INTRODUCTION:

Herbal diversity and its therapeutic application in traditional medicines related with their efficacies assessed generally in one-dimensional ways. Many plant species and crude drugs are applied for different diseases and symptoms within one community and varies to larger extend between different communities. Large percentage of rural population has been using herbal crude drugs or products to rejuvenate their health, either alone or in combination with other herbals. According to the WHO, 75% of the global populations are employing herbs for fundamental immediate healthcare needs¹.

The quantity of usage denotes a herbal “renaissance” around the world. It is high time to document such indigenous knowledge about herbals². Herbal medicines have advantages over synthetic formulations, because of their pool of phytochemicals with biological potentialities^3,
4. Many Ayurvedic herbals are known for their phytoconstituent⁵ and therapeutic applications such as health, promote hygiene⁶, skin diseases⁷ and to treat neurodegenerative disorders including Alzheimer's⁸. Traditional knowledge of medicine is transferred from generations to generation. The indigenous knowledge implies beliefs, claims, drug formulation, practices in alleviating disorders and disease among the local and aboriginal populations⁹.

Bryophytes become part of ethno-medicinal remedies in traditional medicine and inspired researchers to explore their potential through biotechnology and bioengineering. Currently, the species are recorded with wide range of usages ranging from lead products in medicine, agroecology, flavouring agents, biofuels, climate change bioindicators, filtering of pollutants, gardening and in floristic decorations¹⁰. Many local communities from Poland, Argentina, South, North and Eastern India, Australia, Africa, America, Europe, New Zealand, Turkey, Pakistan, China, Nepal Japan and Taiwan use many species of bryophytes as ethno-medicine. Mosses like Bryum, Hypnum and Fissidens growing in association with others act as rock builders. Bryophytes offer a high complexity of biochemical pathways, as a consequence of their environmental and genomic diversification. The pool of phytochemicals extracted from them is growing drastically. The bio wealth of this group opens avenues to functional and comparative genomics studies, including mining for gene groups and tracing the evolutionary role of metabolic cycles. Alvarez et al.,¹¹activated shikimate, phenylpropanoid, oxylipins, and auxin pathways in Pectobacterium carotovorum via elicitors-treated moss reflecting its application in medicine. Asakawa and Ludwiczuk¹²profiled the phyto molecules of bryophytes and their biological activities. Asakawa et al.,¹³ reported unique bibenzyls, prenyl bibenzyls, bis-bibenzyls, and terpenoids from the liverwort Radula. Even though, there was minimal scientific outputs among bryophytes, but still many high range areas people rely on them for the effective cure of different ailments. In this scenario, the present study was undertaken to document the ethno-medicinal usage of bryophytes from selected districts of Kerala.

MATERIALS AND METHODS:

The native herbal vendors carry their traditional drug sale at public places such as bus, railway stations, Govt. offices, temples, pilgrim areas, highways or in local markets and Sunday markets. The survey was carried in selected eight districts such as Idukki, Kozhikode, Palakkad, Wayanad, Pathanamthitta, Thiruvananthapuram, Kollam and Thrissur which were known for herbal vendors¹⁴. Silent observation of the business of the local vendors was initially recorded followed by oral interviews. Most of them were known and experienced nattu vaidhyans. Customers were local people and also from other areas of the district. The customers were also interviewed regarding their experience with the vendors. The native vendors were showed the plants, type of recipes, name of disease, method of administration, duration and doses. The plants were identified using relevant floras and authenticated with herbaria of University of Calicut.

RESULTS AND DISCUSSION:

Bryophytes are the earliest land plants originated during unfavorable environmental condition and their by inevitably synthesized a pool of phytochemicals to tide over such diverse harsh conditions. The secondary metabolites reported in the various species have remarkable biological, ecological and evolutionary importance¹⁵. No serious attempts were carried in exploring this neglected group. The molecules so far reported were vitamin B derivatives, alpha- tocopherols, unsaturated fatty acids, phenolics, enantiomeric mono, sesqui, diterpenoids, bibenzyls, bis-bibenzyls and polyketides¹⁶.

In the present study, 24 species were recorded from the vendors knowledge linked to medicinal usage for curing many human/animal ailments (Table 1). Marchantia polymorpha was employed to treat bites, boils, burns, cuts, eczema, and wounds. Jantwal et al.,¹⁷ reviewed microbicidal and anti- inflammatory potentials of Marchantia with the secondary metabolites such as flavonoid, tannins and phenolics. They also recorded the lead molecule Marchantian A as an antimetastatic agent. So-Yeon et al.,¹⁸ also proved the anti-Inflammatory efficacies of the bryophytes Marchantia polymorpha and Racomitrium canescens. Thus, the vendor knowledge recorded in the present study was justifiable and they used this practice from time immemorial periods.

