Maximum Clade Credibility Tree Research Articles

Systematics of Urostephanus: resurrection of a Mesoamerican taxon of Gonolobinae (Apocynaceae, Asclepiadoideae), with eight new combinations

Background and Aims: Recent studies have focused on resolving the systematics of the tribe Gonolobinae of the family Apocynaceae, as well as the controversial genus Matelea s.l. However, many species within Matelea s.l. have not been evaluated phylogenetically. This is the case of M. gonoloboides, previously recognized as the type species of Urostephanus, and other taxa with similar floral morphology. The aim of this study is to test the monophyly of Urostephanus, including some species with morphology similar to M. gonoloboides. Furthermore, we evaluate the taxonomic position of the Urostephanus clade within the subtribe Gonolobinae and compare morphology among related clades. Methods: To assess phylogenetic relationships within Gonolobinae, we downloaded sequences from four loci for 94 species of the subtribe from GenBank. We extracted DNA and sequenced the trnL-F intergenic spacer and rps16 intron from four species of Dictyanthus and six species of Matelea, which are considered to belong to Urostephanus. We reconstructed the phylogeny with Bayesian inference using the maximum clade credibility tree. Based on the phylogenetic pattern, we performed morphological comparisons between the clades Dictyanthus, Polystemma, and Urostephanus. Key results: Phylogenetic analysis recovered Urostephanus as monophyletic and sister to Dictyanthus. Based on this phylogenetic pattern and floral morphology, we resurrect the genus Urostephanus. We propose eight new combinations for the species included in the phylogeny, as well as for those that share floral morphological similarity. We designate two lectotypes. Finally, we discuss aspects of morphology between Urostephanus and closely related groups. Conclusions: The current circumscription of Matelea s.l. includes species with high morphological diversity. With the phylogenetic and morphological evidence among related taxa, we support the resurrection of Urostephanus, more than 120 years after its description. These results contribute to the resolution of the systematics of Gonolobinae in Mesoamerica.

Spatial–temporal transmission dynamics of HIV-1 CRF01_AE in Indonesia

Human immunodeficiency virus type 1 (HIV-1) remains a serious health threat in Indonesia. In particular, the CRF01_AE viruses were the predominant HIV-1 strains in various cities in Indonesia. However, information on the dynamic transmission characteristics and spatial–temporal transmission of HIV-1 CRF01_AE in Indonesia is limited. Therefore, the present study examined the spatial–temporal transmission networks and evolutionary characteristics of HIV-1 CRF01_AE in Indonesia. To clarify the epidemiological connection between CRF01_AE outbreaks in Indonesia and the rest of the world, we performed phylogenetic studies on nearly full genomes of CRF01_AE viruses isolated in Indonesia. Our results showed that five epidemic clades, namely, IDN clades 1–5, of CRF01_AE were found in Indonesia. To determine the potential source and mode of transmission of CRF01_AE, we performed Bayesian analysis and built maximum clade credibility trees for each clade. Our study revealed that CRF01_AE viruses were commonly introduced into Indonesia from Southeast Asia, particularly Thailand. The CRF01_AE viruses might have spread through major pandemics in Asian countries, such as China, Vietnam, and Laos, rather than being introduced directly from Africa in the early 1980s. This study has major implications for public health practice and policy development in Indonesia. The contributions of this study include understanding the dynamics of HIV-1 transmission that is important for the implementation of HIV disease control and prevention strategies in Indonesia.

Variational Bayesian phylogenies through matrix representation of tree space.

In this article, we study the distance matrix as a representation of a phylogeny by way of hierarchical clustering. By defining a multivariate normal distribution on (a subset of) the entries in a matrix, this allows us to represent a distribution over rooted time trees. Here, we demonstrate tree distributions can be represented accurately this way for a number of published tree distributions. Though such a representation does not map to unique trees, restriction to a subspace, in particular one we call a "cube", makes the representation bijective at the cost of not being able to represent all possible trees. We introduce an algorithm "cubeVB" specifically for cubes and show through well calibrated simulation study that it is possible to recover parameters of interest like tree height and length. Although a cube cannot represent all of tree space, it is a great improvement over a single summary tree, and it opens up exciting new opportunities for scaling up Bayesian phylogenetic inference. We also demonstrate how to use a matrix representation of a tree distribution to get better summary trees than commonly used maximum clade credibility trees. An open source implementation of the cubeVB algorithm is available from https://github.com/rbouckaert/cubevb as the cubevb package for BEAST 2.

