Comparative biology of Cryptosporidium hominis and the

Cryptosporidium rabbit genotype

Rachel Chalmers1, Guy Robinson1, Steve Wright2, Paul Hunter3, Kristin Elwin1, Maha Bouzid3,
Lee Innes2, Steve Hadfield1, Kevin Tyler3, Frank Katzer2
1UK

Cryptosporidium Reference Unit, NPHS Microbiology Swansea, UK

2Moredun Research Institute, Edinburgh, UK
3University of East Anglia Medical School, Norwich, UK

Background: In June 2008, a waterborne outbreak in Northamptonshire (UK) was caused by the a newly emerging pathogen,
Cryptosporidium rabbit genotype. Twenty-two human cases were confirmed during this outbreak, some with severe
illness. This genotype, previously only identified in five rabbits and a single published human, is closely related to
Cryptosporidium hominis and its taxonomic status is uncertain.
Aims: 1. To study the morphology, genome and experimental host range of the rabbit genotype in comparison with C. hominis.
2. To investigate the taxonomic status of the rabbit genotype.

Method:

Comparative Investigations with C. hominis

Rabbit Genotype
Isolates

Morphology &
Staining
Characteristics

Experimental
Infectivity

Differential Interference
Contrast

Genome Analysis

Commonly investigated
genes:
Small-subunit rRNA,
70kDa Heat Shock Protein,
Actin, COWP

NZW Rabbits

Rabbit (Source)

Modified Ziehl-Neelsen
C. hominis

Baby & Immunosuppressed

Adult Porton Strain Mice
C. parvum

C. parvum

Human cases

Rabbit genotype

C. hominis

Length

5.98 m ( 0.18)

5.35 m ( 0.17)

Width

5.38 m ( 0.24)

4.79 m ( 0.25)

Shape (L:W)

1.11 ( 0.06)

1.12 ( 0.07)

Oocyst features
and staining

Typical

Typical

Cross-reactivity
with IFA (Cellabs)
& IMS Reagents
(Dynal and TCS)

Genome Analysis

Experimental Infection

Rabbit genotype

Ultrastructure of
life-cycle stages

Subtyping gene:
60kDa Glycoprotein
(microsatellite + downstream
sequence)

HCT-8 cells

DAPI

Morphology

Multilocus genome and

hierarchical cluster analysis:
11 genes

Immunosuppressed
Mongolian gerbils

FITC labelled
immunofluorescence

Results:

Currently under
investigation by
electron microscopy

Currently under
investigation by
electron microscopy

Yes

Yes

Anthroponotic

Auramine Phenol

Drinking water

C. hominis

% Difference

NZW Rabbits

Yes
High shedding
by day 6

Very few oocysts

from a single animal
on one day only.

Partial Gene
(bp compared)

Baby Mice

No

No

Immunosuppressed Yes
Adult Mice
High shedding
by day 4

Yes
Low shedding from
day 10

Immunosuppressed Yes
Mongolian Gerbils
High shedding
by day 5

Yes
High shedding from
day 11

HCT-8 Cells

Yes* Infection in
clusters

Yes Infection
in clusters

Shedding patterns were significantly different between

rabbit genotype and C. hominis.
Infected animals showed some pathological changes
but no clinical signs.

Rabbit genotype
vs. C. hominis

Cf. C. hominis
vs. C. parvum

ssu rRNA (785bp)

0.51

1.02

Hsp70 (397bp)

0.25

1.51

Actin (990bp)

0.10

1.51

COWP

0.00

1.38

Multilocus (4469bp)

0.27

1.72

gp60 (822bp)

New subtype family Va

(Outbreak subtype: VaA18)
18-20% different from
C. hominis & C. parvum

Conclusions: There are small, but clear, differences in the morphology, genomes and hierarchical cluster analysis of the rabbit
genotype and C. hominis.
The infection/transmission profiles in the host species studied here are very different.
Until there is further evidence (e.g. from sympatric character displacement studies) it is taxonomically useful to
consider closely related cryptosporidia as subspecies of each other, and we propose C. hominis cuniculus and
C. hominis hominis respectively.
Acknowledgements: This study was funded by Defra, administered by the Drinking Water Inspectorate.

* Hashim et al. (2004) Infect Immun 72(10):6125-6131.