The McNair Scholarly Review 1998. Volume 3: 145-159.

 

Salvia divinorum Epling et Játiva-M. (Labiatae): An Ethnopharmacological Investigation

Sherry A. Rovinsky, Biology

Gerald R. Cizadlo, Ph.D., Department of Biology

ABSTRACT

Salvia divinorum is a vision-inducing and medicinal plant of Mexico. It contains an unidentified acetone-soluble compound(s) which inhibits the growth of rod-shaped bacteria on starch agar. Preliminary testing also indicated that a water-soluble compound(s) in S. divinorum slowed the frequency and increased the duration of phasic contractions in the duodenal smooth muscle of mice.

Introduction

Salvia divinorum Epling et Játiva-M. (Labiatae) (also called Ska Pastora, Ska María Pastora, hierba María, hojas de la pastora, hojas de María Pastora, or yerba de María) is a somewhat rare Mexican plant which has been traditionally used in Mazatec healing ceremonies (1). The divinatory uses of this plant are well documented (1,2,3). While its origin and medicinal applications are not well known, the few available articles suggest some interesting possibilities. Infusions of the plant are used for a variety of complaints: diarrhea, headache, rheumatism, anemia, and panzón de barrego, a magical disease of the Mazatecs (1). The Mazatecs consider it a panacea (1). The purpose of this study is twofold: first, to determine if the plant produces antimicrobial compounds, and second, to create a methodology for measuring its effects on gastrointestinal motility in mice.

Literature Review

Historical Background

Few references to S. divinorum can be found in the literature and those are often vague or incomplete. In 1938, anthropologist J. B. Johnson wrote that an infusion of a plant called "hierba María" produced visions in its users (4). B. P. Reko, a medical doctor with an interest in anthropology and ethnobotany, later collected some of this mysterious leaf, but could not identify it (5). In 1952, anthropologist Weitlaner wrote "Curaciones Mazatecas," in which he described a healing ceremony where S. divinorum was used (6). In 1955, banker R. Gordon Wasson and chemist Albert Hofmann met the Mazatec shaman María Sabina and this single interaction provided most of the original information about S. divinorum (7). During this visit, Wasson and Hofmann obtained a living sample of the plant, which they turned over to botanist Carl Epling (8). Epling and Carlos Játiva later classified it as a distinct species of the Salvia genus (9).

No one knows where this strange member of the Labiatae or Lamiaceae (mint) family originated. Some of the Mazatecs believe that S. divinorum is foreign to their region (10). In his 1963 paper, R. Gordon Wasson suggested that S. divinorum may be pipiltzintzintli (Nauhatl for "the most noble prince" (11)), a magical Aztec plant whose identity is unknown (10). However, Díaz suggests that S. divinorum was actually a post-conquest addition to the Mexican flora and therefore cannot be this ancient Aztec sacrament (3). Indeed, Mazatec names for S. divinorum seem to be rooted in Christian mythology. For example, it is often referred to as Ska María Pastora, or the leaf of Mary the Shepherdess (1) because visions of a woman are a common effect (12). Of course, as Ott points out, Mary is not generally thought of as a "shepherdess" by Christians (13). There are no indigenous names for S. divinorum and Ott explains that the Mazatecs know very little about the use of this plant, citing their mistaken belief that the leaves are no longer psychoactive once dried (3). Instead, Díaz suggests that Pipiltzintzintli is actually Cannabis sativa (14), although today it is generally recognized that Cannabis was a post-conquest addition to the New World. Ott ultimately concluded that wherever S. divinorum came from, it was probably not introduced by the Europeans (3).

According to the Mazatecs, S. divinorum is the most important in a "family" of plants which are all of the Labiatae family. This "family" has great religious significance and includes S. divinorum, Coleus pumila, and two forms of C. blumei. They are referred to by the natives as la hembra (the female), el macho (the male), el nene (the child), and el ahijado (the godson), respectively (15).

