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Use and abuse of potential rates in soil microbiology

Potential rate assays are used in soil microbial ecology to determine the rates of a functional process in environmental samples under a defined set of conditions. While they can be used appropriately to provide mechanistic insights, potential rates are also often used to estimate the abundance of specific taxonomic groups and their in situ activity. These estimates incorrectly assume that all contributing organisms in a community are active at a maximum rate under one set of optimal incubation conditions and that potential rates reflect activity in the soil. While investigators now recognise that populations within communities are physiologically diverse, they often ignore the consequent suboptimal activity, or even inactivity, of the majority of community members performing that function. In this short perspective article, we discuss when potential assays can be informative and highlight the underlying conceptual problems under circumstances where potential assays are misused, using potential nitrification rate (PNR) as an example. PNR was originally developed to estimate the size of active ammonia oxidising communities in environmental samples. It is routinely determined in short-term shaken slurry incubations by measuring assumed maximum rates of nitrate or nitrite production under optimal, non-substrate-limiting conditions. As with other functional processes, it is now recognised that a broad diversity of organisms contribute to aerobic ammonia oxidation in terrestrial and other habitats, and this diversity represents a substantial range of physiologies, including variation in substrate affinity, ammonia tolerance, cell specific activity and substrate preference. Despite this, PNR, and other potential rate assays, are often inappropriately used in an attempt to determine an ecologically relevant measurement of activity in soil. As with any potential assay, PNR has inherent biases towards particular functional groups and its use in investigating the ecology of ammonia oxidisers in natural systems should be carefully considered.

microbiology↗

Assessment of the use and quick preparation of saliva for rapid microbiological diagnosis of COVID-19.

The objective of this study was to assess the performance of direct real time RT-PCR detection of SARS-CoV-2 in heated saliva samples, avoiding the RNA isolation step. Oropharyngeal and nasopharyngeal swabs together with saliva samples were obtained from 51 patients clinically diagnosed as potentially having COVID-19. Two different methods were compared: 1. RNA was extracted from 500 l of sample using a MagNA Pure Compact Instrument with an elution volume of 50l and 2. 700{micro}L of saliva were heat-inactivated at 96{degrees}C for 15 minutes, and directly subjected to RT-PCR. One step real time RT-PCR was performed using 5 l of extracted RNA or directly from 5 l of heated sample. RT-PCR was performed targeting the SARS-CoV-2 envelope (E) gene region. Diagnostic performance was assessed using the results of the RT-PCR from nasopharyngeal and oropharyngeal swabs as the gold standard. The overall sensitivity, specificity, positive and negative predictive values were 81.08%, 92.86%, 96.77% and 65.00%, respectively when RNA extraction was included in the protocol with saliva, whereas sensitivity, specificity, positive and negative predictive values were 83.78%, 92.86%, 68.42% and 96.88%, respectively, for the heat-inactivation protocol. However, when the analysis was performed exclusively on saliva samples with a limited time from the onset of symptoms (<9 days, N=28), these values were 90%, 87.5%, 44% and 98.75% for the heat-inactivation protocol. The study showed that RT-PCR can be performed using saliva in an RNA extraction free protocol, showing good sensitivity and specificity.

microbiology↗

Modeling by artificial neural networks of silver carp (Hypophthalmichthys molitrixi) with sous vide processing on the effects of storage and processing temperatures on the microbiological status

To evaluate and anticipate the microbial changes of silver carp (Hypophthalmichthys molitrixi) during cold storage (0, 5, 10, 15 & 21 day) at different sous vide processing temperatures (60, 65, 70, and 75 {degrees}C), changes in microbial load of Enterobacteriaceae, Lactic Acid bacteria (LAB), Pseudomonas, Psychrotrophs, and total viable count (TVC) were considered. A radial basis function neural network (RBFNN) model was established to predict the changes in the microbial content of silver carp. The critical temperature for inactivation of Enterobacteriaceae and lactic acid bacteria was 65 {degrees}C and for Pseudomonas and Psychrotrophs was 70 {degrees}C and the highest value (75 {degrees}C) was observed for the total viable count. In samples processed at 75 {degrees}C, the presence of Enterobacteriaceae, Pseudomonas and Psychrotrophs were not detectable up to 15 days of storage and lactic acid bacteria were not detectable even at the end of the storage period. The optimal ANN topology for modeling Enterobacteriaceae, Pseudomonas, and Psychrotroph contained 9 neurons in the hidden layer, but for TVC and LAB, it was 14 neurons.

microbiology↗

Spatial-temporal targeted and non-targeted surveys to assess microbiological composition of drinking water in Puerto Rico following Hurricane Maria.

