Salmonella source attribution in a subtropical state of Australia: capturing environmental reservoirs of infection.

Salmonellosis is a leading cause of hospitalisation due to gastroenteritis in Australia. A previous source attribution analysis for a temperate state in Australia attributed most infections to chicken meat or eggs. Queensland is in northern Australia and includes subtropical and tropical climate zones. We analysed Queensland notifications for salmonellosis and conducted source attribution to compare reservoir sources with those in southern Australia. In contrast to temperate Australia, most infections were due to non-Typhimurium serotypes, with particularly high incidence in children under 5 years and strong seasonality, peaking in summer. We attributed 65.3% (95% credible interval (CrI) 60.6-73.2) of cases to either chicken meat or eggs and 15.5% (95% CrI 7.0-19.5) to nuts. The subtypes with the strongest associations with nuts were Salmonella Aberdeen, S. Birkenhead, S. Hvittingfoss, S. Potsdam and S. Waycross. All five subtypes had high rates of illness in children under 5 years (ranging from 4/100 000 to 23/100 000), suggesting that nuts may be serving as a proxy for environmental transmission in the model. Australia's climatic range allows us to conduct source attribution in different climate zones with similar food consumption patterns. This attribution provides evidence for environment-mediated transmission of salmonellosis in sub-tropical regions.

A population-based longitudinal study of Clostridium difficile infection-related hospitalization in mid-age and older Australians.

Clostridium difficile is the principal cause of infectious diarrhoea in hospitalized patients. We investigated the incidence and risk factors for hospitalization due to C. difficile infection (CDI) in older Australians. We linked data from a population-based prospective cohort study (the 45 and Up Study) of 266 922 adults aged ⩾45 years recruited in New South Wales, Australia to hospitalization and death records for 2006-2012. We estimated the incidence of CDI hospitalization and calculated days in hospital and costs per hospitalization. We also estimated hazard ratios (HR) for CDI hospitalization using Cox regression with age as the underlying time variable. Over a total follow-up of 1 126 708 person-years, 187 adults had an incident CDI hospitalization. The crude incidence of CDI hospitalization was 16·6/100 000 person-years, with a median hospital stay of 6 days, and a median cost of AUD 6102 per admission. Incidence increased with age and year of follow-up, with a threefold increase for 2009-2012. After adjustment, CDI hospitalization rates were significantly lower in males than females (adjusted HR 0·6, 95% confidence interval 0·4-0·7). CDI hospitalization rates increased significantly over 2009-2012. There is a need to better understand the increasing risk of CDI hospitalization in women.

The campylobacteriosis conundrum - examining the incidence of infection with Campylobacter sp. in Australia, 1998-2013.

Campylobacter sp. are a globally significant cause of gastroenteritis. Although rates of infection in Australia are among the highest in the industrialized world, studies describing campylobacteriosis incidence in Australia are lacking. Using national disease notification data between 1998 and 2013 we examined Campylobacter infections by gender, age group, season and state and territory. Negative binomial regression was used to estimate incidence rate ratios (IRRs), including trends by age group over time, with post-estimation commands used to obtain adjusted incidence rates. The incidence rate for males was significantly higher than for females [IRR 1·20, 95% confidence interval (CI) 1·18-1·21], while a distinct seasonality was demonstrated with higher rates in both spring (IRR 1·18, 95% CI 1·16-1·20) and summer (IRR 1·17, 95% CI 1·16-1·19). Examination of trends in age-specific incidence over time showed declines in incidence in those aged <40 years combined with contemporaneous increases in older age groups, notably those aged 70-79 years (IRR 1998-2013: 1·75, 95% CI 1·63-1·88). While crude rates continue to be highest in children, our findings suggest the age structure for campylobacteriosis in Australia is changing, carrying significant public health implications for older Australians.

A large outbreak of Campylobacter jejuni infection in a university college caused by chicken liver pâté, Australia, 2013.