Botany of Salvia divinorum

S. divinorum is a perennial herb which grows to a height of 50 to 150 cm. The stems are quadrangular and hollow in cross-section, and the leaves have an opposite arrangement. Today, forest ravine areas in the northeastern Sierra Mazateca mountains in Oaxaca are the only known locations where the plant grows naturally. During the winter of 1984, Reisfield found 15 different populations of S. divinorum in his field work. However, since the plant is easily propagated through cuttings, it is now cultivated in various parts of the United States, preferring sites with indirect light and high humidity. It is assumed to be a hybrid, although its two parent species remain mysteries (16).

The flowers of S. divinorum have been the focus of some past confusion. The flowers have white corollas and purplish calyces (1). In Epling’s 1962 botanical description of S. divinorum, he mistakenly describes it as having blue corollas (9). He had never seen living flowers, and his statement was based on Hofmann’s description of "blue flowers crowned with a white dome" (16) which were actually flowers that had blue calyces and unopened white corollas. The mistake was eventually discovered and corrected by other researchers when the specimen Epling was cultivating bloomed (1).

The viability of seeds produced by S. divinorum is questionable. It is apparently a self-sterile plant which will produce seed only when cross-pollinated (1). Valdés pollinated fourteen flowers and four produced seeds. Unfortunately, the seeds’ viability could not be assessed as they were accidentally killed when a growth chamber overheated (1). Generally, this plant is reproduced by cuttings. It is a diploid species (N=11) whose pollen grains are not as viable as those of other Salvia species. Out of 3027 pollen grains, Reisfield found that 53% aborted, although this fact alone does not explain why the plants do not set seed in Mexico (16). Even when S. divinorum is hand pollinated, only 2 to 3 percent of the nutlets fully mature (8).

Valdés had difficulty with insect infestations within the greenhouse. His plants suffered whitefly and Tetranychus urticae infestations, but when they were taken outside, wind and rain prevented these infestations (1).

Valdés’ botanical experiments indicated that the flowering of S. divinorum is controlled by day length. Long nights (12-13 hours) stimulate bloom formation. Flowering is not dependent on tapering of day length; either a gradual or abrupt decrease (from 16 to 11 hours) will cause the plant to flower (17). Valdés also noted that increasing the day length to greater than 12 hours once flower buds had formed caused the blossoms to abort and vegetative growth to resume (1). Plant height is only a minor factor in flowering (17).

Traditional Mazatec Medicinal Uses

There is little information in the literature about medicinal uses of S. divinorum, but there are many instances throughout the world where other Salvia species are used medicinally (1, 11, 14, 18). In fact, the genus name Salvia itself comes from the Latin word salvare, which means "to save." Customarily, the natives talk about dosages of S. divinorum in pairs of leaves (1). Infusions of the plant are most commonly used (1,3), but it has also been used in water as a poultice (3) and sometimes the patient is bathed in the infusion (3).

Leander Valdés learned about its medicinal uses from the Mazatec shaman Don Alejandro. When taken in small doses (4 or 5 pairs of leaves in a tea) it is useful as a "tonic or panacea." This infusion may be taken by the glass or teaspoonful as needed. Specifically, it is used for the regulation of defecation and urination and reputedly stops diarrhea. It is also used for rheumatism and headache, although higher doses may actually produce headache. Valdés writes, "It is given to the sick, old or dying to revive them or alleviate their illness. People who are pale, white and almost ready to die (they have ‘anemia’) may recuperate on taking la María" (1).

Its most mysterious use is as a cure for a disease called "panzón de barrego." According to the Mazatecs, this disease is caused by the curse of a brujo (sorcerer). The victim’s abdomen swells up like a sheep’s belly (hence the name) due to a "stone" the brujo has placed there. The identity of this affliction is unclear. Taking Salvia eliminates the "stone," and the abdomen shrinks down to its original size. An old shaman showed Valdés his wrinkled abdomen, indicating that he had been afflicted with the "shaman’s curse" and had been cured with S. divinorum. Don Alejandro confirmed the shaman’s story (1).

Valdés participated twice in Salvia ceremonies, ingesting an infusion made of 50 pairs of leaves. He and his party reported sensations of "flying or floating and traveling through ‘space,’ twisting and spinning, heaviness or lightness of the body and ‘soreness,’" (1) dizziness, and lack of coordination (1). The speech of the subjects was slurred and contained "awkward sentence patterns" (1). One man had a decreased heart rate and chills. When light was shined into the subjects’ eyes, pupillary response was normal (1). In a later paper, Valdés described the effects as "astounding visual, oral/aural, and tactile hallucination" (17).

