Kirstin Ross

Legionella, Protozoa, and Biofilms: Interactions Within Complex Microbial Systems

Currently, the investigation of Legionella ecology falls into two distinct areas of research activity: (1) that Legionella multiply within water sources by parasitizing amoebic or ciliate hosts or (2) that Legionella grows extracellularly within biofilms. Less focus has been given to the overlaps that may occur between these two areas or the likelihood that Legionella employs multiple survival strategies to persist in water sources. It is likely that Legionella interacts with protozoa, bacteria, algae, fungi, etc., and biofilm components in a more complex fashion than multiplication or death due to the presence or absence of single components of these complex microbial systems. This paper addresses gaps that exist in the understanding of Legionella ecology and serves to pinpoint areas of future research. To assume that only one other class of organism is important to Legionella ecology may limit our understanding of how this bacterium proliferates in heated water sources and also limit our strategies for its control in the built environment.

Mercury in seafood: mechanisms of accumulation and consequences for consumer health.

Mercury is a largely uncontrollable heavy metal contaminant in that it is globally ubiquitous, and environmentally persistent. The element has the potential for global mobilization following liberation from environmental stores, which can occur as a consequence of either anthropogenic activities or natural processes. Furthermore, organic forms like methylmercury accumulate in biological tissues with an exceptionally long biological half-life, facilitating the magnification of this toxin along trophic food chains. Bioaccumulation is particularly evident in aquatic environments, in which long-lived piscivorous fishes and marine mammals are reported with a mercury burden one-million times that of the surrounding water body, typically attaining mercury burdens exceeding 1 microg g(-1). Mercury levels in other seafood, however, are typically reported in the range of 0.1 to 0.2 microg g(-1) and usually less then 0.5 microg g(-1). The primary source of human exposure to environmental mercury is through seafood consumption. The dangers associated with the consumption of large amounts of methylmercury accumulated in seafood are well recognized from past poisoning incidents, in which fish with mercury burdens in the range of 9 to 24 microg g(-1) were consumed. Nevertheless, the toxicological consequence of chronic low-level mercury exposure from habitual seafood consumption is an area of contention. This review discusses the mechanisms of mercury accumulation and distribution in fish tissues and the toxicological consequences of mercury exposure from seafood consumption with regard to international safety guidelines.

Conservation genetics and demographic history of the endangered Cape Fear shiner ( Notropis mekistocholas )

We examined allelic variation at 22 nuclear‐encoded markers (21 microsatellites and one anonymous locus) and mitochondrial (mt)DNA in two geographical samples of the endangered cyprinid fish Notropis mekistocholas (Cape Fear shiner). Genetic diversity was relatively high in comparison to other endangered vertebrates, and there was no evidence of small population effects despite the low abundance reported for the species. Significant heterogeneity (following Bonferroni correction) in allele distribution at three microsatellites and in haplotype distribution in mtDNA was detected between the two localities. This heterogeneity may be due to reduced gene flow caused by a dam built in the early 1900s. Bayesian coalescent analysis of microsatellite variation indicated that effective population size of Cape Fear shiners has declined in recent times (11–25 435 years ago, with highest posterior probabilities between 126 and 2007 years ago) by one–two orders of magnitude, consistent with the observed decline in abundance of the species. A decline in effective size was not indicated by analysis of mtDNA, where sequence polymorphism appeared to carry the signature of an older expansion phase that dated to the Pleistocene (∼12 700 > 1 million years ago). Cape Fear shiners thus appear to have undergone an expansion phase following a glacial cycle but to have declined significantly in more recent times. These results suggest that rapidly evolving markers such as microsatellites may constitute a suitable tool when inferring recent demographic dynamics of populations.

