1. Introduction
The genus
Gaultheria, belonging to family Ericaceae, are berry producing shrubs found in many parts of the world [
1]. Few species of Gaultheria (
G. trichophylla Royle.) and others are confined to Western Himalayas in their distribution [
2]. In Uttarakand, India, alpine and sub alpine regions of Western Himalaya
G. trichophylla Royle are used as appetizers in the form of fruits [
3]. In Chinese herbal medicine,
Gaultheria yunnanensis is traditionally used to treat swelling, rheumatoid arthritis, and pain [
4]. To treat headaches,
G. trichophylla is applied to the forehead [
5]. Violet fruits of
G. trichophylla are used in small amounts daily to treat asthma in temperate Himalaya Himachal Pradesh [
6]. Winter green oil naturally obtained from
Gaultheria is used in drug and food industries to enhance the scent and flavor of toothpaste, mouthwash, and confectioners. It is also used medicinally to relieve muscle and nerve pain as a complementary medicine in aromatherapy. Industrial usage includes using it as an antibiotic and an insecticide [
7].
Describing herbal drugs in a systematic manner is based on multiple approaches of pharmacognostic, taxonomic, and chemical analysis, including documentation of their biological and geographical source, cultivation, collection, processing, morphological, microscopic, and chemical characters [
8]. The herbal industries and indigenous communities of the Indo-Pak Sub-Continent commonly face problems in proper authentication and identification of the herbal drugs. They are made misguided and deal with completely different taxa [
9]. The raw materials used by the pharmaceutical industry and people are usually obtained from the market, which may be contaminated, substituted, or adulterated accidentally or deliberately [
10]. The drug identification involves physical, chemical, biochemical, and biological features [
11]. To verify the medicinal nature of a drug, multi-solvent similarity methods of cold and hot treatment are used to verify the drug effectively [
12].
The interest in traditional Phytomedicine products is increasing day by day throughout the world. It is highly significant to know the nature and safety of such plants for consumption purposes. The plants’ samples must be screened out for safe levels of heavy and toxic elements such as Cu, As, Cd, Pb, and Hg to meet the safety threshold of the WHO. The life-essential elements such as P, K, and Na are important for keeping normal health; therefore, in order to recommend them for nutrition, quantification of these elements is highly significant [
13]. These minerals (Zn, Cu, Fe, Pb, Mn, Cr, Ni, Sr, and Co) are necessary in smaller amounts. The absence or excess of these elements may cause abnormalities to the human body [
14]. Sodium is essential for keeping the balance of inside and outside fluid and in regulating the volume of blood [
15]. Magnesium (Mg) contents are important to control circulatory diseases and improve the dietary quality of plants, animals, and humans [
16]. Calcium (Ca) is an important component for the development of bones and teeth [
17]. Other than medicinal values, the plants and their polyphenolic compounds have become the focus of current nutritional interest due to their health promoting effects [
18]. Different plant tissues such as vegetables, fruits, seeds, leaves, roots, and floral parts are enriched with naturally occurring phytochemicals known as phenolic compounds [
19]. The oxidative stress can be treated with flavonoids naturally taken as human foods like catechin, quercetin, hypersoid, myricetin, and rutin [
20].
Food poisoning is measured as one of the major general reasons for illness and death in developing countries. Different causal agents are responsible for food spoilage or food borne diseases such as
Staphylococcus aureus, Bacillus cereus, as well as members of Gram-negative bacteria like
Salmonella typhi,
Escherichia coli, and
Pseudomonas aeruginosa [
21]. Global reduction in wheat yield is caused by a serious disease (HLB) called leaf blotch disease of Helminthosporium [
22]. The soyabean yield and quality is seriously reduced due to
Fusarium solani, which causes root rot disease [
23]. Fungus such as
Aspergillus flavus causes invasive aspergillosis which is symbolized in intensive care patients by complications such as severe influenza pneumonia [
24]. A filamentous, saprophytic fungus known as
Aspergillus fumigatus causes several pulmonary ailments in birds, humans, and other mammals [
25].
Phytotherapy is the use of specific medicinal plants and their isolated compounds of bio-active nature for health care and treatment of diseases [
26]. To cure such diseases, the plant produces certain chemical compounds called secondary metabolites. Synthetic chemotherapeutic agents are associated with a number of side effects, whereas the plant secondary metabolites are comparatively safe and associated with low or no side effects. Therefore, scientists are trying to isolate antimicrobial agents from plant origin. In doing so, initially the plant extracts are tested for antimicrobial spectrum. Researchers in different parts of the world have studied and screened out the effects of plant extracts on microorganisms [
27]. Extracts obtained from plants are often used in phytochemical research. Different bioassays are used to monitor efficacy of extracts and fractions. In the screening of cytotoxic and biological activities an economical and rapid test, brine shrimp (
Artemia salina) test (BST), has been used [
28].
