**5. Laws related to collection, harvest, processing and trade in medicinal Flora**

As discussed earlier, with the growing interest in medicinal plants and ayurvedic drugs, there is a need for a long term strategy and planning to conserve and sustainable use of these plants [22–26]. Many medicinal plants like other natural resources are facing extinction and their degradation has accelerated over the years due to many reasons. Medicinal plants also face habitat destruction due to over exploitation. Though **Forest Conservation Act, 1980** and the **Wildlife Protection Act, 1972** provide some protection to medicinal plants. But as there are many medicinal floras that grow away from the domain of protected areas and there is no wellplanned strategy for their conservation, many of such unchecked floras face endangerment and become extinct. Even the floras within protected areas face depletion without proper conservation strategy. Beside this, indiscriminate and unregulated exploitation of medicinal plants for their roots, stem, bark, fruit, leaves, or whole plant leads to destructive harvesting which include 70% of the wild flora out of 95%t used [16]. If not carefully monitored, this practice could lead to the depletion of genetic stocks and ultimately to the diversity of medicinal flora.

Though, government is making efforts to manage and regulate the collection of wild flora, but a long term strategy is needed. The efforts are scattered and do not yield satisfactory results. These efforts along with the well-planned strategies and policies are needed with major involvement of local communities and indigenous habitants.

Both in-situ and ex-situ conservation strategies need to be undertaken. **In-situ conservation** involves protecting bioreserves and biodiversity hotspots, e.g. The Himalayas, Sunderbans, Eastern and Western Ghats. Such biodiversity hotspots are rich in medicinal floras, thus protection of the indigenous plant species along with collection of raw drugs need to be regulated for sustainable development of natural resources. **Ex-situ conservation** involves setting up of gene banks, herbal botanical gardens, seed banks, drug repositories, nurseries, etc. so that the endangered species can be raised again. The **Department of Biotechnology, Government of India** is working in this line and has taken various initiatives to establish gene banks.

In this regard, loss of traditional knowledge is always felt. Indigenous communities have a culture of worshipping sacred groves which is rich in plant diversity.

They know the ideal growth conditions and micro niches needed for these species to thrive. Even, tribal communities cultivate many medicinal plants for their personal use. For the large scale cultivation of such plants, there is a lot to learn from these communities. Thus, it becomes extremely important to involve tribal people, indigenous habitants in conservation programmes and to document their ethnobotanical knowledge to keep this living tradition alive.

In India, mainly three Acts cover medicinal plant issues at present. They are the **Indian Forest Act, 1927,** the **Forest Conservation Act, 1980** and the **Wildlife Protection Act, 1972**. These acts do not cover much under their domain with respect to import, export, cultivation and trade related issues. Also, they focus on plant and animal resources as a whole but there is no separate law in our country that focuses on the conservation of medicinal flora in particular. The export and import of medicinal flora in India is governed according to **Convention on International Trade in Endangered Species of Wild Fauna and Flora, 1975** commonly known as **CITES**. Being an international law CITES does not cover a vast flora which is country specific. Beside CITES, **International Union for the Conservation of Nature and Natural Resources (IUCN)** is another international organization that works in the field of nature conservation and sustainable use of natural resources. It is involved in data gathering and analysis, research, field projects, advocacy and education. In India, the **Drugs and Cosmetics Rules, 1940** defines ASU drugs; regulate their trade, misbranding, adulteration or substitution and spurious drugs use in any Herbal or Polyherbal ASU formulation. Often, local vendors and manufacturers escape such laws to promote their brand and make profit.

Unregulated harvesting of medicinal plants always takes place outside the protected areas. All over the world, the species of medicinal floras that are banned for export are due to their endangered status in the wild and not because of the medicinal value they hold. There is a need of urgent national level policy for the consolidated effort towards the conservation of medicinal plants. Policies must be formulated keeping in mind the various user groups of medicinal plants. It should advocate insitu and ex-situ conservation both. The policy should review all existing institutions working in this domain and also build new ones. Most importantly, the policy needs to take into account the legal and regulatory mechanisms related to medicinal plants.

Several efforts are afoot towards the formulation of such policies but we have yet to see something more concrete in this direction.

