BAIF Development Research Foundation

ETHNOVETERINARY MEDICINE: ALTERNATIVES FOR LIVESTOCK DEVELOPMENT
PROCEEDINGS OF AN INTERNATIONAL CONFERENCE HELD IN PUNE, INDIA, 4-6 NOVEMBER 1997
VOLUME 1: SELECTED PAPERS
FILE 4 OF 9: PART 3: ETHNOVETERINARY MEDICINAL PLANTS AND PLANT MEDICINES


Key words and phrases:

animal health, community based animal health care, environment, ethnoveterinary medicine, indigenous knowledge, indigenous systems, participation, veterinary

Edited by:
Evelyn Mathias
D.V. Rangnekar
and Constance M. McCorkle
with the assistance of
Marina Martin

Published 1999 by BAIF Development Research Foundation, Pune, India 1999

BAIF Development Research Foundation
BAIF Bhavan, Dr. Manibhai Desai Nagar
Warje Malewadi (Bombay - Bangalore bypass highway)
Pune 411 029, India
Phone +91-212-365 494, fax: +91-212-366 788

BAIF is a non-political, secular non-governmental organisation involved in livestock development. BAIF's mission is to create opportunities of gainful self-employment for rural families, especially disadvantaged sections, ensuring sustainable livelihood, enriched environment, improved quality of life and good human health. This will be achieved through development research, effective use of local resources, extension of appropriate technologies and upgradation of skills and capabilities with community participation.

Correct citation:
Mathias, E., D.V. Rangnekar, and C.M. McCorkle. 1999. Ethnoveterinary Medicine: Alternatives for Livestock Development. Proceedings of an International Conference held in Pune, India, on November 4-6, 1997. Volume 1: Selected Papers. BAIF Development Research Foundation, Pune, India.


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CONTENTS
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Part 3: Ethnoveterinary medicinal plants and plant medicines

Ethnomedico-botany and its sustenance
S. Vedavathy

Less-known ethnoveterinary uses of plants in India
R. L. S. Sikarwar

Evaluation of indigenous herbs as antitrypanosomal agents
S. K. Dwivedi

The potential of Tinospora rumphii as an anthelmintic against H. contortus in goats
Tomas. J. Fernandez

Therapeutic efficacy of a herbal preparation in dermatological conditions of animals and its influence on wound healing
B. P. Manjunatha

Zeetress - a promising anti-stress remedy and immuno-modulator. A review
C. B. Pande


PART III: ETHNOVETERINARY MEDICINAL PLANTS AND PLANT MEDICINES


Ethnomedico-botany and its sustenance

S. Vedavathy

Introduction

Ethnomedico-botany focuses on the knowledge of medicinal plants that people have developed over generations. The World Health Organisation estimates that as many as 80% of the world's population depend on plants for their primary healthcare (Farnsworth et al. 1985).

Conservation of biodiversity

India is one of the World's 12 regions having the largest biodiversity. It has 45000 plant species of which 15000-20000 possess proven medicinal value (Krishna Kumar 1996).

It is estimated that world-wide about 24 ha of rain forest disappear every minute. This destruction also reduces the supply of medicinal plants many of which grow in forests. We have therefore to find ways to overcome the decreasing availability of herbs, especially as the demand is increasing. The following actions are needed:

  • Studies of plant taxonomies have to be updated. Biodiversity can be assessed and utilised only after determining the present state of the local and regional flora.
  • A network of protected areas should be established to prevent the loss of the rich natural wealth. This would also help to preserve a range of ecotypes and a gene pool of medicinal plants.
  • For many wild species of medicinal plants, no suitable cultural practices are known. Therefore the domestication of species should be studied, especially of those that are under pressure. Biotechnology can help in the propagation of endangered species though it does not invent but uses already existing genes.
  • There is an urgent need to speed up programmes to inventorise plants and screen them for pharmacological activity.

Conservation of ethnomedico-knowledge

In India, ethnomedico-botanical surveys and inventories receive little attention and detailed information and documentation on the uses of medicinal plants in indigenous communities are lacking. This disinterest in the existing knowledge and wanton neglect by authorities and public slowly result in the loss of this knowledge. It is high time that effective measures and programmes are adopted to save this information before it is lost for ever. Ethnomedico-botanical surveys in each district are needed (Arora 1997). Comparing and cross-checking the results will provide us with information about herbal medicines presently in use. Furthermore, concerted efforts are needed towards the usage of plant resources, linking conservation strongly with utilisation.

National health policy for indigenous medicine

Indigenous medical systems have been and still are playing a major role in healthcare. However, traditional healthcare presently receives only about 5% of the national budget in India. This lack of support excludes the indigenous systems from mainstream healthcare, i.e., western medicine, and hampers its development.

Indigenous healthcare systems should be included into the curricula of medical colleges and teaching hospitals dealing with modern medicine. One has to emulate the Chinese in this respect. To understand and encourage indigenous systems of healthcare should not be viewed as a potential threat to modern medicine. No system is faultless and each has positive aspects. An amalgamation of the different systems is needed to foster a sound national health policy.

Collaboration between indigenous medical systems

The lack of communication between indigenous medical systems of different countries is a major reason why the systems are not able to convince the world of its rich traditions and sound principles. For example, there are many commonalties between India and China. Both countries have cultural histories dating uninterrupted back thousands of years. Actually, if the countries of South and Southeast Asia which have common knowledge systems come together to discuss these aspects, much hidden information on indigenous systems could be unearthed.

Rights of indigenous people

In early civilisations food and medicine were linked and many plants were eaten for their therapeutic properties. The knowledge, resources, and rights of ethnic groups have been continuously ignored. We now have to represent their aspirations and needs. We always consider knowledge as a public property belonging to the society and the individual is considered as a temporary carrier. In indigenous societies, the knowledge is passed on from mother to daughter, father to son and teacher to disciple. Abuse and misuse of knowledge is regarded as a great sin and commercialisation also has certain norms, to be taken with the consent of indigenous people.

The knowledge base of indigenous peoples is now facing the threat of alienation through us outsiders. We may have to bring it back from the west paying exorbitant fees if the present conditions prevail. It is the duty of authorities and intellectuals to take appropriate steps to protect the knowledge and rights of indigenous peoples and prevent the loss of natural resources and exploitation of knowledge.

Paramedicines and patent rights

The World Trade Organisation controls the international trade relations. For intellectual property rights, a patent war is emerging and developing countries like India should take up the challenge.

Indigenous medical systems are relying on crude herbal drug preparations. We have to adopt modern analytical methods and subject the drugs to enhanced quality control. This does not mean that traditional drugs should strictly adhere to the standardisation on par with chemical drugs. It is not possible to compare herbal drugs with chemical ones because most of the herbal medicines are mixtures of numerous chemical molecules.

But it is necessary to develop standard specifications for herbal medicines by indicating the ingredients, the amount and range of the active principles, their therapeutic properties, etc. Better methods to improve the shelf life can be achieved with the aid of modern analytical instruments (Natesh 1997). Products of improved quality will certainly help a large section of the population that depends on traditional products. Perhaps India may be even able to sell such products in the western market (Bhatia 1997).

The Food and Drug Administration of the USA has been maintaining that herbal formulations should be grouped only as ‘food supplements.' In that case herbal medicines belong neither to medicines nor to food. According to western norms, chemical drugs can be subjected to standardisation since they contain known compounds while herbal drugs are mixtures of natural products and contain numerous chemical molecules. The synergistic action of ingredients is considered only in connection with preparation but not for the individual constituents. In fact, attempts to isolate the active principles failed in a number of plants despite the usage of new techniques in natural product analysis.

Traditional medicines can be prepared under enhanced quality control without sacrificing the therapeutic quality. It is difficult to go for this type of analysis in the beginning but it becomes a magic wand whereby a new class of formulations called ‘paramedicines' (neither food nor medicine) can be released into the international market soon. Having sound research and development support is certainly going to be advantageous to manufacturers. The emergence of drugs like Memory+ from Becopa monnieri marks the beginning of this type of approach. Many more such drugs should come soon and the industry should wake up and get ready to become global players.

References

Arora, R. K. 1997. Ethnobotany and its role in the conservation and use of plant genetic resources in India. Paper presented during the Training Course on Ethnobotany at S. V. University, Tirupati, India, on 17 March 1997.

Bhatia, C.R. 1997. Standardisation of herbal products. Paper presented during the Pre-project Formulation Workshop on Biotechnological Interventions for Improvement of Medicinal and Aromatic Plants held at the Regional Agricultural Research Station, Palem, Mahaboob Nagar, A. P., India.

Farnsworth, N. R., O. Akerele, A.S. Bingel, D. D. Soejarta, and Z. Eno. 195. Medicinal plants in therapy. Bulletin of the World Health Organization 63(6):965-981.

