MUIH Microbial Analysis Experiment Internship Part III

At last, we have reached the end of this series on the “internship” that is no longer an internship. If you didn’t read the series, you should start at Part I. The point of this blog series is to illustrate some of the work that I have done and things that I have learned. Since this is a real, ongoing, study involving accredited universities (University of Maryland and Maryland University of Integrative Health), I am not disclosing all materials and methods, analysis, or results.

This last portion of the experiment I have been participated in involved testing botanical extracts against various bacteria, yeast, and mold. Specifically, I got to test hydro-alcoholic extracts of Echinacea purpurea (commonly known as Purple Coneflower or Echinacea), of varying strengths to see if it inhibited growth of the bacterias, yeast, and mold. To do this, I created dilutions using 3 different Echinacea tinctures to inoculate, incubate, and interpret results from 3M Petrifilms.

I learned aseptic lab techniques while carrying out the study discussed in Part II of this series. This time around, it was even more important because I was handling petrifilm loaded with yeast and mold spores, and one with E.Coli (yuck!). Having the experience gained from the first round made it much easier to confidently carry out the steps while limiting exposure. Some of the additional daily tasks involved taking ambient temperatures, monitoring samples, counting colonies, and reporting results.

I won’t go into all the nerdy details since more will be written (and it’s really the school’s place to properly publish the study and get credit for it). For now here are some pics of what I’ve been learning and doing.

Rosemary Monograph

Rosemary

img_20180714_081450

Nomenclature

Rosmarinus officinalis

Family

Lamiaceae (formerly Labiatae)

Parts Used

Leaves, flowers, and essential oil.

Identification of Genus/Species

Part	Identification
Stem	Shrub with scaly branches and bark. Grows to 3 feet.
Leaves	1 to 2 inch long needle-like evergreen leaves.
Flowers	Flowers are small and pale blue or white.
Taste	Strong, oily, bitter.
Odor	Strong and pleasant. Camphor-like.

Cultivation

As a Mediterranean native, Rosemary does best in warm, sunny, and dry spots. It can be propagated by seed, cutting, or layering.

Collection

To get the most essential oil, harvest the upper parts before they flower. The flowers and upper parts can be harvested in early Spring and Summer. The leaves and flowers should be dried in the shade.

Constituents

The leaves and flowers contain an essential oil made up of borneol, camphor, 1,8 cineole, linalool, terpenes, and borneol esther.

The leaves also contain tannins, resin, carboxylic acid, and minerals such: as calcium, magnesium, phosphorus, iron, sodium, and potassium.

Actions

Antibacterial, anti-inflammatory, antinociceptive, antispasmodic, antiseptic, astringent, carminative, diaphoretic, emmenagogue, expectorant, hepatic, hypertensive, nervine sedative, rubefacient, stimulant, and tonic.

Medicinal Use

The essential oil is thought to be responsible for the majority of the Rosemary’s therapeutic actions. Rosemary has many uses for the hair and skin: hair growth, dandruff, ulcers, sores and wounds.

Rosemary is used to support colds, headaches, fevers, poor memory, rheumatism, and sprains. Due to its antispasmodic properties, Rosemary is useful for cramps and spasms. It has a general tonic effect on the circulatory system and may be helpful with varicose veins.

Contraindications & Side Effects

Rosemary contains two toxic constituents, borneol camphor and pinene. Camphor in high doses can aggravate asthma and epilepsy.

Rosemary should be avoided during the first trimester of pregnancy. Rosemary should not be administered to children under the age of four.

Rosemary should be tested via a skin patch test prior to topical application.

Follow dosage recommendations for the essential oil carefully. Rosemary can impact blood pressure.

Dosage

Doses can be taken three to four times a day.

Infusion: 3 to 5 tablespoons

Tincture: 5 to 20 drops

Essential Oil: ½ to 3 drops

References

Balick, M. J. (2014). 21st century herbal: A practical guide for healthy living using nature’s most powerful plants. V. Mattern (Ed.). New York: Rodale, 341-345.

Bone, K., & Mills, S. (2013). Principles and practice of phytotherapy modern herbal medicine. Edinburgh: Churchill Livingstone, Elsevier.

Easley,T. and Horne,S. (2016). The modern herbal dispensatory: A medicine-making guide. Berkeley, CA. North Atlantic Books

Hoffmann, D. (2003). Medical herbalism: the science and practice of herbal medicine. Rochester, VT: Healing Arts Press.