Loss of basic utilities, such as drinking water and electricity distribution, were sustained for months in the aftermath of Hurricane Marias (HM) landfall in Puerto Rico (PR) in September 2017. The goal of this study was to assess if there was deterioration in biological quality of drinking water due to these disruptions. This study characterized the microbial composition of drinking water following HM across nine drinking water systems (DWSs) in PR and utilized an extended temporal sampling campaign to determine if changes in the drinking water microbiome were indicative of HM associated disturbance followed by recovery. In addition to monitoring water chemistry, the samples were subjected to culture independent targeted and non-targeted microbial analysis including quantitative PCR (qPCR) and genome-resolved metagenomics. The qPCR results showed that residual disinfectant was the major driver of bacterial concentrations in tap water with marked decrease in concentrations from early to late sampling timepoints. While Mycobacterium avium and Pseudomonas aeruginosa were not detected in any sampling locations and timepoints, genetic material from Leptospira and Legionella pneumophila were transiently detected in a few sampling locations. The majority of metagenome assembled genomes (MAGs) recovered from these samples were not associated with pathogens and were consistent with bacterial community members routinely detected in DWSs. Further, whole metagenome-level comparisons between drinking water samples collected in this study with samples from other full-scale DWS indicated no significant deviation from expected community membership of the drinking water microbiome. Overall, our results suggest that disruptions due to HM did not result in significant and sustained deterioration of biological quality of drinking water at our study sites. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=86 SRC="FIGDIR/small/442998v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@100c1e9org.highwire.dtl.DTLVardef@bae402org.highwire.dtl.DTLVardef@1cf41bborg.highwire.dtl.DTLVardef@1e46105_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

Expanding Clinical Phage Microbiology:‎Simulating Phage Inhalation for Respiratory Tract Infections ‎

Phage therapy is a promising antibacterial strategy for resistant respiratory tract infections. Phage inhalation may serve this goal; however, it requires a careful assessment of their delivery by this approach. Here we present an in-vitro model to evaluate phage inhalation. Eight phages, most of which target CF-common pathogens, were aerosolized and administered to a real-scale CT{square}derived 3D airways model with a breathing simulator. Viable phage loads reaching the output of the nebulizer and the tracheal level of the model were determined and compared to the loaded amount. Phage inhalation resulted in a diverse range of titer reduction, primarily associated with the nebulization process. No correlation was found between phage delivery to the phage physical or genomic dimensions. These findings highlight the need for tailored simulations of phage delivery, ideally by a patient-specific model in addition to proper phage matching, to increase the potential of phage therapy success. Take-Home MessagePhage therapy can be used against infectious diseases if personally tailored. Using a 3D airways model, we show that phage delivery by inhalation to the respiratory tract is unpredictable and also requires a precise evaluation.

microbiology↗

Microbiological quality assessment of five common foods sold at different points of sale in Burkina-Faso

The aim of the present study was to assess the microbial quality of five different types of food such as bread, pasta, rice with sauce, beans and milk sold in five localities of Burkina Faso. One hundred and one (101) samples were collected and microbial quality were assessed by evaluating the food hygiene indicators such as total aerobic mesophilic flora, total coliforms, thermotolerant coliforms, yeast and mould. Food safety indicators such as Escherichia coli, Salmonella, coagulase-positive staphylococci, Clostridium perfringens and Bacillus cereus were checked too. All samples were analyzed under ISO methods. The results showed that 73.27% of samples were satisfactory while 14.85% were acceptable and 11.88% were not satisfactory according to international standards. Among the food safety indicators sought, Escherichia coli was detected in two samples and Bacillus cereus in four samples. Most of the analyzed food exhibited good hygiene behavior within the acceptable limits and the highest of not satisfactory rate was observed in milk powder and rice with sauce. Ouagadougou samples record the highest number of not satisfactory samples. Despite the general quality was satisfactory, the presence of specific microorganisms such as coliforms is indicative of the poor hygiene surrounded these foods. It is therefore necessary to train and follow up the vendors in the handling of equipment, hand-washing practices and selling environment hygiene for better improvement of the quality of the street foods.

