MLS Student presentation on Cryptosporidium

1. cdc.gov/parasites/crypto/
Angie Bain
Stephanie Hood

2.  Protozoan
 Phylum: Apicomplexa
 Class: Sporozoasida
 Order: Eucoccidiida
 Family Cryptosporiidae
 Genus: Criptosporidium waterfilterreview.com
 Species: parvum, muris, meleagridis, felis, etc.

3.  Recognized in mice in 1907
 Reported in humans in 1976
 Immunocompetent child
 Immunosuppressed adult
 Recognized globally in 1980s
and 1990s
www.nap.edu
 AIDS patients
Ernest Edward Tyzzer
 Outbreak among veterinary
students

4.  Cryptosporidium is a
spore producing parasite
found in the intestine of
infected people and
animals.
 Cryptosporidium spp. is landesbioscience.com
the most common cause
of Cryptosporidiosis.

5.  Infection
 Ingesting food or drinks
contaminated with fecal
material
 Swallowing recreational
water contaminated with secretsofsoa.com
Cryptosporidium
 Not washing hands
 Sexual practices leading
to oral exposure with
fecal material
victorystore.com

6.  Buries into intestinal
lining of the gut
 Goes through Life Cycle
 Alters osmotic pressure
 Diarrhea
 Atrophy of intestinal villi
 Alters uptake of fluids,
electrolytes, and nutrients
 Malabsorption syndrome
CDC DPDx Library

7.  Symptoms (2 to 10 days after
infection and can last up to
30 days)
 Diarrhea
 Stomach cramps treehugger.com
 Dehydration
 Nausea
 Vomiting
 Fever
 Weight loss
 Sometimes no symptoms are
seen
bathroomscalereview.com

8.  White Blood Cells
Phagocytize Parasites
 Segmented neutrophils
 Macrophages
 Lymphocytes
 Eosinophils
labmed.hallym.ac.kr

9.  CD4+ T cells
 Early infection
 CD8+ T cells
 Elimination
 CD154 and CD40
 Stimulate nitric oxide
 IFN-γ, IL-12
 T cell response
 Apoptosis
 Other Cytokines
 TNF-α, IL-1β, IL-2, IL-
4, IL-10, IL-15, etc.
 Patients with AIDS
 Decreased CD4+ count Ashton-Rickardt, 2004

10.  IgM
 IgG
 IgA
 X-linked
immunodefiencey
 Mutations in CD154
gene
 Defected IgM cannot allerresponz.com
mount immune
response

11.  Specimen Source
 Multiple Stool Specimens
 Diagnostic Techniques
 Wet Mount
 Modified Acid Fast Stain
 Direct Fluorescent Antibody (DFA)
Assay
 Detection Methods
 Safranin Stain
 Trichrome Stain
 Enzyme Immunoassay (EIA) savethecanyoncall.com
 Polymerase Chain Reaction (PCR)
 Rapid Immunochromatographic
cartridge Assays

12.  Visualization of Oocysts
 4 to 6 µm
 Bight-Field Microscopy
 Differential Interference
contrast (DIC)
 High Sensitivity and CDC DPDx Library
Specificity Wet Mount Slide

13.  Visualization of Oocytes
 Light pink to dark red
 Can also visualize sporozoites
 Relatively High Sensitivity
and Specificity
 Irregular Staining
CDC DPDx Library
 cause “ghost” oocysts Modified Acid Fast Stain

14.  Fluorescence microscope
 “Gold Standard”
 High sensitivity and
specificity
 Does not provide
archievable stained slide
 Requires special equipment CDC DPDx Library
Fluorescent Stain
Auramine Rhodamine

15.  Safranin stain
 Oocysts stain a bright red
orange
 Not widely used because
oocysts may not stain properly
Trichrome Stain
CDC DPDx Library

Safranin Stain
 Oocysts may appear unstained
 Lowest sensitivity and
specificity among all tests
 Can detect Oocysts, but
Cryptosporidium should be
confirmed by diagnostic CDC DPDx Library
techniques Trichrome Stain

16.  Detects isolated antigens
from a patients sample
using antibodies that are
tagged with a color
changing enzyme.
 Relatively high
Sensitivity and
Specificity
 Does not involve
microscopy techlab.com
 Screens large numbers of
specimens

17.  Separates DNA
fragments based on
size
 435 bp
 High Sensitivity and
Specificity
CDC DPDx Library

18.  Detects isolated antigens
from sample using
antibodies. A positive test
is indicated by a colored
bar.
Variable Sensitivity and
alibaba.com

Specificity
 Some Assays Have Been
Recalled

19.  Nitazoxanide
 Paromomycin
 Azithroycin
 Individuals with
AIDS
 anti-retroviral therapy
generecsmed.com

20.  Ashton-Rickardt, P.G. A license to remember. Nat. Immunol.
5, 1097-1098 (2004).
 Beach, M.J. And Johnston, S. P. Manufacturer's recall of rapid
cartridge assay kits on the basis of false-positive Cryptosporidium
antigen tests — Colorado. MMWR. 53, 198-199 (2004).
 CDC. http://www.cdc.gov/parasites/crypto/. Accessed
September 22, 2011
 Dillingham, R. A., Aldo, A. A., and Guerrant, R. L.
Cryptosporidiosis: epidemiology and impact. Microb. Infect.
4, 1059-1066 (2002).
 Mahon, C.R., Lehman, D.C., and Manuselis, G. Textbook of
Diagnostic Microbiology. 4th Ed. Elsevier (2011)
 Riggs, M. W. Recent Advances in cryptosporidiosis: the immune
response. Microb. Infect. 4, 1067-1080 (2002).
 Stevens, C.D. Clinical Immunology & Serology: A Laboratory
Perspective. 3rd Ed. F.A. Davis Company (2010).
 Tzipori, S. and Honorine, W. Cryptosporidiosis:
biology, pathogenesis and disease. Microb. Infect. 4, 1047-1058
(2002).