CACTUSNET

NEWSLETTER

FOOD AND AGRICULTURE ORGANIZATION OF THE UNITED NATIONS

INTERNATIONAL CENTER FOR AGRICULTURAL RESEARCH IN THE DRY AREAS

FAO-ICARDA INTERNATIONAL TECHNICAL COOPERATION NETWORK ON CACTUS

UNIVERSIDAD NACIONAL DE SANTIAGO DEL ESTERO ARGENTINA

CACTUSNET. Issue 11. December 2007 International Center for Agricultural Research in the Dry Areas/ North Africa Program Universidad Nacional de Santiago del Estero, Argentina

Table of content
Editorial Scientific and technical contributions
Ali Nefzaoui, Monica Nazareno, and M. El Mourid. Review Of Medicinal Uses Of Cactus. Mnica A. Nazareno and Evangelina Gonzlez. Antioxidant Properties Of Cactus Products. Helmut Sinzinger. Antihyperlipidemic Effect Of Cactus Products. Catherine Thimonier and Karen Jauntre. Healthy Lipophilic Fiber from Cactus. Monia Ennouri. Beneficial Effect Of Opuntia Ficus Indica Seeds And Oil On Animal Health. Saida Ncibi, Dalel Brahim, Amira Akacha, Mohamed Naceur Krifi, Sami Souid and Lazhar Zourgui. Biochemical and Immunolgical Toxicity of Chlorpyrifos in Mice. Changping Zou. Chemoprevention. Cactus Pear: A Natural Product In Cancer 47 49 51 57 61 63 2 3 3 18 29 33 36

42

Maria A. Livrea. Antioxidant activity of cactus pear fruits and bioavailability of betalain components in healthy humans. Abderrahman Ait Hamou. An Overview On Medicinal And Cosmetic Uses Of Cactus In Morocco. Hind Hammouch, Laila Bennghmouch, Abdellah Srhiri, Najat Hajjaji. Inhibition Of Iron Corrosion Using Opuntia Ficus Indica Extract.

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Editorial
Cactus, plant of the desert and flag of the poor, is increasingly attracting scientists and developers over the entire world. Just few years ago, no one would have imagined that outstanding advanced research laboratories in Germany, Italy, France, USA, South Korea, China, Mexico, and others would be investigating the potential uses of cacti as medicinal plants. In fact, Cactus was largely ignored by the scientific community until the beginning of 1980, when a multiplication of research and symposia occurred, resulting in a large number of publications. This renewed interest is to be ascribed in part to the multifunctionality of cactus fruits, pads and flowers. Also, in recent years a global trend is witnessed toward the use of natural phytochemicals present in various natural resources, including fruit, vegetables, oilseeds, and herbs, as antioxidants and functional foods. Natural antioxidants can be used in food industry, and there is evidence that they may exert their antioxidant properties within the human body. This issue of Cactusnet newsletter is dedicated to the coverage of medicinal uses of cactus products. While most of people are familiar with the use of cactus for fruit and fodder production few are aware of its potential usefulness as medicinal plant. Native Americans and ancient medicine have realized its anti-diabetic and anti-inflammatory properties, but Opuntia spp. have hardly been considered for the development of health-promoting food, most probably due to the little information available. The main objective of this issue therefore is to fill up this knowledge gap and to keep scientists, developers, and consumers abreast of recent advances made in the use of cactus cladodes, fruits and flowers as neutraceutical and medicinal resources. To confirm the importance given to this new area, a new research group is added to strengthen the structure of the International Technical Cooperation Network on Cactus (Cactusnet). Dr Monica Nazareno, Professor of organic chemistry at the University of Santiago del Estero, Argentina and co-editor of this issue, has been recently appointed to coordinate this working group. Created under the initiative of FAO, the cactus network now is fifteen years old, and has been attracting more and more people with increasing popularity due to the dedication and continuous support of Dr. Enrique Arias of FAO. The recent Sixth International Congress held during October 2007 in Brazil, has brought together more than 500 participants, with 34 invited papers and 234 other contributions, which testifies to the success and good health of the network. ICARDA serves the non-tropical dry areas where cacti play an important role as a drought tolerant plant resource for fruit, fodder and land conservation. Therefore, ICARDA is paying more and more importance to this crop, working closely with FAO to promote cactus in different parts of the world. The editors are especially grateful to FAO and ICARDA for their financial and technical support, and express their thanks and recognition to all partners, with a special mention of the many authors for their valuable contributions to this issue of Cactusnet newsletter. The editors
Monica Nazareno and Ali Nefzaoui

Scientific and Technical Contributions

Review Of Medicinal Uses Of Cactus
A. Nefzaoui1, Monica Nazareno2, and M. El Mourid3
(1) ICARDA/NARP, General Coordinator of FAO-CACTUSNET. ICARDA North Africa Regional Program. 1, Rue des Oliviers, El Menzah V, Tunis. Tunisia. A.nefzaoui@cgiar.org (2) Professor of Chemistry, Universidad Nacional Santiago del Estero, Argentina. manazar2004@yahoo.com (3) Regional Coordinator of ICARDA North Africa Regional Program, 1, Rue des Oliviers, El Menzah V, Tunis. Tunisia. M.elmourid@cgiar.org

Summary
Cactus is a xerophytic plant and an emergent crop, which evolved commercially during the second half of the XX century. The plant probably originated in Central Mexico, the region presenting the widest germplasm variability as well as the larger number of uses. Cactus species have developed phenological, physiological and a structural adaptation favourable to their development in arid environments, in which water is the main factor limiting the development of most plant species. Among these adaptations stand out its asynchronous reproduction, and its CAM metabolism, which combined with structural adaptations such as succulence allow this plant to continue the assimilation of carbon dioxide during long periods of drought and in this way reach acceptable productivity levels even in years of severe drought. Cactus cladodes, fruits, and flowers were traditionally used as medicines in several countries, and particularly in Latin America. Recent scientific investigations showed that cactus products may be efficiently used as a source of foods additives, mainly fibre, red dye and mucilages. Tablets, cookies and other forms of fibre derived from cactus cladodes are currently marketed in several American countries. Opuntia ficus-indica cladodes were directly administrated in tests to hypercholesterolemic rats. It was found a decrease of their blood plasma of cholesterol, HDL, LDL and triglycerides. It was also noted an inhibition of carragenan-induced oedema, a protective action against ethanol-induced ulcer, as well as other favourable effects. Similar tests were carried out by employing, instead of cladodes, mucilages and pectins extracted from cladodes, it was demonstrated that the above mentioned effects may be attributed to mucilages, and not significantly

pectins. It is also reported that the consumption of young cladodes or nopalitos reduces significantly obesity and blood glycaemia. Infusion of cactus flowers petals prevents prostates cancer. The use of cactus products as cosmetics is even more developed than medicinal applications. The most world wild known red dye or carmenic acid produced from cochineal growing on cactus cladodes are recognized by WHO as hypoallergenic dye thoroughly used in agro-industries and for cosmetics as lipsticks and powder. Shampoo, shower gels, and other forms of cosmetics are manufactured and marketed in several European and American countries.

1. Introduction
As a CAM plant, Opuntia spp. are characterized by a high water use efficiency of 410 mmol CO2 per mol H2O compared to C3- and C4-plants with 1.01.5 mmol and 23 mmol CO2 per mol H2O, respectively. Through succulence, the ability to store considerable quantities of water, the plant may survive despite harsh environmental conditions (Nobel and De la Barrera, 2003). Furthermore, and according to the same author, Opuntia exhibits the highest production rate of aboveground-growing plants. Interestingly, the biomass production was even found to increase upon otherwise deleterious rise of atmospheric CO2 concentrations (Nobel et al., 2002), thus counteracting the greenhouse effect. Cactus was largely ignored by the scientific world until the beginning of 1980, when there was a multiplication of research and symposia, resulting in a large number of publications. This renewed interest is to be ascribed in part to the multi-functionality of cactus fruits, pads

and flowers. Recent data has, in fact, revealed the high content of some chemical constituents, which can give added value to cactus products. Additionally, some of the constituents show promising characteristics in terms of functionality. In recent years there has been a global trend toward the use of natural phytochemicals present in natural resources, such as fruit, vegetables, oilseeds, and herbs, as antioxidants and functional foods. Natural antioxidant can be used in the food industry, and there is evidence that these substances may exert their antioxidant effects within human body. Cactus has a global distribution and is an important nutrient and food source. The growing demand for neutraceuticals is paralleled by an increased effort in developing natural products for the prevention or treatment of human diseases. According to several studies demonstrating both cactus fruit and cladode yielding high values of important nutrients, such as betalains, amino compounds including taurine, minerals, vitamins, as well as further antioxidants, the cactus pear (Opuntia spp.) appears to be an excellent candidate for the inclusion in food. Even though Native Americans and ancient medicine have realized its antidiabetic and anti-inflammatory function, Opuntia spp. have hardly been considered in the development of health-promoting food, most probably due to the scattered information available. 2. Functional compounds of cacti Recent shift toward natural products as food, with specific consumers preferences as low contents in calories, fats, and cholesterol and rich in fibres, minerals, vitamins, anti-oxidants and absence of artificial colours. This trend has led to the concept of functional foods which are a source of nutrients, but also good for the health. Opuntia species produce nutrients that fit with this context (Carmen Saenz, 2006). The great number of potentially active nutrients and their multifunctional properties make cactus pear (Opuntia spp.) fruits and cladodes perfect candidates for the production of healthpromoting food and food supplements. Although traditionally appreciated for its pharmacological properties by the Native Americans, cactus pear is still hardly recognized because of insufficient scientific information. However, recent studies on Opuntia spp. have demonstrated cactus pear fruit and vegetative cladodes to be excellent candidates for the development of healthy food.

Some constituents may be extracted and used as additives in food preparation or in the pharmaceutical and cosmetic sectors. Possible use of food colorants has been assessed. Betalains are found only in 10 families of the Caryophyllaleae. Betalains are nitrogenous chromalcaloids widely used in food industry. Pectins with enough galacturonic acid content are abundant in the fruit and can be used as food or cosmetic additive. Pectins in cactus fruit have a very low degree of methoxylation, suggesting that can be used ass gelling agent for low caloric food. Mucilages, which are complex polysaccharides, mainly composed of arabinose, galactose, rhamnose, and galacturonic acid (Lee et al., 1988) have a high water holding capacity, and so they can serve as thickening or emulsifying agents and form viscous or gelatinous colloids. Fibre, hydrocolloids (mucilages), pigments (betaines, carotenoides), minerals (calcium, phosphorus) are some of the functional compounds found in cactus cladodes and fruits. The analyses of malic and citric acids but also vitamin C may be helpful for nutritional and sensorial evaluation of the respective Opuntia sp. studied. Free and bound oxalic acids, but also rare compounds such as phorbic and piscidic acids could be valuable markers for specific species and their respective physiological stage. In addition, the presence and quantity of alkaloids in Opuntia spp. should be checked. In the light of recent reviews, promoting the knowledge of phenolic constituents in the everyday diet as well as their impacts on human health in-depth studies deserve special attention (Stintzing and Carle, 2005) Generally, cladodes are rich in pectin, mucilage and minerals, whereas the fruits are good sources of vitamins, amino acids and betalains. While the seed endosperm was reported to consist of arabinan-rich polysaccharides (Meyer et al., 1980), the principal seed coat component was D-xylan (Habibi et al., 2002). In addition to lipids, seeds have been reported to accumulate proanthocyanidins (Nieto, 1987). The fruit skin polysaccharide fraction has been subject to thorough investigations (Habibi et al., 20022003-2004), whereas the pectin substances in fruit pulp remain to be characterized. The flowers predominantly accumulate betalains and colorless phenolics (Salt et al., 1987).

Fibers
The healthy properties of fibres are well established: control of obesity, diabetes, and cholesterol (Sloan, 1994). There are two types of fibres: water soluble and insoluble. Soluble fibres include mucilage, gums, pectin, and hemi-celluloses. Soluble fibres are associated to the reduction of blood glucose and cholesterol and stabilize intestinal food transit. Insoluble fibres are composed mainly of cellulose, lignin, and a large portion of hemicelluloses. They are known for their capacity to retain water, favour ionic exchange, absorption of bile acids, minerals, vitamins and other interaction with microbes (Carmen Saenz, 2006). Dried cladodes powder contains about 43 % fibre, 28,5 % of which are insoluble. Some commercial products based on cladodes powder are already marketed, as Cactu fibra and Nutra Sweet in Mexico. These products are prepared with young cladodes (3-6 months), which have low content of insoluble fibre. This powder is also used to prepare several foods, as biscuits, creams, and other types of desserts (Saenz et al. 2002). Generally, about 18 % of cladodes powder is incorporated in such products.

natural dyes. Cactus fruits contains betalains (Sepulveda et al., 2003), which can be used in agri-food. Cactus betalains contains two types of pigments: betacyanins (red) and betaxantines (yellow). These pigments were used for yaourts (Idalsoaga, 2000) and orange juice (Moreno et al., 2003) preparations. Sixty one aromas have been found in cactus fruit flesh of Opuntia ficus-indica (Flath and Takahashi, 1978), most of them are alcohols. The most important aromas contributors in white, yellow and red Italian cultivars were (E,Z)-2,6-nonadien-1-ol and 2-methyl-acid methyl ester.

Anti-oxidants
Cactus fruits have a high anti-oxidant activity conferred by the presence of vitamin C, betacartenes, falvonides, and betalaines (Kuti, 2004; Galati et al., 2003). Fruits extract have an antioxidant activity of 4.2-5.3 mol Trolox f1 for white and yellow variety, respectively. This anti-oxidant activity is twice higher than pears, apples, tomato, bananas, white grapes, and similar to red grapes and grape fruit (Butera, 2002). Significant amounts of ascorbic acid are present in Opuntia ficus-indica, ranging from 180 to 300 mg/kg. Cactus fruit is higher in vitamin C than other common fruits, such ass apple, pear, grape, and banana. The most obvious feature of cactus pear fruits and flowers are the yellow (betaxanthins) and red (betacyanins) betalains, nitrogencontaining vacuolar pigments that replace anthocyanins in most plant families of the Caryophyllales including the Cactaceae (Zryd, 2004). Their identification in cactus pear fruit has been of renewed interest recently (Stintizing et al., 2003). In addition to color, the same pigments have shown antioxidant properties being higher than for ascorbic acid (Tesoriere et al., 2005). Lee et al. (2000) investigate the anti-oxidant activity of cactus cladodes and conclude that this anti-oxidant property is due to many compounds, particularly flavonoids (quercetin, myricetin), and vitamins. The presence of phenolics has been detected in cactus pulp fruit (Tesoriere et al., 2005b; Butera et al., 2002; Kuti, 2004). Kuti (1992) has reported an antioxidative effect due to the major flavonoids encountered in cactus fruits (quercetin, kaempferol and isorhamnetin). There is clear evidence that these compounds are more efficient antioxidants than vitamins, since phenolic compounds are able to delay prooxidative effects on proteins, DNA and lipids by the generation of stable radicals

Polysaccharides and Mucilage

Mucilage is a complex polysaccharide containing arabinose, galactose, xylose, galacturonic acid, and rhamnose. Because of its high ability to retain water, it is considered as a hydrocolloid, and can be used as thickener. Habibi et al. (2004) investigated fruit skin polysaccharides composition and find out they are composed of galactose and arabionse with 6.3:3.3 ratio, and classify them as arabinogalactane. Cactus mucilage may replace other thickeners as carboxymethylcellulose (Sepulveda et al., 2003). The aqueous ethanolic extract from the fresh cladodes of Opuntia dillenii Haw. showed potent radical scanvenging activity. Three new compounds, opuntioside I, 4-ethoxyl-6hydroxymethyl-alpha-pyrone, and kaempferol 7-O-beta-D-glucopyranosyl-(1-->4)-beta-Dglucopyranoside, were isolated from the extract (Qiu et al., 2002).

Pigments/dyes
WHO and EC regulations restrain the use of artificial dyes, favouring therefore the use of

(Shahidi et al., 1992). Furthermore, Opuntia ficus-indica polyphenolic compounds have been shown to induce a hyperpolarization of the plasma membrane and to raise the intracellular pool of calcium in human Jurkat Tcell lines (Aires et al., 2004). Flavonol derivatives detected in Opuntia ssp. have been recently compiled (Stintizing and Carle, 2005). When fruits are investigated, it must be taken into account that higher phenolic contents are expected in the peel, rather than the pulp. Consequently, from a nutritional point of view processing both peel and pulp appears to be advantageous. The fat soluble vitamin E or tocopherols, and beta-carotene are found in the lipid fraction of both the cactus fruit seed and pulp. The vitamin E homologues isoforms gamma- and delta-tocopherol are the main components in seed and pulp oils, respectively, amounting to about 80% of the total vitamin E content. Similar to beta-carotene, it is predominant in pulp lipids (Ramadan and Morsel, 2003). In Mexico, where cactus pads or nopalitos are commonly consumed as vegetable or cooked, it was reported that the above mentioned antioxidant are heat-resistant and remain active even after harvest is adequate storage is applied.

were reported (Hegwood, 1990). In Italy, a review on folk veterinary medicine compiles Opuntia ficus-indica as a plant species for ethnoveterinary (Viegi et al., 2003). The Aztecs used it for medicinal purposes as described in the Aztec Herbal of 1552. They referred to it as "nohpalli." Today the Mexicans still call the cactus plant "nopal. Parts of the prickly pear cactus have been used in various ways throughout the world. The Aztecs extracted the milky juice from the plant and mixed it with honey and egg yolk to provide an ointment to treat burns. The Chinese dressed abscesses with the fleshy pad of the plant. The Indians used the fruit for food and also made syrup from it to treat whooping cough and asthma. In Mexican traditional medicine, prickly pear cactus (nopal) is used for the treatment of diabetes and high cholesterol. Today nopal is a commonly called upon herbal agent for the treatment of Type 2 Diabetes by Mexican Americans as well as American Indians. The blood sugar- lowering action of nopal has been documented in a number of studies. Deep inside the African Kalahari desert, grows an ugly and bitter cactus called the Hoodia Gordonii. It thrives in extremely high temperatures, and takes years to mature. The San Bushmen of the Kalahari, one of the world's oldest and most primitive tribes, had been eating the Hoodia for thousands of years, to stave off hunger during long hunting trips. When South African scientists were routinely testing it, they discovered the plant contained a previously unknown molecule, which has since been christened P 57. The license was sold to a Cambridgeshire bio-pharmaceutical company, Phytopharm, who in turn sold the development and marketing rights to the giant Pfizer Corporation. In Italy, the flowers have served as a diuretic. A tea made from the blossoms has treated colitis. In Israel, researchers found that the dried flowers may be used to battle an enlarged prostate.

Lipids
Lipids are present in the peel, pulp, and seeds. Peel contains an appreciable amount of polyunsatured fatty acids, mainly linoleic acid, alpha-tocopherol, sterols, beta-carotene and vitamin K1 (Ramadan and Morsel, 2003a). The main sterol is b-sitosterol. Among amino acids present in the fruit, the highest is the nutraceutical taurine, with an amount of 572 mg/L (Stintzing et al., 2001).

3. Medicinal uses of cactus

Traditionally, cactus pads contributed considerably to the human diet in many countries, mainly in Latin American countries. They still serve as therapeutic agents and in folk medicine. Several cactus species, and mainly Opuntia fuliginosa and Opuntia streptacantha, have been used for the treatment of gastritis, fatigue, dyspnoe, and liver injury following alcohol abuse (Shapiro and Gong, 2002). Traditionally, heated poltices were applied to treat rheumatic disorders, erythemas and chronic skin infections, but also to improve digestion and enhance the general detoxification processes (Munoz et al., 1995). Recently, positive effects of cladodes on hyperglycemia, acidosis, and arteriosclerosis

3.1. Antioxidant capacity

The antioxidative action is one of many mechanisms by which fruit and vegetable substances might exert their beneficial health effects (Lee et al., 1999). The presence of several antioxidants (ascorbic acid, carotenoids, reduced glutathione, cysteine, taurine and flavonoids such as quercetin, kaempferol and isorhamnetin) has been detected in the fruits and vegetables of different varieties of cactus prickly pear (Lee et

al., 1999; Tesoriere et al., 2005; Kuti, 1992; Park et al., 1998). More recently, the antioxidant properties of the most frequent cactus pear betalains (betanin and indicaxanthin) have been revealed (Gentile et al., 2004; Tesoriere et al., 2003-2004-2005; Stinzing et al., 2005). Lee and Kim (2002) investigated Opuntia ficusindica var Saboten (OFS), widely spread cactus in Southwestern Korea, and where is used as a functional food. Antioxidant activity of OFS is reported to correspond to well-known antioxidants, such as catalase, -tocopherol, and ascorbic acid in a cell-free reactive oxygen species (ROS) generating system. It well established that oxidative stress may play a role in several diseases, such as heart disease, degenerative neuronal disease, and cancers. Many biochemical and clinical studies suggest that natural and synthetic antioxidant compounds are helpful in treating diseases mediated by oxidative stresses. Lee and Kim (2002) demonstrated that cactus cladodes have excellent antioxidant activities. On the other hand, numerous in vitro studies have demonstrated the beneficial effect of colorless phenolics and betalains (Stintizing et al., 2005; Dok-Go et al., 2003). These are generally attributed to the ability of antioxidants to neutralize reactive oxygen species such as singlet oxygen, hydrogen peroxide or H2O2, or suppression of the xanthine/xanthineoxidase system, all of which may induce oxidative injury, i.e. lipid peroxidation. Polyphenolics are antioxidants with well-known cardioprotective, anticancer, antiviral and antiallergenic properties (Carbo et al., 1999; Tapiero et al., 2002). Lee et al. (2002) demonstrated that antioxidative flavonoids, quercetin, (+)dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. Saboten have neuroprotective effects. They evaluated their protective effects against oxidative neuronal injuries induced in primary cultured rat cortical cells and their antioxidant activities. They found that quercetin inhibits H2O2- or xanthine (X)/xanthine oxidase (XO)-induced oxidative neuronal cell injury. Moreover, quercetin 3-methyl ether potently and dramatically inhibited H2O2 and X/XO-induced neuronal injuries. All above mentioned three principles markedly inhibited lipid peroxidation and scavenged 1,1-diphenyl-2-picrylhydrazyl free radicals. These results indicate that quercetin, (+)-dihydroquercetin, and quercetin 3-methyl ether are the active antioxidant principles in the fruits and cladodes of Opuntia

ficus-indica var. Saboten. Furthermore, quercetin 3-methyl ether appears to be the most potent neuroprotectant of the three flavonoids isolated from this plant. Sicilian cultivars of prickly pear (Opuntia ficusindica) produce yellow, red, and white fruits, due to the combination of two betalain pigments, the purple-red betanin and the yellow-orange indicaxanthin. The betalain distribution in the three cultivars and the antioxidant activities of methanolic extracts from edible pulp were investigated by Buetra et al. (2002). The yellow cultivar exhibited the highest amount of betalains, followed by the red and white ones. Indicaxanthin accounted for about 99% of betalains in the white fruit, while the ratio of betanin to indicaxanthin varied from 1:8 (w:w) in the yellow fruit to 2:1 (w:w) in the red one. Polyphenol pigments were negligible components only in the red fruit. When measured as 6-hydroxy-2,5,7,8tetramethylchroman-2-carboxylic acid (Trolox) equivalents per gram of pulp, the methanolic fruit extracts showed a marked antioxidant activity. Vitamin C did not account for more than 40% of the measured activity. In addition, the extracts dose-dependently inhibited the organic hydroperoxide-stimulated red cell membrane lipid oxidation, as well as the metaldependent and -independent low-density lipoprotein oxidation. The extract from the white fruit showed the highest protection in all models of lipid oxidation. These findings suggest that the above betalains contribute to the antioxidant activity of prickly pear fruits. It is appears that supplementation with cactus pear (Opuntia ficus-indica) fruit decreases oxidative stress in healthy humans: a comparative study with vitamin C. Short-term supplementation with prickly pear cactus fruit compared with vitamin C alone on total-body oxidative status in healthy humans was investigated by Butera et al. (2002). After supplementation with cactus pear fruit, malondialdehyde decreased by approximately 75%; GSH:GSSG shifted toward a higher value; and LDL hydroperoxides were reduced by almost one-half. Supplementation with vitamin C did not significantly affect any marker of oxidative stress. Authors concluded that consumption of cactus fruit positively affects the body's redox balance, decreases oxidative damage to lipids, and improves antioxidant status in healthy humans. Supplementation with vitamin C at a comparable dosage enhances overall antioxidant defence but does not significantly affect body oxidative stress. In the University of Messina, Italy, Galati and collaborators (2005) investigated the protective

effects of cactus fruit (Opuntia ficus-indica) juice against carbon tetrachloride (CCl4)induced hepatotoxicity in rats. The animals were treated orally with the juice (3 mL/rat) 2 h after administration of the hepatotoxic agent. Preventive effects were studied by giving the juice (3 mL/rat) for 9 consecutive days. On day 9 the rats received the hepatotoxic agent. Morphological and biochemical evaluations were carried out 24, 48 and 72 h after induction of the hepatic damage. Data show that O. ficus-indica fruit juice administration exerts protective and curative effects against the CCl4-induced degenerative process in rat liver. Histology evaluation revealed a normal hepatic parenchyma at 48 h; the injury was fully restored after 72 h. Moreover, a significant reduction in CCl4-induced increase of GOT and GPT plasma levels is evident; these data are in agreement with the functional improvement of hepatocytes. Authors assumed that cactus fruit juice contains many phenol compounds, ascorbic acid, betalains, betacyanins, and a flavonoid fraction, which consists mainly of rutin and isorhamnetin derivatives. Hepatoprotection may be related to the flavonoid fraction of the juice, but other compounds, such as vitamin C and betalains could, synergistically, counteract many degenerative processes by means of their antioxidant activity.

predominant sterol in cactacea (Salt et al., 1987). Numerous studies have evocated the analgesic and anti-inflammatory actions of the genus Opuntia by using either the fruit extract from Opuntia dillenii , the lyophilized cladodes (Galati et al., 2001), or the phytosterols from fruit and stem extracts (Park et al., 1998). Park et al. (2001) identified beta-sitosterol as the active anti-inflammatory principle from the stem extract. Gastric lesions in rat animal studies were reduced both by stem and fruit powders (Lee et al., 2001; 2002). Reduction of acute inflammation by ethanolic O. ficus-indica stem extracts was ascribed to a lower leucocytes migration. In contrast to nonsteroidal inflammation inhibitors, no adverse effects were noted. Therefore, cladode extracts were proposed for inflammation treatment. In further experiments with a methanolic extract from O. ficus-indica cladodes, the fractions obtained after reextraction with hexane and ethyl acetate were most efficient to accelerate the healing process (Clark and Parfitt, 1980). A follow-up study identified beta-sitosterol to be the active principle. An ethanolic extract of the cladodes (300600 mg/kg body weight) from O. ficus-indica var. saboten showed a similar analgesic effect as acetylsalicylic acid (200 mg/kg body weight) without toxic effects in mice (LD50 > 2 g/kg body weight) even at high dosages. The ethanol extracts of Opuntia ficus-indica fruits and inhibited the writhing syndrome induced by acetic acid, indicating that they contain analgesic effect (Park et al., 1998). The oral administrations of these extracts suppressed carrageenan-induced rat paw edema and also showed potent inhibition in the leukocyte migration of CMC-pouch model in rats. Moreover, the extracts suppressed the release of beta-glucuronidase, a lysosomal enzyme in rat neutrophils. It was also noted that the extracts showed the protective effect on gastric mucosal layers. From these results it is suggested that the cactus extracts contain anti-inflammatory action having protective effect against gastric lesions.

