natural inhibitors of c. difficile

Introduction

Since centuries ago, hundreds of plants worldwide are used in traditional medicine as natural alternative treatments for different diseases and bacterial infections. Some of these tested in vitro against different diseases and some of them showed good results but the efficacy of these herbs still unknown against other different diseases. Conventional drugs usually show effective antibacterial activity for bacterial infections but the increasing resistance against these antibiotic raised need for new solutions. Although natural products are not necessarily safer than known synthetic antibiotics, some people prefer to use herbal medicines.

Garlic

Garlic is one of the food plants, which have massive interest throughout human history as a medicinal herb, and has been used for thousands of years as medicinal properties; exploration into garlic mode of action are relatively recent. Garlic has a wide spectrum of actions; it is known as antibacterial (Tessema, Mulu et al.,2006; Cai, Wang et al.,2007; Eja, Asikong et al.,2007; Friedman, Henika et al.,2007; Tsao, Liu et al.,2007), antiviral (Liu, Fang et al.,2004; Zhen, Fang et al.,2006), antifungal (Ogita, Nagao et al.,2007) and antiprotozoal (Hafez and Hauck 2006), and also has beneficial effects on the cardiovascular (Gromnatskii, Sereditskaia et al.,2007), Moreover, garlic also has been reported to reduce blood lipids (Aksenov, Kaplun et al.,2007), and have anticancer effects (Herman-Antosiewicz, Powolny et al.,2007) and play a role on the immune systems (Dorhoi, Dobrean et al.,2006). Renascence in the use of natural herbal alternatives has brought the use of medicinal plants to the forehead of pharmacological investigations, and hundreds of new drugs are being discovered. Louis Pasteur was the first to describe the antibacterial effect of onion and garlic juices in this century (Harris, Cottrell et al.,2001). In 2001 garlic raw juice was found to be effective against many common intestinal bacteria pathogens, which are responsible for diarrhea in humans and animals (Sivam 2001). Garlic is effective even against some strains which known as highly resistant to their usual antibiotic (Mutsch-Eckner, Erdelmeier et al.,1993).

Analysis of steam filtrations of well crushed garlic pods carried out more than a century ago illustrated the presence of a selection of varied allyl sulfides. Nevertheless, it was only in 1944 that Cavallito and his group, were able to isolate and identify the component that was responsible for the antibacterial action of crushed garlic (Harris, Cottrell et al.,2001). The compound was recognized to be an oxygenated form of sulphur which was termed as allicin (from the Latin name for garlic plant, Allium sativum). Pure allicin is a changeable molecule that is poorly dissolvable in aqueous solutions with typical odor of freshly crushed garlic (Harris, Cottrell et al.,2001). In 1947 final proof of the chemical structure of allicin was revealed (figure 1), when it was shown that allicin could be synthesized by mild oxidation of diallyl disulfide (Harris, Cottrell et al.,2001). The argue on the presence or absence of allicin in crushed cloves was resolved after Stoll and Seebeck identified, isolated, and synthesized an oxygenated sulfur amino acid which is present in garlic cloves and named alliin (Harris, Cottrell et al.,2001) (figur1). Alliin was established as the stable precursor which gets transformed to allicin by the action of an enzyme known as alliinase which is also known to be present in cloves(Harris, Cottrell et al.,2001). Numerous investigators studied the amounts of alliin and allicin present in different strains of garlic (Harris, Cottrell et al.,2001).

In 1991 Hughes and Lawson showed that the antimicrobial activity of garlic is completely abolished when allicin is removed from the extract (Lawson, Wang et al.,1991; Lawson, Wood et al.,1991). the antibacterial activity of allicin is exhibited by immediate and total inhibition of RNA synthesis and partially by inhibiting the DNA and protein syntheses (Feldberg, Chang et al.,1988; Mutsch-Eckner, Erdelmeier et al.,1993). The antibacterial activity of allicin against bacteria depends on the structural differences of the bacterial strains which may play a role in the bacterial susceptibility to garlic constituents (Sivam 2001).

Tea tree oil

Melaleuca alternifolia or tea tree, indigenous to northern New South Wales and southern Queensland and Australia (de Groot and Weyland 1992). Tea tree oil extracted by steam distillation from grown trees, with 100 tea tree growers in Australia producing over 500 tons of tea tree oil per annum (2004/2005). Need for tea tree oil increased by 25% in 2003/2004 and 13% in 2004/2005 (Rutherford, Nixon et al.,2007). Tea tree oil is widely used becoming widespread in Europe and North America; however, only a small amount of the oil is produced outside of Australia (Veien, Rosner et al.,2004). It is currently known that tea tree oil is marketed as a 'natural' topical antimicrobial and anti-inflammatory oil prepared at different concentrations. As use has increased, there were reports about the implication of tea tree oil in causing skin reactions including Allergic Contact Dermatitis (Crawford, Sciacca et al.,2004).

