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The school of CAMIDRCS

(coalition against mysticism in defence of reason commonsense and science)

Month

August 2015

A conspectus history of biological warfare

According to recorded history and available sources the use of pathogenic and lethal organisms and their derivatives goes back to the BC’s we can mention Hannibals ‘snake on the ship‘ scenario,the use of dead animals to contaminate wells,the dipping of arrows in decomposing bodies.

In the 14th century AD the Black death due to Yersinia pestis which swept through europe, the near east and north africa can be pointed out as the greatest public health disaster in recorded history according to Gabriele de Mussi’s memoir which was published in latin but translated into english he stated that the epidemic which caused europe to lose more than its quarter population was caused by the Mongol armies hurdling of plague-infected cadavers into the besieged crimean city of Caffa.In 1343 the mongols under Janibeg beseiged Caffa and the Italian onclave in Tana following a conflict between Italians and muslim Tana.

This seige lasted till february 1344 the seige was lifted by Italian relief forces by neutralizing 15,000 mongol soldiers but Janibeg renewed the seige in 1345 which was again lifted after one year. The Italians blocked mongel ports,forcing Janibeg to negotiate and in 1347 the Italians were allowed to restablish their colony at Tana.

The fact that biological weapons are disasterously effective  is because of their subtility in the use of bioterrorism and biowarfare the list of pathogens include Yersinia pestis,Vibrio cholera, Yellow fever virus,Brucella species,ricin,mycotoxins Bacillus anthracis,ebola, Francisella tularensis,Coxiella burnetii,variola, monkey pox virus,Staphylococcal enterotoxin B,and botulinum toxin are among the few.

These natural evolved or artifically synthesised pathogens can be delivered through air as aerosols loaded on misiles,inserted into food and water supplies and detecting them is extremely challenging until they start causing devastations.The hall mark of  bioterrorism is mass casualities because that’s their intended purpose therefore, a health worker on the scene of mass epidemic must suspect an outbreak or a foul play.

As human ingeniuosity progresses the types of pathogens deployed by mad nations also is getting sophisticated not to mention what evolution adds on top of that when man’s raging pugnacity is aided with modern technology the effects are catastrophic.During the first world war the Nazis developed anthrax,glanders,cholera, and plant pathogens to use them against the USSR in which they caused glanders.

Eventhough, the Geneva protocol was signed in 1925 by 108 countries prohibiting chemical and biological agents during world war II the Japanese operated a secret facility called Unit 731  in Manchuria which sadistically experimented on prisoners they exposed 3000 people to pathogenic and lethal agents.

The United states,Britian,USSR,Iraq also had their own research programs.The biological and toxin weapons convention was signed by the USSR,US,UK and others banning development,production and stock pilling of microbes and their poisonous products in 1972 which had more ‘teeth’ than the 1925 protocol which in 1996 137 nations signed the treaty.

But surprisingly the Soviet union continued its research and production of offensive biological weapons program called Biopreparat in 1971 an accident occured while on going test on the Isand of Vozrozhdeniye causing an epidemic of small pox.

According to the US congress office of technology assesement 8 countries were reported to have undeclared offensive biological warfare programs in 1995 China,Iran,Iraq,Isreal,Libya,North korea,Syria and Taiwan.

It is possible to write an entire book on bioterrorism and biological warfare how they are manufactured,delivered and how to protect ourselves from these silent killers this article is just an epitome of this very voluminous subject of microbiology which i collected from various sources.

A Discourse on manual erythrocyte and Leucocyte count.

1when it comes to cell counting in the clinical setting inorder to aid the diagnosis of ailments it will be proper to mention its history how it was developed before we talk about how it is used.The haemocytometer as it is called is a piece of thick glass with ruled areas to count the number of various peripheral blood cells like erthrocytes (red blood cells),leucocytes (white blood cells),thrombocytes (platelets) which was develped by a french anatomist and histologist  Dr.Loius-charles Malassez (1842-1909) see figure 1 who studied medicine in paris and in 1875 attained the chair of anatomy at  college of france and was a member of the academy of medicine.

