A SEMINAR ON THE USE OF BACTERIA AS BOLOGICAL WEAPONS

SUMMARY

Bioweapons is a planned and deliberate use of pathogenic strains of microorganisms such as bacteria, viruses or their toxins to spread life-threatening diseases on a mass scale in order to devastate the population of an area. Bacterial weapons include Bacillus Anthracis, Yersinia pestis, Coxiella burnetti and so on. Bacillus Anthracis is a gram-positive aerobic bacillus, with both a spore forming and vegetative form. Anthrax spores can survive in their dormant state, typically in soil, for decades until they come with a host through cutaneous breaks, inhalation or ingestion. Characteristic of bioweapons include easy to attack and deliver, ability to attack in a reproducible and consistent manner, etc. Signs and Symptoms include ; Vomiting, Headache, Fever and Chill. It can be diagnosed in a laboratory through blood or cerebrospinal fluid. Preventive measures include prophylactic vaccination and childhood immunization, etc. Public enlightenment should be  encourage by the government to minimize the spread of anthracis.

TABLE OF CONTENTS

Tittle  page                                                                                                                  i                                                                                                                                                                        

Certification                                                                                                                ii

Aknowledgement                                                                                                        iii

Summary                                                                                                                     iv    

Table of Content                                                                                                           v

  Introduction                                                                                                           1

 Biological warfare and Biological weapons                                                        2

2.1 Characteristics of  Biologcal weapons                                                                   3

2.2 Candidate Biologcal weapons                                                                                3

3.0 Bacillus Anthracis                                                                                                  4

3.1 Biology and Growth of Bacillus anthracis                                                           5           

3.  Infection and Pathogenesis of B. anthracis                                                              5

3.3 Pictoral image of Pathogenicity of Anthrax                                                         7

3.4 Sign and Symptoms of B.anthracis                                                                       8

3.5 Gastrointestinal and Weaponized Anthrax                                                            8

3.6 Vaccination, Prophylaxis, Treatment, and Diagnosis of  B.anthracis                  10

4.0 Cutaneous and  Inhalational Anthrax                                                                      11

4.1 Micoscopic View of B.anthracis                                                                          13  

4.2 Cutaneous Lession of B. anthracis                                                                       14

5.0 Conclusion and Recommendation                                                                        15

References                                                                                                                   16

                                                                           

1.0 INTRODUCTION

Bioweapons is a planned and deliberate use of pathogenic strains of microorganisms such as bacteria, viruses or their toxins to spread life-threatening diseases on a mass scale in order to devastate the population of an area. The September 11, 2001 hijacking of four U.S. airliners and the subsequent striking of three of them into the two towers of the World Trade Center and the Pentagon was the worst mass casualty terrorist attack (Dudley,  2010).

       In U.S. history. Approximately 3,000 people from over 80 countries lost their lives at the World Trade Center in New York. By addressing the broad spectrum of biological threats through a rather heterogeneous system of legally- or politically-binding agreements and practices among diverse stakeholders, the Biological Weapons Convention (BWC) and the United Nations Security Council Resolution 1540 (UNSCR 1540) effectively belong in a unique category of international regimes aimed at biological risk management (Parkins  and Danskn , 2011). A bioterrorist attack releases viruses, bacteria, or other germs to cause illness or death. These biological agents are typically found in nature. But they can sometimes be made more harmful by increasing their ability to cause or spread disease, or to resist medical treatment. These bacterial weapons is considered a potential biothreat agent not only due to potential threats to human health and also because of agricultural concerns and ecotoxicological hazards (Kumar et al, 2010).