Krishna Kumar, P. R. 1996. Indian medicine industry under the Emerging patent regimes. Ancient Scinece of Life 15(3):161

Natesh, S. 1997. Biotechnological interventions in conservation and improvement of medicinal and aromatic plants. Paper presented during the Pre-project Formulation Workshop on Biotechnological Interventions for Improvement of Medicinal and Aromatic Plants held at the Regional Agricultural Research Station, Palem, Mahaboob Nagar, A. P., India.


Less-known ethnoveterinary uses of plants in India

R. L. S. Sikarwar

Introduction

The value of studies comparing the ethnobotany of distant and distinct regions within a country or of different countries is universally recognised now. Such comparative studies yield valuable information on the uses and properties of plants and the width and depth of indigenous knowledge. Common plant uses by distantly located and distinct ethnic groups point to a similar evolution of indigenous knowledge and the credibility of the plants' medicinal value. The information on plant uses widely practised in one location could be very useful to other places having the same plants but not using them the same way.

In 1991 a meeting of scientists of Asian and Latin American countries was held at Belem in Brazil under the United Nations Development Programme (UNDP). It was unanimously decided to exchange information on indigenous knowledge about plants from these regions to improve and enhance the utilisation of plant resources. In this context UNDP supported a small one-year project on ‘Comparative Ethnobotanical Studies between India and Amazonia (Brazil region)' in 1993-1994.

In India the Council of Scientific and Industrial Research (CSIR) realised the importance of comparative studies. It supported a project under the Emeritus Scientist Scheme of Dr. S.K. Jain (Jain 1991, Jain et al. 1991) and an elaborate research work ‘Cross-cultural Ethnobotanical Studies of Northeast India' to identify the common plants different tribal communities of Northeast India use to reduce or alleviate various diseases (Jain and Saklani 1992, Saklani and Jain 1994, 1996). Later it funded ‘Studies on Comparative Ethnobotany of India and Latin America (LA) - Search for Underutilized Bioresources' to identify new plant resources for utilisation in India and Latin America.

The comparative studies identified about 650 plants common to both Latin America and India. Of these, 259 plants used as folk medicines were documented in the well-known literature and published papers from India (Ambasta 1986, Anonymous 1948-76, Chopra et al. 1956, Jain 1991, Singh et al. 1983) and Latin America (Balbach 1980, Correa 1926-1976, Di Stasi et al. 1989, Duke 1986, Duke and Vasquez 1994, Lorenzi 1991, Rodrigues 1989, Schultes and Raffauf 1990, Standley 1920-1926).

The analysis of the uses of these plants showed that some medicinal uses were common to both regions, i.e., the same plant or plant part was used for the same disease or different parts of the same plant were used for the same disease, while certain medicinal uses appeared less known or unknown in India but were extensively applied by people in Latin America and vice versa. An account of common uses of plants of both regions (Jain et al. 1995) and some uses that appeared less known or unknown to India (Jain and Lata 1996, Jain and Sikarwar 1996) have been published.

Latin America plant uses not practised in India can be very useful to India which is mainly an agricultural country; about 80% of its population reside in rural areas. Indian livestock keepers raise domestic animals such as cows, oxen, buffaloes, sheep, goats, horses, camels, dogs, fowls, etc., for milk, food, agriculture, and markets. There are only a few veterinary hospitals. They are located at district and block levels far from rural and remote areas. Therefore many people treat their sick animals through herbal medicines that are locally available but vary seasonally. More than three dozen papers on ethnoveterinary medicines have been published from different parts of India.

Table 1 describes the ethnoveterinary uses of 17 plants from Latin America less known in India. A scrutiny of the literature indicates that these uses are so far not prevalent in India and merit further study.

Our studies on other aspects of comparative ethnobotany are continuing and we hope to identify many more less known potential uses of Indian plant resources.

Acknowledgements

The author is thankful to Dr. S. K. Jain, Director, Institute of Ethnobiology, for encouragement and valuable suggestions; the Director of the National Botanical Research Institute, Lucknow for providing facilities; and The Council of Scientific and Industrial Research, New Delhi, for financial assistance.

References

Ambasta, S. P. (ed.). 1986. The Useful Plants of India. Publication and Information Directorate, Council of Scientific and Industrial Research, New Delhi, India.

Anonymous. 1948-1976. Wealth of India (Raw Materials). Council of Scientific and Industrial Research, New Delhi, India.

Balbach, A. 1980. A flora nacional na medicina domestica (vol.2). Itaquacetuba, Sao Paulo, Brazil.

Chopra, R. N., S. L. Nayar, and I. C. Chopra. 1956. Glossary of Indian Medicinal Plants. Council of Scientific and Industrial Research, New Delhi, India.

Correa, M. P. 1926-1976. Dicionario das plantas uteis do Brasil. Imprensa Nacional, Rio de Janeiro, Brazil.

Di Stasi, L. C. et al. 1989. Plantas medicinais na Amazonica. UNESP, Sao Paulo, Brazil.

Duke, J. A. 1986. Isthmian Ethnobotanical Dictionary. Scientific Publishers, Jodhpur, India.

Duke, J. A. and R. Vasquez. 1994. Amazonian Ehnobotanical Dictionary. CRC Press, Inc. Boca Raton, Florida, USA.

Jain, S. K. 1991. Dictionary of Indian Folkmedicine and Ethnobotany. Deep Publications, New Delhi, India.

Jain, S. K. and Sneh Lata. 1996. Unique indigenous Amazonian uses of some plants growing in India. Indigenous Knowledge and Development Monitor 4(3): 21-23.

Jain, S. K. and A. Saklani. 1992. Cross-cultural ethnobotanical studies in northeast India. Ethnobotany 4:25-38.

Jain, S. K. and R.L.S. Sikarwar. 1996. Comparative ethnobotanical studies between India and Amazona- a new field of research [in Hindi]. Sacchitra Ayurved 49(2):96 and 155-159.

Jain, S. K., B. K. Sinha, and R. C. Gupta. 1991. Notable Plants in Ethnomedicine of India. Deep Publications, New Delhi, India.

Jain, S. K., V. F. Farnandes, Sneh Lata, and A. Ayub. 1995. Indo-Amazonian ethnobotanic connections - similar uses of some common plants. Ethnobotany 7:29-37.

Lorenzi, H. 1991. Plantas daninhas do Brasil. Nova Odessa, Brazil.

Rodrigues, R. M. 1989. A flora da Amazonia. CEJUP, Belem, Brazil.

Saklani, A. and S. K. Jain. 1994. Cross-cultural Ethnobotany of Northeast India. Deep Publications, New Delhi, India.

Saklani, A. and S. K. Jain. 1996. Credibility of folk claims in Northeastern Himalaya and Northwestern India. In: S. K. Jain (ed.). Ethnobiology in Human Welfare. Deep Publications, New Delhi, India. Pp.136-139.

Singh, U., A. M. Wadhwani, and B. M. Johri. 1983. Dictionary of Economic Plants in India. Indian Council of Agricultural Research, New Delhi, India.

Schultes, R. E. and R. F. Raffauf. 1990. The Healing Forest. Dioscorides Press, Portland, Oregon, USA.

Standley, P. C. 1920-1926. Trees and Shrubs of Mexico. Smithsonian Press, Washington, D.C., USA.

Table 1. Ethnoveterinary uses of plants in Latin America less known in India.

Scientific plant name (plant family) ‘Hindi plant name'

Plant description

Use in Latin America (reference)

Allium sativum L. (Liliaceae) ‘lasan'

Bulbous annual herb

Bulb used against fowl diseases (Correa 1926-1976)

Annona squamosa L. (Annonaceae) ‘sarifa'

Small tree with greenish yellow flowers

The leaves are sometimes rubbed over floors or placed in hens' nests to keep away vermin (Standley 1920-1926)

Bixa orellana L. (Bixaceae) ‘latkan'

Small tree with white flowers

In Brazil the pulp of the seeds is given to bulls before fights to make them more active and ferocious. The plant may contain some excitant which has not yet been investigated (Standley 1920-1926)

Caladium bicolor Vent. (Araceae)

A rhizomatous herb

Peasants use leaf decoction to get rid of external cattle festers caused by worms (Balbach 1980)

Capsicum annuum L. (Solanaceae) ‘mirch'

Annual herb with white flowers

Curanderos use it as a maceration mixed with aguardiente (liquor) to cause purging in dogs, so as to make them good hunting dogs (Duke and Vasquez 1994)

Capsicum frutescens L. (Solanaceae) ‘mirch'

Annual herb with white flowers

Creole people use it for throat diseases of pigs (Duke and Vasquez 1994)

Cassia tora L. (Caesalpiniaceae) ‘panvar'

Annual herb with yellow flowers

If the juice of mashed leaves is given to an animal, the ticks will jump off. This anti-tick folklore seems exaggerated (Duke and Vasquez 1994).

