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Malaysian Lost National Treasure:
A valuable “Tiger-Milk” Medicinal Mushroom (Lignosus rhinocerotis).
*Tan CS1, Ng ST2, Yeannie Yap HY3, Lee SS3, LeeML1, Fung SY3, and Tan NH3.
1Strategy Research Centre, MARDI, P.O. Box 12301, 50774 Kuala Lumpur;
2Ligno Biotech Sdn Bhd, Tmn Perindustrian Balakong Jaya 2, 43300 Balakong Jaya;
3Department of Molecular Medicine, University Malaya, 50603 Kuala Lumpur.
*Email: cstan@mardi.gov.my, tanchonseng@gmail.com
摘要 马来西亚国宝, 虎乳芝 (Lignosus rhinocerotis,当地人称“老虎奶”,“cendawann susu rimau
(harimau)”),是著名的传统药用真菌。其药用纪录可以追溯到1664 年。虎乳芝的药用部分是菌核。它的
形成。 L. rhinocerotis 的生命周期为2~3 年。我们也开发了虎乳芝菌核冻干粉以作为药用产品的原料。我
程,L. rhinocerotis 品种的開发进展及其生物药理属性的研究,以验证其传统药用功效。科研成果已经证明
中文关键词 虎乳芝;药用真菌;哮喘;免疫调节;抗炎;癌症。
ABSTRACT Tiger Milk Mushroom (also known as “cendawan susu rimau (harimau)”, Lignosus rhinocerotis),
hailed as Malaysia’s national treasure has been used as a medicinal mushroom since 1664 by local communities to
treat asthma, fever, cough, cold, sinusitis, cancer, food poisoning, for wound healing and as a general tonic.
Despite its high medicinal values, little attempts had been made to study it, due to the very limited supply as it
can only be wild collected from the forest by chance, hence hindering the discovery of its full potential for health
and economical benefits. We have now successfully cultivated the mushroom in a controlled environment
producing high yield and the formation of sclerotia which eventually led to development of stem and formation of
the mushroom cup. This paper reports the 10 years research and development journey of the development of L.
rhinocerotis cultivar, the investigations of its safety and efficacy as part of the attempt to validate the medicinal
benefits and ethno-botanical claims of Tiger Milk Mushroom. The scientific research findings have shown that
the medicinal benefits of Tiger Milk Mushroom sclerotia are attributed to its anti-inflammatory,
immunomodulating and anti-cancer properties.
Keywords Lignosus rhinocerotis;Medicinal mushroom;Asthma;Anti-inflammatory;Immune-modulating;
Some 400 hundred years ago, an important medicinal product was given to an European government agent who
sailed to this region. The Diary of John Evelyn (publication dated 22 June 1664) recorded the name of the
medicinal product as “Lac tygridis” (meaning tiger’s milk). It was described as a fungus used by local
communities to treat diseases that European doctors could find no cure to. This fungus was also clearly
described by Sir Henry Nicholas Ridley, “the father of Malaya's rubber industry” (Ridley, 1890), as an important
medicinal mushroom used by the local communities. In the year 1890, Cooke pioneered the scientific
documentation of this fungus and named it as Fomes rhinocerotis using a specimen obtained from Penang. The
scientific synonym commonly used currently is Lignosus rhinocerus/ rhinocerotis.
The Tiger Milk mushroom (L. rhinocerotis) is an important medicinal mushroom in Southeast Asia and is one of
the most popular medicinal mushrooms used by indigenous communities of Peninsular Malaysia (Lee et al., 2009).
The Tiger Milk mushroom, “Cendawan susu rimau (harimau)” has traditionally been used by the Malays, Chinese
The 9th World Congress of Chinese Medicine
中 药 研 究
Study on traditional Chinese medicinals
and indigenous communities in Malaysia for treatment of cough, fever, chronic hepatitis, gastric ulcer, liver and
breast cancer food poisoning and as a general tonic (Chang and Lee, 2004; Wong et al, 2009; Lee et al., 2009).
