Making Honey and Medicine a Local Affair Part 1: The Clinical Trial: Dr. Stephen Rankin (Pinon Family Practice) Part 2: The Honey Analysis: Dr. Don Hyder (San Juan College) Dr Stephen Rankin and Dr. Don Hyder The Clinical Trial New Mexico Honey Wound Treatment Research Study Approvals FDA has approved the project, IND SJRMC IRB has approved the Honey Project SJC IRB has approved the Honey Project in relation to the Distinguished Teaching Chair Award Clinical Trial Planning Committee R. Stephen Rankin, M.D., Principal Investigator Joseph Pope, M.D. Mary Doshi, Associate Professor, SJC, Medical Laboratory Don Hyder, Professor, SJC, Biology Lynn Lane, Technology Trainer, SJC, Computer Science Beth Philips, Research Consultant, SJRMC IRB Administrator History Honey has been used for wound healing for over a thousand years Honey used in wound care comes from Australia and New Zealand Samples from Ireland and Africa tested against CA-MRSA (community acquired MRSA) isolates and other Staphylococcal specimens Results were favorable in demonstrating activity against CA- MRSA Little in vitro or in vivo testing of honey from the United States Preliminary in vitro studies from Northwest New Mexico honey demonstrated bactericidal activity against CA-MRSA Preliminary screening of the local honey for C. botulinum Benefits of honey in wound healing Moist, antibacterial wound healing environment Prevents spreading of infection Renders wound sterile Rapidly eliminates wound odor Cleans and debrides wound Anti-inflammatory No adverse effects Diminishes pain Reduces swelling Works when other traditional treatments have failed caMRSA* Community acquired Methicillin Resistant Staphylococcus aureus Common cause of skin and soft tissue infection May become invasive to brain, spinal cord, bone, and other body tissues High incidence of recurrences personal and family In Vitro Studies Acquired MRSA Manuka Honey Local Project Honey Lot 1 Vancomycin Wildflower Local Project Honey Lot 2 Clover Control Photo courtesy of Mary Doshi,MLS, (ASCP) Director, Program in Medical Technology, San Juan College Our study Further investigate in vitro activity of this New Mexico varietal honey against CA-MRSA Collaboration of Pinon Family Practice, San Juan College, and SJRMC Conduct a controlled clinical research protocol demonstrating its effectiveness against CA-MRSA abscesses Involve students at San Juan College from multiple disciplines Patient population characteristics Inclusion: Ages Abscess less than or equal to 6 cm in diameter on arms, legs, abdomen, back Patient willing to consent and try honey prepared dressing Patient willing to return to office each day for 7 days Exclusion: Under age 16 or 80 or above Have underlying immune system disorder Abscess of face, scalp, breast or genitals, hands or feet Diagnosed as diabetic Pregnancy Allergic to bee pollen, honey, sulfa antibiotics, or lidocaine Diagnosed with peripheral vascular disease Recurrent use of alcohol or drugs Study procedure After consent patient receives incision and drainage of abscess and randomized to : 1)Topical honey 2)Oral antibiotics Culture of wound sent to SJRMC and SJC on Day 1 Daily photos of wound Measurement of wound size Temperature Daily cultures of wound sent to SJC Return daily for 7 days Goals Compare efficacy Compare side effects Compare sterilization of wound Compare patient acceptance of treatments PRELIMINARY RESULTS Wound (Erythema) Antibiotic patients (N = 7) Average initial size in cm squared 27.9 (range 6.6 to 72.0) Average final size in cm squared 9.7 (range 0.0 to 42.0) 65% reduction Honey patients (N = 4) Average initial size in cm squared 16.0 (range 3.2 to 36.0) Average final size in cm squared 2.6 (range 0.0 to 9.0) 83% reduction Skin and soft tissue infections Current treatment involves incision and drainage with or without antibiotics Role of antibiotics is uncertain with attendant side effects and resistant organisms Recurrent infections with MRSA are significant Study by Fritz (Clin. Infect. Dis. 2012:54: ) showed a 72% recurrence rate at 12 months even with decolonizing regimen Clearly something else is needed Can a topical bactericidal honey have a role in both treatment and preventing recurrences The bees Treatment Free (No antibiotics or miticides) Survivor Queens locally raised and from breeders who use no treatments Forage is varied, along San Juan River near Farmington, New Mexico Honey is unheated and unfiltered Characterization of Antibacterial Components, Pollen and Nectar Sources of a Honey Being Used to Treat Community Acquired MRSA Infections Bryden Baker, Holly Vandever, Dr. Eric Miller and Dr. Don Hyder Funding provided by the National Institute of General Medical Sciences (8 P20 GM ) from the National Institutes of Health Hypothesis Locally produced clinical honey currently being investigated in the treatment of Community Acquired Methicillin Resistant Staphylococcus aureus (CA-MRSA) is derived from a single pollen source (Russian Knapweed) and the primary antibacterial compound is methylglyoxal. Methylglyoxal is known from a medically recognized honey, Medi- honey or Manuka Honey, derived from the Manuka plant in New Zealand. Objectives 1) Identify the primary floral source, (pollen and nectar) of the local clinical honey 2) Identify and quantify a known antibacterial component of honey, methylglyoxal 3) Investigate the antibiotic components of nectar from Russian Knapweed Methods (Floral Sources) 1) Survey and collect pollen from floral sources in area surrounding Study Area bee hives 2) Prepare and observe pollen using light and electron microscopy 3) Extract pollen from clinical honey and observe using light microscopy 4) Quantify pollen in clinical honey using a pollen trap on bee hives Methods (Chemical Analysis of Clinical Honey, Manuka Honey and Russian Knapweed Floral Extract) 1) Prepare Clinical honey, Manuka honey and Russian Knapweed floral extract for extraction using Solid Phase Micro-extraction (SPME) techniques. 2) Analysis of extracts using Gas Chromatography-Mass Spectrometry Pollen Study Results Russian Knapweed Coyote Willow Musk Thistle Coyote Willow TamariskFourwing Saltbush Russian Knapweed Pollen Grains Using Light Microscopy Pollen Study Common Name *1 Scientific name Pollen Grain Size (nm) Pollen Trap (color and percent of pollen balls per 20 ml sample) Pollen Identified in Clinical Honey Russian Knapweed Centaurea repens40 Brown/Tan- 37% * Four-wing saltbush Atriplex canescens30 Bright Yellow 14% TamariskTamarix parviflora20Rust- 2%* Tansy MustardDescurinia pinnata40 Princess PlumeStanleya pinnata20-40 Russian Olive Elaeagnus angustifolia 40 Coyote WillowSalix exigua17-20White 32% * Showy MilkweedAsclepias speciosa40 Curly Cup Gumweed Grindelia squarrosa Musk ThistleCarduus nutans40Dark Yellow 15% and Orange-0.5% * *1 : Pollen collected from plants blooming in the vicinity of the bee hives at the same time as the Russian Knapweed. Pollen Study Summary The clinical honey was identified as a multiple floral source honey with Russian Knapweed pollen and Coyote Willow pollens being significant components along with four other pollen sources. Chemical Analysis Results CompoundsRTNoMW (g/mol) Methylglyoxal Acetic anhydride ,2-Propanediol ,3-Dioxolane-2-methanol Methyl-1,6-heptadien-4-ol Methyl 6-oxoheptanoate ,2-Dimethyl-1,3-dioxan-4-yl ethanol Silane Hexenal Hexanedioic acid Diethyl 1,2-cyclopropanedicarboxylate D-allo-dec-2-enonic acid Methyl-1-6-ethyl-3-octyloxy-1-silacyclohexane Manuka Honey CompoundsRTNoMW (g/mol) Methylglyoxal Furfural ,3,5-Cycloheptatriene H-Pyran-4-one Furancarboxaldehyde Dodecanol Morphinan-3,14-diol Glucopyranoside Clinical Honey Russian Knapweed Floral Extract CompoundsRTNoMW (g/mol) R-(-)-2-Amino-1-propanol1.341 Topotecan Benzenemethanol, a-(1- aminoethyl)-,[S-(R*,R*)] ,5-Furandicarboxaldehyde5.897 N-Benzoyl-dl-alanine ,1-(4-Methyl-1,3- phenylene)bis[3-(5-benzyl- 1,3,4-thiadiazol-2-yl)urea (+)-Ascorbic acid 2,6- dihexadecanoate Pregan-20-one, 2-hydroxy- 5,6-epoxy-15-methyl ,12,15Octadecatrienoicacid Retinal 9-cis Chemical Analysis Results Summary 1) Methylglyoxal was identified in the clinical honey and in the Manuka, Medi-honey. 2) A secondary compound, furfural, a phenolic compound having known antibacterial and antifungal properties was identified in the local clinical honey. 3) A known anticancer compound, topotecan (A topoisomerase inhibitor), was identified in the Russian Knapweed floral extract. Antibacterial compounds were not found in the Russian Knapweed floral extract. Further Research Continued Investigation of Floral source(s) of this local New Mexico honey Chemical analysis and in-vitro studies of other local and regional honey samples against MRSA Further clinical research involving bactericidal honey and MRSA recurrences Quantify chemical components of the honey such as methylglyoxal, furfural and bee definsin-1 levels Contact Information Dr. Stephen Rankin: Dr. Don Hyder: