images of Dry Tortugas National Park Photography by aircraft

 

images of Dry Tortugas National Park Photography by aircraft

 Dry Tortugas National Park beauty of the blue sea and golden sands


, About 70 miles to the west of the city of Key West in Florida, is one of the national parks charming on the planet
It Dry Tortugas National Park and world-renowned because of picturesque blue waters of the sea, and the golden sands,
The brightly colored coral reefs, and abundant marine life, shipwrecks and sunken treasures.


The park includes seven small islands area of ​​each island is estimated at 100 km, where they are accessed either through the small boats or seaplanes,
Park includes the largest building in the Western Hemisphere, which was built more than 16 million pieces of bricks,
The national park includes coastal castle contains 2000 of beautiful architectural brackets.
We travel together to this charming garden through these amazing images

images the beauty of nature in Portugal

images the beauty of nature in Portugal

Pictures on a well known Ik Kil in Mexico

Pictures on a well known Ik Kil in Mexico

(Ick all \ Ik Kil) is a well in the very magnificence and beauty, called the local population of this type of natural formations wonderful name (Synnott \ cenote), and well is located in the city of Piste in the center of Mexico's Yucatan Peninsula, and is well underground part of the (park "ick" archaeological) located next to the city (Chichen Itza \ Chichen Itza), which contains one of the pyramids of the ancient Mayan people that have been built for more than 500 years BC.

Located basin at a distance of 26 meters below the surface and a width of about 60-meter depth is water in it about the 40 meters, was drilled Mayan ancient ladders gradient in the rocks to facilitate access to this well and there are archeological excavations in large cover that area, so watch and visit thisLocation continues to impress visitors who stand amazed between the wonderful magic of nature with fragrant ancient history of the Mayan people.

The park "ick all" destination for tourists who visit the ancient pyramids located in the area where visitors are allowed to Asobho in well water groundwater was built a number of restaurants, shops and inns in the region has hosted the well one Champions Red Bull in diving as world events that attractenchanting beauty of nature to this wonderful well.

\

Know the images on the French city of Strasbourg

 

Know the images on the French city of Strasbourg

Strasbourg (French: Strasbourg) is a French city located in the east of the country with a population of 704,000 people, and including areas surrounding the city (Statistics 2007)
Strasbourg this small city was destroyed in the Middle Ages between the sixteenth, seventeenth and during religious conflicts and created new existing Bhltha with previous restoration of historic buildings.
Strasbourg is the capital of the Alsace region .. Which has long disputed sovereignty of Germany and France.
There by the European Parliament European Parliament
There is also the Council of Europe Council of Europe


Of Germany are close to a border town away from the German city 20 kilometers agentAnd away from the city Disney Iorb Games 15 kilometers Games the largest city in Europe ..
Passes by the Rhine River and proof and is location for Europe at a crossroadsClose to Frankfurt and Munich and Baden and Zurich, Interlaken and Zell am See headquarters between time to three hoursWhich is closer to her Paris as its capital.
Sons of the town who shot to prominence Madame Tussauds (her name Mary Krochwltz), which was born in the same city and grew upIn Paris and lived in London and created a wax museum, which embodied the wax statues of celebritiesIs a famous shrine in the British capital.
The Strasbourg second capital of the European Union after Brussels.
Praises a large mosque in Strasbourg alternative to the lounge where Muslims gather to pray and only accommodates 500 worshipersAnd the mosque will be a prominent architectural masterpiece filled with worshipers who are eager to end the business in 2008.
City landmarks and Dam Barrage Vauban prison
The advantages of the city and it is surrounded by attractive charm channels water from all sides .. And small aisles .. The old buildingsThey are similar to the Italian city of Venice and the Belgian city of Bruges and the Dutch city of Amsterdam in terms of waterwaysBut here there is only surrounded by channels from the four and is the smallest of those citiesAnd characterized by vehicle access to the main squares .. The tourist can walk around the city on foot smoothly and easily

I leave you with images

images Xiying Bridge in Taiwan ..

images Xiying Bridge in Taiwan ..

Bridge and footpath is the colors of the rainbow .. And located in Moukng County in Penghu in Taiwan ..
This bridge is characterized as reflecting neon lights on the surface of the water at night to form a fascinating spectrum ..

The bridge in the day:

Here in the night:

images urchin called 500 shares of thorns on dog attacked him!!

images urchin called 500 shares of thorns on dog attacked him!!

 

Counterattack 500 shares truck, was the dog attacked Guenvma share in Oklahoma City, but he fired a barrage of arrows struck her face and part of her body, according to a newspaper reported "the New York Daily News" American.

