1

Chapter 18Sputum AnalysisProfessor A. S.

Alhomida

Chapter 18Sputum AnalysisProfessor A. S.

Alhomida

DisclaimerDisclaimer• The texts, tables and images contained in this course The texts, tables and images contained in this course

presentation (BCH 376) are not my own, they can be found on: presentation (BCH 376) are not my own, they can be found on: • References suppliedReferences supplied• Atlases orAtlases or• The webThe web

King Saud University

College of Science

Department of Biochemistry

2

1.1. It is a secretion that is produced in the lungs It is a secretion that is produced in the lungs and the bronchi (tubes that carry the air to and the bronchi (tubes that carry the air to the lung), also is known as phlegmthe lung), also is known as phlegm

2.2. This mucus-like secretion may become This mucus-like secretion may become infected, bloodstained, or contain abnormal infected, bloodstained, or contain abnormal cells that may lead to a diagnosiscells that may lead to a diagnosis

Sputum DefinitionSputum Definition

3

Sputum Definition, Cont’dSputum Definition, Cont’d

3.3. Tracheobronchial sections are an Tracheobronchial sections are an inconstant mixture of plasma, water, inconstant mixture of plasma, water, electrolytes and mucinelectrolytes and mucin

4.4. As these mixture pas through the lower and As these mixture pas through the lower and upper respiratory tract, they become upper respiratory tract, they become contaminated with cellular exfoliations, contaminated with cellular exfoliations, nasal and salivary gland secretions and nasal and salivary gland secretions and normal bacterial flora of the oral cavitynormal bacterial flora of the oral cavity

4

Normal LungNormal Lung

• Trachea is split up into Trachea is split up into two bronchus that leads two bronchus that leads into both our lungsinto both our lungs

• The bronchi are further The bronchi are further distributed into bronchi distributed into bronchi and bronchiole and bronchiole

• The air travels along The air travels along these tubes and finally these tubes and finally ends up in the air sacs ends up in the air sacs (alveoli) (alveoli)

• Alveoli are surrounded by Alveoli are surrounded by a network of capillariesa network of capillaries

5

Normal Lung, Cont’dNormal Lung, Cont’d

• The major features of the The major features of the lungs include the bronchi, lungs include the bronchi, the bronchioles and the the bronchioles and the alveolialveoli

• Each alveolus is Each alveolus is surrounded by a dense bed surrounded by a dense bed of capillariesof capillaries

• The alveoli are the sites of The alveoli are the sites of the actual gas exchange the actual gas exchange and are regarded as the and are regarded as the primary functional units of primary functional units of the lungs the lungs

6

Normal Lung, Cont’dNormal Lung, Cont’d

• OO22 entering an alveolus entering an alveolus dissolves in the film of water dissolves in the film of water on its wall and moves by on its wall and moves by diffusion across the cells into diffusion across the cells into the bloodthe blood

• OO22 is in higher concentration is in higher concentration inside the alveolus than in the inside the alveolus than in the blood blood

• OO22 moves from a higher moves from a higher concentration to a lower concentration to a lower concentration by the process concentration by the process of diffusion of diffusion

• Concentration of COConcentration of CO22 is higher is higher in the blood than in the alveoli in the blood than in the alveoli

• COCO22 also moves from a high also moves from a high concentration to a lower concentration to a lower concentration by the process concentration by the process of diffusion of diffusion

7

1. Drinking a lot of water and other fluids the night before the test may help to get the sample

2. To be asked to cough deeply and spit any sputum in a sterile cup

3. The sputum is then taken to the laboratory4. There, it is placed in a special substance

(medium) under conditions that allow the organisms to grow

Sputum CollectionSputum Collection

8

Sputum CollectionSputum Collection

1. Drinking a lot of water and other fluids the night before the test may help to get the sample

2. To be asked to cough deeply and spit any sputum in a sterile cup

3. The sputum is then taken to the laboratory

4. There, it is placed in a special substance (medium) under conditions that allow the organisms to grow

9

Sputum AnalysisSputum Analysis

• This slide shows typical This slide shows typical buccal squamous epithelial buccal squamous epithelial cells which are much larger cells which are much larger than polymorphonuclear than polymorphonuclear leukocytes (PMN) and take leukocytes (PMN) and take up most of the field in a up most of the field in a high power viewhigh power view

