Understanding Jyl Burgener Midwest Area Biosafety Network (MABioN)

MABioN February 18, 2016

Jyl Burgener, M.S. MBA, RBP, CBSP Topics

 1. Definition and Characteristics of Toxins

 2. Regulations Regarding Toxins

 3. Inactivation of Toxins

 4. Human Health Considerations of Toxins

2 Jyl Burgener, M.S. MBA, RBP, CBSP Definition of Toxins

 Simple definition of : a poisonous substance produced by a living thing

 Full definition of toxin: • a poisonous substance • a specific product of the metabolic activities of a living organism • usually very unstable • notably toxic when introduced into the tissues, and • typically capable of inducing antibody formation

3 Jyl Burgener, M.S. MBA, RBP, CBSP Political Definition

 North Atlantic Treaty Organization (NATO) vs Warsaw Pact  Biological or chemical agent?

 NATO opted for biological agent, and the Warsaw Pact, like most other countries in the world, for chemical agent.

 According to an International Committee of the Red Cross review of the Biological Weapons Convention:  "Toxins are poisonous products of organisms; unlike biological agents, they are inanimate and not capable of reproducing themselves",  "Since the signing of the Convention, there have been no disputes among the parties regarding the definition of biological agents or toxins"

4 Jyl Burgener, M.S. MBA, RBP, CBSP Biotoxin

 The term "biotoxin" is sometimes used to explicitly confirm the biological origin.

 Biotoxins can be further classified into:  fungal biotoxins, or short ,  microbial biotoxins,  plant biotoxins,  short phytotoxins and  animal biotoxins.

5 Jyl Burgener, M.S. MBA, RBP, CBSP Characteristics of Toxins

 Extremely potent

 Often mixtures

 Directly damage host tissues

 Disable the immune and nervous systems

6 Jyl Burgener, M.S. MBA, RBP, CBSP Toxins are Extremely Potent

Toxin LD50 Satratoxin 24 ug/Kg 0.02 ug/Kg Botulinum .0003 ug/Kg Cyanide 1000 ug/Kg 2000 ug/Kg

7 Jyl Burgener, M.S. MBA, RBP, CBSP Toxins are Often Mixtures

 Some organisms make a toxin

 Some organisms make multiple toxins  e.g. Penicillium may produce more than 100 different mycotoxins ;  May vary according to specific isolate and prevailing growth conditions.  Stachybotry chartarum also produces numerous compounds: mycotoxins implicated in health effects including macrocyclic trichothecense verrucarins B and J, riridin E. satratoxins F, G, H and isisatratoxins F, G H

8 Jyl Burgener, M.S. MBA, RBP, CBSP US Regulations Regarding Toxins

 Select Agents and Toxin Regulations

 7 C.F.R. Part 3319

 9 C.F.R. Part 12142

 42 C.F.R. Part 73

9 Jyl Burgener, M.S. MBA, RBP, CBSP International Regulations: Biological and Toxin Weapons Convention (BWC)

 Supplements the 1925 Geneva Protocol  Prohibits use of but not possession or development of chemical and biological weapon

 173 countries are party to it (Dec. 2014)  Prohibit development, production and stockpiling of biological and toxin weapons

 No formal verification method to monitor compliance limits effectiveness of the convention

 U.S. Congress passed the Bioweapon Anti-Terrorism Act in 1989 to implement the BWC

10 Jyl Burgener, M.S. MBA, RBP, CBSP Inactivation of Toxins – Guidance

 Appendix I—Guidelines for Work with Toxins of Biological Origin

 Prudent Practices in the Laboratory –  Ch.6 Working with Chemicals –  Ch. 6.D. Working with Substances of High Toxicity

 OSHA Lab Standard – Particularly Hazardous Substance

11 Jyl Burgener, M.S. MBA, RBP, CBSP Inactivation of Toxins: Steam

 Steam Heat (Autoclaving)  Generally, many of the higher molecular weight, proteinacious bacterial biotoxins can be inactivated by autoclaving for 1 hour at 121°C.  Autoclaving should not be used for destruction of low molecular weight biotoxins such as mycotoxins, marine and reptile .

12 Jyl Burgener, M.S. MBA, RBP, CBSP Steam Autoclaving (121 oc/15 psi/1 hr)

 Abrin 

 Botulinum  Pseudomonas A

 Cl. difficile – binary toxin c,  Ricin A, Toxin A, Toxin B  Shigatoxin & Shiga-like ribosomes inactivating  Cl. Perfringes (epsilon) proteins

 Cl. Tetani (tetnus toxin)  Staphylococcal

 Diptheria toxin

13 Jyl Burgener, M.S. MBA, RBP, CBSP Inactivation of Toxins: Chemical

 Chemical Inactivation –  Sodium Hypochlorite (NaOCL) – use a 2.5% solution, equivalent to a 50% Bleach solution. Minimum of 30 min. exposure.  Sodium Hydroxide (NaOH) – 1 N; Minimum of 1 hour exposure (Prions) usually hold overnight