Valdés’ brief description of the disease panzón de barrego (1) suggested that the disease may be a parasitic infection in which lymphatic blockage occurs. Another possibility is that this disease may affect capillary fluid dynamics in such a way that filtration is increased and ascites occurs. Also, its folk use for diarrhea suggested that it might either inhibit bacterial infections or act directly on the smooth muscle of the intestinal tract, thereby decreasing gastrointestinal motility. However, since there is no information on the speed of action and symptoms of panzón de barrego, any conclusion is premature.

Mazatec shamans primarily use S. divinorum as a vision-inducing plant. They say it "allows them to travel to heaven and talk to God and the Saints about divination, diagnosis, and healing" (19). Although the plant is reputed to be only "weakly psychotropic," (1) it can produce very powerful visions under the correct circumstances. Rituals are performed to heighten the experience, and darkness and silence are essential for achieving full psychoactivity. Valdés writes "...if the experience becomes too terrifying, it can be readily terminated by saying a few words or producing a light." Dosages of at least 20 pairs of leaves are necessary for psychoactive effects (1). Although its traditional use is as a hallucinogen, the Mazatec shamans only use it when morning glory seeds and Psilocybe mushrooms are unavailable (20). The Mazatecs generally prepare fresh leaves for an infusion by crushing them in water. This apparently forms a microsuspension or emulsion of salvinorin A (and possibly other non-water-soluble psychotropic agents) (1). The dried leaves cannot be used in this manner, as drying changes the chemical composition of the leaves (19).

There are a variety of other physical symptoms that occur upon ingestion of S. divinorum. Hyperthermia occurs with some subjects (8), and this researcher has spoken to others who experience diaphoresis and chills. According to Wasson, the Mazatec Indians often vomit when they ingest the leaves of Ska Pastora (15), but others report that nausea is very rare and that it is simply the act of swallowing the leaf material that stimulates the gag reflex (8).

Compounds isolated from Salvia divinorum

Salvia divinorum is a source of several pharmacologically interesting compounds. Non-nitrogenous diterpenes (salvinorins A and B) have been isolated from Salvia (1,14). Two separate groups isolated salvinorin A, a furanolactone neoclerodane diterpene, from dry leaves, each independently establishing the structure from X-ray crystallography (21,22). Salvinorin B is the inactive desacetyl derivative of salvinorin A (19). (When Valdés isolated these compounds, he named them divinorins A and B, respectively. However, as the Ortega group preceded Valdés in publication, the proper names for these compounds are salvinorins A and B.) Valdés noticed that there were at least two other terpenoids in his extracts (17). Díaz reported the presence of alkaloids in S. divinorum (14), but the Valdés group was unable to isolate any alkaloids (23). Valdés isolated Loliolide, a natural ant repellent, from S. divinorum (24). Jonathan Ott recently attempted to obtain thujones with steam distillation, but was unsuccessful (3).

Other species in the Salvia genus have been shown to contain antimicrobial compounds. For example, Salvia sclarea was shown to contain various compounds which were active against Staphylococcus aureus, Candida albicans, and Proteus mirabilis (25), suggesting the possibility that S. divinorum might be a source of unique antimicrobials as well.

Pharmacological Investigations

Valdés completed a variety of animal experiments in order to determine the psychoactivity of S. divinorum, particularly salvinorin A. Homo sapiens were excluded as test subjects. Since salvinorin A is insoluble in water, he dissolved it in corn oil and Tween-80, a surfactant. He then added water to form an emulsion which settled easily (23).

He attempted to test his aqueous infusion of extract using the Rat FR4 behavior model. Calculating the human dose to be 0.126 g/kg, he gave the rats oral dosages of 10 times and 32 times the human dose (1.26 g/kg and 4.03 g/kg). There was no disruption of FR4 activity or normal behavior. One of his test animals was killed due to the large volume of extract necessary. Due to financial constraints, he abandoned this part of the experiments (1).

