GL BALMICROBI ME SYMPOSIUM 2018 - The Harry Perkins Institute of Medical Research QEII Medical Centre, Nedlands WA - West Coast Microbiome Network
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Proudly presents
GL BAL MICROBI ME
SYMPOSIUM 2018
Bringing together leaders in environmental and clinical
microbiome-related research in WA
August 17th, 2018
The Harry Perkins Institute of Medical Research
QEII Medical Centre, Nedlands WA
Symposium sponsors
Gold
Silver
2
Bronze
Poster prize sponsor
3
Symposium organising committee
A/Prof Christopher Peacock
Prof Andy Whiteley
Prof Thomas Riley
Prof Jeffrey Keelan
Prof Elizabeth Watkin
Dr Andrea Paparini
Dr Claus Christophersen
Dr Daniel Knight
Dr Papanin Putsathit
Dr Matthew Payne
Ms Stacey Hong
Mr Benjamin Moreira Grez
Dr Joshua Ravensdale
Dr Jose Caparros-Martin
Ms Rachael Lappan
Dr Charlotte Oskam
Contact details
http://www.wcmicrobiome.org/
wcmicrobiome@gmail.com
@wcmicrobiome
4
EVENT PROGRAMME
Friday 17th August Harry Perkins Institute of Medical Research
Registration
8:30-9:00
Session I (Chair: Christopher Peacock)
9:00-10:30
Opening Address 9:05-
Barry Marshall (UWA) 9:25
Plenary Presentation - Human microbiome research: Challenges and 9:25-
opportunities 10.05
Geraint Rogers (SAHMRI)
Dispersal limitation and selection pressures as controls on microbial 10.05-
community assembly in engineered environments 10:30
Talitha Santini (UWA/UQ)
Coffee/Tea
Q&A with Barry Marshall and school students
10:30-11:00
Session II Metagenomics in Health and the Environment 1
(Chair: Elizabeth Watkin)
11:00-12:40
The gut microbiome of an ageing-host: friend or foe? 11:00-
Parag Kundu (SCELSE) 11:20
Microbial mats in lakes on Rottnest island 11:20-
Deirdre Gleeson (UWA) 11:40
Profiling the diversity of Cryptosporidium species and genotypes in 11:40-
wastewater treatment plants in Australia using next generation 12:00
sequencing
Alireza Zahedi (Murdoch)
Seeding of the fetal microbiome 12:00-
Lisa Stinson (UWA) 12:20
Long term effects of breastfeeding on immune health by the shaping of 12:20-
gut microbiota 12:40
Valerie Verhasselt (UWA)
5
Lunch and Poster session
12:40-13:30
Session III Metagenomics in Health and the Environment II
(Chair: Jeffrey Keelan)
13:30-15:10
Seasonal dynamics of eukaryotic and prokaryotic diversity in an acid 13:30-
saline lake 13:45
Noor-ul-Huda Ghori (UWA)
Early cystic fibrosis lung disease: effects of the early microenvironment 13:45-
on bacterial populations 14:00
Stephen Stick (UWA/TKI)
The benefits and pitfalls of using amplicon next-generation sequencing 14:00-
to investigate tick microbiomes 14:15
Telleasha Greay (Murdoch)
Using ‘omics technologies to understand the pathogenesis and seek 14:15-
alternative therapies for otitis media in children 14:30
Rachael Lappan (UWA/TKI)
See the microbial world in high resolution 14:30-
James Miller (Millennium Science) 14.50
Flash talks
Phylogenetic and predicted functional profile of microbial communities 14:50-
in biological soil crusts of the Midwest in Western Australia 14:55
Kang Tam (Curtin)
The impact of colostrum on growth 14:55-
Lieke van den Elsen (UWA) 15:00
Targeted amplicon next-generation sequencing for screening and 15:00-
genotyping of pathogens in cattle 15:05
Megan Jordan (DPIRD)
Characterisation of the human milk microbiome from mothers who 15:05-
delivered preterm and term infants 15:10
Ali Sadiq Cheema (UWA)
Flash talk questions
15:10-15:15
6
Coffee/Tea
15:15-15:45
Session IV Translation of the Microbiome
(Chair: Claus Christophersen)
15:45-17:20
Long-term Paleolithic diet is associated with low resistant starch, high 15:45-
saturated fat intake, increased TMAO concentrations and different gut 16:00
microbiota composition
Angela Genoni (ECU)
Echoes of the microbiome: Irritable bowel syndrome diagnosis through 16:00-
bowel sounds 16:15
Mary Webberley (UWA)
Sampling scale, scope and resolution affect interpretation of termite 16:15-
core microbiomes 16:30
Ghislaine Platell (UWA)
Life after death: thanatomicrobiome and its application in forensic 16:30-
science 16:45
Paola Magni (Murdoch)
Isoquercetin and inulin synergistically modulate the gut microbiome to 16:45-
prevent development of the metabolic syndrome in mice fed a high fat 17:00
diet
Natalie Ward (Curtin)
Quality assurance in P4 systems medicine: the impact of preanalytical 17:00-
processes, biobanking protocols and analytical platform stability 17:20
David Broadhurst (ECU)
Poster and flash presentation prizes and wrap-up
17:20-17:30
Sun downer @ Charlies Social Club
17:30-
7
Invited talks
Human microbiome research: challenges and
opportunities
Geraint Rogers1
1South Australian Health and Medical Research Institute, North Terrace, Adelaide SA, Australia
Over the last two decades, the discipline of medical microbiome research has
emerged as an important component of human microbiology. This process, which
has seen techniques and concepts borrowed from the field of environmental
microbiology and applied to the analysis of clinical samples, has fundamentally
changed our understanding of how microbes contribute to disease. It has become
increasingly apparent that interactions with our resident microbial populations shape
virtually every aspect of human physiology. For example, the microbiome influences
the development of our immune systems, the establishment of metabolic control, and
even extends to the regulation of features of our central nervous system. Human
microbiome research is now being focused on addressing healthcare challenges that
increasingly define our longevity and well-being. These include the rapid rise of
cardiometabolic disorders, the ability to deliver effective vaccinations, and the threat
posed by a post-antibiotic era. Meeting these challenges will require us to move
beyond the description of host-microbiome interactions that has been a major feature
of microbiome research to date, and towards translational strategies that enable us
to manage the human microbiome for improved clinical outcomes.