Petersen, D. (2015). Herb 201 Herbal Studies. Portland, OR: American College of Healthcare Sciences

Weiss, R., & Fintelmann, V. (2000). Herbal Medicine (2nd ed.). Stuttgart: Thieme

Lemon Balm Monograph

Lemon Balm

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Nomenclature

Melissa officinalis

Family

Lamiaceae (formerly Labiatae)

Parts Used

Leaves

Identification of Genus/Species

Part	Identification
Stem	Similar to others in the Lamiaceae family, the stem is square.
Leaves	Leaves are slightly hairy, broad and ovate. Low lying leaves may be heart shaped. They omit a lemony aroma.
Flowers	Flowers bloom in summer and are small and hooded, in white or lemon color.
Taste	Pleasant, lemony, and mildly spicy.
Odor	Lemony

Cultivation

Lemon Balm is an easy to grow perennial herb that will reach up to three feet high and two feet wide. It is hardy to zone 3 and prefers fertile soil with a slightly acidic pH. Supplementing the soil with different nutrients will impact the chemical composition and essential oil yield.

Lemon Balm can be propagated by seed, cuttings, or division. Lemon Balm grows best with regular watering. It will grow in sun or partial shade.

Collection

The entire above ground herb is harvested. Lemon balm should be harvested by hand in the early morning, after the dew dries. The leaves should be dried immediately.

Constituents

Essential oil contains citronellal, citral, linalool, and other monoterpenes and sesquiterpenes. Also contains tannins, flavonoids, and bitters.

The herb contains caffeic and rosmarinic acids.

The chemical composition of the oil is similar to the pheromone that helps worker bees locate their colonies.

Actions

Antidepressant, antiseptic, antispasmodic, antiviral, carminative, diaphoretic, and nervine sedative.

Medicinal Use

Due to its mild sedative action, Lemon Balm is useful for concentration, depression, sleep and stress. Lemon Balm is also indicated for gastrointestinal disorders and nervous disorders and is especially prescribed for children with these conditions.

The essential oil has antiviral properties so it is used for cold sores and shingles.

Contraindications & Side effects

No known safety concerns.

Dosage

One to three times daily.

Infusion: 1 cup

Liquid Extract 2 to 4 millimeters

Tincture: 2 to 6 millimeters

References

Balick, M. J. (2014). 21st century herbal: A practical guide for healthy living using nature’s most powerful plants. V. Mattern (Ed.). New York: Rodale, 341-345.

Bone, K., & Mills, S. (2013). Principles and practice of phytotherapy modern herbal medicine. Edinburgh: Churchill Livingstone, Elsevier.

Easley,T. and Horne,S. (2016). The modern herbal dispensatory: A medicine-making guide. Berkeley, CA. North Atlantic Books

Hoffmann, D. (2003). Medical herbalism: the science and practice of herbal medicine. Rochester, VT: Healing Arts Press.

Petersen, D. (2015). Herb 201 Herbal Studies. Portland, OR: American College of Healthcare Sciences

Weiss, R., & Fintelmann, V. (2000). Herbal Medicine (2nd ed.). Stuttgart: Thieme

Basil Monograph

Basil

Nomenclature

Ocimum basilicum

Family

Lamiaceae (formerly Labiatae)

Parts Used

Leaves, flowers, and essential oil.

Identification of Genus/Species

Part	Identification
Stem	Grows 3 feet high. Obtusely quadrangular.
Leaves	Leaves grow opposite, are 2 to 3 inches long, oval, and bright green.
Flowers	Flowers are white or pink whorls
Taste	Pleasant, strong, and peppery
Odor	Highly fragrant

Cultivation

Sweet basil, ocimum basilicum, is an annual herb that is very easy to grow. It prefers light, well-drained soil in warm climates with full sun. It will also grow in a container. The top shoots should be clipped to promote fuller, healthier growth. Cuttings can be rooted in water and grown in pots indoors.

Collection

Optimal harvest is just before the blooms open.

Constituents

The exact chemical profile of basil depends on the particular cultivar. Sweet basil contains many chemical compounds in the essential oil: estragole, methyl cinnamate, ocimene, cineole, linalool, thymol, and camphor. The leaves contain tannins, vitamins, and minerals, such as: calcium, iron, potassium, magnesium, B2, Vitamin A, and Vitamin C.

