Sabrina Jedlicka

In collaboration with:Prof. Kristen Jellison, Prof. Frank Zhang, Dr. Xia Luo,

Tooba Sarkhosh, Elissa Mayerberger, Lu Luo, and Graeme Nichols

Flagellates:• Giardia lamblia• Dientamoeba fragilis• Chilomastix mesnili• Trichomonas hominis• Enteromonas hominis• Retortamonas intestinalisAmeba:• Entamoeba histolytica• Entamoeba dispar• Entamoeba coli• Entamoeba hartmanni• Endolimax nana• Iodamoeba bütschliiApicomplexa:• Cryptosporidium hominis• Cryptosporidium parvum• Cyclospora cayetanensis• Isospora belliOther:• Blastocystis hominis• Balantidium coli

Intestinal Protozoaunicellular eukaryotic organisms

Source: MMWR – Summary of Notifiable Diseases –

U.S., 2011

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2716703/

CryptosporidiumLife Cycle • Infectious form = oocyst

• Sporozoites ‘invade’

intestinal epithelial cells• Merogony

• produce merozoites• Gametogony

• produce micro- and macrogametes

• Sporogony• produce sporozoites• completed on host cell• thin (autoinfection) or

thick walled oocysts

Image Credits: http://www.hominis.mic.vcu.edu/Cparvum_sporozoites.html; http://www.esemag.com/archive/0199/crypto.htm; Current & Blagburn, 1990

Ingested

Exits Host

OocystSporozoite Type I Meront

Merozoite

Type II Meront

MerozoitesMicrogamont

Macrogamont

Zygote

Thin-walledOocysts

Thick-walledOocysts

Auto-infection

Host Enterocytes

SexualCycle

1. Filtration

2. Elution

3. Immunomagnetic Separation (IMS)

4. Immunofluorescent Assay (IFA)

(Helmi et al. 2008; Howe et al. 2002; Rogers & Keevil 1995; Searcy et al. 2006; Wolyniak-DiCesare et al. 2012; Wolyniak et al. 2009; Wolyniak et al. 2010)

Mature biofilm at different shear

Mature biofilm by season

http://www.marvistavet.com/assets/images/single_cryptosporidium_oocyst.gif

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1) Little lehigh River - Light

Neq=217.58 oos/cm2

Kd=0.12hr-1

R2=0.80

# oo

cyst

s/un

it su

rfac

e ar

ea (

oos/

cm2 ) 2) Little lehigh River - Dark

Neq=260.04 oos/cm2

Kd=0.017hr-1

R2=0.80

Time (hr)Time (hr)

# oo

cyst

s/un

it su

rfac

e ar

ea (

oos/

cm2 )

Neq= 214.3 oos/cm2

Kd=0.087hr-1

R2=0.86

3) Monocacy Creek - Light 4) Monocacy Creek - Dark

Neq=232.9 oos/cm2

Kd=0.027hr-1

R2=0.97

Conceptual model of oocyst deposition onto biofilm due to calcium bridging with carboxylic groups.

Magnesium may form an outer sphere surface complex with oocysts and biofilm surfaces to slightly enhance oocyst deposition by charge neutralization.

Cryptosporidium Oocyst Surface Properties

Force Spectroscopy to Measure Binding Forces

Study of oocyst adhesion force to different surface models

(hydroxyl, amine and carboxyl) with atomic force microscopy

(AFM) method.

A powerful tool to measure the mechanical properties of macromolecules, and to investigate the forces and motions

associated with molecular activities at single molecule level

Schematic drawing of AFM operation

Approach

Contact

Retraction

Separation

oocyst

Surfacemodel

AFMprobe

Force curves

Detachment(rupture)force

Approach

Contact

Retraction

Separation

Three sample AFM curves showing theinteraction between oocysts andmodel surfaces

1.Nointeraction

2.Intermediateruptureforce

3.Strongestadhesion

*

0.00E+00

1.00E-09

2.00E-09

3.00E-09

4.00E-09

5.00E-09

6.00E-09

7.00E-09

week1 week2 week3 week4 week5 week6 week7 week8

Force(N)

OH

NH2

COOH

*

*

**

*

*

*

*

*

** *

**

*

#

#

#

#

#

#

#

#

#

#

#

^

^

^^

^

^^

^

0.00E+00

2.00E-10

4.00E-10

6.00E-10

8.00E-10

1.00E-09

1.20E-09

week 1 week 2 week 3 week 4 week 5 week 6 week 7 week 8

Force(N)

OH

NH2

COOH***

##

With Divalent Cations Without Divalent Cations

0.00E+00

2.00E-10

4.00E-10

6.00E-10

8.00E-10

1.00E-09

1.20E-09

week1 week2 week3 week4 week5 week6 week7 week8

Force(N)

without divalent ions

with divalent ions

0.00E+00

1.00E-09

2.00E-09

3.00E-09

4.00E-09

5.00E-09

6.00E-09

7.00E-09

8.00E-09

9.00E-09

week1 week2 week3 week4 week5 week6 week7 week8

Force(N)

without divalent ions

with divalent ions

0.00E+00

5.00E-10

1.00E-09

1.50E-09

2.00E-09

2.50E-09

3.00E-09

week1 week2 week3 week4 week5 week6 week7 week8

Force(N)

without divalent ions

with divalent ions

*

*

**

*

*

*

*

*

*

**

*

*

*

(a)

(b)

(c)

(d)

https://pubs.rsc.org/en/content/articlelanding/2015/py/c4py01415d/unauth#!divAbstract

Hyaluronic Acid, Chondroitin Sulfate, Heparin, Many others

kjellison@lehigh.edu

ssj207@lehigh.edu