The influence of climate oscillations and geological events on population differentiation of Camponotus japonicus in the Chinese mainland.

Camponotus japonicus (Hymenoptera: Formicidae) is an omnivorous social insect which builds sizable colonies in sparse woodlands or cropland and spreads across multiple climatic zones in the Chinese mainland. This study aims to reveal the role of climate changes and geological events in driving the genetic structure of social insect populations by investigating the phylogenetics and historical demography of C. japonicus in the Chinese mainland. Phylogenetic analyses were conducted based on the mitochondria DNA dataset using MrBayes and IQ-TREE. We constructed a haplotype network, calculated analyses of molecular variance, estimated the divergence time, and reconstructed the maximum clade credibility tree. Mismatch distribution and Bayesian skyline plots were used to infer historical population fluctuations. Additionally, ecological niche modeling was employed to predict the potential distribution of the species during the present, mid-holocene, and last glacial maximum periods in the Chinese mainland. The phylogenetic tree and median-joining network analyses support the presence of four distinct lineages in C. japonicus. These lineages exhibit significant genetic differentiation and limited gene flow. The divergence among the four lineages began in the early Pleistocene, approximately 1.41 million years ago (Ma). Subsequently, the central lineage diverged from both the northern and southern lineages around 1.16 Ma, while the northern and southern lineages diverged from each other at approximately 1.07 Ma. Population expansion was observed in the southern, central, and northern lineages prior to the last glacial maximum, while the Yunnan-Sichuan lineage experienced a slight increase in population size in more recent times. The predicted distribution of the species corresponds well with the actual distribution. Furthermore, the current suitable habitat areas in northern Xinjiang, southern Tibet, and the southeast coastal regions have significantly decreased compared to the last glacial maximum and the mid-holocene periods. Our results suggest that climate oscillations and geological events play an important role in driving genetic patterns and differentiation of C. japonicus. Mountain barriers isolate populations from each other, hinder the flow of genes, and effectively prevent the spread of this species. But at the same time, it also formed refugia at low altitudes areas such as Qinling-Bashan Mountains and Yanshan-Taihang Mountains and provide suitable habitats during glaciation. This study provides a good model for understanding how complex climate changes and geological events affect population genetic differentiation of social insects in the Chinese mainland.

EvoLaps 2: Advanced phylogeographic visualization.

EvoLaps is a user-friendly web application designed to visualize the spatial and temporal spread of pathogens. It takes an annotated tree as entry, such as a maximum clade credibility tree obtained through continuous phylogeographic inference. By following a 'Top-Down' reading of a tree recursively, transitions (latitude/longitude changes from a node to its children) are represented on a cartographic background using graphical paths. The complete set of paths forms the phylogeographic scenario. EvoLaps offers several features to analyze complex scenarios: (1) enhanced path display using multiple graphical variables with time-dependent gradients, (2) cross-highlighting and selection capabilities between the phylogeographic scenario and the phylogenetic tree, (3) production of specific spatio-temporal scales and synthetic views through dynamic and iterative clustering of localities into spatial clusters, (4) animation of the phylogeographic scenario using tree brushing, which can be done manually or automatically, gradually over time or at specific time intervals, and for the entire tree or a specific clade, and (5) an evolving library of additional tools. EvoLaps is freely available for use at evolaps.org.

Epidemiology and Evolutionary Dynamics of High Pathogenicity Avian Influenza (HPAI) H5N1 in Bangladesh

Understanding disease clustering and transmission patterns improves the prevention and control of disease. Herein, we described the epizootic characteristics and spatiotemporal dynamics of High Pathogenicity Avian Influenza (HPAI) H5N1 outbreaks as well as clade diversity and phylodynamics of H5N1 over time and across host species in Bangladesh. We used Moran’s I, Geary’s C, Getis-Ord Gi∗, and a space-time permutation model to analyze the spatiotemporal patterns of H5N1 outbreaks. We used Bayesian phylogenetic analysis to generate a time-scaled maximum clade credibility (MCC) tree. Our study revealed nine HPAI H5N1 epizootic waves between 2007 and 2020 that invariably peaked in the wintertime. After vaccination of poultry against H5N1 was introduced in 2012, the incidence of HPAI H5N1 outbreaks and poultry mortality decreased significantly over time. Nonetheless, our research revealed that the virus continued circulating unabatedly in Bangladesh. The various spatiotemporal analyses were identified up to nine space-time clusters across Bangladesh, with the most significant clustering and hotspots of H5N1 outbreaks in and around the district of Dhaka. Since 2007, four H5N1 clades have been detected in Bangladesh, with only clade 2.3.2.1a continuing to circulate since 2011, which was followed up by the reassorted clade 2.3.2.1a in 2012. The HA gene of the H5N1 clade 2.3.2.1a has been reassorted into at least nine subgroups (R1–R9). After 2016, the first eight groups disappeared, with only the R9 group remaining. Spatiotemporal patterns and phylodynamics of H5N1 outbreaks are crucial for developing targeted and appropriate HPAI control and prevention measures. We recommended intensive monitoring of biosecurity measures and disease records in high-priority areas, along with assessing vaccine efficacy to better control HPAI outbreaks in Bangladesh.