Actions

Analgesic, antibacterial, antiflatulent, antifungal, antiemetic, anticmicrobial, antioxidant, antispasmodic, antivenom, antiviral anxiolytic, circulatory stimulant, digestive, diuretic, galactogogue, hepatoprotective, hypoglycemic, insecticide, kidney tonic, nervine, orexigenic, sedative.

Medicinal Use

Basil has both topical and internal use. Taken orally, basil helps with digestive issues, stomach spasms, kidney issues, and blood sugar issues. Multiple Ocimum species have exhibited a hypoglycemic effect. Basil may also help with headaches, appetite stimulation, circulation, and fevers.

Topically, basil can be used for bites, stings, and may be helpful for acne. Basil can be used as an astringent mouthwash.

Basil essential oil exhibits antibacterial, antiviral, and antifungal properties, making it useful for cleaning and disinfecting.

Contraindications & Side Effects

Basil essential oil is not safe while pregnant, breastfeeding, or lactating. Sweet basil should not be consumed in doses higher than a culinary dose while pregnant, breastfeeding, or lactating and should be avoided entirely by infants and toddlers.

Basil should be avoided by those with an allergy to the Lamiaceae/Labiate families.

There are very few adverse effects reported. Those with allergies to the Lamiaceae/Labiate families could have an allergic reaction to basil.

Dosage

Infusion: 1 cup of tea

Essential Oil: 1 drop

Tincture: 4 millimeters

Fluid Extract: 2 millimeters

MUIH Microbial Analysis Experiment Internship Part II

The second part of this post covers the first phase of the actual experiment. If you missed the first part of this post, you can find it here. Technically, this is no longer an internship because I didn’t do it for college or work credit. After all the research and planning for the experiment, I wanted to be part of it. I was excited about the prospect of participating in a study between two universities, with the possibility of the findings being published.

We were testing the theory that a bunsen burner could create a sterile field. The reason this would be cool is because many product makers are small businesses that cannot afford to rent lab space for product creation. The execution of the experiment required learning and using aseptic lab techniques (limiting variables), meticulously documenting each step and then counting colonies.

IMG_20180607_223610 — The bunsen burner was turned on for five minutes to create the sterile field. Measurements on the table indicate the distance of each TSA plate (a.k.a. a petri dish to most) to the burner. After 5 minutes the lids were removed to expose them to the microbes in the air.

IMG_20180610_015658 — After the predetermined amount of time the lids were replaced and the dishes were wrapped in lab film to keep them secure.

IMG_20180610_015702 — Close up of the dishes before the incubation period.

IMG_20180610_220701 — Here is a couple of days in.

IMG_20180613_085532 — This is after 7 days of incubation. Look at all of those organisms!

The study has not been published yet so I will not discuss the results. Needless to say, my personal results were… gross. It was really awesome learning about aseptic lab techniques and getting to participate in this study.

Part III of this post will be the final part and will cover the second, and last, experiment conducted as part of this study.

HRB 635b Field Trip/ Industry Intensive

HRB 635b is a field trip offered by Maryland University of Integrative Health (MUIH) for students in the Product Design tract of the Masters of Therapeutic Herbalism program. For one week the students travel to Asheville, North Carolina, located in the Appalachian Mountains in the western part of the state. The students have the opportunity to visit various players in the herbal industry, getting behind the scenes tours and hands-on experience. The week is designed in a way that students get to see all aspects of the industry: growing the crops, natural product development, herbal schools, scientific research, conservation efforts, etc.

The purpose of this artifact (and all artifacts on this blog) is to illustrate to the MUIH faculty that I successfully accomplished the objectives of the course. The trip is jam packed with tours, both eye opening and life changing, this may well read like a travel brochure or advertisement for these companies, rather than an educational artifact. I could not even scratch the surface on the work these companies are doing so i encourage interested parties to check out the company pages. Also, feel free to contact me with any questions.

