Environmental Health

Legionella spp. are the causative agent of Legionnaire's disease and an opportunistic pathogen of significant public health concern. Identification and quantification from environmental sources is crucial for identifying outbreak origins and providing sufficient information for risk assessment and disease prevention. Currently there are a range of methods for Legionella spp. quantification from environmental sources, but the two most widely used and accepted are culture and real-time polymerase chain reaction (qPCR). This paper provides a review of these two methods and outlines their advantages and limitations. Studies from the last 10 years which have concurrently used culture and qPCR to quantify Legionella spp. from environmental sources have been compiled. 26/28 studies detected Legionella at a higher rate using qPCR compared to culture, whilst only one study detected equivalent levels of Legionella spp. using both qPCR and culture. Aggregating the environmental samples from all 28 studies, 2856/3967 (72%) tested positive for the presence of Legionella spp. using qPCR and 1331/3967 (34%) using culture. The lack of correlation between methods highlights the need to develop an acceptable standardized method for quantification that is sufficient for risk assessment and management of this human pathogen.

Campylobacteriosis is infection caused by the bacteria Campylobacter spp. and is considered a major public health concern. Campylobacter spp. have been identified as one of the most common causative agents of bacterial gastroenteritis. They are typically considered a foodborne pathogen and have been shown to colonise the intestinal mucosa of all food-producing animals. Much emphasis has been placed on controlling the foodborne pathway of exposure, particularly within the poultry industry, however, other environmental sources have been identified as important contributors to human infection. This paper aims to review the current literature on the sources of human exposure to Campylobacter spp. and will cover contaminated poultry, red meat, unpasteurised milk, unwashed fruit and vegetables, compost, wild bird faeces, sewage, surface water, ground water and drinking water. A comparison of current Campylobacter spp. identification methods from environmental samples is also presented. The review of literature suggests that there are multiple and diverse sources for Campylobacter infection. Many environmental sources result in direct human exposure but also in contamination of the food processing industry. This review provides useful information for risk assessment.

This study used Fluorescent in situ Hybridisation (FISH) with rRNA targeted oligonucleotide probes combined with scanning confocal laser microscopy to successfully detect Legionella spp. in commercially available potting mix. A range of techniques were explored to optimise the FISH method by reducing background fluorescence and preventing non-specific binding of probes. These techniques included the use of a blocking agent, UV light treatment, image subtraction of a nonsense probe and spectral unmixing of specific probes fluorescence and autofluorescence dependent on the specific emission spectra of probe fluorophores. Spectral unmixing was the best microscopy technique for reducing background fluorescence and non-specific binding of probes was not observed. The rapid turnaround time and increased sensitivity of the FISH provides as an alternative to traditional culture methods, which are tedious and often give varied results. FISH is also advantageous compared to PCR methods as it provides information on the structure of the microbial community the bacteria is situated in. This study demonstrates that FISH could provide an alternative method for Legionella detection and enumeration in environmental samples. j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / j m i c m e t h Please cite this article as: Whiley, H., et al., Detection of Legionella species in potting mixes using fluorescent in situ hybridisation (FISH),

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.

Inhalation of potable water presents a potential route of exposure to opportunistic pathogens and hence warrants significant public health concern. This study used qPCR to detect opportunistic pathogens Legionella spp., L. pneumophila and MAC at multiple points along two potable water distribution pipelines. One used chlorine disinfection and the other chloramine disinfection. Samples were collected four times over the year to provide seasonal variation and the chlorine or chloramine residual was measured during collection. Legionella spp., L. pneumophila and MAC were detected in both distribution systems throughout the year and were all detected at a maximum concentration of 10 3 copies/mL in the chlorine disinfected system and 10 6 , 10 3 and 10 4 copies/mL respectively in the chloramine disinfected system. The concentrations of these opportunistic pathogens were primarily controlled throughout the distribution network through the maintenance of disinfection residuals. At a dead-end and when the disinfection residual was not maintained significant (p < 0.05) increases in concentration were observed when compared to the concentration measured closest to the processing plant in the same pipeline and sampling period. Total coliforms were not present in any water sample collected. This study demonstrates the ability of Legionella spp., L. pneumophila and MAC to survive the OPEN ACCESS potable water disinfection process and highlights the need for greater measures to control these organisms along the distribution pipeline and at point of use.

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.

Foodborne illness is a global public health burden. Over the past decade in Australia, despite advances in microbiological detection and control methods, there has been an increase in the incidence of foodborne illness. Therefore improvements in the regulation and implementation of food safety policy are crucial for protecting public health. In 2000, Australia established a national food safety regulatory system, which included the adoption of a mandatory set of food safety standards. These were in line with international standards and moved away from a &quot;command and control&quot; regulatory approach to an &quot;outcomes-based&quot; approach using risk assessment. The aim was to achieve national consistency and reduce foodborne illness without unnecessarily burdening businesses. Evidence demonstrates that a risk based approach provides better protection for consumers; however, sixteen years after the adoption of the new approach, the rates of food borne illness are still increas...