Several plant species have been evaluated for their antimicrobial activity in the past twenty years [
29]. Some plants tested for antimicrobial activity confirmed positive results [
30]. Therefore, it is time to develop novel antimicrobial agents particularly resistant to conventional antimicrobials [
31]. Developments of new drug sources involves exploration of different plants to be tested as therapeutic agents, as being living organisms, they also suffer from different diseases [
32]. Most of the species of
Gaultheria are edible and useful for the health of humans due to their good level of polyphenol; however, extensive studies are required for the toxicity testing, nutrients, and chemical compounds on the Gaultheria [
33]. The species of genus Gaultheria grows very slowly, therefore, they are difficult to cultivate [
34].
Due to the increasing demands of the huge population of the world and the alarming problem of the drug resistance developed by microbes, the present study is aimed to determine antimicrobial activities of the selected plant and correlate the observed biological potential with major phytochemical groups: phenolics and flavonoids.
4. Discussion
In the south west district of China,
Gaultheria yunnanensis (Franch.) Rehd. from the family Ericaceae is widely used herbal medicine to treat rheumatoid arthritis, trauma, swelling pains, chronic tracheitis, cold, and vertigo. Analgesic and antipyretic effects of this plant are due to its volatile oil called “wintergreen oil” [
47]. From the pharmacognostic evaluation it can be concluded that the powder drug of
G. trichophylla can be differentiated and authenticated on the basis of fluorescence and solubility analysis. The various solvents (C
6H
6 and HNO
3) used in the current study for pharmacognostic characterization may be recommended as a reference for future studies. Qualitative study of phytochemicals suggested the presence of various active constituents, while an appraisable quantity of phenolics and flavonoids were also reported in a quantitative estimation of different extracts. The current findings of the total phenolic and flavonoids contents also agreed with [
24]. Flavonoids estimation with TLC showed the presence of four flavonoids. The presence of phytoconstituents in
G. trichophylla such as rutin, quercetin, and gallic acid was also confirmed using HPLC techniques by [
48]. According to [
47], some salicylate derivatives, organic acids, lignans, diterpenoids, triterpenoids, sterols, coumarins, and flavonoids have been isolated and identified from the roots of
G. yunnanensis. From the study of [
4], two methyl salicylate glycosides were naturally isolated from
G. yunnanensis. These results also showed consistency with those of [
49]. Elemental analysis of the current work revealed important nutritional minerals which are parallel to the findings of [
50].
The outcomes of cytotoxic activity revealed moderate cytotoxic activity. Using extracts of
G. trichophylla [
48] and working on fruits of
Gaultheria pumila [
51] reported excellent anticancer activity against the human cancerous cell line. Conclusion of results against said fungal strains revealed that methanolic extracts of
G. trichophylla were found more efficient fallowed by ethyl acetate and n-hexane extracts. According to [
52,
53] antifungal properties are due to saponins.
Antibacterial activities conclude that methanolic, acetate, and n-hexane extracts were found more efficient against all the aforementioned bacterial strains. The same extracts of the
Toddalia asiatica leaf and stem were found active against gram positive and gram-negative bacterial strains according to [
54]; similar results were also reported by [
43]. Inhibitory effects against various pathogenic bacterial strains of plant extracts can be compared with phenolic composition [
55,
56]. Crude catechins were found more efficient against gram positive bacteria [
51].
A classification for antifungal activities based on MIC values of extracts was proposed by [
57]. Extracts considered to be strong inhibitors have MIC values of 500 ug/mL, while extracts having MIC values of 600–1500 ug/mL are considered as moderate inhibitors, and extract considered as weak inhibitors have MIC values above 1600 ug/mL. In this contest, our current study showed antifungal activity of all fractions against the said pathogen; however, our scheme was slightly modified and the fractions that inhibited growth of fungus by more than 70% were considered as strong inhibitors, those having 60–70% were considered as moderate, and below 50% were considered as weak inhibitors. The outcomes of current work defended the use of investigated plant in the alpine and sub-alpine region of Western Himalaya as ethno-medicine.