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A follow-up study was performed in 12 healthy women to evaluate systemic exposure to aluminium following topical application of a representative antiperspirant formulation under real-life use conditions (part A) and to assess the local fate of topically applied aluminium by taking additional tape strips and skin biopsies (Part B). A simple roll-on formulation, containing the maximal possible radioactive dose, was prepared with [26Al] aluminium-labeled chlorohydrate (ACH). The microtracer of [26Al] was used to distinguish aluminium from the natural background, using accelerator mass spectrometry. [26Al] aluminiumcitrate was administered intravenously to estimate the dermal fraction absorbed. Despite the 25-fold increase of the topical dose compared with the previous study, only 12 blood samples gave results above the lower limit of quantitation (0.118 fg/mL). The most reliable estimates of the dermal fraction absorbed are derived from noncompartmental analysis with the urine data. By using the intravenous dose to normalize the urinary excretion to 100% bioavailability, the best estimate of the fraction absorbed of [26Al] from a topical application of [26Al]-aluminium-labeled chlorohydrate in an antiperspirant formulation was 0.00052%. Part B of the study demonstrated that the majority of the aluminium in the formulation remained associated with the external layers of the skin without penetration through the skin.

Background A novel human parechovirus 3 Australian recombinant (HPeV3-AR) strain emerged in 2013 and coincided with biennial outbreaks of sepsis-like illnesses in infants. We evaluated the molecular evolution of the HPeV3-AR strain and its association with severe HPeV infections. Methods HPeV3-positive samples collected from hospitalized infants aged 5–252 days in 2 Australian states (2013–2020) and from a community-based birth cohort (2010–2014) were sequenced. Coding regions were used to conduct phylogenetic and evolutionary analyses. A recombinant-specific polymerase chain reaction was designed and utilized to screen all clinical and community HPeV3-positive samples. Results Complete coding regions of 54 cases were obtained, which showed the HPeV3-AR strain progressively evolving, particularly in the 3′ end of the nonstructural genes. The HPeV3-AR strain was not detected in the community birth cohort until the initial outbreak in late 2013. High-throughput screening showed that most (>75%) hospitalized HPeV3 cases involved the AR strain in the first 3 clinical outbreaks, with declining prevalence in the 2019–2020 season. The AR strain was not statistically associated with increased clinical severity among hospitalized infants. Conclusions HPeV3-AR was the dominant strain during the study period. Increased hospital admissions may have been from a temporary fitness advantage and/or increased virulence.

Summary Objective To elucidate the effects of meteorological variations on the activity of influenza A and B in 11 sites across different climate regions. Methods Daily numbers of laboratory-confirmed influenza A and B cases from 2011–2015 were collected from study sites where the corresponding daily mean temperature, relative humidity, wind speed and daily precipitation amount were used for boosted regression trees analysis on the marginal associations and the interaction effects. Results Cold temperature was a major determinant that favored both influenza A and B in temperate and subtropical sites. Temperature-to-influenza A, but not influenza B, exhibited a U-shape association in subtropical and tropical sites. High relative humidity was also associated with influenza activities but was less consistent with influenza B activity. Compared with relative humidity, absolute humidity had a stronger association - it was negatively associated with influenza B activity in temperate zones, but was positively associated with both influenza A and B in subtropical and tropical zones. Conclusion The association between meteorological factors and with influenza activity is virus type specific and climate dependent. The heavy influence of temperature on influenza activity across climate zones implies that global warming is likely to have an impact on the influenza burden.

A novel human parechovirus 3 Australian recombinant (HPeV3-AR) strain emerged in 2013 and coincided with biennial outbreaks of sepsis-like illnesses in infants. We evaluated the molecular evolution of the HPeV-AR strain and its association with severe HPeV infections.HPeV3-positive samples collected from hospitalized infants aged 5-252 days in two Australian states (2013-2020) and from a community-based birth cohort (2010-2014) were sequenced. Coding regions were used to conduct phylogenetic and evolutionary analyses. A recombinant-specific PCR was designed and utilized to screen all clinical and community HPeV3-positive samples.Complete coding regions of 54 cases were obtained, which showed the HPeV3-AR strain progressively evolving, particularly in the 3' end of the non-structural genes. The HPeV3-AR strain was not detected in the community birth cohort until the initial outbreak in late 2013. High-throughput screening showed most (75%) hospitalized HPeV3 cases involved the AR strain in the first three clinical outbreaks, with declining prevalence in the 2019-20 season. The AR strain was not statistically associated with increased clinical severity amongst hospitalised infants.The HPeV3-AR was the dominant strain during the study period. Increased hospital admissions may have been from a temporary fitness advantage and/or increased virulence.

