary tract infections, are caused by E. coli (VAP).⁹In addition to undercooked meats, E. coli can also be found in water, on vegetables, and soil. When consumed by humans, pathogenic strains can result in intestinal disease.

Fig.2 E. coli

Staphylococcus aureus:

A common component of the body’s microbiota, Staphylococcus aureus is a Gram-positive, spherically shaped bacterium that belongs to the Bacillota and is frequently found in the upper respiratory tract and on the skin. It frequently exhibits catalase and nitrate reduction activity, and because it is a facultative anaerobe, it may grow without oxygen.¹⁰S. aureus is often a commensal member of the human microbiota, but it has the potential to transform into an opportunistic pathogen. It is a common cause of food poisoning, sinusitis, and various skin and respiratory infections, including abscesses. By creating virulence factors including strong protein toxins and the production of a cell-surface protein that binds and inactivates antibodies, pathogenic strains frequently aid in the spread of diseases. The emergence of antibiotic-resistant strains like methicillin-resistant S. aureus (MRSA) is a global problem in clinical medicine. S. aureus is one of the leading pathogens for deaths linked to antimicrobial resistance. There is currently no approved vaccine for S. aureus despite extensive research and development.

Fig. 3 S. aureus

S. aureus, which can be found as part of the normal skin flora, in the nostrils,^11,13and as a normal inhabitant of the lower reproductive tract of women, is thought to be long-term carriers in 20% to 30% of the human population.^11,12,14,15From minor skin infections like pimples,¹⁶ impetigo, boils, cellulitis, folliculitis, carbuncles, scalded skin syndrome, and abscesses to serious illnesses like pneumonia, meningitis, osteomyelitis, endocarditis, toxic shock syndrome, bacteremia, and sepsis, S. aureus can cause a variety of illnesses. It is one of the top five sources of hospital-acquired infections and frequently results in wound infections after surgery. In hospitals across the United States, S. aureus is the most common staphylococcal strain to infect patients.¹⁷ S. aureus infections have been linked to up to 50,000 deaths annually in the US.¹⁸

Fig.4 Skin Infection

Skin infection:

The most prevalent type of S. aureus infection is skin infections. Small benign boils, folliculitis, impetigo, cellulitis, and more serious, invasive soft-tissue infections are just a few of the different ways that this might present.^16,12 In people with atopic dermatitis, or more commonly, eczema, S. aureus is very common. The armpits, hair, and scalp are among the fertile, active areas where it is most commonly found. If lacerated, large pimples that develop there can make the infection worse. Staphylococcal scalded skin syndrome, which manifests in infants in its severe form, can result from this.¹⁹

Infection control:

The topical sanitizer ethanol has been shown to be effective against MRSA. To extend the time that ethanol has a cleaning effect, quaternary ammonium might be employed. Routine and terminal cleaning are necessary for the prevention of nosocomial infections. Nonflammable alcohol vapour has a benefit in CO₂ NAV-CO₂ systems since it doesn’t damage metals or plastics used in medical settings and doesn’t cause bacterial resistance. Though recently some veterinarians have discovered the infection can be spread through pets,²⁰environmental contaminations is thought to play a relatively less significant role in the spread of S. aureus (including MRSA), which typically spreads through human-to-human contact.²¹Therefore, putting a focus on fundamental hand washing procedures is useful in stopping its spread. Staff members who wear disposable aprons and gloves lessen skin-to-skin contact, which lowers the chance of transmission.

Murraya koenigii:

Murraya koenigii is sometimes referred to as Meethi neem or Curry Leaf/Kari pataa. It is a member of the Rutaceae family²². The plant is indigenous to India, Sri Lanka as well as a few more nations²³ It is extensively available and grown in India, as well as in Assam, Bengal, the Himalayas, Uttarakhand, the Indian state of Thiru-Kochi, and other places like the western Ghats, Asian moist forests, Bhutan, Nepal, Thailand, Pakistan, etc^22,24. The plant has a pleasant scent. It has a tiny shrub or a plant with a height of up to 6 metre’s^25,26. It is a plant that grows quickly and has dark green leaves.