The trip that I participated in was September 5- 10, 2017. The learning objectives for this course were for the student to:

Have developed awareness of diverse business models employed by herbalists in the herbal products industry.
Have an introductory knowledge of the major components of an herbal products company including but not limited to cultivation, harvesting, processing, formulation, quality control, research and business (marketing, sales, and financial considerations)
Have gained an appreciation for the range of sizes/scales of successful herbal products businesses
Have gained a foundational knowledge of the principles of and ethical considerations surrounding sustainable sourcing (cultivated vs wildcrafted, etc.) of medicinal plants for commercial use
Have developed an awareness of the importance of preserving natural plant communities and the pros and cons of sustainable wild harvest of medicinal plants
Have continued to improve their plant identification skills, building on instruction from previous coursework.

In order to achieve these objectives, we visited local businesses and schools, participated in class lectures at the house where we all stayed, did research and homework assignments, and spent some time hiking and wildcrafting. Like many businesses, no cameras were allowed in most operational areas. I have included the photos that I was allowed to take.

Blue Ridge Food Ventures – This facility is a large warehouse, composed of multiple kitchens, that are used to manufacture food, beverages, and natural products. We were able to tour the facility and see how other small businesses owners rented certified kitchen spaces in order to meet Good Manufacturing Processes and regulatory requirements of their industry. This was particularly helpful for those of us that were struggling with the logistics of maintaining a legally complaint business. The photos below were taken inside the front of the facility and showcase some of the products that are manufactured within the building.

Blue Ridge School of Herbal Medicine Blue Ridge School of Herbal Medicine Director (founder), CoryPine Shane allowed us entry into his school.

We all took seats in his classroom as he shared his story. He held nothing back, telling us both the good and the bad that he experienced while building a career in a tough path. He offered wisdom and insight, answered questions, and let us all take a sneak peak into his apothecary, where Pine Herbals are crafted. Not only did I leave with a better understanding of the business side of the industry, but I purchased a pretty awesome bottle of Betony tincture, as well.

Gaia Herbs

Gaia Herbs is spoken, almost in reverence, by many in the herbal industry. All I knew of the company was that they produced high quality herbal supplements – and that I liked their tea. Given the sheet scope and size of their operation, there was a lot to see and discuss.

We started our tour in the chemistry labs where we got to see millions of dollars in cutting edge equipment at work. No photos are allowed within the labs so you are stuck with this one of me in the lobby prior to entry. We learned about the extensive testing that Gaia conducts and got to ask questions of the Quality Assurance team at work. Our discussions of solvents, extraction methods, aseptic lab techniques, etc was easily an hour or two of our day. I have never been in such a nice, well equipped lab, so it was a bit mind blowing to think that it was a lab for a company in the herbal industry. This is when I first started to understand why herbalists root for Gaia.

Next we toured the production facilities. Since our phones were outside in the lobby, I did not get to take extension notes. We had a very good discussion of the various methods for creating herbal extractions. Gaia uses a percolation and their testing has show it to be more effective than other means of extraction. We got to see the machinery involved and ask questions before we proceeded to the area where the supplements were encapsulated.

Unlike many other herbal supplement suppliers, Gaia encapsulates liquid extracts. We got to watch capsules being filled, sealed, inspected, and boxed for delivery over to the packaging warehouse were they are put into Gaia’s patented plant-based bottles.

We spent the rest of our morning touring the farms (they have virtual tours on their website). I got to ask questions about the planting and cultivation techniques, how the crops are treated and harvested, proper climate, etc. It was informative and made me feel as if this is something I could do myself (on a far smaller scale). Additionally, we learned about Gaia’s

And now, for the photos. Imagine standing in field of echinacea and passionflower, surrounded by Gingko and Hawthorn trees, breathing in clean mountain air, with the aroma of relaxing herbs on the breeze. That is what a trip to Gaia is like.

Fields of lemon balm, Melissa officinalis

Passiflora incarnata

Side note: One of the things we learned from Dr. McCoy was that passionflower (the more pink flower in the slideshow) is one of the only herbs found to have been nearly uniformly used by Native American tribes.

Purple Coneflower aka Echinacea Purpurea

As a final, unexpected treat, Gaia treated us to lunch in their dining facility. The food was amazing and fresh but it was the sit down experience that was so memorable. we sat down to outdoor dining with views overlooking the grounds. Truly stunning imagery. We all thought the trip couldn’t possibly get any better and then Ric Scalzo, Founder and CEO of Gaia Herbs, pulled up a chair and ate with us. He generously let us ask him questions related to operations, starting a business, building relationships with suppliers around the world, and so much more. I cannot imagine any other reality where I would have the opportunity to ask a successful CEO to share all his secrets for building a business.