A follow-up study was performed in 12 healthy women to evaluate systemic exposure to aluminium following topical application of a representative antiperspirant formulation under real-life use conditions (part A) and to assess the local fate of topically applied aluminium by taking additional tape strips and skin biopsies (Part B). A simple roll-on formulation, containing the maximal possible radioactive dose, was prepared with [

International audience; ObjectivesTo improve our understanding of the global epidemiology of common respiratory viruses by analysing their contemporaneous incidence at multiple sites.Methods2010–2015 incidence data for influenza A (IAV), influenza B (IBV), respiratory syncytial (RSV) and parainfluenza (PIV) virus infections were collected from 18 sites (14 countries), consisting of local (n = 6), regional (n = 9) and national (n = 3) laboratories using molecular diagnostic methods. Each site submitted monthly virus incidence data, together with details of their patient populations tested and diagnostic assays used.ResultsFor the Northern Hemisphere temperate countries, the IAV, IBV and RSV incidence peaks were 2–6 months out of phase with those in the Southern Hemisphere, with IAV having a sharp out-of-phase difference at 6 months, whereas IBV and RSV showed more variable out-of-phase differences of 2–6 months. The tropical sites Singapore and Kuala Lumpur showed fluctuating incidence of these viruses throughout the year, whereas subtropical sites such as Hong Kong, Brisbane and Sydney showed distinctive biannual peaks for IAV but not for RSV and PIV.ConclusionsThere was a notable pattern of synchrony of IAV, IBV and RSV incidence peaks globally, and within countries with multiple sampling sites (Canada, UK, Australia), despite significant distances between these sites.

Meningococcal serogroup B (MenB) accounts for an important proportion of invasive meningococcal disease (IMD). The 4-component vaccine against MenB (4CMenB) is composed of factor H binding protein (fHbp), neisserial heparin-binding antigen (NHBA), Neisseria adhesin A (NadA), and outer membrane vesicles of the New Zealand strain with Porin 1.4. A meningococcal antigen typing system (MATS) and a fully genomic approach, genetic MATS (gMATS), were developed to predict coverage of MenB strains by 4CMenB. We characterized 520 MenB invasive disease isolates collected over a 5-year period (January 2007–December 2011) from all Australian states/territories by multilocus sequence typing and estimated strain coverage by 4CMenB. The clonal complexes most frequently identified were ST-41/44 CC/Lineage 3 (39.4%) and ST-32 CC/ET-5 CC (23.7%). The overall MATS predicted coverage was 74.6% (95% coverage interval: 61.1%–85.6%). The overall gMATS prediction was 81.0% (lower–upper limit: 75.0–86.9%), showing 91.5% accuracy compared with MATS. Overall, 23.7% and 13.1% (MATS) and 26.0% and 14.0% (gMATS) of isolates were covered by at least 2 and 3 vaccine antigens, respectively, with fHbp and NHBA contributing the most to coverage. When stratified by year of isolate collection, state/territory and age group, MATS and gMATS strain coverage predictions were consistent across all strata. The high coverage predicted by MATS and gMATS indicates that 4CMenB vaccination may have an impact on the burden of MenB-caused IMD in Australia. gMATS can be used in the future to monitor variations in 4CMenB strain coverage over time and geographical areas even for non-culture confirmed IMD cases.

Parabens are esters of para-hydroxybenzoic acid that have been used as preservatives in many types of products for decades including agrochemicals, pharmaceuticals, food and cosmetics. This illustrative case study with propylparaben (PP) demonstrates a 10-step read-across (RAX) framework in practice. It aims at establishing a proof-of-concept for the value added by new approach methodologies (NAMs) in read-across (RAX) for use in a next-generation risk assessment (NGRA) in order to assess consumer safety after exposure to PP-containing cosmetics. In addition to structural and physico-chemical properties, in silico information, toxicogenomics, in vitro toxicodynamic, toxicokinetic data from PBK models, and bioactivity data are used to provide evidence of the chemical and biological similarity of PP and analogues and to establish potency trends for observed effects in vitro. The chemical category under consideration is short (C1–C4) linear chain n-alkyl parabens: methylparaben, ethylparaben, propylparaben and butylparaben. The goal of this case study is to illustrate how a practical framework for RAX can be used to fill a hypothetical data gap for reproductive toxicity of the target chemical PP.

