1. No category

ihbi
cells and t issue domain
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Chlamydia and koalas : a
battle to be won .... or lost
Range of infections & disease caused by
Chlamydia in koalas
 Inapparent / sub-clinical
 Ocular
– Mild conjunctivitis
– Complete blindness
 Urogenital
– Bladder infections, cystitis, cysts, prostatitis
 Respiratory
– Rhinitis, sneezing, cough, pneumonia
Chlamydial infections in koalas
Cystitis
“Wet bottom”
Reproductive tract
Cysts “Mars Bar”
Conjunctivitis
“Blindness”
Chlamydial developmental cycle
Stress:
- Penicillin
- Heat
- IFN – γ etc
Normal inclusion
Persistence
Aberrant
body
(Polkinghorne et al., 2006)
(Belland et al., 2003)
Old Taxonomy
New Taxonomy
Current Taxonomy
(Everett et al., 1999)
Family
Chlamydiaceae
Family
Chlamydiaceae
Genus and species
Chlamydia
 C. trachomatis
 C. psittaci
 C. pecorum
 C. pneumoniae
Genus and species
Chlamydia
 C. trachomatis
 C. suis
 C. muridarum
Chlamydophila
 Cp. psittaci
 Cp. abortus
 Cp. caviae
 Cp. pecorum
 Cp. pneumoniae
 Cp. felis
(Stephens et al., 2009)
Family
Chlamydiaceae
•
•
•
•
•
•
•
•
•
Genus and species
Chlamydia
C. trachomatis
C. suis
C. muridarum
C. psittaci
C. abortus
C. caviae
C. pecorum
C. pneumoniae
C. felis
Chlamydia testing methods