3.2. Anti-inflammatory and analgesic properties

Opuntia ficus-indica cladodes contain a polysaccharide fraction that can retain water. The cladodes are used in Sicilian folk medicine as cicatrizant. Enza Maria Galati and collaborators (2003) from the University of Messina, Italy, tested a wound-healing topical preparation containing 15% cladode extract. A fast regeneration of the tissue was ascribed to inflammation inhibition, stimulation of the fibroblast migration with accelerated collagen formation and faster angiogenesis. Evidently, the Opuntia ficus-indica treatment accelerates wound healing, probably by involving the proliferation and migration of the keratinocytes in the healing process. In an earlier work, the same authors demonstrated in 2000 the anti-inflammatory activity of Opuntia ficus-indica cladodes, and the correlation between the two phenomena is possible. Also, the angiogenesis is an essential process in wound healing (Brown et al., 1992), and these activators could be a low-molecularweight component of the cladodes (Lee et al., 1998) as monosaccharide residues, polyphenols or B-sitosterol, that is the

3.3. Antiulcerogenic effect and healing properties

Opuntia ficus-indica cladodes are used in traditional medicine of many countries for their cicatrisant activity. The major components of cladodes are carbohydrate-containing polymers, which consist of a mixture of

mucilage and pectin. In Sicily folk medicine Opuntia ficus-indica (L.) Mill. cladodes are used for the treatment of gastric ulcer. Galati et al. (2002) studied the effect of administration of lyophilized cactus cladodes on experimental ethanol-induced ulcer in rat. The ultrastructural changes were monitored by transmission electronic microscopy (TEM) confirming the protective effect induced by administration of lyophilized cladodes, and revealed a protective action against ethanol-induced ulcer. This protective effect is probably due to the mucilage. Indeed, the major components of cladodes are carbohydrate-containing polymers, which consist of a mixture of mucilage and pectin. The OFI cladodes administration gives rise to cytoprotection phenomena by breaking up the epithelial cells and stimulating an increase in mucus production. When OFI cladodes are administered as a preventive therapy, keep the gastric mucosa under normal condition by preventing mucus dissolution caused by ethanol and favouring mucus production. An increase of mucus production is also observed during the course of the curative treatment. The treatment with OFI cladodes provokes an increase in the number of secretory cells. Probably, the gastric fibroblasts are involved in the antiulcer activity (Galati et al., 2002) In another work, Galati et al. (2003) investigated the cactus fruit juice contents of ascorbic acid, total polyphenols, and flavonoids. In the juice, ferulic acid was the chief derivative of hydroxycinnamic acid and the mean concentration of total phenolic compounds was 746 microg/mL. The flavonoid fraction, analyzed by high-performance liquid chromatography-diode array detection, consisted of rutin and isorhamnetin derivatives. The juice showed antioxidant activity, probably due to the phenolic compounds that are effective radical scavengers. The preventive administration of the juice inhibited the ulcerogenic activity of ethanol in rat. Light microscopy observations showed an increase in mucus production and the restoration of the normal mucosal architecture. Additionally, Lee et al. (2001, 2002) postulated an antiulcerogenic effect of the cladode or fruit powder from O. ficus-indica var. saboten Makino. Stomach lesions triggered by hydrochloric acid/ethanol or hydrochloric acid/acetylsalicylic acid were reduced, but no anti-inflammatory effect could be proven. The secretion rate of both gastric juice as well as the pH value remained constant, which has been confirmed by Galati et al. (2001). However, the protective effect was ascribed to

the cladodes hydrocolloid acting as a buffer, spreading out on the gastric mucosa and increasing mucus production by enhancing the number of secretory cells.

3.4. Hypoglycemic and antidiabetic effects

The cactus cladodes have been used traditionally to treat diabetes in Mexico (Dominguez Lopez, 1995). Nowadays, Opuntia spp. is amongst the majority of products recommended by Italian herbalists that may be efficacious in reducing glycemia (Cicero et al., 2004). Human studies in the 80es demonstrated glucose and insulin levels in healthy fasting subjects were stable when eating cladodes. The positive contribution to overall health in diabetes mellitus type II (noninsulin-dependent diabetes) patients was assumed to be due to reduced postprandial sugar absorption. Following a glucose challenge test, the increase in insulin and glucose were retarded. Also, the glucose and insulin plasma levels were reduced. After 10 days of cladode ingestion prior to meals, a significant reduction of the serum glucose level was noticed [Meckes-Lazyoa and Roman Ramos, 1986). Since these effects did not depend on glucagon, cortisone, and human growth hormone levels, which are closely interrelated with glucose metabolism, a gastric enterohormone was held responsible for the hypoglycemic effect (Munoz de Chavez, 1995). To assess the hypoglycemic effect of the Opuntia streptacantha Lemaire cladodes, FratiMunari et al. (1988) conducted an experiment with patients with non-insulin-dependent diabetes mellitus. After the intake of O. streptacantha Lem., serum glucose and serum insulin levels decreased significantly. Authors did not identify the mechanism responsible of this action, but suggest increased insulin sensitivity. Wolfram et al. (2002) reached quite similar conclusions. Indeed, they demonstrate that cladodes have an anti-hyperglycemic effect that was proposed as alternative to oral antidiabetics, thereby preventing insulin resistance. The hypoglycaemic effect seemed to be related to improved sensibility of the pancreatic cells with a concomitant improved glucose usage (Wolfram et al., 2002). These effects deserve special attention since the populations of developed countries are increasingly suffering from obesity and diabetes symptoms urgently requiring effective countermeasures. The efficiency of cactus cladodes in lowering blood sugar was observed in both fried and

raw materials. Interspecific variations of the plant material, harvest time, and application modes (oral, intravenous) should be considered. Some studies have demonstrated the hypoglycemic activity of the prickly pear cactus extract on non-diabetics and diabetic-induced rats or diabetic humans (Enigbokan et al., 1996). In a study on rats, the combination of insulin and purified extract of cactus (Opuntia fuliginosa Griffiths) was found to reduce blood glucose and glycated hemoglobin levels to normal (Enigbokan et al., 1996). In this study, the oral dose of extract (1 mg/kg body weight per day) necessary to control diabetes contrast with the high quantities of insulin required for an equivalent hypoglycemic effect. A recent study has shown that a supplementation of rats diets with cactus seed oil (25 mg/kg) decreases the serum glucose concentration, which is associated with a glycogen formation in the liver and skeletal muscle (Laurenz et al., 2003). These observations were explained by a potential induction of insulin secretion, converting glucose to glycogen. Wolfram and collaborators (2003) from the University of Vienna, Austria, suspected a soluble fibre in lowering cholesterol levels in both animals and man with hyperlipidemia as well as being able to slow carbohydrate absorption and hence reduce the postprandial rise in blood glucose and serum insulin in patients with type-II diabetes. The aim of their work was to investigate the effect of cactus consumption on glucose and lipid metabolism. Cactus consumption leads to a decrease of total cholesterol (12%), low-density lipoproteincholesterol (15%), apolipoprotein B (9%), triglycerides (12%), fibrinogen (11%), blood glucose (11%), insulin (11%) and uric acid (10%), while body weight, high-density lipoprotein-cholesterol, apolipoprotein A-I, and lipoprotein(a) remained unchanged. Authors conclude that the hypocholesterolemic action of cactus may be partly explained by the fibre (pectin) content, but the hypoglycaemic actions (improvement of insulin sensitivity) in the nonobese, non-diabetic need further investigation to get more insights on the potential advantage of treating the metabolic syndrome. An amazing effect of cacti in controlling appetite has been recently reported in several media and it may change the life of people suffering from obesity. It is perceived as a new miracle diet ingredient. Imagine an organic pill that kills the appetite and attacks obesity. It has no known side-effects, and contains a molecule that fools your brain into believing

you are full. This pill is already sold through internet and it is manufactured from a cactus ugly plant with a bitter taste named Hoodia gordonii. This unattractive plant which sprouts about 10 tentacles, and is the size of a long cucumber is growing in the Kalahai desert. Each tentacle is covered in spikes which need to be carefully peeled. Inside is a slightly unpleasant-tasting, fleshy plant. The plant is said to have a feel-good almost aphrodisiac quality. The San bushmen have eaten this plant for years, which help to suffer hunger and starvation when hunting. Trials were conducted on animals and human. When offered Hoodia plant, rats, a species that will eat literally anything, stopped eating completely. Trials on human with two groups, one half were given Hoodia, and the other half placebo. Fifteen days later, the Hoodia group had reduced their calorie intake by 1000 a day. ."What the Hoodia seems to contain is a molecule that is about 10,000 times as active as glucose. "It goes to the mid-brain and actually makes those nerve cells fire as if you were full. But you have not eaten. Nor do you want to. When South African scientists were routinely testing the plant, they discover that it contains unknown molecule, which has since been named P57. The licence was sold to a Cambridgeshire bio-pharmaceutical company, Phytopharm, who in turn sold the development and marketing rights to the giant Pfizer Corporation.

3.5. Anti-hyperlipidemic and cholesterol lowering effect

Experimental evidence suggests that cactus cladodes reduces cholesterol levels in human blood and modify low density lipoprotein (LDL) composition (Fernandez et al., 1992; Frati, 1992). Galati et al. (2003) have found that the cholesterol, LDL and triglyceride plasma levels of rats were strongly reduced after 30 days of a daily administration (1 g/kg) of lyophilized cladodes of Opuntia ficus-indica L. Mill. This effect is generally attributed to the high fiber content of the cladodes, although other active ingredients (such as betacarotenes, vitamin E and beta-sitosterol) may be involved. Oil from cactus seeds fed to rats (25 g/kg) decreases plasma total cholesterol and LDL (VLDL) cholesterol with no effect on HDL-cholesterol concentrations ( Ennouri et al., 2005). The use of cactus may also lower blood lipid levels. In a recent study, the daily consumption of 250 grams of cooked cactus cladodes lowered total cholesterol and LDL

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cholesterol levels (but not HDL cholesterol or triglyceride levels) in 15 young patients with familial hypercholesterolemia. Although there are many species of Opuntia, few varieties have been positively shown to be effective in normalizing blood sugar or blood lipid levels (Feugang, 2006). The anti-hyperlipidemic effect after cladode ingestion was investigated only in many studies (i.e. Wolfram et al., 2002-2003; Budinski). In general, a prolonged period of satiety was registered after cladode consumption. In a series of studies with Guinea pigs, Fernandez et al. (1990, 1994) demonstrated that the reduction of blood lipids triggered by isolated pectin from Opuntia was due to the enhanced binding of bile acid. It was concluded that through reduced bile absorption in the colon the enterohepatic circle was disrupted (Fernandez et al., 1990). In a follow-up study, the same authors presented evidence that the low-density lipoprotein (LDL)-catabolism was considered to be more important than the modulation and de novo synthesis in the liver. The same pectic-like substances were held responsible for decreased lipid absorption, lower blood lipid levels, and finally weight reduction. In a recent study it was shown that a daily consumption of 250 g of prickly pear pulp reduced the risk of thrombosis in patients suffering from hyperlipidemia and diabetes (Wolfram et al., 2003). Wolfram et al. (2002) reported a reduction of total cholesterol, LDL, apolipoprotein levels, triglycerides, fibrinogen, blood glucose, insulin and urate, while body weight, high-density lipoprotein (HDL)cholesterol, apolipoprotein A-1 and lipoproteinA levels were found to remain unchanged. The anti-hyperlipidemic effects were ascribed to the pulp pectin, which both reduced lipid absorption and increased fecal sterol excretion, thus disrupting the enterohepatic circle.

detected in the serum after administration of an Opuntia megacantha extract. These results show cactus potential for gout treatment as claimed by Bwititi et al. (1997). In this context, oxalic acid needs to be taken into consideration since calcium availability was found to be decreased due to sequestration by oxalic acid and high oxalic acid contents resulted in an enhanced urinary calcium oxalate excretion (Nerd et al., 1997). After Opuntia extract ingestion, the water intake rose and the urine volume was increased (Bwititi et al., 1997). Galati et al. (2002) investigated the diuretic effect of a 15% extract from flowers, fruits, and peeled cladodes from Opuntia ficus-indica, respectively. The latter showed the highest diuretic effect while urea levels in blood and urine remained unchanged. The diuretic effect was chiefly ascribed to the high potassium content of Opuntia cladodes amounting to 548 mg/kg.

3.7. Anti-cancer effect

It has been noted that Native Americans have a lower cancer rate when compared to white and African Americans (Cancer statistics, 2004). Both cactus fruits and cladodes which contain multiple antioxidants have been used as a dietary supplement for centuries by Native Americans. The goal of cancer prevention is to delay or block the processes of initiation and progression from pre-cancerous cells into cancer. Cancer chemoprevention, which targets normal and high risk populations, involves the use of drugs or other chemical agents to inhibit, delay, or reverse cancer development (Kelloff et al., 1999). Medical benefits from plant forms have been recognized for centuries. Herbs have been used in Chinese medicine for thousands of years to cure diseases and heal wounds. Recently, it has been found that components in green tea and grape seeds have anticancer effects (Kim et al., 2004). Also, as a rule, herbs and natural products lack much of the toxicity that is present in synthetic chemicals, thus, enhancing their appeal for long term preventive strategies. Although hundreds of agents have been developed in the United States during the past decade, only a few new drugs have actually been approved (Kelloff et al., 2000). The development of chemopreventive agents is slow and inefficient. More effective and less toxic agents, including natural products, are needed if we are to reach the goal of cancer

3.6. Diuretic effect

An ethanolic extract from Opuntia megacantha was reported to decrease blood glucose but also affect kidney function in rats (Bwititi et al., 2000). While sodium excretion was enhanced, potassium levels in urine decreased. In contrast, sodium, calcium, and magnesium levels in the plasma dropped, but phosphate, creatinine, and urea concentrations increased (Bwititi et al., 2000). Both observations were related to the hormonal regulatory mechanisms of the kidney. In rats, uric acid excretion was enhanced and lower uric acid levels were

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prevention, both primary and secondary. The development of effective and safe agents for prevention and treatment of cancer remains slow, inefficient, and costly. The key to effective chemoprevention is the identification of a chemopreventive agent(s) that can effectively inhibit cancer development without toxic side effects. In an Italian 4-HPR trial, retinoids showed the preventive effect on ovarian cancer only during the period while the drug was taken. After cessation of treatment, the incidence of ovarian cancer increased to the level that was observed in the untreated control group (Veronesi, 1999). Therefore, chemopreventive agents may need to be used for a long period of time to be effective. As a result, identification of agents with little or no toxicity becomes important. Recent studies suggests that the cactus fruit extract (i) inhibits the proliferation of cervical, ovarian and bladder cancer cell lines in vitro, and (ii) suppresses tumor growth in the nude mice ovarian cancer model in vivo (Zou et al., 2005). The intra-peritoneal administration of cactus extract solution into mice did not affect the animal body weight, which indicated that cactus did not have a significant toxic effect in animals. More importantly, tumor growth inhibition was comparable to the synthetic retinoid N-(4-hydroxyphernyl) retinamide (4HPR), which is currently used as a chemopreventive agent in ovarian cancer chemoprevention. The mechanism of action as well as the component(s) by which cactus pear extract exerts these effects is not yet elucidated. However, an extrinsic effect through an activation of membrane death receptors and intrinsic actions via the mitochondria, playing a pivotal role by releasing a number of molecules favorable to the induction of apoptosis, are suspected. Further investigations are needed to identify active components and the respective underlying mechanisms Zou et al. (2005) showed that growth inhibition of cultured-cancer cells was associated with an increase in apoptotic cells and the cell cycle arrest at the G1-phase. Moreover, the inducedgrowth inhibition seems dependent on the P53 pathway, which is the major tumor suppressor. Annexin IV was increased and the VEGF decreased in the tumor tissue obtained from animals having received the cactus solution. The same authors confirm that cactus fruit extract inhibited growth of different cancer cells in vitro and in vivo. Cactus products inhibited cancer cell growth with concentrations as low as 5%; cell cycle was also affected at this

concentration with an increase in G1 phase. However, apoptosis was observed at a higher concentration of 10% and 25%. They have also compared cactus with the chemopreventive agent 4-HPR in nude mice. Both cactus and 4-HPR inhibited ovarian cancer growth. The anti-carcinogenic properties of natural and synthetic retinoids have been suggested to be due, in part, to the antioxidant effect, increased consumption of fruit and vegetables is associated with prevention of various human diseases, and the oxidative damage is an important etiologic risk factor for many diseases, including cancer and heart disease. Carcinogenesis may be viewed as a process of progressive disorganization. This process is characterized by the accumulation of genotypic changes and corresponding tissue and cellular abnormalities including loss of proliferation and apoptosis controls. A dietary agent that can increase anti-proliferation pathways and change cell cycle in cancer cells without toxicity would be a potential agent for chemoprevention. Although the mechanism for cactus pear extract in cancer prevention is unclear, the study conducted by Zou et al. (2005) shows that cactus fruit extracts alter the expression of certain genes related to cell growth and apoptosis. Quercetin is one of the components of cactus pear extracts. Results of Zou et al. (2005) and Hansen et al. (1997) suggest quercetin might be one of the active compounds responsible for the anticarcinogenetic and apoptosis-induction effects of cactus pear extracts. In our study, cactus pear extracts decreased VEGF expression, suggesting that cactus pear extracts might have inhibitory effects on angiogenesis, an important factor contributing to tumor growth and metastasis. It can be concluded as stated by Zou et al. (2005) that cactus pear extracts could be a candidate in cancer prevention for both normal and high-risk populations and prevention of recurrence in patients with previous cancers. This product holds promise for long-term use because of the safety of food-derived products and the fact that they are not perceived as a "chemical", and that mechanisms involved need to be further investigated.

3.8. Antiviral effect

A cladode extract from Opuntia streptacantha Lem. was reported to exhibit antiviral properties towards DNA viruses, such as herpes, and RNA viruses, such as influenza type A and human immunodeficiency virus (HIV)- 1. The active principle was located in

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the outer noncuticular tissue and ascribed to a protein with unknown mechanisms of action (Ahmad et al., 1996). Both the replication of DNA and RNA viruses was inhibited while the extract from the parenchyma acted both preventively and post-infectionary. In security tests on mice, horses (27 g/day over the period of 24 weeks) and finally humans (6 g/day for 1 month or 3 g/d over 6 months), all dosages were well tolerated (Ahmad et al., 1996). An interesting study by Ahmad et al. (1996) demonstrated that administration of a cactus stem extract (Opuntia streptacantha) to mice, horses, and humans inhibits intracellular replication of a number of DNA- and RNAviruses such as Herpes simplex virus Type 2, Equine herpes virus, pseudorabies virus, influenza virus, respiratory syncitial disease virus and HIV-1. An inactivation of extracellular viruses was also reported by the same authors. However, the active inhibitory component(s) of the cactus extract used in this study was not investigated, and as of yet, no further study dealt with this specific topic.

may prove beneficial in BPH treatment. Bhattacharya (2004) reported that cactus extract offers hangover help. The purpose was not to eliminate the symptoms of the alcohol hangover so people could drink with impunity. The only known cure for the alcohol hangover is to not drink. Scientists interest was to discover what is causing the alcohol hangover, and to investigate what harms may be associated with the syndrome. Hangovers, or veisalgia as the condition is called medically, carry huge economic and health consequences. But despite this, little is known about its mechanism. It has been associated with a heightened inflammatory response by the body to alcohol impurities called congeners and some preservatives. Extract from the skin of Opuntia ficus-indica fruit had previously been shown to dampen inflammatory response. The overall assessment of hangover severity, based on nine symptoms, was reduced by nearly a fifth, but this result was not statistically significant. The researchers also measured levels of a protein produced by the liver, called C-reactive protein, which is thought to be involved in the inflammation process. The higher the levels, the worse the hangover, they found. This is the first study to show this, the team believes. Levels of this protein were also 40 per cent higher in the people who took placebo pills compared with those who took the Opuntia ficus-indica. The researchers therefore believe that Opuntia ficus-indica eases hangovers by soothing the inflammatory response to alcohol. A capsule is already marketed now days and could help for hangovers. Recent work (Lee et al., 2006) showed that Opuntia ficus-indica may have a neuroprpotective activity. Indeed, activated microglia by neuronal injury or inflammatory stimulation overproduce nitric oxide (NO) by inducible nitric oxide synthase (iNOS) and reactive oxygen species (ROS) such as superoxide anion, resulting in neurodegenerative diseases. The toxic peroxynitrite (ONOO-), the reaction product of NO and superoxide anion further contributes to oxidative neurotoxicity. A butanol fraction obtained from 50% ethanol extracts of Opuntia ficus-indica var. Saboten (Cactaceae) stem and its hydrolysis product inhibited the production of NO in LPS-activated microglia in a dose dependent manner (IC50 15.9, 4.2 g/ml, respectively). They also suppressed the expression of protein and mRNA of iNOS in LPS-activated microglial cells at higher than 30 g/ml as observed by western blot analysis and RT-PCR experiment. Moreover, they

3.9. Other positive health effects

Cactus fruits, cladodes or flower infusions have been traditionally used as folk medicine to treat other ailments such as ulcers, allergies, fatigue and rheumatism, to alleviate alcoholic hangover, and as an antiuric and diuretic agent. Amongst the flavonoids extracted from either the cactus stem or fruit, quercetin 3methyl appears to be the most potent neuroprotector (Dok-Go et al., 2003). The cactus flower extract was able to exert an effect on benign prostatic hyperplasia (BPH) through the inhibition of aromatase and 5a reductase activities, both of which are involved in androgen aromatization and testosterone reduction (Jonas et al., 1998). The investigation conducted by Jonas et al. (1998) evaluated the ability of cactus flower extracts to exert an effect on BPH through possible inhibition of such processes as lipid peroxidation, androgen aromatization and testosterone reduction. Cactus flower extracts indeed inhibited aromatase and 5alpha reductase activity in cultured foreskin fibroblasts, and also in human placental and prostatic homogenates. The inhibitory activity in both instances was associated with the dichloromethane or ethanol (methanol) extracts, while a marked antioxidative activity was associated with the aqueous extract. The finding that cactus flower extracts interfere concurrently in vitro with aromatase and reductase activity as well as with free radical processes suggests that these substances

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showed strong activity of peroxynitrite scavenging in a cell free bioassay system. These results imply that Opuntia ficus-indica may have neuroprotective activity through the inhibition of NO production by activated microglial cells and peroxynitrite scavenging activity.

under-stood. From the presented data, it appears that Opuntia spp. has been subject to intensive exploitation due to its great compositional diversity. Nowadays, this hidden knowledge needs to be discovered and re-evaluated. Sophisticated analytical approaches and innovative processing technologies will open new avenues to further promote the use of cactus pear stems, fruits and flowers in food, medicine, cosmetic, and pharmaceutical industries. An increasing demand would help encourage farmers to increase their acreage and thus also help to counterbalance erosion and increasing atmospheric CO2 levels. Although much research still needs to be done, concerted actions of taxonomists, plant breeders, agriculturists, food technologists, nutritionists and pharmacologists will help discover and understand the big potential of the Opuntia cactus. The exact botanical classification of the respective Opuntia spp. under investigation and the growing location and time of harvest are prerequisites for analytical and pharmacological studies. The exact plant parts used in the extraction and processing conditions need to be accurately documented to allow proper data evaluation.

