Transcript Case 43 Case Study 2009 Listeria monocytogenes

Case Study
Listeria monocytogenes
2009
Case 43
Roubina Tatavosian
James Muro
Jae Kim
Case Summary
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3 1/2 –weeks old male wan born by Cesarean section
At birth he had a left diaphragmatic hernia, which was repaired soon.
He required intubation and respiratory support.
Over 24-h period, the infant developed following symptoms;
 Bulging anterior fontanelles (A bulging fontanelles is an outward curving of an infant's soft spot )
 Increasing heart and respiratory rate
 Wide fluctuation in blood pressure
 Difficulties maintaining adequate tissue perfusion
 WBC count increased from 6300 to 13700/μl
 Child began to have focal seizures
 Examination of cerebrospinal fluid (CSF) is as following;
 92% neutrophils
 2 mg/dl glucose
 350 mg/dl protein
 3900 WBC/ μl
Key Information Pointing to Diagnosis
• Symptoms of the patient indicates that he had bacterial cause neonatal
meningitis.
• There are two major kind of bacteria that my cause neonatal meningitis;
Listeria monocytogenes and Streptococcus agalactiae in new born.
 Patient was born by Cesarean section not a vaginal delivery so this will eliminate
Streptococcus agalactiae (exist in vagina canal).
 Meningitis occur in the patient after he was born (Late onset kind of meningitis)
 Patient required intubation and respiratory support
 Results of CSF test (92% neutrophils,2 mg/dl glucose, 350 mg/dl protein, and 3900
WBC/ μl )
• Characteristics of CSF Gram Stain; Gram positive rod
• Blood test results
• Isolated colonies from CSF and Gram stain of the male infant proof of
existence of Listeria monocytogenes
Listeria monocytogenes Gram Stain on CSF
The Diagnosis for Case # 43
• This patient was diagnosed by bacterial neonatal meningitis,
caused by Listeria monocytogenes .
• In neonates, neonatal meningitis occur in one of the two forms
(Dr McQueen, Lec note);
– Early onset : Disease acquired in utero;
The infant is infected transplacentally with the production of
septicemia and granulomatous foci in many organs which may
cause , stillbirth, premature delivery, or death soon after birth.
• The baby is born with cardio and respiratory distress, vomiting,
diarrhea, and skin lesions.
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The Diagnosis for Case # 43
• Late onset : Infection occur at or soon after birth.
“In this case study patient was infected with Late
onset neonatal meningitis”
 Infection usually begins 1-4 weeks after birth and is
manifested as meningitis with a high fatality rate.
Classification, Gram Stain Results, and
Microscopic Appearance of Listeria monocytogenes
• Listeria monocytogenes belongs to genus Listeria, Family
Listeriaceae.
• Differential characteristics;
 Gram positive, non-spore forming, motile, facultative anaerobic, βhemolytic, coccobacillus bacterium (Micro lab book) .
• Cultural characteristics;
 Colonies are small, round, smooth, and translucent
 Cell may be found singly, in short chain, or in palisades.
Pathogenesis of L. monocytogenes
• L. monocytogenes is an important human
pathogen
• It’s wide separated in environment (soil, water,
vegetation, and animal products)
• Virulence factor(Mahon, 2007):
Hemolysin (listeriolysin O
Catalase
Superoxide dismutase
Phospholipase C
Surface protein (P 60)
Diseases and Pathogenesis of Disease
Caused by L. monocytogenes
• Listeria monocytogenes causes listeriosis,
septicemia, meningitis, infections in pregnant women,
which may result in spontaneous abortion (2nd/3rd
trimester) or stillbirth.
• In immunosuppressed and older adults, and patient
receiving chemotherapy cause invasive listeriosis.
• In adults symptoms start as mild flu or GI distress.
Diagnosis of Listeria monocytogenes
• In direct CSF smear, L. monocytogenes appear Grampositive
• Grow well in SBA and Chocolate agar (1-2 days).
• Prefer slightly CO2 condition for isolation
• Colonies are surrounded by narrow zone of βhemolysis
Identification of Listeria
monocytogenes
• Isolation of colonies from cerebrospinal fluid (CSF), blood, or
swabs of lesions.
• Biochemical Characteristics (Mahon, 2007);
 Catalase +
 Esculin Hydrolysis +
 Motility +
 Growth in NaCl +
 Hippurate +
 CAMP +
• Streptococcus agalactiae is very similar to
• L. monocytogenes;
 Differentiation:
 S. agalactiae; Catalase -, Motility -
 L. monocytogenes; Catalase + , Motility +
Isolation of Listeria monocytogenes
• Optimal growth temperature is 30˚C to 35 ˚C
• Can grow in the wide range of 0.5 ˚C to 45˚C
• Cold enrichment technique may use to isolate
organism from clinical specimen. (Inoculation of
the specimen into broth and incubation at 4 ˚C for
several weeks).