microbiology↗

A new microbiological weapon against lepidopteran pests

Nowadays researchers provide more and more evidence that it is necessary to develop an ecologically friendly approach to pest control. This is reflected in a sharp increase in the value of the biological insecticide market in recent decades. In our study, we found a virus strain belonging to the genus Cypovirus (Reoviridae); the strain was isolated from Dendrolimus sibiricus: that possesses attractive features as a candidate for mass production of biological agents for lepidopteran-pest control. We describe morphological, molecular, and ecological features of the new Cypovirus strain. This strain was found to be highly virulent to D. sibiricus (half-lethal dose is 68 occlusion bodies per second-instar larva) and to have a relatively wide host range (infects representatives of five families of Lepidoptera: Erebidae, Sphingidae, Pieridae, Noctuidae, and Lasiocampidae). The virus strain showed a strong interaction with a nontoxic adjuvant (optical brightener), which decreased the lethal dose for both main and alternative hosts, decreased lethal time, and may expand the host range. Moreover, we demonstrated that the insecticidal features were preserved after passaging through the most economically suitable host. By providing strong arguments for possible usefulness of this strain in pest control, we call on virologists, pest control specialists and molecular biologists to give more attention to the Cypovirus genus, which may lead to new insights in the field of pest control research and may provide significant advantages to compare with baculoviruses and Bacillus thuringiensis products which are nowadays main source of bioinsecticides. Significance statementWithin this article we are describing unique set of features of newly discovered cypovirus strain which possess by significant premises for modern biological insecticides requirements: high potency, universality, true regulating effect, flexible production (possibility to choose host species for production), interaction with enhancing adjuvants, ecologically friendly. Basing on genome alignment we suggest that increasing of host range of new strain is the sequence of evolutionary event which was occurred after coinfection of different CPV species within same host. This finding open new perspective to consider CPVs as perspective agent of biocontrol products.

microbiology↗

Eco-Microbiology: Discovering Biochemical Enhancers of PET Biodegradation by Piscinibacter sakaiensis

The scale of plastic pollution boggles the mind. Nearly 400 megatons of virgin plastics are produced annually, with an environmental release rate of 80 percent; and plastic waste including micro- and nanoplastics are associated with a plethora of problems. The naturally evolved abilities of plastic-degrading and consuming microbes offer a starting point for generating sustainable and eco-centric solutions to plastic pollution. Here we developed an iterative discovery procedure coupling faster quasi-high-throughput polyethylene terephthalate (PET) dependent bioactivity screens with longer-term PET biodegradation assays to find small molecule and ionic boosters of PET consumption by the bacterium Piscinibacter sakaiensis. We discovered multiple hits supporting greater than 2-fold enhancement of PET biodegradation - with hits belonging to a small but heterogeneous set of compounds and mixtures, suggesting upregulation of PET consumption via multiple paths. This work has the potential to advance the creation of a fermentation-based process for solving PET plastic pollution. ImportancePlastic pollution is an urgent environmental issue. In addition, micro- and nanoplastics (MNPs) have become an acute source of worry with discoveries of the global distribution and transport of MNPs, their presence within a diversity of organisms including common foodstuffs and human tissues, and their potential association with declining fertility and various disease states. Solutions are needed and the microbial world offers abundant help via naturally evolved biodegraders of plastic waste. We created a non-genetic method to accelerate polyethylene terephthalate (PET) plastic biodegradation by Piscinibacter sakaiensis, a bacterium that evolved to slowly but completely consume PET. Our method entails a combination of plastic-dependent bioactivity screens and slower biodegradation tests to find extrinsic biochemical stimulators of PET biodegradation. The conditions we found boost PET biodegradation by over two-fold and provide a foundation for further studies to realize a fermentation-based process needed to solve PET plastic pollution.