The terrorist use of airliners as missiles, the United States was faced with a series of anthrax attacks delivered to victims and target offices through the U.S. mail system. There was a series of anthrax attacks during 2001 via U.S. Postal Service fulfilled the warnings of those who had warned of the inevitability of future bioterrorist events. The anthrax attacks of 2001 in the United States validated the previous warnings by some experts concerning bioterrorism that it was not a question of “if ”; it was a question of “when.” Now such bioterrorism is a historical fact, not just a prediction of the future (Christopher  et al, 1997). Terrorists may find biological agents to be an attractive alternative  to conventional weapons because of their relatively low costs, their relative accessibility, and the relative ease in which they could be produced, delivered, and avoids detection. Their use, or even threatened use, is potentially capable of producing widespread social disruption. Bacterial agents such as Bacillus Anthracis, Yersinia pestis, Brucella Abortus and Coxiella burnetti could be used as agents for bioterrorism. Research into the use of Bacillus anthracis as a bioweapon is at least 80 years old and several nations are believed to have weaponized anthrax (fowler and Shafazand,2011). 

2.0 Biological Warfare and Biological Weapons

Biological warfare is the use of biological agents to incapacitate or kill a military or civilian population in an act of war or terrorism. Biological weapons have been used against targets in  the United States,  Historically, Biological Warfare (BW) agents of concern have included a relatively select group of pathogens and toxins, referred to as traditional BW agents. Traditional biological warfare agents are all naturally occurring organisms or their toxic products. From the perspective of biological warfare scientists, traditional BW agents have serendipitously evolved a select group of traits: toxicity, stability, and ease of production. However, until recent decades, the serendipity that aided researchers in choosing select organisms also limited BW applications to the characteristics of available agents. For example, environmental stability, infectious dose, time to effect, clinical progression, and lethality are all properties intrinsic to candidate traditional agents that may limit their utility for biological warfare.Technologies developed across multiple disciplines and as a result there is a potential to revolutionize biological warfare by facilitating an entirely new class of fully engineered agents referred to as Advanced Biological Warfare (ABW) agents (Inglesby et al, 2002).

2.1 Characteristics of Biological Weapons

Biological weapons are organisms or toxins that are:

Easy to produce and deliver.

Safe for use by the offensive soldiers.

Able to incapacitate or kill individual attack in a reproducible and consistent manner (Christopher, et al., 1997).

Although biological weapons are potentially useful in hands of conventional military forces, the greatest likehood of biological weapons use is probably by terrorist groups. This is in part due to the availability and low of producing and propagating many of the organisms useful for biological warfare. Biological weapons are accessible in nearly every government and well-'financed private organization.

2.2  Candidate Biological Weapons

Virtually all pathogenic bacteria  are potentially useful for biological warfare, and several of the most likely candidate organisms are relatively simple to grow and seminate. The most commonly bacterial weapon is Bacillus anthracis, the casual agent of anthrax(James et al, 2003).

Other  bacterial weapons include;

Yersinia pestis, (The organism responsible for plague) 

2.Brucella abortus (fever and bacteremia; brucellosis)

3. Clostridium botulinum (Respomsible for botulism)

4. Francisella tularensis

5. Coxiella burnetti

6. Burkholderia pseudomallei and Burkholderia mallei

3.0 Bacillus anthracis

 Bacillus anthracis is a gram-positive aerobic bacillus, with both a spore forming and vegetative form. Anthrax spores can survive in their dormant state, typically in soil, for decades until they come into contact with a host through cutaneous breaks, inhalation or ingestion. Anthrax is a zoonotic disease (Zoonotic diseases are diseases that can be transmitted from animals to humans and from humans to animals) that is transmissible through handling or consumption of contaminated animal products. The etiologic agent of anthrax, Bacillus anthracis, is a spore forming gram-positive bacillus. Areas currently listed as high risk are South and Central America, Southern and Eastern Europe, Asia, Africa, the Caribbean, and the Middle East. In late 2001, a deliberate dissemination of Bacillus anthracisspores via letters sent through the U.S. Postal Service resulted in 12cases of cutaneous anthrax and 11 cases of inhalation anthrax Humans can become infected with B. anthracisby handling products or consuming under cooked meat from infected animals. Infection may also result from inhalation of B.anthracis spores from contaminated animal products such as wool or the intentional release of spores during a bioterrorist attack. Cutaneous infections occurs when the bacterium or spore enters a cut or abrasion on the skin, such as when handling contaminated wool, hides, leather or hair products (especially goat hair) from infected animals. Approximately 20% of untreated cases of cutaneous anthrax result in death. Cutaneous anthrax usually appears in the form of a localized, painless, central black eschar with surrounding oedema  (Christopher et al., 1997).