Fig.5 Curry tree

Curry leaves include a variety of chemical components that have a variety of pharmacological and biological activities, nutritional benefits, and fragrant qualities. Some chemical components include caryophyllene, element, phallendence, and others because they give plants the capacity to stop food from going bad. Other substances include m-Cymene, -Terpinene, -Myrcene, Camphene, cis—ocimene, cis-Piperitol, Linalool, -Terpinene, and Terpinyl acetate. Thujene, eucalyptus oil, and caryophyllene oxide Additionally, the plant contains abundant amounts of girinimbiol and girinimbine (carbazole alkaloids), koenine, koenigine, and koenidine, as well as calcium, phosphorus, iron, thiamine (vitamin B1), riboflavin (vitamin B2), niacin (nicotinic acid), sitosterol, carotene, 1,4-methanoazulen^24,27,28.Murraya koenigii’s roasted leaves, combined with buttermilk to make a paste, are reportedly used as an anti-emesis. And regular oral consumption for stomach pain. The presence of vitamins (vitamins A, B, and C), calcium sources, and iron in plants makes plant parts useful for treating calcium and vitamin deficiencies as well. Curry leaves are also helpful for maintaining the black colour of hair. The paste made from the leaves is applied to boils and furuncles (painful, pus-filled bumps on the skin) to reduce pain and inflammation and prevent chataracts. The juice of roots Useful for treating kidney/renal pain^22,29Many researchers investigate According to reports, plants can be used to treat and cure a variety of diseases because they contain important chemical components in all of their sections. Which have a variety of medicinal properties, such as raw plant leaves that can be used to treat nausea, vomiting, and stomach pain. The plant’s leaves and roots are used to treat and cure conditions such as piles, anthelminthic, leucoderma (white patches on the skin), blood disorders, and they also have anti-inflammatory, analgesic, antibacterial, anti-cancer, antimicrobial, anti-oxidative, and useful properties for lowering cholesterol levels (hypolipidemic activity), anti-hypertensive, anti-diabetic, and other properties^30,25.

Biological source

Fig.6 Curry Leaves

The name of the species honours botanist Johann König. The Murray genus honours Johann Andreas Murray, a Swedish physician and botanist who passed away in 1791. Murraya koenigii is the curry leaves’ biological name as a result.³⁴

*Category*	*Botanical description*
Kingdom	Plantae
Subkingdom	Tracheobionta (vascular plant)
Superdivision	Spermatophyta (seed plant)
Division	Magnoliophyta (flowering plant)
Class	Magnoliospida
Subclass	Rosidae
Order	Sapindales
Family	Rutaceae
Genus	Murraya
Species	Murraya Koenigii

Chemical constituents of Murraya koenigii:

The plant extract made from Murraya koenigii contains a variety of organic compounds with different chemical compositions, including alkaloids, flavonoids, carbohydrates, and sterol. These compounds can be found in petroleum ether, ethyl acetate, chloroform, ethanol, and water.^{35,37,38,39,40} The principal chemical components are described. Numerous assays were carried out to validate the presence of phyto-constituents in the plant extract, including:

I. The presence of alkaloids was confirmed by using Mayer’s reagent, which shows formation of white or cream colored precipitates in the extract of Murraya koenigii.

II. Phenolic compounds were confirmed by formation of white precipitate by the addition of few drops of 5% lead acetate solution to alcoholic extracts of Murraya koenigii.

III. The presence of flavonoids is detected by Yellow coloration of filter paper by dipping in ammoniated alcoholic upon the extract.

IV. Presence of Saponins is considered when the extract showed honey comb like frothing formation after giving a shake with sodium bicarbonate.

V. The presence of proteins and free amino acids is indicated by the conducting the following tests i.e., Millons, Biurets and Ninhydrins test.