Clinical signs : eg. conjunctivitis, wet bottom
Cell culture : very specialised
Antibody detection : serology; ELISA
Antigen detection : Clearview test kit
DNA probes
 PCR / qPCR
Epizootiology of chlamydial infection in Australia’s
wild koala population
Summary
Population
% +ve
Magnetic Island
Narangba
Mutdapilly
Koala Coast
Coombabah
East Coomera
Koala Beach
Port Macquarie
Pilliga State Forest
Pine Creek State Forest
Ballarat
French Island
Mt Lofty Ranges
Kangaroo Island
0%
50%
85%
87%
10%
35%
67%
45%
12%
72%
100%
0%
90%
0%
Summary
Population
% +ve
C.pecorum
C.pneumoniae
Magnetic Island
Narangba
Mutdapilly
Koala Coast
Coombabah
East Coomera
Koala Beach
Port Macquarie
Pilliga State Forest
Pine Creek State Forest
Ballarat
French Island
Mt Lofty Ranges
Kangaroo Island
0%
50%
85%
87%
10%
35%
67%
45%
12%
72%
100%
0%
90%
0%
0%
50%
73%
87%
10%
35%
67%
38%
8%
52%
90%
0%
90%
0%
0%
21%
4%
10%
10%
23%
8%
12%
20%
0%
53%
0%
C.pecorum in koalas
1. What is the diversity of C.pecorum
infections in koalas ?
2. Where did koalas get their C.pecorum
from ?
What is the origin of C. pecorum in koalas?
Ancient USA / Europe / Asia
birds / animals
strains
Australia (1788)
Phylogenetically diverse
sheep / cattle strains
C. pecorum line
Range of hosts
Major outer membrane protein (ompA)
 ompA
– MOMP
 Important functions:
– Porin
– Attachment
– Structural stability
(Kaltenboeck et al., 1993)
 CDs and VDs
– Target of molecular studies
Kaltenboeck et al.,
1993
Kaltenboeck et al.,
1993
C.pecorum phylogeny using the ompA gene only
C.pecorum phylogeny using several genes
ompA, incA, ORF663
ompA
C.pneumoniae infections in koalas
1. Is C.pneumoniae significant in koalas ?
2. How does the koala strain of
C.pneumoniae relate to the human
strains of C.pneumoniae ?
1999
Chlamydia much more widespread than first
thought
 Berger ……….. & Timms (1999)
 C.pneumoniae identified in a free-ranging Giant Barred frog
 Gene sequences extremely similar to koala C.pneumoniae
strains
Koala C.pneumoniae – strain LPCoLN
 Respiratory disease outbreak in a large captive koala colony
– 70% of animal affected over 6-9 month period
– Rhinitis …. Coughing/sneezing ….. Serous/purulent discharge
– spread through the colony via respiratory route
 No other bacteria or viruses isolated
 Chlamydia / C.pneumoniae diagnosed via antigen detection and
PCR
 Nasal swabs from Connor cultured in Hep2 cells at 37C →
LPCoLN
Host range of C.pneumoniae
 Humans
 Amphibians
 Horses (Storey et al., 1993)
 Marsupials
– koalas (Jackson et al., 1997;
Wardrop et al., 1998)
– bandicoots
(Bodetti et al.,
2002)
– gliders
– frogs
(Berger et al., 2000; Reed
et al., 2000; Hotzel et al., 2001;
Blumer et al., 2007)
 Reptiles
– snakes
(Bodetti et al., 2002;
Jacobsen et al., 2004)
– crocodiles
(Bodetti et al., 2002)
– iguanas
(Bodetti et al., 2002)
Current status of C.pneumoniae evolution
Some recent epidemiology
lessons from SE Queensland
populations
Populations analysed
 Narangba (A)
 Brendale (B)
 East Coomera (C)
Epizootiology
Population
q PCR +ve
Animals*
Male/Female
qPCR +ve
<1yr
1-5yrs
>5yrs
Population A
63 swabs from 16
animals
8 (50%)
3:5
1/8
1/8
6/8
Population B
83 swabs from 22
animals
11 (50%)
5:6
0/11
3/11
8/11
Population C
51 swabs from 17
animals
6 (35%)
4:1
0/5
2/5
3/5
•1 = < 10 copies/ul; 2 = 10-100 copies/ul; 3 = 100-1000 copies/ul; 4 = 1000-10000 copies/ul; 5 = >10000 copies/ul
Genetic diversity of C.pecorum infections
CpeNTVD3
VD1
VD2
CpeNTVD4
VD3
VD4
331bp
MOMP
Genotype
A
B
C
D
E
E'
F
G
H
Sample
LPDowellV495
MT113V189
MT213V189
RBNarO1095
CS9745V496
CS9762V496
PMKalaO594
RI28Vna
FWPBilbPna
K4
K13
EMDuarV496
PMRiorV295
PMScarVna
PMBossPna
Nar/Edna/UGT
Nar/Dion/L/E
Nar/Dion/UGT
Nar/Frankie/UGT
Nar/Felix/UGT
Nar/Greg/Nasal
Nar/Kaia/UGT
Bre/Pnau/UGT
Bre/Kurt/UGT
Bre/Poppy/UGT
Bre/Michael/UGT
Bre/Xena/UGT
Bre/Nigel/UGT
EC/Ned/L/E
EC/Ned/R/E
EC/Graeme/UGT
Bre/Miss Radio/UGT
EC/Steve/UGT
EC/James/UGT
Mars Bar/UGT
EC/Ned/UGT
EC/Steve/R/E
Geographical location
Lone Pine Koala Sanctuary, QLD
Mutdapilly, QLD
Mutdapilly, QLD
Redland Bay, QLD
Currumbin Sanctuary, QLD
Currumbin Sanctuary, QLD
Port Macquarie, NSW
Raymond Island, VIC
Featherdale Wildlife Park, NSW
Australia
Australia
Emerald, QLD
Port Macquarie, NSW
Port Macquarie, NSW
Port Macquarie, NSW
Population A
Year of Study
Jackson et al., 1997
Jackson et al., 1997
Jackson et al., 1997
Jackson et al., 1997
Jackson et al., 1997
Current Study
Population B
Current Study
Population C
Population B
Population C
Population C
Mt.Cotton, Brisbane
Current Study
Population C
Current Study
Diversity of MOMP genotypes
F
G
F
G
F
G
H
Population B Population C
E’
Population A
SEQ Map
OmpA genotypes of koala C. pecorum isolates
Genotypes A,B,C and D identified in
1999 by Jackson et al. Vet Micro
Genotypes E,E’,F,G and H identified
in 3 study populations
Collaborators






QUT
Peter Timms
Ken Beagley
Alison Carey
Adam Polkinghorne
Avinash Kollipara










Lone Pine Koala Sanctuary
Galit, Jacqui & Karen
Australian Wildlife Hospital
Jon & Jo
Gold Coast City Council
John & team
Pfizer Animal Health
John Walker
Friends of the Koala (Lismore)
Lorraine Vaas

Supported by ARC, Qld Gov