He then used a modified extract of reduced volume to test in the cat limb-flick model. Injection of his crude extraction caused kidney failure (anuria) in his cats, one of which died as a result. The survivors developed sterile abscesses at their injection sites. Valdés speculated that the toxicity of his preparation may have been due to its high tannin content. When he modified the preparation to remove the tannins, he administered a subcutaneous injection of it to two cats, using a dosage of 15.16 mg/kg (100 times the human dosage). He observed that the cats panted and foamed at the mouth, but he did not observe limb-flick, nor any of the other "emergent behaviors" that are sometimes observed in cats under the influence of hallucinogens. He administered another subcutaneous injection to a different test animal, this time using a dosage of 85.75 mg/kg which is 635 times the human dosage. This cat became intoxicated within five minutes, cried, and had a decreased respiratory rate and depth. The cat exhibited nystagmus, lack of coordination, and impaired placing reflex. It still had impaired motor function after 24 hours, but had recovered after 36 hours. His control animals did not exhibit these symptoms. Valdés was limited by the amount of extract he could obtain for these experiments and decided to use mice, which require significantly smaller doses (1).

Valdés used the Horizontal Screen test to determine the level of motor function impairment in mice. His preparation was only slightly active, although the mice appeared to be extremely sedated. Activity in this test seemed to be closely related to toxicity. From these experiments, Valdés calculated the ED50 to be 240 mg/kg and the LD50 at 340 mg/kg (1).

Using Hall’s Open Field test, Valdés administered purified salvinorin A intraperitoneally to mice. He modified the test so that it could be used with mice instead of rats and he increased the time of observation. He recorded squares entered, rearings up on hind legs, and length of immobility. Valdés concluded that his purified salvinorin A was not as active as the TLC fraction that had contained it and suggested that there may be another compound(s) in S. divinorum which would increase or potentiate the effects of salvinorin A. Doses as low as 10 mg/kg decreased all three measures of activity. Salvinorin A had a "sedating" effect on the mice, but salvinorin B had no effect in this bioassay (1).

Although Valdés was able to extract pure salvinorin A from the leaves of S. divinorum, he could not determine with certainty that it is a psychoactive compound (1). Daniel Siebert later isolated salvinorin A in the same manner as Valdés and administered it to 20 human subjects, demonstrating that it is indeed the major psychoactive component. When the leaves or extract were quickly swallowed so that little contact with the oral mucosa was allowed, there were no noticeable effects. However, when the material was kept in contact with the oral mucosa, all of Siebert’s volunteers reported psychoactivity. He suggested that salvinorin A is deactivated by the human gastrointestinal system (2). Ott disagrees, saying that this conclusion is premature, owing to the relatively low dosage utilized by Siebert in his bioassays (3). Siebert also found that the strongest psychoactive effects were produced when pure salvinorin A was vaporized and inhaled by test subjects (2). Valdés ultimately concluded that salvinorin A, the first documented diterpene hallucinogen, was the most potent naturally occurring hallucinogen, effective in dosages as low as 200 to 500 mcg (23).

Siebert describes a variety of psychoactive effects of salvinorin A. He writes, "people report having seen visions of people, objects, and places....out of body experiences are frequent. Occasionally individuals get up and move about with no apparent awareness of their movements or behavior. Some individuals speak gibberish during the most intense phase of the experience, others laugh hysterically." The experience of the subjects varied widely depending on dosage, set, and setting (2).

Based on bioassays, Valdés concluded that salvinorin B was not a psychoactive compound. However, this apparent lack of activity may have been due to the fact that it would be less likely to cross cell membranes because of its lower fat solubility. Also, he suspected that his emulsion was inadequate, saying that salvinorin B may be active if it were vaporized and inhaled by the test subject (19).

NovaScreenTM receptor site screening on salvinorin A (concentration = 10-5 M) showed no significant inhibition for various receptor sites tested (2). The receptor sites investigated are listed in Table 1.

Method

Dried S. divinorum leaves were obtained from Kava Kauaii (Hawaii). To test for antimicrobials, two crude extractions of dried, powdered Salvia divinorum leaf were made. For the first extraction, 100 ml hot distilled water was used to extract 5.016 g dried leaf. Water was heated to 95 degrees Celsius and temperature of the leaf-water mixture maintained for 15 minutes. Filtering yielded 18.5 ml brown fluid. In the second extraction, 4.113 g leaf was extracted at room temperature with 25 ml acetone producing 14.0 ml of a deep, bright green solution. This mixture was diluted with an equal volume of distilled water. The control solution consisted of equal volumes of acetone and distilled water.