4. Conclusions
Not only the biofunctional properties but also the assumed roles of cladode polysaccharides in the medicinal sector are interesting such as cholesterol reduction and its preventive action in diabetes and adipositas therapies. Since the hydrocolloid fraction is also rich in proteins, further research should be conducted whether the alleged positive effects are due to the pectic substances or rather to the protein fraction. Furthermore, it should be investigated whether these proteins are acting as heatshock and anti-freeze proteins [213220] in the cactus tissue. Whether the same proteins acting as repair molecules in plants could be used for pharmaceutical purposes merits further consideration. Special attention should also be given to the oligosaccharide fraction that has not been characterized so far. Immunomodulatory properties have been postulated for glucans from roots of Periandra mediterranea (Vell.) Taub. and glucomannans from Aloe sp. According to recent publication, small oligosaccharides with a polymerization degree of 6 and above with a tendency to form helical structures are potent immune modulators. In summary, the constituents of Opuntia cladodes are only partly known and often not quantitatively determined. Investigations were mostly performed 20 years ago and need to be validated with up-to-date methods. Furthermore, data stem from all kinds of different Opuntia spp. and it is open to question whether the botanical classification has properly been assessed in each case. Beside a sound systematic classification, the background of cladode physiology needs to be considered. Based on additional data, a reliable nutritional evaluation can be performed. The technofunctional properties of the respective extracts can be efficiently exploited for manifold food, cosmetic and medicinal applications. Analogies to the multiple uses of Aloe sp. are obvious. Whether isolated substances or rather a concerted action of several components in the complex plant matrix are responsible for the big variety of biological activities remains unknown, let alone the underlying mechanisms of the traditional curative treatments being still little

5. References
Ahmad A., J. Davies, S. Randall & G.R.B. Skinner,1996, Antiviral properties of extract of Opuntia streptacantha. Antiviral Research 30, 75-85. Aires V., S. Adote, A. Hichami, K. Moutairou, E. S. E. Boustani & N. A. Khan, 2004. Modulation of intracellular calcium concentrations and T cell activation by prickly pear polyphenols. Mol Cell Biochem 260, 103-110. Butera D., L. Tesoriere, F. Di Gaudio, A. Bongiorno, M. Allegra, A.M. Pintaudi, R. Kohen & M.A. Livrea, 2002. Antioxidant activities of Sicilian prickly pear (Opuntia ficus indica) fruit extracts and reducing properties of its betalains: Betanin and indicaxanthin. J Agric Food Chem 50, 68956901. Bwititi, P., Musabayane, C. T., Nhachi, C. F. B., 2000. Effects of Opuntia megacantha on blood glucose and kidney function in streptozotocin diabetic rats. J. Ethnopharmacol., 69, 247 252. Bwititi, P., Zamurawo, M., Mabhachi, G., Mashanga, N., 1997. Toxic and hypericaemic effects of Opuntia megacantha extract in rats. Phytother. Res., 11, 389391. Cancer statistics, 2004. American cancer society Carbo N., P. Costelli, F.M. Baccino, F.J. LopezSoriano & J.M. Argiles, 1999. Resveratrol, a natural product present in wine, decreases tumour growth in a rat tumour model. Biochem Biophys Res Commun 254, 739

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743. Cicero A.F.G., G. Derosa & A. Gaddi, 2004: What do herbalists suggest to diabetic patients in order to improve glycemic control? Evaluation of scientific evidence and potential risks. Acta Diabetologica 41, 91-98. Clark W.D. & B.D. Parfitt, 1980. Flower flavonoids of Opuntia series Opuntiae. Phytochem 19, 1856-1857. Dok-Go H., K.H. Lee, H.J. Kim, E.H. Lee, J. Lee, Y.S. Song, Y.-H. Lee, C. Jin, Y.S. Lee & J. Cho, 2003. Neuroprotective effects of antioxidative flavonoids, quercetin, (+)dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. saboten. Brain Res 965, 130-136. Dominguez Lopez, A., 1995. Review: use of the fruits and stems of the prickly pear cactus (Opuntia spp.) into human food. Food Science and Technology International, 1, 6574. Enigbokan, M. A., Felder, T. B., Thompson, J. O., Kuti, J. O., Ekpenyong, K. I., 1996. Hypoglycaemic effects of Opuntia ficus indica Mill., Opuntia lindheimeri Engelm. and Opuntia robusta Wendl. in streptozotocininduced diabetic rats. Phytother. Res., 10,379382. Ennouri M., E. Bourret, L. Mondolot & H. Attia, 2005. Fatty acid composition and rheological behaviour of prickly pear seed oils. Food Chem 93, 431-437. Fernandez M.L., E.C. Lin, A. Trejo & D.J. McNamara,1992. Prickly pear (Opuntia sp.) pectin reverses low density lipoprotein receptor suppression induced by a hypercholesterolemic diet in guinea pigs. J Nutr, 122, 2330 2340. Fernandez, L.M., Lin, E.C.K., Trejo, A., McNamara, D.J., 1994. Prockly pear (Opunita sp.) pectin alters hepatic cholesterol metabolism without affecting cholesterol absorption in Guinea pigs fed a hypercholesterolemic diet. J. Nutrition, 124, 817-824. Feugang, J.M., Konarski, P., Zou, D., Stintzing, F.C., and Zou, C. 2006. Nutritional and medicinal use of cactus (Opuntia spp.) cladodes and fruits. Frontiers in Bioscience 11, 2574-2589. Flath, R.A., and J.M. Takahashi, 1978. Volatile constituents of prickly pear (Opuntia ficus indica Mill.), de Castilla variety. Journal of Agriculture and Food Chemistry, 26, 835837. Frati A., 1992. Medical implication of prickly pear cactus. In: Proc. 3rd Annual Texas prickly pear council. Eds: Felkar P, Moss LR, 2425 July, Kingsville, Texas, 29-34. Galati E. M., M. R. Mondello, E. R. Lauriano, M. F. Taviano, M. Galluzzo & N. Miceli, 2005. Opuntia ficus indica (L.) Mill. Fruit Juice Protects Liver from Carbon Tetrachlorideinduced Injury. Phytother Res 19, 796800. Galati E.M., M.M. Tripodo, A. Trovato, A. d'Aquino &

M.T. Monforte, 2003. Biological activity of Opuntia ficus indica cladodes II: Effect on experimental hypercholesterolemia in rats. Pharm Biology 41, 3, 175-179. Galati E.M., M.M. Tripodo, A. Trovato, N. Miceli, & M.T. Monforte, 2002. Biological effects of Opuntia ficus indica (L.) Mill. (Cactaceae) waste matter. Note I: diuretic activity. J Ethnopharmacol 79, 17-21. Galati E.M., M.T. Monforte, M.M. Tripodo, A. dAquino & M.R. Mondello, 2001: Antiulcer activity of Opuntia ficus-indica (L.) Mill. (Cactaeceae): ultrastructural study. J Ethnopharmacol 76, 19. Galati, E. M., Mondello, M. R., Monforte, M. T., Galluzo, Gebhardt, R., 2003. Variable influence of kaempferol and myricetin on in vitro hepatocellular cholesterol biosynthesis. Planta Med., 69, 10711074. Gentile C., L. Tesoriere, M. Allegra, M.A. Livrea & P. DAlessio, 2004. Antioxidant betalains from catus pear (Opuntia ficus-indica) inhibit endothelial ICAM-1 expression. Ann. N. Y. Acad. Sci. 1028, 481486. Grindlay, D., Reynolds, T., 1986. The Aloe vera phenomenon: a review of the properties and modern uses of the leaf parenchymagel.J. Ethnopharmacol., 16, 117151. Habibi Y., A. Heyraud, M. Mahrouz & M.R. Vignon, 2004. Structural features of pectic polysaccharides from the skin of Opuntia ficus-indica prickly pear fruits. Carbohydr Res 339, 1119-1127. Habibi Y., M. Mahrouz & M.R. Vignon, 2002. Isolation and structure of D-xylans from pericarp seeds of Opuntia ficus indica prickly pear fruits. Carbohydr Res, 337, 1593-1598. Habibi Y., M. Mahrouz & M.R. Vignon, 2005. Isolation and structural characterization of protopectin from the skin of Opuntia ficusindica prickly pear fruits. Carbohydr Polymers 60, 205-213. Habibi Y., M. Mahrouz & M.R. Vignon, 2003. Isolation and structure characterization of a (4-O-methyl-D-glucurono)-D-xylan from the skin of Opuntia ficus-indica prickly pear fruits. J Carbohydr Chem 22, 33 1-337. Habibi Y., M. Mahrouz, M.-F. Marais & M.R. Vignon, 2004. An arabinogalactan from the skin of Opuntia ficus-indica prickly pear fruits. Carbohydr Res 339, 1201-1205. Hansen, R.K., Oesterreich, S., Lemieux, P., Sarge, D.K., Fuqua, S.A.W. 1997. Quercetin inhibits heat shock protein induction but not heat shock factor DNA-binding in human breast carcinoma celles. Biochem. Biophy Res. Comm., 239: 851-856. Hegwood D.A., 1990. Human health discoveries with Opuntia sp. (prickly pear) indica (L.) Mill. cladodes in the wound-healing process. J.HortSci 25, 13151316. Jonas, A., Rosenblat, G., Krapf, D., Bitterman, W., Neeman, I. 1998. Cactus flower extracts may prove beneficial in benign prostatic hyperplasia due to inhibition of 5 alpha

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reductase activity, aromatase activity and lipid peroxidation. Urol. Res. 26(4): 265-70 Kelloff, G.J., Sigman, C.C., Greenwald, P. 1999. Cancer chemoprevention: progress and promise. Eur. J. Cancer, 35: 2031-2038. Kuti J.O., 2004. Antioxidant compounds from four Opuntia cactus pear fruit varieties. Food Chem 85, 527-533. Kuti, J.O. 1992. Growth and compositional changes during the development of prickly pear fruit. J. Hort. Sci. 67: 861-868. Laurenz, J. C., Collier, C. C., Kuti, J. O., 2003. Hypoglycaemic effect of Opuntia lindheimeri Englem. in a diabetic pig model. Phytother. Res., 17, 2629. Lee B., J.E. Hyun, D.W. Li & Y.I. Moon, 2002. Effects of Opuntia ficus-indica var. Saboten stem on gastric damages in rats. Arch Pharm Res 25, 6770. Lee E.B., J.E. Hyun, D.W. Li & Y.I. Moon, 2001. The effect of Opuntia ficus-indica var. saboten fruit on gastric lesion and ulcer in rats. Nat Prod Sci 7, 90-93. Lee H., J.-S. Yoon, B.H. Lee, B.W. Choi & K.H. Park, 2000. Screening of the radical scavenging effects, tyrosinase inhibition, and anti-allergic activities using Opuntia ficus indica. Kor J Pharmacogn 31, 412415. Lee, E. B., Hyun, J. E., Li, D. W., Moon, Y. I., 2001. The effect of Opuntia ficus-indica var. saboten fruit on gastric lesion and ulcer in rats. Nat. Prod. Sci., 7, 9093. Lee, J.-C., Kim, H. R., Kim, J., Jang, Y.-S., 2002. Antioxidant property of an ethanol extract of the stem of Opuntia ficus indica var. saboten. J. Agric. Food Chem., 50, 6490 6496. Lee, Y. C., Hwang, K. H., Han, D. H., Kim, S. D., 1999. Compositions of Opuntia ficus-indica. Kor. J. Food Sci. Technol., 29, 847853. Meckes-Lozyoa, M., Roman-Ramos, R., 1986. Opuntia streptacantha, a coadjutor in the treatment of Diabetes mellitus. Am. J. Chinese Med., 14, 116118. Meyer, B. N., Mohamed, Y. A. H., McLaughlin, J. L., 1980. b-Phenethylamines from the cactus genus Opuntia. Phytochemistry,19, 719720 Moreno Alvarez M.J., C. Medina, L. Antn, D. Garca, D.R. Beln Camacho, 2003. Uso de pulpa de tuna (Opuntia boldinghii) en la elaboracin de bebidas ctricas pigmentadas. Interciencia 28, 539-543. Moreno P.R. & C.F. Flores, 1996. The world cactus pear market. J Profess Assoc Cactus Dev 1, 7586. Munoz de Chavez, M., Chavez, A., Valles, V., and Rodlan, J.A. 1995. The nopal : a plant of manifold qualities. World Rev. Nutr. Diet. 77, 109-134. Nerd A., M. Dumoutier & Y. Mizrahi, 1997. Properties and post-harvest behavior of the vegetable cactus Nopalea cochenillifera. Postharv Biol Technol 10, 135-143. Nieto, M., 1987. Alcaloides de Cactaceas. Estudio de cinco especies argentinas. Anales Asoc.

Quim. Argentina, 75, 11 13. Nobel, P. S., De la Barrera, E., 2003. Tolerances and acclimation to low and high temperatures for cladodes, fruits and roots of a widely cultivated cactus, Opuntia ficusindica. New Phytol.,.157,.271279. Nobel, P. S., Pimienta-Barrios, E., Zaeudo Hernandez, J., Ramirez-Hernandez, B., 2002. Historical aspects and net CO2 uptake for cultivated Crassulacean acid metabolism plants in Mexico. Ann. Appl. Biol., 140, 133 142. Park E.-H., J.H. Kahng & E.-A. Paek, 1998. Studies on the pharmacological actions of cactus: identification of its anti-inflammatory effect. Arch Pharm Res 21, 3034. Park E.H., J.H. Kahng S.H. Lee & K.H. Shin, 2001. An anti-inflammatory principle from cactus. Fitoterapia 72, 288-290. Qiu, Y., Chen, Y., Pei, Y., Matsuda, H., Yoshikawa, M., 2002. Constituents with radical scavenging effect from Opuntia dillenii, Structures of new a-pyrones and flavonolglycoside. Chem. Pharmaceut. Bull., 50, 15071510. Ramadan M.F. & J.-T. Mrsel, 2003a: Oil cactus pear (Opuntia ficus-indica L.). Food Chem 82, 339345. Ramadan M.F. & J.-T. Mrsel, 2003b: Recovered lipids from prickly pear [Opuntia ficus-indica (L.) Mill] peel: A good source of polyunsaturated fatty acids, natural antioxidant vitamins and sterols. Food Chem 83, 447456. Reynolds, T., Dweck, A. C., 1999. Aloe vera leaf gel: a review update. J. Ethnopharmacol., 68, 337. Senz C., E. Seplveda & B. Matsuhiro, 2004. Opuntia spp mucilage's: a functional component with industrial perspectives. J Arid Environ 57, 275-290. Senz C., 2002. Cactus pear fruit and cladodes: a source of functional components for foods. Acta Hort 581, 253-263. Senz-Hernandez C., J. Corrales-Garcia & G. Aquino-Perez, 2002. Nopalitos, mucilage, fiber, & cochineal. In: Cacti. Biology and Uses. Eds: Nobel PS, University of California Press, Berkeley, Los Angeles, 211-234. Senz, C. 2006. Utilizacion agroindustrial del nopal. Boletin de servicios agricolas de la FAO, 162: 164pp. Salt, T. A., Tocker, J. E., Adler, J. H., 1987. Dominance of D5-sterols in eight species of the Cactaceae. Phytochemistry, 26, 731 737. Shahidi F., P.K. Janitha & P.D. Wanasundara, 1992. Phenolic antioxidanta, Critical previous term Reviews next term. Food Sci Nutr 32, 67 103. Shapiro, K., Gong, W. C., 2002. Natural products used for diabetes. J. Am. Pharmaceut. Assoc., 42, 217226. Sloan, E. 1994. Top ten trends to watch and work

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on. Food Tech. 7, 89-100. Stintzing F.C. & R. Carle, 2005. Cactus stems (Opuntia spp.): A review on their chemistry, technology, and uses. Mol Nutr Food Res 49, 175-194. Stintzing F.C. & R. Carle, 2004. Functional properties of anthocyanins and betalains in plants, food, and in human nutrition. Trends Food Sci Technol 15, 1938. Stintzing F.C., A. Schieber & R. Carle, 2001. Phytochemical and nutritional significance of cactus pear. Eur Food Res Technol 212, 396-407. Stintzing F.C., K.M. Herbach, M.R. Mosshammer, R. Carle, W.G. Yi, S. Sellappan, C.C. Akoh, R. Bunch & P. Felker, 2005. Color, betalain pattern, and antioxidant properties of cactus pear (Opuntia spp.) clones. J Agric Food Chem 53, 442-451. Stintzing, F. C., Schieber, A., Carle, R., 2003. Evaluation of colour properties and chemical quality parameters of cactus juices. Eur. Food Res. Technol. 216, 303311. Tapiero H., K.D. Tew, G.N. Ba & G. Mathe, 2002. Polyphenols: Do they play a role in the prevention of human pathologies? Biomed Pharmacother 56, 200207. Tesoriere L., D. Butera, A.M. Pintaudi, M. Allegra & M.A. Livrea, 2004. Supplementation with cactus pear (Opuntia ficus-indica) fruit decreases oxidative stress in healthy humans: a comparative study with vitamin C. Am J Clin Nutri 80, 391-395. Tesoriere L., D. Butera, D. DArpa, F. Di Gaudio, M. Allegra, C. Gentile & M.A. Livrea, 2003. Increased resistance to oxidation of betalainenriched human low density lipoproteins. Free Radic Res, 37, 689696. Tesoriere L., D. Butera, M. Allegra, M. Fazzari & M.A. Livrea, 2005b. Distribution of betalain pigments in red blood cells after consumption

of cactus pear fruits and increased resistance of the cells to ex vivo induced oxidative hemolysis in humans. J Agric Food Chem 53, 1266-1270. Tesoriere L., M. Fazzari, M. Allegra & M.A. Livrea, 2005a. Biothiols, taurine, and lipid-soluble antioxidants in the edible pulp of Sicilian cactus pear (Opuntia ficus-indica) fruits and changes of bioactive juice components upon industrial processing. J Agric Food Chem 53, 785 1-7855. Veronesi U., G. De Palo, E. Marubini, A. Costa, F. Formelli, L. Mariani, A. Decensi, T. Camerini, M.R. Del Turco, M.G. Di Mauro, M.G. Muraca, M. Del Vecchio, C. Pinto, G. D'Aiuto, C. Boni, T. Campa, A. Magni, R. Miceli, M. Perloff, W.F. Malone & M.B. Sporn, 1999. Randomized trial of fenretinide to prevent second breast malignancy in women with early breast cancer. J Natl Cancer Inst 91, 18471856. Viegi L., A. Pieroni, P.M. Guarrera & R. Vangelisti, 2003. A review of plants used in folk veterinary medicine in Italy as basis for a databank. J Ethnopharmacol 89, 221-244. Wolfram, R. M., Budinsky, A., Efthimiou, Y., Stomatopoulos, J., Oguogho, A., Sinzinger, H., 2003. Daily prickly pear consumption improves platelet function. Prostagland. Leukotr. Ess. Fatty Acids, 69, 6166. Wolfram, R. M., Kritz, H., Efthimiou, Y., Stomatopoulos, J., Sinzinger, H., 2002. Effect of prickly pear (Opuntia robusta) on glucose- and lipid-metabolism in nondiabetics with hyperlipidemia a pilot study. Wiener Klin. Wochenschr., 114, 840846. Zou D.M., M. Brewer, F. Garcia, J.M. Feugang, J. Wang, R. Zang, H. Liu & C.P. Zou, 2005. Cactus Pear - a Natural Product in Cancer Chemoprevention. Nutr J, 4.

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Antioxidant Properties Of Cactus Products

Mnica A. Nazareno1 and Evangelina Gonzlez2
(1) Faculty of Agronomy and Agroindustries. (2) Faculty of Forestal Sciences. Nacional University of Santiago del Estero. Av. Belgrano (S) 1912. Santiago del Estero. Argentina. TE: 0385-4509528. E-mail: nazareno@unse.edu.ar

Abstract
Numerous investigations relate free radicals with the origin of serious diseases. A diet rich in fruits and vegetables is associated to lower incidences of cancer and heart illness. Such beneficial properties of natural foods have been assigned not only to their vitamin content but also to the presence of antioxidants, substances capable to scavenge free radicals. Cactus fruits have been reported as excellent sources of antioxidants. Several studies reveal that the antioxidant capacity is due to its vitamin C content as well as to the presence of polyphenols and water-soluble pigments named betalains. Fruits of cacti present moderate free radical scavenging ability, and values are similar to those of other season fruits. When cactus fruits were minimally processed or stored under refrigerated conditions, no important decreases in TEAC values were found neither significant vitamin losses were detected. Extracts obtained from cactus pear fruits where also used as additives to prevent oxidation of fish oil and its emulsions showing good thermal stability. In vivo assays have demonstrated that cactus fruit consumption improve the oxidative status of healthy individuals. Betalains present in cactus pear fruits have been reported to increase the resistance to human LDL oxidation. In addition, the regular intake of O. robusta has been reported to reduce oxidative injury in patients with hypercholesterolemia. For these reasons, cactus fruits are excellent sources of bioactive substances and so, it is highly recommendable to incorporate them to a normal diet as fresh fruits. Besides, they are potentially utilizable in the elaboration of high added value products such as cosmetics, nutraceutics or dietary supplements.

increasing tremendously these days. These substances present the ability to scavenge free radicals or other non radical oxidant species delaying or retarding the oxidative damage produce by them. Numerous investigations indicate that free radicals are involved in oxidation processes resulting in cell aging and the origin of serious pathologies such as cancer, heart diseases, degenerative illness of the nervous system and malaria (Halliwell and Gutteridge, 1992; Trush and Kensler, 1991). Cigarette smoke, pollutants, organic solvents, anesthetic and pesticides are some important sources of free radicals (Halliwell and Gutteridge, 1989). They are also formed by exposure to sunlight, X-rays and ozone. Actually, free radicals are no intrinsically harmful. In fact, they are produced in the organism in moderate amounts as a mechanism of defense against bacteria and virus. Once they complete their duty, they are easily neutralized by the organism itself. Optimum health state depends mainly on such balance. When the production of oxidant species exceeds the body's natural antioxidant action, oxidative stress occurs causing damage to cellular biomolecules like nucleic acid, enzymes, lipids, carbohydrates and, consequently, may affect immune functions resulting in a variety of diseases (Ames et al., 1993). Oxidative stress is one of the major factors responsible for DNA damages resulting in mutagenic changes due to harmful effects of reactive oxygen species (Zhao et al., 2005). Diverse defense mechanisms take place in the body towards the attack of such deleterious species that involve enzymes, vitamins and other natural substances that avoid or retard oxidative damage of cell constituents (Yu, 1994).

Introduction
The interest on natural antioxidants is

18

OXIDATIVE STRESS

Figure 1. Oxidative stress

Several reports indicate that a diet rich in fruits and vegetables are related to lower incidences of heart conditions and some types of cancer suggesting that this kind of diet has positive effects on health (Bazzano et al., 2002). These beneficial effects have been associated to the action of some antioxidant components present in that natural food and not only to their vitamin content (Terry et al., 2002, Hertog et al., 1995). Nowadays, discovery of new compounds obtained from natural sources with high antioxidant activity is a constant challenge for researchers. There are many groups of compounds such as carotenoids, betalains, flavonoids and other phenolic compounds occurring in nature with these properties. Many of them are derived from plants and widely spread in food so they are incorporated frequently as part of a diet. In the search for health promoting substances, the assessment of a relative activity scale is necessary. However, antioxidants present different behaviour depending on the nature of the oxidative species since different mechanisms of action take place for their deactivation or scavenging. A large series of methods has been proposed to evaluate in vitro the intrinsic activity of the antioxidants towards different reactive species formed in chemical and biological model

systems. Besides, health protective and disease-preventive effects of natural substances, in pure form or as components of food, have also been measured in vivo.

In vitro measurements
Numerous methodologies have been developed to determine the antioxidant capacity of synthetic and natural pure compounds (Schlesier et al., 2002) extracts made from fruits, herbs and vegetables as well as drinks like wine, tea and juices (Pellegrini et al., 2000). Some of them determine specifically the free radical scavenging capacity or antiradical ability of the antioxidants by measuring radical consumption by effect of the addition of them. One of the most frequently chosen techniques by its simplicity is the evaluation of synthetic radical disappearance as the 1,1-diphenyl-2-pycrilhydrazyl radical (DPPH) or the 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation or ABTS+.. It measures the ability of pure substances or crude extracts for trapping free radicals by donating hydrogen atoms or electrons producing, as a consequence, the bleaching of the coloured radical solution and the formation of less reactive species from the antioxidant (Brand-Williams et al., 1995). The reaction is monitored by UV-Visible Spectrophotometry measuring the decrease in

19

absorbance concentration.

associated

to

radical

Other methodologies determine the capacity of natural extracts to retard the oxidation of a chemical system by monitoring the consumption of oxidizable species or the formation of its oxidation products. A frequently used assay for antioxidant capacity determination of fruits, vegetables and other foods is the evaluation of the ability of their extracts to trap peroxyl radical and therefore, inhibit unsaturated lipid oxidation named lipid peroxidation (Cao et al., 1993; Chaillou and Nazareno, 2006). All of these techniques are in vitro determinations and they represent indirect measurements of the real action that a substance can exert in the body to prevent diseases causes by free radicals. This is a very controversial subject, however, scientific community agrees, in general, that there is a relation between the consumption of antioxidants, especially those with high antiradical activity and the beneficial effect in health previously mentioned. As it was shown, numerous different techniques are proposed to measure the antiradical ability of a substance, thus, an impressive diversity of forms of expression of this activity also exists in literature (Prez et al., 2000). This variety makes sometimes difficult the comparison between results expressed with different reference compound or obtained from diverse experimental designs. One of the several forms of expression of the results obtained from free radical scavenging techniques is the percentage of Antiradical Activity (AAR %) that considers the fraction of radical consumed by a given amount of sample. In the case of fruits and their juices where vitamin C is one of the major active substances, other option to display the results easily comparable and meaningful, is the Vitamin C Equivalent Antioxidant Capacity (VCEAC) which represents the amount of ascorbic acid that present the same capacity of the total extract (Kim et al., 2002). Other very commonly used expression is Trolox Equivalent Antioxidant Capacity (TEAC) which compares the ability of a substance or extract to donate a hydrogen atom or an electron compared with Trolox, a hydro-soluble synthetic derivative of tocopherol. TEAC assay is based on the scavenging ability of antioxidants to the long-life radical anion ABTS.+. A common parameter to display the antioxidant activity is the Oxygen Radical Absorbance Capacity (ORAC) where inhibition

grade as well as oxidation reaction lag time are combined in the same value. ORAC measures antioxidant inhibition of peroxyl radical induced oxidations and, thus, it shows a classical radical chain breaking antioxidant activity by H atom transfer. In the basic assay, a peroxyl radical reacts with a fluorescent probe to form a non fluorescent product, which can be quantified easily by monitoring the residual fluorescence of the system. ORAC values are usually reported as Trolox equivalents.

Cactus Fruits as antioxidant sources

Many cactus species produce edible fruits, often consumed as fresh fruits as well as dried food or in processed products as marmalades, jams, ice-creams, juices or syrup. Cactus fruits have been reported as excellent sources of bioactive substances such as vitamins and antioxidants. One of the more valuable contributions of cactus fresh fruits to a diet is their vitamin C content. Besides, they have other powerful antioxidants such as polyphenols and betalains. The last ones are also responsible for the beautiful characteristic colors of the fruits. Betalains are natural hydrosoluble pigments. Considering their chemical structures, betalains are present in two different groups, betaxanthins, yellow and orange colored substances, and betacyanins, red to purple colored substances (Galati et al., 2003). Most known examples of these two families in cactus pear fruits are indicaxanthin for the former and betanin for the latter. Betanin is famous for being also the typical colour of red beet. Concerning to flavonoids, many types of them have been reported in Opuntia sp. and an important role has been proposed for their presence as the main responsible of the antioxidant power of its extracts (Lee and Lim, 2000). Their chemical structure and concentration are very variable and depend on the variety, ripening stages and the kind of tissue of the plants (Wallace, 1986) 3-Oglycosilated flavonols, dihydroflavonols, flavonones and flavanols have been found in Cactaceae plants and fruits (Kuti, 2000). Galati et al., 2003, analized Opuntia ficus indica (L.) Mill. juices (95% yellow and 5% red cultivars) and they identified the presence of major flavonoids, isorhamnetin triglycoside, rutin and kaempferol, as the main watersoluble constituents. Total flavonoid content corresponds to 652.5 g/mL and ascorbic acid concentration is 26.9 mg/ 100 mL of juice.