Therapy, Prevention and Prognosis of Patient
Infected with Pathogen L. monocytogenes
• Therapy for L. monocytogenes;
Ampicillin individually or combined with
Gentamicin,
 Penicillin G individually or combined with
Gentamicin (Micro Lab book, 2005)
Vaccine?
• Vaccine for Listeria monocytogenes is under
development.
Primary Research Article Contributing to the
Understanding of the Disease caused by Listeria
monocytogenes
• S.A.A. Jassim et al, March 2005, The attachment
efficiency of cell-walled and L-forms of Listeria
monocytogenes to stainless steel, Agriculture &
Environment ,Vol.3 (2) : 9 2 - 9 5.
• “Department of Food Science, University of Guelph, Guelph, Ontario, Canada”
Primary Research Article Contributing to the
Understanding of the Disease caused by Listeria
monocytogenes
Purpose:
• To detect the attachment efficiency of cell walled of
L-form of Listeria monocytogenes to stainless steel
surfaces.
Material and method:
• L-form phenotype of Listeria monocytogenes was
induced, propagated and recovered in both broth and
plate culture by exposure to sub-lethal concentrations
of Ampicillin.
Primary Research Article Contributing to the
Understanding of the Disease caused by Listeria
monocytogenes
Results of experiment:
• L-form cells had the capacity to attach well to
stainless steel (0.3% of cells attached) with a higher
efficiency than parental cells (0.002%) after 8 h
exposure to culture.
• After 18 h, the parental cells attached with slightly
higher efficiency (0.8%) than L-forms cells (0.625%).
Primary Research Article Contributing to the Understanding of
the Disease caused by Listeria monocytogenes
Figure 1. L-form colonies of L.
monocytogenes visible after 72 h
on TSA supplemented with 0.5 ng/ml
ampicillin
Figure 2. Scanning electron micrographs of classical and L-forms of L.
monocytogenes. (2a) classical cells with
flagella (arrows), Mag: 10,200x; (2b) L-form cell, Mag: 30,000x.
Primary Research Article Contributing to the
Understanding of the Disease caused by Listeria
monocytogenes
Conclusion:
• The ability of Listeria monocytogenes L-forms to
attach to stainless steel may suggest that a classical
rigid cell wall structure is not a requirement for cell
adhesion in vitro.
• This article relates and supports my case about the
virulence factor Bacterial adhesion of L.
monocytogenes.
Take Home Message
• Meningitis is an inflammation of the meninges, the lining
surrounding the brain and spinal cord
• Typical symptoms are:
Headache, neck stiffness , back stiffness, focal seizures,
nausea, fever, and bulging fontanelles (soft spot on an
infant's head).
• Bacteria cause disease:
Listeria monocytogenes causes meningitis
• Diagnostics:
Cerebrospinal fluid (CSF) test, and blood test.
• Therapy is based on:
Patient age, the organism that cause meningitis, and extinct
on the disease.
Take Home Message
• Prognosis:
 Is poor in neonates; infected mother should be treated as soon as
disease is diagnosed
• Prevention :
 Food safety (cook all food from animal sources, wash raw
vegetables very well, and avoid eating or drinking unpasteurized
milk products).
• Transmission:
 From infected pregnant mother to fetus in utero (early onset)
 Infected from the genital tract during delivery (late onset)
• Treatment:
 Ampicillin, Penicillin
References
• Connie R. Mahon, Donald C. Lehman, George Manuselis. Diagnostic
Microbiology; 3rd ed.;Saunders, an imprint of Elsevier Inc.:2006; Chapter
16.
• Michael J. Leboffe, Burton E. Pierce. A Photographic Atlas for the
Microbiology Laboratry; 3rd ed. Morton Publishing Company, 2005; page
144,153.
• S.A.A. Jassim et al, March 2005, The attachment efficiency of cell-walled
and L-forms of Listeria monocytogenes to stainless steel, Agriculture &
Environment ,Vol.3 (2) : 9 2 - 9 5.
• http://www.aafp.org/afp/990515ap/2761.html
• http://www.cfsph.iastate.edu/Factsheets/pdfs/listeriosis.pdf
• http://www.son.org.tw/db/Jour/2/199806/2.pdf