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UnlabelledCoccidioidomycosis (or valley fever) is a fungal disease with high morbidity and mortality that affects tens of thousands of people each year. This infection is caused by two sibling species, Coccidioides immitis and C. posadasii, which are endemic to specific arid locales throughout the Western Hemisphere, particularly the desert southwest of the United States. Recent epidemiological and population genetic data suggest that the geographic range of coccidioidomycosis is expanding, as new endemic clusters have been identified in the state of Washington, well outside the established endemic range. The genetic mechanisms and epidemiological consequences of this expansion are unknown and require better understanding of the population structure and evolutionary history of these pathogens. Here we performed multiple phylogenetic inference and population genomics analyses of 68 new and 18 previously published genomes. The results provide evidence of substantial population structure in C. posadasii and demonstrate the presence of distinct geographic clades in central and southern Arizona as well as dispersed populations in Texas, Mexico, South America, and Central America. Although a smaller number of C. immitis strains were included in the analyses, some evidence of phylogeographic structure was also detected in this species, which has been historically limited to California and Baja, Mexico. Bayesian analyses indicated that C. posadasii is the more ancient of the two species and that Arizona contains the most diverse subpopulations. We propose a southern Arizona-northern Mexico origin for C. posadasii and describe a pathway for dispersal and distribution out of this region.ImportanceCoccidioidomycosis, or valley fever, is caused by the pathogenic fungi Coccidioides posadasii and C. immitis The fungal species and disease are primarily found in the American desert southwest, with spotted distribution throughout the Western Hemisphere. Initial molecular studies suggested a likely anthropogenic movement of C. posadasii from North America to South America. Here we comparatively analyze eighty-six genomes of the two Coccidioides species and establish local and species-wide population structures to not only clarify the earlier dispersal hypothesis but also provide evidence of likely ancestral populations and patterns of dispersal for the known subpopulations of C. posadasii.

The emergence of distinct populations of Cryptococcus gattii in the temperate North American Pacific Northwest (PNW) was surprising, as this species was previously thought to be confined to tropical and semitropical regions. Beyond a new habitat niche, the dominant emergent population displayed increased virulence and caused primary pulmonary disease, as opposed to the predominantly neurologic disease seen previously elsewhere. Whole-genome sequencing was performed on 118 C. gattii isolates, including the PNW subtypes and the global diversity of molecular type VGII, to better ascertain the natural source and genomic adaptations leading to the emergence of infection in the PNW. Overall, the VGII population was highly diverse, demonstrating large numbers of mutational and recombinational events; however, the three dominant subtypes from the PNW were of low diversity and were completely clonal. Although strains of VGII were found on at least five continents, all genetic subpopulations were represented or were most closely related to strains from South America. The phylogenetic data are consistent with multiple dispersal events from South America to North America and elsewhere. Numerous gene content differences were identified between the emergent clones and other VGII lineages, including genes potentially related to habitat adaptation, virulence, and pathology. Evidence was also found for possible gene introgression from Cryptococcus neoformans var. grubii that is rarely seen in global C. gattii but that was present in all PNW populations. These findings provide greater understanding of C. gattii evolution in North America and support extensive evolution in, and dispersal from, South America. Importance: Cryptococcus gattii emerged in the temperate North American Pacific Northwest (PNW) in the late 1990s. Beyond a new environmental niche, these emergent populations displayed increased virulence and resulted in a different pattern of clinical disease. In particular, severe pulmonary infections predominated in contrast to presentation with neurologic disease as seen previously elsewhere. We employed population-level whole-genome sequencing and analysis to explore the genetic relationships and gene content of the PNW C. gattii populations. We provide evidence that the PNW strains originated from South America and identified numerous genes potentially related to habitat adaptation, virulence expression, and clinical presentation. Characterization of these genetic features may lead to improved diagnostics and therapies for such fungal infections. The data indicate that there were multiple recent introductions of C. gattii into the PNW. Public health vigilance is warranted for emergence in regions where C. gattii is not thought to be endemic.

Sequence analyses and subtyping of Bacillus anthracis strains from Georgia reveal a single distinct lineage (Aust94) that is ecologically established. Phylogeographic analysis and comparisons to a global collection reveals a clade that is mostly restricted to Georgia. Within this clade, many groups are found around the country, however at least one subclade is only found in the eastern part. This pattern suggests that dispersal into and out of Georgia has been rare and despite historical dispersion within the country, for at least for one lineage, current spread is limited.