According to the source, the bitch "Bella Mae" aged 3 years, revolved around a pond in the backyard of a house مالكتها in Oklahoma when erased Guenvma. She added that the bitch was accompanied by other dogs, and although it did not approaching the Hedgehog, but he fired arrows at them, and the share of "Bella" is great.

The owner of the bitch said Alison Knowles, said the situation was terrifying, noting that despite the withdrawal of the majority of arrows they still find some of them in the face of "Bella". She believed that the hedgehog was drinking from a pond water when he launched his attack on the dogs. Quoted bitch to Animal Emergency Center where a doctor and two nurses took two hours to withdraw spinal arrows from her face and her mouth and lists.

Rapid flickering Chicarda tips to calm rapid heartbeat

Rapid flickering Chicarda tips to calm rapid heartbeat

This situation comes suddenly, and you do not understand something about the condition of your heart and all of a sudden become does go fast! The increasing number of pulses of 72 beats per minute to 120-180-200 in seconds pulse, breathing and may increase as well, and become a state of nausea with panic, and begin to sweat.The doctor says: You infected does go fast, and specifically, that you have accelerated heart paroxysmal, no doubt that once injury in this case for the first time you make a full examination, have you and your doctor to exclude injury quickly ventricle (a type of rapid heartbeat life threatening) and all diseases organic heart like thyroid disorders and pulmonary insufficiency. . . Etc.. That made you feel reassured.However, often come out atrium - a room in the heart receives blood from the veins and pumped into the ventricles - a bit of control, The atrium has a regular rhythm, but this rhythm can be faster than the normal three times.I know that the rapid flickering indicates increased heart rate faster than 100 beats per minute.
There are ways and means to reduce the heart rate, and below you will find methods to help you overcome the heart, in addition to tips on lifestyle for the prevention of heart:You calm:See this situation as a red flashing signal and says: stop what works, step back and tried to calm down. The rest is the best way to stop any bout.Vagal maneuver by taking a deep breath:The nerves Asambthawayh and nerves Albarasmbthawayh is regulating heart rate and strength of contractions (known as nerve Ha'ir) when the heart beats, the network will be Asambthawayh is dominant (a device that requests from the body mainly to accelerate), and what you should do is to shift control to the network nerves Albarasmbthawayh quieter, if affected vagal you begin the process of chemical rush of heart in the same way that trample on the brake the car to affect the speed.Another way to do this is to take a deep breath.Try to reach the carotid artery through rubbed:It is true that the right carotid artery massage is another way to vagal maneuver. Make sure that your doctor may show you the correct position and the right degree of pressure.You must be massaged at the confluence artery in the neck, under the jaw as far away as possible.Recall the reaction of the body to dive by immersing the face in ice water:When delve marine mammals in the coldest areas of water, Itbatye, heart rate automatically, and this natural way to keep the brain and heart animal, and can summon a reaction your body for diving in cold water so that fills a basin with iced water and immerse your face in it for a second or two.Stopped taking coffee, cola, tea and chocolate, slimming pills or stimulants in any form:The excessive intake of stimulants can expose you to the risk of fast heartbeat paroxysmal atrial fibrillation.Be sure to take care of the hypothalamus through proper diet and exercise:What is happening inside your head - specifically the hypothalamus - governs the heart - and this is why it is important to give the necessary attention to this part of the brain, through food الناسب and exercises to maintain stability and control of the future of the nervous system.The nervous system of the future is divided into two subgroups: the sympathetic nervous system future, which speeds up everything in the body except digestion, and the nervous system Albarasmbthawy.The tension and malnutrition and pollutants can make the hypothalamus loses control of the autonomic nervous system and allows the device to speed up the so-called sympathetic overload device future.You can help the hypothalamus in maintaining control.Eating a healthy diet and regular and reduced the dessert:If neglected some meals and then filled the stomach with candy or soft drinks, the pancreatic enzymes will increase to interest the amount of sugar large addressed, then increases insulin than normal and become a lack of sugar, and start glands adrenaline secretion of adrenaline to mobilize glycogen stores in the liver and then cause adrenaline a sudden increase in heart rate and a feeling of panic.You have to adapt the meals according to the table metabolismIt should people who have fast metabolism can eat foods rich in protein. The protein foods take longer to be digested and help prevent low blood sugar, when low blood sugar, it causes a process that we have discussed previously.You calmObserved a relationship between individuals who are characterized request perfection and who want height and repeat success and rapid heartbeat paroxysmal atrial fibrillation. Generally, these are people who have migraines, and for such, the mechanisms heart become exaggerated a lot, no overeating of chronic secretion of adrenaline, when exposed people to pressure nervous tremendous progress is made a surprise palpitations rapid and disappears irregular rhythm.How is compensation?You to follow a progressive relaxation program and dynamic exercise relax or learn how to imagine the serenity and calm, tranquility and security.Just eat yours from magnesiumThe magnesium protects cells, and for heart muscle cells, magnesium helps to offset the effect of calcium. When entering calcium into cells, it stimulates contractions of muscle within the same cell, and is magnesium necessary for enzymes that pumps calcium out of the cell, it causes constriction and relaxation in regular rhythm, it also makes the heart less susceptible to turbulence, and magnesium found in foods such as soy, nuts and peas, bran.Be careful on the high level of potassiumThe potassium is a mineral that helps to slow down the heart rate and uncontrollable muscle fibers, and no metal in some fruits and vegetables, and therefore obtain a sufficient proportion of it is not difficult. But you can Tstnfzh if your diet is rich in sodium, or if you are taking diuretics or use excessive laxatives.March exerciseYou can improve a lot of exercise, when doing exercises that increase heart rate, then return the heart to slow down the rate of pulses.The people who do not exercise have heart rate have about 80, and when they begin to practice some light exercises increase heart rate even up to 160, 170, and then after a short period of exercise, can be downloaded rate pulses to 60 and 65.The exercise will make you resist starting adrenaline overload.Like you graduated properly Tanatk, because you are using a starting adrenaline as part of their normal work.

