Emanations Volume 4

ABSTRACT

Traditional medicine throughout Amazonia is rooted in the belief that imbalances in the spirit world can be the origin of illness. The ingestion of Ayahuasca, a hallucinogenic brew made primarily from the bark of Banisteriopsis sp., along with other “accessory plants,” is believed to establish a connection with the spirit world, which facilitates the diagnosis and cure of various ailments. Through both structured and unstructured interviews, ethnobotanical data was collected on the cultural use of ayahuasca. In the Yarapa River region of Loreto, Peru, three species of Banisteriopsis (Malpighiaceae) can be found: B. muricata, B. martiana, and B. caapi. Bark samples were collected and extracted in a solution of 70% ethanol, 30% water and a minute amount of HCl. A comparative study of the three species was performed using thin-layer chromatography (TLC). Disk diffusion assays were performed on Bacillus subtilus (gram-positive bacteria), Escherichia coli (gram-negative bacteria), and the yeast Saccharomyces cerevisiae to test for biological activity of the prepared Ayahuasca mixture. Although preliminary analysis indicates the presence of alkaloids in all three species, B. muricata and B. caapi exhibit greater biochemical activity than B. martiana.

In the Amazonian world view, there is an intrinsic connection between mental, physical, and spiritual well being. The word Ayahuasca is Quechua for ‘spirit vine’ or ‘vine of the dead.’ It refers both to the woody liana Banisteriopsis sp., and to the brew prepared from it. To the indigenous people of the Amazon River Basin, Ayahuasca is a sacred vine, highly respected for the power it is believed to embody.

Ayahuasca has a long history of use by the indigenous peoples throughout the Amazon basin (Schultes 1957, Creedon 2001). The main component of the Ayahuasca mixture is Banisteriopsis sp. In the Yarapa River region of Amazonian Peru, three species of Banisteriopsis can be found: B. muricata and B. martiana in the flooded forest, and B. caapi in the terra firme forest. Through informal interviews it was found that although B. caapi is the most widely used species in Ayahuasca preparation (as well as the most widely studied scientifically), the other species are sometimes used as the primary plant in the mixture. Studies conducted on B. caapi have detected a number of alkaloids in the bark including harmine, beta-carbolines, harmaline, and tetrahydroharmine (McKenna et al 1984, Rivier and Lindgren 1972). Beta-carbolines function as monoamine oxidase (MAO) inhibitors causing increased levels of serotonin in the brain: preliminary evidence that Ayahuasca may be useful in psychiatry. The basis for the psychoactivity of the prepared ayahuasca mixture is the interaction between the beta-carbolines present in Banisteriopsis and Dimethyltryptamine (DMT) present in Psychotria viridis, an “accessory plant” (McKenna et al 1984). It is the intent of this study to further investigate the ethnomedical uses and biochemical properties of Banisteriopsis sp.

MATERIALS AND METHODS
Collection and extraction

Specimens of B. muricata, B. martiana, and B. caapi were collected from the flooded forest region surrounding the Yarapa River Lodge, neighboring Jaldar Village, and from a region of terra firme forest further upstream. The bark was shredded and allowed to extract for 24 hours in a solution of 70% ethanol, 30% water and a minute amount of HCl. The plant matter was then filtered and the resulting extract was concentrated using a heat-based evaporation method.

Ethnobotanical Data

Ethnobotanical information was collected through both structured and unstructured interviews. Ayahuasca is well known among the villagers and indigenous people; however, its actual consumption is apparently not prevalent in the Yarapa River region. The majority of the ethnobotanical data was obtained from Esteban Mosquera and Ramón Arawanza, two native shaman of the region.

Thin-Layer Chromatography

A comparative study of the three extracts was performed using thin-layer chromatography. Prepared aluminum-backed plates coated with silica gel were used. A solvent solution of 100 parts water to 100 parts ethyl acetate to 200 parts methanol to 6 parts ammonia was used in all three TLCs. The separated compounds were then visualized under UV light, and with an ammonia reagent to detect the presence of organic compounds.

Bioassays

Anti-microbial disk diffusion assays against Bacillus subtilus (gram-positive bacteria) and Escherichia coli (gram-negative bacteria) were performed using a sample of the prepared Ayahuasca mixture obtained from the medicinal plant market in Iquitos. Disks were impregnated with 20uL of the extract, dried, and placed on a Petri plate inoculated with bacterial culture. The plates were incubated for 24 hours at 37 C. In addition, an anti-fungal assay against Saccharomyces cerevisiae was also performed using the same extract and methods.