Ceiba pentandra (L.) Gaertn. (Bombacaceae) ‘safed semal'

Medium-sized tree with greenish flowers

An infusion of the bark is given to cattle after delivery to help expel the placenta (Duke 1986)

Chenopodium ambrosioides L. (Chenopodiaceae)

An erect aromatic herb with greenish or purplish flowers

Leaves are used to expel worms in animals (Di Stasi et al. 1989)

Crescentia cujete L. (Bignoniaceae) ‘bilayati bel'

Medium-sized tree with campanulate flowers

Cattle eat the fruit often during the dry season but it is said that it often causes abortion (Standley 1920-1926)

Leucaena glauca Benth. (Mimosaceae) ‘subabool'

Medium-sized tree with whitish globular heads

There is a prevalent belief that if horses, mules or pigs eat any part of the plant, their hairs will fall out. Cattle are said not to be affected (Standley 1920-1926)

Luffa acutangula (L.) Roxb. (Cucurbitaceae) ‘kalitori'

A climbing or trailing annual with yellow flowers

Unripe fruit is used against bowel disease of domestic fowl (Correa 1926-1976)

Table 1 (continued)

Scientific plant name (plant family) ‘Hindi plant name'

Plant description

Use in Latin America (reference)

Luffa aegyptiaca Mill. (Cucurbitaceae) ‘ghiya tori'

A climbing or trailing annual with yellow flowers

Fruits are used the same way as L. acutangula (Correa 1926-1976)

Mammea americana L. (Cluciaceae)

Small or medium-sized tree with white fragrant flowers

The gum obtained from the bark is used to extract chiggers from the skin and kill ticks and other parasites external parasites of domestic animals (Standley 1920-1926)

Nicotiana tabacum L. (Solanaceae) ‘tambaku'

Viscid annual herb with rosy or reddish flowers

Powdered tobacco is mixed with aguardiente (liquor) and given to dogs to make them better hunting animals (Duke and Vasquez 1994)

Paspalum conjugatum P. Bergius (Poaceae)

Annual grass

Palikur people use it with other plants to prepare hunting dogs (Duke and Vasquez 1994)

Petiveria alliacea L. (Phytolaccaceae)

An erect herb with rosy or white flowers

People believe when cows eat the plant, their milk will have an onion-like flavour (Standley 1920-1926)


Evaluation of indigenous herbs as antitrypanosomal agents

S. K. Dwivedi

Introduction

Indian Ayurveda is a one of the noteworthy systems of traditional medicine practice that uses mainly medicinal plants for the treatments of ailments in both people and animals. Although the popularity of herbal medicine recorded a sharp decline after the introduction of allopathic chemical drugs, herbal medicines are gaining growing interest because of their cost-effective and eco-friendly attributes. Recent observations indicate that perhaps 80% of the world's population rely solely upon medicinal plants for the treatment of diseases. Furthermore, a major part of chemically synthesised drugs against infectious agents is in fact derived from natural products or from structures suggested by natural products (Kirby 1996).

Trypanosomiasis, also known as ‘surra', is one of the economically most important diseases of farm animals affecting livestock health and economy in several tropical countries including India. Lack of cost-effective drugs is a serious drawback in the treatment of this disease. Most of the antitrypanosomal drugs currently available in the market are either highly toxic to animals or the parasite rapidly becomes resistant to these drugs (Mansfield 1984, Nantulya and Moloo 1989, Williamson 1976). A potent trypanocidal drug without side effects is therefore urgently needed. Virtually no new antitrypanosomal allopathic drugs for commercial purposes have been introduced over the past 40 years which clearly underlines the need to find novel antitrypanosomal agents, particularly those with a wide safety margin and less or no undesirable side effects. One difficulty in the development of antitrypanosomal drugs is the fact that protozoa, unlike other pathogens, share many metabolic pathways with their mammalian hosts. Therefore we need to find compounds that selectively kill the protozoa while affecting the host's cellular metabolism as little as possible. Medicinal plants can be an answer to this problem.

Principles of the evaluation of antitrypanosomal herbs

Research on medicinal plants as a source of antitrypanosomal drugs calls upon both pharmacognosists and clinical parasitologists. The plants are mainly selected on the basis of their traditional reputation for efficacy in the treatment of trypanosomiasis and other disease. Selected plants are subjected to preparation and/or purification of extracts. Initially it is imperative to go for in vitro primary screening which reduces the number of laboratory animals used for experiments. Although agents active in vitro are often inactive in vivo and vice versa, the in vitro system can act as a primary screen and helps to identify plants for in vivo testing. If a compound kills the parasite in vitro, it will also be screened for toxicity against mammalian cells in vitro. A plant's active ingredients are isolated and identified with chromatography, mass spectrometry, nuclear magnetic resonance and other techniques (Kirby 1996). Identified agents are then further tested for their efficacy and toxicity in vitro.

In vivo studies are done in an animal model, preferably mice infected with trypanosomes. The activity of the test material in vivo is influenced by a number of factors: The compounds effective in vitro may not be effective in vivo due to their failure to reach the requisite site of action or they metabolise too quickly to a less active or inactive form. Or a compound can be more active in vivo because it gets metabolised to a more active form. For example, Berberis alkaloids have little activity in vitro but are activated by metabolism in animal models to become active against Entamoeba histolytica (Phillipson et al. 1993).

When a medicinal plant proves effective in vivo and shows no host toxicity, its mechanism of killing parasites is studied through complex and extensive biochemical testing. Also, a likely effect on the host's metabolism must be studied before a drug is released for extensive clinical and field trials. It is also desirable to find out whether a drug has broad spectrum activity by testing it against a range of alike organisms.

Test protocol

Extensive in vitro and in vivo trials have been conducted at our laboratory to screen 23 indigenous medicinal plants for their trypanocidal potential (Table 1). These plants were selected because of claims that they possess antiprotozoal activity and alleviate one or many of the clinical symptoms such as intermittent fever, anaemia, jaundice, and hepatomegaly commonly associated with trypanosomiasis. These herbs were tried at different concentrations against a pathogenic strain of Trypanosoma evansi, isolated from a clinical case of trypanosomiasis and maintained in the laboratory by successive serial passages in mice. The parasitic inoculum was prepared by collecting 1-2 drops of blood from a mouse showing severe (++++) parasitaemia and diluting it in Alsever's solution. For both in vitro and in vivo testing mice were inoculated intraperitoneally with 106 organisms per mouse. The trypanosomes were separated from the heart blood of the inoculated mice for in vitro testing which was performed by the two dimensional method using micro test plates. The antitrypanosomal activity was assessed on the basis of change in number, motility, and infectivity of the parasite in mice. For the in vivo studies, three different doses of the test drug, selected on the basis of LD50 and a minimum therapeutic dose (MTD), were given intraperitoneally to mice which had developed parasitaemia following experimental inoculation with 104 trypanosomes intraperitoneally. The animals were observed for 30 days to record pattern of parasitaemia, survival rate, and mean survival period in inoculated mice. Triquin (a standard allopathic trypanocidal commercial preparation) was used to compare the efficacy. The herbal drugs found to possess antiprotozoal activities in vitro and in vivo were tested for their toxicity in rats at the highest dose concentrations given for five consecutive days or up to the death of animals.

Results and discussion

Of the 23 plants tested, fresh juice and aqueous and alcoholic extracts of Xanthium strumarium leaves and Parthenium hysterophorus flowers, aqueous and alcoholic extracts of Nyctanthes arbortristis leaves, and alcoholic extracts of Aristolochia indica stems revealed 100% trypanocidal activity in vitro (Table 2). The alcoholic extracts of Xanthium strumarium leaves, Parthenium hysterophorus flower and Nyctanthes arbortristis leaves were also found effective in vivo at dosages of 100 and 300 mg/kg body weight. Aristolochia indica, however, while 100% effective in vitro, did not reveal any antitrypanosomal activity when tested in infected mice.

Other plants exhibiting mild to moderate trypanocidal activity in vitro included root, bark, and leaves of Azadirachta indica, leaves of Cassia occidentalis and Hydrocotyle asiatica, rhizomes of Cyperus rotundus, seeds of Cannabis indica, stem bark of Holarrhena dysenterica and leaves of Ocimum sanctum. However, except the ethanolic extract of Canabis indica seed which produced a mild to moderate activity, none of these plants exhibited antitrypanosomal activity in vivo.