These crude extracts were then used in a standard disc-diffusion method (26, 27, 28) against 18 test organisms. Each disc was impregnated with 0.02 mL of the appropriate extract or control and tested against an organism plated on starch agar. The organisms tested were Alcaligenes faecalis, Bacillus brevis, B. cereus, B. polymyxa, B. subtilis, Citrobacter freundi, Enterobacter aerogenes, Escherichia coli, Micrococcus luteus, M. roseus, Proteus mirabilis, P. vulgaris, Pseudomonas aeruginosa, P. fluorescens, Serratia liquefaciens, Staphylococcus aureus, S. epidermidis, and Streptococcus lactis.

To measure the effect of S. divinorum on smooth muscle, 1 inch segments of duodenum were removed from non-fasted common mice (N = 5). Each segment was immersed in Tyrode’s solution and connected to a LafayetteTM 76613 force transducer as shown in figure 1. The force transducer was connected to a LafayetteTM 76406TMG minigraph to record the frequency and strength of contractions. 10.033 g S. divinorum was extracted with 400 ml Tyrode’s solution at 95 degrees Celsius for 15 minutes, yielding 237 ml of a brown solution. Some of this extract was diluted with an equal volume of Tyrode’s solution. The Tyrode’s compartment was warmed continuously in a water bath at 37 degrees C and was alternately flushed with 100% S. divinorum extract, 50% S. divinorum extract, and Tyrode’s solution. Controls were produced by similar extractions of Laurus nobilis. 5.010 g L. nobilis leaves were extracted with 200 ml Tyrode’s solution, yielding 141 ml dull yellow solution.

Findings

The water-soluble components of S. divinorum did not contain antimicrobials. In P. aeruginosa, very slight inhibition of growth of about 1 mm occurred around the edges of the disc. There was no visible inhibition of any of the other test organisms.

The acetone extraction contained components which inhibited growth of certain bacteria. Control discs did not inhibit growth of test organisms. Results of this experiment are given in Table 2.

The antimicrobial effects of the acetone extraction did not depend on gram staining characteristics. A. faecalis and P. fluorescens are gram negative organisms, and the extract had no effect. However, it inhibited the growth of the gram negative organisms C. freundi, E. coli, and P. aeruginosa. Conversely, the solution had no effect on M. luteus, S. aureus, S. epidermidis, and S. lactis, although it did inhibit the growth of another gram positive organism, B. subtilis.

The morphology of the microbe did affect the result. Gram negative and gram positive cocci were in all cases unaffected by the acetone extract. However, gram positive, negative, and variable rods were either inhibited or slightly inhibited. The rods B. cereus, P. fluorescens, and E. aerogenes were the exceptions. Figure 2 shows the division of rod-shaped species into families and their sensitivity to S. divinorum extract.

The effect of S. divinorum on duodenal smooth muscle is not clear, although the extract appears to decrease the frequency of phasic contractions while increasing their duration. Typically, contractions occurred approximately once every two seconds when the muscle was bathed in pure Tyrode’s solution. However, when bathed in 100% S. divinorum extract, contractions appeared to stop completely. When flushed with Tyrode’s solution, the muscle recovered almost completely. When the duodenum was bathed in 50% S. divinorum extract, the muscle would contract for approximately 15 seconds, relax for about 2 seconds, and then contract again. By comparison, when the muscle was bathed in extract of L. nobilis, the muscle would stop contracting and would not recover when flushed with Tyrode’s solution. Figure 3 shows the tracings produced from a single strip of duodenal smooth muscle treated with S. divinorum.

Discussion and Conclusion

Agar diffusion is an excellent method to quickly determine anti-microbial properties, but has a few sources of error. Among them is [1] clerical error in recording data, [2] reader error in measuring zone diameters, and [3] contamination or changes in the bacterial strain being tested (27). Also, agar diffusion may not give an accurate picture of the effectiveness of an antibiotic within a living organism. Microbes may show in vitro sensitivity to an antibiotic, with little or no sensitivity to it in vivo (29).