Myrrh

The genus Commiphora comprises over 200 species, most of which are confined Myrrh. They are small trees with thorny branches. It is found in the dry and arid regions of a southern Arabia, Nubia (Egypt and Sudan), Djibouti, northeast Somalia, and the Arabian Peninsula of southwest Asia (Yemen) and India (Hanus, Rezanka et al.,2005) (Figure:2). It has characteristically bitter taste. C myrrha is also known by the name of Commiphora molmol. Other familiar names include "Arabian myrrh," "Karam," "Morr Higazi," "murr," "guggul" (India), "makkul," "malmal" (Somalia), "mo yao" (China), and "bola (Omer, Adam et al.,1999). In the 5th century BC, Herodotus wrote: "Arabia is the only country which produces frankincense, myrrh, cassia, and cinnamon (Miller and Goodell 1968).

It is an oleo gum resin acquired from the stem of the plant (Figur 2.1). It is a safe, natural substance which is approved by the US Food and Drug Administration. It is a homogeneous mixture of materials and Samuge cameras and unmanned oil.

Myrrh traded throughout the Middle East at least since 1500 BC. It is documented that Egyptian used the myrrh before 3000 years BC and called it as "the tears of Horus." The "Papyrus Ebers" one of Egypt's oldest formulary of prescriptions, dating 1500-1700 BC, cited the use of myrrh to mummify and mummify and to stop bleeding , cremation and funerals as will as in the treatment of wound and skin sores (Michie and Cooper 1991). In the 4th century BC, Hippocrates referenced myrrh in his writings as useful in both medicine and as"healing" incense. Another Greek physician, Dioscorides, prescribed myrrh for the treatment of cough and infections. Ancient Romans burned myrrh to cover the odor emitted from the cremation of cadavers. Myrrh, already familiar in China and was formally acknowledged in herbal medicine books for treating various skin diseases. One such medical reference is the Kaibao Bencao or Materia Medica of the Kaibao Era, 973 AD (Miller and Goodell 1968). Myrrh had a wide application described in early Sumerian inscriptions describing treatment for infected teeth and warms (Michie and Cooper 1991). The Hippo crating writings (4th century BC) contain about 54 references to myrrh and descriptions of the use of its incenses in the treatment of wound sepsis. In Greek Myrrh or its oil was prescribed for teeth, moth, cough and eyes infection in children (Miller and Goodell 1968).

Myrrh's antibiotic affect had been successfully utilized before the first century BC to brevet the fermentation of wine to vinegar a central challenge of the technology of the day 'Aromatic' wines. Both Roman and Greek used Myrrh for treat from snake bite and that what they still use it for in East Africa (Michael J. O'DOwd 2001). Myrrh has been used in Middle Eastern medicine for treatment of infected wounds for thousands of years. It is also familiar to many cultures as incense used in religious rituals. Myrrh is a Jewish holy oil, and is mentioned in the first books of the Muslim, Jewish, and Christian holy texts. This herb is best known through the story of the Three Wise Men (Magi) delivering myrrh, gold, and frankincense for the baby Jesus. Today, crude myrrh is dispensed throughout eastern Africa and Saudi Arabia as an anti-inflammatory and anti-rheumatism drug (Iwu, 1990).

In experimental studies on Swiss albino mice, myrrh showed a potent cytotoxic activity against Ehrlich solid tumour cells. Myrrh also showed antischistosomal and other antiparasitic activity in animal and human and have found it to be safe and effective (Massoud, Metwally et al.,2004; Soliman, El-Arman et al.,2004; Fathy, Salama et al.,2005; Hamed and Hetta 2005; Al-Mathal and Fouad 2006; Fenwick and Webster 2006). In Egypt study showed that Mirazid (purified extract of myrrh) is effective as fasciolicidal drug (Massoud, El-Kholy et al.,2004)

It was also revealed that myrrh has good effect in healing deep wounds infections (Lotfy, Badra et al.,2006). Myrrh has antifungal and antibacterial activity against standard pathogenic strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. It was shown that myrrh also has local anaesthetic activity (Lotfy, Badra et al.,2006).

In alleviating inflammatory diseases such as arthritis and rheumatism, chemicals that derived from plants including grape, turmeric, and some essential oils such as clove, rosemary, mint, lavender, myrrh, and pine have been patented and used as mixed preparations. These plants are the most potent in curing inflammatory diseases (Darshan and Doreswamy 2004; Tipton, Hamman et al.,2006).