microbiology↗

Identifying the Fusarium species involved in foot rot disease of faba beans in the UK using a combined molecular and microbiological approach

Foot rot is a devastating disease of faba bean crops globally, including in the United Kingdom, the worlds third largest producer. To identify the causal agents, we have sampled foot rot-affected plants and soils from faba bean crops across England. We isolated organisms associated with foot rot disease in culture and assessed pathogenicity in vivo to evaluate the infectivity of the isolates on faba bean. We identified the pathogenic isolates using DNA barcoding of the Internal Transcribed Spacer (ITS) and Translation Elongation Factor one (TEF1 ) molecular markers. A total of 113 clonal isolates were obtained from infected plants and soil samples across England. Of these, 60 were pathogenic, inducing mild to severe symptoms on faba bean. Sequencing of the ITS and TEF1 loci and comparison against sequence databases (Genbank and Fusarium_ID) enabled the identification of pathogenic isolates, in decreasing order of frequency, as Fusarium oxysporum (26.6 %), F. vanettenii (25%), F. redolens (15 %), F. solani (11.6%), F. culmorum (8.3 %), F. avenaceum (6.7 %), F. equiseti (1.7 %), F. clavum (1.7 %), Clonostachys rosea (1.7%) and Alternaria alternata (1.7%). F. oxysporum, F. redolens and F. avenaceum induced the most severe symptoms, whilst F. solani induced the least severe symptoms. Determining the most prevalent causal agents of foot rot in UK faba beans will facilitate targeted disease monitoring and intervention for enhanced productivity.

microbiology↗

Rapid microbiology instrumentation for high-accuracy quantitative water safety assessments in remote and low-resource locations

Microbial contamination of surface waters remains a significant public health concern, particularly in low- and middle-income countries (LMICs) and urban settings affected by combined sewer overflows (CSOs). Standard culture-based methods for monitoring fecal indicator bacteria (FIB)-- including membrane filtration (MF) and most probable number (MPN) assays--typically require laboratory infrastructure, do not provide same-day results, and systematically underestimate total FIB loads due to their inability to accurately quantify aggregate-bound bacteria. This study evaluates ALERT, an automated rapid method for culturable FIB quantification, as a viable field-deployable alternative to conventional laboratory techniques. ALERT employs a whole-sample approach that comprehensively captures both planktonic and aggregate-bound E. coli, thereby addressing critical limitations of traditional methods. Three instrument variants were assessed: the in-situ ALERT System V2, the portable ALERT LAB, and the handheld ALERT One, designed for single-sample analysis in resource-limited environments. Comparative field and laboratory trials conducted in Germany, France, and the UK demonstrate that ALERT delivers accuracy and precision equivalent to MPN methods, while providing faster results directly in the field and enabling high-frequency sampling. Use cases included storm event monitoring, CSO impact evaluation, and on-site validation and optimization of point-of-use water potabilization treatments. ALERT One was also successfully used by citizen scientist volunteers, highlighting its suitability for decentralized community-based monitoring. By enabling rapid, accurate, and comprehensive microbial water quality assessments, ALERT qualifies as a robust field alternative to laboratory testing. It supports enhanced water safety, empowers community-based surveillance and public engagement, and facilitates data-driven decision-making in both LMICs and high-income countries.

microbiology↗

SimUrine: A Novel, Fully Defined Artificial Urinary Media for Enhanced Microbiological Research of Urinary Bacteria