Our preliminary testing with duodenal smooth muscle produced variable results. In three of the mice tested, 15 to 20 second contractions were recorded, but in two, this result could not be reproduced. This may have been due to problems with the measuring equipment. At times, the muscle was visibly contracting, yet produced no tracing. Also, a larger sample size is necessary for statistical analysis.

 

 

Works Cited

  1. Valdés, L. J. The Pharmacognosy of Salvia divinorum (Epling and Játiva-M): An Investigation of Ska María Pastora (Mexico). University of Michigan, Ann Arbor; 1983.
  2. Siebert, D. J. Salvia divinorum and salvinorin A: new pharmacologic findings. Journal of Ethnopharmacology, 43, 53-56; 1994.
  3. Ott, J. Ethnopharmacognosy and human pharmacology of Salvia divinorum and salvinorin A. Curare, 18(I), 103-129; 1995.
  4. Johnson, J. B. The elements of Mazatec witchcraft. Göteborgs Etnografiska Museum Etnologiska Studier 9, 119-149; 1939.
  5. Reko, B.P. Mitobotánica Zapoteca. Privately published: Tacubaya, Mexico; 1945.
  6. Weitlaner, R. J. Curaciones Mazatecas, Anales el Instituto Nacional de Antropología e Historia 4, 279-285, 1952.
  7. Ott, J. The Age of Entheogens and the Angels’ Dictionary. Kennewick, WA: Natural Products, 1995.
  8. Pendell, D. Plant Powers, Poisons, and Herbcraft. San Francisco: Mercury House, 1995.
  9. Epling, C.; Játiva, C.D. A new species of Salvia from Mexico. Botanical Museum Leaflets Harvard University 20(3), 75-76; 1962.
  10. Wasson, R. G. Notes on the present status of ololiuhqui and the other hallucinogens of Mexico. Botanical Museum Leaflets Harvard University (20)6, 161-193; 1963.
  11. Heffern, R. Secrets of the Mind-Altering Plants of Mexico. New York: Pyramid Books, 1975.
  12. Turner, D. M. Salvinorin: The Psychedelic Essence of Salvia divinorum. San Francisco: Panther Press, 1996.
  13. Ott, J. Pharmacotheon: Entheogenic Drugs, Their Plant Sources and History. Kennewick, WA: Natural Products, 1996.
  14. Díaz, J. L. Ethnopharmacology and taxonomy of Mexican psychodysleptic plants. Journal of Psychedelic Drugs II (1-2), 71-101; 1979.
  15. Wasson, R.G. A new Mexican psychotropic drug from the mint family. Botanical Museum Leaflets Harvard University, 77-84; 1962.
  16. Reisfield, A. S. The botany of Salvia divinorum (Labiatae). SIDA 15(3), 349-366; 1993.
  17. Valdés, L. J.; Hatfield, G. M.; Koreeda, M.; Paul, A. G. Studies of Salvia divinorum (Lamiaceae), an halluciongenic mint from the Sierra Mazateca in Oaxaca, central Mexico. Economic Botany, 41(2), 283-291; 1987.
  18. Duke, J. A. Ethnobotanical observations on the Cuna Indians. Economic botany, 29(3), 278-293; 1975.
  19. Valdés, L. J. Salvia divinorum and the unique diterpene hallucinogen, salvinorin (divinorin) A. Journal of Psychoactive Drugs, 26(3), 277-283; 1994.
  20. Schultes, R. E. Hallucinogenic Plants. New York: Golden Press, 1976.
  21. Ortega, A.; Blount, J. F.; Manchand, P. S. Salvinorin, a new trans-neoclerodanediterpene from Salvia divinorum (Labiatae). Journal of the Chemical Society. 2505-2508; 1982
  22. Koreeda, M.; Brown, L.; Valdés, L. J. The absolute stereochemistry of salvinorins. Chemistry Letters. 2015-2018; 1990.
  23. Valdés, L. J. Divinorin A, a psychotropic terpenoid, and divinorin B from the hallucinogenic Mexican mint Salvia divinorum. Journal of Organic Chemistry, 49(24), 4716-4720; 1984.
  24. Valdés, L. J. Loliolide from Salvia divinorum. Journal of Natural Products. 171; 1986.
  25. Ulubelen, A.; Topcu, G.; Eris, C.; Sönmez, U.; Kartal, M.; Kurucu, S.; Bozok-Johansson, C. Terpenoids from Salvia sclarea. Phytochemistry, 36 (4), 971-974; 1994.
  26. Klein, R. M.; Klein, D. T. Research Methods in Plant Science. Garden City, New York: The Natural History Press, 1970.
  27. National Commitee on Clinical Laboratory Standards. Performance Standards for Antimicrobial Disk Sensitivity Tests, 4th ed. (10)7 Approved standard M2-A4; 1991.
  28. Sleigh, J. D.; Timburg, M. C. Notes on Medical Bacteriology. Edinburgh: Churchill Livingstone, 1981.
  29. Ravel, R. Clinical Laboratory Medicine: Clinical Application of Laboratory Data. St. Louis: Mosby, 1995.