Table 1. List of bryophytes and its applications to cure diseases

Si. No	Plant Name	Recipe	Disease	Period
1	Marchantia polymorpha L.	Paste + salt	Bites, boils, burns, cuts, eczema, wounds and inflammation	7-14 days
2	Rhodobryum roseum (Hedw.) Limpr.	Decoction	Nervous prostration and cardiovascular diseases, sedative	45 days
3	Polytrichum commune Hedw.	Decoction + honey	Diuretic, stones of kidney and gall bladder and hair growth	30 days
4	Fissidens nobilis Griff.	Oral intake	Diuretic, hair growth, mouth ulcers, leucorrhoea and malaria	14 days
5	Taxiphyllum taxirameum (Mitt.) M. Fleisch.	Extract/ paste	Haemostasis and external wounds	21 days
6	Mnium cuspidatum Hedw.	Extract/ paste	Hematostasis, pain of burns	14 days
7	Riccardia multifida (L.) Gray	Extract of Taxiphyllum and Riccardia species/ paste	Antileukemic and Stomachache and swellings in cattle	60 days
8	Barbula indica (Hook.) Spreng.	Extract + pepper+ betel leaf/ paste	Human epidermoid carcinoma Menstrual pain , fever and decoction	120 days
9	Hyophila attenuata Broth.	Paste of Hyophila + Bryum + Castor oil	Earache, fungal skin infections	14 days
10	Bryum argenteum Hedw.	Paste	Burns/bruises/wounds	21 days
11	Dumortiera hirsuta (Sw.) Nees	Oral intake	Infections	10 days
12	Hypnum cupressiforme Hedw.	Extract	Insecticidal	30 days
13	Plagiochila beddomei Steph.	Paste	Wound healing
14	Riccia fluitans L.	Paste	Infection	21 days
15	Pallavicinia lyellii Gray. sp	Paste	Antifungal	21 days
16	Thuidium tamariscellum (C. Muell.) Bosch. and Sande-Lac	Paste	Antibacterial	21 days
17	Brachythecium buchananii (Hook.) A. Jaeger.	Paste	Antibacterial	21 days
18	Philonotis fontana (Hedw.) Brid.	Paste	Relieve pain of burn and heal burns, adenopharyngitis, antipyretic,	21 days
19	Targionia hypophylla L.	Paste	Extract + leaves of cassia tora ground in to a paste and mixed with two tablespoons of coconut oil and smeared over the body of the children affected by scabies, itches and other skin diseases	14 days
20	Leucobryum bowringii Mitt.	Extract	Body pain, paste of leaf tips mixed in toddy	10 days
21	Funaria hygrometrica Hedw.	Oral/ Paste	Hemostasis, pulmonary tuberculosis, bruises, skin infection	21 days
22	Octoblepharum albidum Hedw.	Extract	Febrifuge and anodyne	21 days
23	Ditrichum pallidum (Hedw.) Hampe	Paste	Convulsions, particularly in infants	21 days
24	Weisia viridula (L.) Hedw.	Decoction	Cold and fever	10 days

Rhodobryum roseum was traded by the vendors for nervous prostration and cardiovascular diseases and also as sedative. These features may be due to the phytochemicals present in the species. R. giganteum and R. roseum of Bryaceae are referred as Huixincao herb in folk drug of Yunnan province, China, especially by diverse ethnic minority groups. Traditional knowledge of herbs contribute for treatment of cardiovascular diseases⁴. Li et al.,¹⁹ fractionated volatile fractions from R. giganteum by solid-phase micro extraction and gas chromatography/mass spectrometry. Polytrichum commune was traditionally used as diuretic, to dissolve stones of kidney and gall bladder and induction of hair growth. It is also used in pillow and mattress stuffing for healthy sleep. No serious scientific works are documented in the species. Greeshma et al.,²⁰ also documented the traditional knowledge (TK) of the species from Neyyar wildlife sanctuary, Trivandrum. No scientific validation is available related the usage of the species still now.