8
Dispersal limitation and selection pressures as controls on
microbial community assembly in engineered
environments
Talitha Santini1,2, Ben J Woodcroft3, Gene Tyson3, Lesley Warren4
1 School of Agriculture and Environment, The University of Western Australia, Crawley WA, Australia
2 School of Earth and Environmental Sciences, The University of Queensland, Brisbane QLD, Australia
3 Australian Centre for Ecogenomics, School of Chemistry and Molecular Biosciences, The University of
Queensland, Brisbane QLD, Australia
4 Lassonde Institute of Mining, Department of Civil and Mineral Engineering, University of Toronto, Toronto ON
M5S 1A4, Canada
Microbial community succession in tailings materials produced by minerals and
energy industries is currently poorly understood, and likely to be substantially
different from similar processes in natural primary successional environments due to
the unusual geochemical properties of tailings and the isolated design of tailings
storage facilities. In this talk, I will present insights into controls on processes of
microbial community assembly in tailings, and compare the relative importance of
stochastic (dispersal) and deterministic (selection) factors in governing community
assembly processes to those observed in natural environments. Dispersal-based
recruitment required > 6 months to shift microbial community composition in tailings;
and in the absence of targeted inoculants, recruitment was dominated by species
tolerant of high salinity and extreme pH. Cell counts also remained very low ( 6 months’ equivalent) dust addition rates were required to effect growth, or
even stabilisation, of microbial cell counts in tailings. Our work identified that both
dispersal limitation and strong selection pressures play significant roles in delaying
microbial colonization and community succession in tailings.
9
The gut microbiome of an ageing-host: friend or foe?
Parag Kundu1,2
1Singapore Centre on Environmental Life Sciences Engineering, Singapore
2Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore
The gut microbiota evolves as the host ages, yet the effects of these microbial
changes on host physiology and energy homeostasis are poorly understood. To
investigate the effects, we transplanted microbiota from old or young donor mice into
young germ-free recipients. Both groups showed similar weight gain and skeletal
muscle mass, but germ-free mice transplanted with old microbiota unexpectedly
showed increased adult neurogenesis and intestinal growth. Metagenomic analysis
revealed significant enrichment in butyrate-producing microbes in the old microbiota
transplanted mice. The higher concentration of microbiota-derived butyrate in the old
microbiota transplanted mice was associated with elevated levels of the pro-
longevity hormone FGF21, which correlated with hepatic AMPK and SIRT-1
activation and with reduced mTOR signalling in these mice. Our results reveal
signatures embedded in the gut microbiome that can support the eukaryotic host
during ageing.
10The microbiome of microbialite-forming microbial mats at Rottnest Island Juliana Mendes-Monteiro1, Ryan Vogwill2, Emiley Eloe-Fadrosh3, Shane Kearney4, Cassyana Gray4, Deirdre Gleeson1 1School of Agriculture and Environment, The University of Western Australia, Crawley WA, Australia 2Hydrogeoenviro Pty, Perth, Australia 3DOE Joint Genome Institute, Walnut Creek, California, USA 4Rottnest Island Authority, Fremantle WA, Australia Microbialites are carbonate deposits that form through interactions between microorganisms and their local environment. They are classified based on their internal fabric and are generally described as being laminated stromatolitic or clotted thrombolitic structures. Microbialite forming microbial mat communities are the basis for these structures where several key microbial metabolisms influence the net carbonate precipitation and dissolution in the microbialites. Although living microbialites are relatively rare, Western Australia has one of the largest collections of microbialites which includes the stromatolites at Shark Bay, the thrombolites within the Yalgorup lakes south of Perth and the microbialites present in the salt lakes of Rottnest Island. The microbialites of Rottnest Island are considered to be a Priority Ecological Community and are potentially under threat due to anthropogenic activities on the Island, in particular increased nutrient inputs into the lakes. This work aimed to characterise the existing microbiome of microbialite-forming microbial mats across a number of lakes on Rottnest Island to serve as a base-line for assessing future changes due to anthropogenic activities. DNA was extracted from 11 separate microbial mats across 5 different lakes and sequenced on an Illumina HiSeq2500; metagenomic sequences were filtered and trimmed prior to assembly. Assembled sequences were then annotated using the Metagenomics Rapid Annotation (MG-RAST) pipeline. Multivariate analysis of sequence data revealed taxonomic and functional dissimilarities among microbial mats with significant differences (P
Profiling the diversity of Cryptosporidium species and
genotypes in wastewater treatment plants in Australia
using next generation sequencing
Alireza Zahedi1, Alexander Gofton1, Telleasha Greay1, Paul Monis2, Charlotte
Oskam1, Andrew Ball3, Andrew Bath4, Andrew Watkinson5,7, Ian Robertson1,6,
Una Ryan1
1School of Veterinary and Life Sciences, Murdoch University, Perth, Australia
2AustralianWater Quality Centre, South Australian Water Corporation, Adelaide, Australia
3WaterNSW, Sydney, Australia
4Water Corporation, Perth, Australia
5Seqwater, Ipswich, Queensland, Australia
6China-Australia Joint Research and Training Centre for Veterinary Epidemiology, College of Veterinary
Medicine, Huazhong Agricultural University, Wuhan, China
7
University of Queensland, St Lucia, Queensland, Australia
Wastewater recycling is an increasingly popular option in worldwide to reduce
pressure on water supplies due to population growth and climate change.
Cryptosporidium spp. are among the most common parasites found in wastewater
and understanding the prevalence of human-infectious species is essential for
accurate quantitative microbial risk assessment (QMRA) and cost-effective
management of wastewater. The present study conducted next generation
sequencing (NGS) to determine the prevalence and diversity of Cryptosporidium
species in 730 raw influent samples from 25 Australian wastewater treatment plants
(WWTPs) across three states: New South Wales (NSW), Queensland (QLD) and
Western Australia (WA), between 2014 to 2015. All samples were screened for the
presence of Cryptosporidium at the 18S rRNA (18S) locus using quantitative PCR
(qPCR), oocyst numbers were determined directly from the qPCR data using DNA
standards calibrated by droplet digital PCR, and positives were characterized using
NGS of 18S amplicons. Positives were also screened using C. parvum and C.
hominis specific qPCRs. The overall Cryptosporidium prevalence was 11.4%
(83/730): 14.3% (3/21) in NSW; 10.8% (51/470) in QLD; and 12.1% (29/239) in WA.