The toxicity trials revealed that 50% ethanolic extract of Xanthium strumarium leaves was hepatoxic and killed 50% of the mice within 4-12 days when given at doses of 1000 mg/kg body weight intraperitoneally. At a similar dose and administration route, 50% ethanolic extract of Parthenium hysterophorus flowers killed all the six test mice within four hours. The alcoholic extract of Nyctanthes arbortristis leaves was less toxic and caused 33.4% mortality in rats besides enhancing serum aspertate aminotransferase and alanine aminotransferase values within 12 days of the observation period.

The 23 plants tested differed in their antitrypanosomal activity. Only the alcoholic extracts of three plants, i.e., Xanthium strumarium leaves, Parthenium hysterophorus flowers, and Nyctanthes arbortristis leaves, were found effective both in vivo and in vitro. The other plants were not active in vivo.

The plants' antitrypanosomal activity could be due to alkaloids or other ingredients having antiprotozoal activity in vitro. However, when used in vivo, the antitrypanosomal activity did not become apparent. This could be due to degradation or metabolisation of the active principle through various metabolic processes in the host animal. This means that extracts that are effective in vitro are not necessarily active also in vivo. Therefore extracts should be tested both in vitro and in vivo.

The three extracts that were effective in both in vitro and in vivo were toxic to the host. This could be due to the fact that Trypanosoma and the host animal are both eukaryotic (i.e., have cells with a true nucleus) and have similar metabolic pathways. The extracts that are effective in both in vitro and in vivo should be fractionated to determine the active principle within the biological system.

Conclusion and future strategies for validation

No medical system alone can fully address all challenges and complexities of health problems of the modern period. Practitioners of the different medical systems should therefore collaborate and pool their knowledge to their mutual advantage and apply whatever is the best from the various healthcare systems. This principle can also be applied to find a new antitrypanosomal agent.

The introduction of artemisinin, a plant-derived antimalarial compound, is a good example of how medical systems based on herbal medicines can be tapped in identifying new antiprotozoal drugs. It is possible that a plant-derived medicine might be effective in cases of chronic and recurring surra where the hosts have some degree of immunity. Certain plants indigenous to India have shown antitrypanosomal potential in vitro and in vivo. These plants can be further evaluated and validated for use.

However, we have to decide our validation strategies according to the epizootiological profile of trypanosomiasis in India which obviously is quite different from western countries where the disease occurs only sporadically. Therefore developed countries with such a low disease profile may prefer a drug which can kill the trypanosome. In tropical countries, on the other hand, where trypanosomiasis is epizootic, a plant-derived drug controlling the growth of the parasites while having antipyretic and possibly other palliative properties are also acceptable.

Because the host's immunity plays an important role in combating pathogens, a herbal agent which allows the build-up of immunity might prove beneficial while an agent suppressing immunity may be deleterious. Therefore it is worthwhile to test plant materials with trypanocidal potential also for their immunomodulatory effects.

Testing of mixtures of various indigenous herbs may be an appropriate validation approach to develop a novel cost-effective drug. The herbs for such mixtures would have to be selected carefully on the basis of their antiprotozoal activity and potential to alleviate clinical signs produced by trypanosome together with immunostimulatory activity.

Traditional healers in India have a vast treasure of knowledge on herbalism which should be validated scientifically. Efforts to document this knowledge need to be enhanced but this will be only possible if ethnopharmacologists and traditional practitioners establish a close rapport. Scientific validation should investigate how to maximise and standardise the efficacy of traditional remedies. We should also attempt to understand the active principles, look for evidence of synergism and find out whether a defined mixture of plant-derived compounds can replace the use of the crude herbal preparation. Finally, we should promote conservation of medicinal plants in order to provide cost-effective eco-friendly drugs against tropical diseases such as trypanosomiasis in farm animals. This could help to save many medicinal plant species from becoming extinct.

Reference

Kirby, G. C. 1996. Medicinal plants and the control of protozoal diseases, with particular reference to malaria. Transactions of the Royal Society of Tropical Medicine and Hygiene 90:605-609.

Mansfield, J. M. (ed.). 1984. The Chemo Therapy (vol. 2). Marcel Dekker Inc, New York. P. 233.

Nantylya, V. M. and S. K. Moloo. 1989. Recent developments in trypanosomiasis. International Journal of Animal Science 4:71-84.

Phillipson, J. D., C. W. Wright, G. C. Kirby, and D. C. Warhurst. 1993. Tropical plants as sources of antiprotozoal agents. Recent Advances in Phytochemistry 27:1-40.

Williamson, J. 1976. Chemotherapy of African trypanosomiasis. Transactions of the Royal Society of Tropical Medicine and Hygiene 70:117-119.

Table 1. Medicinal plants tested for trypanocidal activity in vitro and in vivo.

Plant name in Latin and Hindi or English

Parts and preparation used

Doses used (m g/ml)

in vitro in vivo

Achyranthes aspera (apamarga)

Aqueous extract of dried leaves

5, 50, 500, and 1000

-

Aristolochia indica (isharmul)

80% ethanolic extract of dried stem

100, 500, and 1000

180, 125, 95

Azadirachta indica (neem)

80% ethanolic extract of fresh root bark

100, 500, and 1000

340, 227, 170

 

Aqueous extract of dried leaves

5, 50, 500, and 1000

-

Caesalpinia bonducella (kat karanj)

Aqueous extract of dried seed

5, 50, 500, and 1000

-

Cannabis indica (bhang)

80% ethanolic extract of dried seed

100, 500 and 1000

250, 170, 125

Calotropis procera (madar)

80% ethanolic extract of fresh root

100, 500 and 1000

275, 183, 137

Cassia occidentalis (kasondi)

Aqueous extract of dried leaves

5, 50, 500, and 1000

-

Cissampelos pareira (harjori)

80% ethanolic extraxt of dried whole plant

100, 500, and 1000

500, 335, 250

Cyperus rotundus (motha)

Aqueous extract of dried rhizomes

5, 50, 500, and 1000

-

Datura alba (sodah-dhatura)

Aqueous extract of dried leaves

5, 50, 500, and 1000

-

Eclipta prostrata (bhangriya)

80% ethanolic extraxt of fresh leaves

100, 500, and 1000

500, 335, 250

Emblica ribes (viranga)

80% ethanolic extract of dried seed

100, 500 and 1000

375, 250, 187

Hydrocotyle asiatica (khulkhundi)

Aqueous extract of dried leaves

5, 50, 500, and 1000

-

Holarrhena antidysenterica (kutaja)

80% ethanolic extract of dried stem bark

100, 500, and 1000

500, 335, 250

Moringa pterygosperma (sahjuna)

80% ethanolic extract of fresh root bark

100, 500, and 1000

500, 335, 250

Nyctanthes arbortristis (harsingar)

Aqueous extract of dried leaves

5, 50, 500, and 1000

-

 

50% ethanolic extract of dried leaves

5, 50, 500, and 1000

100, 300, 1000

Ocimum sanctum (tulsi)

80% ethanolic extraxt of fresh leaves

100, 500, and 1000

500, 335, 250

Table 1 (continued)

Plant name in Latin and Hindi or English

Parts and preparation used

Doses used (m g/ml)

in vitro in vivo

Parthenium hysterophorus (congress grass)

Juice of fresh flowers

1:2, 1:20. 1:200

-

 

Aqueous extract of fresh flowers

5, 50, 500, and 1000

-

 

50% ethanolic extract of fresh flowers

5, 50, 500, and 1000

100, 300, 1000

Pongamia glabra (karanj)

80% ethanolic extract of dried seed

100, 500 and 1000

500, 335, 250

Streblus asper (siora)

Aqueous extract of dried leaves

5, 50, 500, and 1000

-

Smilax china

80% ethanolic extract of dried roots

100, 500 and 1000

500, 335, 250

Tinospora cordifolia (giloe)

80% ethanolic extract of fresh stem

-

125, 80, 65

Xanthium strumarium (chhota gokhuru)

Juice of fresh leaves

1:2, 1:20. 1:200

-

 

Aqueous extract of fresh leaves

5, 50, 500, and 1000

-

 

50% ethanolic extract of fresh leaves

5, 50, 500, and 1000

100, 300, 1000

Table 2. Minimum effective dose of medicinal plants tested against trypanosome in vitro and in vivo.

Medicinal plant and part used

Preparation used

Minimum effective dose

in vitro1 in vivo

(µg/ml) (mg/kg)

Xanthium strumarium leaves

Fresh juice

1:20

not effective

 

Aqueous extract

500

not effective

 

Alcoholic extract

5000

> 100

Parthenium hysterophorus flower

Fresh juice

1:20

not effective

 

Aqueous extract

50

not effective

 

Alcoholic extract

5000

> 100

Nyctanthes arbortristis leaves

Aqueous extract

1000

not effective

 

Alcoholic extract

1000

not effective

Aristolochia indica stem

Alcoholic extract

500

not effective

1 All preparations had 100% trypanocidal activity in vitro.


The potential of Tinospora rumphii as an anthelmintic against H. contortus in goats

Tomas. J. Fernandez

Parasitic infections are an important economic problem of the livestock industry in the Philippines. They reduce productivity and cause sterility, abortion, and death of producing animals. In small ruminants, Haemonchus contortus is one of the most pathogenic roundworms.