The fungus genus of malassezia and epithelial cell rests of malassez (ERM) in the field of mycology and dentistry hold his name as eponyms.Haemopoiesis is a term used to define blood cell production by the body haemopoietic stem cells are capable of differentiating into the various peripheral blood cells we sees on a blood film.

Generally haemopoietic stem cells differentiate into five blast cells (nucleated precursor cells) and these are:-1-proerythroblasts which form erthytrocytes.

2-Myeloblasts which form mature neutrophils,eosinophils,and basophils.

3-Monoblasts which form mature monocytes.

4-Lymphoblasts which form mature lymphocytes.

5-Megakaryoblasts which form mature platelets.

Haemopoiesis takes place  in the red bone marrow which is found in the epiphyses or ends of long bones such as the humerus and femur.In addition to the sternum,ribs,cranial bones,the vertebrae and the pelvis.Certain haemopoietic growth factors stimulate differentiation and proliferation of certain proginator cells for instance Thrombopoietin is a hormone that stimulate  platelet formation.

Colony-stimulating factor  and interleukins are cytokines which stimulate white blood cell or leucocyte formation.The counting chamber called Neubaeur counting chamber is a thick glass with 4 protruding shoulders and two counting ruled ares with 3mm2 area with a depth of 0.1 mm.

The ruled area is where the RBC,WBC and platelet are counted the area of 1,2,3,and 4 for wbc and the central A,B,C,and D for rbc and platelet count. see figure 22

RBC  count

when counting rbc per mm3 we use a tomma pippette with red bead in it with a mark 101.

Diluting fluid

formal citrate1%

Formula

Total rbc counted x 50 x 200

WBC count

when counting wbc we use the white bead tomma pippette with 11 mark.

Diluting fluid

acetic acid 2%

Formula

Total wbc cells counted x 2.5x 20

Reference ranges source:- Oxford medical dictionary

RBC:-4.5×6.5×10 12/L or 4,500,000-6,500,000 /uL

WBC:-4.0×11.0x10 9/L or 4,000-11,000/uL

neutrophil:-2.0 – 7.5×10 9/L or 2000-7000/uL

Lymphoctes:-1.3-3.5 x 10 9/L or 1,300-3,500/uL

eosinophils:- 0-0.44 x10 9/L or 0-440 /uL

Basophils:-0 – 0.10 9/L or 0-100 /uL

monocytes 0.2-0.8 X 10 9 /L or 200 – 800 /uL

What is haemoglobin ? and How it is measured ?

1

The haemoglobin molecule was shown to transport oxygen as  oxyhaemoglobin by Fredrich L.hunefeld [1799-1882] see figure 2 in 1840 he was a member of German biochemistry association while viewing the  blood of earth worm eventhough the molecular mass was determined earlier by J.F Engelhard in 1825  see figure 4.3

Haemoglobin is a molecule made up of two alpha and two beta molecules held together with 4 haem molecules. The life span of  a haem molecule is as the same as the RBC 120 days where the concentration of haemoglobin is 14-16 grams per 100ml and the iron content of each haemoglobin molecule is 3.4 nanograms.

At the end of the life span of Rbc macrophages in spleen,liver or red bone marrow phagocytize rbcs  and the globulin part is broken down to aminoacids which later on can be used to synthesize other proteins and from the haem part the iron is removed attaches to plasma protein transferrin and when reaching the liver cells,muscle fibers iron detaches from transferrin and sticks to storage proteins called ferritin and haemosiderin.

Later on iron is released from storage proteins and reattaches to transferrin and is transported to the bone marrow where it is taken up by rbc [red blood cells] precursors for utilization of new haemoglobin molecules.

The Haem part has non-iron parts which is turned into biliverdin which is a green pigment then this molecule is turned into bilirubin within the liver,passed to bile as conjugated bilirubin in the process of the hepatocyte by the action of the enzyme  UDP-glucuronyl transferase which  conjugates it with glucuronyl acid which makes bilirubin water soluble.