Urinary tract infections represent one of the most prevalent bacterial diseases, yet current diagnostic and research methodologies are hampered by inadequate culture media that fail to replicate the bladder biochemical environment. Conventional artificial urine formulations contain undefined components, lack essential nutrients, or inadequately support urinary microbiome (urobiome) growth. To address these limitations, we developed SimUrine, a fully defined synthetic urine medium that aims to replicate human bladder chemistry while supporting diverse microbial growth requirements. SimUrine was systematically developed through iterative optimization of multi-purpose artificial urine, incorporating defined concentrations of carbon sources, vitamins, trace elements, and amino acids within physiologically relevant ranges. The modular design enables component substitution without complete reformulation, facilitating customization for culturomics, antimicrobial susceptibility testing, and microbial ecology studies, while reducing batch-to-batch variability associated with authentic urine. Performance evaluation demonstrated SimUrines capability to support growth of fastidious urobiome members, including Lactobacillus species, Aerococcus urinae, and Corynebacterium riegelii, which fail to proliferate in conventional minimal media. Physicochemical characterization confirmed that SimUrine formulation exhibits properties within normal human urine ranges for density, conductivity, osmolarity, and viscosity, ensuring physiological relevance. Clinical applications revealed reduced antibiotic susceptibility compared to standard media, suggesting more accurate representation of in vivo conditions. Co-culture experiments using Escherichia coli and Enterococcus faecalis demonstrated previously unobserved microbial interactions, highlighting SimUrines utility for investigating urobiome dynamics. SimUrine represents a significant advancement in urobiome research methodology, providing a standardized, reproducible platform for investigating urobiome under physiologically relevant conditions, potentially improving fundamental understanding and clinical diagnostic approaches. IMPORTANCEUrinary tract infections affect millions globally, yet current research and diagnostic methods rely on inadequate culture media that fail to replicate the bladders unique biochemical environment. This fundamental limitation has hindered accurate UTI research and potentially compromised clinical treatment decisions. SimUrine addresses this critical gap as the first fully defined synthetic urine medium that mimics human bladder chemistry while supporting growth of diverse urinary microbes. The breakthrough enables cultivation of urobiome organisms in a minimal medium that resembles natural conditions, revealing novel microbial interactions that influence urinary health. Crucially, SimUrine demonstrates different antimicrobial susceptibility patterns compared to standard clinical media, suggesting current testing protocols may inaccurately predict treatment outcomes. This standardized, reproducible platform eliminates the variability of authentic urine samples while maintaining physiological relevance, potentially transforming urobiome research methodology and improving clinical diagnostic accuracy for urinary tract infections worldwide.

microbiology↗

The microbiology and geochemistry of the shallow-water hydrothermal vents of the Gulf of Naples, Italy

Shallow-water hydrothermal vents are dynamic ecosystems that occur below 200 m in tectonically active regions of the planet. While their geochemical composition has been investigated in several locations, knowledge about the microbial diversity they harbour remains scarce. Moreover, the relationships between hydrothermal fluid chemistry, geological settings and microbial community structure in shallow vents have not been explored in detail. Here, we investigate the interplay between fluid geochemistry and microbial diversity in two underwater volcanic regions in the Gulf of Naples, Italy, one under the influence of the Somma-Vesuvio volcano and the other located within the underwater portion of the Campi Flegrei caldera. By combining 16S rRNA amplicon sequencing with geochemical measurements, and by contextualizing it with previous geochemical measurements done in the region, we found that hydrothermal fluid chemistry, influenced by the geological setting where the vents are hosted, plays a key role in shaping microbial ecological niches, and imposes strong selective pressures on the resident microbial communities. We additionally describe two new shallow vent sites, contributing to the characterization of the hydrothermalism in the area and unveiling the biodiversity associated with shallow-water hydrothermalism in the region.

microbiology↗

Epidemiology, Microbiology and Therapeutic Consequences of Chronic Osteomyelitis in Northern China: A Retrospective Analysis of 255 Patients

The study aimed to explore the epidemiology and clinical characteristics of chronic osteomyelitis observed in a northern China hospital. Clinical data of 255 patients with chronic osteomyelitis from January 2007 to January 2014 were collected and analyzed, including general information, disease data, treatment and follow-up data. Chronic osteomyelitis is more common in males and in the age group from 41-50 years of age. Common infection sites are the femur, tibiofibular, and hip joint. More g+ than g- bacterial infections were observed, with S. aureus the most commonly observed pathogenic organism. The positive detection rate from debridement bacterial culture is 75.6%. The detection rate when five samples are sent for bacterial culture is 90.6%, with pathogenic bacteria identified in 82.8% of cases. The two-stage debridement method (87.0%) has higher first curative rate than the one-stage debridement method (71.2%). To improve detection rate using bacterial culture, at least five samples are recommended. Treatment of chronic osteomyelitis with two-stage debridement, plus antibiotic-loaded polymethylmethacrylate (PMMA) beads provided good clinical results in this study and is therefore recommended.