3.1 Biology and Growth of Bacillus Anthracis

Bacillus anthracis is a ubiquitous saprophytic soil inhabitant. It grows as an aerobic gram-positive rod, 1 μm in diameter and 3-4 μm in length. As with other species of the genus Bacillus, B. anthracis produces endospores resistant to and drying. Endospore formation enhance the ability to disseminate B. anthracis in aerosols endospores are sometimes recovered from contaminated animal products such as hides and fur. Growth on blood agar results in large colonies with a characteristic “ground glass appearance. Strains having a poly-D-glutamic acid capsule are resistant to phagocytosis (Ryan, 2016).

3.2   Infection and Pathogenesis

Bacillus anthracis endospores are the standard means of acquiring anthrax. The disease usually affects domestic animals especially ungulates—cow, sheep, and goats. The number of infections in animals, although considerable is not known. The animals acquire the disease from plants or soil in pasture. In humans and animals, there are three forms of disease Cutaneous anthrax, is contracted when abraded skin is contaminated by B. anthracis endospores  Gastrointestinal, anthrax is contracted from consumption of endospore-contaminated plants or anthrax-infected carcasses Cutaneous anthrax cases are rare in the United States. Human gastrointestinal anthrax is rarely seen. Pulmonary anthracis is contracted when the endospores are inhaled. Inhalation of the endospores or the live bacteria results in pulmonary and inhalations characterized by pulmonary and cerebral hemorrhage. Untreated pulmonary anthrax infections have a mortality rate of nearly 100%. Pulmonary anthrax are even in agricultural workers, extremely rare. The last naturally acquired pulmonary anthrax case in the United States occurred in 1976. However, several cases of pulmonary anthrax were identified in 2001 due to bioterrorism event (Christopher et al., 1997).

Pathogenesis results from inhalation of 8000-50,000 endospores of an encapsulated toxigenic strain. Pathogenic Bellusic anthracis produces three proteins protective antigen (PA), lethal factor (LF), and edema factor (EF). PA and LF form lethal toxin. PA is the cell-binding, Bio-component of these AB-type toxins. EF cause edema, and LF cause cell death. Growth of B. Anthracis in the lymphinodes and lymphatic tissues graining the lungs to edema and cell death, culminating in tissue destruction, shock, and death.(Mayer et al., 2001).

3.3         Pictorial Image of pathogenicity of Anthrax

Sign and Symptoms

Clinical symptoms can start with sore throat, fever, Vomiting, Weakness, Headache, Abdominal pain, Dyspnea, muscle aches. After several days, symptoms include difficulty in breathing, followed systemic shock. Fatality rates approach 90% even when exposure is recognized and treatment is started, and can be nearly 100% in cases for which treatment is not started until after one onset of symptoms (Mayer, et al., 2001)

3.5.0 Gastrointestinal and weaponized Anthrax

Gastrointestinal Anthrax is rare but serious bacterial disease. A person can acquire gastrointestinal anthrax from eating contaminated meat from infected animals  or their spores and is characterized by an acute inflammation of the intestinal tract. Many factors secreted by monocytes and macrophages contribute to chronic inflammation . Initial signs of nausea, loss of appetite, vomiting, and fever are followed by abdominal pain, vomiting of blood, and severe diarrhea. The mortality rate is difficult to determine for gastro intestinal anthrax but is estimated to be 25%-60%.  However, weaponised anthrax is a bological agents or germs that can sicken or kill people, livestock or crops (Jocobsen et al., 2011).