 

 

 

 

Table 1: Receptor sites tested for salvinorin A inhibition

Category

Sites Tested

Sites Tested

Sites Tested

Neurotransmitters

adenosine

alpha 1

alpha 2

beta

dopamine 1

dopamine 2

GABAA

GABAB

serotonin 1

serotonin 2

Muscainic 3

NMDA

Kainate

Quisqualate

Glycine (stry sens.)

Regulatory sites

Benzodiazepine(centrl)

glycine (stry insens.)

PCP

MK-801

Brain/gut peptides

angiotensin Ty2

argvasopressin V1

bombesin

CCK peripheral

substance P

substance K

NPY

Neurotensin

Somatostatin

VIP

Growth factors and peptides

AN1

EG

NGF

Ion channels

calcium (type N)

calcium (type T and L)

Chloride

Potassium (low conduct).

Second messengers

forskolin

phorbol ester

Inositol triphosphate

Monoamine oxidase inhibition

monoamine oxidase A

monoamine oxidase B

 

Table 1: Receptor sites tested for salvinorin A inhibition (2).

 

 

 

 

Figure 1: The muscle testing apparatus used to measure duodenal smooth muscle motility.

 

 

 

Table 2: Zone of Inhibition Diameters for Acetone Extraction

Organism

Average size of zone (mm)

Standard deviation: s (mm)

Zone diameter range* (mm)

Alcaligenes faecalis

0.0

NA

NA

Bacillus brevis

9.6

0.52

1.7

Bacillus cereus

0.0

NA

NA

Bacillus polymyxa

9.7

0.45

1.7

Bacillus subtilis

10.0

0.57

1.9

Citrobacter freundi

9.4

0.61

2.2

Enterobacter aerogenes

0.0

NA

NA

Escherichia coli

8.9

0.53

1.6

Micrococcus luteus

0.0

NA

NA

Micrococcus roseus

0.0

NA

NA

Proteus mirabilis

Slight inhibition**

NA

NA

Proteus vulgaris

Slight inhibition**

NA

NA

Pseudomonas aeruginosa

9.3

0.49

1.3

Pseudomonas fluorescens

0.0

NA

NA

Serratia liquefaciens

9.6

0.33

1.0

Staphylococcus aureus

0.0

NA

NA

Staphylococcus epidermidis

0.0

NA

NA

Streptococcus lactis

0.0

NA

NA

Table 2: Zones of inhibition (N=10)

*Zone diameter range is defined as the difference between the largest and smallest zone diameters.

**Slight inhibition means that the zone of inhibition was cloudy, irregular, and unmeasurable.

***Control discs for all organisms showed no inhibition.

 

 

Figure 2: Bacterial species which were inhibited are shown in normal type. Those that were not inhibited are shown in bold type and those which were slightly inhibited are shown in bold italics.

 

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Figure 3: Tracing (a) shows the contractions of the duodenal muscle before treatment with S. divinorum. In (b), the Tyrode’s solution has been replaced with 100% S. divinorum extract, prepared as described in methods. Tracing (c) shows the partial recovery of the muscle after the chamber has been flushed with Tyrodes. In (d), 50% S. divinorum extract replaced the Tyrodes solution. One division = 0.1 g tension. Paper speed is 5 mm/sec.