Blue and Grey

Anthrax

MIlad khourySpring 2011

ARTICLE

New medical weapons to protect against Anthrax attacks

Bouzianas et al. Current and Future Medical Approaches To Combat the Anthrax Threat. Journal of Medicinal Chemistry, 2010; 53 (11): 4305 DOI: 10.1021/jm901024b

Outline

I IntroductionII History of the diseaseIII Its use as a biological weaponsIV Bacillus anthracis V the 3 routs of inectionVI PathogenesisVII The fight against anthraxVIII Conclusion

Anthrax: basics

From the Greek word anthracis for coal.

caused by the bacteria B.anthracis

Zoonotic disease in herbivores (e.g., sheep, goats, cattle)

Human infection typically acquired through contact with anthrax-infected animals or animal products .

Three forms

-Cutaneous -Inhalational -Gastrointestinal

history

1500 B.CMoses threatened the Egyptian pharaoh that the hand of the lord will fall on the livestock ,horses and birds and kill everything.

5th century B.Cthe Greek Doctor Hippocrates named the disease that caused black skin lesions as anthracis.

history

Early 1800s - The first human cases of cutaneous anthrax in the US and UK were reported in men who contracted the disease after having been in contact with infected livestock.

The disease was also called Wool Sorters disease and Rag Pickers disease .

1876 - German bacteriologist Robert Koch confirmed bacterial origin of anthrax.

1881: Louis Pasteur developed the first vaccine

Anthrax a biological weapon

1915 - German agents injected horses, mules, and cattle with anthrax during WWI. This was the first recorded use of anthrax as a biological weapon.

1979 - In Soviet Union, aerosolized anthrax spores were released accidentally at a military facility, affecting 94 and killing 64 people.

1995 - Iraq produced 8,500 liters of concentrated anthrax as part of the biological weapon program under Saddam Husseins administration.

Anthrax cases 2001

22 cases-11 cutaneous-11 inhalational

5 deaths ( all inhalational )-case in Florida-2 postal workers in Maryland

-hospital worker in NY city

Farm woman in Connecticut

Bacillus anthracis

Gram positive rod

has a diameter of 1-1.5 m and a length of 3-10 m.

Facilitative anaerobe

Belogns to the B.cereus family - Nonmotile -glutamyl-polypeptide capsule -soil dwelling

Bacillus anthracis

1200 strains distributed worldwide

Forms spores -infectious form -1micrometer in size

Vegetative state -non infectious - fragile

The spore

Sporulation conditions: -starvation- presence of oxygen-changes in environmental conditions

- resistant to cold, heat,dryness and chemicals

-lethal dose of 2500 to 5500 spores

- spore viability increases in soil with organic content, alkaline PH, high calcium concentration ,high moisture

-

Spore anatomy

Cr: core

Cx: cortex

Ct: coat

Is: interspace

Ex:exosporium

Thin-section electron micrograph

Spore anatomy

The core: -protects the spore from Uv radiation and stress-forms the center part of the spore- chromosomes form complexes with small acid soluble proteins abbreviated as SASP-high levels of dipicolinic acid and ions

The Cortex- important in providing resistance to the spore and keeping the core dry.

Spore anatomy

The coat- multilayer protein shell that prevents the entry of large degradative molecules and microbe predators into the spores core- flexible ridges that unfold, allowing the core's volume to increase when water enters the spore during germination.

The exosporium -forms the outermost structure of the spore that interacts with the environment

-This protein shell is composed of surface proteins

The anthrax cycle

Cutaneous anthrax

Infection of the skin due to direct contact with B.anthracis

contact with animal hides or hair, bone products, and wool.

Stages -primary lesion appears as papule similar to an insect bite-papule enlarges,small vesicles form on its surface

-the vesicles fill up with neutrophils and bacilli. -the vesicles undergoe necrosis and form painless alcer surrounded by an eschartr

Cutaneus anthrax

Most common naturally occurring form

Case fatality after infection -untreated 20 %-with antimicrobial therapy 2%

Complications infection can spread into blood stream and cause shock and death.