Epidemiology and phylodynamics of multiple clades of H5N1 circulating in domestic duck farms in different production systems in Bangladesh.

Waterfowl are considered to be natural reservoirs of the avian influenza virus (AIV). However, the dynamics of transmission and evolutionary patterns of AIV and its subtypes within duck farms in Bangladesh remain poorly documented. Hence, a cross-sectional study was conducted in nine districts of Bangladesh between 2019 and 2021, to determine the prevalence of AIV and its subtypes H5 and H9, as well as to identify risk factors and the phylodynamics of H5N1 clades circulating in domestic duck farms. The oropharyngeal and cloacal swab samples were tested for the AIV Matrix gene (M-gene) followed by H5, H7, and H9 subtypes using rRT-PCR. The exploratory analysis was performed to estimate AIV and its subtype prevalence in different production systems, and multivariable logistic regression model was used to identify the risk factors that influence AIV infection in ducks. Bayesian phylogenetic analysis was conducted to generate a maximum clade credibility (MCC) tree and the maximum likelihood method to determine the phylogenetic relationships of the H5N1 viruses isolated from ducks. AIV was detected in 40% (95% CI: 33.0-48.1) of the duck farms. The prevalence of AIV was highest in nomadic ducks (39.8%; 95% CI: 32.9-47.1), followed by commercial ducks (24.6%; 95% CI: 14.5-37.3) and backyard ducks (14.4%; 95% CI: 10.5-19.2). The H5 prevalence was also highest in nomadic ducks (19.4%; 95% CI: 14.0-25.7). The multivariable logistic regression model revealed that ducks from nomadic farms (AOR: 2.4; 95% CI: 1.45-3.93), juvenile (AOR: 2.2; 95% CI: 1.37-3.61), and sick ducks (AOR: 11.59; 95% CI: 4.82-32.44) had a higher risk of AIV. Similarly, the likelihood of H5 detection was higher in sick ducks (AOR: 40.8; 95% CI: 16.3-115.3). Bayesian phylogenetic analysis revealed that H5N1 viruses in ducks belong to two distinct clades, 2.3.2.1a, and 2.3.4.4b. The clade 2.3.2.1a (reassorted) has been evolving silently since 2015 and forming at least nine subgroups based on >90% posterior probability. Notably, clade 2.3.4.4b was introduced in ducks in Bangladesh by the end of the year 2020, which was genetically similar to viruses detected in wild birds in Japan, China, and Africa, indicating migration-associated transmission of an emerging panzootic clade. We recommend continuing AIV surveillance in the duck production system and preventing the intermingling of domestic ducks with migratory waterfowl in wetlands.

Global molecular evolution and phylogeographic analysis of barley yellow dwarf virus based on the cp and mp genes

Barley yellow dwarf virus (BYDV) has caused considerable losses in the global production of grain crops such as wheat, barley and maize. We investigated the phylodynamics of the virus by analysing 379 and 485 nucleotide sequences of the genes encoding the coat protein and movement protein, respectively. The maximum clade credibility tree indicated that BYDV-GAV and BYDV-MAV, BYDV-PAV and BYDV-PAS share the same evolutionary lineage, respectively. The diversification of BYDV arises from its adaptability to vector insects and geography. Bayesian phylogenetic analyses showed that the mean substitution rates of the coat and movement proteins of BYDV ranged from 8.327 × 10− 4 (4.700 × 10− 4–1.228 × 10− 3) and 8.671 × 10− 4 (6.143 × 10− 4–1.130 × 10− 3) substitutions/site/year, respectively. The time since the most recent common BYDV ancestor was 1434 (1040–1766) CE (Common Era). The Bayesian skyline plot (BSP) showed that the BYDV population experienced dramatic expansions approximately 8 years into the 21st century, followed by a dramatic decline in less than 15 years. Our phylogeographic analysis showed that the BYDV population originating in the United States was subsequently introduced to Europe, South America, Australia and Asia. The migration pathways of BYDV suggest that the global spread of BYDV is associated with human activities.