Q fever is a zoonotic disease caused by infection with Coxiella burnetii transmitted from animals including, but not limited to, cattle, sheep and goats. The infection in cattle is typically sub-clinical with some evidence suggesting associated reproductive loss. There is currently limited data on the true prevalence and distribution of coxiellosis in beef cattle across northern Australia. During this study, 2,012 sera samples from beef cattle managed on commercial farms located in Queensland and the Northern Territory were tested using an indirect immunofluorescent assay (IFA) for serological evidence of IgG antibodies against C. burnetii. Bayesian latent class models were used to estimate the true prevalence, adjusted for diagnostic test sensitivity and specificity and incorporating the hierarchical structure of the cattle within farms and regions. In this study, cattle in the Northern Territory had lower estimated true prevalence than cattle within most regions of Queensland with the exception of south-east Queensland. Results from this study have described the geographic distribution and estimated the true prevalence of antibodies to C. burnetii in a sample of extensively managed beef cattle located across the tropical grazing regions of northern Australia.

Q fever is a notifiable zoonotic disease in Australia, caused by infection with Coxiella burnetii. This study has reviewed 2,838 Q fever notifications reported in Queensland between 2003 and 2017 presenting descriptive analyses, with counts, rates, and proportions. For this study period, Queensland accounted for 43% of the Australian national Q fever notifications. Enhanced surveillance follow-up of Q fever cases through Queensland Public Health Units was implemented in 2012, which improved the data collected for occupational risk exposures and animal contacts. For 2013–2017, forty-nine percent (377/774) of cases with an identifiable occupational group would be considered high risk for Q fever. The most common identifiable occupational group was agricultural/farming (31%). For the same period, at-risk environmental exposures were identified in 82% (961/1,170) of notifications; at-risk animal-related exposures were identified in 52% (612/1,170) of notifications; abattoir exposure was identified in 7% of notifications. This study has shown that the improved follow-up of Q fever cases since 2012 has been effective in the identification of possible exposure pathways for Q fever transmission. This improved surveillance has highlighted the need for further education and heightened awareness of Q fever risk for all people living in Queensland, not just those in previously-considered high risk occupations.

Background and Objective: Respiratory syncytial virus (RSV) is the most significant cause of acute respiratory infection (ARI) in early life. RSV and other respiratory viruses are known to stimulate substantial outgrowth of potentially pathogenic bacteria in the upper airways of young children. However, the clinical significance of interactions between viruses and bacteria is currently unclear. The present study aimed to clarify the effect of viral and bacterial co-detections on disease severity during paediatric ARI. Methods: Nasopharyngeal aspirates from children under 2years of age presenting with ARI to the emergency department were screened by quantitative PCR for 17 respiratory viruses and the bacterial pathogens Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis. Associations between pathogen detection and clinical measures of disease severity were investigated. Results: RSV was the most common virus detected, present in 29 of 58 samples from children with ARI (50%). Detection of S. pneumoniae was significantly more frequent during RSV infections compared to other respiratory viruses (adjusted effect size: 1.8, P: 0.03), and co-detection of both pathogens was associated with higher clinical disease severity scores (adjusted effect size: 1.2, P: 0.03). Conclusion: Co-detection of RSV and S. pneumoniae in the nasopharynx was associated with more severe ARI, suggesting that S. pneumoniae colonization plays a pathogenic role in young children.

International audience; Together with influenza, the non-influenza RNA respiratory viruses (NIRVs), which include respiratory syncytial virus, parainfluenza viruses, coronavirus, rhinovirus, and human metapneumovirus, represent a considerable global health burden, as recognised by WHO's Battle against Respiratory Viruses initiative. By contrast with influenza viruses, little is known about the contemporaneous global diversity of these viruses, and the relevance of such for development of pharmaceutical interventions. Although far less advanced than for influenza, antiviral drugs and vaccines are in different stages of development for several of these viruses, but no interventions have been licensed. This scarcity of global genetic data represents a substantial knowledge gap and impediment to the eventual licensing of new antiviral drugs and vaccines for NIRVs. Enhanced genetic surveillance will assist and boost research and development into new antiviral drugs and vaccines for these viruses. Additionally, understanding the global diversity of respiratory viruses is also part of emerging disease preparedness, because non-human coronaviruses and paramyxoviruses have been listed as priority concerns in a recent WHO research and development blueprint initiative for emerging infectious diseases. In this Personal View, we explain further the rationale for expanding the genetic database of NIRVs and emphasise the need for greater investment in this area of research.