20

Antiradical activities of fruit juice and the fruit hydro-soluble fraction were measured, and the aliquot that produce a 50 % decreased of DPPH absorbance (IC50) were 6.75 L and 7.68 L, respectively. Fruit organic fraction has not shown any antiradical activity. The activity exhibit by juice and hydro-soluble extract are assigned to polyphenols as ferulic acid and flavonols, ascorbic acid and betalains. Tesoriere et al., 2004, reported that 100 g of cactus pear fruit flesh from yellow Sicilian cultivars contain 29 mg of vitamin C, 1.21 mg of betanin, 9.3 mg of indicaxanthin, 80 g of tocopherol and 1.5 g of -carotene. Major constituent of the antioxidant fraction is vitamin C followed by betalains. Same results where reported by Butera et al., 2002, indicating that Opuntia ficus-indica cultivars red, yellow and white varieties present ascorbic acid as main component (30 mg/ 100 g of edible pulp) followed by betalains, mainly indicaxanthin and betanin. Yellow and white varieties are rich in indicaxanthin, 8.42 and 5.86 mg/ 100 g of fresh pulp, respectively, and red one in betanin, 5.12 mg/ 100 g of fresh pulp, corresponding in this case to a 66 % of the total betalain content. Flavonoid content is about 0.237 g of flavonols for 100 g of fresh pulp. Total antioxidant activities towards ABTS+. radical of aqueous extracts are 5.31 for yellow variety, 4.20 for the red one and 4.36 for the white one expressed as mol of Trolox equivalent per g of edible pulp, that is to say the most powerful one trapping ABTS+. free radicals was the yellow one. More recently, antioxidant activity of fruit extracts for 4 varieties of Opuntia spp (O. ficusindica, O. lindheimeri, O. streptancantha and O. stricta var. stricta, green, purple, red and yellow peels, respectively) were reported. These properties were attributed to the presence of flavonoids, mainly to quercetin derivatives, kaempferol and isorhamnetin, ascorbic acid and carotenoids (Kuti, 2004). Flavonoid content ranges between 9.8 to 93.5 g per g of fresh fruit and the highest level was detected for purple peel fruits. Ascorbic acid content was very variable, 10 to 111 g/ g for purple fruits and 23 to 792 g/ g for red ones. Carotenoid contents present highest values for yellow ones (6.0 to17.7 g/g) and the lowest for the green ones (1.2 to1.7 g/g). Antioxidant

activity measured as ORAC values correlated with flavonoid contents and not with ascorbic acid or carotenoid concentrations. Stintzing et al. (2005), evaluated antioxidant activity as ORAC and TEAC values for O. ficus-indica and O. robusta clones and assigned it to polyphenols, betalains and ascorbic acid present in these fruits. They reported a strong correlation between this activity with polyphenol content and in lower degree with betalains. Betaxanthin and betacyanin contents measured for 100 g of fresh fruits were 6.59 and 114.04 mg, respectively. These values coincide with previous literature data (Odoux and Dominguez-Lopez, 1996; Castellar et al., 2003; Stintzing et al., 2003, Stintzing et al., 2005). Ascorbic acid contents determined in this article for Opuntia fruits and their juices were 10 folds lower than other reports (Kuti, 2004; Butera et al., 2002; Gurrieri et al., 2000) and so properties as TEAC and ORAC values compared with other fruits and vegetables. TEAC values of cactus juices are close to those of red wine and green tea but they are lower than those of blueberries or pomegranate juices. Main active substances for cactus pears are polyphenols, followed by betalains and finally ascorbic acid (Stintzing et al., 2005). Free-radical scavenging activities for different Opuntia sp. fruits cultivated in semiarid region of Argentina were determined in order to establish a relative comparative scale towards the same radical. Fruits evaluated were O. sulphurea, Cereus forbesii, Trichocereus pasacana, O. robusta, Stetsonia coryne, O. ficus indica (pinkish, dark purple, purple and green), O. crassa, H. pomanensis, O. anacantha (retrorsa and utkilio), O. megacantha (white and yellowish) and Opuntia spp. (purple). Highest antioxidant activity was obtained for O. sulphurea followed by C. forbesii. The former presented a noticeable high value being surpassed only by some fruits well recognized as rich in antioxidant such as strawberries, kiwis and blackberries. Figure 2 shows that different cactus fresh fruits present moderate antioxidant ability, and values are similar to those of citric and other season fruits.

21

ANTIRADICAL ACTIVITIES OF CACTUS FRUITS

1.5

Vitamin C equivalent (mg/g)

1.3 1.1 0.9 0.7 0.5 0.3 0.1 -0.1

0.25 1.02 0.85 0.76 0.72 0.68 0.57 0.57 0.53 0.52 0.51 0.5 0.50 0.48 0.470.46 0.54 0.46 0.27 0.46

Tradicional fruits
0.72 0.54

O. megacantha (yellowish)

O. anacantha var. Ret.

O. anacantha var.utk.

S. coryne

O.f-i dark (purple)

H. pomanensis

O.f-i (pinkish)

C. forbesii

O.spp.

grapefruit

O.crassa

apricot

tangerine

O. sulphurea

T. pasacana

O.robusta

O.f-i (purple)

Figure 2. Antiradical activities of cactus fruit Bioactive substance contents were also measured for different cactus fruits from Argentina (Coria Cayupn et al., 2007). Analysis per 1 g of Opuntia spp, O. megacantha and O. ficus-indica purple and dark purple fresh fruit have showed ranges for vitamin C content of 0.26-0.48 mg, and for total polyphenol content of 0.54-1.12 mg. Considering that the Dietary Reference Intake for vitamin C is about 60 mg per day, the portion of cactus fruit containing this amount of vitamin ranges 125 to 230 g. Vitamin C values are similar to those reported for different species and varieties of Opuntia corresponding to 0.121-0.815 mg/g of fresh fruits (Kuti, 2004; Butera et al., 2002). Besides, O. spp. purple, O. ficus-indica dark purple and O. megacantha yellowish present the highest total polyphenols contents among the samples studied whereas O. megacantha white has the lowest one. From these results, the antioxidant activity correlates with polyphenol contents more than with vitamin C content (Cai et al., 2003), O. spp. purple and O. ficus-indica dark purple present the highest concentration of betalains followed by O. ficus-indica purple. Besides, betacyanin concentration is much higher than that of betaxanthins. A different situation occurs with O. megacantha yellowish pigment composition, where betaxanthin concentration is three folds of that of betacyanins. -carotene presence was also detected. Carotenoid content is 31 g/ g of fresh fruit, corresponding to a Vitamin A value of 5.2 RE/g. The fruits studied present good inputs of vitamin C to a diet and this substance is one of the responsible to the total antioxidant activity. O. ficus-indica purple has the highest concentration of vitamin C. Cactus fruits are also rich in polyphenolic compounds and the free radical scavenging ability of fruit extract correlates as well with their concentration.

Uses of cactus pear extract as antioxidant additives

Extracts obtained from cactus pear fruits where used experimentally as additives to prevent oil oxidation. Opuntia ficus-indica fruit extracts were added to a lipid system formed by fish oil or its emulsions and the lipid peroxidation was monitored by conjugated dienes formation.
22

O. megacantha (white)

O.f-i (green)

orange

peach

pear

Oxidation reaction rate decreased by extract addition and inhibitory effect of 0.1% extract after storage during 12 days was comparable with that of BHA and slightly higher than that of BHT. Extract addition produce a significant reduction in peroxide index compared with control system. O. ficus indica fruit extracts were heated at 50, 75 and 100 C for 15 min without antioxidant activity losses. Such heat stability makes them good alternatives compared to less thermostable compounds as -tocopherol (Siriwardhana and Jeon, 2004).

Processing effect in antioxidant activity of cactus fruits

Piga et al. (2003) studied minimally processed cactus pear fruits. Peeled fruits were kept at 4C and total phenol contents, vitamin C and antioxidant activity were monitored for 3, 6 and 9 days. Results showed that no significant losses of ascorbic acid took place in 9 days. This was attributed by the protective effect of polyphenols which concentration decreased in 6 days. No important decreases in TEAC values were found, and a strong correlation (0.978) between this antioxidant activity and ascorbic acid content was observed confirming ascorbic acid as one of the most important antioxidant in fruits. Studies directed to evaluate possible changes

in the content of bioactive substances and nutritional values of cactus pear fruits conserved under refrigerated storage were carried out by Coria Cayupn et al. (2007). When Opuntia ficus-indica "Yellow without spines" is exported from Argentina to Northern hemisphere countries, the transfer of the fruits is carried out by sea (5-8C) and delays approximately 3-4 weeks. In order to evaluate the real amount of health-promoting substances present in the fruits when they arrive to external markets and consumers, post-harvest and transport conditions were simulated during the same period of time. Possible changes in antiradical activity, vitamin C and total polyphenol content were evaluated at the harvest date and after their storage at 8C for 3 and 4 weeks. This antiradical ability of cactus pear fruits remains after 4 weeks at 8C without significant changes. In general, an increase in the concentration of active substances is observed corresponding to the loss of water content of the fruits of almost 10 % in 4 weeks. Pulp humidity showed much smaller variations of approximately 4 %, in the mentioned period of time. Total polyphenol content at harvest time was 0.60 mg/g while reached 0.71 and 0.77 mg/g in 3 and 4 weeks respectively. No important vitamin losses were detected as shown in Figure 3. As conclusion, fruit keep their beneficial properties even after 4 weeks of refrigerated storage.

0,6 0,5 0,4 Vit C content 0,3 mg/g 0,2 0,1 0 0 3 Weeks 4

Vitamin C

VCEAC

Figure 3. Evolution of Vitamin C content and Antioxidant Activity for refrigerated storage of cactus pear fruits at 8C Many cactus species produce edible fruits, often consumed as fresh fruits as well as dried food or in processed products as marmalades, jams, ice-creams, juices or syrup. Antiradical activity measurements of these fruit derivatives show that, even after thermal treatments, marmalades and jams elaborated from cactus fruits retain this capacity. Opuntia ficus-indica syrup has an antioxidant activity equivalent to 102 mg of ascorbic acid per 100 g of product. Cereus forbesii Jam activity value was 74 mg/ 100 g, and O. robusta and O. anacantha marmalades, activity values were equivalent to 64 and 56 mg of vitamin C per 100 g. These are similar to those obtained for marmalades of traditional fruits as pears, oranges and

23

strawberries of 65, 35, and 73 mg of vitamin C per 100 g respectively. Retention of antiradical activity of cactus products is higher compared to those of other traditional fruit products. These processed products derived from cactus

fruits can be considered as functional food because of their good activity; they are available all the year long and well-accepted by most consumers.

Figure 4. Antioxidant activity of fruits products

In vivo assays
Although in vitro determinations of the intrinsic antioxidant activity of food are relevant, is important to consider that the factors that influence its efficiency in vivo are complex, thus, the results obtained in vitro cannot easily be extrapolated to in vivo systems. The effectiveness of an antioxidant depends on diverse factors as its bioavailability, the action site, the type of reactive species with which it will react and the possible synergistic effects with other compounds present in the food matrix. Among factors previously mentioned, one of most important is the bioavailability which mainly depends on the antioxidant releasing from the food matrix, its

stability in the gut flora, the chemical modifications that it may suffer in the intestine, its absorption through the intestinal wall towards the blood stream, the stability in the liver and its accessibility to the tissue to the target site (William et al., 2004). As well as for in vitro determinations, there are several methodologies developed to determine the in vivo activity of an extract or a substance. They basically imply the measurements of certain "markers" associated to the oxidative damage in body fluids, before and after changes in the diet or supplementation with an antioxidant extract (Aruoma, 2003). Some examples of these target molecules used to evaluate body oxidative level are shown in Table 1.

24

Table 1: Biomarkers of oxidative stress in humans Markers Description Isoprostanes have been described as F-prostaglandin isomers produced by araquidonic acid oxidation. Isoprostanes levels in vivo increased under oxidative stress conditions. Measurement of different groups of isoprostanes might be an approach to asses the relative rates of peroxidation of different PUFAs in vivo and emerged as one of the most reliable biomarkers. Determinations are carried out by CG-MS. MDA is the final degradation product of peroxides formed in the oxidation of free fatty acid or membranes. It can be detected in plasma as a biomarker for the in vivo lipidic oxidation. Its reaction with thiobarbituric acid is followed by the colour development of the formed adduct. Glutation is the most powerful intracellular antioxidant and its relation between its oxidized and reduced forms (GSH:GSSG) represents the antioxidant capacity of the cell. LDL oxidation is a free radical process leading to numerous structural and functional changes, crucial in atherosclerotic lesion formation. This reaction happens mainly between the walls of the blood vessels therefore, the LDL oxidative status reflects the oxidation state of the blood vessels. Fatty acid peroxidation conducing to conjugated diene formation can be detected by Spectrophotometry. Ref. (Roberts and Morrow, 2000; Tesoriere et al., 2004; Budinsky et al., 2001) (Cadenas, 2001; Halliwell and Grootveld, 1987; Janero, 1990) (Martnez Sarrasague et al., 2006).

F2-isoprostanes (8-epi-PGF2) in plasma or urine

Malondialdehyde (MDA) in plasma or serum Glutation redox status in red bood cells Low Density Lipoprotein (LDL) analysis in plasma

(Cadenas, 2001; Tesoriere et al., 2004).

Other interesting parameter for evaluation is the global bodys antioxidant status level after a special diet. As an example, after supplementation with cactus pear fruits to human, bioactive substance levels in plasma were determined measured as well as the antioxidant ability of this fluid was measured and expressed as TEAC values.

Evaluation of the Antioxidant activity in vivo of cactus products

Betalains, those pigments widely spread in cactus pear fruits have been reported to exert an important role in preventing oxidative stress injuries. Main interests of researches in this subject are focused to determination of betalain contents, their absorption and antioxidant effects in vivo. Since antioxidant efficiency in vivo fundamentally depends on its bioavailability, assays that tend to study those properties of betalains were carried out. Experiments have demonstrated that red beet betalains have been detected in urine after 2 or 3 h of ingestion, recovering 0.5 to 0.9 % of the total intake. Other interesting fact is that betalain detected in urine is recovered as its isomeric form isobetanin demonstrating that

the original molecule undergoes an isomerization process probably due to body high temperature (Kanner et al., 2001). Betalains have also been detected in blood after 3 h of ingestion of 500 g of fresh cactus pulps. After this period, pigment concentration was decreasing along time and almost disappeared in 8 h (Tesoriere et al., 2005). Betalain levels were also measured in human body fluids showed after intake of cactus pear fruits. Urine recovery after 12 hours was about 76 % for indicaxanthin and 3.7 % for betanin. Pigment peak concentrations were reached in plasma in 3 hours after ingestion for both of them and completely disappeared in12 h after ingestion (Tesoriere et al., 2004). Betalains have been reported to increase the resistance to human LDL oxidation (Tesoriere et al., 2003; Zakharova and Petrova, 1998). They are able to bind to low density lipoproteins (LDL) and this union prevent their oxidation. After fruit consumption, LDL extracted from plasma in this conditions indicated that 3 h after intake, indicaxanthin was incorporated in 98 pmol/mg of protein diminishing in 5 h and it complete disappeared in 8 h. Betanin was detected only 3 h after fruit consumption and lag time of LDL oxidation

25

induced by copper was longer compared with control. Ex vivo study evaluated the oxidative hemolysis of human red blood cells induced by free radicals. Blood samples were taken from volunteers after ingestion of cactus pear fruits. Results indicated that cell resistance to hemolysis was higher after 3 h of fruit intake corresponding to highest plasma level of betalains (Tesoriere et al., 2005). Besides, methanolic extract containing 1 to 5 mg of Opuntia ficus-indica pulp inhibited MDA formation in 4 h incubation of blood cells exposed to organic hydroperoxides as a free radical source (Butera et al., 2002). A comparative study was carried out to evaluate the effect of O. ficus indica intake and vitamin C supplementation in oxidative status of healthy volunteers (Tesoriere et al., 2004). Results showed that the consumption of 250 g of fresh fruit pulp as well as its equivalent, 75 mg vitamin C supplementation produce an increase of vitamin C and E levels. No changes were found in vitamin A content or TEAC value in plasma. Differences between both treatments become relevant in oxidative stress biomarker levels. After fruit intake, 8-epiPGF2 and MDA plasma values decreased in 30 and 75 %, respectively, while glutation redox-status relation (GSH:GSSG) in red blood cells was enhanced since the reduced form increased. LDL-conjugated dienes hydroperoxide level was significantly reduced and so LDL oxidation. Vitamin C supplementation has not produced changes in oxidative stress markers indicating that cactus fruit efficiency is due to AO combination including vitamin C and other co-nutrients as betalains. Budinsky et al. (2001), reported that regular ingestion of broiled pulp of O. robusta is able to reduce significantly in vivo oxidative damage in hypercholesterolemic patients. Injury was evaluated measuring 8-epi-PGF2 in plasma, serum and urine. Results showed that fruit ingestion had not influence in body weight neither in body fat content however total cholesterol level decreased specially in men. Values of 8-epi-PGF2 in plasma as well in serum and urine presented an important decrease being more noticeable in women. Compounds responsible for such bioactivity have not been yet established.

vitro systems as well as in vivo experiments. Cactus fruits have been reported as excellent sources of bioactive substances such as vitamins and antioxidants. Processed products elaborated from fruits are able to retain the antiradical capacity so they can be considered as functional foods available all year long. This ability to inhibit oxidation reactions are attributed to the presence of polyphenols, betalains and vitamin C as the main active compounds. Experimental in vivo results show that regular supplementation with cactus fruits in a normal diet enhances health status reducing oxidative stress effect.

Acknowledgements Authors are especially grateful to Prof. Judith Ochoa and her team work from INDEAS for fruit and plant material supply, technical support and valuable discussions. This work was supported by CONICET, ANPCYT and CICYT-UNSE. References
Ames, B. M.; Shigena, M. K.; Hagen, T. M., 1993, Oxidants, antioxidants and the degenerative diseases of aging. Proc. Natl. Acad. Sci. USA 90, 7915-7922. Aruoma, O. I., 2003, Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. Mutation Research, 9, 523524. Bazzano, L.A.; He, J.; Ogden, L.G.; Loria, C.M.; Vupputuri; S., Myers, L.; Whelton, P.K. 2002, Fruit and vegetable intake and risk of cardiovascular disease in US adults: the first national health and nutrition examination survey epidemiologic follow-up study. Am. J. Clin. Nutr. 76, 93-99. Brand-Williams, W.; Cuvelier, M.E.; Berset, C., 1995, Use of a free radical method to evaluate antioxidant activity. Lebensm.-Wiss. U.-Technol. 28, 25-30. Budinsky, A.; Wolfram, R.; Oguogho, A.; Elthimiou, Y.; Stamatopoulos, Y.; Sinzinger, H., 2001, Regular ingestion of Opuntia robusta lowers oxidation injury. Protaglandins, leucotrienes and essential fatty acids. 55, 45-50. Butera, D.; Tesoriere, L.; Di Gaudio, F.; Bongiorno, A.; Allegra, M.; Pintaudi, A. M.; Kohen, R.; Livrea, M. A., 2002, Antioxidant activities of Sicilian prickly pear (Opuntia ficus-indica) fruit extracts and reducing properties of its betalains: betanin and indicaxanthin J. Agric. Food Chem. 50, 6895-6901. Cadenas, E.; Packer, L. 2001, Handbook of Antioxidants. Marcel Dekker Inc. ISBN 0824705475. Cai, Y.; Sun, M.; Corke, H., 2003, Antioxidant

Conclusions
Protective effects towards oxidation processes exerted by fruits, juices and their processed products have been widely demonstrated in

26

activity of betalains from plants of the Amaranthaceae. J. Agric. Food Chem, 51, 2288-2294. Cao, G.; Alessio, H.; Cutler, H., 1993, Oxygenradical absorbance capacity assay for antioxidants. Free Rad. Biol. Med., 14, 303311. Castellar, R.; Obon, J.J.; Alacid, M.; FernndezLpez, J. A. 2003, Color properties and stability of betacyanins from Opuntia fruits J. Agric. Food Chem. 51, 2772-2776. Chaillou, L.L.; Nazareno, M.A., 2006, New Method to Determine Antioxidant Activity of Polyphenols. J. Agric. Food Chem. 54, 83978402. Coria Cayupn, Y.; Ochoa, M.J.; Nazareno, M.A., 2007, Health-Promoting Substance Contents and Antioxidant Properties of Opuntia sp. Fruits from Santiago del Estero, Argentina. Proceedings of the VI International Congress on Cactus pear and Cochineal, Brazil. Coria Cayupn, Y.; Targa, G.; Ochoa, M.J.; Nazareno, M.A., 2007, Bioactive substance content and antioxidant activity changes during cooled storage of yellow spineless cactus pears. Proceedings of the VI International Congress on Cactus pear and Cochineal, Brazil. Galati, E. M.; Mondello, M. R., Giuffrida, D.; Dugo, G.; Miceli, N.; Pergolizzi, S.; Taviano, M. F., 2003, Chemical Characterization and Biological Effects of Sicilian Opuntia ficusindica (L.) Mill. Fruit juice: antioxidant and antiulcerogenic Activity J. Agric. Food Chem. 51, 4903-4908. Gurrieri, S.; Miceli, L.; Lanza, C. M.; Tomaselli, F.; Bonomo, R. P.; Rizzarelli, E., 2000, Chemical caracterization of Sicilian prickly pear (Opuntia ficus-indica) and perspectives for the storage of its juice. J. Agric. Food Chem. 48, 5424-5431. Halliwell, B.; Grootveld, M., 1987, The measurement of free radical reactions in humans. FEBS Lett. 9, 213-215. Halliwell, B.; Gutteridge, J.M.C., 1989, Free nd Radicals in Biology and Medicine. 2 Edition. Oxford University Press, London. Halliwell, B.; Gutteridge, J.M.C.; Cross, C.E., 1992, Free Radicals, Antioxidants and human diseases, Where are we now? J. Lab. Clin. Med. 119, 598-620. Hertog, G. L.; Kromhout, D.; Aravanis C.; Blackburn, H.; Buzina, R.; Fidanza F., 1995, Flavonoid intake and long term risk of coronary heart disease and cancer in the seven country study. Arch. Intern. Med. 155, 381-386. Janero, D.R., 1990, Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Rad. Biol. Med. 9, 51540. Kanner, J.; Harel, S.; Granit, R., 2001, Betalains- A new class of dietary cationized antioxidants. J. Agric. Food Chem., 49, 5178-5185.

Kim, D.O.; Lee, K.W.; Lee, H. J.; Lee, C. Y., 2002, Vitamin C Equivalent Antioxidant Capacity of Phenolic Phytochemicals. J. Agric. Food Chem. 50, 3713-3717. Kuti, J. O., 2000, Antioxidant activity of Opuntia cactus pear. Hort Science 35, 433. Kuti, J. O., 2004, Antioxidant compounds from four Opuntia cactus pear fruit varieties. Food Chem. 85, 527-533. Lee, J.C.; Lim, K. T., 2000, Effects of cactus and ginger extracts as dietary antioxidants on reactive oxidant and plasma lipid level. Food Sci. Biotechnol. 9, 83-88. Martnez Sarrasague, M.; Barrado, D. A.; Zubillaga, M.; Hager, A.; De Paoli, T. Boccio, J., 2006, Conceptos actuales del metabolismo del glutatin. Utilizacin de los istopos estables para la evaluacin de su homeostasis. Acta Bioqum. Cln. Latinoam., 40, 45-54. Odoux, E.; Dominguez-Lpez, A., 1996, Le figuier de Barbarie: une source industrielle de btalanes ? Fruits 51, 61-78. Pellegrini, N.; Simonetti, P.; Gradana, C.; Brenna, O.; Bririghenti, F.; Pietta, P., 2000, Polyphenol content and Total Antioxidant Activity of Vini Novelli (Young Red Wines) J. Agric. Food Chem. 48, 732-735. Prez, D. D.; Leigthon, F.; Aspee, A.; Aliaga, C.; Lissi, E. A., 2000, A comparison of methods employed to evaluate antioxidant capabilities. Biol. Res. 33, 71-77. Piga, A.; Del Caro, I.; Pinna, M.; Agabbio 2003, Changes in ascorbic acid, polyphenol content and antioxidant activity in minimally processed cactus pear fruits Lebensm.-Wiss. U.-Technol. 36, 257262 Roberts, L.J.; Morrow, J.D., 2000, Measurement of F2-isoprostanes as an index of oxidative stress in vivo. Free Rad. Biol. Med. 28, 505 513. Schlesier, K.; Harwat, M.; Bhm, V.; Bitsch, R., 2002, Assessment of antioxidant activity using different in vitro methods Free Radical Res. 36, 177-187. Siriwardhana; N.; Jeon, Y., 2004, Antioxidative effect of cactus pear fruit (Opuntia ficusindica) extract on lipid peroxidation inhibition in oils and emulsion model systems. Eur Food Res Technol. 219, 369376 Stintzing, F.C.; Herbach, K. M.; Mosshammer, M. R.; Carle, R. Yi, W.; Sellappan, S.; Akoh, C.C.; Bunch, R.; Felker, P., 2005, Color, Betalain Pattern, and Antioxidant properties of Cactus Pear (Opuntia spp.) Clones J. Agric. Food Chem. 53, 442-451. Stintzing, F.C.; Schieber, A.; Carle, R., 2003, Evaluation of colour properties and chemical quality parameters of cactus juices Eur. Food Res. Technol. 216, 303-311. Stintzing, F.C.; Schieber, A.; Carle, R., 2001, Phytochemical and nutritional significance of cactus pear Eur. Food Res. Technol. 212, 396-407

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Terry, P.; Lain, M.; Miller, A.B.; Howe, G.R., Rohan, T.E., 2002, Dietary carotenoids and risk of breast cancer. Am. J. Clin. Nutr. 76, 883888. Tesoriere, L.; Butera, D.; Pintaudi, A. M.; Allegra, M.; Livrea, M. A., 2004, Supplementation with cactus pear (Opuntia ficus-indica) fruit decreases oxidative stress in healthy humans: a comparative study with vitamin C13. Am. J. Clin. Nutr. 80, 391395. Tesoriere, L.; Allegra, M.; Butera, D.; Livrea, M. A., 2004, Absorption, excretion and distribution of dietary antioxidant betalains in LDLs: potential health effects of betalains in humans. Am. J. Clin. Nutr., 80, 941-945. Tesoriere, L.; Butera, D.; Allegra, M.; Fazzari, M.; Livrea, M. A., 2005, Distribution of betalain pigments in red blood cells after consumption of cactus pear fruits and increased resistance of the cells to ex vivo induced oxidative hemolysis in humans. J. Agric. Food Chem. 53, 1266-1270. Tesoriere, L.; Butera, D; DArpa, F.; Di Gaudio, M.; Allegra, M.; Gentile, C.; Livrea, M. A. 2003, Increased resistance to oxidation of betalain-

enriched human low density lipoproteins. Free Radic. Res. 37, 689-696. Trush, M. A.; Kensler, T. W., 1991, Role of free radicals in carcinogen activation. In Oxidative stress. Oxidants and antioxidants 277-318; Helmut Sies, Ed.; Academic Press; San Diego, CA. Wallace, R. S., 1986, Biochemical taxonomy and the Cactaceae. Cactus & Succulent J. (USA) 58, 35-38. William, R.J.; Spencer, J.P.E; Rice-Evans, C., 2004, Flavonoids: antioxidants or signaling molecules? Free Rad. Biol. Med. 36, 838849. Yu, B. P., 1994, Cellular defenses against damage from reactive oxygen species. Biol. Rev. 74, 139-162. Zakharova, N. S.; Petrova, T. A., 1998, Relationships between the structure and antioxidant activity of certain betalains. Appl. Biochem. Microbiol. 34, 182-185. Zhao, C.; Dodin G.; Yuan C.; Chen H.; Zheng R.; Fan, Z. J. B., 2005, In vitro protection of DNA from Fenton reaction by plant polyphenols verbascoside. Biochim. Biophys. Acta, 1723, 114-123.