Global Spread Dynamics and Intrinsic Evolutionary Dynamics of Apple Chlorotic Leaf Spot Virus Whole-Genome Sequences over the Last 30 Years

Apple chlorotic leaf spot virus (ACLSV; genus Trichovirus) is an economically important virus. Approximately 101 ACLSV whole-genome sequences were obtained from NCBI and used to explore the evolutionary dynamics of ACLSV. The time to the most recent common ancestor of ACLSV based on BEAST analysis appeared on an apple host from Canada in 1918 and then spread around the world in three ways concurrent with host spread. The maximum clade credibility tree of ACLSV shows that the mean evolution rate was 4.92 × 10-4 substitutions per site per year (subs/site/year), and we found that, during host evolution, the rate of evolution was mostly 2.31 × 10-4 to 2.72 × 10-4 subs/site/year. The rate of geographic evolution was 5.51 × 10-4 to 6.17 × 10-4 subs/site/year. To further explore the intrinsic changes in ACLSV during the process of geographic and host spread, we explored the secondary structural changes of the ACLSV coat protein (CP), which were mainly concentrated in four regions-20 to 40, 70 to 90, 120 to 140, and 180 to 193-which are related to the presence or absence and change in length of the β-turn, β strand, coil, and α helix, respectively. We then explored the codon usage preference within the CP across the migration pathways of ACLSV. These comprehensive analyses not only reveal the changes in ACLSV in the last 30 years but also further elucidate the intrinsic evolutionary dynamics of ACLSV. This is also the first report on the intrinsic evolutionary dynamics of ACLSV.

Out of Madagascar, repeatedly: The phylogenetics and biogeography of Dombeyoideae (Malvaceae s.l.)

Dispersals have been shown to be critical to the evolution of the long isolated but megadiverse flora of Madagascar and the surrounding islands of the western Indian Ocean, but we are just beginning to understand the directionality of these dispersals. With more than half of its species occurring in the western Indian Ocean region (WIOR), the paleotropical subfamily Dombeyoideae provides a particularly useful case study through which to better understand the biogeography of the WIOR, and yet its biogeography is poorly understood. Here we sampled six molecular markers from all 20 genera in the Dombeyoideae to reconstruct the most complete phylogeny to date for the subfamily. From this, divergence times, calibrated with three fossils (two dombeyoid, one malvoid), and ancestral range estimations were hypothesized. Biogeographic stochastic mapping (BSM) analyses on the maximum clade credibility tree were completed and compared to BSM analyses on 1,000 trees randomly sampled from the posterior distribution of trees resulting from the dating analysis. We found the Dombeyoideae crown node diverged ca. 53 million years ago out of a broad ancestral range involving all three major areas of its distribution: Madagascar, Africa, and Asia. The majority of diversification and dispersals in the subfamily occurred within the last ca. 10 million years, mostly from the Pliocene onwards. There were roughly five dispersals from Madagascar to Africa (and only one in reverse), at least six from Madagascar to surrounding islands of the WIOR (Mascarenes and Comoros), and one dispersal from Madagascar to Asia (and ca. 1 in reverse). Other long-distance dispersals included one from Africa to St. Helena and one from Africa to Australasia, both from within the most widespread clade, the Cheirolaena & allies clade, and one dispersal from Asia to Africa. Critically, the Dombeyoideae provide strong evidence for considering the island of Madagascar as a source for the colonization of continents, as well as the surrounding islands of the WIOR. Furthermore, narrow sympatry was a key process in the evolution of the subfamily, particularly in Madagascar and the Mascarenes.

Genetic evolution of hemagglutinin and neuraminidase genes of H5N1 highly pathogenic avian influenza viruses in Thailand.