Hypnum cupressiforme was traded as effective to eradicate insect pests i.e., biopesticide. Lunic et al.,²¹ characterized phenolic acids like p-hydroxybenzoate, protocatechate p-coumarate, gallate, and caffeate and evaluated the efficacy of immune-modulatory activities of this species under in vitro condition. Many research outputs justify that phenolic acids are good insect deterrents²². Mohamad Samini²³ reported that quercetin have protective effects against toxic agents in tissues including liver, kidney, brain, lung and heart.
Dumortiera hirsuta was sold as curative against infections. Liu H-P et al.,²⁴ proved the inhibitory potential of Riccardin D from D. hirsuta against intestinal adenoma formation in APCMIN/+mice. Hence, the TK was justifiable in terms of the lead molecule recorded in the species. Taxiphyllum taxirameum was used for hemostasis and external wounds in Western and Eastern Himalayas, South and Central India was reported by Alam²⁵. Mnium cuspidatum used for curing wounds formed during burns. Pejin et al.,²⁶ reported that Mnium hornum as a promising source of arachidonic acid and its medicinal usage.

Riccardia multifida was employed as antileukemic and relief from stomach ache, swellings in cattle. Asakawa et al.,²⁷ isolated Riccardin A and riccardin B, two novel cyclic bis(bibenzyls) with cytotoxic properties from Riccardia multifida. Barbula indica was used for human epidermoid carcinoma, menstrual pain and fever. Adebiyi et al.,²⁸ screened the medicinal secondary metabolites such as alkaloids, flavonoids, phenols, saponins and steroids from the species. Hyophila attenuata was traditionally used for curing ear ache and fungal skin infections. According to Chandrasekar⁷ herbal constituents are more effective in the management of skin diseases So far no validation was recorded about this knowledge. Bryum argenteum was recommended by the vendor for burns/bruises/wounds. Sabovljevic et al.,²⁹ proved the antiseptic power of the species. Leucobryum bowringii was used against body pain. The mode of preparation was paste form mixed in toddy. Manoj et al.,³⁰ established the efficacy of methanolic and water extract of Leucobryum bowringii on MCF 7 malignant cells.

Wankhede³¹ screened Funaria hygrometrica the moss by GC-MS and reported a pool of components such as 3, 7, 11, 15-Tetramethyl-2-hexadecen-1-ol, diterpene alcohol, Heptadecane, 2, 6, 10, 15-tetramethyl fractions. Traditional usage connected with hemostasis, pulmonary tuberculosis, bruises, skin infection may be due to the phytochemicals and the usage was justifiable. Weisia viridula was used to treat cold and fever as per the vendors knowledge. Asakawa³²recorded biologically active compounds from this species. Satish Chandra et al.,³³ also recorded ethno-medicinal features of the species connected with hepatic disorders. Ditrichum pallidum was employed for convulsions, particularly in infants. No scientific validation was recorded in the literature survey. Philonotis fontana of Bartramiaceae was used to cure relieve from pain of burn and heal burns, adenopharyngitis and antipyretic. No substantiating evidence was recorded in the literature survey. The thalloid Targionia hypophylla was used by Irular tribe of the Attappady valleys of Kerala state to cure skin diseases due to the simile of the liverwort to the rough surface of the diseased part. Frullania ericoides is applied for hair-related afflictions by tribal people of South India due to long-stemmed and hair-like thallus. Targionia hypophylla were used for healing efficiency on incision and excision in in vivo wounds models (24.1% increase in tensile strength on the linear incision wound test)³⁴ . Plagiochila beddomei was used by peoples in Melghat region of India for treating skin diseases. Manoj and Murugan³⁵ proved the wound healing and angiogenesis potential of aqueous and methanolic extracts of the species. Similarly, the antioxidant and antimicrobial activity of methanol extract of the species was also reported³⁶. Shivani Sharma et al.,² reported the beneficial effects of plants having antioxidant activity and the role of herbal products in thefight against deleterious diseases. Riccia fluitans indicated no ability to inhibit growth of Pseudomonas aeruginosa, Staphylococcus aureus or yeast³⁷. Meanwhile, Pates and Madsen ³⁷documented the antimicrobial substances in chlorophyllose plants growing in Florida. Glime,³⁸reported medical uses and biologically active substances from the species. Andrea et al.,³⁹ proved the antibacterial power of the species in terms of inhibiting bacterial growth. Bhagwat and Chavre⁴⁰also documented the medicinal potentiality of bryophytes. Subramoniam and Subhisha⁴¹established the in vitro antifungal activity of P. lyellii against the fungal species such as A. niger, A. fumigatus, F. oxysporum and C. albicans using disc diffusion and direct dilution methods. Aqueous, alcoholic and hexane extracts were tested and the most active alcoholic extract was subjected to sequential solvent fractionation. Hexane fraction was promising and subjected to thin layer chromatography on silica gel and each spot on the gel was tested for activity and the active spot was chemically analyzed. The alcoholic extract was evaluated for short-term toxicity in mice. Water, alcoholic and hexane extracts of P. lyellii showed varying levels of activity against the test fungi; alcohol extract exhibited maximum activity. Out of the 4 tested fungi A. fumigatus was the most sensitive. The alcoholic extract was devoid of conspicuous short-term toxicity to mice. Active hexane fraction was separated from alcohol extract and from this a steroid component with remarkable antifungal activity was isolated using thin layer chromatography (TLC). Greeshma and Murugan⁴² validated the presence of terpenoids in Thuidium tamariscellum and its antioxidant potentialities in terms of DPPH, ABTS, H₂O₂, FRAP and metal chelating ability. Further, the FTIR analysis of chloroform, ethyl acetate, petroleum ether and methanol extract confirmed the presence of primary, secondary amines, amides, alkanes, aldehydes, saturated aliphatic alkenes, alcohols/phenols, aromatic amines, nitro compounds, esters, ethers, aliphatic amines, carboxylic acids, alkyl halides and carbonyls in the moss indicating the presence of medicinally important compounds like terpenoids, flavonoids and alkaloids in the various solvent extracts of the moss. Further, Greeshma and Murugan⁴³ extracted, purified and fractionated terpenoids and evaluated its microbicidal potentialities from Thuidium tamariscellum and Brachythecium buchananii against E. coli, S. aureus, P.aeroginosa, S. mutans and B.cereus. Fungal species included were Aspergillus flavus and Candida albicans. The analysis included susceptibility determination test, effect of the extract on cell membrane integrity and fungal spore inhibition assay. The values ranged from 0.0625 to 0.125mg/mL and 0.25 to 0.5mg/mL against S. aureus and E. coli respectively. Both the moss species showed remarkable antifungal properties.