Legionella is an opportunistic pathogen of public health significance. One of the main sources of Legionella is potable water systems. As a consequence of aging populations there is an increasing demographic considered at high risk for Legionellosis and, as such, a review of the guidelines is required. Worldwide, Legionella has been detected from many potable water sources, suggesting it is ubiquitous in this environment. Previous studies have identified the limitations of the current standard method for Legionella detection and the high possibility of it returning both false negative and false positive results. There is also huge variability in Legionella test results for the same water sample when conducted at different laboratories. However, many guidelines still recommend the testing of water systems. This commentary argues for the removal of routine Legionella monitoring from all water distribution guidelines. This procedure is financially consuming and false negatives may resu...

Globally, there has been an increase in squamates (particularly lizards and snakes) being kept as pets. Additionally, urban spread has resulted in greater human encroachment and interaction with the natural habitat of wild squamates. A potential consequence of increasing human interaction with squamates is the increased potential for disease transfer. This review collates the literature describing clinical salmonellosis cases that were definitively linked to a squamate through testing of the animal and population-based studies which investigate the risk of salmonellosis linked to pet squamates. It was demonstrated that although squamate-acquired salmonellosis accounted for a small percentage of total cases, children under five were at greatest risk, with the clinical manifestations tending to be more severe. In many cases, it was noted that the patient was unaware of the risks associated with keeping squamates and did not practice proper hand hygiene after handling the animals or cleaning cages. This highlights the need for more education focused on informing the general public of ways to reduce the risk of salmonellosis from pet squamates. There is also the need for future research into the role of wild squamates in the spread of human salmonellosis, both directly and indirectly through cross contamination.

Quantitative microbial risk assessment (QMRA) is a relatively new approach in identifying health risks associated with the ubiquitous presence of pathogens and opportunists in the human environment. The methodology builds on experimental and meta-analytical data to identify measurable factors that contribute to, and can quantify, the likely extent of disease given a particular exposure. Early modelling was particularly focused on food-borne disease, and subsequently water-borne disease, with the emphasis focused on ingestion and its role in enteric disease. More recently, there has been a focus on translating these principles to opportunist waterborne infections (OWI) with primary focus on Legionella spp. Whereas dose and susceptibility are well documented via the ingestion route of exposure there is considerably less certainty regarding both factors when understanding Legionella spp. and other OWI. Many OWI can arise through numerous routes of transmission with greatly differing disease presentations. Routes of Legionella spp. infection do not include ingestion, but rather aspiration and inhalation of contaminated water are the routes of exposure. The susceptible population for OWI is a vulnerable subset of the population unlike those associated with enteric disease pathogens. These variabilities in dose, exposure and susceptibility call in to question whether QMRA can be a useful tool in managing risks associated with OWI. Consideration of Legionella spp. as a well-documented subject of research calls into question whether QMRA of OWI is likely to be a useful tool in developing risk management strategies.

Foodborne illness is a global public health issue, with food handling in the home identified as an underestimated source. In Australia, there has been a significant increase in the incidence of salmonellosis with the majority of outbreaks linked to eggs. This study investigated Australian eggs consumer attitudes, behaviours and risk perceptions associated with the handling of raw eggs in the home. It was identified that 67% of participants chose free range eggs, 11% kept poultry, 7% did not have any preference, 7% cage eggs, 4% barn eggs, 2% organic eggs and 1% pasteurized eggs. The majority of participants (91%) reported they stored eggs in the fridge. It was identified that there is an underestimation of &quot;risky behaviour&quot; associated with the consumption of raw eggs in the home, as 84% of participants indicated that they did not consume raw eggs, but subsequently 86% indicated that they had eaten mixture/batter containing raw eggs. Participants&#39; responses relating to ...

Globally, there has been an increase in squamates (particularly lizards and snakes) being kept as pets. Additionally, urban spread has resulted in greater human encroachment and interaction with the natural habitat of wild squamates. A potential consequence of increasing human interaction with squamates is the increased potential for disease transfer. This review collates the literature describing clinical salmonellosis cases that were definitively linked to a squamate through testing of the animal and population-based studies which investigate the risk of salmonellosis linked to pet squamates. It was demonstrated that although squamate-acquired salmonellosis accounted for a small percentage of total cases, children under five were at greatest risk, with the clinical manifestations tending to be more severe. In many cases, it was noted that the patient was unaware of the risks associated with keeping squamates and did not practice proper hand hygiene after handling the animals or cleaning cages. This highlights the need for more education focused on informing the general public of ways to reduce the risk of salmonellosis from pet squamates. There is also the need for future research into the role of wild squamates in the spread of human salmonellosis, both directly and indirectly through cross contamination.