microbiology↗

Physicochemical and microbiological profile of the potable water of Eastern Himalayan State Sikkim: An Indication of Severe Fecal Contamination and Immediate Health Risk

Increasing population, rapid urbanization and climate change have immensely affected the freshwater sources around the world. The continuous decline in the number of natural potable water sources raises serious concern about the overall health of the human population. Developing countries are the most affected in this regard due to lack of proper hygienic maintenance protocols. Sikkim an Eastern Himalayan state with mountains as predominant topological features, harbors several natural spring water (SW). These spring waters are the primary source of potable water for the population in the four districts of the state viz. East (E), West (W), South (S) and North (N). Several incidences of water-borne diseases and the relative lack of scientific evaluation reports on the water quality of the area have educed this study. Lack of any standard filtration and purification practice among the population is one of the prime factors for the outspread of different waterborne pathogen in the state. The people of the state mostly use boiling as a dominant method of water purification, while only a small percentage of people in the West district were found to use modern standard purification system (W = 30%). The rainy season was found to be the major contributor of different diseases (E = 86%; W=100%; S=100%; N=80%) and statistical analysis of the fecal coliforms of the different season also indicated a significant difference at p < 0.05. There was no statistical significance among the physicochemical parameter of the SW but surprisingly the water from the four districts was recorded with traces of highly toxic heavy metals like mercury/WHO limit (0.001-0.007mg/l/0.001) as well as lead/WHO limit (0.001-0.007mg/l/0.05) and selenium/WHO limit (0.526-0.644 mg/l/0.01) which was above the WHO permissible limit. Piper analysis showed that water was dominated by cation sodium ion (Na+) and bicarbonate (HCO3-) anion and the water can be categorized as Mg-HCO3- type. Pairwise Pearson correlation showed a significant correlation between Electrical conductivity and TDS (r = 0.998/1.00) as well as alkalinity and turbidity (0.993/1.00). The microbial confirmatory test showed severity in fecal contamination with high counts of Total Coliform (TC), Escherichia coli (EC) and Enterococcus (EN). Highest TC was recorded from W (37.26/ml) and lowest in N (22.13/ ml) in spring water. Highest contamination of Escherichia coli and Enterococcus was found in E (EC = 8.7/ml; EN = 2.08/ml) followed by S (EC = 8.4/ml and EN = 2.05/ml). It was found that community reservoir (CR) tank was more contaminated than SP, which indicates the negligence in maintenance and fecal contamination during transportation to the reservoir. Though household water was least contaminated compared to CR and SP, but it fails in WHO standard criteria for drinking water. These results indicate an immediate health risk of the resident of the state and which needs to be taken care of sooner as possible by protecting the important potable sources with required policies and regulations.

microbiology↗

Ecological and microbiological diversity of chigger mites, including vectors of scrub typhus, on small mammals across stratified habitats in Thailand

Scrub typhus, caused by a bacterial pathogen (Orientia spp.), is a potentially life-threatening febrile illness widely distributed in the Asia-Pacific region and is emerging elsewhere. The infection is transmitted by the larval stage of trombiculid mites (\"chiggers\") that often exhibit low host specificity. Here, we present an analysis of chigger ecology for 38 species sampled from 11 provinces of Thailand and microbiomes for eight widespread species. In total, >16 000 individual chiggers were collected from 1 574 small mammal specimens belonging to 18 species across four horizontally-stratified habitat types. Chigger species richness was positively associated with higher latitudes, dry seasonal conditions, and host maturity; but negatively associated with increased human land use. Human scrub typhus incidence was found to be positively correlated with chigger species richness. The bacterial microbiome of chiggers was highly diverse, with Sphingobium, Mycobacterium, Neisseriaceae and various Bacillales representing the most abundant taxa. Only Leptotrombidium deliense was found to be infected with Orientia. {beta}-diversity, but not -diversity, was significantly different between chigger species and geographic regions, although not between habitat types. This first field survey of the chigger microbiome provides a framework for future studies on interactions between pathogens and other symbionts in these understudied vectors.

microbiology↗

Microbiological, biochemical, physicochemical surface properties and biofilm forming ability of Brettanomyces bruxellensis

Brettanomyces bruxellensis is a serious source of concern for winemakers. The production of volatile phenols by the yeast species confers to wine unpleasant sensory characteristics which are unacceptable by the consumers and inevitably provoke economic loss for the wine industry. This ubiquitous yeast is able to adapt to all winemaking steps and to withstand various environmental conditions. Moreover, the ability of B. bruxellensis to adhere and colonize inert materials can be the cause of the yeast persistence in the cellars and thus recurrent wine spoilage. We therefore investigated the surface properties, biofilm formation capacity and the factors which may affect the attachment of the yeast cells to surfaces with eight strains representative of the genetic diversity of the species. Our results show that the biofilm formation ability is strain-dependent and suggest a possible link between the physicochemical properties of the studied strains and their corresponding genetic group.

microbiology↗

SRST2: Rapid genomic surveillance for public health and hospital microbiology labs

Rapid molecular typing of bacterial pathogens is critical for public health epidemiology, surveillance and infection control, yet routine use of whole genome sequencing (WGS) for these purposes poses significant challenges. Here we present SRST2, a read mapping-based tool for fast and accurate detection of genes, alleles and multi-locus sequence types (MLST) from WGS data. Using >900 genomes from common pathogens, we show SRST2 is highly accurate and outperforms assembly-based methods in terms of both gene detection and allele assignment. Here we have demonstrated the use of SRST2 for microbial genome surveillance in a variety of public health and hospital settings. In the face of rising threats of antimicrobial resistance and emerging virulence amongst bacterial pathogens, SRST2 represents a powerful tool for rapidly extracting clinically useful information from raw WGS data. Source code is available from http://katholt.github.io/srst2/.

Genomics↗

Evidence-Based Design and Evaluation of a Whole Genome Sequencing Clinical Report for the Reference Microbiology Laboratory

BackgroundMicrobial genome sequencing is now being routinely used in many clinical and public health laboratories. Understanding how to report complex genomic test results to stakeholders who may have varying familiarity with genomics - including clinicians, laboratorians, epidemiologists, and researchers - is critical to the successful and sustainable implementation of this new technology; however, there are no evidence-based guidelines for designing such a report in the pathogen genomics domain. Here, we describe an iterative, human-centered approach to creating a report template for communicating tuberculosis (TB) genomic test results.\n\nMethodsWe used Design Study Methodology - a human centered multi-stage approach drawn from the information visualization domain - to redesign an existing clinical report. We used expert consults and an online questionnaire to discover various stakeholders needs around the types of data and tasks related to TB that they encounter in their daily workflow. We also evaluated their perceptions of and familiarity with genomic data, as well as its utility at various clinical decision points. These data shaped the design of multiple prototype reports that were compared against the existing report through a second online survey, with the resulting qualitative and quantitative data informing the final, redesigned, report.\n\nResultsWe recruited 78 participants, 65 of whom were clinicians, nurses, laboratorians, researchers, and epidemiologists involved in TB diagnosis, treatment, and/or surveillance. Our first survey indicated that participants were largely enthusiastic about genomic data, with the majority agreeing on its utility for certain TB diagnosis and treatment tasks and many reporting some confidence in their ability to interpret this type of data (between 58.8% and 94.1%, depending on the specific data type). When we compared our four prototype reports against the existing design, we found that for the majority (86.7%) of design comparisons, participants preferred the alternative prototype designs over the existing version, and that both clinicians and non-clinicians expressed similar design preferences. Participants articulated clearer design preferences when asked to compare individual design elements versus entire reports. Both the quantitative and qualitative data informed the design of a revised report, which is available online as a LaTeX template.\n\nConclusionsWe show how a human-centered design approach integrating quantitative and qualitative feedback can be used to design an alternative report for representing complex microbial genomic data. We suggest experimental and design guidelines to inform future design studies in the bioinformatics and microbial genomics domains, and suggest that this type of mixed-methods study is important to facilitate the successful translation of pathogen genomics in the clinic, not only for clinical reports but also more complex bioinformatics data visualization software.

genomics↗