The life cycle of H. contortus has three phases: (1) The adult worm produces in the animal host eggs, which then develop outside of the host, (2) the infective larvae (= L-3) emerge, and (3) the adult stage of the worms develops in the animal host (Olsen 1974). Any disruption in one of the phases adversely affects the development of H. contortus.

Anthelmintics have been used for the control and treatment of haemonchosis (Dumag and Orinion 1978). However, these are very expensive and often not available in remote areas. Hence, local farmers resort to indigenous plants for deworming their animals.

Consultations with farmers in the neighbouring barangays (villages) of the Visayas State College of Agriculture in Baybay, Leyte, Philippines, revealed 18 plants which the farmers in this region were using to deworm their goats. The efficacy of these plants against worms was studied adopting the farmers' practices. During the initial screening, Tinospora rumphii proved highly effective against H. contortus: When the plant's crude extract was administered to goats at a dosage of 40 ml/kg body weight, its percent efficacy was 85.6% and the number of parasite eggs per gram faeces was significantly reduced (Fernandez 1991).

T. rumphii has many uses. In people, the juice from the crushed stem is considered a diuretic and a cure for colds, anorexia, diarrhoea, and cough. The plant's vegetative parts are used for birth control. An aqueous extract is applied for stomach trouble, indigestion, diarrhoea, topical ulcers, and malaria. Wounds, athlete's foot, and scabies are treated with a decoction of the vine (De Guzman-Ladion 1985). T. rumphii powder prepared with coconut oil is used for rheumatism and flatulence of children (De Padua, Lugod, and Pancho 1977).

In an interview with Filippino farmers in Leyte, Oquias (1990) noted that the farmers pounded or chopped the stem of T. rumphii, mixed it with coconut oil or table salt or both and applied the mixture to wounds of carabaos (buffaloes). They also gave a decoction of the plant's stem to their buffaloes for diarrhoea. However, they were not aware of the anthelmintic effects of this plant.

T. rumphii's efficacy against Haemonchus contortus and its ED50 and LD50 are not documented in the available literature. Therefore this study was undertaken with the following objectives:

  • To determine the effective dose of T. rumphii against H. contortus.
  • To study the ED50 and LD50 of T. rumphii.
  • To assess the therapeutic index of a drug made from T. rumphii.

Materials and methods

The study consisted of two phases: (1) pharmacological study of the plant residue, and (2) a controlled study of the efficacy of the plant drug.

Pharmacological study

Preparation of the plant drug

The stems of the plant were chopped, air-dried at room temperature and boiled for 15 minutes on a hot plate using a 1:2 ratio (w/v, 1 part plant parts/ two parts distilled water). Subsequently the extract was concentrated in the rotavapor. The concentrated extract was placed in an oven at 40°C for overnight and the following day the plant residue was collected and kept in a sterile bottle in the refrigerator for use for one week.

Mean lethal dose (LD50)

This was monitored in strong A albino mice of different sexes and ages, equally distributed in each treatment group, with 10 mice assigned to each treatment. The treatment group corresponded with the dose level of the plant drug, with untreated mice as the control group. The mice were observed daily for one week after dosing them with the appropriate amount of the plant drug. When half of the experimental mice showed signs of toxicity or died, the experiment was terminated. However, when less than half of the experimental animals died or showed signs of toxicity, the dose level was increased by logarithmic method. The LD50 of a particular plant drug was computed using the Probit analysis

Mean effective dose (ED50)

To save resources, the in vitro action of the plant drug against the larvae (L-3) of the parasite was first tested. For this, different dosage levels with increasing amounts were used. The dose level at which all L-3 died after exposure with the plant drug was used as the base line for the in vivo ED50 study in goats.

The distribution of the treatment groups and the number of animals assigned in each treatment group for the ED50 study and the computation of ED50 were more or less the same with the LD50 study.

Efficacy study

Culture of H. contortus larvae (L-3)

Faeces were taken directly from the rectum of naturally infected goats. The culture of L-3 larvae from the eggs of infected faeces and their retrieval followed the procedure described in Technical Bulletin No. 18 (1977).

Grouping of experimental goats

Native goats of varying ages and sexes were infected with 2000 L-3 each. After 20 days, their faeces were examined to find out the patency period and determine the number of worm eggs per gram faeces (e.p.g.). This was done three times within one week. The average e.p.g. for the three faecal egg counts constituted the average pre-treatment e.p.g..

The goats were assigned to three groups (T0, T1, and T2), each consisting of five moderately (80-100 e.p.g.) and five heavily infected animals (e.p.g. > 100) (Fernandez 1993).

T0 served as untreated and T1 as positive control. After the last pre-treatment e.p.g., T1 was treated with a commercial anthelmintic containing mebendazole and T2 with capsules of T. rumphii at a dose of 4.5 g/kg body weight.

One week after treatment, the faeces of all experimental goats were again examined three times within one week to determine the post-treatment e.p.g. Finally, the goats were slaughtered and the adult worms in their intestines counted.

Statistical analysis

The percent efficacy (Reik and Keitz 1954) between treated and control groups and between treated groups was compared with the independent t-test (Sokal and Rohlf 1981). The means of pre- and post-treatment e.p.g. and the number of adult worms counted at necropsy were analysed by F-test. The significance of differences between means were analysed for the least significant difference (LSD).

Results and discussion

Because solid drugs are easier to administer than liquids, the pharmacological and efficacy studies used a solid form of T. rumphii produced through concentrating the plant's extract with a rotavap. However, this way of preparing the plant residue did not in any way alter its chemical composition.

The results of the pharmacological studies showed that the LD50 of the residue of T. rumphii was 7.95 g/kg body weight. At this dosage level, 50% of the experimental albino mice showed difficulties in breathing, uncoordinated movement, leg tremours and/or death within 30 minutes after dosing. On the other hand, the ED50 of the plant drug was 4.5 g/kg body weight. At this dose, 50% of the infected goats significantly reduced their worm burden.

Table 1 shows the mean e.p.g. of experimental goats. Statistical analysis of the number of worm eggs per gram faeces before treatment showed no significant difference (p < 0.05) between the control group (T0) and the treated groups (T1 and T2). However, after treatment the e.p.g. in T1 and T2 was significantly reduced when compared with T0. Comparison of the e.p.g. of T1 and T2 showed no significant difference. This means that the plant residue was as effective as the commercial dewormer in reducing the number of worm eggs in the faeces.

Table 1. Mean number of eggs per gram faeces (e.p.g.) and adult worms recovered at necropsy.

Treatment

Eggs before treatment1

Eggs after treatment1

Adult worms1

Control (T0)

1633.3a

2466.0a

1466.7a

Commercial anthelmintic (T1)

1193.3a

37.5b

84.5b

T. rumphii (T2)

1568.9a

245.0b

235.0b

1 Means marked with the same letter are not significant at 5% level.

The mean number of adult worms recovered at necropsy is also shown in Table 1. The worm burden was significantly reduced in T1 and T2 when compared with T0. However, the statistical difference of the number of adult worms recovered in T1 and T2 was not significant at 5% level. Again, this indicates that the plant drug is as effective as the commercial dewormer.

However, it is not known which compound is responsible for the anthelmintic action of the drug made from T. rumphii and what the pharmacodynamics of this compound are.

Conclusion

The results of this and previous studies showed that the crude extract of T. rumphii given at a dose of 40 ml/kg body weight significantly reduced the number of worm eggs in the faeces of naturally infected goats. The LD50 of the plant drug was 7.95 g/kg body weight and the ED50 4.5 g/kg body weight. Based on the reduction of the number of eggs per gram faeces after treatment and the number of adult worms recovered at necropsy, it can be concluded that the extract of T. rumphii is as active as the commercial dewormer it was compared to. These observations demonstrate the potential of T. rumphii as an anthelmintic.

References

De Guzman-Ladion. 1985. Healing Wonder Herbs. Guide to the Effective Use of Medicinal Plants. Philippine Publishing House, Manila, Philippines.

De Padua, L. S., G. C. Lugod, and J. V. Pancho. 1977. Handbook on Philippine Medicinal Plants. Documentation and Information Section, Office of the Director of Research, University of the Philippines at Los Baños, Philippines.

Dumag, P. U. and G. C. Orinion. 1978. Anthelmintic efficacy of albendazole against gastro-intestinal helminths in naturally infected cattle and carabaos. The Philippine Journal of Animal Industry 33:1-86.