Treatmentdoxycyclene antibiotic for 60 days

Cutaneus anthrax

Gastrointestinal anthrax

Caused by the ingestion of meat contaminating Bacillus anthracis

Uncommon in the US

Case fatality rate:50-75%

2 different forms of GI anthrax 1) oral- pharyngeal 2) abdominal

Gastrointestinal anthrax

Oral -pharyngeal form results from the deposition and germination of spores in the upper GI tractcauses infection of lymph glands and lymph channels,edema,sepsis

Abdominal form -develops from the disposition and germination of spores in the lower GI tract.-causes gastroenteritis and symptoms such as vomiting and abdominal pain

Gastrointestinal anthrax

Abdominal anthrax

oral-Pharyngeal anthrax

Inhalation anthrax

The most lethal type of anthrax

Caused by the inhalation of anthrax spores.

Initial phase: - nonspecific, flue like symptoms, fever and malaise

Second phase:-respiratory distress-dyspnea, cyonosis ,mediastinal widening and death

Case fatality :75-90% if untreated

Inhalation anthrax

Inhalation anthrax

Diagnosis:- Blood cultures- Chest x-ray or CT scan of the chest- Sputum cultures

Treatment: Inhalation anthrax is usually treated with intravenous (IV) ciprofloxacin plus another antibiotic sixty days

Complications: Hemorrhagic meningitis, shock and death

Pathogenesis

The most studied strain of anthrax is the aimes strain

Consists of one chromosome and 2 plasmids (pX01 and pX02) that carry genes that encode the virulence factors.

Px01- protective antigen (PA)

Px02: -edema factor (EF)-Lethal factor (LF)-poly-D-glutamic acid capsule

Pathogenesis

Capsule: protects the bacteria in the vagitative state against phagocytosis.

PA: binds to the cell host's cell receptor and help AF And EF enter the cell by endocytosis.

EF: is an adenylate cyclase that affects all cells 1- converting ATP to cAMP2- cAmp concentration increases 3-water imbalance occurs 4-edema

pathogenesis

LF: metalloprotease that affects only macrophages-cleaves most activated protein kinase kinases MAPKKS and trigger an apoptotic pathway to lyses the cells

LF+PAnecrosis

EF+PAedema

EF+LFinactive

EF+LF+PAedema+ necrosis

Pathogenesis mechanism

The Fight against anthrax

vaccines and antitoxins target the protective antigen PA

Vaccines:-first-generation:(+) contained a culture from an attenuated strain (-) difficult to manufacture and has side effects-second-generation(+) contained nontoxic recombinant rPA molecule(-) slow and expensive

New breakthrough

Researchers at university of taxis medical branch

rapid immunization of large, at-risk populations after potential exposure to anthrax- vaccine applied to mucosal surfaces to replace intramuscular vaccination-There study examined the use of soybean oil-and-water nanoemulsions (NEs) as a mucosal adjuvant for an rPA vaccine to replace the aluminum adjuvant of the traditional vaccines

Results

NE immunization was effective in inducing both serum anti-PA immunoglobulin IgA and IgG antibodies

high titers of lethal-toxin-neutralizing serum antibodies

sGuinea pigs nasally immunized with rPA-NE vaccine were protected against an intradermal challenge with 1,000 times the 50% lethal dose

a needle-free anthrax vaccine requiring fewer doses and having fewer side effects than the traditional vailable human vaccine

results

Conclusion

although the rate of natural anthrax infection has declined, this topic remains a controversial one because of its relation to bio terrorism

this breakthrough is an excellent starting point and scientists should introduce the vaccine to humans and test it's success rate. In the future, research must focus on finding an antitoxin that can effectively cure vaccinated and non vaccinated humans after infection or exposure to the spores.

the fight against anthrax and all other biological weapons should be taken more seriously in the future to prevent any surprising attack from taking place.

Reference

Baillie, L. 2001. The development of new vaccines against anthrax. J. Appl. Microbiol. 9:609613.Bailey-Smith, K., Todd, S.J., Southworth, T.W., Proctor, J., Moir, A., 2005. The ExsA protein of Bacillus cereus is required for assembly of coat and exosporium onto the spore surface. J. Bacteriol. 187, 38003806.Franz D. 2009. Preparedness for an anthrax attack. Molecular aspects of medicine 30:503-510.Friedlander A. M. 1986. Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. Journal of Biological Chemistry 261:7123.Shadonly S., T. Smith. 2008. Anthrax. Journal of the American Veterinary Medical Association 233:63-72.