Even our former prime minister, Tun Dr. Mahathir, during his opening speech at Biomalaysia, 2002 mentioned
that his chronic cough had been cured by this medicinal mushroom. In China, L. rhinocerotis has also been used
in the traditional Chinese medicine to treat liver cancer, chronic hepatitis and gastric ulcer (Huang, 1999).
Research findings have shown that the sclerotial polysaccharides from the mushroom demonstrate very high
anti-inflammatory, antioxidant, anti-proliferative (Lai et. al. 2008) and immuno-modulating effects (Wong et. al.,
2009; Wong et. al., 2010; Guo et. al., 2011).
L. rhinocerotis is structurally characterised to have a central stipitate pilei arising from distinct sclerotia
(Ryvarden and Johansen, 1980). The sclerotium of L. rhinocerotis is the part with medicinal value. Its
geographical distribution is only in the tropical rainforest in the region of Thailand, Malaysia, Indonesia,
Philippines, and Papua New Guinea. The existence of this mushroom in the jungle is always solitary and this
makes the collection of its sclerotia a difficult task. As a result, its supply is limited and the sclerotium is costly.
Recently, we reported a breakthrough in the cultivation of the Lignosus rhinocerotis strain, LiGNO™ TM02 on
agar, solid and spawn medium with high production yield (Tan et al., 2009), thus overcoming the supply problem.
The nutrition industry is ever growing, despite poor economic outlook. The market for functional foods remains
bullish. In this modern society, functional food demandis driven by the desire for products that help build and
maintain optimum health. Functional foods contain special ingredients with unique beneficial effects from
cardiovascular to mental health function, immunity. The list of contemporary health issue of public concern is
growing rapidly. The continuous demands for nutraceuticals create new opportunities and commercial pressures.
Hence, the nutrition industry is always looking for potential functional ingredients. The cultivated L. rhinocerotis
has a huge potential to be exploited in this arena.
Although L. rhinocerotis is recognized as a rare species, it still can be found in the jungle. The study started with
L. rhinocerotis specimens collected from the forest in Cameron Highland, Sungai Perak, Gerik, Hulu Langat, and
Raub. The spore and the tissue from its sclerotium, stem and pileus were grown in a special formulated culture
media. The mycelium growth was then subjected to many cycles of sub-culturing in order to obtain a clean, pure
culture. Genetic marker was developed in order to authenticate the specimens collected (Tan et al, 2010). The
L.rhinocerotis culture growth conditions in liquid and solid media were optimised. Since the sclerotia of
L.rhinocerotis is the part that contained medicinal properties, cultivation factors and conditions for the formation
of sclerotia were also determined and optimised. The cultivation process takes approximately 6 months (for the
sclerotia formation) to up to 2-3 years (for stem and pileus formation).
At present, we are able to produce the sclerotia of L.rhinocerotis at commercial scale in an environmentally
controlled culture room. The production is about 100 kg per month and can easily be expanded by increasing the
parameters of its culture room.
In the development of any functional ingredient, the safety has to be established based on its historical use, its
intrinsic nature, usage or based on information generally known and accepted by qualified scientific experts.
Although history reports that L. rhinocerotis has been extensively used safely for over hundreds of years, a
scientific assessment remains essential. Preliminary toxicity study showed that oral administration of the
cultivated sclerotial powder at daily dose of up to 10% of the experimental animals’ body weight continuously for
three months did not show any adverse effects. Various toxicity studies in compliance with the OECD
guidelines were subsequently conducted.
Acute toxicity study showed that there was no treatment-related acute toxicities in rats following oral
administration at a high dose of up to 2000 mg/kg. 28 days sub-acute toxicity study (Lee et. al., 2011) showed
that oral administration of the sclerotial powder at daily dose of up to 1000 mg/kg had no adverse effects on the
growth rate, hematological and clinical biochemical parameters. Histological studies showed that the treatments
did not induce any pathological changes in the liver, kidney, heart, spleen and lung of the animals. As the
highest tested dose of 1000 mg/kg was not associated with any toxicity concerns, the NOAEL dose is concluded
to be higher than 1000 mg/kg.