Fernandez, T. J. 1991. Local plants having anthelmintic values. ASEAN Journal on Science and Technology for Development 8(2):114-119.

Fernandez, T. J. 1993. Laboratory Manual in Experimental Parasitology. Visayas State College of Agriculture, Baybay, Leyte, Philippines.

Olsen, W. O. 1974. Parasites, Their Life Cycles and Ecology (3rd edition). University Park Place, Baltimore.

Oquias, L. 1990. Survey of medicinal plants that are commonly used by farmers in Leyte. Undergraduate thesis, Visayas State College of Agriculture, Baybay, Leyte, Philippines.

Reik, T. J. and B. Keitz. 1954. Studies on anthelmintics in cattle. Australian Research Journal 33:162-173.

Sokal, R. R. and J. Rohlf. 1981. Biometry. The Principles and Practices of Statistics in Biological Research (2nd edition). W. H. Freemann & Co., San Francisco, USA.

Technical Bulletin No. 18 (1977). Manual of Veterinary Parasitological Laboratory Techniques. Ministry of Agriculture, Fisheries, and Food. Her Majesty Queen of England, London, UK.


Therapeutic efficacy of a herbal preparation in dermatological conditions of animals and its influence on wound healing

B. P. Manjunatha

Introduction

The skin is the largest organ of animals and covers the whole body. It is frequently exposed to various injurious or infectious agents resulting in dermatitis. Skin diseases in domestic animals are of great economic importance as they severely affect livestock production and health. Some of them are of zoonotic importance too. Diverse etiological factors involved in skin conditions often make diagnosis complicated, due to mixed or superimposed infections. Numerous possible origins of skin lesions generally necessitate empirical therapy with broad-spectrum agents. Before the discovery of penicillin and sulphonamides, many herbs in different forms were used for wound dressing. Nowadays, modern techniques are applied for synthesising herbal preparations and modern dosage forms like ointment and cream can be standardised for particular potency.

Himax™ is a topical herbal ointment containing Sida veronicaefolia, Tagetes erecta, Cedrus deodara, and Pongamia glabra. This paper provides a comprehensive review of various studies on this herbal preparation.

In vitro studies

Himax is a judicious combination of herbal extracts and oils. One of its major ingredients is Cedrus deodara. Its volatile oils contain sesquiterpenes (e.g., himachalol and himadarol) and flavanoids (e.g., cedrin). They were found to be highly effective against gram-positive and gram-negative bacteria (Chopra 1960), mites (Gupta 1968), and various species of dermatophytes (Dikshit and Dixit 1982).

Another major ingredient of Himax is Pongamia glabra. Its oil contains furanoflavanoids (karanjin) and furanodiketones (pongamal) which have been reported effective against a range of pathogenic bacteria (Patel and Trivedi 1962).

Bio-autographic localisation of anti-microbial principles was done by fractionating Himax into different solvent fractions and subsequently separating these through thin-layer chromatography (TLC). The TLC plates were then subjected to the bio-autographic procedure described by Rahalison et.al. (1993). The retention factor values of active spots were recorded (D'Souza 1997). This study is a milestone in herbal research, as it forms the first step in isolation and identification of active principles.

The suitable combination of herbal extracts and oils is the reason behind the wide spectrum of activity to Himax. The spectrum was evaluated further by quantifying the minimum inhibitory concentration (MIC). The in vitro efficacy of the drug against various species of fungi and bacteria was interpreted based on the absence of growth of bacteria or fungi at minimum concentrations of the drug in the agar. Table 1 shows the minimum inhibitory concentration (mg/g) for various pathogenic species of bacteria and fungi. The study confirmed that Himax has a potent bacteriostatic and fungistatic action (D'Souza and Jhon 1996).

Table 1. Minimum inhibitory concentration (mg/g) of Himax against bacteria and fungi.

Organisms

Himax

Staphylococcus aureus

6.25

Pseudomonas aeruginosa

12.50

Proteus vulgaris

6.25

Bacillus subtilis

12.50

Trichophyton mentagrophytes

1.25

Trichophyton rubrum

1.25

Microsporum canis

1.25

Candida albicans

25.00

Studies in domestic animals

The results of the above mentioned in vitro studies against the isolates of common invading bacteria and fungi were confirmed by further evaluations through controlled and field studies in various species of domestic animals (Table 2).

Rai and Sastri (1976) reported the complete cure of fungal disease caused by Trichophyton species in dairy animals when treated with Himax. Similarly, Bali and Singh (1979), Nisal (1978), and Tripathy et.al. (1981) confirmed the anti-fungal activity of Himax in various cases of ringworm infection caused by Trichophyton. Himax was also found to be a quick and very effective miticide. The experimental chemotherapeutic trials against sarcoptic mange revealed that Himax is quite effective in treating mange (Housaine and Ruprah 1979). Also Maskey and Ruprah (1984), Ruprah et al. (1981), Tripathy and Acharjyo (1990), and Tripathy et al. (1989) evaluated Himax for its efficacy against mites. They confirmed that Himax effectively treats mange of domestic animals. Singh and Satija (1989) reported the efficacy of Himax in treating pyoderma predisposed by sarcoptic mange and caused by Staphylococcus species. Misquita and Jagadish (1989) reported the clinical cure of canine pyoderma caused by coagulase-positive Staphylococcus, Pseudomonas, Proteus, and other agents. The study concluded that Himax cures both superficial and deep pyoderma within 14-22 days.

Table 2. Number of cases in the species in which Himax was evaluated.

Species

No. of cases

Cattle

1024

Pigs

282

dogs

230

Goats

171

Horses

45

Sheep

30

Rabbits

10

Camels

7

Elephants

1

Total

1800

Apart from specific infections, Himax was studied as a dressing ointment in the treatment of wounds on different parts of the body, of different origin and in different animal species, e.g., traumatic wounds, lacerated wounds, horn evulsion, yoke gall, rope gall, wounds with maggots, FMD foot lesions, and others (Table 3). The clinical observations of the study have established that Himax is quite effective even in the presence of pus and tissue debris or allergic reactions. Himax removes the devitalised tissue, checks the infection, kills the maggots, and acts as a fly repellent. Microbial resistance to Himax has not been encountered. Himax has also been proved to be safe for the people handling it (Angelo et al. 1974, Joshi 1976, Sharma et al. 1981).

Himax - its influence on wound healing

Healing is a most amazing biological phenomenon, aimed at restoring the tissues to complete normalcy of structure and function. The repair of the damaged tissue is either by regeneration or connective tissue replacement. Though connective tissue replacement is considered to be less effective than regeneration, we need to remind ourselves that the entire inflammation and healing process is survival oriented, and that the other alternative, death, is even less acceptable than is scarring (Slauson and Cooper 1990).

The various histological and histochemical studies conducted on the effect of Himax on wound healing revealed that Himax increased the mitotic activity and vascularity of granulation tissue. There was excessive deposition of granulation tissue, early wound contraction, and epithelisation which enhanced wound healing. Wounds treated with Himax showed high concentrations of trace elements like copper, iron, magnesium, and zinc in granulation tissue at different stages of wound healing. As these trace elements are associated with various enzymes as co-factors and intimately related with growth, repair, and regeneration, the high concentrations of these trace elements indicate an increased rate of metabolism at the site of wound healing resulting in faster wound healing. Himax had a highly significant effect on the production of high quantities of hexosamine, a component of granulation tissue, during early periods of wound healing. Significantly increased levels of hydroxyproline were observed in wounds treated with Himax which indicated improved collagen deposition as hydroxyproline is one of the constituent amino acids of collagen (Chandrapuria et al. 1981, Ghani et al. 1981, Pandey and Ghani 1981, Varshney and Verma 1990). The studies concluded that Himax promotes fast wound healing.

Table 3. Effectiveness of Himax in various skin disorders.

Type of lesion

Total No. of cases

Duration for complete cure (average number of days)

Miscellaneous lesions (broken horn, callosities, bursitis)

374

18.0

Unclassified wounds

300

17.5

FMD lesions

250

6.5

Surgical wounds

228

11.8

Mange

170

18.5

Mycosis

167

16.0

Chronic lesions

73

9.0

Lesions with maggots

65

6.6

Bacterial infections

56

12.0

Infected wounds

41

18.0

Non-specific lesions

26

9.0

Footrot

19

7.0

Eczema

14

17.5

Pox lesion

9

8.0

Contagious ecthyma

8

8.0

Total

1800

12.9

Conclusion

This review demonstrates that Himax ointment has a broad spectrum of actions and it is effective in treating wounds and skin disorders caused by various etiological factors. In addition, Himax by virtue of its properties also promotes fast wound healing.