Ongoing outbreaks of H5N1 highly pathogenic avian influenza (HPAI) viruses and the emergence of the genetic-related hemagglutinin (HA) gene of reassortant H5Nx viruses currently circulating in wild birds and poultries pose a great global public health concern. In this study, we comprehensively analyzed the genetic evolution of Thai H5N1 HA and neuraminidase (NA) genes between 2003 and 2010. The H5N1 Thailand virus clade 2.3.4 was also genetically compared to the currently circulating clade 2.3.4.4 of H5Nx viruses. Full-length nucleotide sequences of 178 HA and 143 NA genes of H5N1 viruses circulating between 2003 and 2010 were phylogenetically analyzed using maximum likelihood (ML) phylogenetic construction. Bayesian phylogenetic trees were reconstructed using BEAST analysis with a Bayesian Markov chain Monte Carlo (MCMC) approach. The maximum clade credibility (MCC) tree was determined, and the time of the most recent common ancestor (tMRCA) was estimated. The H5N1 HA nucleotide sequences of clade 2.3.4 Thailand viruses were phylogenetically analyzed using ML phylogenetic tree construction and analyzed for nucleotide similarities with various subtypes of reassortant H5Nx HA clade 2.3.4.4. ML phylogenetic analysis revealed two distinct HA clades, clade 1 and clade 2.3.4, and two distinct NA groups within the corresponding H5 clade 1 viruses. Bayesian phylogenetic reconstruction for molecular clock suggested that the Thai H5N1 HA and NA emerged in 2001.87 (95% HPD: 2001.34-2002.49) and 2002.38 (95% HPD: 2001.99-2002.82), respectively, suggesting that the virus existed before it was first reported in 2004. The Thai H5N1 HA clade 2.3.4 was grouped into corresponding clades 2.3.4, 2.3.4.1, 2.3.4.2, and 2.3.4.3, and shared nucleotide similarities to reassortant H5Nx clade 2.3.4.4 ranged from 92.4-96.8%. Phylogenetic analysis revealed monophyletic H5Nx clade 2.3.4.4 evolved from H5N1 clade 2.3.4. H5N1 viruses existed, and were presumably introduced and circulated in avian species in Thailand, before they were officially reported in 2004. HA and NA genes continuously evolved during circulation between 2004 and 2010. This study provides a better understanding of genetic evolution with respect to molecular epidemiology. Monitoring and surveillance of emerging variants/reassortants should be continued.

Unveiling new perspective of phylogeography, genetic diversity, and population dynamics of Southeast Asian and Pacific chickens

The complex geographic and temporal origins of chicken domestication have attracted wide interest in molecular phylogeny and phylogeographic studies as they continue to be debated up to this day. In particular, the population dynamics and lineage-specific divergence time estimates of chickens in Southeast Asia (SEA) and the Pacific region are not well studied. Here, we analyzed 519 complete mitochondrial DNA control region sequences and identified 133 haplotypes with 70 variable sites. We documented 82.7% geographically unique haplotypes distributed across major haplogroups except for haplogroup C, suggesting high polymorphism among studied individuals. Mainland SEA (MSEA) chickens have higher overall genetic diversity than island SEA (ISEA) chickens. Phylogenetic trees and median-joining network revealed evidence of a new divergent matrilineage (i.e., haplogroup V) as a sister-clade of haplogroup C. The maximum clade credibility tree estimated the earlier coalescence age of ancestral D-lineage (i.e., sub-haplogroup D2) of continental chickens (3.7 kya; 95% HPD 1985–4835 years) while island populations diverged later at 2.1 kya (95% HPD 1467–2815 years). This evidence of earlier coalescence age of haplogroup D ancestral matriline exemplified dispersal patterns to the ISEA, and thereafter the island clade diversified as a distinct group.

Genomic and evolutionary characteristics of G3P[8] group a rotavirus strains in China, 2016 to 2018.

Rotavirus A (RVA) G3P[8] is sporadically detected in China, although G9P[8] predominates. To evaluate their genetic composition at the whole-genome level, 24 G3P[8] RVA strains isolated from children under five years were sequenced and characterized. The 24 strains were genotyped as G3-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1, indicating the Wa-like genotype constellation. A maximum clade credibility (MCC) tree for VP7 indicated that G3 had an estimated mean evolutionary rate of 7.279×10-4 substitutions/site/year; thus, 3-5years would pass from the generation of an ancestor virus to the epidemic spread of that virus throughout China. Considering the ongoing prevalence as well as rapid evolution, it is important to monitor G3P[8] RVA epidemics; continuous nationwide surveillance is essential.