### **6. Forensic implications of herbal drugs**

A simple understanding about forensics is that anything which is illegal and unregulated by the laws, directly or indirectly tends to harm individuals automatically comes under the purview of Forensic Science.

With the growing trend in medical tourism, most of the western countries have travelers that travel in Asian and East Asian countries for the purpose of taking natural treatment and remedies. Such wellness centres offers a variety of herbal medications for the sale. Patients visiting such centres wrongly believe that they are being treated by natural therapy without getting any harmful chemicals and drugs administered in their body. But how much regulated such centres and their medications are, pose a serious concern. There have been a number of studies that show the adulteration and the substitution in herbal drugs as it is not much regulated by the Pharmacopeia as compared to the allopathic medicines that have strict chemical compositions and are registered under the **Drug and Cosmetic Acts** of different countries [27, 28].

The second most alarming concern about the consumption of adulterated herbal medicines is the **Herb-Drug interaction**. The contaminated herbal

**185**

*Herbal Drugs Forensic*

*DOI: http://dx.doi.org/10.5772/intechopen.98253*

government regulating agency [29].

long period of time [30].

of usage and long-term medication' [29].

**7. What can be done: a way forward**

products in developing countries [16].

like farmers and cultivators.

ble land that ensures regular supply.

preparations can have anything ranging from heavy metals to pesticides in them that may show antagonistic effects upon interacting with chemical drugs consumed by the patient. Usually patients do not share their history of consumption of herbal drugs as they think it to be natural and posing no side-effects and also there is no such prescription associated with such herbal supplements. The online websites are flooded with such nutraceuticals, dietary supplements and other herbal powder that claim relief and other health associated benefits but neither such companies nor such herbal supplements are often registered under any

Third serious issue is the addition of prescription drugs in the herbal supplements to enhance their efficacy, as this may cause serious issues in the person having allergies to certain compounds. The most common example of such adulterations is the addition of steroidal drugs. Such medications can be contraindicated

Fourthly, there is sometimes very high and potentially lethal concentrations of metals are present in these unregulated herbal drugs. When there is an unexpected death occurs, the investigating officers rigorously document all the medications, prescriptions taken by the deceased and found at the crime scene as the medico-legal experts believe the cause of death to the overdose of such prescriptions but unfortunately this is not the case with the natural herbal medicines. Also, there is near to impossible chances of detection of such medicines during autopsy as these medicines are generally consumed by the patient over the

There have been reports of Lead encephalopathy and acute lead and mercury poisoning in patients consuming herbal drugs. Roger W. Byard in his publication on 'Potential Forensic Significance of Traditional Herbal Medicines' gave examples of such toxicity in which 'a 5- year old boy who had been treated with 'Tibetan Herbal vitamins' and had ingested a total of approximately 63 g of lead over 4 years. He has given another example of a case of a 5-year-old boy with bilateral retinoblastoma whose parents relied on to a traditional remedy that caused him arsenic poisoning.' The issues quoted in the above two cases can be summarized as 'non-medically qualified healers, lack of product standards, undeclared ingredients, nondisclosure

In the light of the above stated issues, there are a number of measures that can be taken at the country as well as local level for improving the Herbal drug sector and enhancing the development of a more effective trade in medicinal plants and their

a.**Improvement of land resources:** in order to guarantee an uninterrupted supply of raw drug, there must be provisions for organized and protected cultiva-

b.**Reduction in intermediaries:** there are a number of middlemen and intermediaries involved in the market chain of herbal medicines that are needed to be reduced to avoid malfunctions and enhance the profit of primary beneficiaries

c.**Research and development:** trade and development in any country can grow with more and more investment of GDP in research and development. Research

in a person consuming non-steroidal, anti-inflammatory agents [30].

#### *Herbal Drugs Forensic DOI: http://dx.doi.org/10.5772/intechopen.98253*

preparations can have anything ranging from heavy metals to pesticides in them that may show antagonistic effects upon interacting with chemical drugs consumed by the patient. Usually patients do not share their history of consumption of herbal drugs as they think it to be natural and posing no side-effects and also there is no such prescription associated with such herbal supplements. The online websites are flooded with such nutraceuticals, dietary supplements and other herbal powder that claim relief and other health associated benefits but neither such companies nor such herbal supplements are often registered under any government regulating agency [29].