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Antihyperlipidemic Effect Of Cactus Products

Helmut Sinzinger
1 2

The Austrian-Greek Atherosclerosis Prevention Initiative (AGAPI),

Wilhelm Auerswald-Atherosclerosis Research Group (ASF) Vienna, Austria

Nadlergasse 1, A-1090 Vienna, Austria. Tel +43 1 4082633; Fax: +43 1 4081366; E-mail: helmut.sinzinger@chello.at

Abstract
Used in traditional medicine since long, Opuntia and compounds derived from it have shown under experimental conditions as well as in human that a variety of metabolic parameters are beneficially affected. The underlying responsible compound, however, fruit or cactus itself, which type, the preparation, the amount necessary per day are not well defined. There is, however, certain evidence that Opuntia consumption besides improving glucose metabolism exerts multiple beneficial actions on lipid metabolism. A decrease in total- and LDL-cholesterol and a decreased oxidation of lipoproteins is consistently reported. Main mechanism of action could be an increase in hepatic LDLbinding eventually associated with a decreased absorption. Radical scavenging capacity may decrease atherogenicity of lipoproteins. Evidence based data from prospective controlled should be collected in order to assess the mechanism as well as the extent of action of cactus. In view of the ever growing (semi-)arid regions and the expected climatic change the intake of cactus as crop as well as dietary nutrient will gain great relevance. Key words: atherosclerosis hyperlipidemia cactus LDL-receptor prickly pear radical scavenger

potentially active nutrients and their great many properties make cactus a promising candidate to produce health promising food or supplements (Feugang et al., 2006). Mainly two desert plants recently attracted interest. The first one, Hoodia gordonii from the Kalahari desert, also called the queen of the Namic, mostly erroneously described as a cactus but in fact being a succulent plant, was used by witch doctors in South Africa to alleviate hunger and thirst. Quality control of available products is poor and heavily criticised. The other one is Opuntia, used since long by Pima Indians against diabetes mellitus, which shows an extremely high prevalence in this particular population. Discussing with local witch doctors in Arizona more than a decade ago, I recognized that Opuntia but not other cactus since long has been successfully used there as a dietary nutrient for treatment. The frequent referral and general acceptance contrast with limited data. Although since long traditionally appreciated for its pharmacological properties by a native American population but also in other parts of the world (Oh and LIm, 2006), cactus pear is still hardly recognized for that indication due to insufficient scientific information. In fact, there is a number of experimental and clinical work indicating that Opuntia may show some metabolic benefits. The overwhelming majority is dealing with the hypoglycaemic action (Frati Munari et al., 1991 ; Frai Munari et al., 1992; Roman-Ramos et al., 1995; Trejo-Gonzalez et al., 1996; Wolfram et al., 2002), while information as to lipid metabolism and in particular human is limited. It is the aim of this paper to review the current evidence of antihyperlipidemic action and potential mechanisms behind.

Introduction
The role of natural products in medicine and pharmacology is tremendously increasing in these days. On the one hand, since long drugs have been derived from various plants, on the other hand, fruits and nutrients since long used by local people for various health issues, are gaining increasing attention and also commercial interest. Among them: cacti. Cactus can be considered as an important crop in (semi-)arid regions and as a natural source of fatty acids with a variety of beneficial biological actions. The large number of

Available information History

The investigation of prehistoric diets (Danielson and Rainhard, 1998) indicates that already in this period about 6000 years ago prickly pear was widely consumed by the
29

Native American population.

Current use
Nopalitos (young sprouts) of Opuntia ficusindica are widely used in Mexico as vegetables. The Mexicans have more than 100 recipes for the preparation of nopalitos. Also in South Korea Opuntia ficus-indica is a traditional health fruit which also has been used to prevent various diseases for a long time (Han et al., 2001; Oh and Lim, 2006).

increased, most likely due to a decreased oxidation injury preserving local prostacyclin synthese activity. Ennouri and coworkers (Ennouri et al., 2005) showed a decrease in total and LDLcholesterol in rats fed prickly pear seed oil, exceptionally rich in linoleic acid, while HDL remained unchanged.

Human data
In a randomized cross-over double-blind study in 18 healthy volunteers receiving either 250 g fresh pulp of Opuntia ficus-indica or 75 mg vitamin C for 2 weeks daily there was a comparable increase in vitamins E and C. 8epi-PGF2 (an isoprostane) and malondialdehyde indicators of oxidative stress dropped by about 30 and 75%, respectively (Tresoriere et al., 2004). Opuntia streptacantha was able to decrease the area under the glucose tolerance curve and the hyperglycaemic peak (Roman-Ramos et al., 1995). It remains to be elucidated, whether the antiplatelet action reported is due to the hypolipemic and/or the antioxidant effect or a direct antiplatelet action (Wolfram et al., 2003). In one study, the edible pulp of Opuntia robusta given for 4 weeks at 250 g per day showed a significant lowering of total (p < 0.01) and LDL-cholesterol (p < 0.04), while HDL and triglycerides remained unchanged. The observed decrease in plasma, serum and urinary isoprostane (IP) 8-epi-PGF2 (Budinsky etv al., 2001) indicated that significant lowering of oxidation injury. There was a significant correlation of IP with total cholesterol and even more close to LDLcholesterol, and a negative one to HDL, the vitamin content being unable to explain the benefit. Using radioiodine labelled LDL shows a significant upregulation of LDL binding sites in men (16) determined via in-vivo gamma camera imaging. The same design used in 10 patients with isolated heterozygous familial hypercholesterolemia for 6 weeks showed a significant (p < 0.0001) increase in LDL uptake by the liver and an enhanced decay in circulating blood. Total cholesterol was lowered from a mean of 298.0 to 268.0 mg/dl (p < 0.0001) and LDL-cholesterol from 210.5 to 176.4 mg/dl (p < 0.0001). Again, triglycerides and HDL were unaffected. In another paper of the same group, mainly aimed to study the effect on glucose and lipid metabolism in non-diabetics, a decrease of total cholesterol by 12% LDL-cholesterol by

Background
Elevated lipids and lipoproteins, in particular after oxidative modification, have been shown to be a key risk factor for the development of atherosclerosis. A decrease in total cholesterol of only 1% results in a 3-percent decreased vascular risk. Thus, nutritional measures being able to modulate total- and LDL-cholesterol are of key relevance on a population base.

Experimental findings
In rats, an extract from Opuntia fuliginosa was purified and shown to normalize blood glucose. The low amount of substance administered (1 mg/kg body weight) excludes a relevant role of dietary fiber (Trejo-Gonzalez et al., 1996). On the other hand, prickly pear pectin was demonstrated to improve lipid metabolism at the hepatic receptor level (Fernandez et al., 1992; Fernandez et al., 1994). In Wistar rats (Li et al., 2005) Opuntia powder decreased total cholesterol, LDL, triglycerides, and malondialdehyde. The hypolipidemic action observed was dose-dependent. In guinea pigs, the addition of prickly pear pectin decreases LDL-cholesterol by increasing the expression of hepatic apo-B,E receptor (Fernandez et al., 1994). Fernandez et al. (Fernandez et al., 1992) showed in guinea pigs that after prickly pear pectin administration hepatic apo-B,E receptor expression (Bmax) was 60% increased and the fractional catabolic rate by 190%. In mice, Opuntia ficus-indica glycoprotein with a molecular mass of about 90 kDa has been shown to act as radical scavenger decreasing thiobarbituric acid reactive substances and increasing nitric oxide (Oh et al., 2006). It was also claimed that via blocking lipid peroxidation endothelial function may be improved by increasing relative local NO availability. Furthermore, this glycoprotein has been shown to decrease blood total cholesterol, LDLcholesterol and triglycerides. We found (unpublished data) that in rat aortic prostaglandin I2-formation was significantly

30

15% and apolipoprotein B (-9%), fibrinogen (11%), blood glucose (-11%), insulin (-11%) and uric acid (-10%) was found. HDL, apo-A 1, lipoprotein(a) and triglycerides, in contrast, remained unchanged.

References
Budinsky A, Wolfram R, Oguogho A, Efthimiou Y, Stamatopoulos Y, Sinzinger H., 2001. Regular ingestion of Opuntia robusta lowers oxidation injury. Prostagl Leukotr Essential Fatty Acids, 65, 45-50. Cho JY, Park SC, Kim TW, Kim KS, Song JC, Kim SK, Lee HM, Sung HJ, Park HJ, Song YB, Yoo ES, Lee Ch, Rhee MH., 2006. Radical scavenging and anti-inflammatory activity of extracts from Opuntia humifusa Raf. J Pharm Pharmacol; 58: 113-119. Danielson DR, Reinhard KJ., 1998. Human dental microwear caused by calcium phytoliths in prehistoric diet of the lower Pecos region, Texas. Am J Phys Anthropol; 107, 297304. Ennouri M, Fetoui H, Bourret E, Zeghal N, Attia H. 2005. Evaluation of some biological parameters of Opuntia ficus-indica. 1. Influence of a seed oil supplemented diet on rats. Bioresour Technol, 26, 189-193. Fernandez ML, Lin EC, Trejo A, McNamara DJ., 1992. Prickly pear (Opuntia sp.) pectin reverses low-density lipoprotein receptor suppression induced by a hypercholesterolemic diet in guinea pigs. J Nutr , 122, 2330-2340. Fernandez ML, Lin EC, Trejo A, McNamara DJ., 1994. Prickly pear (Opuntia sp.) pectin alters hepatic cholesterol metabolism without affecting cholesterol absorption in guinea pigs fed a hypercholesterolemic diet. J Nutr; 124, 817-824. Feugang JM, Konarksi P, Zou D, Stintzing FC, Zou C., 2006. Nutritional and medicinal use of Cactus pear (Opuntia spp.) cladodes and fruits. Front Biosci, 11, 2574-2589. Frati Munari AC, Xilotl Diaz N, Altamirano P, Ariza R., Lopez Ledesma R., 1991. The effect of two sequential doses of Opuntia streptacantha upon glycemia. Arch Invest Med Mex; 22, 333-336. Frati Munari AC, Vera Lastra O, Ariza Andraca CR., 1992. Evaluation of nopal capsules in diabetes mellitus. Gaceta Med Mex, 128, 431-436. Frati Munari AC, Fernndez-Harp JA, de la Riva H, Ariza-Andraca R, del Carmeritorres M. 2004. Effect of nopal (Opuntia sp.) on serum lipids, glycaemia and body weight. Am J Clin Nutr; 80, 668-673. Han YN, Choo Y, Lee YC, Moon YI, Kim SD, Choi JW., 2001. Monoamine oxidase B inhibitors from the fruits of Opuntia ficus-indica var. saboten. Arch Pharm Res, 24, 51-54. Li CY, Cheng XS, Cui MZ, Yan YG, 2005. Regulative effects of Opuntia powder on blood lipids in rats and its mechanism. Zhongguo Thong Yao Za Zhi, 30, 694-696. Meckes-Lozyoa M., Ibanez-Camacho R., 1989. Hypoglucaemic activity in Opuntia

Capsules
Various preparations of nopal capsules are available around the world. The application of commercial dried nopal capsules in diabetics versus placebo was not able to acutely improve lipid or glucose metabolism despite using the impractical dose of 30 capsules at a single application (Frati Munari et al., 1992; Frati Munari et al., 2004). In contrast, 500 g of broiled stems of Opuntia streptacantha given orally in 8 type II diabetics versus 6 healthy volunteers induced a significant decrease in glucose the diabetics versus the controls (Frati Munari et al., 1992). No scientific data on lipid metabolism of nopal capsules are available. Meckes-Lozoya et al. (Meckes-Lozoya aand Ibanez-Camacho, 1989) investigated the hypoglycaemic activity of Opuntia streptacantha and did not find seasonal variations concerning this effect.

Discussion
Already in 1989 Moore and coworkers (Moore, 1989) wrote that a benefit of prickly pear consumption in diabetes is clinically verifiable. Fernandez et al. (Fernandez et al., 1994) were the first to claim that prickly pear pectin intake decreases LDL-cholesterol by increasing the hepatic apo-B,E receptor expression in guinea pigs fed a hypercholesterolemic diet. The absorption, however, was unaffected. Furthermore, Opuntia humifusa for example, was reported to exert a direct scavenging activity and inhibition of reactive species generation resulting in a decreased expression of cytokines (Cho et al., 2006).

Conclusion
Consistently, in animal experiments as well as in men cactus products seem to improve lipid metabolism (lowering total- and LDLcholesterol) and decreasing the extent of lipoprotein oxidation. This review focuses on lipid action of cactus (products) in experimental animals and human. Unanimously, a beneficial effect on total- and LDL-cholesterol is reported which may be due to an increase in hepatic LDL-binding and a decreased absorption. Compositional changes and antioxidant action on lipoproteins may add a further piece to the antiatherosclerotic actions.

31

streptacantha throughout its annual cycle. Am J Chin Med; 17, 221-224. Moore M., 1989. Medicinal plants of the desert and Canyon. Museum of New Mexico Press, Santa Fe. Oh P.-H, Lim K-T, 2006. Glycoprotein (90kCa) isolated from Opuntia ficus-indica var. saboten Makino lowers plasma lipid level through scavenging of intracellular radicals in triton WR-1339-induced mice. Biol Pharm Bukk, 29, 1391-1396. Palumbo B, Ethimiou Y, Stamatopoulos J, Oguogho A, Budinsky A, Palumbo R, Sinzinger H., 2003. Prickly pear induces upregulation of liver LDL binding in familial heterozygous hypercholesterolemia. Nucl Med Review; 6, 35-39. Roman-Ramos R, Flores-Saenz JL. Alarcon-Aguilar FJ., 1995. Anti-hyperglycemic effect of some edible plants. J Ethnopharmacol, 48, 25-32. Tesoriere L, Butera D, Pintaudi AM, Allegra M, Livrea MA. 2004 Supplementation with

cactus pear (Opuntia ficus-indica) fruit decreases oxidative stress in healthy humans: a comparative study with vitamin C. Am J Clin Nutr, 80, 391-395. Trejo-Gonzalez A, Gabriel-Ortiz G, Puebla-Perez AM, Huizar-Contreras MD, MungiaMazariegos MR, Mejia-Arreguin S, Calva E., 1996. A purified extract from prickly pear cactus (Opuntia filiginosa) controls experimentally induced diabetes in rats. J Ethnopharmacol, 55, 27-33. Wolfram RM, Kritz H, Schmid P, Efthimiou Y, Stamatopoulos Y, Sinzinger H., 2002. Effect of prickly pear (Opuntia robusta) on glucose- and lipid-metabolism in nondiabetics with hyperlipidemia. Wr klin Wschr; 114, 840-846. Wolfram RM, Budinsky A, Efthimiou Y, Stamatopoulos Y, Oguogho A, Sinzinger H., 2003. Daily prickly pear consumption improves platelet function. Prostagl Leukotr Essential Fatty Acids, 69, 61-66.

32

Healthy Lipophilic Fiber from Cactus

Catherine Thimonier and Karen Jauntre
BIO SERAE Laboratoires SA 1 av. de la Preuilhe. Parc Tech. du Lauragais - 11 150 Bram France marketing@bioserae.com www.bioserae.com - info@bioserae.com; www. NeOpu ntia .com

Abstract
BIO SERAE Laboratories, French company established in 1984, has been working since the end of the 90s in the field of nutraceuticals, to develop innovative & original active ingredients. Obesity and associated disorders being one of the major issues of developed countries populations, its R&D team focused its research on weight management ingredients, and particularly on dietary fibers. After comparative studies, they chose Opuntia ficus-indica for its assets as a lipophilic fiber, and developed NeOpuntia This article will have a look on the research works made on this cactus-based ingredient, and will particularly focus on the latest clinical study conducted syndrome X related.

the stomach. There, hydrophobic interaction takes place between insoluble fibers and fat units. Primary interactions are then stabilized thanks to the soluble fibers (polysaccharides which create a fluid gel, stable at any pH). The fat/fibers interaction formed is thought to reduce fat bioavailability during digestion, and is naturally eliminated. The legal status of NeOpuntia is clear: it is a food grade ingredient made from the leaves of prickly pear cactus. Historically, the leaves have been consumed throughout Latin America and Southern Europe for centuries. NeOpuntia is thus exempted from Novel Food procedure: it means that the raw material from which the ingredient is made is considered as a traditional everyday consumption vegetable. NeOpuntia is 100% green and natural, and certified Kosher, Vegetarian and Organic (by ECOCERT S.A.S. F32600). It is made from cactus thanks to a patented gentle chemical-free production process, respectful for the plant, the environment and the human body. Major players in the nutraceuticals and in the food industries already trusted NeOpuntia and incorporated it as a branded ingredient into their weight management range of products. More than 150 finished products are available worldwide.

Introduction
NeOpuntia is a functional ingredient resulting from a long step by step development process. It is made from dehydrated leaves of cactus Opuntia ficus-indica via a patented process (PCT / FR02 / 01254). NeOpuntia is an original complex of insoluble and soluble dietary fibers which offers interesting lipophilic properties. NeOpuntia can indeed be involved to reduce fat absorption into the body, with instant efficiency starting in

Scientific background
NeOpuntia is backed by strong scientific

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research works, including various in vitro and in vivo studies. The lipophilic capacity of NeOpuntia was first tested in a dynamic in vitro model of the gastrointestinal tract. This model very closely simulates the human digestion conditions, and was conducted at TNO Nutrition and Food Research (The Netherlands). Two grams of NeOpuntia were added to a standardized meal. Various fractions were collected from all the gastro-intestinal compartments and the fatty acid composition was analyzed. This study shows that NeOpuntia absorbs 28,3% of total fatty acids during the 4-hour experiment time frame. In 2003, a pilot clinical study was conducted in partnership with the CERN (France). This double blind study was conducted on 10 volunteers (5 women and 5 men), controlled by placebo, randomised and crossed on 2 groups. It suggests that 1,6g of NeOpuntia per meal increases fat excretion on an average of 27%, compared to placebo. In 2004, a comparative TNO test with a branded chitosan showed that, under exactly the same conditions, this latest had no effect on the experiment, while NeOpuntia decreased fatty acid bio-accessibility by 28.3%. In 2005, BIO SERAE decided to renew its collaboration with the TNO institute. The efficacy of NeOpuntia to bind fat was tested by measuring the bio-accessible lipid fraction at the dosage of 2g of NeOpuntia, associated with a traditional American breakfast, intentionally rich in fats to provide the greatest effects on gastrointestinal physiology, as recommended by the FDA to test usual drugs. The results of the study confirmed lipophilic properties of NeOpuntia : associated to an American breakfast intake, NeOpuntia absorbed about 23% of total fatty acids compared to a control. This study shows a real potential interest of NeOpuntia as a food ingredient. Associated in food formula, NeOpuntia could prevent some of the fat metabolism while keeping satisfaction linked to the taste of fat.

lipid levels (HDL cholesterol, triglycerides), parameters which are linked to Syndrome X diagnosis... Metabolic Syndrome (Syndrome X), is now considered as the sum of non-pathological disorders which constitutes THE factor of cardiovascular risk, and not only blood lipid parameters. The clinical definition of Metabolic Syndrome is based on five parameters, but the presence of only 3 of them is sufficient to diagnose it. The interest of the study is thus to test NeOpuntia for its ability to regulate those 5 parameters and influence Metabolic Syndrome diagnosis. These 5 criteria as defined by the International Diabetes Federation (2005) are presented here-after, abdominal obesity being a mandatory criterion of Metabolic Syndrome: Fasting glucose > 1,00 g/l Abdominal Obesity (waist circumference) > 80cm for women, > 94cm for men Blood pressure > 130/85 mm Hg Blood triglycerides > 1,5 g/l HDL Cholesterol < 0,50 g/l for women, < 0,40 g/l for men

Methods
The objective of this clinical study was to evaluate the efficacy of NeOpuntia on blood lipid parameters. It was conducted in France between May and June 2006 on 68 females with a BMI (Body Mass Index) between 25 and 40, and diagnosed with Metabolic Syndrome as defined by the International Diabetes Federation in 2005. The study was monocentric, randomised, placebo-controlled, in parallel double blind format. Several lipid parameters were measured: changes in HDL cholesterol, serum triglycerides. At the end of the study the participants were re-evaluated for metabolic syndrome. During the 6 week study, half of the females consumed 1,6 g of NeOpuntia per meal and the other half consumed a placebo. The placebo and NeOpuntia were given in the form of capsules. All along the study, volunteers had to follow dietary advice in order to respect a balanced diet (average caloric input of 2000 kcal, lipid input limitation) and to have minimum physical activity (30 min per day). During this study, several blood lipid parameters were controlled (D0, D+14 et D+42), in particular the evolution of HDL cholesterol, triglycerides and metabolic syndrome diagnostic.

Latest clinical study, Syndrome X related

After this set of studies confirming NeOpuntias lipophilic properties and its ability to interact with fats, BIO SERAE wished to conduct a larger study designed to assess NeOpuntias ability to manage 5 risk factors typically associated with Metabolic Syndrome (Syndrome X). This latest study indeed revealed another facet of NeOpuntia and highlighted its positive role in balancing blood

34

Results
% free of m etabolic syndr

First of all, there was no observed side effect due to the consumption of NeOpuntia. The ingested dose did not imply any particular discomfort during the test : no gastric disorder nor bloated feelings were reported. NeOpuntia improved the level of good cholesterol (HDL cholesterol level), which is generally associated with a reduced cardiovascular risk. NeOpuntia indeed created a positive effect on the HDL cholesterol level of the test population, while studies show that an increase of 0,01 g/l of the HDL cholesterol level reduces cardiovascular risk by 5,5% (Lipid Research Program : the lipid research clinics coronary prevention trial results. JAMA 1984; 251: 351-374. & Circulation 1986; 74: 12171225)

40% 30% 20% 10% 0% 8% Placebo N eO puntia 39% Placebo N eO puntia

Figure 2. Women diagnosed free of Metabolic Syndrome at the end of the study Specific sub-groups (identified according to criteria such as age, glycemia or blood pressure) were even more sensitive to NeOpuntia: on a receptive population, near than 60% of women in the test group were diagnosed to be free of Metabolic Syndrome, at the end of the study. This result is statistically significant.

HDL, g/l 0.04 0.02 0 -0.02 -0.04 -0.06 NeOpuntia Placebo 0.0217 -0.047

Conclusion
To conclude, we can say that, in addition to its lipophilic properties, results indicate that NeOpuntia (patented powder of Opuntia ficus-indica) could help to balance blood lipids level and reduce cardiovascular risk due to its beneficial effects on HDL cholesterol level. This study showed that NeOpuntia could play a part in the regulation of the parameters linked to Syndrome X and influence its diagnostic, particularly regarding the HDL and triglycerides parameters. It is also interesting to note that the effects of NeOpuntia can be even more important on specific populations. This latest clinical study revealed thus another facet of NeOpuntia : beyond its lipophilic / weight management properties, NeOpuntia offers further health benefits which perfectly sticks to the current trend of the weight management market, which intends to target not only weight loss, but also health-related troubles linked to obesity (Syndrome X, for instance).

Figure 1. Evolution of HDL level (g/liter) In women 45 years or older, NeOpuntia caused a large improvement by increasing levels of good HDL-cholesterol and decreasing levels of harmful triglycerides. The NeOpuntia group of this specific subdivision benefited from a favourable evolution of their HDL level (significant difference between the 2 groups evolution: p = 0.029). This effect has been reinforced by a positive evolution of triglycerides according to a p close to the significant trend. And, at the end of the study, near than 40% of women in the NeOpuntia group was no longer diagnosed with Metabolic syndrome (Syndrome X).