VI. Presence of sterol and triterpenes are indicated by alcoholic extract which was shaken with chloroform and few drops of acetic anhydride along with few drops of concentrated sulphuric acid from the side of the tube form the blue to brick red coloration.³⁶

The essential oil composition of Murraya koenigii was studied and then presence of D-Sabinene, D-α-Terpinol, di-α-phellendrene, D-α-pinene, caryophyllene and⁴¹ the property of Murraya koenigii oil is explained

Table: Essential oil property

Sl. No	Property	Value
1	Specific Gravity (25°C)	0.9748
2	Refractive Index (25°C)	1.5021
3	Optical Rotation (25°C)	+ 4.8
4	Saponification Value	5.2
5	Saponification Value after Acetylation	54.6
6	Moisture	66.3%
7	Protein	6.1%
8	Fat (Ether Extract)	1.0%
9	Carbohydrate	18.7%
10	Fibre	6.4%
11	Mineral Matter	4.2%
12	Calcium	810 Mg/100 G
13	Phosphorus	600 Mg/100 G
14	Iron Of Edible Portion	3.1 Mg/100 G
15	Carotene (As Vitamin A)	12 600 Iµ/100 G
16	Nicotinic Acid	2.3 Mg/100 G
17	Vitamin C	4 Mg/100 G
18	Thiamine And Riboflavin	Absent

Leaves:

Fresh Murraya koenigii leaves include between 61.77 and 66.2% moisture, 2.1-12.5% protein, 14.6-18.97% total sugar, 9.7-13.06% total ash, 1.35 and 1.82% acid insoluble ash, 1.35 and 1.82% alcohol soluble extractive, and 27.33 and 33.45% water extractive value.⁴²It explains the nutritional value. Alkaloids, flavonoids, and sterols have been extracted using solvents such as ethyl acetate, ethanol, petroleum ether, water, and chloroform during the manufacture of the extract. Koenigine, koenine, koenidine, and (-) mahanine were extracted from leaf extracts using acetone.⁴⁴Mahanimbine, isomahanimbine, koenimbidine, and murrayacine were separated from the hexane.⁴³It was discovered isomahanimbicine in petroleum ether⁴⁴

Nutrients value as per 100gm	Fresh curry leaf	Dry curry leaf
Protein	6g	12g
Fat	1g	5.4g
Carbohydrates	18.7g	64.31g
Calcium	830mg	2040mg
Iron	0.93mg	12mg
B-carotene	0.0031mg	0.059mg

Nutrients value of Murraya koenigii

Seed and fruit

Fig.7 Seed or Fruit of Curry Leaf

The furocoumarin lactone, carbazole alkaloids, glycolipids, phospholipids, and terpinene found in Murraya koenigii seeds. Additionally, it comprises 4.4% of the body’s total lipids, of which 5.1% are glycolipids, 9.5% are phospholipids, and 85.4% are neutral lipids. Neutral lipids contain modest amounts of diacylglycerols, monoacylglycerols, and sterols in addition to 73.9% triacylglycerol, 10.2% free fatty acids, and other lipids. Terpinene can be found in Murraya koenigii seeds in the following forms: terpinene, terpinen-4-ol, linolol,⁴⁵ ocimene, limblee, limbolee, and simbolee.^46-47 The pulp in the fruit is composed of 64.9% moisture, 16.8% total soluble solids, 9.76% total sugar, 9.58% reducing sugar, 0.17% non-reducing sugar, 13.35% vitamin C, 2.162% total ash, 1.97% protein, 0.082% phosphorus, 0.811% potassium (32,2), 0.166% calcium, 0.216% magnesium, 0.007% iron, and 0.00057% tannin.^{48,49,50,51,52}

Fig. 8 Stem and Bark of Curry Leaf

Stem and bark:

Murraya koenigii mature stem and bark contain a variety of chemical compounds, including glycolipids, phospholipids, coumarin galactoside, carbazole carboxylic acid, and carbazole alkaloids.

Root:

Fig.9 Root of Curry Leaf

Murraya koenigii roots contain a wide variety of bioactive substances. The root was extracted using petroleum ether and benzene.

Carotene

Girinimbine

Linalool

Tocopherol

Uses as antimicrobial agent:

Escherichia coli is one of the microorganisms that the root extract of Murraya koenigii with hexane, methanol, and chloroform extract is tested against (E. coli), Salmonella typhi, Staphylococcus aureus, a strain of Candida albicans, Trichophyton rubrum, and Aspergillus niger. The extract of this plant’s root in hexane, methanol, and chloroform has been found to have anti-microbial activity. The study also revealed that the methanol extract of this plant’s root was more effective than other extract chemicals in inhibiting microorganisms like Staphylococcus aureus and Trichophyton rubrum.