Salmonella and Eggs: From Production to Plate

Salmonella contamination of eggs and egg shells has been identified as a public health concern worldwide. A recent shift in consumer preferences has impacted on the egg industry, with a push for cage-free egg production methods. There has also been an increased desire from consumers for raw and unprocessed foods, potentially increasing the risk of salmonellosis. In response to these changes, this review explores the current literature regarding Salmonella contamination of eggs during the production processing through to food handling protocols. The contamination of eggs with Salmonella during the production process is a complex issue, influenced by many variables including flock size, flock age, stress, feed, vaccination, and cleaning routines. Currently there is no consensus regarding the impact of caged, barn and free range egg production has on Salmonella contamination of eggs. The literature regarding the management and control strategies post-collection, during storage, transport and food handling is also reviewed. Pasteurisation and irradiation were identified as the only certain methods for controlling Salmonella and are essential for the protection of high risk groups, whereas control of temperature and pH were identified as potential control methods to minimise the risk for foods containing raw eggs; however, further research is required to provide more detailed control protocols and education programs to reduce the risk of salmonellosis from egg consumption.

Limitations of Using Propidium Monoazide with qPCR to Discriminate between Live and Dead Legionella in Biofilm Samples

Accurately quantifying Legionella for regulatory purposes to protect public health is essential. Real-time PCR (qPCR) has been proposed as a better method for detecting and enumerating Legionella in samples than conventional culture method. However, since qPCR amplifies any target DNA in the sample, the techniques inability to discriminate between live and dead cells means that counts are generally significantly overestimated. Propidium monoazide (PMA) has been used successfully in qPCR to aid live/dead discrimination. We tested PMA use as a method to count only live Legionella cells in samples collected from a modified chemostat that generates environmentally comparable samples. Counts from PMA-treated samples that were pretreated with either heat or three types of disinfectants (to kill the cells) were highly variable, with the only consistent trend being the relationship between biofilm mass and numbers of Legionella cells. Two possibilities explain this result: 1. PMA treatment worked and the subsequent muted response of Legionella to disinfection treatment is a factor of biofilm/microbiological effects; although this does not account for the relationship between the amount of biofilm sampled and the viable Legionella count as determined by PMA-qPCR; or 2. PMA treatment did not work, and any measured decrease or increase in detectable Legionella is because of other factors affecting the method. This is the most likely explanation for our results, suggesting that higher concentrations of PMA might be needed to compensate for the presence of other compounds in an environmental sample or that lower amounts of biofilm need to be sampled. As PMA becomes increasingly toxic at higher concentrations and is very expensive, augmenting the method to include higher PMA concentrations is both counterproductive and cost prohibitive. Conversely, if smaller volumes of biofilm are used, the reproducibility of the method is reduced. Our results suggest that using PMA is not an appropriate method for discriminating between live and dead cells to enumerate Legionella for regulatory purposes.

Strongyloidiasis: A Disease of Socioeconomic Disadvantage.

Strongyloidiasis is a disease caused by soil transmitted helminths of the Strongyloides genus. Currently, it is predominately described as a neglected tropical disease. However, this description is misleading as it focuses on the geographical location of the disease and not the primary consideration, which is the socioeconomic conditions and poor infrastructure found within endemic regions. This classification may result in misdiagnosis and mistreatment by physicians, but more importantly, it influences how the disease is fundamentally viewed. Strongyloidiasis must be first and foremost considered as a disease of disadvantage, to ensure the correct strategies and control measures are used to prevent infection. Changing how strongyloidiasis is perceived from a geographic and clinical issue to an environmental health issue represents the first step in identifying appropriate long term control measures. This includes emphasis on environmental health controls, such as better infrastructure, sanitation and living conditions. This review explores the global prevalence of strongyloidiasis in relation to its presence in subtropical, tropical and temperate climate zones with mild and cold winters, but also explores the corresponding socioeconomic conditions of these regions. The evidence shows that strongyloidiasis is primarily determined by the socioeconomic status of the communities rather than geographic or climatic conditions. It demonstrates that strongyloidiasis should no longer be referred to as a “tropical” disease but rather a disease of disadvantage. This philosophical shift will promote the development of correct control strategies for preventing this disease of disadvantage.