3.5.1 Gastrointestnal Anthrax

 Gastrointestnal Anthrax  occurs following depositon and subsequent germinaton of spores in the upper or lower gastrointestinal tract. The former results in the oral pharyngeal form of disease. An oral or esophagical ulcer leads to developments of regional lymphadenophathy, edema, and sepsis. The latter resides in primary intestinal lessons occurring predominantly in the terminal ileum or aecum, to preventing initially with nausea, vomiting, and malase and progressing rapidly to bloody diarhea, acute abdomen or sepals. Massive as site has occurred in some cases of gastrointestinal tract anthrax. Advance infection may appear similar to the sepals syndrome occurring in either inhalational or cutaneous anthrax. Some authors suggest that aggressive medical intervention would be recommended for inhalational anthrax may reduce mortality, although given the difficulty of early diagnoss, mortality almost inevitably would be high (Shea and Golfron, 2004).

3.5.2 Weaponized Anthrax

The term weaponized is applied to strains and preparations of B. anthracis, usually endospore form, that exhibit properties that enhance dissemination and use as biological weapons. Such strains and preparations 'were developed in several countries in the post-World War II era, but overt development of biological weapons was halted by international treaty in 1972. The physical characteristics of the weaponized anthrax preparations typically include a small particle size, usually interspersed with a very fine particulate agent such as talc. This small-particle, powdery forth ensures that the endospores will spread easily by air currents. Thus, opening an envelope containing endospores or releasing the powder-endospore mixture into a ventilation system or other air current has the potential to contaminate surrounding areas and personnel (Shea and Golfron, 2004).

A weaponized form of anthrax was used in a series of bioterrorism attacks in the United States in 2001. These incidents were carried out by mailing envelopes or packages containing weaponized anthrax endospores. The attacks were apparently directed at the news media (Florida) and the government (Washington, DC area). A third focus of attack, the Pennsylvania-New York area, had no defined single target but disrupted mail service in the Northeast; some of the anthrax-contaminated mail facilities were still not in use 2 years later. In all, there were 22 anthrax infections. Eleven were cutaneous anthrax. Of 11 cases of inhalation anthrax, 5 cases resulted in death. The bioterrorists were never identified(Swartz, 2001)

The incidents in the United States were not the first or the most serious anthrax biological weapons infections. In a previous incident, B. anthracis spores were inadvertently released into the atmosphere from a biological weapons facility in Sverdlovsk, Russia, in 1979. Less than 1 g of endospores was released, and everyone in the area surrounding the facility was immunized and given prophylactic antibiotic therapy as soon as the first anthrax case was diagnosed. However, 77 individual outside the facility contracted pulmonary anthrax and 66 died (Swartz., 2001).

Vaccination, Prophylaxis, Treatment, and Diagnosis.

Vaccinaton is the admnistration of antegenic material to stimulate an individual immune system  to develop an adaptive immunity to a pathogene. Vaccines can prevent ameliovate infectous disesase. When a suffciently large percentage of a population has been vaccined, herd immunity results. The effectivenessof vaccinaton has been  widely studied and verified. Vaccinaton is the most effective method of preventing infectous disease (Liesegang and Thomas, 2009).

