Variational Phylodynamic Inference Using Pandemic-scale Data

The ongoing global pandemic has sharply increased the amount of data available to researchers in epidemiology and public health. Unfortunately, few existing analysis tools are capable of exploiting all of the information contained in a pandemic-scale data set, resulting in missed opportunities for improved surveillance and contact tracing. In this paper, we develop the variational Bayesian skyline (VBSKY), a method for fitting Bayesian phylodynamic models to very large pathogen genetic data sets. By combining recent advances in phylodynamic modeling, scalable Bayesian inference and differentiable programming, along with a few tailored heuristics, VBSKY is capable of analyzing thousands of genomes in a few minutes, providing accurate estimates of epidemiologically relevant quantities such as the effective reproduction number and overall sampling effort through time. We illustrate the utility of our method by performing a rapid analysis of a large number of SARS-CoV-2 genomes, and demonstrate that the resulting estimates closely track those derived from alternative sources of public health data.     

Rapid Short-Read Sequencing and Aneuploidy Detection Using MinION Nanopore Technology

MinION is a memory stick–sized nanopore-based sequencer designed primarily for single-molecule sequencing of long DNA fragments (>6 kb). We developed a library preparation and data-analysis method to enable rapid real-time sequencing of short DNA fragments (<1 kb) that resulted in the sequencing of 500 reads in 3 min and 40,000–80,000 reads in 2–4 hr at a rate of 30 nt/sec. We then demonstrated the clinical applicability of this approach by performing successful aneuploidy detection in prenatal and miscarriage samples with sequencing in <4 hr. This method broadens the application of nanopore-based single-molecule sequencing and makes it a promising and versatile tool for rapid clinical and research applications.     

Inference of the Properties of the Recombination Process from Whole Bacterial Genomes

Patterns of linkage disequilibrium, homoplasy, and incompatibility are difficult to interpret because they depend on several factors, including the recombination process and the population structure. Here we introduce a novel model-based framework to infer recombination properties from such summary statistics in bacterial genomes. The underlying model is sequentially Markovian so that data can be simulated very efficiently, and we use approximate Bayesian computation techniques to infer parameters. As this does not require us to calculate the likelihood function, the model can be easily extended to investigate less probed aspects of recombination. In particular, we extend our model to account for the bias in the recombination process whereby closely related bacteria recombine more often with one another. We show that this model provides a good fit to a data set of Bacillus cereus genomes and estimate several recombination properties, including the rate of bias in recombination. All the methods described in this article are implemented in a software package that is freely available for download at http://code.google.com/p/clonalorigin/.     

The role of health care workers and antiviral drugs in the control of pandemic influenza

Until a vaccine against the new strain becomes available, the response to newly emerged pandemic influenza will consist of the use of antiviral drugs and measures that limit exposure to infectious individuals. These first-line defence measures include isolating cases upon diagnosis, reducing close contacts, the use of personal protective equipment and hygiene, and using antiviral drugs for treatment and prophylaxis. There are significant 'costs' associated with control measures, so to justify such interventions it is important to assess their potential to reduce transmission. In this paper, we determine the effect that a number of different antiviral interventions have on the reproduction number of infectives and the probability that an imported infection fades out, and determine parameter scenarios for which these interventions are able to eliminate an emerging pandemic of influenza. We also assess the role that health care workers play in transmission and the extent to which providing them with antiviral prophylaxis and personal protective equipment modifies this role. Our results indicate that this class requires protection to avoid a greatly disproportionate contribution to early infective numbers, and for the maintenance of a stable health care system. Further, we show that the role children play in increasing transmission is moderate, in spite of closer mixing with other children.     

NADH荧光法快速检测细菌总数

基于细菌胞内NADH的荧光特性及其在胞内含量稳定的特性, 建立一种快速检测细菌总数的新方法。该荧光法的NADH检测限为1 nmol/L, NADH含量在10 nmol/L~0.2 mmol/L间与荧光强度呈良好线性关系(R2 =0.9905)。经离心获得菌体细胞, 热Tris-HCl法提取胞内NADH, 以 342 nm为激发波长, 461 nm为发射波长测定提取液荧光强度, 1 h内可检测到样品1×104 CFU/mL菌数。结果表明该方法快速、灵敏、简便、重复性好, 可适用于食品卫生与安全、环境检测等领域活细菌数量的定量检测。     

Modeling antibiotic resistance in hospitals: the impact of minimizing treatment duration

Infections caused by antibiotic-resistant pathogens are a global public health problem. Numerous individual- and population-level factors contribute to the emergence and spread of these pathogens. An individual-based model (IBM), formulated as a system of stochastically determined events, was developed to describe the complexities of the transmission dynamics of antibiotic-resistant bacteria. To simplify the interpretation and application of the model's conclusions, a corresponding deterministic model was created, which describes the average behavior of the IBM over a large number of simulations. The integration of these two model systems provides a quantitative analysis of the emergence and spread of antibiotic-resistant bacteria, and demonstrates that early initiation of treatment and minimization of its duration mitigates antibiotic resistance epidemics in hospitals.     

Enhanced Staining of Bacterial Flagella Using Aged Mordant in the Silver Stain

Intensity of bacterial flagella staining using a modified silver stain was increased by aging the mordant for one week at room temperature. The use of aged mordant increased the apparent diameters of stained flagella and resulted in a darker stain. The mordant remained stable for at least four months at room temperature. The staining protocol presented allows application to liquid or solid cultures.     