Recently, some commercial apple growers have been adopting hedging as an alternative or supplement to hand-pruning. With increasing labor costs across the United States, alternatives to hand-pruning and current training systems are being considered. One management strategy involves transitioning tall spindle trees to a narrow tree wall and simplifying labor-intensive activities of apple production, such as pruning, harvesting, and fruit thinning. The objective is to form the orchard system into a "fruiting wall" that makes fruit more visible and accessible, thus facilitating harvesting. Four management practices (tall spindle; narrow tree wall with manual pruning; narrow tree wall with dormant and summer hedging; and narrow tree wall with dormant hedging, summer hedging, and root pruning) were used to convert 12-year-old 'Brak Fuji' apple trees from the tall spindle training system to a narrow tree wall. Photosynthetically active radiation (PAR) and ultraviolet light levels within the canopies were improved by summer hedging, but they were still low for all treatments. Light within the canopy was improved most when root pruning was included. Specific leaf weight was not significantly impacted by hedging or root pruning. Detailed spur sampling showed that treatments had no effect on vegetative or reproductive growth on 2- to 3-year-old wood. Although dormant plus summer hedging alone did not affect shoot length, the combination of hedging and root pruning caused a significant reduction in terminal shoot length. Red fruit color was only improved with dormant hedging plus summer hedging plus root pruning. Compared with dormant plus summer hedging, dormant plus summer hedging plus root pruning improved fruit set and yield, but it reduced fruit size. Without root pruning, hedging had little effect on light, specific leaf weight, flower initiation, fruit set, and fruit quality. Conversion to narrow tree walls by manual pruning resulted in more poorly colored fruit and less highly colored fruit compared with maintaining the trees as tall spindles with manual pruning. [ABSTRACT FROM AUTHOR]

Tularemia, caused by the bacterium Francisella tularensis, is a potentially severe disease that often causes unspecific symptoms; because of its low infectious dose and ease of dissemination, F. tularensis is [...]

Bacillus anthracis is a gram-positive endosporeforming bacterium that causes the disease anthrax in livestock, wildlife, and humans. Because of its hardy spores, B. anthracis can survive for extended periods in [...]

Genotyping of microorganisms typically relies on comparison of genomic features (e.g., fragment size, repeats, single-nucleotide polymorphisms [SNPs]) between strains and/or against a database of feature profiles (e.g., PulseNET and mlst.net) [...]

A small number of conserved canonical single nucleotide polymorphisms (canSNP) that define major phylogenetic branches for Bacillus anthracis were used to place a Sverdlovsk patient's B. anthracis genotype into 1 [...]

Burkholderia pseudomallei soil isolates from northeast Thailand are genotyped using multiple-locus variable number tandem repeat analysis (MLVA) and multilocus sequence typing (MLST). MLVA identified 19 genotypes within three clades, while MLST revealed two genotypes.

Research has been conducted on the homogeneous bacteria Bacillus anthracis. The discovery of the ca. 750-nucleotide open reading frame containing in-frame variable-number tandem-repeat sequences is described.

A bacterial biosensor for benzene, toluene and similar compounds appears to be capable of accurately measuring environmental contaminants, making it a potentially inexpensive tool for field-ready assays. A reporter plasmid was constructed by placing the luc gene for firefyl luciferase directly under the control of the P(sub u) promoter and the xylR transcriptional activator of Pseudomonas putida mt-2. The biosensor cells were shown to detect benzene, toluene and xylene contamination in soil and water samples with high sensitivity and quantitative accuracy.

Genetic marker systems have improved dramatically in the past 10 yr. Each new system needs to be evaluated for its distribution of markers on genetic linkage maps to validate its use for genetic analysis. The resulting maps an also useful for establishing the genetic positions of genes affecting important phenotypes. We have constructed a high density map in soybean [Glycine max (L.) Merr.] using a 300 RIL (recombinant inbred line) population from BSR101 x PI437.654 by first constructing an RFLP (restriction fragment length polymorphism) 'scaffold' map based on the entire population. The RFLP anchored map was then further populated with AFLP (amplified fragment length polymorphism) markers based on only a 42 RIL subset. We report here an 840 marker map consisting of 165 RFLP, 25 RAPD (random amplified polymorphic DNA), and 650 AFLP markers spread over 29 linkage groups representing 3441 cM distance. Although clustering of AFLP markers occurred, markers were mapped to every linkage group and were well distributed relative to other marker systems. The AFLP marker system appears to be a useful approach for generating high density genetic maps in soybean., GENETIC MAPPING has been greatly enhanced in the last decade with the introduction of novel genetic marker systems. These systems have resulted in increased capabilities for detecting DNA polymorphism and [...]