    Prophylaxis is a measures  taken to mantain health and prevent the spread of disease as opposed to disease  treatment. Just as health comprise  a variety of  physical and mental states, so do disease and  disability whch are affected by environmental factors, genetc predisposition, disease agent and life style  choices. Health disease and  disabilty are dynamic processes which begin before indvduals realize  they are affected. Disease prevention  relies on anticpatory actions that can be categorized as Primal, Primary, Secondary and Tertiary prevention (Liesegang and Thomas, 2009)

   Treatment is the medical care given to a patient for an illness or injury, it is the act or method of handling or dealing  with someone or something. it also involves putting a particular substance onto or into it in order to clean it, to protect it or to give it special properties while diagnosis is the process of determining which disease or condition explains a person Symptom and Sign. It is most often reffered to as diagnosis with the medical context being implicit. The mformaton  required for diagnosis is typically collected from a history and physical examnation of the person seeking  medical care often  one or  more  diagnostic procedures, such as diagnostic test are also done during the process (Perkins and Danskin, 2011)

. 3.6.1 Vaccinatiion

Vaccination for anthrax has thus far been restricted to individuals who are considered at risk. This includes agricultural animal workers and military personnel. The current vaccine, called anthrax vaccine adsorbed (AVA), is prepared from a cell-free B. anthracis culture filtrate. The 135 anthrax vaccine, an inactivated cell free products was licensed n 1970 and is produced by flopord corp, laming, mich (formerly called the machigon halogni products institutes). The vaccine is licensed to be given in a 4 dose sellers and has recently been mandated for all US military active and reserve duty personnel. 51 the vaccine is made from the cell free filtrate of a non encapsulated strain of B. anthracis the primary antigen responsible for inducing immunity been shawn in 1 small placenotor controlled human trial to be efficacious against cutaneous anthrax. 53 as of march 1, 1999, approximately 590,000 doses of anthrax vaccine has been administered to US armed forces (Gary strawder, Department of Defence, falls church V9, Oral communication April (1999); no serious adverse events have been causally related (Mles Braun, Food and Drug administration, Rockvlle, innoculation with this vaccine at 0 and 2 weeks was completely protective against an aerosol challenge at 8 and 18 weeks  and 85% effective at 100 weeks (Inglesby et al., 2002).

3.6.2 Treatment

Treatment of B. anthracis infection, which seems to have a minimum incubation time of about 8 days, is usually done with antibiotics. Ciprofloxacin, a broad-spectrum quinolone antibiotic, is used against strains that are penicillin-resistant, including; many laboratory and biological weapons strains. Ciprofloxacin is also used as a prophylactic measure to treat potentially exposed individuals(Prescott and Dawling, 2013).

3.6.3 Diagnosis

Rapid diagnostic tests are available to detect microbial endospores. However, positive identification of B. anthracis relies on culture techniques and direct observation of either infected tissues or cultured organisms. The characteristic ground-glass appearance on blood agar, coupled with the isolation of gram-positive endospore-forming rods growing in extended chains, is presumptive evidence for B. anthracis.(Prescott and Dawling,2013).

 4.0 Cutaneous  and Inhalational Anthrax.

 Cutaneous anthrax is the process where by infections occurs when the bacterium or spore enters a cut or abrasion on the skin, such as when handling contaminated wool, hides, leather or hair products (especially goat hair) from infected animals. Approximately 20% of untreated  cases of cutaneous anthrax result in death. Cutaneous anthrax usually appears in the form of a localized, painless, central black eschar with surrounding oedema ( Jocobsen et al, 2011)

     Inhalational anthrax is a form of anthrax results from inhaling B. anthracis spores and is most likely following an intentional aerosol release of  B. anthracis. Fever, malaise, and fatigue maybe present initially, sometimes in association with a nonproductive cough and mild chestdiscomfort. Severe respiratory distress with dyspnea (labored breathing),diaphoresis(perspiration), stridor (high-pitched whistling respiration), and cyanosis (bluish skin color) B. anthracisis considered a potential biological warfare threat agent (Sridhar and Chandrashekhar, 1996)

. 4.0.1  Cutaneous anthrax:

 The blackened lesion on the foreman of a patient, about 2cm in diameter, results from tissue necrosis. Cutaneous anthrax, even when untreated, usually is a localized non lethal infection. There are five doses to suggest the personalitiy of a prolonged laboratory period in cutaneous anthrax.  in anthrax, cutaneous cases surround only as days after the spore germinaton in skin disease, toxin production results in local edema, An initially prioriotic macule or papule enlarges into a wound illness by the second day. Subsequently, 1-to-3mm vessicles may appear, which discharged clear serogunoid flud containing numerous organisms or gram stain. Development of a painless, depressed black Escher follows, often associated with extensive local edema. The Escher dries, loosens and fell off in the next 1-to-2 weeks, most often leaving no permanent scarlymphagitis and painful lymphadenopathy can occur with assaocated system symptoms. Although antibiotic therapy does not appear change the course of Escher formation and healing.  It does increase the likelihood of systemic disease without iantbotc therapy, the mortalitiy rate has been reported to be as high as 20% with antibiotics, death due to cutaneous anthrax is rare (Mayer et al, 2001).

 4.0.2  Inhalational anthrax:  

Inhalational anthrax is considered to be the most deadly forms of anthrax. Infection usually develops within a week after exposure, but it can take up to 2 months without treatments, only about 10-15% of patients with inhalational anthrax survive. However,  with aggressive treatment, about 55% of patients survive. When a person breathes n anthrax spores, they can develop inhalation anthrax. People who work in places such as wool mills, slaughterhouses, and tanneries may breathe in the spores when working with infected animals. inhalation anthrax starts primarily in the lumph node in the chest before spreading throughout the rest of the body, ultimately causing severe breathing problems and shock The fixed human brains shows hemorrhagic meningitis (dark coloration) due to a fatal case of inhalation (Mayer et al., 2001).

4.1 Microscopic view of Bacillus Anthrax

:Source:  www.anthrax.osd.mll

Microscopic view of  B. anthracis

Fgure 4.11

4.2  Cutaneous lesions of B. Anthrax

Source: www.anthrax.osd.mll 

Cutaneous lession of B. anthracis

Figure  4.21

5.0 Conclusion and Recomendations

5.0.1 Conclusions

Because of the increased threat of terrorism and the risk posed by various microorganisms as biological weapons needs to be evaluated  and the historical development and use of biological agents should be better understood. Biological warfare agents’ in particular bacterial bio warfare agents may be more potent than conventional and chemical weapons. During the past century, production of such weapons. In addition, genetic engineering holds perhaps the most dangerous potential of biothreat. Ease of production and the broad availability of biological agents have led to a further spread of biological weapons and an increased desire among developing countries to have them.

5.0.2 Recommendations

It is  recommended that  an assessment of the risks associated with the use of pathogenic microorganisms required in order to develop countermeasures to limit unexpected scenarios, characterized by mass destruction and to assure a ready response to them .

      Bioweapons also remain one of the legitimate and perhaps growing military and terrorist threat in the current world situations. However, Knowledge of inhalational anthrax is necessary for public health officials, as well as the health care providers who will be call on to care for casualties. 

  

References

Binkley, C,E., Cinti, S. Structure.D.M.,and  Acolletti, L.M. (2002). Bacillus anthracis as an agent of bioterrorism a review emphasizing surgical treatment, Anthracis of surgery. 236(1), 9-16.

Christopher GW, Cieslak TJ, Pavlin JA,and  Eitzen EM Jr (1997) Biological warfare.A historical perspective. JAMA 278: 412-418. 

 Christopher, L.G.W., Cieslak, L.T.J., Pavlin, J.A. , and Eitzen, E,M (1997). Biological warfare: a    historical perspective. Jama, 278(5), 412-417.

Colonel Jim A. Davis and Dr. Barry R. Schneider (2002) The Gathering Biological Warfare Storm. (2002 edition),Greenwood Publishing Group, Alabama, USA. 8:225-250

Dudley JP (2010) Review and Analysis of Reported Anthrax-Related Military Mail Security Incidents in Washington D.C. Metropolitan Area during March2005. J Bioterr Biodef 1: 101. 