The Saprotrophic Bacterial Complex in the Raised Peat Bogs of Western Siberia

The population density of bacteria in peat deposits along the landscape profile of the Vasyugan Marsh has been found to be as high as tens of millions of CFU/g peat. The abundance and diversity of bacteria increased with depth within the peat deposit, correlating with an increasing level of peat degradation. Variations in these parameters with depth and season were greater in peat deposits located in transaccumulative and transitional positions than in the sedge-sphagnum bogs located at the eluvial region of the profile. In the upper 1-m-thick layer of the peat deposits studied, bacilli, represented by five species, dominated, whereas, in the deeper layers, spirilla and myxobacteria prevailed. These bacteria are major degraders of plant polymers. Unlike the bacterial communities found in the peat deposits of European Russia, the dominant taxa in the studied peat deposits of western Siberia are represented by bacteria resistant to extreme conditions.__________Translated from Mikrobiologiya, Vol. 74, No. 4, 2005, pp. 545–551.Original Russian Text Copyright © 2005 by Golovchenko, Sannikova, Dobrovol’skaya, Zvyagintsev.     

Assessment of Bacterial Communities in Auriferous and Non-Auriferous Soils Using Genetic and Functional Fingerprinting

Understanding the microbial processes affecting the mobility of Au is important in the development of biogeochemical models describing the formation of secondary anomalies and Au grains in soils and deeper regolith materials. This study characterizes bacterial activity in auriferous soils that is linked to the microbially mediated solubilization of Au, as a result of production and consumption of free amino acids, which can form stable complexes with Au. Through the application of 16S rDNA fingerprinting and community level physiological profiling (CLPP), concurrently with Au mobility data, microcosm experiments have demonstrated the role that mobile Au plays in determining the structure and function of bacterial communities in auriferous soils. The bacterial community of auriferous soils displayed genetic differences compared to non-auriferous (background) soils associated with the appearance of Methylocella sp., Arthrobacter sp. and Bacillus sp., as well as functional differences in the utilization of D-Cellobiose, L-Serine, L-Phenylalanine, L-Arginine and N-Acetyl-D-Glucosamine. These results suggest that soil bacterial communities are linked to biogeochemical Au cycling, and that microbial fingerprinting analyses may be used as a screening tool in Au exploration to differentiate auriferous from background terrains.     

Enumeration of Active Cells in the Bacterioplankton of the Rybinsk Reservoir Using 5-Cyano-2,3-Ditolyl Tetrazolium Chloride: A Comparison with Other Methods

The enumeration of actively respiring bacterial cells in different biotopes of the littoral zone of the Rybinsk Reservoir during the spring period of ice thaw using the fluorescent dye 5-cyano-2,3-ditolyl tetrazolium chloride showed that bacterial communities growing on the bottom surface of the ice cover and in water overgrown by higher aquatic plants were most active. The number of active cells among individual bacterial cells averaged 20% and reached about 40% among aggregated and filamentous bacterial cells. The results of the count of active bacteria by this method were compared with those obtained by other methods.     

A Color Image Analysis Method for Assessment of Germination Based on Differential Fluorescence Staining of Bacterial Spores and Vegetative Cells Using Acridine Orange

Color fluorescence image analysis of acridine orange (AO) stained germinating Bacillus subtilis var. niger bacteria revealed a cell population initially dominated by small green spores followed by the emergence of at least three additional discernible subpopulations in response to stimulation with D-glucose. These subpopulations were small, round or oblong red cells; intermediate to large metachromatic cells; and large red rods. Large green rods were rarely observed. An increase in red emissions (i.e., putative RNA synthesis) was sometimes seen as early as 90 min after exposure to D-glucose and uptake of AO at room temperature. This may represent either metabolic recovery from quiescence or RNA synthesis associated with germination. In the absence of D-glucose, or using autoclaved bacteria in the presence of glucose, no relative increase in the red signal was observed despite hours of observation. Digital image analysis was used for relative measurement of red, green and blue signals and to correlate the size of various subpopulations with their fluorescence color emissions over time. Image analysis demonstrated a trend toward increasing size and red emission in the presence of glucose. The average red emission was found to be a good discriminator of the various subpopulations, while the average green emission was approximately equal among the subpopulations making it a poor discriminator. These data suggest that AO staining might be used for rapid computer-assisted discrimination of spores vs. vegetative cells.     

Estimating the immunity coverage required to prevent epidemics in a community of households

An estimation of the immunity coverage needed to prevent future outbreaks of an infectious disease is considered for a community of households. Data on outbreak size in a sample of households from one epidemic are used to derive maximum likelihood estimates and confidence bounds for parameters of a stochastic model for disease transmission in a community of households. These parameter estimates induce estimates and confidence bounds for the basic reproduction number and the critical immunity coverage, which are the parameters of main interest when aiming at preventing major outbreaks in the future. The case when individuals are homogeneous, apart from the size of their household, is considered in detail. The generalization to the case with variable infectivity, susceptibility and/or mixing behaviour is discussed more briefly. The methods are illustrated with an application to data on influenza in Tecumseh, Michigan.