• This cell is covered with This cell is covered with chains of Gram positive chains of Gram positive cocci typical of normal oral cocci typical of normal oral flora such as flora such as peptostreptococcipeptostreptococci. (oil . (oil immersion, 1000x)immersion, 1000x)

10

Physical Properties of Physical Properties of SputumSputum

1.1. AppearanceAppearance• It may be described as liquid (serous), It may be described as liquid (serous),

mucoil, purulent, bloody or combination of mucoil, purulent, bloody or combination of thesestheses

2.2. ColorColor• Its color is determined by the material Its color is determined by the material

contained, and often color can indicate the contained, and often color can indicate the pathological processpathological process

• Yellow color indicates pus and epithelial Yellow color indicates pus and epithelial cellscells are present are present

11

3.3. OdorOdor• Usually no odor is present in normal and Usually no odor is present in normal and

pathological sputum, butpathological sputum, but if bacterial if bacterial decomposition has been taken place decomposition has been taken place within the body or after expectoration, a within the body or after expectoration, a variety of odor will be presentvariety of odor will be present

Physical Properties of Physical Properties of Sputum, Cont’dSputum, Cont’d

12

Miscellaneous Findings in Miscellaneous Findings in SputumSputum

1.1. Cheesy MassesCheesy Masses • These are fragments of necrotic These are fragments of necrotic

pulmonary tissue seen in such disease pulmonary tissue seen in such disease as pulmonary gangrene or tuberculosisas pulmonary gangrene or tuberculosis

2.2. Bronchial CastsBronchial Casts• These are branching tree like casts of These are branching tree like casts of

bronchi whose size varies with that of bronchi whose size varies with that of bronchi in which they are formedbronchi in which they are formed

• They are composed of fibrin and are They are composed of fibrin and are white or gray colorwhite or gray color

13

3.3. Broncholiths (Lung Stones)Broncholiths (Lung Stones)• They are formed by calcification of They are formed by calcification of

necrotic or infected tissuesnecrotic or infected tissues• Chronic tuberculosis is the most common Chronic tuberculosis is the most common

casue for their formationcasue for their formation

Miscellaneous Findings in Miscellaneous Findings in Sputum, Cont’dSputum, Cont’d

14

4.4. Dietrich's PlugsDietrich's Plugs• They are frequently observed in putrid They are frequently observed in putrid

bronchitis and bronchiectosisbronchitis and bronchiectosis• They are composed of cellular debris, fatty They are composed of cellular debris, fatty

acids, crystals, fat globules and bacteriaacids, crystals, fat globules and bacteria

Miscellaneous Findings in Miscellaneous Findings in Sputum, Cont’dSputum, Cont’d

15

5.5. Foreign BodiesForeign Bodies• In children, they can be any small object a In children, they can be any small object a

child may place it into his mouthchild may place it into his mouth• In adults, they are either food particles or In adults, they are either food particles or

gastric contents aspirated during convulsion, gastric contents aspirated during convulsion, during intoxication or operative anesthesiaduring intoxication or operative anesthesia

6.6. ParasitesParasites• They are extremely rare They are extremely rare

Miscellaneous Findings in Miscellaneous Findings in Sputum, Cont’dSputum, Cont’d

16

Sputum Chemical Sputum Chemical CompositionComposition

ComponentComponent % of Total % of Total WeightWeight

WaterWater 9595

SolidSolid 55

% of Total Solids% of Total SolidsCarbohydratesCarbohydrates VariableVariable

ProteinsProteins VariableVariable

LipidsLipids VariableVariable

DNADNA VariableVariable

Enzymes, Enzymes, -antitrypsin, -antitrypsin, LDH, lysozyme, LDH, lysozyme, lactoferrinlactoferrin