A total of 17 Cryptosporidium species and six genotypes were detected by NGS. In
NSW, C. hominis and Cryptosporidium rat genotype III were the most prevalent
species (9.5% each). In QLD, C. galli, C. muris and C. parvum were the three most
prevalent species (7.7%, 5.7%, and 4.5%, respectively), while in WA, C. meleagridis
was the most prevalent species (6.3%). The oocyst load/litre ranged from 70 to
18,055 oocysts/L (overall mean of 3,426 oocysts/L: 4,746 oocysts/L in NSW; 3,578
oocysts/L in QLD; and 3,292 oocysts/L in WA). NGS-based profiling demonstrated
that Cryptosporidium is prevalent in the raw influent across Australia and revealed a
large diversity of Cryptosporidium species and genotypes, which indicates the
potential contribution of livestock, wildlife and birds to wastewater contamination.
12Seeding of the fetal microbiome
Lisa Stinson1, Matthew Payne1, Jeffrey Keelan1
1Division of Obstetrics and Gynaecology, The University of Western Australia, Perth, WA, Australia
Background: Early-life microbial colonisation is believed to play a role in immune
programming and later life health, yet the evidence regarding the origins, timing and
significance of the neonatal microbiome remains inconclusive due to problematic
study design, biased amplification and contamination.
Method: Placental, amniotic fluid, first-pass meconium and cord blood samples were
collected from 50 elective Caesarean section deliveries. An optimised sampling and
analysis protocol was employed to minimise/control for contamination, allowing
characterisation of the fetal gut microbiome and its relationship with maternal health
parameters, short chain fatty acid (SCFA) levels and fetal immune responses.
Results: All meconium samples contained detectable levels of bacterial DNA and
the immunomodulatory SCFAs acetate and propionate, confirming the hypothesis
that the fetal gut is inoculated with bacteria/bacterial DNA in utero. At the phylum
level, meconium was dominated by Proteobacteria and Firmicutes. Pelomonas -
known to be protective against pathogenic fungal infections - was the most abundant
genus. Importantly, this genus has been found in the core non-pregnant endometrial
microbiome, but not paired vaginal samples. Lactobacillus (which dominates the
vaginal microbiome) was found in only 5 samples. Maternal atopic disease was
associated with decreased staphylococcus abundance in the fetal gut. The amniotic
fluid microbiome was distinct from the meconium microbiome and was dominated by
skin commensals, suggesting early niche construction.
Conclusions: Seeding of the fetal microbiome commences prenatally and may
originate from the endometrial microbiome present at time of conception; vaginal
contribution appears minimal. Maternal health may influence fetal immune
programming via modulation of the fetal microbiome and immunomodulatory SCFAs.
Microbial niche differentiation likely begins prior to birth.
13Long term effects of breastfeeding on immune health by
the shaping of gut microbiota
Valerie Verhasselt1
1School of Molecular Sciences, The University of Western Australia
Gut immune function conditions development of diseases that result from defects in
immune regulation such as allergic and obesity-related disease. As epidemiological
studies support the developmental origin of health and disease, the deciphering of
the critical factors modulating gut immune development should allow the advance of
primary prevention strategies specifically adapted to early life immune system. Here,
I will present gut mucosal immunity development and cover in more details the
recent understanding of the impact of early nutrition on this process. I will emphasize
how nutrition, and in particular breastmilk, can shape microbiota composition and
metabolic function and thereby their production of metabolites with immune-
modulatory properties. I will conclude with recent data from our team showing that
colostrum may be determinant in gut microbiota shaping and thereby long term
health.
14Seasonal dynamics of eukaryotic and prokaryotic diversity
in an acid saline lake
Noor-ul-Huda Ghori1, Kang Tam1, Andrew Whiteley1, Michael Wise2
1School of Agriculture and Environment (SAgE), The University of Western Australia
2School of School of Physics, Mathematics & Computing, The University of Western Australia
Lake Magic, located on the Yilgarn Craton in southern Western Australia, is a round
lake with ~1km diameter and harbours one of the most unique environments on
earth. It exhibits co-stressors of pH 1.6-4.5 and salinity of 32% TDS. The lake is
known to have the highest concentration of dissolved aluminium, iron and silica in
the world. The hydrology of Lake Magic is dynamic and undergoes stages of
flooding, evapo-concentration and desiccation, depending on local seasons. Thus, a
large population of halophilic, halotolerant and acidophilic organisms interacting to
drive key biogeochemical cycles are expected to reside in the lake, but even
rudimentary classification of the diversity present is lacking. Moreover, the dynamics
of microbial diversity during various lake stages is unclear. Here, we used a temporal
approach to understand prokaryotic and eukaryotic diversity in lake sediment and
salt mat via high-throughput amplicon sequencing of 16S rRNA and 18S rRNA gene.
The results indicated that the microbial diversity and composition are significantly
affected by lake conditions. We also observed that a vast majority of eukaryotic
diversity remains uncharacterized. Furthermore, it was seen through the different
time points that the sediment and salt mat communities evolved, becoming more
specialised in buffering the increased acidity in the lake, as a strategy to survive.
The results of this study provide preliminary understanding of the survival strategies
found in the acidic, hypersaline lake microbiome. Studying these communities
enables us to understand organisms living at the limits of life.
15Early cystic fibrosis lung disease: effects of the early
microenvironment on bacterial populations
Stephen Stick1
1Telethon Kids Institute Respiratory Research Centre
Destructive airways disease presents in children with cystic fibrosis (CF) early in life.
Bronchiectasis, the main cause of morbidity and mortality is a progressive
consequence of inflammation and infection. Little is known about how destructive
chronic infection with pathogens develops. The Australian Respiratory Early
Surveillance Team for Cystic Fibrosis (AREST CF), early surveillance program, has
provided unique insights into the relations between the early airway
microenvironment, inflammation and infection.
Early disease is characterised by an increased mucus burden, including “permanent”
mucus flakes, and inflammatory and hypoxic markers without conventional CF
bacterial pathogen infection or structural lung disease. The AREST CF data suggest
a progression of the lower airways microbiome with age, beginning with relatively
sterile airways in infancy. By age two, bacterial sequences associated with the oral
cavity dominate lower airways samples in many subjects. The presence of an oral-
like lower airways microbiome correlates with a significant increase in bacterial
density and inflammation. These early changes can occur despite the use of
antibiotic prophylaxis during the first two years of life. The majority of CF subjects
older than four harbour a pathogen dominated airway microbiome, associated with
increased inflammation and the onset of structural lung disease, despite a negligible
increase in bacterial density. The most likely explanation for the transition is a
process of microaspiration and further adaption of the microbiome to the local airway
environment. One important aspect is the effect of aspirated bile salts on the
emergence of chronic, biofilm forming organisms.