35

Beneficial Effect Of Opuntia Ficus Indica Seeds And Oil On Animal Health
Monia Ennouri
Alimentary Analysis Unit, National Engineering School of Sfax, BPW 3038, Sfax, Tunisia Higher Institute for the Applied Sciences and Technology of Mahdia, FSEG Sidi Messaoud 5111, Mahdia, Tunisia E-mail address: Monia.Ennouri@enis.rnu.tn

Abstract
The purpose our investigation concern the evaluation of the effects of diets enriched with cactus pear oil (CPO) or seeds (CPS) on serum and liver lipid parameters compared to those of adult rats submitted to standard diet were tested. Male rats were divided into three groups, the first group represented control group, fed with standard diet, the second group was fed with control diet supplemented with CPO (2.5%, wt/wt) and the third group fed control diet supplemented with CPS (33%, wt/wt), for nine weeks. Feed intake and body weight of rats were measured every two days. Cholesterol, HDL and triglycerides levels were determined by enzymatic methods. Liver and serum lipid extracts were analysed for their fatty acid composition for the three groups of rats. The tested diets decreased significantly the atherogenic index compared to control diet, whereas serum cholesterol level was only reduced by the supplementation with CPO diet. No variations in serum lipids were observed among the groups, whereas liver lipids showed slight variations. Accordingly, these results indicated that the supplementation with CPO or CPS could be effective in decreasing the atherogenic risk factors in rats Key words: Opuntia ficus-indica seeds; Fatty acids profile; Liver; Serum; Cholesterol

the structure of their glucuronoxylans has been identified by Habibi, Mahrouz, and Vignon (2002). The seeds of Opuntia ficus-indica (cv. Gigante) contain a storage protein having a molecular mass of 6.5 kDa isolated and characterized by Uchoa, Souza, Zarate, Gomez-Filho and Campos (1998). The cactus pear seeds proteins are rich in mineral and sulfur amino acids, such as methionine and cysteine which represent nearly twice the FAO/WHO (FAO/WHO, 1973) recommended requirements for human. Seeds composition of cactus pear fruits during the maturation period has been studied by Coskuner and Tekin (2003). Their nutritive value has been studied by Sawaya, Khalil, and Al-Mohammad (1983). The cactus pear seed oil composition and its chemical characteristics were investigated by Sawaya and Khan (1982), and by Salvo, Galati, Lo Curto and Tripodo (2002). Dietary fatty acids have a significant effect on plasma cholesterol and levels of lipoproteins, which are linked to the incidence of coronary heart disease (CHD) (Hu, Mason & Willer, 2001; Yu, Derr, Etherton & Kris-Etherton, 1995). Recently, the use of functional food and/or nutraceuticals has increased due to their beneficial effects on human health (Dubick, 1986). Among Mediterranean plants, prickly pear seeds and oil exhibit hypoglycemic and hypocholesterolemic effects (Ennouri, Fetoui, Bourret, Zeghal & Attia, 2006a, Ennouri et al., 2006b) due probably to the fatty acid composition of the prickly pear seeds oil (Ennouri, Bourret, Mondolot & Attia, 2005). Changes in the fatty composition of several tissues such as liver and serum could be attributed to the type of fat ingested in the diet. Therefore, the aim of the current study was to evaluate the effect of diet supplemented with cactus pear oil (CPO) or cactus pear seeds (CPS) comparatively to control diet, given to

Introduction
Cactus pear (Opuntia ficus-indica) grows throughout Tunisia and the fruits are consumed exclusively as fresh fruit. The seeds are rich in oil, known for its content in polyunsaturated fatty acids. The seeds are made of two different tissues, the endosperm and the pericarp in the relative proportion of 1:9 respectively. Analysis of the main constituents of prickly pear seeds showed a significant amount of polysaccharides, cellulose and hemicelluloses,

36

adult rats, on body weights, lipid parameters in serum and fatty acids profile in their serum and liver.

Results Chemical composition of diets

Table 1 shows the chemical composition of fed diets substituted with oil (CPO) or seed powder (CPS) and the control diet. The CPS diet was rich in lipids and fiber compared to control. The CPO diet was also rich in lipids compared to control.

accumulation of lipids in liver. As the fiber content was the only major variable in the ingredients of CPO and CPS diets (Table 1), the supplementation of diet with whole cactus pear seeds was better in lowering the lipid levels of hepatic tissues.

Serum and liver lipid profiles

In previous study we have demonstrated that cactus pear seed oil was rich in oleic (C18:1) and linoleic (C18:2) acids (16.7% and 70.3% respectively), which representing 87% of the total fatty acids (Table 4) (Ennouri et al. 2005). The type of fat ingested in the diet could provoked changes in the fatty composition of tissues, which is why we have tried to study the fatty acid composition of serum and liver lipid extracts on rats after enrichment of diets with CPO and CPS. The major fatty acids in serum as well as on liver of all groups were arachidonic, palmitic, linoleic, oleic, and stearic acids (Table 5, Table 6). Netherless the fatty acid analysis in liver showed that, in CPO group the significant increase in oleic acid level resulted in an increase of MUFA. A reduction in linolenic acid in this group provoked a significant decrease in PUFA. But in CPS group, the level of stearic acid was increased by 27%, while linolenic acid was decreased by 52%. The levels of docosahexaenoic, docosatetraenoic, arachidonic and linoleic acids representing 45% of total liver fatty acids; were similar in CPO and CPS compared to control group (Table 6). In the CPS group a decrease by 8% of PUFA in serum and liver was observed compared to control. Indeed, the ratio of PUFA to SFA decreased significantly in CPS group (Table 5, Table 6). CPO and CPS diets contained the same amount and quality of cactus pear seed oil. It is known that oleic (C18:1) and linoleic (C18:2) acids have beneficial health effects, including alleviating cardiovascular complaints, inflammatory conditions, heart diseases, atherosclerosis, autoimmune disorder, diabetes and other diseases (Hegsted, McCrancy, Myers & Stare, 1965). Linoleic acid is an essential fatty acid and a precursor of arachidonic acid biosynthesis, the substrate for eicosanoid synthesis. According to Keys, Anderson and Grande (1957), linoleic acid has hypocholesterolemic effects.

Feed intake and Body weight of rats

A significantly lower average gain of body weight in the CPS group was obtained during the nine weeks of treatment despite a greater feed consumption than those of the control and CPO groups. In the CPO group, no significant body weight gain was obtained compared to control (Table 2).

Effect of diet substitution on plasma cholesterol and triglyceride concentrations

The CPO group had a significant reduction in total cholesterol and the ratio of HDL cholesterol to total cholesterol was significantly higher than that of the control group (Table 3). The last parameter was negatively correlated with the risk of coronary heart disease, as reported by Barter and Rye (1996). An increase by 93% of serum triglyceride level was noted in CPO group, this increase may be explained by the rise of serum lipid level as reported in our results. In CPS group the ratio of HDL cholesterol to total cholesterol increased significantly compared to control, since HDL cholesterol was increased, while serum triglyceride level decreased by 53% compared to control group. The atherogenic index was significantly lower in CPO and CPS than the control group. This result was in agreement with Davignon and Cohn (1996) who have indicated that a decrease of serum triglyceride level was associated with lower risk of coronary disease.

Serum and liver lipid contents

The lipid content of serum increased significantly in the CPO and CPS groups compared to control (Fig.2 A) whereas in the CPS group a significant decrease in liver lipid content was noted (Fig.2 B). The reduction in the level of liver total lipids in CPS group paralleled that of serum triglycerides. The liver weight was greater in CPO group than in the control group, it was probably due to the

Conclusion
Our studies demonstrated for the first time that the enrichment of diet with CPS had a very pronounced hypolipidemic effect as compared to the CPO diet. It could significantly decrease the levels of triglycerides in serum and total

37

lipids in liver. There were no major variations for the fatty acid composition of liver and serum extracts. More studies are needed to explain the potential hypocholesterolemic and

hypolipidemic effects of Opuntia ficus-indica seeds and oil extract on hypercholesterolemic and other pathologies on rats.

Table 1. Chemical composition of fed diets (g/100g d.m.)a Ingredients Lipids Protein Ash Cellulosic polysaccharides Starch
a

Control 3.37 0.17

x

CPO 5.90 0.10

x

CPS 5.87 0.36y 11.95 1.12x 3.53 0.11x 23.41 1.25y 8.30 1.32x

15.23 1.35x 4.63 0.09x 5.12 1.06x 11.75 1.86x

14.68 0.52x 4.50 0.15x 5.20 0.09x 11.63 1.50x

Each value is the mean of three observations standard error. Figures having different letters (superscribed) in the same row are significantly different (P 0.05).

Table 2. Initial body weight, body weight gain and feed intake in control, CPO and CPS groups a after 9 weeks of treatment . Parameters Initial weight (g) Average weight gain (g/rat) Average feed intake (g/day) Control 122.35 1.61 20.7 4.6x
x

CPO 123.7 0.9

x

CPS 123.8 0.7x 93.7 15.5y 25.2 3.7z

143.3 31.2x

141.8 22.6x 18.6 1.5y

a Each value is the mean of seven rats standard error. Figures having different letters (superscribed) in the same row are significantly different (P 0.05).

Table 3. Lipid parameters in serum of rats in CPO, CPS and control groups. Parameters Cholesterol (g/l) HDL cholesterol (g/l) HDL cholesterol /Total Cholesterol (%) Triglycerides (g/l) Athero. Index* Control 0.66 0.03x 0.36 0.03x 0.55 0.06x 0.717 0.079x 0.822 0.204x CPO 0.43 0.06y 0.35 0.08x 0.81 0.20y 1.384 0.182y 0.505 0.205y CPS 0.76 0.03y 0.48 0.03y 0.64 0.02z 0.333 0.075z 0.564 0.071y

a Each value is the mean of seven rats standard error. Figures having different letters (superscribed) in the same row are significantly different (P 0.05). *Athero. index (atherogenic index)= (total cholesterol - HDLcholesterol)/ HDLcholesterol (Deguchi & Ogata, 1991).

38

Table 4. Fatty acid composition of prickly pear seed oil (g / 100 g of total fatty acid) (Ennouri et al., 2005) Fatty acid Lauric C12:0 Myristic C14:0 Palmitic C16:0 Palmitoleic C16:1 Stearic C18:0 Oleic C18:1 Linoleic C18:2 SFA
c d

Opuntia ficus indica b b 9.320.19 1.420.01 3.110.04 16.770.47 70.290.60 12.430.12 18.190.47 70.290.60

MFA

PFAe

a Each value is the mean of three observations standard error. b Not detected. c Saturated fatty acids. d Monounsaturated fatty acids. e Polyunsaturated fatty acids.

Table 5. Serum fatty acid composition of rats in CPO, CPS and control groups (g/100 g of total a fatty acid) . Fatty acid SFA Myristic acid (C14:0) Palmitic acid (C16:0) Stearic acid (C18:0) MUFA Palmitoleic acid (C16:1) Oleic acid (C18:1) PUFA Linoleic acid (C18:2) Linolenic acid (C18:3) Eicosatrienoic acid (C20:3) Arachidonic acid (C20:4) Eicosapentaenoic acid (C20:5) Docosatetraenoic acid (C22:4) Docosahexaenoic acid (C22:6) PUFA/SFA Control 32.0 5.75
x

CPO 32.440 0.878 0.409 0.002y 19.98 0.84y 11.04 0.01y 14.75 1.02y 0.7 0.50y 13.49 0.71a 52.7 2.35x 22.71 1.43y 0.73 0.01y 0.92 0.36y 25.25 2.36x 0.16 0.07x 0.82 0.02y 1.109 0.516y 1.626 0.108x
x

CPS 34.891 0.236y 0.501 0.049z 20.3 1.02y 12.69 0.68z 16.67 2.46x 1.26 0.03y 13.1 2.41 47.6 2.25y 17.18 0.45z 0.95 0.18z 0.71 0.17y 25.72 2.99x 0.33 0.10x 0.77 0.03x 2.08 0.42z 1.36 0.05y

0.45 0.00x 21.2 0.31x 9.23 0.35x 18.67 0.7x 1.98 0.53x 15.81 0.47x 52.01 1.36x 24.02 1.23x 0.80 0.04x 0.26 0.01x 23.771 0.86x 0.16 0.07x 0.71 0.02x 2.130 0.226x 1.613 0.027x

a Each value is the mean of seven rats standard error. Figures having different letters (superscribed) in the same row are significantly different (P 0.05).

39

Table 6. Liver fatty acids composition of rats in CPO, CPS and control groups (g/100 g of total a fatty acid) . Fatty acid SFA Myristic acid (C14:0) Palmitic acid (C16:0) Stearic acid (C18:0) MUFA Palmitoleic acid (C16:1) Oleic acid (C18:1) PUFA Linoleic acid (C18:2) Linolenic acid (C18:3) Eicosatrienoic acid (C20:3) Arachidonic acid (C20:4) Eicosapentaenoic acid (C20:5) Docosatetraenoic acid (C22:4) Docosahexaenoic acid (C22:6) PUFA/SFA Control 38.88 2.99
x

CPO 36.53 0.32

x

CPS 44.9 3.82z 0.23 0.04x 20.66 1.69x 22.90 2.09y 10.12 0.91x 0.72 0.07x 7.74 1.26x 46.43 2.43y 14.43 1.69y 0.28 0.01y 0.42 0.03x 24.0 2.15x 0.11 0.04x 1.15 1.06x 4.21 0.38x 1.04 0.14y

0.26 0.06x 18.93 0.64x 18.0 2.64x 10.6 2.56x 0.77 0.14x 8.62 2.60x 50.1 0.46x 17.51 1.70x 0.48 0.07x 0.34 0.11x 23.42 1.04x 0.43 0.32x 1.29 0.1.21x 4.76 0.13x 1.29 0.08x

0.51 0.05y 19.44 0.18x 15.24 0.1y 15.0 0.46y 0.97 0.34x 13.6 0.34y 48.18 0.47y 17.33 0.48x 0.25 0.17y 0.58 0.02y 22.46 0.16x 0.06 0.05y 1.52 0.34x 4.11 0.54x 1.32 0.02x

a Each value is the mean of seven rats standard error. Figures having different letters (superscribed) in the same row are significantly different (P 0.05).

30 Lipid content (mg/ml) 25 20 15 10 5 0

A
*** (7) (7) ** (7)

Control CPO CPS

Serum

Fig. 2. Effect of diet supplemented with CPO or CPS on serum lipid content (A) and liver lipid content (B) compared to control.
Control CPO CPS

200 Lipid content (mg/g) 160 120 80 40 0

B
(7)

** (7)

Values are given as means SEM. Significant differences between the treated and controls groups ***: P0.001 and **: P0.01 The number of rats studied are shown between brackets.

*** (7)

liver

40

References
P.J., Rye, K.A., 1996. High density lipoproteins and coronary heart disease. Atherosclerosis 121, 1-12. Coskuner Y., & Tekin A., 2003. Monitoring of seed composition of prickly pear (Opuntia ficusindica L) fruits during maturation period. J. Sci. Food Agri. 83 (8), 846-849. Davignon, J., and Cohn, J.S., 1996. Triglycerides: A risk factor for coronary heart disease. Atherosclerosis 124 (suppl), S57-S64. Deguchi, Y, and Ogata, A., 1991. Relationship between serum selenium concentration and atherogenic index in Japanese adults. Tohoku J. Exp. Med. 165, 24751. Dubick, A.M., 1986. Historical perspectives on the use of herbal preparations to promote health. J. Nut. 116, 1348-1354. Ennouri, M., Bourret, E., Mondolot, L., Attia, H., 2005. Fatty acid composition and rheological behaviour of prickly pear seed oils. Food Chem. 93, 431-437. Ennouri M., Fetoui H., Bourret E., Zeghal N., Attia H. 2006a. Evaluation of some biological parameters of Opuntia ficus-indica: 1Influence of oil supplemented diet on rats. Biores. Techn. 97, 1382-1386. Ennouri M., Fetoui H., Bourret E., Zeghal N., Guermazi F., Attia H. 2006b. Evaluation of some biological parameters of Opuntia ficus indica: 2-Influence of seed supplemented diet on rats. Biores. Techn. 97, 2136-2140. FAO/WHO, 1973. Energy and protein requirements. FAO Nutritional Meeting Rept. Ser No 52, WHO technical Rept. Ser No 522. Rome: Food and Agriculture Organisation of the United Nations. Habibi, Y., Mahrouz, M., & Vignon, M.R. 2002. Isolation and structure of D-xylans from Barter,

pericarp seeds of Opuntia ficus-indica prickly pear fruits. Carb. Res. 337, 1593-1598. Hegsted, D.M., McCrancy, R.B., Myers, M.L., Stare, F.M., 1965. Quantitative effects of dietary fat on serum cholesterol in man. Am. J. Clin. Nutr. 17, 281-295. Hu, F.B., Mason, J.E., & Willet, W.C. 2001. Types of dietary fat and risk of coronary heart disease: a critical review. J.Am. coll. Nut. 20, 5-19. Keys, A., Anderson, J.T., Grande, F., 1957. Prediction of serum cholesterol response of man to change in fats in the diet. Lancet. 2, 959-966. Salvo, F., Galati, E.M., Lo Curto S., Tripodo M.M. 2002. Study on the chemical characterization of lipid composition of Opuntia ficus-indica L. seed oil. Rivista Italiana delle Sostanze Grasse, 79 (11), 395398. Sawaya, W.N., & Khan, P. 1982. Chemical characterization of prickly pear seed oil, Opuntia ficus indica. J. Food Sci., 47, 20602061. Sawaya, W.N., Khalil, J.K., Al-Mohammad, M.M., 1983. Nutritive value of prickly pear seeds, Opuntia ficus-indica. Plant Foods Hum Nutr. 33, 91-97. Uchoa, A.F., Souza, P.A.S., Zarate, R.M.I., GomezFilho, E., & Campos, F.A.P., 1998. Isolation and characterization of a reserve protein from the seeds of Opuntia ficus-indica. Braz. J. Med. Biol. Res., 31, 757-761. Yu, S., Derr, J.E., Etherton, T.D., & Kris-Etherton, P.M. 1995. Plasma cholesterol-predictive equations demonstrate that stearic acid is neutral and monounsaturated fatty acids are hypocholesterolemic. Am .J. Clin. Nut., 61, 1129-1139.

41

Preventive Role of Cactus (Opuntia ficus-indica) Cladodes on the Biochemical and Immunolgical Toxicity of Chlorpyrifos in Mice
Saida Ncibi1*, Dalel Brahim 1, Amira Akacha1, Mohamed Naceur Krifi 2, Sami Souid1 and Lazhar Zourgui1
(1) Unit de Recherche de Biochimie Macromolculaire et Gntique (BMG), Facult des sciences de Gafsa, cit Zarroug 2112 Gafsa, Tunisie. (2) Laboratoire dImmunobiotechnologie et de Biovalorisation, Institut Suprieur des Technologies Mdicales de Tunis, 9 rue Prof. Zouhar Essafi, 1006 Tunis, Tunisie

Abstract
Extract of Cactus (Opuntia ficus-indica) cladode was given alone or simultaneously with a toxic chlorpyrifos dose to Balb/c mice and was evaluated for its ability to restore damages induced by this organophosphorus insecticide. The experimental approach consisted of six treatments of six mice: one group have received chlorpyrifos at 10 mg/kg (bw) combined with extract of cactus cladodes 100 mg/kg (bw). Another group have received chlorpyrifos at 150 mg/kg (bw) combined with extract of cactus cladodes 1.5 g/kg (bw). Two groups have received chlorpyrifos alone at 10 mg/kg and 150 mg/kg (bw) respectively. The rest of groups are control groups, (water, and 5 g/kg bw of extract of cactus cladodes). Twenty four hours after treatment, Liver damage was evaluated by the liver weight and the measure of some enzymes in serum. Immunotoxicity was estimated by the count of stem cells in the bone marrow and the weight of thymus. An increase in enzyme levels and liver weight, a decrease of the stem cells number in the bone marrow and the thymus weight were noticed in only chlorpyrifos treated animals compared with the control animals. For groups treated with chlorpyrifos and cactus, there was a recovery in all parameters, so this extract contains active components that reduce damages caused by this insecticide. Key words: cactus, chlorpyrifos, liver, thymus, bone marrow

MA 1996; Knishinsky 1971; Tesoriere et al. 2004). Opuntia ficus-indica belongs to the Cactaceae family, this plant grows wild in arid and semi-arid regions, where the production of more succulent food plants is severely limited. Many uses of cladodes and cactus pear fruit are reported (Hoffmann 1980). Cactus pear contains pectin, carotenes, betalains, ascorbic acid, quercetina and quercetin derivatives all of which have antioxidant activity (Tesoriere 2004; Wang 1988; nanjing 1976). Cladodes are consumed as fresh vegetables or added to casseroles (Hamdi, 1997; Saenz 2000). Cladodes have been investigated as possible treatment for gastritis, hyperglycemia, arteriosclerosis, diabetes and prostatic hypertrophy (Frati, Jimenez & Ariza, 1990; Hegwood, 1990; Palevitch, Earon & Levin, 1993). Chlorpyrifos (C9H11Cl3NO3PS) is the main element of many organophosphorous insecticides. In agriculture, these products are widely used (Jacobsen et al. 2004). This use is recognized as an economic approach to control pests quality and quantity, but at the same time it can be harmful for many species after consumption of its residues in agricultural products (Rastrelli et al. 2002; Colosio et al. 2005).This insecticide causes many damages for human and animal health. Its effects on nervous system are well known through the inhibition of the acetylcholinesterase enzyme, which plays an important role in neurotransmission at cholinergic synapses by rapid hydrolysis of neurotransmitter acetyhlcholine in choline and acetate (Garcia et al. 2005). However its immunotoxicity and hepatotoxicity are not very explored. Alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) are liver specific enzymes. Measure of their activities is usually carried out to check up the hepatotoxicity of different chemical agents (drugs, metals, pesticides). Both relative organ weight of liver and increase in transaminase
42

1. Introduction
Natural product and health foods have recently received a lot of attention both by health professionals and the common population for improving overall well-being, as well as in the prevention of diseases. Cactus (Opuntia) has been used for many years as a common vegetable and as medicine by the native Americans and Mexicans (Cornett 2000, Kay

and phosphatase alkaline (AkP) activities can indicate tissue damage in this organ (Prati et al. 2002). For the immunotoxicity, it can be investigated by the count of stem cells in the bone marrow and the relative weight of thymus which are elements of the primary immune system Galloway and Handy 2003). In this study, our aims were twofold: (1): to investigate changes induced by the CPF administration to SWISS mice in serum biochemical (ALAT, ASAT, and AkP), the relative body weight of liver, number of stem cells in bone marrow and the relative body weight of thymus and (2) to evaluate the safety and efficacy of cactus stem extract to ameliorate the deleterious effects of this insecticide.

and added to the granulated, mixed and homogenized and made into pellets. After 3 weeks of treatment, blood samples were collected from the retro orbital sinus in tubes containing heparin as anti-coagulant in order to determine enzymes activities in serum by spectrophotometric techniques. All animals were sacrified by cervical dislocation; thymus and liver were dissected out to check their relative organ weights (ROW). Also, the bone marrow was aspirated from the femur to do the count of stem cells.

2.3. Statistical analysis

All data were analysed statistically using students t-test, statement of significance were based on probability of p to 0.05.

2. Materials and methods

2.1. Chemicals
Chlorpyrifos CPF was purchased as an agricultural product used in our country (Duracid) from STEC society. Marker enzymes of liver damage, alkaline phosphatase (AkP), alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) were measured by standard kits provided by BioMagreb.

3. Results
Concerning lethality essays, LC50 of chlorpyrifos was confirmed, it is about 150mg/kg bw. No death occurred in groups treated with cactus alone or cactus with CPF (groups 5 and 6) indicating that this natural plant is not toxic (Viala 1980). As shown in table 1, results of serum biochemical analysis showed an increase of the biochemical parameters of interest in groups treated only with CPF (groups 2 and 4) compared to the control group (group 1), which indicates a liver damage. However, treatment by cactus restores this increase in groups treated by CPF and cactus (groups 3 and 5) in values similar to normal. The relative liver weight increases in CPF treated groups (groups 2 and 4) compared to the control (group 1). This trend was not seen in groups 3 and 5 (table 1), which can confirm the CPF toxicity (Mahaboob et al. 2005) and the protective role of cactus. The immunotoxicity was investigated through thymus and the bone marrow. The weight of thymus diminished in group 2, in group 4, this organ was totally deleted. However, it was obvious that the thymus atrophy was minimized in groups 3 and 5 (figure 1). For the bone marrow, action of this insecticide was noticed essentially in the group 4: the number of stem cells of the group 4 animals femur decreased compared with none treated. This decrease is less important when cactus is administrated (group 5) (figure 2). Thus, these results confirm immunotoxicity of CPF (Blakely et al. 1999) and allow us to say that cactus stem extract is also reliable counter harmful action of CPF.

2.2. Animals
In this study, 36 male SWISS mices (SEXUAL, St Doulchard, France) were used. Their weight was about 25g and they were 4 to 5 weeks old. Animals were housed in plastic cages, fed ad libitum and allowed to adjust to the new environment for 2 weeks time before starting the experiment. Then, they were divided into 6 treatment groups, each of six mice as follow: 1. Mice given granulated chow and drinking water ad libitum= control group (Cont. G1) 2. Mice given CPF at 10mg/kg (bw) (G2) 3. Mice given 10mg/kg (b.w) CPF with 100mg/kg (b.w) cactus (G3) 4. Mice given 150 mg/kg (b.w) CPF (G4) 5. Mice given 150 mg/kg (b.w) CPF with 1.5 g/kg cactus (G5) 6. Mice given 5g/kg cactus (G6) Group 6 was chosen to test the toxicity of cactus through lethality of mice parameter. Both CPF and cactus were dissolved in water

43

Thymus relative weight in %

0,35 0,3 0,25 0,2 0,15 0,1 0,05 0 Groupe 1 Groupe 2 Groupe 3 Groupe 4 Groupe 5

Groups

Figure.1: Liver weight following chlorpyrifos exposures and the protective role of cactus stem juice. Values are means from 6 mice. * Significantly different p0.05.
Stem cells number in million
10 9 8 7 6 5 4 3 2 1 0 Groupe 1

Groupe 4

Groupe 5

Groups

Figure 2: Number of cells in the bone marrow following chlorpyrifos exposures and the protective role of cactus stem juice. Values are means from 6 mice. * Significantly different p0.05.