Teratogenicity study showed that the oral admistration of 100mg/kg sclerotial powder for 7-8 weeks did not
significantly (p>0.05) alter the fertility of rats and did not induce teratogenic effect on their offspring. Besides that,
external sign of malformation was not observed in all pups from treated as well as control group. Besides, the
mutagenicity study has also proven the cultivated L. rhinocerotis is non-mutagenic in the bacterial reverse
mutation assay.
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The cultivated L.rhinocerotis was also subjected to corticosteroid screening by Toxicology Laboratory, National
Poison Centre. The results confirmed the absence of corticosteroid as part of its ingredient.
The chronic toxicity toxicity study is currently ongoing.
Continuous research on the bioactive properties of L. rhinocerotis is essential to generate proof to substantiate the
ethno-botanical claims of L. rhinocerotis and to support its contentst with evidence-based proofs.
Anti-inflammatory: The present study investigates the anti-inflammatory activity of the sclerotia of the
cultivated L. rhinocerotis using carrageenan induced anti-inflammatory model (rat paw edema). The cold water
extract has shown significant anti-inflammatory effects comparable to standard drug, indomethacin (unpublished
Immuno-modulating: Sclerotial polysaccharides of L. rhinocerotis have been associated in animal studies with
multiple bioactive properties. Extracts of L. rhinocerotis, and particularly the sclerotial polysaccharide
constituents have shown stimulatory effects on human innate immune cells. The constituents have also been
associated with immune modulation in preclinical study, and are hypothesised to exert anti-tumor effects as a
result of these immune properties (Wong et. al., 2009; Wong et. al., 2010; Guo et. al., 2011).
Anti-Oxidant : The findings from a recent in vitro study (unpublished data) suggest that the antioxidant capacity
of the mushroom sclerotium is comparable to many other medicinal mushrooms, which is generally moderately
low. The water extracts, however, exhibited strong superoxide anion scavenging activity, indicating that the
extract might be helpful in preventing certain type of oxidative stress. Taken as a whole, the L.rhinocerotis
sclerotial powder offer a promising source of functional ingredient potentially attributed to its antioxidant capacity,
specifically the superoxide anion scavenging activity.
Anti-proliferative : Lai et. al. (2008) was the first to investigate the anti-proliferative effects of the sclerotial
polysaccharides of the mushroom. They reported that the hot water extract of P. rhinocerotis (synonym of L.
rhinocerotis) exhibited anti-proliferative activity against different kinds of leukemic cells.
Recently, Lee et. al. (2012, in press) reported that the cold water extract of the sclerotia of the cultivated L.
rhinocerotis exhibited significant anti-proliferative activity against the breast cancer cell MCF-7 and lung cancer
cell A549. Their results also showed that the cold water extract was essentially not cytotoxic against the normal
breast and lung cells. Its cytotoxicity action is due to a high molecular weight fraction isolated from the cold
water extract, and that the cytotoxic action is mediated via apoptosis. The anti-proliferative action against
MCF-7 cells provides a plausible scientific basis for the traditional use of L. rhinocerotis sclerotia in breast cancer
treatment by the Malaysian indegenous communities.
Traditional medicine is a system of medicine based on cultural beliefs and practices handed down from
generations to generations. The practice continues until today and the World Health Organization (WHO)
estimates that 65 - 80% of the world’s population rely on traditional medicine for their primary health care needs.
Regulation of traditional medicine in Malaysia began in 1992 and applications for product registration have to be
submitted to the National Pharmaceutical Control Bureau (NPCB). Only ingredients listed in the NPCB
Traditional Medicine Active Ingredients List are eligible to be used in any registered products. Unfortunately L.
rhinocerotis was not in the list prior to September 2010. In order to qualify as functional ingredient, we have
submitted to NPCB the supporting documents on the safety, history of use and bioactivities of L. rhinocerotis.
NPCB had accepted L. rhinocerotis to be listed in the traditional medicine active ingredient list in September 2010.
This enables L. rhinocerotis to be exploited commercially as a functional ingredient.
The full potential of L. rhinocerotis high in medicinal and nutritional values has never been realised due to its
limited supply. Now with the successful cultivation technology at commercial scale, the supply problem is a thing
of the past. In order to further qualify as a functional ingredient, evidencebased safety and efficacy assessment is
crucial and essential.
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Study on traditional Chinese medicinals
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