In October 2013, public health authorities were notified of a suspected outbreak of gastroenteritis in students and guests following a catered function at a university residential college. A retrospective cohort study was undertaken to examine whether foods served at the function caused illness. A total of 56 cases of gastroenteritis, including seven laboratory-confirmed cases of Campylobacter jejuni infection, were identified in 235 eligible respondents. Univariate analysis showed a significant association with a chicken liver pâté entrée [relative risk (RR) 3·64, 95% confidence interval (CI) 2·03-6·52, P < 0·001], which retained significance after adjustment for confounding via multivariable analysis (adjusted RR 2·80, 95% CI 1·26-6·19, P = 0·01). C. jejuni and C. coli were also isolated in chicken liver pâté recovered from the college's kitchen. Subsequent whole genome multilocus sequence typing (wgMLST) of clinical and food-derived C. jejuni isolates showed three genetically distinct sequence types (STs) comprising ST528, ST535 (both clinically derived) and ST991 (food derived). The study demonstrates the value of utilizing complementary sources of evidence, including genomic data, to support public health investigations. The use of wgMLST highlights the potential for significant C. jejuni diversity in epidemiologically related human and food isolates recovered during outbreaks linked to poultry liver.

Evaluation of a structured expert elicitation estimating the proportion of illness acquired by foodborne transmission for nine enteric pathogens in Australia.

Estimates of the proportion of illness transmitted by food for different enteric pathogens are essential for foodborne burden-of-disease studies. Owing to insufficient scientific data, a formal synthesis of expert opinion, an expert elicitation, is commonly used to produce such estimates. Eleven experts participated in an elicitation to estimate the proportion of illnesses due to food in Australia for nine pathogens over three rounds: first, based on their own knowledge alone; second, after being provided with systematic reviews of the literature and Australian data; and finally, at a workshop where experts reflected on the evidence. Estimates changed significantly across the three rounds (P = 0·002) as measured by analysis of variance. Following the workshop in round 3, estimates showed smoother distributions with significantly less variation for several pathogens. When estimates were combined to provide combined distributions for each pathogen, the width of these combined distributions reflected experts' perceptions of the availability of evidence, with narrower intervals for pathogens for which evidence was judged to be strongest. Our findings show that the choice of expert elicitation process can significantly influence final estimates. Our structured process - and the workshop in particular - produced robust estimates and distributions appropriate for inclusion in burden-of-disease studies.

Bayesian Source Attribution of Salmonellosis in South Australia.

Salmonellosis is a significant cause of foodborne gastroenteritis in Australia, and rates of illness have increased over recent years. We adopt a Bayesian source attribution model to estimate the contribution of different animal reservoirs to illness due to Salmonella spp. in South Australia between 2000 and 2010, together with 95% credible intervals (CrI). We excluded known travel associated cases and those of rare subtypes (fewer than 20 human cases or fewer than 10 isolates from included sources over the 11-year period), and the remaining 76% of cases were classified as sporadic or outbreak associated. Source-related parameters were included to allow for different handling and consumption practices. We attributed 35% (95% CrI: 20-49) of sporadic cases to chicken meat and 37% (95% CrI: 23-53) of sporadic cases to eggs. Of outbreak-related cases, 33% (95% CrI: 20-62) were attributed to chicken meat and 59% (95% CrI: 29-75) to eggs. A comparison of alternative model assumptions indicated that biases due to possible clustering of samples from sources had relatively minor effects on these estimates. Analysis of source-related parameters showed higher risk of illness from contaminated eggs than from contaminated chicken meat, suggesting that consumption and handling practices potentially play a bigger role in illness due to eggs, considering low Salmonella prevalence on eggs. Our results strengthen the evidence that eggs and chicken meat are important vehicles for salmonellosis in South Australia.

Incidence of acute respiratory infections in Australia.

We used a national survey of 7578 randomly selected respondents in 2008-2009 to identify the period prevalence of acute respiratory infection (ARI) by season and state, and to estimate the incidence of ARI in the Australian community. A case was defined as any episode of cold or flu with at least one of the following symptoms: fever, chills, sore throat, running nose, or cough in the past 4 weeks. Frequency data were weighted to the Australian population. The response rate to the survey was 49%, and 19·9% (1505/7578) of respondents reported an ARI in the previous 4 weeks, which extrapolated to 68·9 million cases [95% confidence interval (CI) 65·1-72·7] of ARI in Australia annually. The incidence was 3·2 (95% CI 3·0-3·4) cases of ARI/person per year, and was highest in young children and lowest in older people. ARI imposes a significant burden on Australian society.