4. Discussion
In the present study, both lethal LC50 and the th sub-lethal (1/10 of LC50: almost 10mg/kg) concentrations were studied. For the LC50 treatment (group 4), mortality was observed and touched half of animals, so we confirm results found by other studies (McCollister et al. 1995) With sub-lethal concentration, no mortality was observed during the experiment tenure. However, an increase in liver specific enzymes was noticed. Elevation in these

transaminases indicates the utilisation of amino acids for the oxidation or for glucogenesis and is used to determine liver damage (Philip et al.. 1995). Also, the elevation in alkaline phosphatase suggests an increase in the lysosomal mobilization and cell necrosis due to pesticide toxicity. The result of this study show increased activities of transaminases and alkaline phosphatase in serum which are conventional indicators of liver injury (Rao 2006)]: when the liver cell membrane is damaged, varieties of enzymes

44

normally located on the cytosol are released into the blood stream. Also, the increase of liver weight can be considered as an indicator of liver damage (Amacher et al. 1998). So we can conclude that this organophosphorus insecticide causes liver damage in CPF treated animals with control animals. The same biochemical parameters tests demonstrated the beneficial role of the cactus stem extract addition, so this later was able to protect liver against CPF damages. Polyphenols and flavonoides are known by their ability to increase formation and excretion of detoxified metabolites resulting from xenobiotic metabolism (Higdon and Frei 2003) and thus they can protect liver. In this field, cactus stem is recognised by its content on flavonoides such as Quercetin, Taxifolin, kaempferol.etc], to which the liver protective role can be assigned. Concerning immune system, the diminution of stem cells number is a firm indicator of its destruction (Miyan et al. 1998), since the bone marrow is the principal source of different kind of the immune cells. CPF at the LC50 causes a decrease of these cells number, thus we can say that this product gravely affects this very essential system to survive. The magnificent thing is that cactus restores this alteration even for a light percentage (about 50%). As well as the bone marrow, thymus can express the immune system status (Jacobsen and al; 2004), according to our results, this organ is more sensible to this product since it can be affected even at the sub-lethal dose. Again, cactus minimizes this atrophy. So we can conclude that this plant is able to protect thymus from CPF toxicity. According to Kim J.D (Kim and al. 2006), this protection can be attributed to flavonols which can modulate immune response. Acknowledgements. This research was funded by the Tunisian Ministry of Scientific Research and Technology through the Research Unit of Macromolecular Biochemistry and Genetics, Faculty of Sciences of Gafsa.

5. References
Amacher D.E, Schomaker S.J and Burkhardt J.E. The Relationship Among Microsomal, enzyme Induction, Liver Weight and Histological Change in Rat Toxicology Studies. Food and Chemical Toxicology. 36 9-10 (1998) 831-839 Blakely B.R, Yole M.J, Brousseau P, Boermans H,

Fournier M. Effect of chlorpyrifos on immune function in rats. Vet Hum Toxicol. 3 (1999) 140-4. Cornett J: How Indians used desert plants. Nature trails Press; 2000. Frati, A. C., Jimenez, E., & Ariza, C.R., 1990. Hypoglycemic effect of Opuntia ficus-indica in non insulin-dependent diabetes Mellitus patients. Phytotherapy Reasearch, 4, 195197. Hamdi, M., 1997. Prickly pear cladodes and fruits as a potential raw material for the industries; Bioprocess Engineering, 17, 387-391. Hegwood, D.A., 1990. Human health discoveries with Opuntia sp. (prickly pear). Horticultural Science, 25,1515-1516 Higdon J.V and Frei B., 2003. Antioxidant activity of tea polyphenols: Health Effects, Metabolism and Antioxidant functions. Critical reviews in Food Science end Nutrition. 43, 1, 89-143. Hoffmann, W., 1990. The many uses of prickly pear (Opuntia Mill.) in Peru and Mexico. Plant Research Development, 12, 58-68. Jacobsen H, stergaard G, Lam H.R, Poulsen M.E, Frandsen H, Ladefoged O and Meyer O., 2004. Repeated dose 28-day oral toxicity study in Wistar rats with a mixture of five pesticides often found as residues in food: alphacypermethrin, bromopropylate, carbendazim, chlorpyrifos and mancozeb. Food and Chemical Toxicology, 42, 8, 1269-1277. Kay MA, 1996. Healing with plants in the America, and Mexican west. The University of Arizona Press. Kim J.D, Liu L, Guo W and Meydani M., 2006. Chemical structure of flavonols in relation to modulation of angiogenesis and immune-endothelial cell adhesion.The Journal of Nutritional Biochemistry. 17, 3, 165-176. Knishinsky R, 1971. Prickly pear cactus medicine. Healing Arts Press. Rochester, Vermont. Mahaboob S.K, Sobtib R.C, Katariaa L., 2005. Pesticide-induced alteration in mice hepato-oxidative status and protective effects of black tea extract. Clinica Chimica Acta 358, 131138. McCollister S.B, Kociba R.J, Humiston C.G,. McCollister D.D and Gehring P.J., 1974. Studies of the acute and long-term oral toxicity of chlorpyrifos (o,o-diethyl-O-(3,5,6trichloro-2-pyridyl) phosphorothioate). Food and Cosmetics Toxicology. 121, 45-61. Miyan J.A, Broome C.S and Afan A.M., 1998. Coordinated host defense through an integration of the neural, immune and haemopoietic systems. Domestic Animal Endocrinology. 15, 5, 297-304. Nanjing Pharmacologic College: Zhong Cao Yao Xue Jiangsu People Press; 1976: 681-682. Palevitch, D., Earon, G, & Lezvin, I., 1993.

45

treatment of beningn prodtatic hypertrophy with Opuntia ficus-indica (L); Miller. Journal Herbs Spices Medical Plants, 2, 45-49. Philip G.H, Reddy P.M, Sridevi G., 1995. Cypermethrin induced in vivo alterations in the carbohydrate of freshwater fish Labeo rohita. Ecotoxicol. Environ. Saf. 31,173178. Rao J.V. Toxic effects of novel organophosphorus insecticide (RPR-V) on certain biochemical parameters of euryhaline fish, Oreochromis mossambicus. Pesticide biochemistry and physiology. In press, available on line 28 February 2006. Saenz, C., Mecklenburg, P., Estevez, A. M., & Sepulveda, E., 1996. Natural liquid sweetener from cactus pear: obtention and

characterisitics. Acta Horticulture, 438, 135-138. Tesoriere L, Butera D, Pintaudi M, Allegra M, Livera MA, 2004; Supplementation with cactus pear (Opuntia ficus-indica) fruit decreases oxidative stress in healthy humans: a comparative study with Vit.C, Am J Clin Nutr , 80, 391-395. Toxicon, 47, 567-574. Viala A. Dfinitions, domaines de la toxicologie, notions sur la toxicit: in Elments de toxicologie. Tec & Doc, Lavoisier, 11 rue LavoisierF75384, Paris Cedex 08, Editions mdicales internationales, Alle de la croixBosse. Wang PZ: Chinese Medicine Surgery. Ancinet Chinese Medicine Press: 1988, 164-183.

Table 1: Effect of cactus stem extract and its protective role against hepatotoxicity in male SWISS mice treated at 10mg/kg (b.w) and 150 mg/kg (b.w) with CPF Control (G1) Cactus 1.5g/kg (G6) (G2) CPF 10mg/kg 150mg/kg (G4) CPF + cactus 10mg/kg CPF + 100 mg/kg cactus (G3) 150mg/kg CPF +1.5 g/kg cactus (G5) ALAT (UI/ml) ASAT (UI/ml) LDH (UI/l) PAL (UI/l) 13713 12826 1644384 9614 13618 1255 1703225 1036 21017* 23653* 3156161* 20649* 34917* 47593* 3993123* 28328* 16013 23653 2293407 12110 23010* 39143* 311640* 19921*

Values are means (n=6) * Significantly different from the control (G1) p0.05.

46

Cactus Pear: A Natural Product In Cancer Chemoprevention

Changping Zou
Associate Professor Department of OB/Gyn, Gynecologic Oncology University of Connecticut Health Center, USA Email: zou@uchc.edu Remarkable progress has been made over the past two decades in cancer prevention and chemoprevention as well as understanding the molecular and cellular mechanisms of carcinogenesis and cancer progression, as a result, the incidences of certain types of cancer have decreased significantly in US due to prevention modalities and improved screening technology. However, the incidence and mortality rates of ovarian cancer have remained essentially unchanged. Nonetheless, the development of effective and safe agents for prevention and treatment of cancer remains slow, inefficient, and costly, with little to offer the high-risk population for primary prevention and cancer survivors to prevent cancer recurrence. The key to effective chemoprevention is the identification of a chemopreventive agent(s) that can effectively inhibit cancer development with little toxic side effects. Chemoprevention by using fruit and vegetables as well as isolated compounds from food and natural products has been carried out in recent years in the United States. Medical benefits from plants have been recognized for centuries. Herbs have been used by Native American and Chinese medicine for thousands of years to cure diseases and heal wounds. Also as a rule, herbs and natural products lack much of the toxicity that is present in synthetic chemicals thus, enhancing their appeal for long term preventive strategies. Cactus (Opuntia) has been used for many years as a common vegetable and as medicine by the Native Americans and Mexicans. Cactus pear contains pectin, carotenes, betalains, ascorbic acid, quercetina and quercetin derivatives all of which have antioxidant activity. In Chinese medicine, cactus fruit is considered a weak poison and used as a detoxification agent for snake bites, it also in treatment of inflammation, pain. It has been noted that Native Americans have a lower cancer rate when compared to white and African Americans, it might be in part by using antioxidant dietary supplement such as cactus pear and nopale. We tested the ability of cactus pear fruit mixture extract in suppressing carcinogenesis in cultured human cancer cell lines in vitro and an animal model in vivo. Our results show that the cactus pear mix extract inhibited growth of ovarian, bladder, and cervical cancer cells in vitro and suppressed tumor growth in animal inoculated with ovarian cancer cells. Cactus products inhibited cancer cell growth with concentrations as low as 5%; cell cycle was also affected at this concentration with an increase in G1 phase. Apoptosis was observed at a higher concentration of 10% and 25%. We also compared cactus with the chemopreventive agent 4-HPR, a synthetic retinoid, which is currently used in ovarian cancer chemoprevention trials in US and Italy, in animal model in vivo. Both cactus and 4HPR inhibited ovarian cancer growth. The anticarcinogenic properties of natural and synthetic retinoids have been suggested to be due, in part, to the antioxidant effect. Increased consumption of fruit and vegetables is associated with the prevention of various human diseases, and the oxidative damage is an important etiologic risk factor for many diseases, including cancer and heart diseases, the two with highest mortality rates in the US. Cactus pear extracts contain multiple antioxidants that can reduce oxidative damage. Carcinogenesis may be viewed as a process of progressive disorganization. This process is characterized by the accumulation of genotypic changes and corresponding tissue and cellular abnormalities including loss of proliferation and apoptosis controls. A dietary agent that can increase anti-proliferation pathways and change cell cycle in cancer cells without toxicity would be a potential agent for chemoprevention. Although the mechanism for cactus pear extracts in cancer prevention is unclear, our current study shows that cactus pear does alter the expression of certain genes related to cell growth and apoptosis. Cactus pear extracts increased annexin IV and decreased VEGF expression in animal tumors.

47

Annexin IV played a pivotal role in the early phases of apoptosis. The ongoing study showed that the cactus pear mixture extract increased cancer cell reactive oxygen species (ROS), which increased oxidative stress and mediated gene expression. ROS is the key regulating factor in apoptosis, which works through inducing receptor activation, regulating Caspase and Bcl-2 family, and mitochondrial dysfunction. Cactus pear mixture extract has potent anti-carcinogenesis effect and low toxicity. This holds promise for new treatment modalities in addition to prevention of ovarian cancer. The international collaborative effects are needed for fighting with cancer to reduce

cancer incidence and mortality rate worldwide. To develop the safe food-derived products would be ideal for long-term use in normal or high risk populations. Clinical trials with cactus pear mixture extract tested in high risk population are worth being developed by the international collaboration. The potential collaboration between Professor Jorge Souen and Dr. Nise Yamaguchi, Gynecologic Oncology and General Oncology group in Brazil and Dr. Molly Brewer and Dr. Changping Zou, Gynecologic Oncology group in University of Connecticut Health Center, United States will be initiated with the help from Cactus Congress, Cactus Company in US, and local universities and organizations in Brazil.

Cactus Mix Extract

e stress Oxidativ
Oxidative DNA Damage

ROS

Bax Mitochondria
Cytochrome-c Caspase-9,10 Caspase 3

Nucleus DNA Fragmentation

Cell cycle arrest

Membrane damage

Necrosis

Apoptosis

48

Antioxidant activity of cactus pear fruits and bioavailability of betalain components in healthy humans.
Maria A. Livrea
Dipartimento Farmacochimico Tossicologico e Biologico. Universit di Palermo. Facolt di Farmacia. Via M.Cipolla 74. Palermo. Italy Aerobic life is characterized by a steady formation of prooxidants, including reactive oxygen species and their products, that is approximately balanced by endogenous antioxidant defense systems. They include specific enzymes and low molecular weight molecules, much of which of dietary origin. Unfortunately, the balance is not perfect so that a certain degree of oxidative damage occurs even in healthy persons. When the generation of prooxidants greatly exceeds the cells capacity to protect itself, serious oxidative stress occurs, and the accumulation of damage will result in pathophysiological events. Consumption of fruit and vegetables is now associated with protection against various pathologies, including cancer and cardiovascular and cerebrovascular diseases, in which oxidative damage is an important etiologic factor. Various compounds from plant food, acting independently of known nutrients and micronutrients, have appeared very important in protecting human health. Most of these components can act as antioxidants or affect redox-dependent cell activities involved both in physiological and pathological events. In this scenery vegetables from the Mediterranean area, and in particular the fruits of cactus pear, have recently attracted the attention of researchers. Cactus pear (Opuntia ficus-indica) fruit contains vitamin C and characteristic phytochemicals, the redox activities of which have been reported in various scientific publications. We investigated the effects of a short-term supplementation with cactus pear fruit (500 g fruit pulp daily, for 2 wk), compared with vitamin C alone, on the total body oxidative status in healthy humans, in a randomized, crossover, double-treatment study (Tesoriere et al., 2004a). Plasma measurements, before and after the two-weeks trial, showed that consumption of cactus pear fruit positively affects the bodys redox balance, that is reduces the oxidative stress, decreases oxidative damage to lipids, and improves antioxidant status in healthy humans. Supplementation with vitamin C at a comparable dosage enhances overall antioxidant defense but does not significantly affect body oxidative stress. Among the fruit components, the characteristic pigments betanin and indicaxanthin can be considered as possible contributors to the observed in vivo effects. The radicalscavenging and antioxidant activity of these betalains has been shown in several chemical and biological models in vitro and ex vivo, including liposomes, human low density lipoproteins, and red blood cells (RBC). In addition, in experiments with cultured cells, betalains were capable of affecting redoxdriven signal transduction pathways, involved in the inflammatory response. Absorption and bioavailability are major points to characterize dietary components, and to eventually suggest sites of action in the body. Betalains are absorbed and are bioavailable from various food sources. Our kinetic studies with cactus pear fruits, in healthy humans, showed peak plasma concentrations of the micromolar order at 3 h, after a single ingestion of 500 g cactus pear fruit pulp, which provided 28 and 16 mg indicaxanthin and betanin, respectively. The urinary disposal of indicaxanthin and betanin over 12 h represented 76 % and 3.7 %, respectively, of the ingested compounds, indicating that indicaxanthin is 20-times more bioavailable than betanin (Tesoriere et al., 2004b). When compared with bioavailability of betanin from red beet juice (Kanner et al., 2001), our results show that cactus pear fruit is a good source of bioavailable betalains. Knowledge of the body distribution may aid the investigation of the eventual bioactivity of absorbed dietary components. In our study we also researched the post-absorptive distribution of indicaxanthin and betanin in circulating LDL and in RBC, and evaluated the susceptibility of the particles and cells isolated at time intervals ranging from the time of ingestion of fruit to the time of betalain disposal, to the ex vivoinduced oxidation. Both LDL and RBC incorporated betanin and indicaxanthin time-dependently, in parallel with their plasma concentration, and exhibited

49

enhanced resistance to an ex-vivo-induced oxidation, the higher the amount of betalains incorporated the higher the resistance (Tesoriere et al., 2004b; Tesoriere et al., 2005). On the overall, while suggesting major benefits from diets including cactus pear fruits, to reduce the risk of age-related and degenerative diseases in which destruction of finely tuned redox-regulated transduction pathways may play a pathogenic role, our findings open to new studies directed to possible applications of purified fruit components either as such, or as lead compounds for novel therapeutics.
Tesoriere, L.; Butera, D.; Pintaudi, A. M.; Allegra, M.; Livrea, M. A., 2004a. Supplementation with cactus pear (Opuntia ficus-indica) fruit decreases oxidative stress in healthy humans: a comparative study with vitamin C13. American Journal Clinical Nutrition, 80, 391395. Tesoriere, L.; Allegra, M.; Butera, D.; Livrea, M. A., 2004b. Absorption, excretion and distribution of dietary antioxidant betalains

in LDLs: potential health effects of betalains in humans. American Journal Clinical Nutrition 80, 941-945. Butera, D.; Tesoriere, L.; Di Gaudio, F.; Bongiorno, A.; Allegra, M.; Pintaudi, A. M.; Kohen, R.; Livrea, M. A., 2002, Antioxidant activities of Sicilian prickly pear (Opuntia ficus-indica) fruit extracts and reducing properties of its betalains: betanin and indicaxanthin. Journal of Agricultural and Food Chemistry 50, 6895-6901. Kanner, J.; Harel, S.; Granit, R. 2001, Betalains-A New Class of Dietary Cationized Antioxidants Journal of Agricultural and Food Chemistry 49, 5178-85. Tesoriere, L.; Butera, D.; Allegra, M.; Fazzari, M.; Livrea, M. A., 2005, Distribution of betalain pigments in red blood cells after consumption of cactus pear fruits and increased resistance of the cells to ex vivo induced oxidative hemolysis in humans. Journal of Agricultural and Food Chemistry 53, 1266-1270.

For a review refer to: Maria A. Livrea and Luisa Tesoriere, Health Benefits and Bioactive Components of the Fruits from Opuntia ficusindica [L.] Mill. J. PACD, 2006, pag 73-90

FRUITS OF MAIN CULTIVARS OF CACTUS PEAR FROM SICILY Opuntia ficus indica, L Mill

sanguigna

RED FRUIT

surfarina

YELLOW FRUIT

muscaredda WHITE FRUIT

8 % OF PRODUCTION
CH2OH O H O H OH H OH H H OH HO

90% OF PRODUCTION

2% OF PRODUCTION

+ N

COOH

+ N

COOH

Betanin is mainly represented in the red fruit where it is 66% of total betalains Indicaxathin is 89% of betalains of the yellow fruit, which exhibits the highest amount of betalain pigments.

The white fruit contains almost exclusively indicaxanthin

HOOC N H COOH HOOC N H COOH

Betanin

Indicaxanthin

.... and betalain pigments

50

An Overview On Medicinal And Cosmetic Uses Of Cactus In Morocco

Abderrahman Ait Hamou
ANADEC : Association Nationale de Dveloppement du Cactus. Kelaa Srghna Morocco. Email: aithamou@hotmail.com

Introduction
Cactus known as prickly pear is a widely represented crop in Moroccan landscape. In the lack of precise assessment of cactus national superficies, non-official but objective reckoning talks about at least 120.000 ha (El Yamani, 2004). In Morocco, cactus is planted above all along roadsides and as fences, but there are many regions were large orchards ensure alternative income for rural population particularly during frequent dry seasons. Pears as fruit, and pads as forage, are the main uses of cactus. During summer, the harvest season, thousands of street vendors are shown in different Moroccan cities. Cactus pear is generally consumed fresh, however, in the south, it may be sun dried and stored for later use.

cosmetic uses of cactus in Morocco.

1. Methodology
The sources of information presented in this paper are: Bibliographical data: we selected documents which are related to Moroccan context Our own field survey with: researchers, herbalists, rural women, and a sample of 25 cactus street vendors and 31 consumers. After collecting information we brushed aside declarations given by not enough interviewees.

2. Results
2.1. Documented data
Cactus was introduced into morocco by the XVIth century; so most documents referring to this specie are relatively recent in comparison with many other medicinal plants, as Aloe vera for instance, which, by the way, is called Assabr in Arabic, a word similar to Assobar, the cactus Arabic name. This precision is important because Assobar is used sometimes to name the two species. Consequently, some authors make confusion between virtues of these succulent plants. Many traditional medicine books include, more or less detailed, chapters devoted to cactus virtues. In general, the subject is treated by presenting cactus as a semi-arid region crop coming from Mexico. After pointing out main chemical characteristics of fruit, pads and flowers, treatable diseases/disorders are listed and recommended posology is described Thereby, the well-known Moroccan ethno pharmacologist Bellakhdar (1997) reported from different sources that cactus flower contains many interesting chemical components: carbohydrates, mucilage, potassium and calcium salts, and many flavonoids (luteolin, kaempferol, Quercetin,

Figure 1. Geographical distribution of cactus in morocco (Map showing high and low cactus densities areas) Pads and flowers are not consumed, but they are used for some medicinal and cosmetic purposes. In fact, there is a popular knowledge recognizing medicinal prickly pear cactuss properties. In this paper we will present a general overview of historic and current medicinal and

51

Penduletin, Rutin,..).

Cactus pads are reported to include alkaloids and bases (Hordenin, caudicin, chlorine, thyramin and mescaline). Sap is rich on mucilage. Cactus pear contains vitamin C (0.1%) polysaccharides, pentoses, hexoses, and carotenoid pigments (Betaxantins and betacyanins). While fruit and pads are poor on proteins, fig seeds are reported to contain a relatively high level of these nitrogenous substances (16%). Seeds are also rich in fats (17%) and amino acids. Many other authors report prickly pear cactus richness on nutritious components. We summarize, in table 1, the main available published data. According to several authors, different parts of prickly pear cactus are useful to treat or prevent many diseases.

Figure 2. Moroccan publication presenting OFI as a medicinal plant

Table 1: Main chemical components of Prickly pear cactus Main chemical characteristics Fruit Glucose, fructose, ascorbic acid, fats, tragacanth, sodium oxalat, pigment, yeast (saccharomyces) Glucose (%): pulp (35%) skin (21%) Fructose (%) pulp (29%) - Protein (%): 5.1 - (pulp) 8.3 (skin) Pads Flowers Fiber,- Vitamins: A, C Minerals; Ca , K Amino acids Ca (1,17 % DM); Mg (0,581); Na (1,482); K (2,46); Fe (0,039); P(0,0998); Proteins (8,49); Carbohydrates (73,09) ; Fats (2,79) ; Ash (10,48) Protein: 11.8 % Fatty acids (%): C16:0 (11.9); C16:1 (0.6); C18:0 (3.7); C18:1 (21.3); C18:2 (61.1); C18:3 (0.3); C20:0 (0.3); C20:1 (0.2) Authors Zahouili (1997) El-Kossori et al. (1987) Bellakhdar (1997) Boujenah (2005)

Seeds Seed oil

El-Kossori et al. (1987) Personal observation

2.1.1. Fruit
Fresh pear is used to stop diarrhoeas and intestinal colic (Boulous, 1983; Sijelmassi, 1996; Bellakhdar, 1997) astringent (Boulous, 1983) and anti-diabetic (Eddouks et al., 2004). There is no detail about how a sugar-rich fruit might be useful to prevent diabetes, but it seems that this ability is due to the pulp fiber. Some side-effects are however reported, they are related to fig seeds which might cause digestive disturbances such as constipation.

2.1.2. Flowers
Dried flower is mentioned among the Moroccan exported products. It is the part of

the plant the most used to treat/prevent many illnesses. Flower decoction is taken as diuretic and antidiarrheic (Bellakhdar, 1997). According to Boujenah (2005), flowers are used in traditional medication especially against the renal diseases, diabetes and constipation. Boulos (1983) reported that infusion of dried flower buds mixed with barley grains and maize silks are sold by some drug market as remedy for venereal illness, gonorrhoea and syphilis. El Hilali (2003) quoted a list of abdominal pains treated by using flower powder. He named dyspepsia, stomachic, stomach-aches, gastric ulcer, stomach ulcer, colic, colitis, heartburn, and bowels. Zahouili (1997), an herbalist, proposed a flower macerate to use as an anticramp. Flower
52

powder is a good remedy to urine retention

cactus fruit as it is testified by thousands of vendors overrunning streets in urban centres. In Casablanca during harvest season the vegetables and fruits central market reserve many stalls where are received trucks coming from far away with large quantities of figs. According to our survey with a sample of those vendors, a great part of their clients are greedy consumers because they believe that cactus pear is very beneficial for health. We were interested to know what kind of people are their main clients. Our results, shown in table 2, indicate that most of consumers are over 40 years old, as thought 44 % of interviewed street vendors.

Figure 3. Cactus dried flowers Diabetes is again treated or prevented with flower powder (Jouad et al., 2001; Tahraoui et al., 2006).

2.1.3. Pads (cladodes)

In general, cladodes are used in external applications for their nutrient and protective effects: Spineless pad is cut in 2 slices, internal face applied face-on the skin (Bellakhdar, 1997). This treatment is used in several cases: Sprain, swelling, haematoma, stemma, contusion, arthritis, lameness, rheumatism, anti-inflammatory, muscular inflammatory, myalgia, sciatic, eczema... Boulos (1983) has also reported cladodes usage as emollient and in cataplasm. Cladodes sap of Opuntia megacantha is taken, at a rate of a spoonful every morning to treat whooping-cough, and inflammation of the throat, oesophagus, stomach and intestines (Bellakhdar, 1997) and pancreas diseases (Zahouili, 1997)

Figure 4. Figs street vendors in Casablanca Responding to our question how often and why they eat cactus fruit?, a sample of consumers answered as summarized in table 3. It is important to note that more than 38 % of people interviewed prefer the fruit for health considerations. The logical next question was, how did they know that cactus fruit would be useful for their health?. We understand then that their sources of information are different: medias, neighbourhood, friends. In southern of morocco, figs are sun-dried and used as remedy for stomach ulcer. Table 2. Clients distribution according to age groups Age ranks More important Equal Total Under 20 20 to 40 3 (12%) Over 40

2.1.4. Cactus cosmetic proprieties

Available data on cactus uses as cosmetic are limited to general information rather than results coming from true investigations: crushed cladodes are known to be used as shampoo and recommended as a hair-loss preventive. Cactus sap and seed oil are used as a screen to prevent sun burns.

3. Results of Survey on cactus medicinal and cosmetic uses:

Our survey results are agreed with most of information reported previously and generate new knowledge on cactus proprieties.