16The benefits and pitfalls of using amplicon next-generation
sequencing to investigate tick microbiomes
Telleasha Greay1, Una Ryan1, Charlotte Oskam1, Peter Irwin1
1Vector and Waterborne Pathogens Research Group, School of Veterinary and Life Sciences, Murdoch
University, Perth, WA, Australia
The tick microbiome comprises communities of microorganisms, including viruses,
bacteria and eukaryotes, and is being elucidated through modern molecular
techniques. The advent of next-generation sequencing (NGS) technologies has
enabled the genes and genomes within these microbial communities to be explored
in a rapid and cost-effective manner. The advantages of using NGS to investigate
microbiomes surpass the traditional nonmolecular methods that are limited in their
sensitivity, and conventional molecular approaches that are limited in their scalability.
Amplicon NGS has been used to study tick microbiomes over the last seven years,
and compared to shotgun NGS, remains the most popular tool to study tick
microbiomes. The advantages and disadvantages of amplicon NGS strategies that
are used to investigate tick microbiomes will be discussed during this presentation,
with a focus on bacterial and eukaryotic community characterisation. The choice of
NGS platforms, selection of library preparation techniques, the hypervariable regions
targeted, and the bioinformatic pipelines applied will be discussed, and how each of
these variables can influence estimates of the composition and diversity of microbial
communities will be considered. Recent discoveries of bacteria that are pathogenic,
symbiotic, novel and associated with tick internal organs will be highlighted. Aspects
of the tick microbiome that are largely unexplored and factors that influence the
structure and organisation of the tick microbiome will also be discussed.
17Using ‘omics technologies to understand the pathogenesis
and seek alternative therapies for otitis media in children
Rachael Lappan1, 2, Sarra Jamieson3, Christopher Peacock1,2
1The Marshall Centre for Infectious Diseases Research and Training, School of Biomedical Sciences, The
University of Western Australia, Perth, WA, Australia
2Wesfarmers Centre of Vaccines and Infectious Diseases, Telethon Kids Institute, The University of W estern
Australia, Perth, WA, Australia
Background: Recurrent acute otitis media (rAOM, recurrent ear infection) is a
common childhood disease caused by bacteria known as otopathogens, for which
current treatments have limited effectiveness. Generic probiotic therapies for rAOM
have shown promise, but lack specificity. Children with rAOM may be colonised with
one or more of the three major otopathogens (Moraxella catarrhalis, Streptococcus
pneumoniae and non-typeable Haemophilus influenzae) however some children with
rAOM do not appear to carry any of these organisms. We hypothesised that 1) novel
otopathogens may be found in the rAOM-prone middle ear and 2) healthy children
carry protective commensal bacteria in their nasopharynx, which could be useful in
the development of specific probiotic therapies.
Methods and results: We explored the microbiome of the middle ear in children
with rAOM and of the nasopharynx in children with and without rAOM using 16S
rRNA gene profiling and shotgun metagenomics. Alloiococcus otitidis and Turicella
otitidis were the dominant organisms in the middle ear, but were also abundant in the
ear canal. Corynebacterium pseudodiphtheriticum and Dolosigranulum pigrum co-
occurred abundantly in the nasopharynx of healthy children compared to children
with rAOM. C. pseudodiphtheriticum was capable of inhibiting M. catarrhalis strains
in vitro, but did not appear to produce bacteriocin-like substances.
Conclusions: C. pseudodiphtheriticum and D. pigrum are characteristic of a healthy
nasopharyngeal microbiome, and may be involved in preventing the development of
rAOM. A. otitidis and T. otitidis are possibly novel otopathogens, though their
prevalence in the ear canal means that their role as normal aural flora cannot be
ruled out.
18See the microbial world in high resolution
James Miller1, Cheryl Heiner2, Helle Christophersen1
1Millennium Science, Mulgrave, VIC, Australia
2PacBio, Menlo Park, California, USA
The ability to identify and understand the functions of the complex microbial
populations living in, on, and around us requires comprehensive characterization of
each community member. Long reads, high accuracy, and single-molecule resolution
make Single Molecule, Real-Time (SMRT) Sequencing ideal for full-length 16S rRNA
sequencing, long-read metagenomic profiling, and shotgun metagenomic assembly.
Single Molecule, Real-Time (SMRT) Sequencing for metagenomics pairs long reads
with high accuracy to:
Understand key community functions by sequencing complete operons at
99.9% accuracy with long-insert metagenomic profiling – no assembly
required
Identify microbial community members with strain-level resolution using full-
length 16S rDNA or shotgun sequencing
Discover novel genes and gene clusters by reconstructing multi-kilobase long
contigs with whole genome shotgun metagenomic assembly
Leverage epigenomic data to associate contigs and plasmids from closely
related strains
Examples of the use of SMRT sequencing to achieve the above aims will be
presented.
19Long-term Paleolithic diet is associated with low resistant starch, high saturated fat intake, increased TMAO concentrations and different gut microbiota composition Angela Genoni1 1School of Medical and Health Sciences, Edith Cowan University, Joondalup WA, Australia The Paleolithic diet (PD) is promoted for improved gut health. Although, it excludes grains and dairy, groups that form part of the AGHE. To evaluate this in a cross- sectional study we recruited n=44 long-term (>1year) PD followers, and 44 controls, who followed an Australian diet. Three-day weighed diary records, blood and stool samples were collected to evaluate if long-term PD altered gut markers and the microbiota, serum trimethylamine-N-oxide (TMAO) and how that related to dietary intake. There was variability in adherence to the Paleolithic diet, and the PD group was stratified into Strict Paleolithic (SP) (n=22) and Pseudo-Paleolithic (PP) (n=22) groups. RS intake was lower and saturated fat intake higher in both Paleolithic groups, compared to controls (P
Echoes of the microbiome: irritable bowel syndrome
diagnosis through bowel sounds
Mary Webberley1, Xuhao Du1, Gary Allwood1, Andrisha-Jade Inderjeeth2, Adam
Osseiran3, Barry Marshall1,2
1The Marshall Centre, The University of Western Australia, WA
2SirCharles Gairdner Hospital, Nedlands, WA
3School of Engineering, Edith Cowan University, WA
Irritable bowel syndrome is a common functional gut disorder causing abdominal
pain and altered bowel habits. IBS aetiology is multi-factorial, but in around 50% of
cases it appears to originate in the gut. Changes in the gut microbiota are associated
with alterations in gut permeability, immunomodulation, food processing and nervous
system changes affecting motility and visceral perception that ultimately result in IBS
symptoms. These changes also affect the sounds produced by the gut.
Typically, IBS is a diagnosis of exclusion after patients undergo colonoscopy to
exclude organic disease. Clinicians have identified a need for a new cost-effective,
accurate and non-invasive diagnostic test for IBS.