Myrrh tincture is used to treat amenorrhea and dysmenorrhea and to relieve some of the uncomfortable symptoms of menopause. The British Herbal Pharmacopoeia reported antiseptic action. It has also been shown to have deodorizing, disinfecting, and granulation-promoting properties.

Ginger

Ginger, the rhizome of Zingiber officinale, is one amongst the most extensively used species of the ginger family (Zingiberaceae) and is used as a common flavoring agent in a variety of drinks and foods (White 2007). Since 2,500 years, ginger has a long history in medical use in China and India for conditions such as nausea, headaches, colds, and rheumatism. Ginger was mentioned in Chinese medicine and is claimed to treat cold extremities, warm the body, improve a slow pulse, and strengthen the body after blood loss (Park and Pezzuto 2002). Steam filtration of powdered ginger produces ginger oil, which contains a high proportion of sesquiterpene hydrocarbons, predominantly zingiberene (Ranganna, Govindarajan et al.,1983). The compound 6-gingerol appears to be responsible for its characteristic taste (Govindarajan 1977). The National Center for Complementary and Alternative Medicine (NCCAM) has estimated the facts about ginger available in the studies, classifying the reports from "suggestive" (for short-term use of Ginger for safe relief from pregnancy related nausea and vomiting), to "mixed" (when used for nausea caused by motion sickness, chemotherapy, or surgery), and to "unclear" for treating rheumatoid arthritis, osteoarthritis, or joint and muscle pain) (Hoffman 2007).

Black seed oil

The black seed or Nigella Sativa, a plant from the Ranunculaceae (buttercup) family is known for over two thousand years and has been traditionally used by different cultures throughout the world as a natural medicine for many diseases and to improve health in general (Black Seed: Nature's Miracle Remedy, Amazing Herbs Press, New York, NY (2003)). The black seed was used by the old Egyptians who described it as a panacea (cure for problems and diseases) (P. Schleicher and M. Saleh, Black seed cumin: the magical Egyptian herb for allergies, asthma, and immune disorders, Healing Arts Press, Rochester, Vermont (1998), p. 90.). The black seed was also known by the Romans who called it Greek Coriander and used it as a dietary supplement. In the first century, the Greek physician Dioscoredes wrote that the black seed was used to treat different symptoms such as nasal congestion, toothache, headaches, and intestinal worms.

The black seed is also mentioned in the Bible in Isiah 28:25-27 as the 'fitches'. Ibn Senna, who wrote the great medical treatise 'The Canon of Medicine', stated that the black seed 'that stimulates the body's energy and helps recovery from fatigue' (Salem 2005). The black seed is planted in Turkey, Russia, Egypt, Oman, Arabia, Ethiopia, Far East, Middle East, Bangladesh, India, Germany, France, and the Mediterranean Basin. Nigella Sativa is known commonly in Arabic as Habbat-ul-Baraka (blessed seed) and in English as Love in the Mist. Since 1959, over 200 studies have been carried out at different international universities and articles were published in various scientific journals. These studies have shown excellent results supporting its traditional uses. The Nigella Sativa seed itself contains many esters of unsaturated fatty acids and its active ingredient is crystalline nigellone. It also contains 15 amino acids (Bhatia and Bajaj 1972; Ghosheh, Houdi et al.,1999; Chun, Shin et al.,2002), fatty acids including linolenic, oleic, volatile oils, and dietary fibre, as well as carbohydrates, minerals such as sodium, iron, calcium, and potassium(Figure :4) (Mahmoud, El Abhar et al.,2002; Ramadan and Morsel 2002).

There are researches on the black seed in which shown that black seeds can be used as an anti-tumor (Shapiro, Wu et al.,1993; Islam, Begum et al.,2004), anti-inflammatory, anti-histaminic (Chakravarty 1993; el Dakhakhny, Abdel el-Latif et al.,1996; Al Majed, Daba et al.,2001; Gilani, Aziz et al.,2001; Kalus, Pruss et al.,2003), anti- microbial (Agarwal, Kharya et al.,1979; Hanafy and Hatem 1991; Salem and Hossain 2000), anti-viral (Reynolds, Rahija et al.,1993; Moro, Lloyd et al.,1999), and anti-bacterial (Hanafy and Hatem 1991).

There was no information available on mustard oil, red chilly oil, sesame oil, black seeds oil, hemp oil, red onion, green onion, leak, green chilly, and clove.

Aims

Due to increasing of C. difficile's resistance to antibiotics, it is mandatory to find new antibacterial products. To find cheap antibacterial products to replace antibiotics. To go back to the herb medicine which was used in the past. To avoid the side effect of antibiotics.

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