References

Angelo, S. J., S. S. Mishra, and G. S. Malik. 1974. Clinical observation with HIMAX - a vulnerary agent on large animal practice. U. P. Veterinary Journal (Volumes 2 and 3).

Bali, M. K. and R. P. Singh. 1979. Prevalence of ringworm and its treatment with HIMAX. Indian Veterinary Journal 3:39-40.

Chandrapuria, V. P., S. K. Pande, and G. S. Rathore. 1981. Influence of certain loca1 medicaments on trace elements turn over during wound healing. Indian Journal of Indigenous Medicine (Suppl. 1).

Chopra, C. L. 1960. In-vitro antibacterial activity of oils from Indian medicinal plants I. Journal of the American Pharmaceutical Association for Science Education 49:780-781.

Dikshit, A. and S. N. Dixit. 1982. Cedrus oil - a promising antifugal agent. Indian Perfumer 26(2-4):216-227.

D'Souza, P. 1997. Unpublished data. R&D Center, Natural Remedies Pvt. Ltd., No. 5-B, Veersandra Industrial Area, Bangalore 561229, India.

D'Souza, P. and S. Jhon. 1996. Unpublished data. R&D Center, Natural Remedies Pvt. Ltd., No. 5-B, Veersandra Industrial Area, Bangalore 561229, India.

Ghani, A., S. K. Pandey, and G. N. Kolte. 1981. Histological & histochemcial evaluation of certain local medicaments as accelerator of wound healing. Indian Journal of Indigenous Medicine (Suppl. 1).

Gupta, R. K. 1968. Studies of the curative effect of Cedrus deodara oil against sarcoptic mange in buffalo calves. Indian Journal of Veterinary Science 38(2):203-209.

Housaine, M. N. and N. S. Ruprah. 1979. Treatment of sarcoptic mange in dogs. Thesis, Haryana Agricultural University, India.

Joshi, M. R. 1976. Efficacy of HIMAX in the treatment of wounds. A report of 42 cases. Pashudhan (issue 29-30 Sept.-Oct.).

Maskey, D. K. and N. S. Ruprah. 1984. Studies on life cycle of psoroptic mange in buffaloes and chemotherapy under experimental conditions. Indian Veterinary Journal Sept. 1984:740-743.

Misquita, A. A. R. and S. Jagadish. 1989. C1inica1 efficacy of herbal preparations in pyoderma in dogs. Indian Journal of Indigenous Medicine 6:1-4.

Nisal, M. B. 1978. Use of HIMAX in bovine ringworm. Pashudhan (Issue 47-49).

Pandey, S. K. and A. Ghani. 1981. Clinical evaluation of certain local medicaments as accelerator in wound healing. Indian Journal of Indigenous Medicine 2:1-10.

Patel, R. P. and B. M. Trivedi. 1962. The invitro antibacterial activity of some medicinal oils. Indian Journal of Medicinal Research 50:218.

Rahalison. L., M. Hamburger, M. Moned, E. Frenk, and K. Hostettmann. 1993. Antifungal tests in phytochemical investigation: comparison of bioautographic methods using phytopathogenic and human pathogenic fungi. Plant Medicine 60:41-43.

Rai, H. T. and K. N. V. Sastry. 1976. Efficacy of HIMAX as antifungal agent in dermatomycosis. Indian Veterinary Journal 53:228-229.

Ruprah, N. S., P. K. Varma, and M. B. Chabra. 1981. Evaluation of some acaricide against psoroptic mange in buffaloes. Haryana Agricultural University Journal of Research 10(2):309-313.

Sharma, H. N., K. N. Debe, and S. C. Pathak. 1981. Clinical trial of HIMAX in treatment of wounds in animals. Pashudhan (Issue 64).

Singh, N. and K. C. Satija. 1989. Observations on the treatment of buffalo calves suffering from psoroptic mange and pyoderma. Indian Veterinary Journal 4:4.

Slauson, D. O. and B. J. Cooper. 1990. Mechanism of diseases (2nd ed). Williams & Wilkins, London.

Tripathy, S. B. and L. N. Acharjyo. 1990. Sarcoptic mange in camel and its treatment. Indian Journal of Indigenous Medicine 7:21-26.

Tripathy, S. B., S. C. Dash, N. C. Sahool, and D. Siya. 1981. Treatment of ringworm in crossbred heifers with HIMAX. Indian Veterinary Journal 58(3):239-40.

Tripathy, S. N., S. B. Tripathy, and P. K. Das. 1989. Sarcoptic mange in dogs and its therapy. Indian Journal of Indigenous Medicine 6:41-48.

Varshney, A. C. and M. C. Verma. 1990, Efficacy of HIMAX in wound healing: A clinical study in bovine. Indian Journal of Indigenous Medicine 7(1):31-34.


Zeetress - a promising anti-stress remedy and immuno-modulator. A review

C. B. Pande

Introduction

Intensive management, high animal densities, and the high and fast production of eggs and meat have stimulated a renewed interest in stress in animals. Most of the time stress remains unnoticed but it can cause severe internal damages which negatively affect production. Stress leads to heavy losses in the livestock industry each year.

Anything disrupting the physiological and psychological stability of an organism is a stressor and the reaction of an organism to a stressor is termed ‘stress'. In other words, a stressor can be defined as any factor that challenges the status of an organism and forces the individual to make adjustments as the response.

The following responses to stress have been observed:

  • Hormone release: Increased level of plasma corticosteroid.
  • General response: Altered feed intake, energy usage, and body temperature.
  • Protein metabolism: Increased acute phase protein and muscle degeneration.
  • Mineral metabolism: Increased synthesis of mineral-binding proteins.
  • Vitamins: Increased depletion of Vitamin C (ascorbic acid).

Presently emphasis is directed towards the search for herbal formulations which can be helpful in the management of stress and related disorders. The immuno-modulating effects of several herbal formulations have been studied and a number of clinical and experimental trials have been conducted to evaluate the therapeutic properties of such herbal formulations. Many of the studies have shown very encouraging results.

In this context, several Indian medicinal plants have been reported to possess adaptogenic, anti-stress, and immuno-modulating activity. For example, Withania somnifera, Ocimum sanctum, and Emblica officinalis have been demonstrated to have significant anti-stress and adaptogenic activities (Bhattacharya and Ghosal 1994). Pande and Vijaykumar (1994) and Rao et al. (1996) successfully demonstrated the immuno-modulating activity of a combination the extracts of these plants.

Zeetress™ is one such formulation known to arrest the damaging effects of stress with strong immuno-modulating activity. It is a poly-herbal formulation of quality extracts of W. somnifera, O. sanctum, and E. officinalis. This article reviews the adaptogenic, anti-stress, and immuno-modulating activity of Zeetress.

Adaptogenic activity of Zeetress

Adaptogenic drugs help the organism to cope during stressful situations. Bhattacharya and Ghosal (1994) demonstrated the adaptogenic activity of Zeetress in laboratory animals by forcing mice to swim in a tank of 30x45x40 cm filled with water and kept at room temperature of about 25 ± 2°C. The duration of swimming till the time when a mouse could swim no further and started getting submerged in the water, was recorded. The results are given in Table 1.

Table 1. Swim survival time in mice without and with Zeetress (Bhattacharya and Ghosal 1994).

Groups

Swim survival time (minutes)

Improvement (%)

Control

182 ± 24

-

Zeetress

318 ± 29

75

In the same study, another set of mice was made to swim in the tank in water kept at 20 ± 2°C, until the mice's rectal temperature reached 20 ± 2°C as assessed by a telethermometer with rectal thermistor probes. At this point, the mice were taken out and kept at room temperature of 32 ± 2°C. The time taken for the rectal temperature to fall to 20 ± 2°C and then to recover to 37 ± 1°C at room temperature was recorded. One group received Zeetress while another group remained untreated as control. The results are depicted in Table 2.

Table 2. Effect of cold stress in mice without and with Zeetress (Bhattacharya and Ghosal 1994).

Groups

Time for inducing hypothermia (minutes)

Improvement (%)

Time for returning to normothermia (minutes)

Improvement (%)

Control

52.4 ± 4.6

-

129.6 ± 7.2

-

Zeetress

98.8 ± 7.3

88.54

82.5 ± 5.9

57.1

A further set of mice was subjected to anoxic stress by keeping them in hermetically sealed jars. The time taken for the mouse to exhibit clonic convulsions was the end point and thereafter the animals were immediately removed for recovery. Table 3 shows the results of this trial.

Table 3. Effect of anoxic stress in mice without and with Zeetress (Bhattacharya and Ghosal 1994).