Uncertainties associated with assessing the public health risk from Legionella

Legionella is an opportunistic pathogen of public health concern. Current regulatory and management guidelines for the control of this organism are informed by risk assessments. However, there are many unanswered questions and uncertainties regarding Legionella epidemiology, strain infectivity, infectious dose, and detection methods. This review follows the EnHealth Risk Assessment Framework, to examine the current information available regarding Legionella risk and discuss the uncertainties and assumptions. This review can be used as a tool for understanding the uncertainties associated with Legionella risk assessment. It also serves to highlight the areas of Legionella research that require future focus. Improvement of these uncertainties will provide information to enhance risk management practices for Legionella, potentially improving public health protection and reducing the economic costs by streamlining current management practices.

Empirical models to identify mechanisms driving reductions in tissue mercury concentration during culture of farmed southern bluefin tuna Thunnnus maccoyii

Two empirical models are presented to elucidate the mechanisms driving reductions in the mercury concentration of southern bluefin tuna (SBT) during culture. Model 1 predicts temporal fluctuations in mercury concentration in response to growth dilution. Model 2 predicts the combined effects of growth dilution and linear mercury accumulation. Model 2 was found to be the more accurate model. Over a typical farming period of 136 days, growth dilution resulted in a reduction in mean mercury concentration of SBT edible tissues from 0.51 mg/kg down to 0.33 mg/kg. Extended culture beyond 136 days resulted in an increase in mercury concentration due to the combined effects of mercury accumulation and seasonal lipid depletion. Results indicate that under current industry practice, cultured SBT can be consumed twice as frequently as that of wild caught SBT while maintaining total dietary mercury intake below national recommendations.

Human strongyloidiasis: identifying knowledge gaps, with emphasis on environmental control

Abstract Strongyloides is a human parasitic nematode that is poorly understood outside a clinical context. This article identifies gaps within the literature, with particular emphasis on gaps that are hindering environmental control of Strongyloides. The prevalence and distribution of Strongyloides is unclear. An estimate of 100–370 million people infected worldwide has been proposed; however, inaccuracy of diagnosis, unreliability of prevalence mapping, and the fact that strongyloidiasis remains a neglected disease suggest that the higher figure of more than 300 million cases is likely to be a more accurate estimate. The complexity of Strongyloides life cycle means that laboratory cultures cannot be maintained outside of a host. This currently limits the range of laboratory-based research, which is vital to controlling Strongyloides through environmental alteration or treatment. Successful clinical treatment with antihelminthic drugs has meant that controlling Strongyloides through environmental control, rather than clinical intervention, has been largely overlooked. These control measures may encompass alteration of the soil environment through physical means, such as desiccation or removal of nutrients, or through chemical or biological agents. Repeated antihelminthic treatment of individuals with recurrent strongyloidiasis has not been observed to result in the selection of resistant strains; however, this has not been explicitly demonstrated, and relying on such assumptions in the long-term may prove to be shortsighted. It is ultimately naive to assume that continued administration of antihelminthics will be without any negative long-term effects. In Australia, strongyloidiasis primarily affects Indigenous communities, including communities from arid central Australia. This suggests that the range of Strongyloides extends beyond the reported tropical/subtropical boundary. Localized conditions that might result in this extended boundary include accumulation of moisture within housing because of malfunctioning health hardware inside and outside the house and the presence of dog fecal matter inside or outside housing areas.

Principles Relevant to Health Research among Indigenous Communities.

Research within Indigenous communities has been criticised for lacking community engagement, for being exploitative, and for poorly explaining the processes of research. To address these concerns, and to ensure ‘best practice’, Jamieson, et al. (2012) recently published a summary of principles outlined by the NHMRC (2003) in “one short, accessible document”. Here we expand on Jamieson et al.’s paper, which while commendable, lacks emphasis on the contribution that communities themselves can make to the research process and how culturally appropriate engagement, can allow this contribution to be assured, specifically with respect to engagement with remote communities. Engagement started before the research proposal is put forward, and continued after the research is completed, has integrity. We emphasise the value of narratives, of understanding cultural and customary behaviours and leadership, the importance of cultural legitimacy, and of the need for time, not just to allow for delays, but to ensure genuine participatory engagement from all members of the community. We also challenge researchers to consider the outcomes of their research, on the basis that increasing clinical evidence does not always result in better outcomes for the community involved.