Early cephalopod evolution clarified through Bayesian phylogenetic inference

BackgroundDespite the excellent fossil record of cephalopods, their early evolution is poorly understood. Different, partly incompatible phylogenetic hypotheses have been proposed in the past, which reflected individual author’s opinions on the importance of certain characters but were not based on thorough cladistic analyses. At the same time, methods of phylogenetic inference have undergone substantial improvements. For fossil datasets, which typically only include morphological data, Bayesian inference and in particular the introduction of the fossilized birth-death model have opened new possibilities. Nevertheless, many tree topologies recovered from these new methods reflect large uncertainties, which have led to discussions on how to best summarize the information contained in the posterior set of trees.ResultsWe present a large, newly compiled morphological character matrix of Cambrian and Ordovician cephalopods to conduct a comprehensive phylogenetic analysis and resolve existing controversies. Our results recover three major monophyletic groups, which correspond to the previously recognized Endoceratoidea, Multiceratoidea, and Orthoceratoidea, though comprising slightly different taxa. In addition, many Cambrian and Early Ordovician representatives of the Ellesmerocerida and Plectronocerida were recovered near the root. The Ellesmerocerida is para- and polyphyletic, with some of its members recovered among the Multiceratoidea and early Endoceratoidea. These relationships are robust against modifications of the dataset. While our trees initially seem to reflect large uncertainties, these are mainly a consequence of the way clade support is measured. We show that clade posterior probabilities and tree similarity metrics often underestimate congruence between trees, especially if wildcard taxa are involved.ConclusionsOur results provide important insights into the earliest evolution of cephalopods and clarify evolutionary pathways. We provide a classification scheme that is based on a robust phylogenetic analysis. Moreover, we provide some general insights on the application of Bayesian phylogenetic inference on morphological datasets. We support earlier findings that quartet similarity metrics should be preferred over the Robinson-Foulds distance when higher-level phylogenetic relationships are of interest and propose that using a posteriori pruned maximum clade credibility trees help in assessing support for phylogenetic relationships among a set of relevant taxa, because they provide clade support values that better reflect the phylogenetic signal.

A Bayesian evolutionary model towards understanding wildlife contribution to F4-family Mycobacterium bovis transmission in the South-West of France

In two “départements” in the South-West of France, bovine tuberculosis (bTB) outbreaks due to Mycobacterium bovis spoligotype SB0821 have been identified in cattle since 2002 and in wildlife since 2013. Using whole genome sequencing, the aim of our study was to clarify badger contribution to bTB transmission in this area. We used a Bayesian evolutionary model, to infer phylogenetic trees and migration rates between two pathogen populations defined by their host-species. In order to account for sampling bias, sub-population structure was inferred using the marginal approximation of the structured coalescent (Mascot) implemented in BEAST2. We included 167 SB0821 strains (21 isolated from badgers and 146 from cattle) and identified 171 single nucleotide polymorphisms. We selected a HKY model and a strict molecular clock. We estimated a badger-to-cattle transition rate (median: 2.2 transitions/lineage/year) 52 times superior to the cattle-to-badger rate (median: 0.042 transitions/lineage/year). Using the maximum clade credibility tree, we identified that over 75% of the lineages from 1989 to 2000 were present in badgers. In addition, we calculated a median of 64 transition events from badger-to-cattle (IQR: 10–91) and a median of zero transition event from cattle-to-badger (IQR: 0–3). Our model enabled us to infer inter-species transitions but not intra-population transmission as in previous epidemiological studies, where relevant units were farms and badger social groups. Thus, while we could not confirm badgers as possible intermediaries in farm-to-farm transmission, badger-to-cattle transition rate was high and we confirmed long-term presence of M.bovis in the badger population in the South-West of France.

Molecular Epidemiology of SARS-CoV-2 by Sequencing.

Sequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) contains preliminary information on the phylodynamics and phylogeography of this new virus. A maximum clade credibility tree (MCCT) was constructed using available whole genome sequences of SARS-CoV-2 and highly similar whole genome sequences from bat SARS-like coronavirus, which are available in GenBank. In this chapter, we describe the molecular epidemiology of SARS-CoV-2 by sequencing the viral genomes from confirmed COVID-19 patients, utilizing methods such as target fragment amplification, sequencing, alignment, and maximum similarity mapping.