VariableVariable

17

1.1. Pneumonia occurs when the lungs become Pneumonia occurs when the lungs become inflamed and infectedinflamed and infected. . Because the lungs are Because the lungs are infected, pneumonia makes it difficult to breathinfected, pneumonia makes it difficult to breath. . It often follows a common cold or the fluIt often follows a common cold or the flu

2. Although people of all ages can get pneumonia, it is especially dangerous for people with chronic illnesses, very young children, and older adults

Definition of PneumoniaDefinition of Pneumonia

18

3. Pneumonia can be caused by either bacteria, fungi, amoebae, parasites or a virus.

4. One or both lungs can be affected

Definition of Pneumonia, Definition of Pneumonia, Cont’dCont’d

19

Pneumonia, Cont’dPneumonia, Cont’d

• The alveoli filled with The alveoli filled with mucus such as submucosa mucus such as submucosa widened by smooth muscle widened by smooth muscle hypertrophy, edema, and hypertrophy, edema, and inflammation inflammation ((mainly mainly eosinophilseosinophils) )

• These are changes of These are changes of bronchial asthmabronchial asthma

• The peripheral eosinophil The peripheral eosinophil count or the sputum count or the sputum eosinophils can be eosinophils can be increased during an increased during an asthmatic attackasthmatic attack..

20

Pneumonia, Cont’dPneumonia, Cont’d

21

Pneumonia, Cont’dPneumonia, Cont’d

22

Symptoms of PneumoniaSymptoms of Pneumonia

• Pneumonia symptoms vary depending upon Pneumonia symptoms vary depending upon the type of pneumonia. Symptoms may the type of pneumonia. Symptoms may include:include:• Fever Fever • Chest pain and/or muscle pain Chest pain and/or muscle pain • Cough that may produce white, yellow, or Cough that may produce white, yellow, or

green mucus (phlegm) green mucus (phlegm) • Lack of energy Lack of energy

23

• White nail syndrome may also be called leukonychia

• Leukonychia can occur with arsenic poisoning, heart disease, renal failure, pneumonia, or

• Hypoalbuminemia

Symptoms of PneumoniaSymptoms of Pneumonia

24

Sputum Analysis: Sputum Analysis: PneumoniaPneumonia

• Moraxella catarrhalisMoraxella catarrhalis, a , a large number of Gram large number of Gram negative (red) cocci are negative (red) cocci are seen and many appear to seen and many appear to be attaching to or be attaching to or residing within the PMNsresiding within the PMNs

• Some physicians confuse Some physicians confuse these organisms with the these organisms with the Gram negative Gram negative coccobacillarycoccobacillary

25

Sputum Analysis: Pneumonia, Sputum Analysis: Pneumonia, Cont’dCont’d

• HemophilusHemophilus influenzae influenzae pneumonia pneumonia demonstrating the demonstrating the typical Gram negative typical Gram negative coccobacillary formscoccobacillary forms

• Because of the red Because of the red background produced by background produced by the Gram stain method, the Gram stain method, these organisms can be these organisms can be difficult to see (oil difficult to see (oil immersion, 1000ximmersion, 1000x))

26

Sputum Analysis: Pneumonia, Sputum Analysis: Pneumonia, Cont’dCont’d

• HemophilusHemophilus influenzae influenzae pneumoniapneumonia

27

Sputum Analysis: Pneumonia, Sputum Analysis: Pneumonia, Cont’dCont’d

• Klebsiella Klebsiella pneumoniaepneumoniae pneumonia pneumonia demonstrating Gram demonstrating Gram negative bacillary negative bacillary organisms. (oil organisms. (oil immersion, 1000x)immersion, 1000x)

