The effect of temperature fluctuation is an important factor in bacterial growth especially for pathogens such as the staphylococci that have to remain viable during potentially harsh and prolonged transfer conditions between hosts. The aim of this study was to investigate the response of S. aureus, S. epidermidis, and S. lugdunensis when exposed to low temperature (4°C) for prolonged periods, and how this factor affected their subsequent growth, colony morphology, cellular ultra-structure, and amino acid composition in the non-cytoplasmic hydrolysate fraction. Clinical isolates were grown under optimal conditions and then subjected to 4°C conditions for a period of 8 wks. Cold-stressed and reference control samples were assessed under transmission electron microscopy (TEM) to identify potential ultra-structural changes. To determine changes in amino acid composition, cells were fractured to remove the lipid and cytoplasmic components and the remaining structural components were hydrolysed. Amino acid profiles for the hydrolysis fraction were then analysed for changes by using principal component analysis (PCA). Exposure of the three staphylococci to prolonged low temperature stress resulted in the formation of increasing proportions of small colony variant (SCV) phenotypes. TEM revealed that SCV cells had significantly thicker and more diffuse cell-walls than their corresponding WT samples for both S. aureus and S. epidermidis, but the changes were not significant for S. lugdunensis. Substantial species-specific alterations in the amino acid composition of the structural hydrolysate fraction were also observed in the cold-treated cells. The data indicated that the staphylococci responded over prolonged periods of cold-stress treatment by transforming into SCV populations. The observed ultra-structural and amino acid changes were proposed to represent response mechanisms for staphylococcal survival amidst hostile conditions, thus maintaining the viability of the species until favourable conditions arise again.

This study investigated whether alterations in environmental conditions would induce the formation of small colony variant phenotypes (SCV) with associated changes in cell morphology and ultra-structure in S. aureus, s. epidermidis, and S. lugdunensis. Wild-type clinical isolates were exposed to low temperature (4°C), antibiotic stress (penicillin G and vancomycin; 0-10,000 µg mL-1), pH stress (pH 3-9) and osmotic challenge (NaCl concentrations of 0-20%). Changes in cell diameter, cell-wall thickness, and population distribution changes (n ≥ 300) were assessed via scanning and transmission electron microscopy (SEM and TEM), and compared to control populations. Our analyses found that prolonged exposure to all treatments resulted in the subsequent formation of SCV phenotypes. Observed SCVs manifested as minute colonies with reduced haemolysis and pigmentation (NaCl, pH and 4°C treatments), or complete lack thereof (antibiotic treatments). SEM comparison analyses revealed significantly smaller cell sizes for SCV populations except in S. aureus and S. epidermidis 10% NaCl, and S. epidermidis 4°C (p<0.05). Shifts in population distribution patterns were also observed with distinct sub-populations of smaller cells appearing for S. epidermidis, and S. lugdunensis. TEM analyses revealed significantly thicker cell-walls in all treatments and species except S. lugdunensis exposed to 4°C. These findings suggest that staphylococci adapted to environmental stresses by altering their cell size and wall thickness which could represent the formation of altered phenotypes which facilitate survival under harsh conditions. The phenotypic response was governed by the type of prevailing environmental stress regime leading to appropriate alterations in ultra-structure and size, suggesting downstream changes in gene expression, the proteome, and metabolome.

Staphylococci are highly successful at colonizing a variety of dynamic environments, both nonpathogenic and those of clinical importance, and comprise the list of pathogens of global public health significance. Their remarkable survival and persistence can be attributed to a host of strategies, one of which is metabolic versatility—their ability to rapidly alter their metabolism in the presence of transient or long-term bacteriostatic and bactericidal conditions and facilitate cellular homeostasis. These attributes contribute to their widespread dissemination and challenging eradication particularly from clinical settings. The study of microbial behaviour at the metabolite level provides insight into mechanisms of survival and persistence under defined environmental and clinical conditions. This paper reviews the range of metabolic modulations that facilitate staphylococcal acclimatization and persistence in varying terrestrial and host conditions, and their public health ramifications in these settings.

BackgroundPassage of bacterial cells through filter pores has been reported for a number of bacterial species. In this investigation, we tested the filterability of staphylococcal cultures that were exposed to several environmental stress conditions by passing them through 0.22 and 0.45 μm sterile filters, which are industry standards.FindingsResults showed repeated passage of viable staphylococcal cells through both pore sizes, although more passage was seen through the 0.45 μm pore size. Of the three staphylococcal species, S. lugdunensis showed the best passage at relatively higher numbers regardless of the treatment, while both S. aureus and S. epidermidis showed limited passage or complete inhibition.ConclusionThe data showed that staphylococcal bacteria were capable of passing through sterile filters in a viable state. There was better passage through 0.45 μm sterile filters than through the 0.22 μm sterile filters. Application of a stress condition did not appear to enhance filterability of these bacterial cultures.

Potable reuse is implemented in several countries around the world to augment strained water supplies. This article presents a public health perspective on potable reuse by comparing the critical infrastructure and institutional capacity characteristics of two well-established potable reuse schemes with conventional drinking water schemes in developed nations that have experienced waterborne outbreaks. Analysis of failure events in conventional water systems between 2003 and 2013 showed that despite advances in water treatment technologies, drinking water outbreaks caused by microbial contamination were still frequent in developed countries and can be attributed to failures in infrastructure or institutional practices. Numerous institutional failures linked to ineffective treatment protocols, poor operational practices, and negligence were detected. In contrast, potable reuse schemes that use multiple barriers, online instrumentation, and operational measures were found to address the events that have resulted in waterborne outbreaks in conventional systems in the past decade. Syndromic surveillance has emerged as a tool in outbreak detection and was useful in detecting some outbreaks; increases in emergency department visits and GP consultations being the most common data source, suggesting potential for an increasing role in public health surveillance of waterborne outbreaks. These results highlight desirable characteristics of potable reuse schemes from a public health perspective with potential for guiding policy on surveillance activities.