An accurate rotavirus diagnosis is important for clinical management and monitoring active disease and vaccine effectiveness. Between 2016–2018, rotavirus-positive results in our laboratory were from vaccine virus shedding in 71/152 (46.7%) infants with a request for rotavirus testing. Routine infant diagnostic testing should ideally distinguish vaccine from wild-type viruses.

Human parechovirus (HPeV) is an emerging pathogen for infants. Improved diagnostics are needed due to the non-specific clinical presentation. Real-time reverse transcription polymerase chain reaction (RT-PCR) on blood samples may be an adjunct to diagnosis. A retrospective cohort of HPeV-affected infants was used to assess sensitivity and specificity of a HPeV RT-PCR on blood and cerebrospinal fluid (CSF). As a secondary analysis, the Ct value of the PCR results was compared to clinical correlates of severity. Between 2017 and 2018 blood samples were obtained from 97 infants of whom 44 had HPeV clinical and laboratory proven infection. Eighty-three concurrent CSF samples were available. Sensitivity was 93.3% [95% CI 82-99] for blood HPeV RT-PCR and 85% [95% CI 73.9-96.1] for CSF HPeV RT-PCR. Blood HPeV RT-PCR Ct values < 25 cycles were associated with age < 28 days and < 3 days of symptoms. No statistical associations were identified between potential clinical markers of severity and Ct value. HPeV RT-PCR on blood is a valuable adjunct to diagnostic testing for acute HPeV-related illness in infants. Results can be expected to be robust until at least day 5 of symptoms, with optimal sampling occurring close to onset of symptoms.

BackgroundHospital-based studies identify parechovirus (PeV), primarily PeV-A3, as an important cause of severe infections in young children. However, few community-based studies have been published and the true PeV infection burden is unknown. We investigated PeV epidemiology in healthy children participating in a community-based, longitudinal birth cohort study.MethodsAustralian children (n = 158) enrolled in the Observational Research in Childhood Infectious Diseases (ORChID) study were followed from birth until their second birthday. Weekly stool and nasal swabs and daily symptom diaries were collected. Swabs were tested for PeV by reverse-transcription polymerase chain reaction and genotypes determined by subgenomic sequencing. Incidence rate, infection characteristics, clinical associations, and virus codetections were investigated.ResultsPeV was detected in 1423 of 11 124 (12.8%) and 17 of 8100 (0.2%) stool and nasal swabs, respectively. Major genotypes among the 306 infection episodes identified were PeV-A1 (47.9%), PeV-A6 (20.1%), and PeV-A3 (18.3%). The incidence rate was 144 episodes (95% confidence interval, 128–160) per 100 child-years. First infections appeared at a median age of 8 (interquartile range, 6.0–11.7) months. Annual seasonal peaks changing from PeV-A1 to PeV-A3 were observed. Infection was positively associated with age ≥6 months, summer season, nonexclusive breastfeeding at age ConclusionsIn contrast with hospital-based studies, this study showed that diverse and dynamically changing PeV genotypes circulate in the community causing mild or subclinical infections in children.Parechovirus can cause severe illnesses in children. However, studies focus mainly on hospitalized populations. True disease burden in the community remains largely unknown. From our community-based cohort, we found diverse parechovirus genotypes in the community, causing mild or subclinical infections in children.Clinical Trials RegistrationNCT01304914.

