1. Hepatopancreatic Necrosis Disease (AHPND)-like disease in farmed P
Hepatopancreatic Necrosis Disease (AHPND)-like disease in farmed P. monodon in Queensland
AUSTRALIAN ANIMAL HEALTH LABORATORY (AAHL)
Nick Moody | Team Leader, Aquatic Diagnostic Capability
Ridley Australian Prawn & Barramundi Farmers Symposium – 2 August 2016

2. AHPND Also known as Early Mortality Syndrome (EMS)
Notifiable to the OIE, from 1 January 2016
Led to significant reductions in prawn production globally
Caused by “specific virulent strains of Vibrio parahaemolyticus, namely VPAHPND, which contains one or more extrachromosomal plasmids, including a unique, previously unreported, large, plasmid with a size of ~70 kbp”
Australian disease does not meet the current OIE definition of a confirmed case
Not in V. parahaemolyticus and in one case not in the plasmid
AHPND-like disease in farmed P. monodon in queensland

3. Molecular tests at AFDL
Primer/probe
Sequence (5’-3’)
Target
Amplicon Size (bp)
Reference
VpPirA F
TTGGACTGTCGAACCAAACG
PirA gene
135
Han et al (2015)
VpPirA R
GCACCCCATTGGTATTGAATG
VpPirA probe
FAM- AGACAGCAAACATACACCTATCATCCCGGA-TAMRA
AP1 F
CCTTGGGTGTGCTTAGAGGATG
pVA1 plasmid
700
Flegel and Lo (2014)
AP1 R
GCAAACTATCGCGCAGAACAC C
AP2 F
TCACCCGAATGCTCGCTTGTGG
AP2 R
CGTCGCTACTGTCTAGCTGAAG
AP3 F
ATGAGTAACAATATAAAACATGAAAC
333
Sirikharin et al (2014)
AP3 R
GTGGTAATAGATTGTACAGAA
AP4 F1
PirA + PirB gene
1269
Sritunyalucksana et al (2015)
AP4 R1
ACGATTTCGACGTTCCCCAA
AP4 F2
TTGAGAATACGGGACGTGGG
230
AP4 R2
GTTAGTCATGTGAGCACCTTC
143F
TGCCTTATCAGCTNTCGATTGTAG
18S rRNA gene
848
Lo et al (1996)
143R
TTCAGNTTTGCAACCATACTTCCC
16S-27f
AGAGTTTGATCMTGGCTCAG
16S rRNA gene
1465
Jiang et al (2006)
16S-1492r
AAGGAGGTGATCCAGCCGCA
Vpara toxR-F
GTCTTCTGACGCAATCGTTG
Tox R gene
367
Kim at al (1999)
Vpara toxR-R
ATACGAGTGGTTGCTGTCATG
AHPND-like disease in farmed P. monodon in queensland

4. FRDC : “Aquatic Animal Health and Biosecurity Subprogram: Comparative pathogenicity of exotic AHPND and the presumptive bacterial hepatopancreatitis detected in farmed Penaeus monodon in Queensland
AHPND-like disease in farmed P. monodon in queensland

5. FRDC 2016-013 PI: Nick Moody, AFDL, Geelong
CI: Ian Anderson, BSL, Brisbane
Start Date: End Date:
FRDC funds: $131,106
AHPND-like disease in farmed P. monodon in queensland

6. FRDC 2016-013 Two Australian representative isolates:
North Queensland: PirA+PirB toxin gene in the chromosome of Vibrio harveyi (no pVA1 plasmid present)
Central Queensland: PirA+PirB toxin gene present in the pVA1 plasmid in something other than Vibrio parahaemolyticus (not V. harveyi)
Exotic AHPND strains
PirA+PirB toxin gene in the pVA1 plasmid in V. parahaemolyticus
AHPND-like disease in farmed P. monodon in queensland

7. FRDC
Infectivity trials will be based on the model of Tran et al (2013) included in the current revision of the OIE Manual for AHPND
Use P. monodon as a positive infection control
Use P. merguiensis as a negative extraction control
Whole genome sequence on the PacBio platform (U Melb)
Current MiSeq data generated at AAHL will assist with assembly
Develop material for inter-laboratory test evaluation
Seeking prawns for bioassay trials in August/September 2016
AHPND-like disease in farmed P. monodon in queensland

8. Acknowledgements APFA, Helen Jenkins FRDC AAHBS FRDC QRAC
Biological Sciences Laboratory
CSIRO AAHL
AAHL Fish Diseases Laboratory
AHPND-like disease in farmed P. monodon in queensland

9. Thank you Nick Moody Team Leader, Aquatic Diagnostic Capability
Senior Research Scientist t e
AUSTRALIAN ANIMAL HEALTH LABORATORY (AAHL)

10. Toxin gene location in the Plasmid
AHPND-like disease in farmed P. monodon in queensland