Third serious issue is the addition of prescription drugs in the herbal supplements to enhance their efficacy, as this may cause serious issues in the person having allergies to certain compounds. The most common example of such adulterations is the addition of steroidal drugs. Such medications can be contraindicated in a person consuming non-steroidal, anti-inflammatory agents [30].

Fourthly, there is sometimes very high and potentially lethal concentrations of metals are present in these unregulated herbal drugs. When there is an unexpected death occurs, the investigating officers rigorously document all the medications, prescriptions taken by the deceased and found at the crime scene as the medico-legal experts believe the cause of death to the overdose of such prescriptions but unfortunately this is not the case with the natural herbal medicines. Also, there is near to impossible chances of detection of such medicines during autopsy as these medicines are generally consumed by the patient over the long period of time [30].

There have been reports of Lead encephalopathy and acute lead and mercury poisoning in patients consuming herbal drugs. Roger W. Byard in his publication on 'Potential Forensic Significance of Traditional Herbal Medicines' gave examples of such toxicity in which 'a 5- year old boy who had been treated with 'Tibetan Herbal vitamins' and had ingested a total of approximately 63 g of lead over 4 years. He has given another example of a case of a 5-year-old boy with bilateral retinoblastoma whose parents relied on to a traditional remedy that caused him arsenic poisoning.' The issues quoted in the above two cases can be summarized as 'non-medically qualified healers, lack of product standards, undeclared ingredients, nondisclosure of usage and long-term medication' [29].


**5. Plant extract**

336 Salmonella – A Dangerous Foodborne Pathogen

*Salmonella* serotypes (James et al., 2000; Phebus et al., 1997; Whyte et al., 2003). Following the published report by Porto-Fett et al., 2010, the fermentation and drying and/or high pressure processing of contaminated dry sausage or pork are effective for inactivating *Salmonella* spp. High-pressure treatment of milk is considered to be the most promising alternative to traditional thermal treatments. Metrick et al., (1989) indicated that the pressure treatments of 310 and 379MPa/15 min at ambient temperature were required for a 3-log

The first reports on antibiotic resistant *Salmonella* had been indicated since 1960s and describe mainly case with monoresistance strain (Helmuth, 2000). In the late 1980s, the appearance multiple resistances against ampicillin, chloramphenicol, streptomycin, sulfonamides and tetracycline were found in serovar *Thyphimurium* definitive type 104 (DT 104) (Montville & Matthews, 2005). The main mechanism of bacteria exhibit resistance to antimicrobial agents can be due to many factors including drug inactivation, reduced drug accumulation, alteration of metabolic pathway and target site (Barbosa & Levy, 2000; Schwarz & Chaslus-Dancla, 2001). Much of the resistance to penicillins and cephalosporins by *Salmonella* spp. is attributable to the acquired ability of the strains to produce β-lactamase that can degrade the chemical structure of the antimicrobial agents

In recent years, the prevalence of multidrug resistant *Salmonella* in foods has been reported in many parts of the world. Several clinical treatment failures with fluoroquinolones (such as ciprofloxacin) in cases of *S. typhi* showed in Europe, Asia, and Africa (Butt et al., 2003; Nkemngu et al., 2005). Shirakawa et al. (2006) claims that the resistance to nalidixic acid and decreased susceptibility to fluoroquinolone in the *S. enterica* serovar *Typhi* isolated in Katmandu, Nepal, in 2003 were completely correlated to the mutation at codon 83 of gyrA. Most antimicrobial-resistant *Salmonella* infections are acquired from eating contaminated foods of animal origin. During 2000-2006 in Taiwan, it was found that 30.5% of the raw chicken meat was contaminated with multidrug resistant *S. enterica* serovar *Schwarzengrund* (Chen et al., 2011). Among the 88 *Salmonella* isolated from 300 meat products (raw beef, chicken meat and street foods) in Kuala Lumpur, the highest resistance was to tetracycline (73.8%), followed by sulfonamide (63.6%), streptomycin (57.9%), nalidixic acid (44.3%), trimethoprim sulfamethoxazole (19.3%), ampicillin (17.0%), chloramphenicol (10.2%) (Thong & Modarressi, 2011). The most antimicrobial resistance *S. enteritidis* isolates from South of Brazil reported by de Oliveir et al. (2005) was found in poultry related samples, where all strains were resistant to at least