Background: Hospital-based studies identify human parechovirus (HPeV), primarily HPeV3, as important causes of severe infections in young children. However, few community-based studies have been published and the true HPeV infection burden is unknown. We therefore investigated HPeV epidemiology in healthy children participating in a community-based, longitudinal birth cohort study. Methods: Australian children (n=158) enrolled in the Observational Research in Childhood Infectious Diseases (ORChID) study were followed from birth until their second birthday. Weekly stool and nasal swabs, and daily symptom diaries, were collected. Swabs were tested for HPeV by reverse-transcription polymerase chain reaction and genotypes determined by subgenomic sequencing. Incidence rate, infection characteristics, clinical associations, and virus co-detections were investigated. Findings: HPeV was detected in 1423/11127 (12·8%) and 17/8100 (0·2%) stool and nasal swabs, respectively. Major genotypes amongst the 306 infection episodes identified were HPeV1 (47·9%), HPeV6 (20·1%), and HPeV3 (18·3%). The incidence rate was 144 episodes/100 child-years (95% CI 128-160). First infections appeared at a median age of 8·0-months (interquartile range 6·0-11·7). Annual seasonal peaks changing from HPeV1 to HPeV3 were observed. Infection was positively associated with age ≥6-months, summer season, non-exclusive breastfeeding at age

There is limited knowledge of the true prevalence and distribution of coxiellosis in dairy and beef cattle populations in Australia. For this to occur, apparent prevalence estimates need to be reliably adjusted, accounting for diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the test used. However, there are few tests available with known diagnostic specifications suitable to inform screening and surveillance activities in the Australian context. We initially modified and optimised a human indirect immunofluorescence assay (IFA) test for the detection of IgG antibodies against phase I and/or phase II Coxiella burnetii in bovine sera and determined an optimal screening dilution cut-off to be 1:160. Direct comparison of the modified IFA with the commercial IDEXX enzyme-linked immunosorbent assay (ELISA) kit (Q Fever Ab Test IDEXX Laboratories, United States of America) was performed by testing 458 serum samples from four distinct cattle populations across the east coast of Australia and New Zealand. Cross classified test results were then analysed using Bayesian latent class modelling, to validate the tests in the absence of a gold standard reference test. Results from this analysis indicate that the IFA, at a 1:160 serum dilution, has an estimated DSe of 73.6% (95% Credible Interval (CrI) 61.1, 85.9) and DSp of 98.2% (95% CrI 95.1, 99.7). The commercial IDEXX ELISA kit was found to have a higher DSe of 87.9% (95% CrI 73.9, 96.4) and similar DSp of 97.7% (95% CrI 93.2, 99.7). Evaluation of the diagnostic performance of the IFA and ELISA methods, specifically for use in cattle will enable more accurate interpretation of prevalence estimates of C. burnetii exposure to be reported for cattle in Australia and other countries.

AIM The human parechovirus (HPeV) has emerged as a pathogen causing sepsis-like presentations in young infants, but there is a lack of data on HPeV presentations requiring intensive care support. We aimed to characterise the clinical presentation, disease severity, management and outcome of a population-based cohort of children with microbiologically confirmed HPeV infection requiring admission to paediatric intensive care units (PICUs) in Queensland, Australia during a recent outbreak. METHODS This was a multicentre retrospective study of children admitted to PICU between 1 January 2015 and 31 December 2016 with confirmed HPeV infection. RESULTS Thirty infants (median age 20 days) with HPeV genotype 3 were admitted to PICU, representing 16% of all children with HPeV admitted to hospital and 6.4% of non-elective PICU admissions in children