We explored the use of bowel sounds to characterize IBS with a view to diagnostic
use. We recruited participants with an existing clinical diagnosis of IBS, or healthy
(asymptomatic) digestive systems and recorded their bowel sounds for two hours
and 40 minutes.
We report our results including our accuracy in characterizing IBS related bowel
sounds and differentiation between IBS patients and healthy participants. Leave-
one-out cross-validation of our model developed using the first 68 participants gave
90% sensitivity and 92% specificity for IBS diagnosis. Independent testing using the
next 15 IBS and 15 healthy participants demonstrated 87% sensitivity and 87%
specificity for IBS diagnosis.
These preliminary results provide proof-of-concept for the use of bowel sounds
analysis for the diagnosis of IBS. Our belt and model offer hope of a new, more
accurate alternative for positive and non-invasive diagnosis of IBS.
21Sampling scale, scope and resolution affect interpretation
of termite core microbiomes
Ghislaine Platell1, Katharine Howell1, Boris Baer2, Tamara Hartke3
1AustralianResearch Council Centre of Excellence in Plant Energy Biology, The University of Western Australia
2Centrefor Integrative Bee Research (CIBER), Department of Entomology, University of California
3Blumenbach Institute for Zoology and Anthropology, University of Göttingen
Core microbiomes, the conserved gut community thought to be critical to colony
function, have previously been reported for various termites. A number of different
methods have been used, however, which may affect interpretation of results across
studies. We tested how the number and types of samples affect core community
estimation using two local higher termite species: Tumulitermes westraliensis and
Amitermes obeuntis. Overall community richness in A. obeuntis was around 1.5
times that of T. westraliensis. In both species, the number of bacterial OTUs in the
core community and the abundance of these taxa was inversely related to the
number, caste composition, and spatial distribution of samples used in the
determination. Single-caste core communities were larger than all-caste core
communities and always included taxa unique to that caste. However, the number of
samples included was found to have a greater effect on core community size than
sample type. These results underline the importance of ensuring that sampling scale,
scope and resolution for termite gut microbiome studies is appropriate to the
question under investigation. We recommend a minimum of twenty replicates to
estimate the core community more accurately and the types of samples to be
included depend on the ecological question.
22Life after death: thanatomicrobiome and its application in
forensic science
Eric Benbow1, Paola Magni2
1Department of Entomology, Department of Osteopathic Medical Specialties, Michigan State University, East
Lansing, MI-USA
2School of Veterinary & Life Sciences, Murdoch University, WA
Human antemortem microbiotas are well documented by the Human Microbiome
Project (Peterson et al., 2009), for various body locations of healthy individuals. But
what happen to the microbiota when the body dies? While death is considered a
universal phenomenon, when it comes to thanatomicrobiome (thanatos-, Greek for
death) and epinecrotic communities (microbial communities residing in and/or
moving on the surface of decomposing remains), “life after death” may be
considered a reality. Over the last few years microbiologists are developing new
applications to investigate the dynamic and coordinated changes in microbial activity
that occur when a human host dies. Such results may be applicable in forensic
investigations, in particular regarding the estimation of the time since death (PMI,
Post Mortem Interval), a pivotal information for the crime scene reconstruction. This
presentation aims to provide a review of emerging state of the art research relating
to thanatomicrobiome and epinecrotic communities and discuss the possibility to
develop similar research in WA.
23Isoquercetin and inulin synergistically modulate the gut
microbiome to prevent development of the metabolic
syndrome in mice fed a high fat diet
Si Tan1,2, Jose Caparros-Martin3, Vance Matthews2, Henrietta Koch2, Fergal
O’Gara3,4, Kevin Croft2, Natalie Ward3,5
1LifeScience and Technology Institute, Yangtze Normal University, Chongqing China
2School of Biomedical Sciences, The University of Western Australia, Perth, Australia
3School of Biomedical Sciences & Curtin Health Innovation Research Institute, Curtin University, Perth, Australia
4Biomerit Research Centre, School of Microbiology, National University of Ireland, Cork Ireland
5Medical School, The University of Western Australia, Perth, Australia
Aims/hypothesis: Dietary fibre positively influences gut microbiome composition,
enhancing the metabolism of dietary flavonoids to produce bioactive metabolites.
These synergistic activites facilitate the beneficial effects of dietary flavonoids on
cardiometabolic health parameters. The aims of this study were to investigate
whether isoquercetin (a major dietary flavonoid) and inulin (soluble fibre), either
alone or in combination could improve features of the metabolic syndrome.
Methods: Following a 1 week acclimatization, male C57BL6 mice (6-8 weeks) were
randomly assigned to; (i) normal chow diet (n=10), (ii) high fat (HF) diet (n=10), (iii)
HF diet + 0.05% isoquercetin (n=10), (iv) HF diet + 5% inulin, or (v) HF diet + 0.05%
isoquercetin + 5% inulin (n=10). Body weight and food intake were measured
weekly. At 12 weeks, glucose and insulin tolerance tests were performed, blood,
faecal samples, liver, skeletal muscle and adipose tissue were collected.
Results: At 12 weeks, mice on the HF diet had significantly elevated body weights
as well as impaired glucose tolerance and insulin sensitivity compared to the normal
chow mice. Supplementation with either isoquercetin or inulin had no effect, however
mice receiving the combination had attenuated weight gain, improved glucose
tolerance and insulin sensitivity, reduced hepatic lipid accumulation, adipocyte
hypertrophy, circulating leptin and adipose FGF21 levels, comparable to mice
receiving the normal chow. Additionally, mice on the combination diet had
improvements in the composition and functionality of their gut microbiome as well as
production of short chain fatty acids.
Conclusions/interpretation: Long-term supplementation with the dietary flavonoid
isoquercetin and the soluble fibre inulin can prevent development of the metabolic
syndrome in mice fed a high fat diet. This protective effect appears to be mediated,
in part, through beneficial changes to the microbiome.
24Quality assurance in P4 systems medicine: the impact of
preanalytical processes, biobanking protocols and
analytical platform stability
David Broadhurst1
1Centre for Integrative Metabolomics & Computational Biology, Edith Cowan University, WA
The post-genomic technologies underpinning P4 Systems Medicine can be affected
by long-term platform stability, together with methods of sample collection, storage,
and handling. Thus, it is vitally important to consider the requirements for
preanalytical processes and biobanking before embarking on any research project in
this area. Poor practice can create unnecessary confounding factors and excessive
measurement error, with deleterious effects on the robustness and reproducibility of
acquired data. Here I present the current best practice and latest evidence on
preanalytical processes and biobanking of samples intended for
metabolomics/microbiome measurement of common biofluids and tissues. It
highlights areas requiring more validation and research and provides some
evidence-based guidelines on best practices.