Physical properties of blood

Physical properties of blood

Definition of blood Blood DefinitionBlood is a liquid texture, forms of connective tissue is happening inside the body within blood vessels (veins and arteries and capillaries) and consists of a liquid (plasma Plasma) swim blood cells Blood Corpuscles.Physical properties of bloodColor:Red color of blood and the presence of hemoglobin which gives blood on this color, red color varies in arteries than in veins is bright red in the arteries due to the presence of oxygen and dark red in the veins due to the presence of carbon dioxide.Temperature:It is fixed in the body, with some differences which from one member to another as needed for this member of the heat in order to do its main function. For example, the temperature of the liver equal (40-41) degree, either brain The degree temperature equal to less than 36 degrees Celsius.The overall rate of body temperature ranges between 36.8-37.8 degrees Celsius.Blood density:It relies on the presence of substances dissolved in the plasma, such as red blood cells, protein and value for men ranging from (1.057-1.067) g / cm 3.And women ranging between (1.051-1.061) g / cm 3.Blood viscosity:It is a friction force blood walls of the arteries and veins which mainly depend on the proteins found in plasma, especially Alvebrnugen and is important in maintaining blood pressure, a men's 4.7 and women 4.3.Osmosis:And this pressure results from the presence of crystals and minerals in the plasma, due importance to maintain the equivalent of salts and water inside and outside the cell (in the arteries and microvascular).For example, the presence of salts in abundance in the blood causes the withdrawal of water from the cells and this leads to dehydration, but the lack of salt water that causes entry into cells and this is known as water poisoning.
Pressure Alkoloido Asmosze (colloidal):And this pressure results from the presence of proteins in the plasma which is equal to 25 mm Hg and its importance due to:A) to maintain the presence of water within the vascular (blood volume).B) the exchange of nutrients between the blood and cells.A lack of protein in the blood leads to a lack of pressure Alkoloido and Asmosze which keeps water inside the cell and spoke edema Oedema and this pressure is less than the osmotic pressure normally.Density hydrogen concentration in the blood (PH):Usually this density tend to basal (ie blood basic solution) and is equal to this intensity of 7.4 and 7.35 in the arteries veins.The inside the cell is equal to (7-7.2) due to the presence of carbon dioxide.

Blood components

Blood components

 