Edgar J. DaSilva (1999) Biological warfare, bioterrorism, biodefence and the biological and toxin weapons convention. Electronic Journal of Biotechnology 2: 99-129. 

Fowler RA, and Shafazand S (2011) Anthrax Bioterrorism: Prevention, Diagnosis and Management Strategies. J Bioterr Biodef 2: 107.

Friedlander A,M., (2011) Anthrax clincal features, Pathogenesis and potential biologcal warfare threat. In: Remington J.S, Swartz. M.N, eds. Current clincal topic in infectous disease, (20), Maldan, Massachusetts: Blackwell Sciences, 2000-335-49

 http://biology.about.com/od/biotechnologycloning/a/biological-weapons.htm. 

http://www.bt.cdc.gov/bioterrorism/overview.asp. 

http://www.owr.de/go/owr/home/knowledge/biological_warfare_agents.xhtml.

Jacobsen S, Hugh-Jones ME, and Rosenberg BH (2011) The 2001 Attack Anthrax: Key Observations. J Bioterr Biodef S3: 001-11 

 Inglesby, T.V., O’toole, T., Henderson, D.A., Bartlett, J.G., Ascher, M.S., Eitzen, E., and  McDade, J. (2002). Anthrax as a biological weapon, 2002: updated recommendation for managemen. Jama, 287(17), 2236-2252

James BP, Theodore RP, and Jack AM (2003) Biotechnology: Impacton BiologicalWarfare and Biodefense. Biosecur Bioterror 3: 161-168. 

Kumar  NK, Reddy DSR, and Venkateswarlu P (2010) Application of Response Surface Methodology for Optimization of Chromium Biosorption from an Aqueous Solution onto Syzigium cumini (java) Seed Powder. J Microbial Biochem Technol 2: 20-27.

Lew, D. (1995). Bacillus anthracis (anthrax). Principle and practice of infectous disease. 4th ed. Churchill livingstone, 1985-1889. 

Liesogang and Thomas J (2009).`` Vercella zoster vrus vaccines:effective but concern lnger``. Canadan journal of ophthalmology 44(4):379-384.

Mayer, T,A., Horsoff Match., N., Murphy. C., Forls. T., Herpes. S., Pause, D. and  Hanflung.. D. (2001). Clinical presentation of inhalatonal anthrax following bioterrorism exposure; report of 2 surviving patients, Jumas, 286 (20) 249-2533.

Perkins D, and  Danskin K (2011) On the Front Line of Biodefense: The U.S. Department of Health and Human Services Support to International Biological Risk Management Regimes. J Bioterr Biodef 2: 111. 

Prasad, S.K. (2009). Biological Agents (Vol. 2). Discovery Publishing House.

Simon JD (2011) Why the Bioterrorism Skeptics are Wrong. J Bioterr Biodef S2: 001. 

Source:www.anthrax.osd,mll/pectoral image of pathogenicity of anthrax

 Stajner I (2009) Cloudiness and Breast Cancer. J Cancer Sci Ther 1: 34-40

Prescott, J.F., and  Dawlng, P.M (eds) 2013. Antimcrobal therapy on veterinary medicine. John wiley and sons.

Ryan, J, (2016). Bosecurty and bioterrorism contaning and preventing biologcal threats, Butterworth-Hennemann.

Shea, D.A., and  Golfron, F. (2004,may0. Small scale terrorist attacks using chemical and biological agents: An assessment of framework and preliminary comparisons. Library of Congress Washington D.C  Congressional  Research Service.

Swartz, M.N-162.N.(2001). Recogintion and management of anthrax-an update, new England journal of medicine, 345(22), 1626

. Sridhar SM, Chandrashekhar P (1996) Cutaneous anthrax with secondary 

          infection. Indian J Dermatol Venereol Leprol 57: 38-40. 

Wong G, Richardson JS, Pillet S, Schindle S, and Ennis J,  (2011) Evaluation.