Current methods struggle to reconstruct and visualize the genomic relationships of large numbers of bacterial genomes. GrapeTree facilitates the analyses of large numbers of allelic profiles by a ...static "GrapeTree Layout" algorithm that supports interactive visualizations of large trees within a web browser window. GrapeTree also implements a novel minimum spanning tree algorithm (MSTree V2) to reconstruct genetic relationships despite high levels of missing data. GrapeTree is a stand-alone package for investigating phylogenetic trees plus associated metadata and is also integrated into EnteroBase to facilitate cutting edge navigation of genomic relationships among bacterial pathogens.
High-throughput sequencing methods generated allele and single nucleotide polymorphism information for thousands of bacterial strains that are publicly available in online repositories and created ...the possibility of generating similar information for hundreds to thousands of strains more in a single study. Minimum spanning tree analysis of allelic data offers a scalable and reproducible methodological alternative to traditional phylogenetic inference approaches, useful in epidemiological investigations and population studies of bacterial pathogens. PHYLOViZ Online was developed to allow users to do these analyses without software installation and to enable easy accessing and sharing of data and analyses results from any Internet enabled computer. PHYLOViZ Online also offers a RESTful API for programmatic access to data and algorithms, allowing it to be seamlessly integrated into any third party web service or software. PHYLOViZ Online is freely available at https://online.phyloviz.net.
One Gram-stain-positive, non-motile, non-spore-forming, catalase-negative, and coccobacilli-shaped strain, designated c10Ua161M
, was isolated from a urine sample from a reproductive-age healthy ...woman. Comparative 16S rRNA gene sequence analysis indicated that strain c10Ua161M
belonged to the genus
. Phylogenetic analysis based on
and
gene sequences strongly supported a clade encompassing strains c10Ua161M
and eight other strains from public databases, distinct from currently recognized species of the genus
Average Nucleotide Identity (ANI) and Genome-to-Genome Distance Calculator (GGDC), showed 87.9 and 34.3 % identity to the closest relative
, respectively. The major fatty acids of strain c10Ua161M
were C
ω
(65.0%), C
(17.8%), and summed feature 8 (10.2 %; comprising C
ω7
, and/or C
ω6
). The DNA G+C content of the strains is 34.2 mol%. On the basis of data presented here, strain c10Ua161M
represents a novel species of the genus
, for which the name
sp. nov. is proposed. The type strain is c10Ua161M
(=CECT 9755
=DSM 108704
).
We characterised Lancefield group B streptococcal (GBS) isolates causing invasive disease among non-pregnant adults in Portugal between 2009 and 2015. All isolates (n = 555) were serotyped, assigned ...to clonal complexes (CCs) by multilocus sequence typing and characterised by surface protein and pilus island gene profiling. Antimicrobial susceptibility was tested by disk diffusion and resistance genotypes identified by PCR. Overall, serotype Ia was most frequent in the population (31%), followed by serotypes Ib (24%) and V (18%). Serotype Ib increased significantly throughout the study period (p < 0.001) to become the most frequent serotype after 2013. More than 40% of isolates clustered in the CC1/
3/PI-1+PI-2a genetic lineage, including most isolates of serotypes Ib (n = 110) and V (n = 65). Erythromycin and clindamycin resistance rates were 35% and 34%, respectively, both increasing from 2009 to 2015 (p < 0.010) and associated with CC1 and serotype Ib (p < 0.001). The Ib/CC1 lineage probably resulted from acquisition of the type Ib capsular operon in a single recombination event by a representative of the V/CC1 macrolide-resistant lineage. Expansion of the new serotype Ib/CC1 lineage resulted in increased macrolide resistance in GBS, causing invasive disease among adults in Portugal. The presence of this clone elsewhere may predict more widespread increase in resistance.
Abstract
Objectives
Streptococcus agalactiae group B streptococci (GBS) have been considered uniformly susceptible to penicillin. However, increasing reports from Asia and North America are ...documenting penicillin-non-susceptible GBS (PRGBS) with mutations in pbp genes. Here we report, to the best of our knowledge, the first two PRGBS isolates recovered in Europe (AC-13238-1 and AC-13238-2), isolated from the same patient.