1 - plasma plasmaIt is the liquid portion of blood, swim where blood cells and blood plasma ratio to total blood volume of 54%.Plasma properties:1. Color:Tends to yellow color and because of the presence of bilirubin Bilirubin.2. Figure:Plasma shapeless.3. Density:Plasma density is 1.027 g / cm 3, and it depends on the plasma proteins.4. The rest of the properties:(Viscosity / osmotic pressure, PH) as mentioned in the properties of blood.Blood plasma components:Blood plasma is composed of the following elements:• Water and accounts for 90% of the plasma volume.• solids and constitute 10% of the plasma volume, including:- 9% organic material.- 1% inorganic materials.1. Organic materials Organic materials:And this in turn is made up of:* Protein (6-8)% of the plasma volume (6-8 g / 100 cm 3 plasma).They are divided into:A - albumin Albumin and constitute 55% of the plasma proteins and this is equal to (8.3 to 1.5) g / 100 cm 3 plasma.B - globulin Globulin and accounting for 38% of the protein and this is equal to 3 g / 100 cm 3 plasma.C - Fabrinogen Fibrinogen and constitutes 7% of the protein and this equals (200-400) mg / plasma 100 cm 3.* Non-protein materials and are divided into:Food items, namely:- Sugars Glucose and increase of 80-120 mg / plasma 100 cm 3.- Lipid fats and increase of 600-800 mg / plasma 100 cm 3.Akhrajah materials such as:- Urea and urine increase of 11-53 mg / plasma 100 cm 3.- كرياتنين Creatinine increase of 0.8-1.2 mg / plasma 100 cm 3.- Uric acid Uric acid increase of 0.3-0.7 mg / plasma 100 cm 3.2. Inorganic materials Non organic materials:It includes:- Potassium and potassium 5.3 to 5.5 mg per Aikhuevilant / liter MEq / L- Sodium and sodium 135-153 mg per Aikhuevilant / liter.- Calcium and 8.8 -10.2 mg / plasma 100 cm 3.- Manganese and magnesium per 6.1 to 5.2 mg / plasma 100 cm 3.- Iron and iron rate of 100-150 mg / plasma 100 cm 3.- Chlorine and 38-110 mg per Aikhuevilant / liter.- Albaakrbunat.
2 - Blood cells Blood CellsAnd these cells are classified into:
Blood cells called because it does not contain recipes of cells where they contain a nucleus and nucleolus, Raibusomat and mitochondria so they are unable to divide and multiply.Numbering (5.4 to 5.6) million globule / mm 3 blood.
Blood cells in every sense of the word and number (4-11) A cell / mm 3 blood.• platelets Blood platelets.Circular objects do not carry recipes normal cell numbering (150-400) A plate / mm 3 blood.

Blood Circulation

Blood Circulation

Occur Arab world, Ibn al-Nafis (1210-1280) for the first time for micro-circulation and intended to promote blood circulation between the heart and lungs.After nearly four centuries came the British in 1678, "William Harvera" to talk in detail about great session and meant the circulation of blood between the heart and the rest of the body.The blood on a whirlwind tour of the human body, to reach every cell of it, carries the oxygen and food of any life, and carries waste, produced by and get rid of them through their work. On top of this waste carbon dioxide, which in turn get rid of it blood through a process of purification and cleaning take place in the lungs.And the blood passes through tours in the blood vessels of different capacities, size and composition.Called the vessels that carry blood from the heart to the other members of the body of the arteries, while called vessels that carry blood from the body to the heart veinsEveryone shall be two sessions: the bloody cycle or the circulatory system and other lymph.The circulatory system is divided blood or circulatory system into three parts all working at the same time:1 - Course pulmonary or Minor2 - plenary session or major session3 - Casement sessionMicro Valdorh or pulmonary circulation mobility blue blood to the lungs where it gets rid of carbon dioxide and provide themselves with oxygen. This is because blood red in color and loaded heart Booxgen.The plenary session of pure blood transfusion Red to the tissues.In textile filaments leaves blood carries oxygen and receives carbon dioxide and waste, to take her to and from the heart to the lungs through micro-session, which replaces the oxygen-carbon dioxide.The session Casement (PORTAL-CIRCULATION) are blood circulation to the liver, where moves blood received from the spleen, pancreas, stomach and intestines, and his liver converts glucose in the blood to glycogen and kept in reserve, come out of it in the case of need the body to him, as he turns Residue for cellular metabolism Albrootidi acid, urea, so The kidneys Bafrazha with urine.The center occupies the heart's main blood circulation. Reference is first and last a pump that receives blood and you pay to a permanent movement of vessels not only end with the end of life.Off blood from the heart, specifically the right part of it any of the right ventricle, through the pulmonary artery and pulmonary artery carrying blood saturated with carbon dioxide to the lungs Vivra to two major left and right.And branched These, in turn, to the branches reach each and every one of them to a certain lobe of the lung.And continues to branch like the branches of a tree even turned into a large network in harmony with lung Onsak (lung vesicles) Alveoles, where it is the process of purification and gas exchange.Gives blood how it surplus carbon dioxide and takes the oxygen it needs and returns through the veins, which are formed in a manner adverse to branch arteries, even to take final shape and make up four pulmonary veins carry blood saturated with oxygen to the heart, specifically the left atrium.This process is called the whole circulatory micro or small.What is different small for large session?Is essentially different from the major arteries that carry blood blue blood pregnant to carbon dioxide while carrying red blood veins purified oxygen dreamer, but in the session the major arteries that carry blood red while carrying blue blood veins.Off major blood during the session from the left ventricle through the aorta to the various parts of the body where branching artery bypass multiple names vary according to members that link to it and continue Baltafra even small branches up to every cell in the body.Up blood if, to the members of the body are all loaded with oxygen and return carrying carbon dioxide and other waste to accumulate through the veins that are formed in the same way that we have seen in the session Minor, and carry it to the heart through and Reden key are called veins Alogeoffan veines caves, one inferior vena cava vena cava inferior , gather the blood from the lower limbs to take it to the right atrium in the heart, while blood accumulates from the upper limbs by superior vena cava vena cava superior in turn also hurt in the right heart atrium.From there off to the right ventricle, where pumped through the pulmonary artery to the lungs to begin Minor new session.