Bile then passes to the small intestine where it  is degraded by bacteria into colorless water soluble urobilinogen.Urobilinogen further oxidize to compounds called urobilin and stercobilin.stercobilins are  excreted via feces where as some urobilinogen is absorbed to blood and converted to urobilin and excreted via urine which gives urine its

color.2

There are two types of haemoglobin and 4 types of Haem [Haem a-C49H56O6N4Fe,Haem b-C34H32O4N4Fe,Haem c-C34H36O4N4Fe,Haem o-C49H58O5N4Fe] as represented via molecular configuration.

1-Haemoglobin A- is main adult haemoglobin with 2 alpha and 2 beta chains.

2-Haemoglobin F- a type of haemoglobin present intrauterine existence and at birth with 2 alpha and 2 gamma chains.

Haemoglobin may combine with other chemicals in a life threatening conditions like for instance.

1-carboxyhaemoglobin:- combination of carbonmonoxide and haemoglobin.

2-Methaemoglobin:-combination of sulphonamides,benzocaine,amylnitrite,chloroquine,dapsone,nitrites,nitriglycerin,nitropruside,phenacentin and others in which iron is oxidized from ferrous Fe 2+ to ferric Fe 3+ state.4

Measurement methods of Haemoglobin.

1-Acid haematin method / Sahli method.

is a visual technique of haemoglobin estimation named after its developer Hermann Sahli [1856-1933] see figure 3 where haemoglobin converted to acid haematin by H2SO4 sulphuric acid and the color produced is matched to a comparator see figure 5.9

Procedure

1-Fill the graduated tube upto the mark 20 with 0.1 N Hcl.

2-Fill the haemoglobin pippette exactly upto 20 mm3.

3-wipe off the blood outside pippette with gauze.

4-empty the pippette into the acid in the tube gently.

5-Mix the acid haematin solution in the tube  and allow to stand for 5-10 minutes.

6-place tube in sahli comparator.

7-start adding 0.1 N Hcl or distilled water drop by drop till the color matches that of the standard.

8-Read the volume of solution in the graduated tube and express as grams 100ml or as percentage.

Other methods of estimation like cyanmethaemoglobin method,Alkali method,sheard-Sanford oxyheamoglobin method,sodium lauryl sulphate method, also exist according to Ramnik soods textbook 2006 edition.

What is Chikungunya virus and chikungunya disease ?

220px-Aedes_aegypti_biting_humanThe name chikungunya comes from an african word and kimakonde language translated as ” that which bends up” because of the posture patients show while in the process of the malady.The origin of the virus is traced to south-east africa transmitted by Aedes aegypti and Aedes albopictus mosquitoes while being infected themselves in the first place they infect humans. The virus eventhough originated in south-east africa out breaks manifested in Asia,europe, and the Americas specifically  according to the WHO countries infected in Asia include:-Cambodia,east timor,India,Indonesia,Laos,Malaysia,Maldives,Mayanmar,Pakistan,Philipines,Reunion,Seychelles,Singapore,Taiwan,Thailand and vietnam.

Africa:-Benin,Burundi,Cameroon,Central african republic,Comoros,Congo(DRC),Equatorial Guinea,Kenya,Madagascar,Malawi,Mauritius,Mayotte,Nigeria,Senegal,South africa,Sudan,Tanzania,Uganda,and Zimbabwe.

The Americas:-US and the Carribean.

Europe:-Italy in 2007 due to immigrants.

The symptoms of this disease include Muscle pain,joint pain,nausea,fever/pyrexia 40’c,skin rash 4-7 days after the infection specific treatment doesnot exist except the management of symptoms reducing pain and fever via Ibuprofen and Non-steriodal anti-inflammatory drugs to suppress the immune symptoms eventhough they may be counter productive at this time of infection.The disease has a very low rate of fatality 21 deaths in 5,037 infection according to WHO.

People with senility,infants and patients with underlying diseases are particularly venerable.Caution and prudence for travellers to the area of infection include use of repellents,use of bednets,clothes which cover up all exposed skin using long sleeves,hats,long socks etc…Further more,Chikungunya prevention and prophylaxis depends  mostly in controlling  the Mosquitoes themselves by using insecticides according to proper guidance of WHO (pesticide evaluation scheme),disposal of water-storage objects like old tyres,flower vessels,discarded plastic containers which will deprive the mosquitoes a place of development and growth.

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