RESULTS
Comparative Chemistry

TLC analysis detected the presence of alkaloids in all three bark extracts. B. caapi revealed four compounds (pale blue fluorescence at Rf=0.16, medium blue at Rf=0.34, bright green at Rf=0.42, and orange at Rf=0.45). B. muricata revealed four compounds as well (purple fluorescence at Rf=0.07, pale green at Rf=0.20, bright green at Rf=0.42, and blue at Rf=0.50). TLC analysis of B. martiana revealed only one compound (purple fluorescence at Rf=0.7). Comparative analysis found one compound in common between B. muricata and B. caapi (bright green fluorescence at Rf=0.42), as well as a shared compound between B. martiana and B. muricata (purple fluorescence at Rf=0.7).

Bioactivity

After 24 hours, the prepared Ayahuasca mixture was observed to inhibit the growth of B. subtilis. After 36 hours, the zone of inhibition quadrupled. The extract also proved to be active in inhibiting the growth of E. coli and S. cerevisiae.

Ethnobotany

Preparation of the Ayahuasca mixture generally involves boiling Banisteriopsis sp. bark in water, along with the bark of Psychotria viridis and other accessory plants. Through informal interviews it was found that B. caapi is the most widely used species in Ayahuasca preparation. The brew is allowed to boil until most of the water evaporates and only a thickly concentrated brown liquid remains. This usually takes approximately eight hours and is generally done on the day of the Ayahuasca ceremony. The ceremony is always held at night and led by an experienced Ayahuasquero, or shaman specialized in the preparation and administration of Ayahuasca. He is responsible for making sure the participants follow a special diet for at least one week prior to the ceremony, eliminating meat, fish, chicken, and excess sugar and salt.

At the start of the ceremony, the shaman prepares for communication with the spirits by the ritual smoking of Mapacho, a tobacco mixture. The smoking of these freshly rolled cigarettes is often a ceremony in itself as it too is considered to be sacred. The shaman blows smoke onto each participant as a means of asking the spirits to bestow benevolence upon them throughout the ceremony. After this initial phase of the ceremony, each person, including the shaman drinks two and a half ounces of the pungent Ayahuasca brew. After about half an hour, an intense change in perception is said to occur, along with powerful and profound visions, often involving family members, jungle animals and plant spirits. Drinking Ayahuasca induces vomiting, which is considered to be an essential part of the physical, mental and spiritual cleansing process. The hallucinatory effects last from two to eight hours, often followed by another one to eight hours of lingering after-effects.

Ayahuasca is used in the indigenous context for a number of purposes. Rather than a remedy for specific illnesses, it is viewed as a sort of “universal medicine” that allows the shaman to diagnose and cure a wide range of physical and mental ailments through communication in the spiritual dimension. It is believed to “set the spirit right.” This is an important measure against sickness since the spiritual world, rather than the physical, is viewed as a major source of sickness and injury. Ayahuasca is also taken to gain insight into the future, to communicate with dead relatives, and to consult with and obtain knowledge from the plant spirits.

DISCUSSION
Bioactivity and Chemical Analysis

It is interesting to note that one of the less widely studied species of Banisteriopsis, B. muricata was found to contain an equal number of active chemical constituents to B. caapi. The two lesser studied Banisteriopsis species, B. muricata and B. martiana share a compound that is not present in B. caapi and may warrant further study. The prepared Ayahuasca mixture exhibited anti-bacterial and anti-fungal properties. This may be of great consequence to the physical level of the cleansing Ayahuasca provides as a purgative, tying into the complete “mind, body, and spirit” cleansing process.

Ethnobotanical Study

Because shamans continue to use Ayahuasca as a means of spiritual and physical cleansing, the prepared Ayahuasca mixture could potentially be useful in the treatment of certain medical problems in western society. Research has already proven Ayahuasca effective in the treatment of alcohol addiction and potentially useful in the treatment of various other psychological disorders (Grob, et al 1996). The complex interplay of chemical constituents, when combined with the subjective psychological effects of Ayahuasca, may facilitate the patient’s awareness of, and access to, the subconscious workings of the brain (Hoffman, et al 2001). Both the ethnobotanical and biochemical data collected suggest that Ayahuasca has the potential to be a powerful tool in the treatment of various psychological disorders and neurosis. Further study is strongly recommended.

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