It should be noted that the blood is subject during its grand to the purification process is in the kidneys, where blood is free from some of the material excreted with urine through a complex process of absorption and excretion.As is also subject to the process of purifying the other is in the liver through the cycle generic form part of the session the major, and are made through the portal vein (portal vein), which is transmitted through which blood coming from the intestines and abdominal organs other and which also carries food gained from foods that are undergoing liver where it is receive. فالكبد as we know is the chemical lab in the body.The sessions Aldmoatin Mvsoltan completely from each other only in some diseases that remain a hole or more between the wall of the atrium or the right ventricle Tsalhma Bashbahiama Aloasrin.Some of these diseases lead to congenital cyanosis, and sometimes remain the link between the pulmonary artery and the artery Alarbehr, called the ductus arteriosus, impassable which also leads to mixing of blood blue blood red.

Acute Lymphocytic Leukemia

Acute Lymphocytic Leukemia

Leukemia is cancer of the white blood cells. White blood cells help your body fight infection. Your blood cells form in your bone marrow. In leukemia, however, the bone marrow produces abnormal white blood cells. These cells crowd out the healthy blood cells, making it hard for blood to do its work. In acute lymphocytic leukemia (ALL), there are too many of specific types of white blood cells called lymphocytes or lymphoblasts. ALL is the most common type of cancer in children.
Possible risk factors for ALL include being male, being white, previous chemotherapy treatment, exposure to radiation, and for adults, being older than 70.
Symptoms of ALL include:

• Weakness or feeling tired
• Fever
• Easy bruising or bleeding
• Bleeding under the skin
• Shortness of breath
• Weight loss or loss of appetite
• Pain in the bones or stomach
• Pain or a feeling of fullness below the ribs
• Painless lumps in the neck, underarm, stomach, or groin

Tests that examine the blood and bone marrow diagnose ALL. Treatments include chemotherapy, radiation therapy, stem cell transplants, and targeted immune therapy. Once the leukemia is in remission, you need additional treatment to make sure that it does not come back.

NIH: National Cancer Institute

Book on Biological Laboratory penalty 17

Book on Biological Laboratory penalty 17

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Policy & Procedure Manual
Policy # MI\TECH\48\v01  Page 3 of 3
Technical Manual   

Precautions

1.  The TSI tube should be read within 18-24 hr.  If read earlier, a false +/+ reaction may occur;
if after 24 hr, a fals e -/-reaction may occur.

2.   An H2S organism may produce so much black precipitate that the acidity in the butt is
completely masked.  If H
2
S is produced, an acid cond ition exists in the butt.

3.  There is no inhibitor in this  medium, therefore any organism  may grow.  Be certain that the
organism tested is a  catalase positive, gram  negative bacillus.

4.  S. typhi  usually produces a ring of H
2
S near the surface of the butt.  Occasionally the amount
of H
2
S produced is so small that it  will not be detect ed in TSI, but will show up in SIM
media.

5.  Some organisms produce such an abundance of gas that th e medium may be completely
displaced by gas, resulting in the medium being blown up into the cap of the tube.  Use
caution to avoid contamination.

6.  Do not tighten the cap of a TSI tube.  A free exchange of air is necessary to enhance the
alkaline reaction of the slant.

Quality Control

Test the media each  time it is prepar ed using the following organisms:

E. coli:    (ATCC 25922) :  +/+
  P. mirabilis:  (ATCC 12453) :  -/+/H2
S
  P. aeruginosa: (ATCC 27853) :  -/-

References

1. MacFaddin JF, Bioche mical Tests for Identifi cation of Medical Bacteria, 2nd ed., Williams
and Wilkins, Baltimore MD, 1980, p183-194.

PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT 

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Policy & Procedure Manual
Policy #MI\TECH\49\v01  Page 1 of  2
Section: Technical Manual  Subject Title: Tube Coagulase Test
Issued by:  LABORATORY MANAGER   Original Date: July 31, 2000
Approved by: Laboratory Director

Revision Date: February 15, 2002

TUBE COAGULASE TEST
Principle

This test is used to speciate staphylococci by determ ining the ability of an isol ate to clot plasma by
producing the enzyme coagulase.

Reagents

Rabbit plasma 

  1.  Reconstitute one vial at  a time with sterile distilled water (volume determined by
vial size).

  2.  Store refrigerated before  and after reconstitution.   Use within 72 hours of
reconstitution.

Other Materials

Sterile glass tubes (tube method)
Culture loop or wooden applicator stick

Procedure

  1.  Add 0.5 mL of  plasma to a sterile glass tube.

  2.  Emulsify a large loop ful of a pure colony of  Staphylococcus into the plasma.

  3.  Incubate at 35
o
C for 4 hr, observing every 30  minutes for clot formation.

4.   If there is no visible clot at the end of 4  hours, leave at room temperature overnight
and observe for cl ot formation.

Interpretation

  Positive: Clot formation

  Negative: No clot formation

PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT 

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Policy # MI\TECH\49\v01  Page 2 of 2
Technical Manual   

Precautions

1)  When observing the tube, do not shake or agitate the tube.

Quality Control

Each time a coagulase test is performed, known  positive and negative cultures must be tested.

 Positive: S. aureus  (ATCC 25923)
 Negative: S. epidermidis  (ATCC 12228)

References

1.  MacFaddin, J.F., Biochemical Tests for Identification of Medical Bacteria, 2nd ed.,
Williams and Wilkins, Baltimore MD, 1980, pgs. 64-77.

PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT 

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TML/MSH Microbiology Department
Policy & Procedure Manual
Policy #MI\TECH\50\v01  Page1 of  2 
Section: Technical Manual  Subject Title: Urea Slant
Issued by:  LABORATORY MANAGER   Original Date: July 31, 2000
Approved by: Laboratory Director

Revision Date: February 15, 2002

UREA SLANT

Principle

To determine the ability of an organism to split urea by the action of the enzyme urease forming
two molecules of ammonia with resulting alkalinity.

Materials

Urea Slant
Bacteriology loop

Procedure

1.   From one isolated colony, heavily inoculate the urea slant.
2.   Incubate O
2, 35
0
C.
3.   Read at 3 hours and ag ain at 18-24 hours.

Interpretation

Positive: Intense pink-red  colour.
    Rapidly positive:  1 to 6 hours (Proteus spp.)
  Delayed positive: ≥ 18 hours

Negative: No colour change

Precautions

Urea test media rely on the demonstration of alkalinity, thus are not specific for urease.  The
utilization of peptones or other prot eins may cause an increase in pH.

Quality Control

Controls should be set up weekly.

P. mirabilis  (ATCC 12453):    Positive - 4 hours
  K. pneumoniae (ATCC 13883):  Weak positive - 18 hours
  E. coli  (ATCC 25922):    Negative

PROCEDURE MANUAL
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Page 131
         
TML/MSH Microbiology Department
Policy & Procedure Manual
Policy # MI\TECH\50\v01  Page 2 of 2
Technical Manual   

References

1.  MacFaddin JF, Biochemical Tests for Identification of Medical Bacteria, 2
nd
 ed.,
Williams and Wilkins, Baltimore MD, 1980, p298-308.


PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT 

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TML/MSH Microbiology Department
Policy & Procedure Manual
Policy #MI\TECH\51\v01  Page 1 of  2
Section: Technical Manual  Subject Title: X And V Disks for
  Identification of Haemophilus
Issued by:  LABORATORY MANAGER   Original Date: July 31, 2000
Approved by: Laboratory Director

Revision Date: February 15, 2002

X AND V DISKS FOR IDENTIFICATION OF HAEMOPHILUS

Principle

Haemophilus spp. have different requir ements for the growth factor s X (hemin) and V (NAD).
These requirements are determined based on th e presence or absence of growth around disks
impregnated with V,X and XV factors.

Reagents

1.  Bacto Differentiation Disks
    BV  NAD and 125 units/ml bacitracin
    BX  hemin and 125 units/ml bacitracin
    BVX NAD, hemin  125 units/ml bacitracin
  Store refrigerated

2.  Mueller Hinton Agar (MHA)

Other Materials

Forceps
Swabs
Inoculating loop

Procedure

1.  Pick one colony from CHOC, taking care not to carry over  any agar from the medium.
2.  Inoculate MHA and streak  over the entire surface of th e plate using a sterile swab.
3.  Place X, V and XV disks on the surface of the agar in the form of a triangle with at least 30-35 mm between disks.
4. Incubate CO
2
, 35
o
C X 18-24 hr.
5.  Examine the pattern of growth around and/or between the disks.