Coxiella burnetii is the causative agent of the zoonotic disease, Q fever. The disease Q fever is diagnosed globally, however Australia, and in particular Queensland, has the highest rates of notifications in the world. This study is the first comprehensive investigation of Q fever in Queensland, with a special focus on paediatric infections. At the time of commencement, very limited data were available on the incidence of Q fever in Queensland, the risk factors associated with infection, and the presentation of the disease in children. Analysis of 6,797 notified Queensland Q fever cases, confirmed the disease to be mainly confined to rural populations, yet there were a noteworthy number of cases reported from urban and non-rural communities. Alarmingly there were 235 notifications from children in Queensland. Globally, Q fever is considered to be under-reported and therefore a seroprevalence study was performed to truly assess the rate of exposure in the Queensland population. This seroprevalence study highlighted a wider exposure to the organism, with a substantial number of people being exposed from non-rural communities, along with an increased prevalence in children. Further investigations were performed to identify the sources of exposure, especially in “low risk” populations using PCR. This study identified a number of potential sources of infection to humans including domestic pets, flying foxes and dust. Q fever is an infectious disease presenting with a wide variety of symptoms which may obscure the clinician’s diagnostic approach and treatment. This thesis investigated specific cases of Q fever in both children and adults where there were unusual presentations, including whole families infected, and a serve case in which the patient had complete organ failure. These cases showed the need for greater understanding and awareness of the disease and also demonstrated that indirect transmission of infection from a family member working in a high risk occupation, to other family members may occur. Prevention of Q fever in the population through vaccination is highly desirable. To ensure adequate vaccine coverage, genotypic characterisation of the Coxiella strains circulating in the population is necessary. This study determined the range of different genotypes detected, and found new, novel genotypes circulating in Queensland, as well as genotypes that were unique to Australia. Q fever is a vaccine preventable disease in Australia and has had Federal Government funding to protect workers in high risk occupations. Yet the data generated in this thesis highlighted the need for the vaccine strategy to be revised so that children and family, members of at risk workers are included. The current vaccine has many limitations, including that it requires extensive pre-screening procedures to prevent previously exposed subjects from having an adverse reaction upon vaccination. The vaccine can induce a hyper-sensitised reaction in some subjects, eliciting symptoms of Q fever. As part of this thesis a cell mediated immunity assay was developed to enable vaccine candidates to be screened for previous exposure without having to be re-exposed to the bacteria in the widely applied skin test. This pre-screening tool will allow children to be screened without the potential for an adverse hypersensitive reaction to occur, and should facilitate the wider administration of the vaccine to this younger age group. In summary, the results presented in this Thesis, address the knowledge gap regarding the epidemiology and clinical impact of Q fever in the Queensland population. It presents evidence of other populations at risk, which may help to formulate an improved vaccination strategy for Queenslanders, and help shape the Public Health approach to the management of this serious disease.

Q fever is a vaccine preventable disease; however, despite this, high notification numbers are still recorded annually in Australia. We investigated the seroprevalence of Coxiella burnetii, the Q fever agent, in a Queensland sample population. Notification data (N = 6425) from 1984-2008 were collated, identifying high risk areas of Q fever exposure. Of these 177 were recorded in children. Serum samples were collected from Queensland and screened using both an immunoflourescence assay at 1:10 dilution and a commercially available ELISA kit. Results were collated based on age, geographical location and sex. From 1988 Queensland samples screened, 103 were identified as Q fever IgG-positive, giving a seroprevalence of 5.2% (95% CI 4.3-6.2%). Seroprevalence in the rural/remote population was 5.3% (95% CI 4.6-6.6%), and the metropolitan Brisbane population, which is considered not at risk, was 5.0% (95% CI 3.7-6.7%). Sixty-three seropositive males and 40 females were identified, along with an increase in seropositivity with increasing age. The seropositivity of children was 1.3% (95% CI 0.7-2.3%) from 844 samples. We have shown that both metropolitan and paediatric populations which are considered low risk of Coxiella exposure have surprisingly high seropositivity. These emerging groups require further investigation and consideration for the introduction of preventive measures.

The post-marketing undesirable events to hair colouring products in the European Union notified to the cosmetovigilance departments of four major cosmetic companies were analysed (2003-2006). The objective was to determine whether there was any time effect (trend to increase or decrease), country effect (significant difference between the countries included in the analysis) or product type effect (direct or oxidation), as well as to identify risk factors. Alleged undesirable events (UEvs, all notifications prior to causality assessment), were compared to the respective undesirable effects (UEfs, reasonably attributable to product use). A detailed analysis was performed on notifications with manifestations compatible with allergic contact dermatitis. No time effect of UEvs and UEfs was shown, for all hair-dye associated notifications and for allergic contact dermatitis, for all hair colouring products together and by product type. The incidence of allergic contact dermatitis to direct hair colouring products was lower for all four companies compared to oxidative hair dyes. The reporting rates of UEfs were statistically higher in the UK for one of four companies. Past history of black henna tattoos appeared as a major risk factor for seriousness of allergic contact reactions.