28

Sputum Analysis: Pneumonia, Sputum Analysis: Pneumonia, Cont’dCont’d

• Gram stain of the Gram stain of the sputum from a patient sputum from a patient with with StaphylococcusStaphylococcus aureus pneumonia aureus pneumonia demonstrating clusters demonstrating clusters of Gram positive cocci of Gram positive cocci some of which are some of which are associated with the associated with the PMNs (oil immersion, PMNs (oil immersion, 1000x)1000x)

29

Sputum Analysis: Pneumonia, Sputum Analysis: Pneumonia, Cont’dCont’d

• GramGram--positive, positive, spherical bacteria, spherical bacteria, Streptococcus Streptococcus pneumoniaepneumoniae, is the , is the cause of many cause of many human diseases, human diseases, including pneumoniaincluding pneumonia

30

Treatments of PneumoniaTreatments of Pneumonia

1.1. Treatments depend upon the type of Treatments depend upon the type of pneumonia and the seriousness of the pneumonia and the seriousness of the symptomssymptoms

2.2. Bacterial pneumoniaBacterial pneumonia is usually treated with is usually treated with antibiotics. antibiotics. Viral pneumoniaViral pneumonia is not treated is not treated with antibiotics but with rest, fluids, and with antibiotics but with rest, fluids, and time. Recovery for viral pneumonia is time. Recovery for viral pneumonia is usually longer than bacterial pneumoniausually longer than bacterial pneumonia

31

Treatments of Pneumonia, Treatments of Pneumonia, Cont’dCont’d

3.3. Doctor may also recommend taking over-Doctor may also recommend taking over-the-counter medicines to reduce fever and the-counter medicines to reduce fever and treat aches and pains. Because coughing is treat aches and pains. Because coughing is important to help clear your lungs, it is important to help clear your lungs, it is important to avoid cough suppressantsimportant to avoid cough suppressants

32

Definition of AsthmaDefinition of Asthma1.1. It is a chronic lung condition that is It is a chronic lung condition that is

characterized by a difficulty in breathing. A characterized by a difficulty in breathing. A person with asthma will have extremely person with asthma will have extremely sensitive airways that react by becoming sensitive airways that react by becoming narrow when agitatednarrow when agitated

2.2. Often, asthma is confused with allergies, such Often, asthma is confused with allergies, such as the wheezing and coughing that comes when as the wheezing and coughing that comes when a person is allergic to pollen and dust in the aira person is allergic to pollen and dust in the air

3.3. Too much exposure to allergens can lead to the Too much exposure to allergens can lead to the development of asthmadevelopment of asthma

33

Definition of Asthma, Definition of Asthma, Cont’dCont’d

3.3. The allergens cause the immune system to The allergens cause the immune system to kick into high gear in the same way it would if kick into high gear in the same way it would if fighting off a virus or parasitefighting off a virus or parasite

4.4. For most people, this reaction induces For most people, this reaction induces coughingcoughing

5.5. But if the airways become inflamed, and if it But if the airways become inflamed, and if it happens with frequency, the body reacts with happens with frequency, the body reacts with wheezing and general difficulty in breathingwheezing and general difficulty in breathing

34

Normal and Asthmatic Normal and Asthmatic BronchioleBronchiole

• When an asthma attack When an asthma attack occurs, mucus occurs, mucus production is increasedproduction is increased

• Muscles of the bronchial Muscles of the bronchial tree become tight, and tree become tight, and the lining of the air the lining of the air passages swells, passages swells, reducing airflow and reducing airflow and producing the producing the characteristic wheezing characteristic wheezing soundsound

35

Normal and Asthmatic Normal and Asthmatic BronchioleBronchiole

• During an asthma attack During an asthma attack smooth muscles located smooth muscles located in the bronchioles of the in the bronchioles of the lung constrict and lung constrict and decrease the flow of air decrease the flow of air in the airwaysin the airways

• The amount of air flow The amount of air flow can further be can further be decreased by decreased by inflammation or excess inflammation or excess mucus secretionmucus secretion