Octoblepharum albidum (Calymperaceae) was proved to have analgesic, antipyretic, antioxidant, antibiotic, and microbicidal activities^44,45. Kishore Naidu et al.,⁴⁶isolated daucosterol and friedelin from the ethyl acetate extract of the species and proved its in vitro anti-inflammatory power by protein denaturation method and anticancer via SRB assay using ovary, cervical and lung cancer cell lines. Rachana and Verma⁴⁷reported that methanolic extracts of liverwort Sauchia spongiosa, Marcantia polymorpha, Conocephalum conium, have fungicidal activity and Fissidens sps and Polytrichum possessed diuretic potentiality. According to India’s folklore, extracts of Octoblepharum albidum was used in the treatment of bacterial infections, fever and diabetes. Tatipamula et al.,⁴⁸ profiled the ethanolic extract using column chromatography and fractionated 6-hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydrobenzo furan-2(4 H )-one( 1 ), 4,4,7a-trimethyl-2-(prop-1-en-2-yl)-2,4,5,6,7,7a-hexahydrobenzofuran-6-ol( 2) and 1-(6-hydroxy-2-(2-hydroxypropan-2-yl)-4-methoxy-2,3-dihydrobenzofuran-7-yl)-ethan-1-one( 3 ). The extract revealed ferric ions, DPPH free-radicals, superoxide free-radical scavenging potential and α-glucosidase, α-amylase and aldose-reductase efficacy in glycemic albino rats. Histopathological examination of the pancreas substantiated the assay data.

CONCLUSION:

Currently, large proportion of rural population believes in traditional knowledge for treating diseases or disorders. Globally, the usage bryophytes against skin, heart, eye and reproductive problems were documented in literature. Some of them were scientifically validated and others not yet. Unfortunately the lower primitive plant group is ignored by the researchers and scientists. These bryophytes may provide remedies to many of the life style disorders with minimal side effects. It is high time for pharmaceutical industries to pay attention toward these ignored plants for evaluation and validation of such traditional knowledge practices to produce novel drugs and would be an effective strategy for the improvement of human health care.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

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Received on 10.12.2021 Modified on 19.05.2022

Research J. Pharm. and Tech 2023; 16(4):1895-1900.

DOI: 10.52711/0974-360X.2023.00311