Relative rates of cancers and deaths in Australian communities with PFAS environmental contamination associated with firefighting foams: A cohort study using linked data.

BACKGROUND: Per- and polyfluoroalkyl substances (PFAS) are environmental contaminants that are potentially harmful to health. We examined if rates of selected cancers and causes of deaths were elevated in three Australian communities with local environmental contamination caused by firefighting foams containing PFAS. The affected Australian communities were Katherine in Northern Territory, Oakey in Queensland and Williamtown in New South Wales. METHODS: All residents identified in the Medicare Enrolment File (1983-2019)-a consumer directory for Australia's universal healthcare-who ever lived in an exposure area (Katherine, Oakey and Williamtown), and a sample of those who ever lived in selected comparison areas, were linked to the Australian Cancer Database (1982-2017) and National Death Index (1980-2019). We estimated standardised incidence ratios (SIRs) for 23 cancer outcomes, four causes of death and three control outcomes, adjusting for sex, age and calendar time of diagnosis. FINDINGS: We observed higher rates of prostate cancer (SIR=1·76, 95 % confidence interval (CI) 1·36-2·24) in Katherine; laryngeal cancer (SIR=2·71, 95 % CI 1·30-4·98), kidney cancer (SIR=1·82, 95 % CI 1·04-2·96) and coronary heart disease (CHD) mortality (SIR=1·81, 95 % CI 1·46-2·33) in Oakey; and lung cancer (SIR=1·83, 95 % CI 1·39-2·38) and CHD mortality (SIR=1·22, 95 % CI 1·01-1·47) in Williamtown. We also saw elevated SIRs for control outcomes. SIRs for all other outcomes and overall cancer were similar across exposure and comparison areas. INTERPRETATION: There was limited evidence to support an association between living in a PFAS exposure area and risks of cancers or cause-specific deaths.

Gastroenteritis in older people living in the community: results of two Australian surveys.

We analysed two large national surveys conducted in 2001 and 2008 to examine incidence and outcomes of gastroenteritis in older Australians. A case was someone reporting ≥3 loose stools or ≥1 episode of vomiting in 24 h, excluding non-infectious causes. We compared cases arising in the elderly (≥65 years) and in other adults (20-64 years). Elderly people experienced 0·33 [95% confidence interval (CI) 0·24-0·42] episodes of gastroenteritis/person per year, compared to 0·95 (95% CI 0·74-1·15) in other adults. Elderly cases reported less stomach cramps, fever and myalgia than younger cases, and were more likely to be hospitalized, although this was not statistically significant. In multivariable analysis, gastroenteritis in elderly people was associated with travelling within the state (odds ratio 1·35, 95% CI 1·07-1·71). Elderly people were less concerned about food safety than other adults. Older Australians were less likely to report gastroenteritis and experienced different symptoms and outcomes from other adults.

Serum concentration trends and apparent half-lives of per- and polyfluoroalkyl substances (PFAS) in Australian firefighters.

BACKGROUND: Per- and polyfluoroalkyl substances (PFASs) are persistent manmade compounds used in aqueous film forming foam (AFFF). The extensive use of AFFF has led to widespread environmental PFAS contamination and exposures of firefighters. OBJECTIVES: To determine PFAS blood serum concentration trends and apparent serum half-lives in firefighters after the replacement of AFFF. METHODS: Current and former employees of an Australian corporation providing firefighting services, where AFFF formulations had been used since the 1980s up until 2010, were recruited in 2018-2019 to participate in this study. Special focus was put on re-recruiting participants who had provided blood samples five years prior (2013-2014). Participants were asked to provide a blood sample and fill in a questionnaire. Serum samples were analysed for 40 different PFASs using HP LC-MS/MS. RESULTS: A total of 799 participants provided blood samples in 2018-2019. Of these, 130 previously provided blood serum in 2013-2014. In 2018-2019, mean (arithmetic) serum concentrations of perfluorooctane sulfonate (PFOS, 27 ng/mL), perfluoroheptane sulfonate (PFHpS, 1.7 ng/mL) and perfluorohexane sulfonate (PFHxS, 14 ng/mL) were higher than the levels in the general Australian population. Serum concentrations were associated with the use of PFOS/PFHxS based AFFF. Participants who commenced service after the replacement of this foam had serum concentrations similar to those in the general population. Mean (arithmetic) individual apparent half-lives were estimated to be 5.0 years (perfluorooctanoic acid (PFOA)), 7.8 years (PFHxS), 7.4 years (PFHpS) and 6.5 years (PFOS). CONCLUSION: This study shows how workplace interventions such as replacement of AFFF can benefit employees at risk of occupational exposure.