666. Molecular Epidemiology of Methicillin-Resistant Staphylococcus aureus in Chile between 1999-2018

BackgroundThe global spread of methicillin-resistant Staphylococcus aureus (MRSA) is associated with distinct genetic lineages that predominate in specific geographical regions. Available evidence suggests the Chilean-Cordobes clone (ChC), an ST5-SCCmecI lineage, has largely predominated in Chilean hospitals since its first description in the late 1990’s. Although the circulation of other MRSA lineages, including community-associated clones, has been well documented, the dynamics of clonal replacement over time has not been explored. Therefore, we aimed to study the molecular epidemiology and dynamics of clonal replacement using a large collection of clinical MRSA strains recovered from Chile during the last two decades.MethodsWe used whole-genome sequencing (WGS) and core-based phylogenomic analysis to identify genetic lineages and explore their relationship in 798 MRSA isolates obtained between 1999-2018 from two tertiary-care Chilean hospitals. ResultsOverall, the most frequently identified clones were the ST5-SCCmecI ChC (n=476, 60%), followed by ST105-SCCmecII (n=119, 15%), ST72-SCCmecIV (n=74, 9%), and ST8-SCCmecII (n=26, 3%). Phylogenomic reconstruction demonstrated 7 major clades: Clade I (CC30); Clade II (CC22); Clade III (CC97); Clade IV (CC8); Clade V (ST72); Clade VI (CC5/ST225 and ST105) and Clade VII (CC5/ST5-SCCmecI) (Fig. 1). The ChC clone remained the most frequent MRSA lineage throughout the study period (Fig. 2). However, its relative abundance decreased from >90% of isolates in 1999 to ca. 40% in 2018. This decrease began around 2005 and was associated with a progressive expansion of the ST105-SCCmecII and ST72-SCCmecIV lineages (Fig. 2). A Bayesian molecular clock analysis established the most recent common ancestor in 1964 (95% HPD interval=1961.975-1966.218) and corroborated a CC5 expansion event starting in Chile in 1999 (Fig. 3). Interestingly, our analyses revealed two branches within the ST5-SCCmecI lineage: one predominating in 1999-2006, and a more recent branch (related to the ST105-SCCmecII clone) that emerged around 2008. Figure 1. Core genome phylogenomic reconstruction of the 798 MRSA isolates. The seven major clades are represented by colored sections. The Clade I (purple section) was composed of isolates belonging to the CC30. Clade II (cyan section) includes four isolates of CC22. Clade III (red section) is composed of isolates of CC97. Clade IV (blue section) grouped isolates of different ST239 and ST8, belonging to the CC8. Clade V (orange section) includes isolates of ST72. Clade VI (yellow section) includes isolates of ST225 and ST105, both belonging to CC5. Clade VII (green section) is mostly composed of isolates of ST5-SCCmecI. The inner ring shows the ST of the isolates; the external ring shows the staphylococcal chromosomal cassette mec (SCCmec) type.Figure 2. Relative frequency of MRSA clones from 1999 to 2018. The genomes were grouped according to their isolation dates. Most frequent MRSA clones are represented by colored sections.Figure 3. Maximum clade credibility tree from the molecular clock analysis of the 798 MRSA genomes. A Bayesian molecular clock analysis was performed with BEAST using the isolation date of each genome as a calibrator. The colored strip showed the most frequent clones. The red dot shows a major event of divergence in 2008.ConclusionThe ChC clone remains the most prevalent MRSA in Chile. However, our data is consistent with the evolution of this clone and a progressive replacement of with ST105 and ST72 genetic lineages. Disclosures Lorena Diaz, PhD , Nothing to disclose Cesar A. Arias, M.D., MSc, Ph.D., FIDSA, Entasis Therapeutics (Grant/Research Support)MeMed Diagnostics (Grant/Research Support)Merk (Grant/Research Support)