Groups

Time to convulsions (minutes)

Improvement (%)

Control

128.8 ± 12.6

-

Zeetress

168.4 ± 8.2

30.75

In an extensive, large-scale field trial Wheeler (1993) demonstrated the adaptogenic activity of Zeetress in commercial broilers. He noted that birds which had received Zeetress had better body weight and feed conversion and lower mortality than the untreated control group (Table 4).

Table 4. Adaptogenic activity of Zeetress in broilers with and without Zeetrees (Wheeler l993).

Groups

No. of broilers

Average body weight (kg)

Feed conversion rate

Liveability (%)

Control

100 000

2.21

2.22

91.72

Zeetress

100 000

2.56

2.10

95.51

Anti-stress activity of Zeetress

The hypothalamus initiates the responses to a stressor. It induces the anterior pituitary to increase the production of adrenocorticotropic hormone (ACTH) which in turn reaches to the adrenal gland and causes hyperactivity of its cortical cells resulting in an increased synthesis of cortisol, also a hormone. If the increased level of cortisol continues for a long time, it causes marked catabolism of protein to combat the stress which results in poor production, general resistance, and decreased antibody response (Swenson 1970). When the plasma concentration of corticosterone is elevated to levels associated with stress response, the growth rate gets reduced. This reduction is associated with increased protein breakdown (Sharpe et al. 1986).

Bhattacharya and Ghosal (1994) have successfully demonstrated the plasma corticosterone-regulating activity of Zeetress. To induce of stress, they immobilised mice through fixing them on a wooden plank with the help of an adhesive plaster for 2 hours at 4°C. Subsequently the adrenal glands were removed and their corticosterone concentration estimated. Blood was collected and the plasma corticosterone was estimated. The results indicate that Zeetress regularises the plasma corticosterone concentration close to the normal level (i.e., the level measured in the healthy control) in spite of severe stress (Table 5).

Table 5. Stress-induced plasma corticosterone levels in mice without and with Zeetress (Bhattacharya and Ghosal 1994).

Groups

Plasma corticosterone (mcg/dl)

Increase (%)

Unstressed (healthy control)

13.2 ± 1.8

 

Immobilised (stressed control)

29.8 ± 1.9

225.76

Immobilised and treated with Zeetress

17.2 ± 1.4

30.30

Excessive corticosteroid blood levels during stress reduces the number of circulating lymphocytes and increases the number of neutrophils, thus altering the normal neutrophil:lymphocyte ratio (Fraser 1991).

A differential blood count showed that in general, the percentage of lymphocytes decreased and the percentage of heterophils increased following exposure of the hens to a stressor (Wolford and Ringer 1962).

Gowda et al. (1996) also demonstrated an altered ratio of heterophils:lymphocytes by challenging birds with viral-induced stress. They observed that the heterophil:lymphocyte ratio increased several times over the healthy control while in the group that had received Zeetress, the ratio remained very close to the level measured in the healthy control (Table 6). In another field trial, Gauthier (1997) reported similar results.

Table 6. Viral-stress induced heterophil:lymphocyte ratio in birds without and with Zeetress (Gowda et al. l996).

Groups

Heterophil:lymphocyte ratio

Increase (%)

Healthy control

0.495 ± 0.10

-

Viral-induced stress, no Zeetress

2.832 ± 1.37

472

Viral-induced stress plus Zeetress

0.567 ± 0.09

14.55

Ascorbic acid depletion and Zeetress

Under normal circumstances, ascorbic acid (Vit. C) is synthesised in the body (North 1984). During stress severe depletion of Vit. C takes place. Vit. C stimulates the phagocytic activity of leukocytes, the reticuloendothelial system, and the formation of antibodies (Vitamin Compendium, ROCHE).

In an extensive trial Bhattcharya and Ghosal (1994) demonstrated the stress-induced depletion of ascorbic acid and the effect of Zeetress on it in laboratory animals by subjecting them to immobilisation stress. Table 7 shows the results.

Table 7. Depletion of ascorbic acid induced through immobilisation stress in laboratory animals without and with Zeetress (Bhattacharya and Ghosal 1994).

Groups

Ascorbic acid (mcg/100mg)

Depletion (%)

Unstressed (healthy control)

304.2 ± 36.4

-

Immobilised (stressed control)

116.8 ± 16.2

61.60

Immobilised and treated with Zeetress

256.4 ± 22.1

15.71

Gowda et al. (1996) demonstrated a similar effect of Zeetress in commercial broilers by subjecting them to viral-induced stress and observed that the depletion of ascorbic acid was significantly lower (31%) in the group treated with Zeetress than in the negative control.

Table 8. Depletion of plasma ascorbic acid induced through viral stress in broilers without and with Zeetress (Gowda et al. 1996).

Groups

Plasma ascorbic acid (mg/dl)

Depletion (%)

Healthy control

0.708 ± 0.203

-

Viral-induced stress, no Zeetress

0.208 ± 0.022

71

Viral-induced stress plus Zeetress

0.489 ± 0.161

31

Zeetress in cattle

High production, calving, transportation, surgical interventions, and diseases produce severe stress in cattle, resulting in poor performance and severe economic losses.

Singh et al. (1997) demonstrated adaptogenic anti-stress activities of Zeetress in heavy crossbreed cattle during hot summer (107-113°F). Thirteen healthy crossbred animals were selected for the study. Their milk yield, haematological and biochemical parameters were recorded for 15 days prior to the administration of Zeetress. Then a course of Zeetress was administered as per the recommended dose and schedule. The above mentioned parameters were recorded after the treatment and compared with the values recorded before the treatment. It was observed that there was significant improvement in the milk yield (28.87%) and in total serum protein (9.33%) after treatment. The neutrophil:lymphocyte ratio was also reduced (9.23%) after treatment, confirming the adaptogenic anti-stress activity of Zeetress.

Immuno-modulatory effect of Zeetress

Brekhman and Dardymov (1969) opined that adaptogenic agents should not only be tested for their anti-stress property, but also for their immuno-competence. There is evidence that certain adaptogens, particularly medicinal plants, increase the resistance against adverse effects of a number of extraneous factors of physical, chemical, or biological origin.

Ghosal et al. (1989) indicated possible immuno-modulatory effects of sitoindosides IX and X (glycowithanolides) from W. somnifera which is one of the major components of Zeetress.

Pande and Vijaykumar (1994) successfully demonstrated the immuno-modulating effect of Zeetress in commercial broilers by studying the broilers' antibody titres. They observed that the antibody titres against Newcastle disease (ND) significantly improved in the group treated with Zeetress compared to the titres in the control group (Table 9). Also weight gain and feed conversion were better in the treatment group with both groups being kept under the same feeding, management, and health coverage scheme. The results confirmed the stimulation of the humoral immune response through Zeetress.

Table 9. Newcastle disease antibody titres, body weight, and food conversion rate in broilers without and with Zeetress (Pande and Vijaykumar 1994).

Groups

ND antibody titres

Body weight (kg)

Food conversion rate

Control

16, 32, 32, 64, 64, 64, 64, 64, 128, 128

1.42

2.38

Zeetress

32, 64, 128, 128, 128, 256, 256, 512, 512, 512

1.54

2.12

Rao et al. (1996) reconfirmed the immuno-modulatory action of Zeetress in birds and observed that when ND vaccine virus was used as indicator system, there were higher levels of antibody titres in both birds vaccinated against infectious bursal disease (IBD) and unvaccinated birds after the application of Zeetress. There was also a marked increase in rosette forming T-lymphocytes suggesting the stimulation of cell-mediated immunity. This assumption was supported by the significantly delayed and severe type of hypersensitivity reaction that was recorded in the DNCB (2.4-dinitrochlorobenzene) skin sensitivity test. Administering Zeetress had also increased the macrophage activity in the spleen as shown by the increase in the number of formazan-positive cells in the nitroblue tetrazolium test. Zeetress was also responsible for a significantly higher body weight in IBD-vaccinated birds. Histological examination of the bursa indicated that Zeetress can help protect the follicles of the bursa against damage resulting from IBD live vaccine. While in untreated birds the majority of the follicles were atrophied due to the destruction of lymphocytes and intra- and inter-follicular oedema, the follicles of birds treated with Zeetress were partially protected or spared. The outcome of this experiment strongly suggests that Zeetress significantly improves the immune status of IBD-vaccinated chickens.

Chatterjee (1994) also demonstrated that Zeetress potentates both cellular and humoral components of the immune system with consequent increase in the host defence against pathogenic stimuli. Microbicidal activity of polymorph cells was significantly increased in rats treated with Zeetress. Both primary and secondary antibody titres as well as the T-cell-mediated delayed type of hypersensitivity response were positively modulated by this poly-herbal product.

References

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Brekhman, I. I. and I. V. Dardymov. 1969. New substances of plant origin which increase nonspecific resistance. Ann. Rev. Pharmac. 9:419-430.

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