Comparative genomic analysis reveals evidence of two novel Vibrio species closely related to V. cholerae

Background: In recent years genome sequencing has been used to characterize new bacterial species, a method of analysis available as a result of improved methodology and reduced cost. Included in a constantly expanding list of Vibrio species are several that have been reclassified as novel members of the Vibrionaceae. The description of two putative new Vibrio species, Vibrio sp. RC341 and Vibrio sp. RC586 for which we propose the names V. metecus and V. parilis, respectively, previously characterized as non-toxigenic environmental variants of V. cholerae is presented in this study. Results: Based on results of whole-genome average nucleotide identity (ANI), average amino acid identity (AAI), rpoB similarity, MLSA, and phylogenetic analysis, the new species are concluded to be phylogenetically closely related to V. cholerae and V. mimicus. Vibrio sp. RC341 and Vibrio sp. RC586 demonstrate features characteristic of V. cholerae and V. mimicus, respectively, on differential and selective media, but their genomes show a 12 to 15% divergence (88 to 85% ANI and 92 to 91% AAI) compared to the sequences of V. cholerae and V. mimicus genomes (ANI <95% and AAI <96% indicative of separate species). Vibrio sp. RC341 and Vibrio sp. RC586 share 2104 ORFs (59%) and 2058 ORFs (56%) with the published core genome of V. cholerae and 2956 (82%) and 3048 ORFs (84%) with V. mimicus MB-451, respectively. The novel species share 2926 ORFs with each other (81% Vibrio sp. RC341 and 81% Vibrio sp. RC586). Virulence-associated factors and genomic islands of V. cholerae and V. mimicus, including VSP-I and II, were found in these environmental Vibrio spp. Conclusions: Results of this analysis demonstrate these two environmental vibrios, previously characterized as variant V. cholerae strains, are new species which have evolved from ancestral lineages of the V. cholerae and V. mimicus clade. The presence of conserved integration loci for genomic islands as well as evidence of horizontal gene transfer between these two new species, V. cholerae, and V. mimicus suggests genomic islands and virulence factors are transferred between these species.

Distribution, Diversity, and Seasonality of Waterborne Salmonellae in a Rural Watershed▿

Salmonella outbreaks from contaminated water and nonanimal foods (e.g., produce) are increasingly reported. To address the environment as a potential source of pathogenic Salmonella, we investigated levels of salmonellae and the geographic and temporal variation of Salmonella serotypes from surface waters in a region of Georgia (United States) with a history of high salmonellosis case rates. Monthly water samples were collected from six stations in the Little River (Upper Suwannee Basin) for 12 months (April 2005 to April 2006). Salmonellae were enumerated using a three-step most-probable-number (MPN) assay. Salmonellae were detected in 57 of the 72 water samples collected (79.2%). Monthly Salmonella densities ranged from an MPN of 2.5 liter−1 in April 2005 to 36.3 liter−1 in August 2005; concentrations were significantly higher in the summer months compared to other seasons (P < 0.05). Concentrations were not significantly different between stations. Levels of salmonellae were correlated with average daily watershed rainfall for the 1 and 2 days preceding each sample collection (r = 0.77 and 0.68, respectively; P < 0.005). Additionally, water temperature was also positively associated with total Salmonella levels (r = 0.44; P < 0.05). In total, 13 S. enterica serotypes were identified among 197 Salmonella isolates. Eighty (40.6%) were identified as S. enterica subsp. arizonae. Muenchen and Rubislaw were the most frequently identified serotypes of the remaining 117 isolates (28 and 26 isolates, respectively). Serotype diversity peaked in the summer, with 9 serotypes observed in August compared to only one serotype (S. enterica subsp. arizonae) observed in April (2005 and 2006) (P < 0.05). Furthermore, all samples collected in August (6/6) contained multiple serotypes (two to five per sample). The results of this study suggest that Salmonella abundance and diversity in the environment vary temporally and are strongly influenced by seasonal precipitation and water temperature

Complete Closed Genome Sequences of Three Salmonella enterica subsp. enterica Serovar Dublin Strains Isolated from Cattle at Harvest

Salmonella enterica subsp. enterica serovar Dublin is a host-adapted pathogen for cattle that can cause invasive disease in humans. To facilitate genomic comparisons characterizing virulence determinants of this pathogen, we present the complete genome sequences of three S. Dublin strains isolated from bovine sources at harvest

Discovery of novel Vibrio cholerae VSP-II genomic islands using comparative genomic analysis