36

Symptoms of AsthmaSymptoms of Asthma

1.1. Shortness of breathShortness of breath2.2. Wheezing (a whistling noise in the chest) Wheezing (a whistling noise in the chest) 3.3. Cough and chest tightnessCough and chest tightness

• Not everybody will have all these symptomsNot everybody will have all these symptoms• Young children often present only with a cough Young children often present only with a cough

with no other symptoms hence often the asthma with no other symptoms hence often the asthma diagnosis is overlooked or misdiagnoseddiagnosis is overlooked or misdiagnosed

• A history of asthma, eczema or hay fever in the A history of asthma, eczema or hay fever in the family may mean that your chances of family may mean that your chances of developing asthma are slightly higher than developing asthma are slightly higher than those withoutthose without

37

Process of AsthmaProcess of Asthma

38

Mechanism of Asthma Mechanism of Asthma AttackAttack

39

Mechanism of Asthma Mechanism of Asthma Attack, Cont’dAttack, Cont’d

40

Asthma, Cont’dAsthma, Cont’d

41

Asthma, Cont’dAsthma, Cont’d

42

Provokes AsthmaProvokes Asthma• It attacks are provoked by two factors: It attacks are provoked by two factors:

• Triggers, which result in Triggers, which result in bronchoconstriction (tightening of the bronchoconstriction (tightening of the airways), or airways), or

• Causes, which result in inflammation of Causes, which result in inflammation of the airwaysthe airways

43

Analysis of Sputum: Analysis of Sputum: AsthmaAsthma

• Note thick basal Note thick basal membrane under the membrane under the epithelial layer, epithelial layer, which is covered which is covered with much mucus in with much mucus in fatal case of status fatal case of status asthmaticusasthmaticus

44

Analysis of Sputum: Analysis of Sputum: Asthma, Cont’dAsthma, Cont’d

• Small airways show Small airways show considerable hyperplasia considerable hyperplasia and hypertrophy of smooth and hypertrophy of smooth muscle, goblet cells and muscle, goblet cells and thickening of the mucosathickening of the mucosa

• The airway wall is covered The airway wall is covered with unusually large with unusually large amounts of mucus, and amounts of mucus, and projects into the lumen projects into the lumen like villilike villi

• With minimal additional With minimal additional smooth muscle shortening smooth muscle shortening or edema the airway is or edema the airway is fully closedfully closed

45

Analysis of Sputum: Analysis of Sputum: Asthma, Con’tdAsthma, Con’td

• This very large spiral was This very large spiral was found in lavage fluid from found in lavage fluid from an asthmatic patient an asthmatic patient being evaluated for a lung being evaluated for a lung mass. mass.

• The Papanicolaou stain The Papanicolaou stain gives a reddish core with gives a reddish core with a fuzzy margin to which a fuzzy margin to which many cells are attached many cells are attached secondarilysecondarily

• The arrow points out a The arrow points out a macrophage for macrophage for comparison of size. The comparison of size. The spiral is about 1 mm longspiral is about 1 mm long

46

Causes of AsthmaCauses of Asthma

• They are also known as inducers, cause They are also known as inducers, cause airway inflammation and airway airway inflammation and airway hyperresponsiveness, both causes of asthmahyperresponsiveness, both causes of asthma

• The most common causes are allergens and The most common causes are allergens and viral respiratory illnessviral respiratory illness

47

Asthma TriggersAsthma Triggers

• Many of the same substances that trigger allergies can also trigger asthma

• Common allergens include pollen, dust mites, mold and pet dander

• Other asthma triggers include irritants like smoke, pollution, fumes, cleaning chemicals, and sprays

• Asthma symptoms can be substantially reduced by avoiding exposure to known allergens and respiratory irritants