The prevalence of extraintestinal *salmonella* infections caused by antibiotic resistant *Salmonella* spp. in several geographic areas of the world is increasing. Pokharel et al. (2006) demonstrated a 5% prevalence of multidrug resistance among *S. enterica* at a tertiary care hospital in Kathmandu, Nepal, with a higher rate of multidrug resistance among *S. paratyphi A* (7%) compared to *S. typhi* (3%). Rotimi et al. (2008) reported the serious problem of drug resistance in *Salmonella* spp. in Kuwait and United Arab Emirates that the non-typhoidal *Salmonella* spp. isolates from fecal samples of patients had 5-fold rise in resistant to

cefotaxime and ceftriaxone compared with reported earlier.

reduction in colony forming units (cfu) of *S. seftenberg* 775W.

**4. Antibiotic resistance** *Salmonella*

(Bush, 2003).

one antimicrobial agent.

Plant contain a variety of substance called "phytochemicals" (divided into two groups; primary and secondary metabolite), which are naturally occurring biochemicals in plants that give plants their color, flavor, smell and texture. Plant secondary metabolite differ from ubiquitous primary metabolite (e.g. carbohydrate, proteins, fats, nucleic acid) (Bako and Aguh, 2007), that have a scientifically proven effect on human health. The most important of these bioactive constituents of plants are alkaloids, tannins, flavonoids, anthraquinone, other phenolic compound and essential oil (Kisangau et al, 2007).

Extraction is the first important step for the recovery and purification of active ingredients of plant materials. Several extraction techniques and solvents are used for obtain antioxidant and antimicrobial extract from plant origin. The general techniques of medicinal plant extraction include maceration, percolation, hot continuous extraction (Soxhlet), solvent extraction, counter-current extraction, microwave-assisted extraction, ultrasound extraction (sonication), supercritical fluid extraction (Chen et al., 1992; Bicchi et al., 2000; Kaufmann and Christen, 2002). For solvent extraction method, polar solvents (such as organic acids), solvents of intermediate polarity (such as methanol, ethanol, acetone, and dichloromethane) and solvents of low polarity (such as hexane and chloroform) are used to extract plant secondary metabolites, which the extracts obtained from the same plant material with different solvent characteristics have distinct physical and biological properties. Lapornik, et al. (2005) reported that ethanol and methanol extracts of red and black currant contain twice more anthocyanins and polyphenols than water extracts, extracts made from grape marc had seven times higher values than water extracts. Among the five different Indian medical plants, methanol extract showed more antibacterial activity and moderate activity with aqueous, ethyl acetate and chloroform extract (Ashokkumar et al., 2010), while the morepolar solvent extracts (BuOH and water extracts) in Korean herbal medicines gave higher antioxidant activity than that of non-polar solvent extracts (hexane and EtOAC extracts) (Kang et al., 2003). Moreover, the chemical compound of extracts from particular plant species can vary according to the geographic origin, harvesting period and parts of the plant use. Nwokocha et al. (2011) found that all secondary metabolites analyzed were present in all tissues (leaf, stem, root and seed) studied but at different concentrations. A spatial and seasonal impact on the total phenolic content has been reported for *Poacynum henersonii*  collected at three sites in China (Hong et al., 2003).

Essential oils are a volatile liquid aromatic compound which extracted from plant cell. The cells are location in specific parts of the plant such as bark, flowers, leave, seed, peel and root. Table 1 shows the plant organ contains essential oil and their essential oil constitutes. Distillation (water distillation, water and steam distillation, steam distillation) is the most commonly used method for produces essential oils on the commercial basis (Rasooli, 2007). Some volatile oils cannot be distilled without decomposition are thus are usually obtained by expression (such as lemon oil or orange oil). The effect of diffrent distillation methods on oil content and composition of aromatic plants have been reported. The water-distillation of the rose-scented geranium (*Pelargonium* sp.) gave a higher oil yield (0.16–0.22%) than did water-steam-distillation (0.09–0.12%) or steam-distillation methods (0.06–0.18%) (Kiran et al., 2005). The oil of *Satureja rechingeri* Jamzad in full owering stage obtained by hydrodistillation, water- and steam-distillation and direct steam-distillation consisted of twenty, seventeen and twenty-two compounds, respectively, which the major constituents were carvacrol and p-cymene (Sedkon et al., 2007).