This report describes Vibrio seventh pandemic island II (VSP-II) and three novel variants revealed by comparative genomics of 23 Vibrio cholerae strains and their presence among a large and diverse collection of V. cholerae isolates. Three VSP-II variants were reported previously and our results demonstrate the presence of three novel VSP-II in clinical and environmental V. cholerae marked by major deletions and genetic rearrangements. A new VSP-II cluster was found in the seventh pandemic V. cholerae O1 El Tor strain CIRS101, which is dominant (95%) among the recent (2004-2007) seven pandemic V. cholerae O1 El Tor isolates from two endemic sites, but was not found in older strains from the same region. Two other variants were found in V. cholerae TMA21 and RC385, two environmental strains from coastal Brazil and the Chesapeake Bay, respectively, the latter being prevalent among environmental V. cholerae non-O1/non-O139 and Vibrio mimicus. The results of this study indicate that the VSP-II island has undergone significant rearrangement through a complex evolutionary pathway in V. cholerae. Interestingly, one of the new VSP-II revealed the presence of 'old' and 'new' V. cholerae O1 El Tor pandemic clones circulating in some of the areas where cholera is endemic.

Alternative Growth Promoters Modulate Broiler Gut Microbiome and Enhance Body Weight Gain

Antibiotic growth promoters (AGPs) are frequently used to enhance weight-gain in poultry production. However, there has been increasing concern over the impact of AGP on the emergence of antibiotic resistance in zoonotic bacterial pathogens in the microbial community of the poultry gut. In this study, we adopted mass-spectrophotometric, phylogenetic, and shotgun-metagenomic approaches to evaluate bioactive phenolic extracts (BPE) from blueberry (Vaccinium corymbosum) and blackberry (Rubus fruticosus) pomaces as AGP alternatives in broilers. We conducted two trials with 100 Cobb-500 broiler chicks (in each trial) in four equal groups that were provided water with no supplementation, supplemented with AGP (tylosin, neomycin sulfate, bacitracin, erythromycin, and oxytetracycline), or supplemented with 0.1 g Gallic acid equivalent (GAE)/L or 1.0 g GAE/L (during the last 72 h before euthanasia) of BPE for 6 weeks. When compared with the control group (water only), the chickens supplemented with AGP and 0.1 g GAE/L of BPE gained 9.5 and 5.8% more body weight, respectively. The microbiomes of both the AGP- and BPE-treated chickens had higher Firmicutes to Bacteroidetes ratios. AGP supplementation appeared to be associated with higher relative abundance of bacteriophages and unique cecal resistomes compared with BPE supplementation or control. Functional characterization of cecal microbiomes revealed significant animal-to-animal variation in the relative abundance of genes involved in energy and carbohydrate metabolism. These findings established a baseline upon which mechanisms of plant-based performance enhancers in regulation of animal growth can be investigated. In addition, the data will aid in designing alternate strategies to improve animal growth performance and consequently production

Genomic and Evolutionary Analysis of Two <i>Salmonella enterica</i> Serovar Kentucky Sequence Types Isolated from Bovine and Poultry Sources in North America

<div><p><i>Salmonella enterica</i> subsp. <i>enterica</i> serovar Kentucky is frequently isolated from healthy poultry and dairy cows and is occasionally isolated from people with clinical disease. A genomic analysis of 119 isolates collected in the United States from dairy cows, ground beef, poultry and poultry products, and human clinical cases was conducted. Results of the analysis demonstrated that the majority of poultry and bovine-associated <i>S</i>. Kentucky were sequence type (ST) 152. Several bovine-associated (n = 3) and food product isolates (n = 3) collected from the United States and the majority of human clinical isolates were ST198, a sequence type that is frequently isolated from poultry and occasionally from human clinical cases in Northern Africa, Europe and Southeast Asia. A phylogenetic analysis indicated that both STs are more closely related to other <i>Salmonella</i> serovars than they are to each other. Additionally, there was strong evidence of an evolutionary divergence between the poultry-associated and bovine-associated ST152 isolates that was due to polymorphisms in four core genome genes. The ST198 isolates recovered from dairy farms in the United States were phylogenetically distinct from those collected from human clinical cases with 66 core genome SNPs differentiating the two groups, but more isolates are needed to determine the significance of this distinction. Identification of <i>S</i>. Kentucky ST198 from dairy animals in the United States suggests that the presence of this pathogen should be monitored in food-producing animals.</p></div

SNP differences between 198.1 (non-clinical North American agricultural isolates) and 198.2 (human clinical isolates).

<p>The reference genome for this analysis is the chromosome of <i>S</i>. Kentucky CVM29188.</p

Phylogenetic relationships of <i>S</i>. Kentucky ST152 and ST198 with representatives of subclade A1 serovars as described by Timme et al.

<p>[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0161225#pone.0161225.ref016" target="_blank">16</a>] inferred using the Maximum Likelihood method with the General Time Reversible model of nucleotide substitution. Bar length represents number of substitutions per site.</p