48

Treatments of AsthmaTreatments of Asthma• There are different types of treatment for asthma There are different types of treatment for asthma

suffererssufferers• The type of treatment depends on the severity of the The type of treatment depends on the severity of the

asthma and its attacksasthma and its attacks• Asthma may not be able to be cured, but it can be Asthma may not be able to be cured, but it can be

controlledcontrolled• By controlling the asthma, people who suffer from the By controlling the asthma, people who suffer from the

disease are able to live relatively normal, active livesdisease are able to live relatively normal, active lives

49

1.1. AntihistaminesAntihistamines

• For the person who suffers allergies along with For the person who suffers allergies along with asthma, the doctor might prescribe antihistamines asthma, the doctor might prescribe antihistamines to help control the reaction to allergensto help control the reaction to allergens

• Antihistamines are not without their downsideAntihistamines are not without their downside• They can cause drowsiness, limiting a person’s They can cause drowsiness, limiting a person’s

ability to work at peak level, and also they can also ability to work at peak level, and also they can also cause dry mouth, urinary problems, and cause dry mouth, urinary problems, and constipationconstipation

Treatments of Asthma, Treatments of Asthma, Cont’dCont’d

50

2.2. BronchodilatorsBronchodilators

• It helps to relax the muscle of the bronchial tubes, It helps to relax the muscle of the bronchial tubes, making breathing easier. There are several different making breathing easier. There are several different bronchodilators: bronchodilators: -agonist, theophylline, and -agonist, theophylline, and anticholinergicsanticholinergics

• -agonist bronchodilators can be short-term or long--agonist bronchodilators can be short-term or long-term treatmentsterm treatments• The short-term treatments are used as quick relief The short-term treatments are used as quick relief

medicationsmedications• The long-term version is meant to be used regularly to The long-term version is meant to be used regularly to

control the asthma over a long period of timecontrol the asthma over a long period of time

Treatments of Asthma, Treatments of Asthma, Cont’dCont’d

51

• Theophylline has been used for over 30 Theophylline has been used for over 30 years as an asthma treatment. It is taken as years as an asthma treatment. It is taken as either a pill or intravenously. People who either a pill or intravenously. People who use this treatment must have their blood use this treatment must have their blood levels monitored closelylevels monitored closely

• Anticholinergics are always inhaled and can Anticholinergics are always inhaled and can be used with the be used with the -agonist-agonist

Treatments of Asthma, Treatments of Asthma, Cont’dCont’d

52

3.3. CorticosteroidCorticosteroidss

• Better known as steroids, corticosteroids are Better known as steroids, corticosteroids are one of the most effective ways of treating one of the most effective ways of treating asthmaasthma

• They act as an anti-inflammatory, and should They act as an anti-inflammatory, and should not be confused with the anabolic steroids not be confused with the anabolic steroids taken by athletes for muscle enhancementtaken by athletes for muscle enhancement

• Steroids are available in creams, nasal sprays, Steroids are available in creams, nasal sprays, pills, or by injection, and their use must always pills, or by injection, and their use must always be monitored by a doctorbe monitored by a doctor

Treatments of Asthma, Treatments of Asthma, Cont’dCont’d

53

• Oral steroids work best for short-term use Oral steroids work best for short-term use • Long-term use of steroids is best treated by Long-term use of steroids is best treated by

inhaled methods inhaled methods • Long-term use does have lingering effects, Long-term use does have lingering effects,

particularly with oral methods, which include weight particularly with oral methods, which include weight gain, menstrual irregularities, ulcers, high blood gain, menstrual irregularities, ulcers, high blood pressure, weakened bones, and a tendency to bruise pressure, weakened bones, and a tendency to bruise

• Steroid use for children should be done with Steroid use for children should be done with extreme care, as it can stunt their growth. It is extreme care, as it can stunt their growth. It is widely accepted that the benefits of steroid use widely accepted that the benefits of steroid use outweighs the potential side effectsoutweighs the potential side effects

Treatments of Asthma, Treatments of Asthma, Cont’dCont’d

54

THE ENDTHE END

Any questions?