14 Jyl Burgener, M.S. MBA, RBP, CBSP Chemical Inactivation – 2.5% NaOCL (w/v)

 Anthrax Lethal/Edema  Ricin Toxin (PA & LF)/(PA & EF)   Botulinum neurotoxins  Staphylococcal  Brevetoxin enterotoxin

 Diptheria toxin  (TTX)

 Microcystines  T-2

 Palytoxin

15 Jyl Burgener, M.S. MBA, RBP, CBSP Chemical Inactivation – NaOH 1 N 1 hr

 Botulinum

 Staphylococcal Enterotoxin

16 Jyl Burgener, M.S. MBA, RBP, CBSP Inactivation of

 Depends on the presence or absence of disulfide bonds.

 Conotoxins with disulfide bonds will be incubated with an excess of dithiothreitol (DTT) (usually 10-20mM buffered solution, pH8.7) for at least 1 hour at room temperature or 30 minutes at 50oC.  Then, an equal volume of 50-100mM solution of iodoacetamide (buffered solution, pH8.7) will be added and incubated for 1 hour at room temperature.

17 Jyl Burgener, M.S. MBA, RBP, CBSP Human Health Considerations of Mycotoxins

 80% of Clinical Toxin exposure is mold/mycotoxin exposure.  Affects more humans than any other toxin  More detailed information is known

 25% of the population has an inability to process/clear mold toxins which leads to a series of biochemical alterations in their bodies

 322 mil x .25% = 80.5 mil people

 There are many mold symptoms, because the illness affects multiple systems in the body

 Not all molds are toxigenic, but Stachybotrys, Penicillium, Aspergillus, and Chaetomium are the most common toxigenic ones.

18 Jyl Burgener, M.S. MBA, RBP, CBSP Symptoms of Mycotoxin Exposure

 Fatigue ♦Weakness ♦Aches ♦Muscle Cramps ♦Unusual Pain ♦Ice Pick Pain ♦Headache ♦Light Sensitivity ♦Red Eyes ♦Blurred Vision ♦Tearing ♦Sinus Problems ♦Cough ♦Shortness of Breath ♦Abdominal Pain ♦Diarrhea ♦Joint Pain ♦Morning Stiffness ♦Memory Issues ♦Focus/Concentration Issues ♦Word Recollection Issues ♦Decreased Learning of New Knowledge ♦Confusion ♦Disorientation ♦Skin Sensitivity ♦Mood Swings ♦Appetite Swings ♦Sweats (especially night sweats) ♦Temperature Regulation or Dysregulation Problems ♦Excessive Thirst ♦Increased Urination ♦Static Shocks ♦Numbness ♦Tingling ♦Vertigo ♦Metallic Taste ♦Tremors

 The typical patient will exhibit an average of 23 symptoms

19 Jyl Burgener, M.S. MBA, RBP, CBSP Human Health Effects - Detection

 Mycotoxins: the direct cause of diseases (eg asthma), liver and kidney cancer, permanent genetic damage, brain damage, nervous system damage and even death

 Mycotoxins can be detected in urine

 Blood tests can be used (measure MMP9 and TGF-beta and C4a in particular)

 Genetic testing

 Visual Contrast Sensitivity Test

 Neuro Quant

20 Jyl Burgener, M.S. MBA, RBP, CBSP HLA Genetics

 Currently 12 genes associated with the clearance of toxins from the body  Located on Chromosome 6

 Do not allow an individual to eliminate circulating biotoxins

 Bioaccumulation occurs

 Patient develops chronic illness

 Genotype –HLA-DR found in ~25% of population

 ~ 90% of patients developing chronic Lyme disease and chronic ciguatera poisoning have HLA-DR

21 Jyl Burgener, M.S. MBA, RBP, CBSP Visual Contrast Sensitivity

 Ken Hudnell (neurotoxicologist) was the first to use VCS to measure the effects of biotoxin exposure and heavy metal toxicity and as a marker for neurotoxic exposures.  VCS testing has also revealed neurologic deficits associated with exposure to toxic Pfiesteria sp. and other biotoxins ( Ciguatera ;Chronic Ciguatera Seafood Poisoning), cyanobacteria such as Cylindrospermopsis and Microcystis, various species of fungi such as Stachybotrys, Aspergillus, Penicillium, and Fusarium, and Brown Recluse spider as well as Borrelia burgdorferi (Lyme disease organism)

22 Jyl Burgener, M.S. MBA, RBP, CBSP Visual Contrast Sensitivity (VCS) Test

 92% of people with biotoxin illness have a positive VCS test

 VCS has been a sensitive indicator of neurotoxicity induced by organic solvents.  Persistent or permanent VCS deficits in the presence of normal visual acuity have been observed in styrene and mixed-solvent exposed workers in the absence of detectable optical, retinal or optic nerve head pathology.