Inhibitory Effect of Plant Extracts on Salmonella spp. 339

antibacterial activity against all three *Salmonella* serotypes (*S. typhi* ATCC 19943, S*. paratyphi*

In India, Mahida & Mohan (2007) described that the methanol *Manilkara hexandra*, *Wrightia tomentora* and *Xanthium strumarium* extracts displayed MIC value of 2 mg/mL for *S. paratyphi* A whereas the methanol *Schrebera swietenoiides* and *Wrightia tomentora* showed MIC value of 4 mg/mL for *S. typhi*. The result studied by N'guessan et al. (2007) showed bactericidal effect of the aqueous extract of *Thonningia sanguinea* for all the multiple drug resistance *Salmonella* strains (*S. typhi*, *S. hadar* and *S. typhimurium)* and sensitive tested strains (*S. enteritidis*). The *S. typhimurium* strain was also found to be sensitive to extracts of *Acacia nilotica*, *Syzygium aromaticum* and *Cinnamum zeylanicum*, in Khan et al. (2009). The petroleum ether extract of *Pedalium murex* Linn exhibits the activity at 300-500 mg/disc against the *S. paratyphi A* and at 500 mg/disc against the *S. paratyphi B* (Nalini et al., 2011). Furthermore, the root of the *Euphorbia balsamifera* has high activity against the *S. typhimurium* when compared with the leaves and stems extracts (Kamba & Hassa, 2010). In contrast, the extract of eucalyptus from root, leave and stem had exhibited activity against *S.*

**7.** *Salmonella* **control in food product and food packaging by plant extract**

Nowadays, the foodborne outbreaks *Salmonella* food poisoning and the prevalence of antibiotic resistant *Salmonella* in humans, animals and food are increasing (Rabsch et al., 2001; Angulo et al., 2000; O'Brien, 2002). Consumers are also concerned about the safety of food containing synthetic preservative. Therefore, there has been growing interesting in using natural antibacterial extract from herb or spice for food conservation (Smid & Gorris, 1999; Fasseas et al., 2008; Gutierrez et al., 2008). Particular interest has been focused on the potential application of plant extract or essential oils as safer additives for meat, poultry,

The combination of the oregano essential oil at 0.6% with nisin at 500 IU/g showed stronger antimicrobial activity against *S. enteritidis* in minced sheep meat than the oregano EO at 0.6% but lower than the combination with nisin at 1000 IU/g (Govaris et al., 2010). The minimum inhibitory concentration of the Capsicum extract to prevent the growth of *S. typhimurium* in minced beef was 1.5 mL/100 g of meat; the addition of 1%, 2%, 3% and 4% w/w of sodium chloride did not have any additional inhibitory effect on *Salmonella* (Careaga et al., 2003). Ravishankar et al. (2009) suggest that the food industry and consumers could use apple-based edible films containing cinnamaldehyde or carvacrol as wrappings to control surface contamination by foodborne pathogenic microorganisms, which at 23°C on chicken breasts, films with 3% antimicrobials showed the highest reductions (4.3 to 6.8 log cfu/g) of both *S. enterica* and *E. coli* O157:H7. Moreover, the lowest concentration of trans-cinnamaldehyde (10 mM) reducing *S. enteritidis* populations inoculated on chicken cecal contents by approximately 6.0 log(10) cfu/mL after 8 h and >8.0 log(10) cfu/mL after 24 h of incubation (Johny et al., 2010). The carvacrol vapour was effective at preventing growth of *Salmonella* on agar and in significantly reducing viable numbers on raw chicken at temperatures ranging from 4°C to 37 °C (Burt et al., 2007). The results by Shan et al. (2011) showed that the five spice and herb extracts (cinnamon stick, oregano, clove, pomegranate peel, and grape seed) were effective against *S. enterica* in cheese at room temperature (~23°C), which the clove showed the

A and *S. gallinarum* ATCC 9184) (Lee et al., 2006).

*typhi* (Evans et al., 2002).

milk, fruit and vegetable.

highest antibacterial activity.

Table 1. Essential oils in each plant organ (Base on Krishnasamy, 2008)