Diagnostic microbiology lecture: 15 Bordetella pertussis Abed ElKader Elottol MSc. Microbiology - PowerPoint PPT Presentation

Diagnostic microbiology lecture: 15 Bordetella pertussis Abed ElKader Elottol MSc. Microbiology 2010.

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Diagnostic microbiologylecture: 15Bordetella pertussis

Abed ElKader Elottol MSc. Microbiology

2010

Abed ElKader ElOttol

Abed ElKader ElOttolAbed ElKader ElOttol

Disease: whooping cough (Pertussis)

  • Pertussis = Transmitted by the respiratory tract rout from early case and possibly via carriers.
  • The organism adheres to and multiply rapidly on the surface of the epithelium in the trachea and bronchi and interfere with ciliary action .
  • Disintegrating organisms liberate a toxin that irritate surface cells, giving rise to catarrhal symptoms.

Abed ElKader ElOttol

An acute respiratory disease with three stages:

A catarrhal stage with an irritating cough, lasts 1 to 2 weeks.

A paroxysmal stage characterized by violent coughs followed by a high respiratory whoop, lasts 2 to 6 weeks.

A convalescent stage where the cough gradually decreases in frequency and severity, lasts several weeks.

Note: 75% of deaths are among infants.

Signs & Symptoms of Pertussis

  • The first symptoms of pertussis are similar to those of a common cold:
  • runny nose
  • sneezing
  • mild cough
  • low-grade fever
  • Most B.pertussis strains contains peptide that promotes marked lymophocytosis in the host.
  • The toxin is heat stable.
    • Elevated white blood cell count with a lymphocytosis, Confirmed with laboratory testing.
Abed ElKader ElOttolAbed ElKader ElOttol

Organism:

  • Short, ovoid, gram-negative bacilli.
  • Aerobic.
  • Complex, enriched media is required for isolation (Bordet-Gengous).
  • Non motile, urease negative, nitrate negative.
  • Laboratory Diagnosis:
  • Nasopharyngeal swabs or cough droplets, expelled onto "Cough plates" held in front of the mouth of the patient during a paroxysms.
  • Incubate (Bordet-gengou`s plates) at 35 oC for 2-5 days.
  • Typical colonies resemble a 1mm droplets of mercury surrounded by a zone of hemolysis.
  • Colonies are confirmed with specific antiserum Direct immunofluorescent staining of smears made from nasopharyngeal swabs may give a rapid positive test.

Abed ElKader ElOttol

Human Disease & Associated Pathogens

Abed ElKader ElOttolAbed ElKader ElOttol

Legionella pneumophila

Abed ElKader ElOttol

Legionellosis was first recognized in association with the epidemic of legionnaires disease which occurred in Philadelphia in July 1976.

  • Shortly following the epidemic, the causative agent was isolated from lung tissues of patient who died of this disease.
  • The organism was named Legionella pneumophila.

Abed ElKader ElOttol

ORGANISM:

Gram negative rods.

Stain poorly with gram stain unless safranin is applied for long periods.

Slow grower (2-5 days in enriched media containing L-cystein and supplemented source of ferric iron).

Old culture may exhibit filamentous forms, swollen rods, and bizarre forms.

Soluble pigment that results in brown coloration on Feely-Gorman agar.

Good growth obtained on Charcoal-Yeast Extract (CYE).

Good growth occur in an atmosphere containing 2.5% CO2.

Most strains are flagellated.

Abed ElKader ElOttol

Laboratory Diagnosis
  • Culture gold standard
    • Require L-cysteine and iron salts
    • Use BCYE (buffered charcoal yeast extract agar)
    • Antibiotics might be used to help inhibit the growth of fastidious bacteria
    • Grown aerobically or
    • Grow using 3-5% CO2 at 35° for 3 to 5 days
    • pH 6.9 for optimum growth
    • Colonies are small with a crystalline-like, ground glass appearance
Abed ElKader ElOttolAbed ElKader ElOttol

Biochemical Characteristics:

  • Weak oxidase positive
  • Strong catalase positive
  • Liquefy gelatine
  • No carbohydrate fermentation
  • No nitrate reduction
  • No urea hydrolysis
  • Hydrolysis of hippurate Positive (This differentiate it from other legionella species)
  • Virulence factors:
  • 1. Proteolytic enzymes
  • 2. Exotoxin with lysed laboratory animal RBCs
  • 3. Cytotoxin which caused inhibition of cell growth in laboratory animals.

Abed ElKader ElOttol

Clinical Manifestation:

  • Acute pneumonia begins 2-10 days after exposure with a brief prodorm of malaise, myalgia, and headache, followed rapidly by prostration, high fever.
  • an rigors Cough, dyspnea, pleuritic and abdominal pain, vomiting, diarrhea, and unexplained encephalopathy are often seen.
  • Gram stain of sputum is not diagnostic.
  • Leukocytosis, elevated ESR, proteinurea, hematuria and abnormal serum enzyme determination are common.
  • The common causes of death due to respiratory failure and shock.

Abed ElKader ElOttol

Abed ElKader ElOttolAbed ElKader ElOttol

Transmission:

  • Via respiratory tract.
  • Collection & Processing of Speciemn:
  • Caution: Specimen must be handled in a biological safety cabinet.
  • Lung tissue obtained at autopsy or biopsy is optimally selected from areas ofnecrosis.
  • A representative portion of specimen should be placed in 10% neutral formalin for Direct Fluorescent Antibody and histopathological examination.
  • 2. Pleural fluid, transtracheal aspirate.

Abed ElKader ElOttol

Abed ElKader ElOttol

Diagnosis:

1. Isolation and identification of Legionella from clinical specimens either on artificial media or guinea pig inoculation followed by embryonated hen`s egg.

2. Demonstration of a four-fold or greater rise in antibody titer from acute phase to convalescent phase by the IFA test.

3. Demonstration of the organism in clinical specimen by DFA.

Treatment:

Erythromycin is the drug of choice.

Abed ElKader ElOttol

BRUCELLA

Abed ElKader ElOttol

• Gram-negative bacilli

• Chronically infected domestic animals.

Medically Important Species

B. abortus – Cattles

B. melitensis - Goats & sheeps

B. canis – Dogs

B.suis swine

Abed ElKader ElOttol

TRANSMISSION

  • • Primarily animal disease : Causes abortion & sterility.
  • Transmission
  • From animals to animals & humans by:
  • Abrasion in skin
  • Inhalation
  • Ingestion of contaminated milk & cheese
  • Persons at High Risk
  • Dairy workers & farmers
  • Live stock handlers & veternarians
  • °Slaughterhouse employees

Abed ElKader ElOttol

CLINICAL FEATURES OF BRUCELLOSIS (Undulating fever)

• Incubation period: 1-3 weeks

• Initially influenza-like.

Headache, weakness, weight lost, arthralgia, liver dysfunction.

• Undulating (rising & falling) fever for weeks & months.

• A chronic illness

• Splenomegaly

• Hepatomegaly

• Enlarged lymph nodes

Abed ElKader ElOttol

Abed ElKader ElOttol

LAB DIAGNOSIS

• Specimens

Blood, LN & BM biopsy

• Culture

Require 5-10% CO2

• Serology

° Antibody titre of 1:160 or more (significant)

° Titre returns to normal within a year of successful treatment

TREATMENT

• Ciprofloxacin

May be up to 6 weeks to prevent relapses

Abed ElKader ElOttol

PREVENTION

• Pasteurization of milk

• Immunization of animals

• Eradication of infected livestock

• Minimize occupational exposure

Abed ElKader ElOttol

End of Lecture

Abed ElKader ElOttol

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