23 Jyl Burgener, M.S. MBA, RBP, CBSP NeuroQuant Brain MRI

 A volumetric MRI which measures regional brain structures known to deteriorate in patients with Alzheimer’s disease and other related neurodegenerative disorders.

 NeuroQuant is the only volumetric MRI application to use an FDA approved database of normal same age peers for comparison.

24 Jyl Burgener, M.S. MBA, RBP, CBSP NeuroQuant Brain MRI

 People with elevated TGF-beta and MMP9. show in NeuroQuant studies a swelling of the frontal lobes, the hippocampus and the cerebellum and a shrinkage of the caudate.

 The frontal lobe is where the executive function is found.

 The hippocampus is associated with mood.

 The cerebellum is associated with movement disorders.

 The caudate is very dopamine rich and is highly associated with moods and feeling good.

25 Jyl Burgener, M.S. MBA, RBP, CBSP Common Mycotoxins

 Aspergillus: B1 –  Liver cancer

 Fusarium: Zeatralenone;  Estrogen disturbance

 Penicillium  Tremogenic mycotoxins

 Stachybotrys; tricothecenes  Neurotoxins

26 Jyl Burgener, M.S. MBA, RBP, CBSP Hormonal Indicators

 Toxins impact the adrenals, thyroid, and other hormones.  ADH (anti-diuretic hormone) deficiency is almost universal in biotoxin illness

 When ADH is low, you lose water.  Salt then gets dumped onto the skin via the sweat glands. Dr. Shoemaker found that when people experienced static shocks, they had high chlorides

 When MSH is low, ADH is generally low.  Reduced MSH is a co-factor which can lead to MARCoNS can also produce biofilms and biotoxins

27 Jyl Burgener, M.S. MBA, RBP, CBSP Biotoxin Illnesses

 Biotoxin Illness is also called Chronic Inflammatory Response Syndrome (CIRS),  The body loses the ability to create antibodies to mold toxins.

 The acquired immune system of 24% of the population has susceptible genes breaks  Not everyone with susceptible genes has trouble making antibodies to mold inflammagens.

 According to the work of Dr. Shoemaker:  Inflammatory illnesses like Mono, Lyme, Coxsackie, lung inflammation, or other inflammatory diseases can set the stage for a breakdown in proper antibody creation.

28 Jyl Burgener, M.S. MBA, RBP, CBSP Recirculation of Toxins

 The toxins are reabsorbed by a recycling process in the body called enterohepatic circulation that serves to reuse bile and bilirubin.

 As a result, the toxins are never eliminated and end up back in the bloodstream.

 In biotoxin ill people, the toxins perpetually circulate throughout the body and keep the innate immune system in overdrive resulting in massive inflammation.

 Note: It’s actually the increase in innate immune system constituents that produce symptoms when you get sick – fatigue, aches, brain fog, etc.

29 Jyl Burgener, M.S. MBA, RBP, CBSP Air Sampling – Accepted Practices

 3 IAQ samples, area of complaint, area on non-complaint and outside

 Misinterpretation

 Results can be skewed

 Taking the results down to only the genus level –  Insufficient data for litigation

30 Jyl Burgener, M.S. MBA, RBP, CBSP Environmental Relative Moldiness Index (ERMI) Testing

 An objective, standardized DNA based method of testing that will identify and quantify molds.

 Uses the analysis of settled dust in homes and buildings to determine the DNA concentrations of the DNA of different molds.

 ERMI scale ranges from -10 up to 20.

 ERMI rating of < 2 good

31 Jyl Burgener, M.S. MBA, RBP, CBSP Psychiatric Disorders

 Cytokines are associated with treatment-resistant depression

 Cytokines have been shown to increase the permeability of the blood-brain barrier membrane

 Vaccines given to depressed patients do not mount a good immune response

32 Jyl Burgener, M.S. MBA, RBP, CBSP Psychiatric Disorders, cont’d

 Some studies by psychologists show clear cognitive impairment associated with mold

 Mycotoxins excreted by mold are clearly neurotoxic

kill off the olfactory neurons.  May be a reason for multiple chemical sensitivity in so many people

 T-2 (Fusarium) kills normal brain cells indiscriminately

(Aspergillus) depletes the striatal dopamine  Associated with mood disorders and also movement disorders such as Parkinson’s

33 Jyl Burgener, M.S. MBA, RBP, CBSP References

 Ritchie C. Shoemaker, Dennis House, James C. Ryan, Structural brain abnormalities in patients with inflammatory illness acquired following exposure to water-damaged buildings: A volumetric MRI study using NeuroQuant􏰍􏰍, Neuro- toxicology and Teratology (2014), doi: 10.1016/j.ntt.2014.06.004  SurvivingMold.com  Benros ME, Waltoft BL, Nordentoft M, Ostergaard SD, Eaton WW, Krogh J, Mortensen PB. Autoimmune diseases and severe infections as risk factors for mood disorders: a nationwide study. JAMA Psychiatry, 2013 Aug; 70(8): 812-20.

34 Jyl Burgener, M.S. MBA, RBP, CBSP