The aqueous extract of the root of this plant has been demonstrated in another study to be ineffective against microorganisms³¹

Use as antibacterial agent:

Bacillus cereus, Escherichia coli (E. coli), Staphylococcus aureus, and other bacteria are all susceptible to the antibacterial properties of Murraya koenigii plant extract. Typhi salmonella (S.typhi). The plant extracts of ethanol, methanol, and acetone demonstrated a notable effect against these bacterium strains.

The plant’s acetone extract had the greatest impact on Bacillus cereus, and the methanol extract had a higher level of inhibitory efficacy than the ethanol extract. Additionally, acetone extract demonstrated a higher inhibitory effect than methanol extract while methanol extract demonstrated less efficacy in inhibiting E. coli³²

Useful for skin pigmentation:

The cream made from the murraya koenigii plant’s leaves has the sun protection factor (SPF) property, which is excellent for protecting skin against sunburn. Erythema fights skin pigmentation (skin redness). Additionally, the cosmetic industry uses this plant to create formulations like cream, lotion, and soap as well as to add other chemicals to improve/ increase the efficacy of the finished product³³

Useful for Wound healing:

Plant ethanol leaf extract has the ability to heal wounds. The male albino rat used in the researchers’ investigation was slaughtered, and they applied the Tinctures in wounds The study found that the wound-related inflammation decreased over several days. Consequently, the research demonstrated that a plant leaf extract has wound-healing³².

Methodology:

The curry leaves were shade-dried for three days at room temperature, then ground in an electric blender

Methanol extraction:

The methanol extract was created by mixing ten grammes of curry powder with one hundred millilitres of a 70% aqueous methanol solution (w/v) that was covered with a cloth. Filter paper was maintained in a dark place at room temperature for two to three days after being shaken on a rotary shaker for 24 hours.

The final volume of the curry leaf methanol extract for the experiment was created by collecting the filtered supernatant and evaporating the solvent.

Ethanol extraction:

10gm of curry leaf powder were dissolved in 100 ml of ethanol and distilled water (8:2 w/v) to create the ethanol extract. With filter paper kept for 24 hours at room temperature and in a dark place. For the experiment’s final volume of curry leaf ethanol extract, the filteredsupernatant was collected, and the solvent was evaporated by incubation at room temperature for 48 hours.

Determination of antibacterial activity:

The paper disc was used to assess the antibacterial activity of the ethanolic and methanolic extracts of the curry leaf sample.⁵³

Diffusion technique:

The Whatman No. 3 filter paper discs, each measuring 6 millimetres in diameter, were immersed in methanolic and ethanolic solutions for 15 microliters at the following concentrations: 100 mg/ml, 50 mg/ml, 25 mg/ml, 12.5 mg/ml, and 6.25 mg/ml, respectively.⁵³

Agar well diffusion method:

A suspension of microorganisms in sterile The 0.5 was used to make normal saline. McFarland Guidelines. The bacteria’s turbidity Comparison of the suspension to the 0.5 McFarland standard solutions, followed by bacterial lawn culture using sterile Mueller-Hinton agar plates to suspend cotton swab and left in incubator for 15 minutes. The created methanolic and ethanolic mixture is then Filter paper-based discs were positioned on top of these For 24 hours, cultures and the plates were incubated at 37°C. To get consistent findings, the test was run three times for each extract, measuring the inhibitory zone around each disc in millimetres (mm). The four common antibiotics Amikacin, Ciprofloxacin, Amoxicillin, and Gentamicin were employed to contrast the curry leaves’ microbial inhibitory activity.

On agar plates with the bacteria to be tested inoculated, antimicrobial-containing filter paper discs were put down and allowed to incubate. Fully susceptible bacteria then showed a wide zone of growth inhibition surrounding the disc, but intermediately sensitive and resistant isolates showed a narrower inhibition zone. Showed no clear zones or reached the disc edge. Six bacterial species isolates, including Staphylococcus aureus and Streptococcus sp., were employed in this study. Representing gram-positive bacteria, and Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus sp., and Escherichia coli, which represent gram-negative bacteria.

Fig.10 E. coli

Fig. 11 St. auresus

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Received on 19.05.2023 Modified on 16.06.2023

Res. J. Pharmacology and Pharmacodynamics.2023;15(3):133-140.

DOI: 10.52711/2321-5836.2023.00024