A national case-control study of risk factors for listeriosis in Australia.

Listeriosis is a foodborne disease associated with significant mortality. This study attempts to identify risk factors for sporadic listeriosis in Australia. Information on underlying illnesses was obtained from cases' treating doctors and other risk factors were elicited from the patient or a surrogate. We attempted to recruit two controls per case matched on age and primary underlying immune condition. Between November 2001 and December 2004 we recruited 136 cases and 97 controls. Of perinatal cases, living in a household where a language other than English was spoken was the main risk factor associated with listeriosis (OR 11·3, 95% CI 1·5-undefined). Of non-perinatal cases we identified the following risk factors for listeriosis: prior hospitalization (OR 4·3, 95% CI 1·0-18·3), use of gastric acid inhibitors (OR 9·4, 95% CI 2·4-37·4), and consumption of camembert (OR 4·7, 95% CI 1·1-20·6). Forty percent of cases with prior hospitalization were exposed to high-risk foods during hospitalization.

Identified interneurons produce both primary afferent depolarization and presynaptic inhibition.

Crayfish interneurons were identified that appear to be directly responsible for presynaptic inhibition of primary afferent synapses during crayfish escape behavior. The interneurons are fired by a polysynaptic pathway triggered by the giant escape command axons. When directly stimulated, these interneurons produce short-latency, chloride-dependent primary afferent depolarizations and presynaptically inhibit primary afferent input to mechanosensory interneurons.

The Origin of the Superstructure in Bi2Sr2CaCu2O8+dgr as Revealed by Scanning Tunneling Microscopy.

Real-space images with atomic resolution of the BiO plane of Bi(2)Sr(2)CaCu(2)O(8+delta) were obtained with a scanning tunneling microscope. Single-crystal samples were cleaved and imaged under ultrahigh vacuum conditions at room temperature. The images clearly show the one-dimensional incommensurate superstructure along the b-axis that is common to this phase. High-resolution images show the position of the Bi atoms, revealing the structural nature of the superlattice. A missing row of Bi atoms occurs either every nine or ten atomic sites in both (110) directions, accounting for the measured incommensurate periodicity of the superstructure. A model is proposed that includes missing rows of atoms, as well as displacements of the atomic positions along both the a- and c-axis directions.

Higher rate of culture-confirmed Campylobacter infections in Australia than in the USA: is this due to differences in healthcare-seeking behaviour or stool culture frequency?

Laboratory-based surveillance by OzFoodNet in Australia and FoodNet in the USA indicated that the incidence of Campylobacter infections in 2001 in Australia was about nine times higher than in the USA. We assessed whether this disparity could be explained by differences in the frequency of stool culturing. Using data from population surveys of diarrhoea and symptom profiles for Campylobacter from case-control studies, indices of healthcare behaviour taking into account the severity of Campylobacter infections were calculated. These suggest that culture-confirmed Campylobacter infections underestimate the incidence of community cases by similar ratios in the two countries. The incidence of Campylobacter infections in Australia was about 12 times higher than in the USA after consideration of healthcare system differences.

Per- and polyfluoroalkyl substances (PFAS) in Australia: Current levels and estimated population reference values for selected compounds.