Herbal Ingenuity Located in Wilkesboro, North Carolina, Herbal Ingenuity is a raw material supplier that takes raw botanical ingredients, directly from suppliers, and cleans, cuts, and treats the materials for the end customer. We were able to watch the quality assurance process as workers sifted through the materials, see the machines used for cutting, and ask questions of the staff throughout our time there.

North Carolina Arboretum & Germplasm Repository This part of the trip was the most surprising. When we were initially told we were visiting an arboretum, I was naturally quite happy. What herbalist doesn’t love arboretums? I was entirely unfamiliar with the Germplasm Repository and its unique, important work. Brace yourself, I’m going to gush – and try to respect the confidential nature of some of the work.

Straight from their site, “The North Carolina Arboretum Germplasm Repository (TNCAGR) leads a multifaceted effort to conserve, study and utilize native plants and plant organisms (endophytes).” None of the students were prepared for what we were about to see.

First, we got to meet Dr. Joe-Ann McCoy, the Director of the Germplasm Repository. I took four pages of notes while she spoke. I finally switched to voice clips while she discussed the plight of the Ginseng crop. She generously explained her work in creating voucher specimens, collecting and conserving seeds, encouraging the development of seed banks, researching, grant proposal writing, and conducting research and testing on endophytes – the organisms that live inside of plants. I took an entire page of notes on this alone and am absolutely captivated by topic. By this point in our trip I was already asking Dr. McCoy if took interns.

We got to look around the lab (slideshow below), see the equipment, and gain a greater appreciation for the scientific work being conducted.

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The final part of our time with Dr. McCoy was seeing some of the plants being propagated and a lesson in ethnobotany. She was working with local tribes to propagate plants that had significant traditional and medicinal value to the tribes. Through the trust she had built with them, she had been allowed into a world few were part of. She had developed relationships with numerous native tribes, and was working hard to help them all develop their own voucher specimen collections and herbariums. This unexpected session of the trip was the most inspirational part of the entire trip.

The plant used by the Cherokee, written in Cherokee (top left).

Voucher specimens.

North Carolina State University Mountain Horticultural Crops Research and Extension Center

I took very few notes on this part of the trip because we were hands on with the plants and exploring gardens and woods. We started this visit with a trip to North Carolina State University’s Mountain Horticulture Crops Research and Extension Center. We saw the offices where the staff worked and learned about the Alternative Crops and Organics Program, much of which is dedicated to medicinals. From there we hopped in some trucks and went off to see some of the crops.

The Asian herb garden was fascinating to see. We learned about the current issues in the industry with contaminated Indian and Chinese herbs. The soil in China is full of heavy metals and fecal matter has been found on many plants in India. For this reason many companies are turning to the United States to supply traditional Asian plants. The very unique ecology of western North Carolina is very similar to Asia so the plants can grow successfully without losing the medicinal benefits.

The second half of this trip introduced us to forest farming. We saw some very special and endangered medicinal plants growing in the woods and learned about the ethnobotanical connections between these plants and the local communities. We discussed sustainable harvesting practices, over-harvesting, and ethical issues. Below are some photos of Black Cohosh, Ginseng, Bloodroot, and Goldenseal. Additionally, we learned about the Appalachian Beginning Forest Farmer Coalition.

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Final lesson:

We ended our trip with a lesson from our instructor on sustainable wildcrafting and botanical identification. We visited a state park where we were able to hike and discuss the plants that we encountered.

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ISci 701 Introduction to Statistics, Research Design, and Information Literacy

According to the course description,

This course supports critical analysis of a wide range of integrative health studies. It provides future integrative medicine professionals with the foundational knowledge and skills to identify and evaluate research design and basic statistics. Students develop skills in searching databases as well as critical appraisal of clinical and epidemiological research. Students will find and evaluate published information on health topics then summarize and share their findings.

This course was much more challenging than anticipated. Successful completion of each assignment required demonstration of all of the skills covered in the class to date. An example of the this is the last individual assignment we did prior to the group project presentation. The assignment was to evaluate two scientific studies and determine if they would be clinically relevant to a practitioner’s client population.

ISci 701 Whitney_Palacios_Assignment2