Methods
Two different colony morphologies of GBS were noted from a surgical abscess drainage sample. Both were serotyped and antimicrobial susceptibility testing was performed by different methodologies. High-throughput sequencing was done to compare the isolates at the genomic level, to identify their capsular type and ST, to evaluate mutations in the pbp genes and to compare the isolates with the genomes of other PRGBS isolates sharing the same serotype and ST.
Results
Isolates AC-13238-1 and AC-13238-2 presented MICs above the EUCAST and CLSI breakpoints for penicillin susceptibility. Both shared the capsular type Ia operon and ST23. Genomic analysis uncovered differences between the two isolates in seven genes, including altered pbp genes. Deduced amino acid sequences revealed critical substitutions in PBP2X in both isolates. Comparison with serotype Ia clonal complex 23 PRGBS from the USA reinforced the similarity between AC-13238-1 and AC-13238-2, and their divergence from the US strains.
Conclusions
Our results support the in-host evolution of β-lactam-resistant GBS, with two PRGBS variants being isolated from one patient.
Although serogroup 6 was among the first to be recognized among Streptococcus pneumoniae, several new serotypes were identified since the introduction of pneumococcal conjugate vaccines (PCVs). A ...decrease of the 6B-2 variant among invasive pneumococcal disease (IPD), but not 6B-1, was noted post conjugate vaccine introduction, underpinned by a decrease of CC273 isolates. Serotype 6C was associated with adult IPD and increased in this age group representing two lineages (CC315 and CC395), while the same lineages expressed other serogroup 6 serotypes in children. Taken together, these findings suggest a potential cross-protection of PCVs against serotype 6C IPD among vaccinated children but not among adults. Serotype 6A became the most important serogroup 6 serotype in children but it decreased in adult IPD. No other serogroup 6 serotypes were detected, so available phenotypic or simple genotypic assays remain adequate for distinguishing serotypes within serogroup 6 isolates.
The interaction between bacteriophages, bacteria and the human host as a tripartite system has recently captured attention. The taxonomic diversity of bacteriophages, as a natural parasite of ...bacteria, still remains obscure in human body biomes, representing a so-called “viral dark matter.” Here, we isolated and characterized coliphages from blood, urine and tracheal aspirates samples collected at a tertiary care hospital in Austria. Phages were more often isolated from blood, followed by urine and tracheal aspirates. Phylogenetic analysis and genome comparisons allowed the identification of phages belonging to the
Tunavirinae
subfamily, and to the
Peduovirus
and
Tequintavirus
genera.
Tunavirinae
phages cluster together and are found in samples from 14 patients, suggesting their prevalence across a variety of human samples. When compared with other phage genomes, the highest similarity level was at 87.69% average nucleotide identity (ANI), which suggests that these are in fact a newly isolated phage species.
Tequintavirus
phages share a 95.90% with phage 3_29, challenging the ANI threshold currently accepted to differentiate phage species. The isolated phages appear to be virulent, with the exception of the
Peduovirus
members, which are integrative and seem to reside as prophages in bacterial genomes.
Polymorphic toxins (PTs) are multi-domain bacterial exotoxins belonging to distinct families that share common features in terms of domain organization. PTs are found in all major bacterial clades, ...including many toxic effectors of type V and type VI secretion systems. PTs modulate the dynamics of microbial communities by killing or inhibiting the growth of bacterial competitors lacking protective immunity proteins.
In this work, we identified a novel widespread family of PTs, named MuF toxins, which were exclusively encoded within temperate phages and their prophages. By analyzing the predicted proteomes of 1845 bacteriophages and 2464 bacterial genomes, we found that MuF-containing proteins were frequently part of the DNA packaging module of tailed phages. Interestingly, MuF toxins were abundant in the human gut microbiome.
Our results uncovered the presence of the MuF toxin family in the temperate phages of Firmicutes. The MuF toxin family is likely to play an important role in the ecology of the human microbiota where pathogens and commensal species belonging to the Firmicutes are abundant. We propose that MuF toxins could be delivered by phages into host bacteria and either influence the lysogeny decision or serve as bacterial weapons by inhibiting the growth of competing bacteria.