25Flash talks
Phylogenetic and predicted functional profile of microbial
communities in biological soil crusts of the Midwest in
Western Australia
Kang Tam1,2, Benjamin Moreira-Grez2, Wei San Wong1,2, Adam Cross1, Jean
Yong1,2, Andy Whiteley2
1Centre for Mine Site Restoration, Curtin University, Bentley WA
2Faculty of Science, The University of Western Australia, Crawley WA
Biological soil crusts (BSCs) are communities of cyanobacteria, lichen, algae, fungi,
and bryophyte commonly found in arid and semiarid regions. It was estimated that
they cover approximately 12% of global terrestrial surfaces (Rodriguez-Caballero et
al., 2018). Recently, BSCs have received global attention due to their diverse roles
as ecosystem engineers. Functionally, they stabilise soil, reduce erosion, regulate
topsoil hydrology, fix atmospheric carbon and nitrogen, and in some situations,
promote vascular plant growth. The advent of next-generation sequencing has
provided exciting opportunities to characterise the microbial constituents of BSCs
and their associated functional genes. Despite two-thirds of Australia’s land mass
being comprised of arid and semiarid climate, relatively little is known about native
BSCs in Western Australia. In this study, we collected BSCs and bare soil from the
Midwest region of Western Australia. Bacterial communities were characterised
through ion torrent sequencing of 16S rRNA amplicons. Furthermore, we inferred
functional genes present within each BSC type using the bioinformatics package,
PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of
Unobserved States). Following that, the phylogenetic and functional profiles of BSCs
and bare soil were compared.
26The impact of colostrum on growth
Lieke van den Elsen1, Akila Rekima1, Charlotte Isnard2, Valerie Verhasselt1
1School of Molecular Sciences, The University of Western Australia, Perth, WA
2University of Nice, Nice, France
Stunting affects almost 25% of children under 5 years of age, and is associated with
lifelong consequences such as poor cognition and chronic diseases. This condition is
driven by multiple factors including undernutrition and repeated infections.
Colostrum, the very first fluid produced by the mammary gland, is very rich in anti-
infectious molecules and growth factors compared to more mature milk. However,
colostrum-feeding practises are sub-optimal worldwide. We propose that delayed
initiation of breastfeeding, and hence lack of colostrum feeding, contributes to
stunting. We compared growth of mice fed physiologically from birth (control group)
with mice fostered to a dam that delivered 8 days earlier (colostrum minus (colminus))
and thus received mature milk only. Colminus mice had lower body weight gain
compared to control mice from as early as 3 days old. The weight difference peaked
around day 15, with 20% deviation from the control group, and normalized into
adulthood. To assess if the difference in growth was driven by alterations in neonatal
gut microbiota, fecal samples from preweaned mice were used for 16S rDNA
bacterial sequencing. Colminus mice showed marked alterations in gut microbiota
composition compared to control mice, including a pronounced reduction in
Bacteroides acidifaciens (BA) abundance. Further experiments will determine
whether immune dysfunction due to lack of BA is responsible for altered metabolism
in mice deprived of colostrum. Ultimately, this work should provide the basis to
promote colostrum feeding as an effective and inexpensive way to support healthy
growth in developing countries.
27Targeted amplicon next generation sequencing for
screening and genotyping of pathogens in cattle
Megan Jordan1, Sam Hair1, Mengqi Chen1, Shane Besier1, Michael Bunce2,
David Williams3, Nicky Buller1
1Departmentof Primary Industries and Regional Development, South Perth, WA
2TrEnDLaboratory, School of Molecular and Life Sciences, Curtin University, Bentley, WA
3CSIRO Australian Animal Health Laboratory, Geelong, VIC
Accurate and rapid diagnostic methods are vital for determining herd disease
prevalence and for the surveillance and control of infectious diseases to mitigate
biosecurity risks. Currently, multiple diagnostic panels and sample submissions are
required to determine the disease status of a herd; this is both costly and time
consuming. A Targeted Amplicon Sequencing (TAS) approach, utilising Next
Generation Sequencing technology, can simultaneously test for multiple disease
agents, offering an efficient and cost effective alternative to undertaking a plethora of
individual diagnostic assays. In addition, the TAS method can detect and
differentiate host specific pathogen strains, which is important in disease
management. TAS also overcomes some of the challenges of a whole genome
sequencing approach to diagnostics, which requires deep sequencing, significant
computational power and the storage of very large data sets. We developed a TAS
protocol for the detection of 20 different bacterial, viral and parasitic pathogens
causing respiratory, intestinal and reproductive diseases in cattle. Results of the TAS
method were compared with those of current standard diagnostic tests including
culture, PCR and ELISA. Sensitivity was compared with qPCR in four representative
pathogens. Results showed TAS to be a sensitive, effective and rapid method that
was capable of detecting multiple pathogens in clinical samples with mixed infections
and strain typing (where required) using the sequence information generated. The
results demonstrate the feasibility of the method and show it is a useful approach for
veterinary disease diagnostics and biosecurity management.
28Characterisation of the human milk microbiome from
mothers who delivered preterm and term infants
Ali Sadiq Cheema1, Megan Lloyd2,5, Richard Allcock3, Stephanie Trend4, Chooi
Heen Kok4, Karen Simmer4,5, Matthew Payne5, Donna Geddes1
1School of Molecular Sciences, The University of Western Australia
2School of Medical and Health Sciences, Edith Cowan University
3School of Biomedical Sciences, The University of Western Australia
4Centre for Neonatal Research and Education, The University of Western Australia
5Medical School, The University of Western Australia
Introduction: Human milk (HM) is a source of commensal bacteria that may
colonize the infant gut and enhance growth and development of infants.
Objectives: To characterize the bacterial diversity in HM collected in the first month
postpartum from mothers who delivered preterm and term infants.
Procedures: DNA in the milk samples from mothers who delivered preterm (n=58,
37 weeks) infants was extracted using the QIAamp
DNA Mini Kit and quantified on a Qubit fluorometer. The V4 region of 16S rRNA
gene was amplified and sequenced on an Ion Torrent PGM. Reads were analysed
using the Greenfield Hybrid Analysis Pipeline v2.1, which incorporates USEARCH
and the Ribosomal Database Project sequence database. Additional statistical
analyses were performed with Microbiome Analyst.