BACKGROUND: Increased public awareness of PFAS contamination in Australia has resulted in serum biomonitoring efforts in individuals in potentially affected communities. However, population-based reference values for assessing whether individual results exceed the typical range in the Australian general population are not currently available. OBJECTIVE: Estimate population upper bound reference values based on updated serum PFAS concentrations in pooled samples from southeast Queensland, Australia and population variation observed in the US National Health and Nutrition Examination Survey (NHANES) datasets. METHODS: We calculated ratios of 95th percentile to arithmetic mean (P95:AM ratios) using data from the NHANES 2013-14 and 2015-16 cycle samples for frequently detected PFASs: PFOA, linear and branched PFOS, perfluorononanoate (PFNA), perfluorodecanoate (PFDA), and perfluorohexanesulfonate (PFHxS). We estimated Australian age-specific means for PFAS using pooled serum samples collected in 2014-15 and 2016-17. We used the P95:AM ratios to estimate 95th percentile concentrations in the Australian population based on the results of the 2016-17 pooled samples. RESULTS AND CONCLUSIONS: P95:AM ratios for each PFAS were similar across NHANES cycle and age group, so overall compound-specific ratios were estimated for PFOA (2.1), PFNA (2.4), PFDA (2.7), PFHxS (2.7), and linear (2.4) and summed PFOS (2.3). Australian mean PFAS concentrations continued previously reported declining trends. The estimated P95 values can be used as preliminary substitutes for more rigorous population reference values to identify samples with clearly elevated serum PFAS concentrations in Australian biomonitoring efforts. Given uncertainties and variability inherent in this evaluation, the estimated P95 values should be interpreted with caution. Mean and estimated P95 serum PFAS concentrations in Australia should continue to be monitored to document declining trends in population serum concentrations.

Internationally distributed frozen oyster meat causing multiple outbreaks of norovirus infection in Australia.

BACKGROUND: Between November 2003 and January 2004, outbreaks of norovirus in 3 Australian jurisdictions involving 83 cases of illness were associated with imported oyster meat. METHODS: Cohort studies were conducted in 2 jurisdictions to identify relative risks of illness for the consumption of oysters. A case series was conducted in the third jurisdiction. RESULTS: The cohort studies conducted in the first 2 jurisdictions identified relative risks of illness of 17 (95% confidence interval, 5-51) and 35 (95% confidence interval, 5-243), respectively, for the consumption of oysters. Multiple strains of norovirus were detected in fecal specimens from 8 of 14 patients and in 1 of the 3 batches of implicated oyster meat using seminested reverse-transcriptase polymerase chain reaction methods. Traceback investigations revealed that all oyster meat was harvested from the same estuary system in Japan within the same month. CONCLUSIONS: These outbreaks demonstrate the potential of foodborne disease to spread internationally and the need for national and international collaboration to investigate such outbreaks. Foodborne illness related to norovirus is underestimated because of underreporting of human cases and challenges in laboratory detection of viruses in foods, both of which can delay public health action.

Extracellular matrix synthesized by clonal osteogenic cells is osteoinductive in vivo and in vitro: role of transforming growth factor-beta 1 in osteoblast cell-matrix interaction.

ROB-C26 (C26) is a multipotential, clonal cell line known to express several members of the TGF-beta superfamily and to become more osteoblastic (e.g., express higher levels of alkaline phosphatase) upon treatment with 10(-6)M retinoic acid (RA). We hypothesize that the expression of this more osteoblastic phenotype subsequent to RA exposure is the result of the treated cell's extracellular matrix (ECM) becoming a repository and active source of putative osteoinductive growth factors including, specifically, select members of the TGF-beta superfamily. To test this hypothesis, we isolated the ECM from RA-treated and untreated C26 cells and assessed them for their ability to promote osteogenic differentiation in vivo and in vitro. We then explored whether the latter activities could be attributed specifically to TGF-beta 1. We found that the ECM of treated cells isolated by cell lysis and extensive washing induced endochondral bone formation in vivo when implanted into the thigh muscles of athymic nude mice and stimulated alkaline phosphatase (ALP) activity in vitro in freshly plated C26 cells. This latter stimulation was comparable to levels observed with direct RA treatment. This latter in vitro activity was only very partially mimicked by the ECM prepared from untreated cells and not duplicated at all by RA-treated collagen or the ECM from another RA-treated multipotential cell line. Moreover, the in vivo osteoinductive effect of the treated C26 cell ECM was not duplicated by comparable ECM prepared from untreated cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Cellular and synaptic morphology of a feeding motor circuit in Aplysia californica.