Interpretation

Growth around the V and XV or the X and XV indicates a requirement for the single growth factor
V or X respectively.  Growth around only the XV disk indicates a requiremen t for both factors.

PROCEDURE MANUAL
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Policy & Procedure Manual
Policy # MI\TECH\51\v01  Page 2 of  2
Technical Manual   

Precautions

1.  Avoid carry-over of growth factors.

Quality Control

Known positive and negative controls must be set up in parallel with the test.

H. influenzae (ATCC 35056):    Growth  around the XV disk only
H. parainfluenzae (ATCC 7901):   Growth around V and XV disks

References

1. Difco Package in sert, June 1984.


PROCEDURE MANUAL
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TML/MSH Microbiology Department
Policy & Procedure Manual
Policy #MI\TECH\52\v01  Page 1 of  1
Section: Technical Manual  Subject Title: Xylose Fermentation
Issued by:  LABORATORY MANAGER   Original Date: July 31, 2000
Approved by: Laboratory Director

Revision Date: February 15, 2002

XYLOSE FERMENTATION
Principle

A rapid chromogenic test for the identification of  E. gallinarum.

Reagents

Prolab d-xylose tablets.
Sterile staline.

Other Materials

Sterile tubes (13 x 100 mm)

Procedures

1.   Suspend the growth from BA in 0.25 mL salin e to achieve the turbidity >#2 McFarland
standard.
2.   Add 1 tablet to the tube.
3.   Incubate at 35 - 37
0
C x 2 hours.
4.   Examine the tube for development of a yellow colour.

Interpretation

Positive:  Yellow / yellow orange colour
Negative: Red

Quality Control

The following organisms are tested weekly:

 Positive: E. gallinarum (ATCC 35038)
 Negative: E. faecalis (ATCC 29212)

References

1.  J. Clin. Microbiol. 12, 620-623, 1980.

Book on Biological Laboratory penalty 16

Book on Biological Laboratory penalty 16

Page 122

TML/MSH Microbiology Department
Policy & Procedure Manual
Policy #MI\TECH\46\v01  Page 1 of  2
Section: Technical Manual  Subject Title: Thermonuclease Test
Issued by:  LABORATORY MANAGER   Original Date: July 31, 2000
Approved by: Laboratory Director

Revision Date: February 15, 2002

THERMONUCLEASE TEST
Principle

Staphylococcus aureus  contains a heat-stable thermonu clease and coagulase negative
staphylococcus does not.  This is a rapid test to differentiate between the two organisms.

Materials

Toluidine blue-O  DNA plate (Q-Lab)
13x100 mm tube with white cap
pasteur pipettes

Procedure

1.  Dispense 2 - 3 mL of blood  broth from BacT/Alert bottle  showing gram po sitive cocci in
clusters in the direct Gram stain into a sterile capped 13x100 mm tube.
2.  Place tube in heating block, 100
0
C for 15 minutes.
3.  Let cool to room temperature.
4.  Centrifuge at approximat ely 2500 rpm for 3 minutes.
5.  Inoculate a pre-warmed (35
o
C for 1 hour) toluidine blue-O  DNA plate by filling wells (cut
well with the end of a pasteur pipette) with 2 drops of the supernatant.
6.  Incubate the plate at 35
o
C in the upright positio n (agar side down).
7.  Inspect the plate at, 1 hour, 2 hours and 4 hours and again  after overnight incubation if
negative at 4 hours.
8.  Always run negative and positive cont rol wells with each  plate each day.

Interpretation

Positive:  Pink zone of clearing at the edge of  the well with a darker  blue ring at the outer
periphery of the zone;  indi cates thermonuclease activity

Negative:  No zone or a small clear zone around the well

PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT 

Page 123

TML/MSH Microbiology Department
Policy & Procedure Manual
Policy # MI\TECH\46\v01  Page 2 of 2
Technical Manual   

Quality Control

1.  Inoculate 5 day negative patient BacT/Alert bottles with 0.5 mL of  a slightly turbid
suspension of (a)  S. aureus  (ATCC 25923) and (b)  S. epidermidis  (ATCC 12228) in
trypticase soy broth.

2. Incubate the bo ttles overnight at 36
o
C on the shaker.

3.  Remove 3 - 6 mL of the broth-blood from the bottles and process in the same manner as the
patient specimens (steps 1 to 4). Always QC new controls before use with patient specimen.