Results: In preterm samples, the most abundant phyla were the Firmicutes (42.3%),
Proteobacteria (34.2%), Actinobacteria (1.2%) and Bacteroidetes (1.0%) while in
term samples these were Proteobacteria (44.7%), Firmicutes (39.2%), Actinobacteria
(1.0%) and Bacteroidetes (2.2%). In preterm samples, the most abundant genera
were Staphylococcus sp. (39.0%), Acinetobacter sp. (17.2%), Pseudomonas sp.
(13.1%), Streptococcus sp. (1.7%) and Aeromonas sp. (0.01%), while in term
samples, they were Staphylococcus sp. (18.4%), Acinetobacter sp. (17.3%),
Streptococcus sp. (16.9%), Pseudomonas sp. (12.7%) and Aeromonas sp. (5.3%).
Principal coordinate analysis (weighted unifrac distance) showed maternal BMI,
mode of delivery and gestation age does not affect bacterial diversity in either
preterm or term HM.
Conclusions: This sample set does not show differences in the bacterial diversity of
preterm and term HM. Further factors such as maternal BMI, mode of delivery and
gestational age did not influence bacterial diversity.
29Poster presentations [P1] What’s in a tick? Meta-analysis of microbial diversity in Australian ticks Siobhon Egan1, Telleasha Greay1, Alexander Gofton1, Una Ryan1, Peter Irwin1, Charlotte Oskam1 1Vector and Waterborne Pathogens Research Group, School of Veterinary and Life Sciences, Murdoch University, Perth, Australia Ticks (Acari: Ixodida) are a group of blood-feeding arthropods and approximately 10% of tick species transmit pathogens, affecting livestock, domestic animals and humans worldwide. The majority of tick-borne pathogens (TBPs) are considered zoonotic, meaning that pathogens within ticks can be transmitted between animals and humans, causing disease. Australia has experienced growing concern about the presence of an unknown tick-borne disease in humans. Molecular advances, such as next generation sequencing (NGS), have recently identified a number of candidate TBPs in Australian ticks. Through a rigorous bioinformatics analysis, this project aimed to determine what factors influence the bacterial communities of Australian ticks and evaluate patterns of candidate TBPs. Amplicon NGS targeting the ubiquitous bacterial 16S rRNA gene was performed on the Illumina MiSeq platform (n = 1208). Analysis revealed over 64 million assigned sequences and showed that tick species was the most significant factor (P
[P2] Identification of a novel conserved quorum-sensing
locus producing unsaturated N-acyl-homoserine lactones
Tahlia Bastholm1, Nigel Halliday2, Alex Truman2, Louise Thorne3, Timothy
Haskett4, Jason Terpolilli4, Clive Ronson3, Paul Williams2, Joshua Ramsay1
1Curtin Health Innovation Research Institute and School of Pharmacy and Biomedical Sciences, Curtin
University, Perth, WA, Australia
2School of Life Sciences and Centre for Biomolecular Sciences, University of Nottingham, Nottingham, United
Kingdom
3Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand
4Centre for Rhizobium Studies and School of Veterinary and Life Sciences, Murdoch University, Perth, WA,
Australia
Bacteria use intercellular chemical communication systems called quorum sensing
(QS), to control phenotypes such as biofilm formation and virulence at multicellular
level. Membrane-diffusible N-acyl-homoserine lactones (AHLs) produced by bacteria
increase in concentration with increasing cell density. AHLs bind cytoplasmic
receptor proteins, which activate phenotypes under QS control. We have
characterised a novel QS locus in the nitrogen-fixing Mesorhizobium spp. named
mqsRIC, which appears completely conserved across this genus. Uniquely, mqsC
encodes an AHL modification enzyme MqsC, which together with AHL synthase
MqsI, produces an AHL to activate the receptor MqsR. MqsC - a crotonase-family
enzyme - resembles proteins responsible for synthesis of the distinct ‘diffusible
signal factor’ (DSF) family of QS systems found in plant and animal pathogens, such
as Xanthomonas and in Burkolderia spp., respectively. DSF synthases introduce
double bonds into fatty acids to produce cis-2-unsaturated fatty acids. Here we
confirm that the MqsR receptor is activated by the unsaturated AHL, 5,6-cis-
dodecenoyl-homoserine-lactone (5-cis-C12-HSL). Liquid chromatography coupled to
tandem mass spectrometry (LC-MS/MS) revealed M. loti supernatants contained a
molecule with the same mass as chemically synthesised 5-cis-C12-HSL.
Synthesised 5-cis-C12-HSL added to bioassays activated MqsR in an mqsC-deletion
mutant, however, C12-HSL and other analogues did not. Interestingly, C12-HSL did
weakly activate MqsR in an mqsI mutant (still carrying mqsC), suggesting MqsC
might convert the exogenously supplied C12-HSL to 5-cis-C12-HSL. In summary, we
have identified a unique QS system present throughout the mesorhizobia that may
represent an evolutionary amalgamation of synthesis pathways for two distinct QS
systems.
31[P3] Are plant growth promoting microbes compromised
by stockpiling of topsoil (in mine restoration context)?
Wei San Wong1,2, Tim Morald3, Kang Tam3, Paul Nevil2, Andy Whiteley3, Jean
Yong1,2,4
1School of Biological Sciences, The University of Western Australia, Australia
2ARC Centre for Mine Site Restoration, School of Molecular and Life Sciences, Curtin University, Australia
3School of Agriculture and Environment, The University of Western Australia, Australia
4Department of Biosystems and Technology, Swedish University of Agricultural Sciences, Sweden
Western Australian mine restoration utilises topsoil stripping prior to mining, then
stockpiling (often long-term), and subsequently re-spreading (often diluted) onto
mined-out areas. Improvements to mine restoration practices tend to examine
broadcast seed mixes or soil chemical properties (i.e. chemical fertiliser addition to
accelerate plant growth). Soil biology is often overlooked, yet soil quality and
functionality are determined by both physiochemical and biological properties.
Microbial composition and the benefits conferred to soil and plants, evident by their
positive impacts on agricultural productivity remain undervalued in mine restoration.
Beneficial soil-microbe-plant interactions to improve plant establishment and eco-
physiological resilience in mine restoration warrant further research. We hypothesize
that prolonged stockpiling of topsoil reduces microbial efficacy due to shifts in
community assemblage and decreased diversity, including potentially pivotal
symbiotic partners responsible for the production of microbial “signals” such as
phytohormones or novel metabolites. Here we examine microbial activity of
stockpiled topsoil and addition of microbe inoculum as a method of reintroducing
symbiotic partners to enhance plant (native species - Senna notabilis and Acacia
hilliana; crop species - Cajanus cajan) establishment and growth. Our study showed
mixed, nevertheless encouraging results. In order to identify community scale
microbial changes; our future experiments will seek to use Next Generation
Sequencing (NGS) to look at bacterial and fungal DNA in pre and post mining soils.