The cellular and synaptic morphology of a component of the feeding motor circuit in Aplysia californica was examined with light and electron microscopic techniques. The circuit consists of a pair of inhibitory premotor interneurons, B4 and B5, as well as two motoneurons, B15 and B16, which innervate the accessory radula closer muscle. The neurons have wide, varicose arborizations in the buccal ganglion neuropil. All four of these neurons are cholinergic, and in addition, B15 contains immunoreactivity to sera raised against small cardioactive peptide B. Varicose processes in the accessory radula closer muscle are immunoreactive with antisera against several neuropeptides. We identified specific neuromuscular junctions by visualizing horseradish peroxidase uptake in recycled synaptic vesicles. Direct innervation of the accessory radula closer muscle by B15 and B16 is demonstrated by experiments in which horseradish peroxidase is transported from motoneuronal soma to the terminals on muscle fibers. In addition, specific synaptic contacts between B4 and B5 and each of the motoneurons are observed in the buccal ganglion neuropil. Finally, multiple contacts consistent with peptidergic, serotoninergic, and cholinergic synapses are made onto the neurons, suggesting that a variety of transmitters modulate motor output at each level of the hierarchical circuit. These results support the physiological evidence suggesting the involvement of neuropeptides as well as "classical" transmitters in the modulation of circuitry governing feeding behavior in Aplysia.

Highly active neuromuscular system in developing lobsters with programmed obsolescence.

The primary locomotory apparatus in the three larval stages of the lobster, Homarus americanus, are paddlelike structures on the thoracic appendages called exopodites, which beat almost continuously. Consequently their power and return-stroke muscles are examples of highly active but short-lived neuromuscular systems. The muscles, which are well vascularized, are of the fast type with 2-3-micron sarcomere lengths and 6 thin filaments surrounding a thick one. The most striking feature, however, is the large volume of mitochondria making up 40-50% of the fiber. They appear as simple cylinders packed several layers deep along the periphery of the fiber and as large, multibranched forms distributed throughout the fiber and subdividing it into smaller units. The motor innervation to the return-stroke muscle is via 3 excitatory axons, which generate large junctional potentials and twitch contractions. The muscle is densely populated with large neuromuscular synapses, most of which have a well-defined active site or dense bar denoting the site of transmitter release. Altogether this motor system is specialized for prolonged activity. Atrophy of the neuromuscular system occurs by the late larval third stage. The muscle fibers lose their identity, fuse, and become vacuolated. The myofibrils condense and erode and the mitochondria are lost. Atrophy of motor innervation is gradual with individual axons dropping out. The largest axon providing most of the innervation is the first to degenerate. Early degenerative changes affect the axon and neuromuscular terminals but not the synaptic contacts, dense bars, and vesicles, which appear intact. Continued atrophy in the postlarval fourth stage reduces the exopodites to vestiges. Thus the return-stroke muscle of the larval exopodites in which muscle fiber and motoneurons are identifiable permits study of the interaction between a neuron and its target muscle undergoing programmed obsolescence.

Functional degeneration of isolated central stumps of crayfish sensory axons.

In the crayfish, Procambarus clarkii, nerve 5 carries primarily sensory axons from the tail fan to the 6th abdominal ganglion where they synaptically activate interneuron A. Since the sensory neurons have their somata located at the periphery, transection of nerve 5 part way to the ganglion allowed us to examine the fate of their soma-less central stumps. Up to 3 weeks postlesion the response to stimulation of nerve 5 consisted of a brief latency spike in interneuron A, similar to that in control animals and to stimulation of the intact nerve 4. Stimulation of the lesioned nerve 5 beyond 3 weeks failed to fire interneuron A. This loss of function was correlated to loss of axons in nerve 5 deduced by comparing the numbers in the lesioned nerve 5 to its contralateral intact counterpart. The numbers are about equal in the paired nerves but rapidly decline on the lesioned side to 50% within 1 week, 20% within 3 weeks, and less than 10% in subsequent weeks. This loss affects all size classes of axons. However, in the 3 week lesioned nerve large glial infoldings subdivided some of the larger axons and single nuclei were seen in a few of the medium-sized axons. Possibly subdivision of large axons by glial infolding may introduce glial nuclei into axons.