6 (24%) 11 (44%) 5 (20%) 25 (100%)

3.1. Cactus fruit

Moroccan people are traditional consumers of

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Table 3. Reasons for consuming cactus fig Reasons Low cost Sweet taste Treat or/and Prevent Digestive troubles Diabetes Cholesterol Total N 4 15 6 5 1 31 % 12.90 48.39 19.35 16.13 03.23 100

3.2. Cactus flower

Six of ten herbalist shops we visited sell dried cactus flowers. They are mainly used in two situations: urine retention and kidney pains. In the first case whole flowers are infused in boiled water. The filtrate is cooled down and drunk. Generally the result is satisfactory in few hours. On the other hand, kidney stone pains are treated by infusion made with a powder including dried cactus flower and other medicinal plants (lavender, and hairy rupturewort). The treatment may last several days before stone fragments pass in the urine. According to many herbalists, asthma is also successfully treated by a medicinal recipe including: honey and cactus dried flower and jujube powders. In Rehamna region (centre of Morocco), it is very known that sheep farmers use cactus flowers to treat rams suffering from urine retention. This is one of the rare cases where cactus is used in traditional veterinary phytotherapy. Figure 5. Cactus dried flowers in an herbalist shop

Figure 6. Nopalitos or young pads: removing spines before use as shampoo. In Rabat, an aesthetician advisor recommends pads mucilage to treat feet fissures and skin crevasses (Khal, 2007; personnel communication) In the past, young pads (nopalitos) were used, mainly by rural people, as shampoo not only for their low cost but also for their advantageous effect on hair health.

3.3. Pads (cladodes)

Fresh cladodes are used both in medicinal and cosmetic purposes. Our rural survey results had led us to identify many cases where cactus pad are used. They are as follows: Child cough: cladode sap is taken as syrup. Constipation. Fractures (bones) Inflammations: eyes, bee and wasp stings, Pain: articulation, muscles (cataplasme)

3.4. Seeds oil

In the urban centres, women recognize the anti-wrinkles effects of the prickly pear seeds oil. But this product is considered too expensive for local use (1000 to 1400 euros a litre). As for argan, a woman cooperative manufacture some cosmetic products (soap, cream) by adding few seeds oil drops.

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Mohamed Ben Abdellah- Fes, INPAM Taounat), Medicinal (INRA - Rabat, University of Moulay Ismael Errachidia). Moroccan researchers are looking for cooperation with their counterparts of different nationalities. I used to be invited to attend seminars on aromatic and medicinal plants, cactus is often disregarded or referred to by few participants. Yet, as the plant has several medicinal properties, I propose to adopt Cactotherapy as a new word to differentiate this fantastical cactus herbal medicine.

Figure 7. Prickly pear seeds oil: a worthy product

4. Conclusions and Future Perspectives

This overview shows that there is a large traditional knowledge about cactus medicinal and cosmetic proprieties. A wide field is then open to research and development. This need is becoming ever more impressing because Morocco has to cope with a natural challenge (frequent drought) and cactus is viewed as one of the tools that can help to fight desertification. Thousands of hectares are yearly planted with cactus therefore a medicinal and cosmetic technology will generate additional revenue for rural population. Despite their high medicinal value due notably to fiber, cladodes are not part of food-habits of Moroccans. But in last years, there are some attempts to introduce this food to people by various ways: - Cactus industrial processing: Pickled cladodes manufactured in Tiznit (south of morocco) by Aknary women cooperative. Cladode powder: cactus is dried naturally by sun (Agadir) or advanced technology (Safi) of TV information programs, about cactus

5. References
Bellakhdar, J., 1997. La pharmacope marocaine traditionnelle: Mdecine arabe ancienne et savoirs populaires. Ibis Press, 764 p Boujenah, 2005. La Composition et valorisation des fleurs du cactus ( DR.Boujnah, INRA) Meeting on cactus Kelaa - Morocco Boulos, L., 1983. Medicinal plants of North Africa. Reference Publications, Inc., 218 St. Clair River Drive, Box 344, Algonac, Michigan 48001, 286 p., El-Hilaly Jaouad, M. Hmammouchi and Badia Lyoussi, 2003. Ethnobotanical studies and economic evaluation of medicinal plants in Taounate province (Northern Morocco). Journal of Ethnopharmacology, Volume 86, pp. 149-158 El Yamani, 2004: Troisime Journe Nationale sur le Cactus Ben Guerir- Maroc archives ANADEC Jouad, H., M. Haloui, H.Rhiouani, J.ElHilaly & M. Eddouks, 2001. Ethnobotanical survey of medicinal plants used for the treatment of diabetes, cardiac and renal diseases in the North centre of Morocco (Fez-Boulemane). Journal of Etnopharmacology, 77, 175 182. Sijelmassi, A., 1996. Les plantes mdicinales du Maroc. 4me dition. Edition le Fennec. Casablanca, Maroc. Tahraoui, A., J., El-Hilaly, Z.H. Israili, B. Lyoussi (2006): Ethnopharmacological survey of plants used in the traditional treatment of hypertension and diabetes in south-eastern Moroco (Errachidia province). Journal of Etnopharmacology. Zahouili .M (1997), (in arabic) Phytothrapie naturelle au Maroc Dar Annachr Casablanca Maroc pp.232

- Diffusion proprieties: Medias: internet

newspapers,

Meetings and publications (books, films...)

In November 2005, Moroccan cactus research was honoured. During a national event, Dra Nazareno have been in Morocco. She presented some results of her work on cactus pear as a functional food. At the same day a National Researchers Net was founded. Currently, many researchers are carrying out different investigations: Cosmetic (Univesity of

6. Useful links in Morocco

Ministry of Agriculture (Ministre de lAgriculture : www.madrep.gov.ma) Faculties and Institutes:

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INRA : Institut National de la Recherche Agronomique : www.inra.org.ma INPAM : Institut National des Plantes Aromatiques et Mdicinales : www.inpma.ac.ma CNRST : Centre National pour la Recherche Scientifique et Technique : www.cnr.ac.ma Facult des Sciences et Techniques de Settat : www.uh1.ac.ma/etablissement/fsts/

Facult des Sciences et Techniques d'Errachidia : http://www.fste.ac.ma/ Facult des Sciences et Techniques Fes : www.fst-usmba.ac.ma Non Goveremental Organistations : Aknary Women Cooperative: Tiznit email : coopaknari@yahoo.fr ANADEC: Association Nationale de Cactus: www.anadec.africa-web.org Portail de lagriculture : http://moroccontact.com/

Figure 8a. Pickled pads

Figure 8b. Cactus industrial processing (SAFI Morocco)

Figure 8c. Culinary book: a way to improve cactus consumption

56

Inhibition Of Iron Corrosion Using Opuntia Ficus Indica Extract

Hind Hammouch1, Laila Bennghmouch3, Abdellah Srhiri1, Najat Hajjaji2.
1 2

Electrochemical laboratory, Faculty of Sciences, Kenitra. Organic reactivity laboratory, Faculty of Sciences, Kenitra 3 Biochemical laboratory, Faculty of Medicine, Rabat Email : hamhind@yahoo.fr

Abstract
Opuntia ficus indica (prickly pear cactus) is a highly productive plant?and it is cultivated as a food source for both human and animals. Also cactus has important ecological roles in combating land degradation, and it is adopted and integrated in the agricultural production system of arid and semi arid zones. This work aims at developing an environmentally friendly corrosion inhibitors where the aqueous extract of Opuntia stem is used. The inhibitive action of the Opuntia extract, toward corrosion of iron in 3% sodium chloride is tested using polarisation technique and electrochemical impedance measurement. The extract examined performed important inhibitor efficiency (82-86%). Keywords: iron, corrosion inhibition, Opuntia extract

action of a natural extract from Opuntia stems was studied toward corrosion of iron in 3% chloride sodium solution, using polarisation technique and electrochemical impedance measurement The Opuntia extract contains mainly polysaccharides, mucilage which contains residues of D-galactose, D-xylose, Larabinose, L-rhamnose and D-galacturonic acid (Mc Garvie, 1979; Trachtenberg and Mayer, 1981). This composition leads us to test them to protect metals in corrosion media.

Materials and method

Stems of Opuntia were washed thoroughly, and divided into small pieces, and then it was grinded. The extract presents a high viscosity, it was diluted with distilled water, in order to facilitate the filtration of the extract and obtain a clear solution, which was used directly in the experiments. The iron studied had an elemental composition (percentage composition by weight: 0.11% C, 0.22% Si, 0.45% Mn, 0.046% Cr, 0.054%Ni, 0.01% Mo, 0.015% V, 0.0005% Nb, 0.014% Ti, 0.0003% Al, 0.002% Pb, 0.056% Cu.). The iron working electrode had a cross section of 0.78 cm2, the surface was polished, washed thoroughly with distilled water and dried with hot air. The silver electrode (Ag-AgCl) was used as the reference one, and counter electrode was of platinum. The electrolyte was an aqueous solution of 3% NaCl prepared by dissolving a NaCl PROLABO in bidistilled water. All experiments were carried out in open-air at room temperature. Electrochemical measurements were carried out after immersion of the working electrode in 3% NaCl for 30 min at the open circuit potential (Ecorr). The potentiodynamic curves were carried out using EG&G M352 Potentiostat, and impedance measurement were taken with

Introduction
Corrosion is defined as the destruction or deterioration of metals by chemical means in environments to which they are exposed (Fontana, 1987). Corrosion causes enormous damages and economic losses. Corrosion inhibitors reduce or prevent these reactions, they are adsorbed on the metal surface, and act by forming a barrier to oxygen diffusion and reducing the reaction with moisture. This inhibition effect may take place with adsorption phenomenon on metal surface and ion complex formation. Some of the inhibitors facilitate the formation of a passivating film on the metal surface (Quraishi et al., 2002). Many of the available corrosion inhibitors are highly toxic, pollute environment, and some of them are suspected as cancer causing agents. All these facts call for the research on cheap and environmentally friendly corrosion inhibitors. There are only few trials investigating the use of natural products as corrosion inhibitors (Awad, 1985; Bouyanzer and Hammouti, 2004). In this respect the corrosion inhibition

57

EG&G M6310 Potentiostat, in the frequency domain from 65 KHz to 10 MHz. The percentage of Inhibition efficiency against corrosion (E%) was calculated using the following equation (Shriri et al., 1992-1995): E% = (1- i corrin / i corrun) 100 Where icorrun is the corrosion current density without the inhibitor, and icorrin is the corrosion current density in the presence of the Opuntia extract. icorrun was determined by Tafel plot extrapolation to Ecorr, correcting the diffusion current (i) according to the relation (Shriri et al., 1992-1995): 1 / i = 1 / i* + 1 / iL [11-12] where iL is the measured limiting diffusion current, i is the measured current and i* is the corrected current.

Results and discussion Potentiodynamic Measurements

Figure 1 represents (a) anodic and (b) cathodic polarization curves of iron in 3% NaCl solution containing different concentration of Opuntia extract. The curves were plotted after immersion for 30 min in solution at the corrosion potential and under constant agitation. Corrosion parameters such as corrosion current density (Icorr), corrosion potential (Ecorr) and E % are given in Table 1. The Icorr value decreases significantly in the presence of the Opuntia extract, indicating that the extract is an effective corrosion inhibitor. The E% increases as the extract concentration increases: 59% (8%), 82% (16%).

0,1

1E-3
0,01

1E-4

I(A/Cm2)

I(A/Cm2)

1E-3

1E-5

1E-6

(0% inh) (6% inh) (8% inh) (12% inh) (16% inh)
-0,75 -0,70 -0,65 -0,60 -0,55 -0,50 -0,45

1E-4

1E-5

(0%inh) (6%inh) (8%inh) (12%inh) (16%inh)

1E-7 -0,80

1E-6 -0,60 -0,55 -0,50 -0,45 -0,40 -0,35 -0,30 -0,25 -0,20 -0,15

E(V/Ag/AgCl)

E(V/Ag/AgCl)

Figure 1. Polarization curves (a/left) anodic, (b/right) cathodic, of iron in 3% NaCl containing different concentration of Opuntia extract.

Table 1. Electrochemical parameters of iron corrosion in free and inhibited 3% NaCl solutions -Ecorr mV /Ag-AgCl 566 507 538 509 551 Icorr A/Cm2 176 72.4 49 40.7 31.4 -bc mV/dcade 42 76 79 79 70 59 72 77 82 E %

NaCl % 3 6 8 12 16

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Electrochemical Impedance Measurements

Impedance diagrams obtained for the frequency range 10 mHz-65 KHz at Ecorr for iron in 3% NaCl, are shown in figure 2. Figure 2 (a) represents impedance diagram shape obtained for the iron in 3% sodium chloride interface in the absence of the inhibitor. Two capacitive loops appear. In the context of detailed study published elsewhere (Bonnel et al., 1983), the high frequency (HF) capacitive loop is attributed to a charge transfer process, while that observed at low frequency (LF) is interpreted in terms of the mass transport which is occurring at the same time in the liquid and solid phase through the layer of corrosion products formed (Srhiri, 1985). The impedance diagrams shown in figure 2 (b), (c), (d) and (e) are those obtained for the iron in 3% NaCl solution containing different concentration of Opuntia extract. The intersection with the real axis of the extrapolated part at zero frequency gives the value (Rp +RE), where Rp is the polarisation resistance and RE is the resistance of the electrolyte. Values of the capacitance C are calculated using the relation C = (2 Fc R)-1(R representing the diameter of different loops) (Dabosi, 1991). Effect of added Opuntia capacitive loops appear: extract: three

concentration), in the high frequency (HF) range, the characteristic frequency fc, at the maximum of the imaginary part is very high ( 258Hz), and the low value of CHF = 8.2F, suggest that this HF loop is associated with the effect of a relative compact and thick inhibitor film. Inhibition efficiency calculated using values of Rp is about 86% for inhibitor concentration of 16%, this value is in good agreement with that determined from potentiodynamic measurements. Intermediate frequency loop and low frequency loop were attributed to faradic process (charge transfer process, mass transport) through a film default. In order to visual the effect of the Opuntia extract on iron electrode surface, figure 3 represents photographies of iron surface (2 cm2) in 3% sodium chloride containing different concentrations of inhibitor after 6 hours of immersion time. Photography 1 represents a reference surface of iron, before immersion in 3% sodium chloride. Photography 2 represents iron surface electrode after 6 hours of immersion in 3% sodium chloride without inhibitor, deterioration and damage on iron surface is notable caused by a corrosive medium. Photography 3 represents the iron surface in 3% NaCl containing 6% of Opuntia extract. Addition of inhibitor decreases damage on iron surface. The addition of Opuntia extract decreases markedly surface deterioration in comparison with a iron surface damaged in absence of the inhibitor. Deterioration decrease is also observed as the extract concentration is increased (photos 4 and 5)

High frequency ( HF) loop Intermediate frequency (MF) loop Low frequency (LF) loop.

For example figure 2 (d), (12% of inhibitor 1 2 3 4 5

Reference surface

Iron surface in 3% NaCl after 6 h of immersion

3% NaCl containing 6% of inhibitor after 6 h of immersion

3% NaCl containing 12% of inhibitor after 6 h of immersion

3% NaCl containing 16% of inhibitor after 6h of immersion

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Impedance diagram

100 90 80 70

NaCl 3%

-Im(ohm cm )

60 50 40 30 20 10 0 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150

2

1 Hz

0.1Hz 0.01Hz

Re(ohm cm )

Figure 2 (a)
200 180 160 140

Figure 2 (b)
8% inh

-Im(ohm cm )

120 100 80 60 40 20 0 0 20 40 60 80 100 120 140 160 180 200 220 240

0.01Hz 0.1Hz 1Hz

Re(ohm cm2)

Figure 2 (c )
240 220 200 180

Figure 2 (d)

16% inh

-Im(ohm cm )

160 140 120 100 80 60 40 20 0 0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360

0.01Hz 1Hz 10Hz 0.1Hz

Re(ohm cm )

Figure 2: Impedance diagrams of iron in free and inhibited (Opuntia extract) 3% NaCl solution

Figure 2 (e)
McGraw-Hill International, New York, p.4 Quraishi, M.A., Bhardwaj.V, and Rawat. J.; 2002. JAOCS, 79,(6) : 603-609 Awad, G.H; 1985. Proceedings of the 6th European Symposium on Corrosion Inhibitors Ann.Univ.Acier douxrara , N.S.sez V, suppl.N.8, 385-395 Ranscher, A., Kutsan, G., Bandula, R. and Szailer, T.; 2000. Proceedings of the 9th European Symposium on Corrosion Inhibitors Ann. Univ. Acier douxrara , N.S.sez.V suppl N 11 , 105-113 Greche, H.; 1999. Thse de Doctorat de 3 me cycle, Facult des sciences Knitra El.Etre, A.Y.; 2003. Corrosion Science 45, 24852495 Bouyanzer A. and Hammouti B.; 2004. Third National meeting on Electrochemistry,

Conclusion
The Opuntia extract is a good inhibitor for corrosion of iron in 3% chloride sodium solution. The results show an important inhibition efficiency about 86%. The inhibitive action increases with increasing extract concentration. This action is probably performed via adsorption of the extracts compounds on iron surface.

References
Fontana, M.G., 1987. Corrosion Engineering,

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Abstract collection:31 Mc Garvie, D. et Parolis, H. ; 1979. Carbohydrate Research 69(1) : 171-179. Mc Garvie, D. et Parolis, H. ; 1981. Carbohydrate Research 88(2) : 305-314. Trachtenberg, S. et Mayer , A. ; 1981. Phytochemistry, 20(12) :2665-2668 . Srhiri A., Derbali, Y. and Picaud, T.; 1995. Corrosion Science, 51(10) :788-796

Srhiri, A. Etman, M. and Dabosi, F.; 1992. Werkstoffe und Korrosion, 43: 406-414 Bonnel, A., Dabosi, F. Deslouis, C., Duprat, M., Keddam, M. and Tribollet, B.; 1983. J. Electrochem. Soc. 130 : 753 Srhiri, A. ; 1985. Thse de Doctorat Institut National Polytechnique de Toulouse. Dabosi, F. et al.; 1991. Journal of Applied Electrochemistry 21: 255-260.

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Network News
The most salient event of the year was the organization of the The VI International Congress on Cactus Pear and Cochineal and the VI General Meeting of the International Committee of the International Technical Cooperation Network on Cactus (FAO-CACTUSNET) held in Joo Pessoa Brazil, from October 22nd to 26th, 2007. Five partners took part in the organization of the congress: Federao da Agricultura e Pecuria da Paraba (FAEPA), Universidade Federal da Paraba (UFPB), Food and Agriculture Organization of the United Nations (FAO), Technical Cooperation Network on Cactus Pear (CACTUSNET), and International Society for Horticultural Science (ISHS). The event gathered 533 participants among producers, students and national and international researchers. The Congress also counted with the presence of national and international institutions of great importance in the area, such as the INSA, ICARDA and FAO. The Scientific Program, under the congress theme Production of Cactus Pear and other cactus in the different ecosystems in the world: challenges and technological progress, was presented in 10 Sessions, with 34 lectures and more than 230 papers presented as poster. All abstracts were published in a cd-rom that was distributed to all participants of the congress.The program was enriched by the participation of invited speakers of great importance and large experience in the area. A field visit to the cities of Juazeirinho and Tapero happened on the last day of the event (October, 26th). More than 200 people participated to the visit and were able to see by themselves all the potential, advantages and threats of the cactus pear crop. The Exhibition area was also very important for the event. It was an opportunity for 36 exhibitors (companies, institutions, researchers, artisans) from different places of Paraba and region, to show their work and products, always valuing the state and countries culture. We believe that the objectives of the Congress were reached with success. The access to the most recent advances in the area and the exchange of scientific results were promoted, many partnerships were made, as well as future projects to the development of the culture of Cactus in Brazil. The VI General Meeting of the FAO-CACTUSNET was the occasion to discuss the venue of the VIIth International Cactus Congress, and to review the composition of the International Committee of the network. Three countries were candidates to host the next congress, namely Italy, India, and Morocco. After a long discussion, it was decided that the VII international Congress on Cactus and the VII General Meeting of Cactus network will be held in India. The review of the international committee of the network has led to the following modifications: (i) deletion of the working group on socioeconomics and the introduction of a new group dealing with medicinal uses of cactus that will be leaded by Dra Monica Nazareno from the University of Santiago del Estero, Argentina; (ii) the replacement of the former coordinator of the working group on pitayas and pitahays by Dr. Noemi Judith Tel-Zur from Ben Gurion University of the Negev, Israel. The other important decisions of the network are related to the reinforcement of network activities and to the need to formalize countries focal points in order to rely more on them for network activities. Participants welcomed and recommended for both FAO and ICARDA to join their efforts to improve the network efficiency and to ensure its sustainability. Thus, in the future the network will be FAO-ICARDA Cactusnet. On the other hand intitiative will be taken by FAO and the network to identify and formalize countries focal points. Research and development institutions will be approached to have countries suggestions.

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Group picture of participants to the VIth International Congress on Cactus, Joo Pessoa Brazil, October 22nd to 26th, 2007

63

Welcome to Monica and Noemi

Dr. Monica Nazareno, Coordinator of CACTUSNET Working Group on Medicinal uses of cactus pear
Born in La Rioja, Argentina, August 22nd, 1968. Graduated from the National University of Cordoba, Crdoba, Argentina, as Chemical Licentiate (1990), she obtained her Ph. D. in Chemical Sciences in 1995 at the same institution. From 1997 she has been working as Assistant Professor at the National University of Santiago del Estero teaching Organic Chemistry to Chemistry and Agronomy students. She is currently Director of the Chemical Sciences Institute of the Faculty of Agronomy and Agroindustries. She has done her postdoctoral activities at the University of Liverpool, United Kingdom. Monica is Researcher at the National Research Council of Argentina (CONICET). She had several research stays at UNICAMP (Campinas, Brazil), University of Chile (Santiago, Chile), Poly-technique University of Madrid (Madrid, Spain). She is author of several scientific papers related to antioxidant activity and communications to national and international congresses. She coordinates national research projects. She has supervised several graduated and undergraduated thesis related to natural bioactive substances. Her main research topics are related to natural pigments, antioxidant substances and free radical mediated reactions. Institute of Chemical Sciences. Faculty of Agronomy and Agroindustries. Nacional University of Santiago del Estero. Av. Belgrano (S) 1912. Santiago del Estero. Argentina. Tel: 0385-4509528. nazareno@unse.edu.ar; manazar2004@yahoo.com.

Welcome to Dr Noemi Judith Tel-Zur, New coordinator of Pitayas and Pitahayas Working Group
Born on 16 February 1965 in Buenos Aires, Argentina, Dr Noemi Tel-Zur immigrated to Israel at the age of 20. She graduated in 1988 from The Hebrew University of Jerusalem, faculty of Agriculture and completed her M.Sc. at the same university in 1992. In 2001, Noemi obtained her Ph.D. from Ben-Gurion University of the Negev (BGU), Department of Life Sciences on Genetic relationships between vine-cacti of the genera Hylocereus and Selenicereus. Her first post-doc fellowship was supported by UCLA-BGU Program of Academic Cooperation. Her second post-doc period was spent at the Department of Horticulture, University of Wisconsin, Madison. Currently, Noemi is a lecturer at The Albert Katz Department of Dryland Biotechnologies, The Blaustein Institutes for Desert Research, Sede Boqer Campus, BGU. Her research areas are Plant Cytogenetics and Plant Reproduction with special focus in exotic fruit crops. Noemi is the recipient of and number of awards: the Pasternak Prize for Agricultural Research (2006), award for Ph.D. (Future Leadership Program for Fruit Crops in Israel, 1998-2000), and Molcho Center Scholarships (1997-1998). She is author of many publications in prestigious international journals.

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New subscribers CACTUSNET NEWSLETTER

FAO-ICARDA INTERNATIONAL TECHNICAL COOPERATION NETWORK ON CACTUS

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CACTUSNET NEWSLETTER C/O Ali Nefzaoui, General Coordinator International Center for Agricultural Research in the Dry Areas (ICARDA) Rue des Oliviers, 1. El Menzah V. 2037 Tunis, Tunisia Phone: (++216)71752134/71752099 Fax : (++216)71753170 Cellular : (++216)22347149 Email: ali.nefzaoui@icarda.org.tn; a.nefzaoui@cgiar.org

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Please include brief description of your recent current research related to cactus. We also welcome comments, stories, abstracts of your work, job or meeting announcements, requests, etc.

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INTERNATIONAL COMMITTEE OF FAO-ICARDA INTERNATIONAL TECHNICAL COOPERATION NETWORK ON CACTUS (FAO-ICARDA CACTUSNET) General Coordinator
Dr. Ali Nefzaoui, ICARDA ali.nefzaoui@icarda.org.tn; a.nefzaoui@cgiar.org

Technical Secretary
Dr. Enrique Arias, FAO Enrique.arias@fao.org

Regional Coordinators
North America Dr. Jesus Fuentes, Mexico jesus_fuentes@hotmail.com Mediterranean Basin Dr. Enza Chessa, Italy Chessa_i@uniss.it West Asia Dr. Gurbachan Singh, India gsingh@satyam.net.in South America Dr. Judith Ochoa, Argentina judith@unse.edu.ar Sub-Saharan Africa Dr. Johan Potgieter, South Africa jpot@mweb.co.za, potgieterjp@agricho.norprov.gov.za

Working Groups Coordinators

Plant Genetic resources collection, conservation & evaluation Dr. Candelario Mondragon-Jacobo, Mexico jacobo77@hotmail.com and rebe27_10@yahoo.com Fruit production Dr. Giovanni Nieddu, Italy gnieddu@uniss.it Agro-industries & post-harvest Dra. Carmen Saenz , Chile. csaenz@abello.dic.uchile.cl Forage & rangeland Mr. D. Cordeiro dos Santos, Brazil. djalma@ipa.br Nopalitos Dr. Ricardo Valdez, Mexico. vacrida@hotmail.com Cochineal Dr. Liberato Portillo, Mexico lportill@MAIZ.CUCBA.UDG.MX

Pests & diseases Dr. Jaime Mena, Mexico jmena@inifapzac.sagarpa.gob.mx Pitayas and Pitahayas Dr. Noemi Judith Tel-Zur, Israel telzur@bgu.ac.il Biology Dr. Francisco Campos, Brazil. bioplant@ufc.br Medicinal uses Dra. Monica Nazareno, Argentina. manazar2004@yahoo.com

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