4.         Supernatants may be kept refriger ated for up to 1 month for use as controls.

Reference

1.  Rafner, H.B., & Stretton C.W.  1985.  Thermonuclease test fo r same day identification of  S.
aureus  in blood cultures.  J.  Clin. Microbiol. 21:995-996.


PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT 

Page 124

TML/MSH Microbiology Department
Policy & Procedure Manual
Policy # MI\TECH\47\v01  Page 1 of  1
Section: Technical Manual  Subject Title: Tributyrin Test
Issued by:  LABORATORY MANAGER   Original Date: July 31, 2000
Approved by: Laboratory Director

Revision Date: February 15, 2002

TRIBUTYRIN TEST
Principle

A rapid chromogenic test for the identification of M. catarrhalis.

Reagents

Prolab Tributyrin (TRIB) tablets.
  Store at room temperature.
Sterile saline.

Other Materials

Sterile tubes (13 x 100 mm)

Procedures

1.  Suspend the growth from C HOC in 0.25 mL (6 drops) saline to achieve the  turbidity >#2
McFarland standard.
2.  Add 1 tablet to the tube.
3.  Incubate at 35
0
C x 4 hours.
4.  Examine the tube for development of a yellow colour.

Interpretation

Positive: Yellow/yellow orange colour

Negative: Red

Quality Control

Test the following organism weekly:

Positive:  M. catarrhalis  (ATCC 8176) 
 Negative: N. gonorrhoeae (ATCC 43069)
 
References

1.  Prolab package insert, February 1985.

PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT

Page 125

TML/MSH Microbiology Department
Policy & Procedure Manual
Policy #MI\TECH\48\v01  Page 1 of  3
Section: Technical Manual  Subject Title: TSI (Triple Sugar Iron)
Issued by:  LABORATORY MANAGER   Original Date: July 31, 2000
Approved by: Laboratory Director

Revision Date: February 15, 2002

TSI (TRIPLE SUGAR IRON)
Principle

To determine the ability of an organism to attack  a specific carbohydrate  incorporated in a basal
growth medium, with or without  the production of gas, along with  the determination of possible
hydrogen sulfide (H
2
S) production.  This test is used,  in conjunction with others, for the
identification of enteric pathogens.

Materials

TSI Slant
Inoculating wire or sterile glass pasteur pipette.

Procedure

1.  Using the an inoculating wire, dip into the previously inoculated TSB.
2.  Stab the butt of the TSI to within 1/4 inch from bo ttom, draw out and fishtail over slant.  Do
not tighten cap.
3. Incubate O
2, 35
o
C X 18-24 hours.

Interpretation

Carbohydrate utilization:
  1.  Fermentation of glucose only
    (a)  slant:  red colour (alkaline reaction)
    (b)   butt:   yellow colour (acid reaction)

  2.  Fermentation of glucose and sucrose and/or lactose
    (a)  slant:  yellow colour (acid reaction)
    (b)  butt:   yellow colour (acid reaction)

  3.  Neither glucose nor lactose nor sucrose fermented
    (a)  slant:  red colour (alkaline reaction)
  (b) butt: (i)  aerobic organism
        (a)  No growth
        (b)  No colour change
 (ii) facultative organism       red colour (alkaline reaction)

PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT

Page 126

TML/MSH Microbiology Department
Policy & Procedure Manual
Policy # MI\TECH\48\v01  Page 2 of 3
Technical Manual  

Gas production:

 1. Aerogenic:
  (a) Gas production:  CO2 and H
2

    (b)  Evident by one of the following:
    (i) a single gas bubble
    (ii) bubbles in the medium
             (iii)  splitting of medium
    (iv) complete displacement of the medium from bottom of the tube
leaving a clear area
        (v)  slight indentation of medium from the side of the tube

 2. Anaerogenic:
   No gas production
 
H2
S production:

      The presence of a black precipita te (ferrous sulfid e) is evident by:
(i)   A black colour spread throughout the entire butt masking the
            acidity;  may even  be a slight evidence  on the slant
     (ii) A black ring near the top of the butt area
  (iii)      A black precipitate scattered throughout  the butt  but not entirely
                        masking the acidity present
Summary:

  The ways of recording the TSI reactions are list ed below.  Reme mber that the slant is first,
followed by the butt reaction.

  acid/acid    +/+
  acid/acid/gas    +/+ with gas
  acid/acid/gas/H2S   +/+ with H 2
S
  alkaline/acid    -/+
  alkaline/acid/gas   -/+ with gas
  alkaline/acid/gas/H2S      -/+ with gas and H2
S
  alkaline/acid/H2S        -/+ with H 2
S
  alkaline/alkaline   -/-


PROCEDURE MANUAL
TORONTO MEDICAL LABORATORIES / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT