Chronic Candida – The Role of Biofilm in Candida Pathogenicity and

Treatment Options

Copyright© 2015, Kurt N. Woeller, D.O., Tracy Tranchitella, N.D., and Educational Resource Association. This material may not be reprinted,

distributed or used without permission.

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Lecture Overview

•What are biofilms?

•How and why are they created?

• The role of biofilms in Candida pathogenicity.

• The role of biofilms in bacterial infections.

• Expanded treatment options

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Support Documents For Module #4

• Biocidin Biofilm Study (pdf)

• Virulence Article – Candida Pathogenicity (pdf)

• Biofilm Program – Patient Handout

• Biofilm Program – Handout Example

• Lecture slides (pdf)

• Lecture slides - note taking (pdf)

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What Are Biofilms?

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Over 500+ biofilm species colonize the human mouth, causing tooth decay and gum disease. This image, retrieved infected dental implant, was taken by a scanning electron microscope.

Source: Rachel Sammons, PhD, University of Birmingham, Dept. of Biomaterial.

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Various Disorders Associated With Biofilms

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Source: Center for Biofilm Engineering at MSU - Bozeman

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Associated Disorders

• Atherosclerosis

• Endocarditis

• Chronic sinusitis

• Inner ear infections

• Osteomyelitis

• Prosthetic joint and heart valve involvement

• Urinary tract infection

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Viridans streptococci

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“The clinical impact of bacterial biofilms.” Hoiby N, et.al. Int. J Oral Sci. 2011 Apr;3(2):55-65.

“Pseudomonas aeruginosa biofilms in cystic fibrosis.” Ciofu O, et. al. Future Microbiol. 2010 Nov;5(11):1663-74.

“The role of bacterial biofilms in chronic infections.” Bjarnsholt T. APMIS Suppl. 2013 May;(136):1-51.

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Pseudomonas aeruginosa and Cystic Fibrosis

P. aeruginosa in trachea P. Aeruginosa – forming biofilm

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Lung of Cystic Fibrosis patient with P. aeruginosa biofilm

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Gordon Ramage, et.al. “Candida Biofilms: an Update” Eukaryot Cell. 2005 Apr; 4(4): 633–638.

• “Candida albicans remains the fungal species most commonly associated with biofilm formation.”

• “…formation of Candida biofilms carries important clinical repercussions because of their increased resistance to antifungal therapy and the ability of cells within biofilms to withstand host immune defenses.”

• “Biofilm formation on medical devices can negatively impact the host by causing the failure of the device and by serving as a reservoir or source for future continuing infection.”

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Candida Biofilm on Catheter and Plastic

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Candida Hypha Transformation and Biofilm Development

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François L. Mayer, et. Al. Candida albicanspathogenicity mechanisms. Virulence. Feb

15, 2013; 4(2): 119–128.

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Candida Transformation

• To infect host tissue, the unicellular form of Candida switches to a more invasive form based on environmental stressors.

• Examples: pH, food supply, temperature, CO2 at the tissue level.

• Candida can control its environment too to become invasive.

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Candida Transformation and Biofilm Formation

• Heat shock proteins (HSP) – these specialized proteins are designed to prevent against protein unfolding and aggregation, caused by environmental stress, which normally induces cell death. HSPs are involved in Biofilm formation, morphogenesis, virulence, and drug resistance too.

• Metal accumulation – Candida acquires iron, zinc, manganese and copper for metabolic survival, with iron and zinc as primary minerals.

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Adhesins and Invasins

• Candida has specialized proteins which allow it to adhere to other microorganisms, abiotic surfaces, host cells and tissues (biotic surfaces). These adhesion proteins are also involved in Biofilmformation.

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Candida Biofilm Formation

Candida biofilms form in a sequential process:

1. Adherence of yeast cells

2. Proliferation

3. Hyphal cell formation in upper biofilm layer

4. Extracellular matrix material accumulation and dispersion of yeast cells from biofilm.

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Candida Biofilm Development and Dispersal

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Contact Sensing

• Contact with certain surfaces will induce yeast cells to switch to hyphal transformation and biofilm formation.

• Thigmotropism – different topography, i.e. ridges will induce hyphal invasiveness damaging adjacent cells and tissues.

• Secreted hydrolases – protease, phospholipases, and lipases.

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APMIS Suppl.2013 May;(136):1-51“The role of bacterial biofilms in chronic infections.”

Bjarnsholt, T. et.al.

• “Bacteria have two life forms during growth and proliferation. In one form, the bacteria exist as single, independent cells (planktonic) whereas in the other form, bacteria are organized into sessile aggregates. The latter form is commonly referred to as the biofilm growth phenotype.”

• “Acute infections are assumed to involve planktonic bacteria, which are generally treatable with antibiotics, although successful treatment depends on accurate and fast diagnosis.”

• “In cases where the bacteria succeed in forming a biofilm within the human host, the infection often turns out to be untreatable and will develop into a chronic state.”

• “The important hallmarks of chronic biofilm-based infections are extreme resistance to antibiotics and many other conventional antimicrobial agents, and an extreme capacity for evading the host defenses.”

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• Although enteric biofilms are likely in a steady state of shedding and regrowth, and although the percentage of epithelium covered with biofilms is unknown…

• “There is no doubt that biofilms are indeed present in the normal bowel.”

• It is not overly surprising to observe biofilms in the normal colons of humans since similar observations have been made in the normal colons of laboratory animals as diverse as mice.

Gut. 2007 Oct; 56(10): 1481–1482. “Biofilms in the normal human large bowel: fact

rather than fiction” R Randal Bollinger, et.al.

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Biofilm Resistance

Biofilms have different levels of resistance:

1. Complex architecture of biofilm

2. The biofilm matrix

3. Efflux pumps

4. Metabolic plasticity

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Source: Center for Biofilm Engineering, Montana State University-Bozeman

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Quorum Sensing

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Quorum Sensing

• The ability of a single organism, i.e. yeast or bacteria to sense how many other organisms are in its vicinity and allowing it to join up with the biofilm colony.

• Can occur amongst a specific species or varied species. For example, different bacteria or bacteria and yeast.

• P. aeruginosa – and the creation of biofilms in cystic fibrosis.

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Staphylococcus Biofilm

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Source: Center for Biofilm Engineering, Montana State University-Bozeman

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Clostridia bacteria – spore forming

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Blend of Different Pathogens

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Treatment Options for Candida Biofilm

(and bacteria too)

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Observed Clinical Indicators:

Lack of stool culture findings for yeast and/or bacteria:• However, by clinical history, individual seems to improve

when placed on antibiotics or antifungal medication or botanicals.

No Improvement With Antifungal or Antibacterial Treatment – Think Biofilm Problem

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Individuals with Obsessive-Compulsive Disorder (OCD), or other MH or physical problems:• Who tend to get worse seasonally or worsen with strep

exposure (may or may not test positive), or who have recurrent infections despite benefit of antibiotic treatment.

Persistent yeast and/or bacteria infections despite treatment:• As evidenced on Organic Acids Test such as arabinose

for yeast or HPHPA and/or 4-cresol for clostridia bacteria.

No Improvement With Antifungal or Antibacterial Treatment – Think Biofilm Problem

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General Recommendations for Implementing Biofilm Program

• Avoid implementing biofilm program for a ‘new’ parent just getting starting with anti-pathogen treatment.

• Determine how sensitive patient is to initial antimicrobial remedies.

• Expect “die-off” with initial implementation of biofilm supplements. Go slow at first:• Start with before breakfast dose of enzyme for 5 to 7 days before

adding a before lunch or dinner dose.

• Also, consider starting ½ dose for sensitive individuals.

• Generally, biofilm program should be reserved for parents with experience in giving antimicrobials to their kids.

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• Step #1 – Lysis (breakdown) and Detachment of Polysaccharides.• plus additional interventions, i.e. Stevia

• Step #2 – Attacking the Microbes

• Step #3 – Cleaning Up

• Step #4 – Additional considerations for long-term gut support and potential biofilm formation inhibition.

Biofilm Program – Basic Steps

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Biofilm Enzymes - examples

Interfase Interfase Plus

2 to 8 capsules daily between meals1 to 4 capsules daily between meals

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Step #1 – Lysis & Detachment

• Enzymes (good option is Interfase Plus)• Another option = Interfase (w/o EDTA) for sensitive

individuals.

• NaEDTA (helps to bind minerals in biofilm)

• Taken on empty stomach 30 minutes before or 90 minutes after meals (breakfast, lunch, dinner).

• Polysaccharides = complex sugars with mineral linkages – iron, calcium, magnesium.

• Take minimally 30 minutes before Step #2

Biofilm Enzymes - example

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Additional Consideration to Enhance Biofilm Breakdown

• Stevia – is a sugar substitute and sweetener taken from the plant species Stevia rebaudiana.

• Reported to help in the degradation of biofilms by opening up fluid channels which allows for antimicrobial remedies to penetrate deeper into the biofilm matrix.

• Good options for kids as it can be added to dilute juice.

• Full dose = 7 drops (reported for Lyme?). For kids, consider using 2 to 4 drops on empty stomach.

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Step #2 – Attacking the Microbes

• Antibiotics – Flagyl, Vancomycin, Alinia, Penicillin, Zithromax, etc.

• Antifungals – Nystatin, Diflucan, Sporanox, Lamisil

• Natural Remedies:• Biocidin, Oil of Oregano, Olive Leaf Extract, Uva Ursi,

Caprylic Acid, Allimax, Candida Yeast Packet, other individual or combination botanicals.

• These are taken approximately 30 minutes after Step #1 biofilm enzyme product is administered.

Biofilm Program

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Additional Biofilm Remedies

• Lactoferrin – binds to iron which is an important mineral for candida and bacteria.

• Essential plant oils, i.e. Biocidin – oils reported to turn off ‘quorum sensing’ and pump system, and disrupt biofilm formation.

• Curcumin (turmeric) + Quercetin Complex –Curcumin aides in decreasing inflammation, liver and colon support; whereas the Quercetin helps with decreasing excess histamine production.

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1 to 2 capsules QD ½ to 1 capsule BID to TID

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USP Effectiveness Testing of Biocidin®

Demonstrates inhibitory activity of against both gram positive and gram negative bacteria, yeast and fungus.

Bilberry extract, Grape Seed extract, Milk

Thistle, Echinacea, Goldenseal, Shiitake,

White Willow, Garlic, Black Walnut (hull

and leaf), Raspberry, Fumitory, Gentian,

Noni, Tea Tree oil, Galbanum oil,

Lavender oil, Oregano oil

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Biocidin Advanced Formula

Binghamton University researcher Claudia Marques, Ph. D. – see articleIn Module 5 titled “Biocidin Biofilm Study 2013”

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Step #3 – Cleaning Up

• Give remedies 60 to 90 minutes after Step #2

• Activated Charcoal – ½ to 2 capsules

• Fiber, i.e. TruFiber, Apple Pectin, Ground Flax Seeds – ½ to 1 tablespoon.

The main reason for the Activated Charcoal (AC) is to bind “die-off” toxins generated by Step #1 and #2. If individual is not experiencing “die-off” than AC is not absolutely necessary. However, a fiber product is worthwhile to keep the stool

consistency bulky and prevent against constipation.

Biofilm Program

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Binding Agents (Charcoal) and Prebiotic “Fertilizer”

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Biofilm Program

Step #4 – Preventing Biofilm Formation

½ to 1 capsule between meals 1 softgel 1 to 2 times daily w/food –cis-2-decenoic acid helps to dispersepolysaccharides involved in biofilm

formation.

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Mild Candida Protocol

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Moderate Candida Protocol

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Severe Candida Protocol

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Typical “Die-Off” symptoms

• Headache

• Nausea

• Fatigue

• Irritability

• Moodiness

• Digestive bloating and gas

• Fever

• Diarrhea or mixed bowel, i.e. constipation and loose stools

• Rashes

• Body itching

• Hyperactivity and increased self-stimulatory behavior

• Sleep disturbance

See “Die-Off Reaction Management” document in Module 3 for supplement and dosing suggestions.

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Topic

• “The Adverse Chemical Influences of Clostridia Bacterial Toxins and Chronic Health Problems –Beyond Common GI Issues”• Clostridia bacteria toxins and how they negatively affect

the nervous system

• HPHPA & 4-cresol as the main culprits for dopamine enzyme inhibition.

• The problems with excess dopamine

• A deeper understanding of Probiotics

• Treatment options – medications, natural remedies, and protocols.

Module #5

Thank YouKurt N. Woeller, D.O.

www.GIMasteryCourse.com

GIMastery@gmail.com