Amplified fragment length polymorphism (AFLP) markers were utilized to determine the phylogenetic relationship between Bacillus anthracis and related Bacillus species. Polymerase chain reaction analysis of the Bacillus genome with AFLP markers indicated the presence of low levels of molecular variation in Bacillus anthracis due to low rates of evolution that restricted the generation of molecular diversity. Furthermore, genomic analysis indicated the close phylogenetic relationship between Bacillus anthracis and Bacillus cereus.

Abstract Background An isolate originally labeled Bacillus megaterium CDC 684 was found to contain both pXO1 and pXO2, was non-hemolytic, sensitive to gamma-phage, and produced both the protective antigen and the poly-D-glutamic acid capsule. These phenotypes prompted Ezzell et al., (J. Clin. Microbiol. 28:223) to reclassify this isolate to Bacillus anthracis in 1990. Results We demonstrate that despite these B. anthracis features, the isolate is severely attenuated in a guinea pig model. This prompted whole genome sequencing and closure. The comparative analysis of CDC 684 to other sequenced B. anthracis isolates and further analysis reveals: a) CDC 684 is a close relative of a virulent strain, Vollum A0488; b) CDC 684 defines a new B. anthracis lineage (at least 51 SNPs) that includes 15 other isolates; c) the genome of CDC 684 contains a large chromosomal inversion that spans 3.3 Mbp; d) this inversion has caused a displacement of the usual spatial orientation of the origin of replication (ori) to the termination of replication (ter) from 180° in wild-type B. anthracis to 120° in CDC 684 and e) this isolate also has altered growth kinetics in liquid media. Conclusions We propose two alternative hypotheses explaining the attenuated phenotype of this isolate. Hypothesis 1 suggests that the skewed ori/ter relationship in CDC 684 has altered its DNA replication and/or transcriptome processes resulting in altered growth kinetics and virulence capacity. Hypothesis 2 suggests that one or more of the single nucleotide polymorphisms in CDC 684 has altered the expression of a regulatory element or other genes necessary for virulence.

Abstract Background The global pattern of distribution of 1033 B. anthracis isolates has previously been defined by a set of 12 conserved canonical single nucleotide polymorphisms (canSNP). These studies reinforced the presence of three major lineages and 12 sub-lineages and sub-groups of this anthrax-causing pathogen. Isolates that form the A lineage (unlike the B and C lineages) have become widely dispersed throughout the world and form the basis for the geographical disposition of "modern" anthrax. An archival collection of 191 different B. anthracis isolates from China provides a glimpse into the possible role of Chinese trade and commerce in the spread of certain sub-lineages of this pathogen. Canonical single nucleotide polymorphism (canSNP) and multiple locus VNTR analysis (MLVA) typing has been used to examine this archival collection of isolates. Results The canSNP study indicates that there are 5 different sub-lineages/sub-groups in China out of 12 previously described world-wide canSNP genotypes. Three of these canSNP genotypes were only found in the western-most province of China, Xinjiang. These genotypes were A.Br.008/009, a sub-group that is spread across most of Europe and Asia; A.Br.Aust 94, a sub-lineage that is present in Europe and India, and A.Br.Vollum, a lineage that is also present in Europe. The remaining two canSNP genotypes are spread across the whole of China and belong to sub-group A.Br.001/002 and the A.Br.Ames sub-lineage, two closely related genotypes. MLVA typing adds resolution to the isolates in each canSNP genotype and diversity indices for the A.Br.008/009 and A.Br.001/002 sub-groups suggest that these represent older and established clades in China. Conclusion B. anthracis isolates were recovered from three canSNP sub-groups (A.Br.008/009, A.Br.Aust94, and A.Br.Vollum) in the western most portion of the large Chinese province of Xinjiang. The city of Kashi in this province appears to have served as a crossroads for not only trade but the movement of diseases such as anthrax along the ancient "silk road". Phylogenetic inference also suggests that the A.Br.Ames sub-lineage, first identified in the original Ames strain isolated from Jim Hogg County, TX, is descended from the A.Br.001/002 sub-group that has a major presence in most of China. These results suggest a genetic discontinuity between the younger Ames sub-lineage in Texas and the large Western North American sub-lineage spread across central Canada and the Dakotas.