Thus, further collaborative efforts will test practical methods to introduce or
rejuvenate plant growth promoting microorganisms in stockpiled topsoil. Field
experiments trialling the efficacy of these bio-inoculums under site conditions are
also required.
32[P4] Effect of natural products on the production and
activity of Clostridium difficile toxins in vitro
Niloufar Roshan1, Thomas Riley1-4, Daniel Knight3, Katherine Hammer1
1School of Biomedical Sciences, The University of Western Australia, Crawley WA, Australia
2Department of Microbiology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Nedlands WA,
Australia
3School of Veterinary & Life Sciences, Murdoch University, Murdoch WA, Australia
4School of Medical & Health Sciences, Edith Cowan University, Joondalup WA, Australia
Backgrounds: Clostridium difficile infection is a toxin-mediated disease of the colon.
C. difficile virulence is primarily attributed to the production of toxin A and toxin B,
thus therapeutic strategies which reduce toxin production and activity can
significantly decrease the pathogenicity of infection.
Objective: To investigate the effect of a range of natural products on the production
and activity of C. difficile toxins in vitro.
Methods: Twenty-two natural products were investigated against four C. difficile
strains. The activity of products against toxins was determined using Vero and HT-29
cells cytotoxicity and neutral red uptake assays. The indirect effect of products on
toxin-mediated cytotoxicity was determined using the same cell lines. The effect of
seven products on toxin production by C. difficile was determined using ELISA.
Results: Zingerone (0.3 mg/ml) protected both cell lines from C. difficile cytopathic
effects, confirmed by the neutral red uptake assay (P < 0.05). Three Leptospermum
honeys (4% w/v), fresh onion bulb extract (12.5% v/v) and trans-cinnamaldehyde
(0.005% v/v) all reduced toxin production and activity significantly (P ≤ 0.023). Garlic
clove powder (4.7 mg/ml) only reduced toxin activity (P ≤ 0.047).
Conclusions: Five out of 22 natural products reduced both C. difficile toxin
production and activity in vitro. Zingerone showed protection against the cytopathic
effect of C. difficile toxins likely through blocking either the toxin binding sites on the
toxin molecule or the host cell receptors.
Significance and impact of the study: This study encourages further investigation
of natural products against C. difficile toxins in vivo.
33[P5] First report on whole-genome sequencing analysis, antimicrobial resistance and epidemiological features of Salmonella diarrhoeal infection among children Ali Harb1,2,3, Sam Abraham1,2, Bertha Rusdi1,2, Tanya Laird1,2, Mark O’Dea1,2, Ihab Habib1,2 1School of Veterinary and Life Sciences, Murdoch University, Perth, Australia 2AntimicrobialResistance and Infectious Diseases (AMRID) Laboratory, Murdoch University, Perth, Australia 3Thi-Qar Public Health Division, Ministry of Health, Iraq Salmonellosis is one of the most common bacterial diarrhoeal illnesses among children and poses a significant public health burden worldwide. This study investigates the occurrence, serotypes distribution, risk factors, antimicrobial resistance, and multilocus sequence types (ST) of Salmonella isolated from diarrhoea cases among children aged
[P6] Bio-cement from extremophilic microbes:
Underpinning molecular mechanisms and improving the
efficacy of self-healing concrete
Sakshi Tiwari1, Andy Whitley2, Abhijit Mukherjee1, Navdeep Dhami1
1Department of Civil Engineering, Curtin University, Bentley, Perth WA, Australia
2School of Agriculture and Environment, The University of Western Australia, Crawley, Perth WA, Australia
Microbially Induced Carbonate Precipitation (MICP) has emerged as a potential
technology for improving the durability, repair and restoration of construction
materials using bacteria. This bio-based method is a mimicry of naturally cementing
formations as corals, beach rocks, stromatolites wherein living microbes are involved
in mineralisation of carbonates at ambient temperature conditions making it a highly
sustainable cement. Most of the studies till date have been carried out using
alkalophilic cultures working optimally at pH 9 but real field conditions of concrete
reach upto 12 impeding the efficacy and viability of such cultures. In the current
study, we made an attempt to study the microbial diversity of bacteria from highly
alkaline sites as cement and calcareous soils along with an attempt to isolate
alkalophilic calcifying cultures with potential efficacy under real field conditions. We
mimicked the stress conditions of concrete to isolate extremophilic strains and
consortia by enrichment methods. The isolates were selected on the basis of their
spore formation, urease production, alkali resistance, viability and calcite
precipitation. The outcome of this work will unpin the molecular mechanisms
involved in metabolism of such cultures under alkaline environments and improve
the applications of biocement technology for concrete environments.
35[P7] High antibody titres against pneumococcal and Haemophilus influenzae vaccine candidates have no impact on nasopharyngeal carriage in Papua New Guinean children Tasmina Rahman1,2, Ruth Thornton2,3, Tilda Orami4, Peter Jacoby2, Karli Corscadden2, Rebecca Ford4, Deborah Lehmann2, William Pomat4, Peter Richmond1,2, Lea-Ann Kirkham1,2, the 10v13v PCV trial team 1Division of Paediatrics, The University of Western Australia, Perth, Australia 2Wesfarmers Centre of Vaccines and Infectious Diseases, Telethon Kids Institute, University of Western Australia, Perth, Australia 3School of Biomedical Sciences, The University of Western Australia, Perth, Australia 4Papua New Guinea Institute of Medical Research, Goroka, Papua New Guinea Papua New Guinean (PNG) children are colonised with respiratory pathogens, Streptococcus pneumoniae and Haemophilus influenzae, at an exceptionally early and high rate. This facilitates pathogen transmission and increases the risk of developing respiratory infections. Current vaccines provide limited protection against these pathogens, with ideal vaccines preventing colonisation and therefore transmission of these species. To inform vaccine development, it is important to assess the natural immune response against putative vaccine antigens in longitudinal cohorts at high risk of respiratory disease and to understand the correlation with colonisation. Sera and nasopharyngeal swabs were collected from 133 PNG children at 1, 4, 9, 10, 23 and 24 months of age. Serum IgG against pneumococcal (PspA1/2, CbpA and Ply) and H. influenzae vaccine candidates (P4, P6, OMP26 and rsPilA) was measured by multiplex bead-based immunoassay. Nasopharyngeal carriage was determined by culture. Antibody titres against all pneumococcal proteins declined between 1 month and 9 months of age (p
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