728. Genomic Characterization of Burkholderia pseudomallei Isolates in Colombia

BackgroundMelioidosis is a serious infection caused by Burkholderia pseudomallei (Bps), an opportunistic organism, highly adaptable and with a wide array of intrinsic virulence factors and antimicrobial resistance determinants. Bps is underdiagnosed due to its slow growth on routine laboratory media and the lack of robust diagnostic infrastructure in rural areas of low/middle income countries. Recent data indicates that Bps infections are increasing in Colombia (COL). However, the understanding of the genomic epidemiology and population structure of the emerging Bps isolates in COL is unknown. Here we characterize the genomic features of Bps isolates from infected patients in COL.MethodsWe identified 13 Bps clinical isolates recovered in 5 Colombian cities between 2018 and 2020. We performed WGS and phylogenomic analyses using Bayesian methods. For comparisons, we included 82 publicly available genomes from Bps recovered worldwide (including 10 additional isolates from COL). Additionally, we characterized the resistome, virulome and MLST of all isolates. Results12 out of the 13 isolates were confirmed as Bps and 1 belonged to the B. cepacia complex. The Bps population structure was divided in two main clades: clade 1 with isolates from Asia and Australia, and clade 2 with isolates from Africa, America, and the Caribbean (Figure 1). We found two groups of Colombian isolates, the first was related to ST518 and the second, highly diverse including 11 different STs (1742, 1748, 92, among others). Genomic characterization showed the presence of β-lactamases PenA (n=11) and OXA-57 (n=1). We also identified a T584A substitution in PBP3 (n=11). All genomes contained virulence determinants of motility (BimA), invasion (Flagella), signaling (CdpA) and adherence (Type IV pili). Type III and VI secretion systems, were also found in all isolates resembling Bps from other parts of the world. Figure 1. Maximum clade credibility tree of 82 genomes of Bps. The inner ring shows the ST for each genome, while the outer ring shows the geographical region associated with them. Groups highlighted in red show the location of the Colombian genomes and those related to them.ConclusionBps is an emerging pathogen in COL and its population structure seems highly diverse, predominantly of the American lineage and absence of Australasians strains. A high prevalence ( >90%) of resistance determinants, particularly related to β-lactams, suggest that active surveillance of these emergent pathogens is needed in countries like COL. Disclosures Cesar A. Arias, M.D., MSc, Ph.D., FIDSA, Entasis Therapeutics (Grant/Research Support)MeMed Diagnostics (Grant/Research Support)Merk (Grant/Research Support) Lorena Diaz, PhD , Nothing to disclose

A new canine distemper virus lineage identified from red pandas in China.

Canine distemper virus (CDV) is a highly contagious virus that causes multi-systemic, sub-clinical to fatal diseases in a wide range of carnivore species. Based on the sequences of the haemagglutinin (H) gene, CDV strains have been classified into 18 major genetic lineages. In this study, we characterized the genomes of CDV isolated from the lungs of two dead red pandas in China. Histopathological and immunohistochemical analyses revealed damage due to viral infection in these lungs. The two strains showed a deep genetic distance from the other 18 recognized lineages (>4.6% at nucleotide level and>5.0% at amino acid level). The maximum clade credibility tree of the H- gene sequences showed that they belonged to an independent clade and had diverged a relatively long time ago from the Asia-4 lineage (since 1884). These results suggest that the analyzed strains belong to a new CDV lineage, which we designate as Asia-6. Our finding indicates that CDV infections in wildlife in China are complex and are a threat to endangered carnivores.

A morphological, phylogenetic and phylogeographic reappraisal of the land crabs Gecarcinus quadratus De Saussure, 1853, and G. lateralis Fréminville in Guérin, 1832 (Decapoda: Gecarcinidae). Are they different species?

Land crabs of the genus Gecarcinus are distributed along both versants of the American continent. At some point four species were recognized: G. lateralis, G. nobilii, G. quadratus, and G. ruricola. Taxonomically, several authors have tried to differentiate Gecarcinus lateralis from the Atlantic coast and G. quadratus from the Pacific coast. Because morphologic characters used showed high intra-population variability, identification were very difficult. Consequently, these two species have occasionally been considered synonyms. To establish the degree of genetic difference between them a molecular analysis was performed with both species, G. lateralis and G. quadratus, including Mexican Pacific and Atlantic specimens, using the mitochondrial COI and 16S genes. The maximum clade credibility and maximum likelihood trees showed two clades with high support values, one corresponding to the Gulf of Mexico population and the other one to the Pacific coast population, with a genetic distance of 6.1%. The within-clade average distance for the Atlantic populations (G. lateralis) was 1.1% and 0.2% for those of the Pacific coast (G. quadratus). This supports the status of G. quadratus as a valid species, not as a synonym of G. lateralis. The estimated time of divergence between both species ranged between 2.3 to 4.3 million years, coinciding with the proposed closure of the Isthmus of Panama. Morphologically, some populations show high variability in somatic characters, whereas sexual appendices of males of both species showed very little variation. Color in life, shape of sternite 3, and shape and degree of coverage of the abdomen of thoracic sternites, in both sexes, are traits that can be used to differentiate both species.