The rise of antibiotic resistance in pathogenic bacteria is a significant problem for the treatment of infectious diseases. Resistance is usually selected by the antibiotic itself; however, biocides ...might also co-select for resistance to antibiotics. Although resistance to biocides is poorly defined, different in vitro studies have shown that mutants presenting low susceptibility to biocides also have reduced susceptibility to antibiotics. However, studies with natural bacterial isolates are more limited and there are no clear conclusions as to whether the use of biocides results in the development of multidrug resistant bacteria.
The main goal is to perform an unbiased blind-based evaluation of the relationship between antibiotic and biocide reduced susceptibility in natural isolates of Staphylococcus aureus. One of the largest data sets ever studied comprising 1632 human clinical isolates of S. aureus originated worldwide was analysed. The phenotypic characterization of 13 antibiotics and 4 biocides was performed for all the strains. Complex links between reduced susceptibility to biocides and antibiotics are difficult to elucidate using the standard statistical approaches in phenotypic data. Therefore, machine learning techniques were applied to explore the data.
In this pioneer study, we demonstrated that reduced susceptibility to two common biocides, chlorhexidine and benzalkonium chloride, which belong to different structural families, is associated to multidrug resistance. We have consistently found that a minimum inhibitory concentration greater than 2 mg/L for both biocides is related to antibiotic non-susceptibility in S. aureus.
Two important results emerged from our work, one methodological and one other with relevance in the field of antibiotic resistance. We could not conclude on whether the use of antibiotics selects for biocide resistance or vice versa. However, the observation of association between multiple resistance and two biocides commonly used may be of concern for the treatment of infectious diseases in the future.
A epidemiologia das infecções por Staphylococcus aureus resistente a meticilina (MRSA) é dinâmica, havendo aumento na prevalência de MRSA ocasionando infecções adquiridas na comunidade (CA) ou ...relacionadas à assistência à saúde (IRAS). O objetivo deste estudo foi avaliar a epidemiologia de infecções de corrente sanguínea ocasionadas por MRSA e avaliar os fatores de risco associados a mortalidade.
Estudo de coorte, realizado em um complexo hospitalar da cidade de Porto Alegre (RS), entre abril de 2014 a junho de 2018. Os isolados foram avaliados por high-throughput sequencing (tecnologia Illumina) e as informações para tipagem foram obtidas. Teste de suscetibilidade aos antimicrobianos foram realizados por disco-difusão, conforme protocolos do EUCAST e a concentração inibitória mínima para vancomicina (CIM) foi determinada por fitas de gradiente. Dados clínicos e epidemiológicos dos pacientes foram obtidos através do prontuário eletrônico.
Foram incluídos 91 pacientes. A média de idade foi 52,4 anos (DP +-23,1) e as comorbidades mais frequentes foram hipertensão (39,6%), diabete mellitus (37,4%) e doença cardiovascular (18,7%). A mortalidade intra-hospitalar foi de 33,0%. Não houve diferenças significativas em características epidemiológicas considerando a origem das infecções (AC x IRAS). A CIM para vancomicina variou de 0,5mg/L a 2,0mg/L com 59,3% dos isolados apresentado CIM ≥ 1,5mg/L. Um total de 29 (31,9%) isolados foram classificados como multidrogarresistente, sendo mais prevalentes entre às IRAS (p = 0,001). Considerando as sequencias tipo (ST) e complexos clonais (CC), ambos ST30 e CC30 estavam associados a infecções AC (p = 0,022 e p = 0.006). O CC5 estava associado a IRAS (p = 0,008), com todas as suas STs (ST105, ST1176, ST1635 e ST5521) estando relacionadas a IRAS. Considerando os fatores de risco para mortalidade: idade (p < 0,001), doença cardiovascular (p = 0,005) e elevado Charlson Comorbidity Index (p = 0,001) estavam associados a mortalidade na análise univariada, mas somente idade foi um de risco independente para mortalidade (IC 1.004-1.043; p = 0,017). Não foi evidenciada associação de CC, nem de características fenotípicas ou genotípicos com a mortalidade.
Foi observada elevada mortalidade intra-hospitalar e estava associada com a maiores idades. Além de não haver associação entre os CC e mortalidade, foi evidenciado que o CC30 (ST30-mecIVc-PVL+) foi o clone dominante entre as infeções AC.