The polyomavirus family is rapidly expanding with twelve new human viruses identified since 2007. A significant number of the new human polyomaviruses (HPyV) has been found on the skin. Whether these viruses share biological properties and should be grouped together is unknown. Here we investigated the serological behavior of cutaneous HPyVs in a general population. 799 sera from immunocompetent Australian individuals aged between 0-87 were analyzed with a Luminex xMAP technology-based immunoassay for the presence of VP1-directed IgG antibodies against MCPyV, HPyV6, HPyV7, TSPyV, HPyV9, and BKPyV as a control. Except for HPyV9, overall seropositivity was high for the cutanous polyomaviruses (66-81% in adults), and gradually increased with age. Children below 6 months displayed seropositivity rates comparable to the adults, indicative of maternal antibodies. TSPyV seroreactivity levels strongly increased after age 2 and waned later in life comparable to BKPyV, whereas MCPyV, HPyV6 and HPyV7 seroreactivity remained rather stable throughout. Based on the identified serologic profiles, MCPyV seems to cluster with HPyV6 and HPyV7, and TSPyV and HPyV9 by themselves. These profiles indicate heterogeneity among cutaneous polyomaviruses and probably reflect differences in exposure and pathogenic behavior of these viruses.

Q fever is a vaccine-preventable disease; despite this, high annual notification numbers are still recorded in Australia. We have previously shown seroprevalence in Queensland metropolitan regions is approaching that of rural areas. This study investigated the presence of nucleic acid from Coxiella burnetii, the agent responsible for Q fever, in a number of animal and environmental samples collected throughout Queensland, to identify potential sources of human infection. Samples were collected from 129 geographical locations and included urine, faeces and whole blood from 22 different animal species; 45 ticks were removed from two species, canines and possums; 151 soil samples; 72 atmospheric dust samples collected from two locations and 50 dust swabs collected from domestic vacuum cleaners. PCR testing was performed targeting the IS1111 and COM1 genes for the specific detection of C. burnetii DNA. There were 85 detections from 1318 animal samples, giving a detection rate for each sample type ranging from 2.1 to 6.8%. Equine samples produced a detection rate of 11.9%, whilst feline and canine samples showed detection rates of 7.8% and 5.2%, respectively. Native animals had varying detection rates: pooled urines from flying foxes had 7.8%, whilst koalas had 5.1%, and 6.7% of ticks screened were positive. The soil and dust samples showed the presence of C. burnetii DNA ranging from 2.0 to 6.9%, respectively. These data show that specimens from a variety of animal species and the general environment provide a number of potential sources for C. burnetii infections of humans living in Queensland. These previously unrecognized sources may account for the high seroprevalence rates seen in putative low-risk communities, including Q fever patients with no direct animal contact and those subjects living in a low-risk urban environment.

Neisseria meningitidis is the causative agent of invasive meningococcal disease (IMD). The BEXSERO® vaccine which is used to prevent serogroup B disease is composed of four sub-capsular protein antigens supplemented with an outer membrane vesicle. Since the sub-capsular protein antigens are variably expressed and antigenically variable amongst meningococcal isolates, vaccine coverage can be estimated by the meningococcal antigen typing system (MATS) which measures the propensity of the strain to be killed by vaccinated sera. Whole genome sequencing (WGS) which identifies the alleles of the antigens that may be recognised by the antibody response could represent, in future, an alternative estimate of coverage. In this study, WGS of 278 meningococcal isolates responsible for 62% of IMD in Western Australia from 2000–2014 were analysed for association of genetic lineage (sequence type [ST], clonal complex [cc]) with BEXSERO® antigen sequence type (BAST) and MATS to predict the annual vaccine coverage. A hyper-endemic period of IMD between 2000–05 was caused by cc41/44 with the major sequence type of ST-146 which was not predicted by MATS or BAST to be covered by the vaccine. An increase in serogroup diversity was observed between 2010–14 with the emergence of cc11 serogroup W in the adolescent population and cc23 serogroup Y in the elderly. BASTs were statistically associated with clonal complex although individual antigens underwent antigenic drift from the major type. BAST and MATS predicted an annual range of 44–91% vaccine coverage. Periods of low vaccine coverage in years post-2005 were not a result of the resurgence of cc41/44:ST-146 but were characterised by increased diversity of clonal complexes expressing BASTs which were not predicted by MATS to be covered by the vaccine. The driving force behind the diversity of the clonal complex and BAST during these periods of low vaccine coverage is unknown, but could be due to immune selection and inter-strain competition with carriage of non-disease causing meningococci.

Human bocavirus (HBoV) has been detected worldwide in respiratory samples. Two real-time PCR assays, targeting the non-structural protein (NP-1) and viral protein (VP-1) genes, were designed and validated to detect HBoV in patients with respiratory disease, gastroenteritis, or systemic illness. Sensitivity of the NP-1 and VP-1 assays were equal to the conventional PCR assay previously described by Allander et al. [2005: Proc Natl Acad Sci USA 102: 12891-12896] being 100%, and giving specificity of 94% and 93%, respectively. There was no cross-reaction identified with unrelated respiratory agents, or to human DNA. The limits of detection were 10 copies of genomic DNA equivalents per reaction for both assays. The assays were used to screen three different sample populations, combined nose, and throat swabs (n = 96) from children with acute respiratory disease, fecal samples (n = 375) from adults, and children with gastroenteritis and whole blood (n = 229) collected from 31 immunocompromised children taken over an 18-month period. In total 17 (18%) respiratory samples and 18 (4.8%) fecal samples were identified as having HBoV present. Of the pediatric whole blood specimens investigated, HBoV was detected in six (2.6%) samples from four patients. In summary, two real-time PCR assays targeting different genes were designed and validated for use as screening methods for the detection of HBoV. HBoV was found in three different specimen types: parent-collected combined nose-throat swabs, fecal samples collected from symptomatic individuals and whole blood from immunocompromised children.

A 28-year-old woman, a park ranger, developed acute Q fever with associated sepsis, profound jaundice, disseminated intravascular coagulation and multiorgan failure necessitating prolonged admission to the intensive care unit for ventilatory support. She recovered fully and remains well 4 years later.

Background The accurate quantification of Plasmodium falciparum parasite numbers by PCR is an important tool for monitoring growth kinetics in subjects infected and subsequently treated with anti-malarial agents. Methods A real-time quantitative PCR (rt-qPCR) method using primers and a hydrolysis probe that targets the 18S rRNA gene was adapted and optimized to estimate parasite load in blood samples. Samples included laboratory prepared blood samples of varying parasite concentrations (6.4 × 105 to 6.4 parasites per 500 μl of packed red blood cells (500pRBC)) and blood samples collected from an experimentally infected human subject collected at 19 time points over 10 days. Sample preparation and extraction, detection chemistry, assay reproducibility, and limit of detection were compared to a previously published SYBR Green rt-qPCR used in a malaria vaccine clinical trial. Results Both the rt-qPCR hydrolysis probe assay and SYBR Green rt-qPCR provided a limit of detection of 6.4 × 101 parasites per 500pRBC. However non-specific amplification in the SYBR Green rt-qPCR assay led to either inaccurate estimation of parasite load at levels below 6.4 × 102 parasites per 500pRBC and to false-positive detection of parasites in negative samples. The rt-qPCR hydrolysis probe assay was specific and provided reliable quantification of parasitaemia down to 6.4 × 101 parasites per 500pRBC. Notably, 12 of the 19 consecutive samples collected from the experimentally infected subject were at or below 6.4 × 102 copies per 500pRBC. Conclusions These results show that the hydrolysis probe rt-qPCR assay is superior to the SYBR Green rt-qPCR for the quantification of P. falciparum in human blood samples. The hydrolysis probe rt-qPCR is now in use in the Queensland paediatric infectious diseases laboratory (QPID) to monitor parasitaemia in experimentally-infected clinical trial subjects.

In the last two decades, sustainable and safe use of chemicals became a focal point of various international policy efforts and has been addressed through launching of chemical management systems worldwide. In the EU, the regulation on Registration, Evaluation and Authorization of Chemicals (REACH) primarily concentrates on generating substance intrinsic properties data (including hazard) depending on the tonnage being registered. However, using the chemical production/import volume alone as a proxy/indicator of true exposures undermines risk management decision making and deprives exposure information of its real power (e.g. smart studies design, more efficient prioritization). The European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) together with the European Chemical Industry Council Long Range Initiative program streamline their research efforts towards gaining wider regulatory acceptance of exposure-informed testing and risk management practices. This presentation provides an overview of recent industry-funded research building a knowledgebase on tools and methods for assessment of low exposure. The problem definition of low exposure in various contexts will be discussed, focusing on aspects/factors that make low exposure very important in certain situations. A detailed outline will be provided for multiple projects that consider low exposure from differentperspectives. The examples will cover comparative exposure analysis of alternative exposure sources to the same substance, assessment of intentional and non-intentional exposure, exposure based waiving and workplace risk assessment. The presented material should inform the workshop participants about the ECETOC’s perspective on the importance/weight of (low) exposure in regulatory risk assessment and set the stage for a follow-up panel discussion to help identify remaining knowledge gaps and future focus areas.